Oral and Poster Abstracts

130. évfolyam Különszám II. 

Vol. 130. Supplement II. (Oral and Poster Abstracts) 

FÔSZERKESZTÔ – EDITOR-IN-CHIEF 

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Dr. Nagy Béla, Dr. Ózsvári László, Dr. Sályi Gábor, 

Dr. Solti László, Dr. Tanyi János, Dr. Tekes Lajos, 

Dr. Tóth József, Dr. Varga János, Dr. Vetési Ferenc, 

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(Hungarian Veterinary Journal). 

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INDEX: 25531 

Hungarian Veterinary Journal 

Alapította – Established by 

Dr. NÁDASKAY Béla 1878 

HU ISSN 0025-004X

Contents 

Nutrition and Metabolic Disorders 3 

Poster Abstracts 15 

Mastitis / Udder Health and Milk Quality 44 


Infectious and Zoonotic Diseases (Public Health) 74 


Sheep, Goat and Buffaloes 111 

Sheep Poster Abstracts 117 

Goat Poster Abstracts 135 

Buffaloes Poster Abstracts 141 

Camelids 144 


Herd Health Management and Epidemiology 149 


Reproduction and Biotechnology 181 


Lameness 217 


Calf Physiology, Managemenet and Diseases 223 


Animal Behaviour and Welfare 246 


Genetics and Breeding (Crossbreeding) 249 


Good Veterinary Practice (Antibiotic Resistance) 252 


Surgery, Anaesthesia, Pain Management and Diagnostic Imaging 255 


Food Safety 268 


Parasitology 273 


Global Warming 289 


Animal Hygiene, Housing and Biosecurity – Poster Abstracts 291 

Miscellaneous 293 


WORKSHOP 314 

Reproduction Workshop 314 

Perinatal Mortality Workshop 319 

Mycoplasma Workshop 321 

Abortion Workshop 322 

IBR Workshop 323 


Education Workshop 327 


Genetics Diseases Workshop 330 

Contents 1

The World Buiatrics Congress (WBC) has a long tradition. Following the initiative meeting held in Hannover 

(Germany) in 1960, the World Association for Buiatrics was founded at the 2 nd International Congress on 

Cattle Diseases in Vienna in May, 1962. From this time onwards the Biennial World Buiatrics Congresses 

were held in different countries of the world, involving all continents. 

It is a great honour for Hungary to organise the 25 th Jubilee Congress of the World Association for Buiatrics 

particularly because this is the first time the WBC is held in a Middle European country. 

Since the foundation of the world association there have been tremendous developments in the practice of 

bovine medicine and greater emphasis was laid on issues like global control of infectious diseases, metabolic 

disorders caused by malnutrition, herd health management, animal welfare, food safety, environmental 

protection – mentioning only the most important ones. It’s originating from our human nature that we often 

tend to overemphasize the importance of one of the above mentioned areas at the expense of the others, 

forgetting to maintain the healthy balance between them. This attitude appears in the relation of individual 

bovine medicine and bovine herd health management. Our profession requires to treat animals individually 

and to protect their life also on a herd level. We have to find the reasonable balance between individual 

treatments and herd-level preventive measures. We do hope that the 25 th Jubilee Congress will help us to find 

that in our practice. 

The goal of the World Association for Buiatrics is to disseminate scientific and practical knowledge on 

ruminant medicine, management, genetics, production and disease control. The congress will serve as a 

platform for scientists from universities and research institutes, for practitioners working with dairy and beef 

cattle and other ruminants, consultants, residents, post-docs, PhD and graduate students of veterinary 

medicine and animal science in order to provide continuing education for cattle veterinarians worldwide 

every two years. 

We are very proud to mention that this year an outstanding number of 1377 abstracts have been submitted for 

the XXV Jubilee World Buiatrics Congress. Until now, the highest number of abstracts (a bit over 1,000) was 

submitted for the Nice congress in 2006. 

At this year’s Budapest congress 408 of 1,377 abstracts have been selected for oral presentations by our 

international scientific committee. At the same time the majority of the poster presentations (n=860) have 

been accepted by our national organising committee. In cases of poster abstracts there was a very important 

selection criteria namely to give possibilities to the author(s) from all over the world to take part in the 

congress, to experience and practice the mode and level of presentations and to get more familiar with the 

requirements. The oral and poster presentations will be published in the Supplement II of the Hungarian 

Veterinary Journal (Magyar Állatorvosok Lapja) founded by Prof. B. Nádaskay in 1878. 

Notice to Readers 

PREFACE 

Prof. Ottó Szenci Prof. Endre Brydl 

President of the Organising President of the Scientific 

Committee Committee 

Oral and poster abstracts published in the Hungarian Veterinary Journal are not peer-reviewed or referred. All statements, opinions 

and conclusions contained in abstracts in the Hungarian Veterinary Journal are there of the author(s), and are not necessarily those of 

the National and International Scientific Committees.

2 Polyunsatured Fatty Acid in Dairy Cows: 

a Case of Functional Feed (NAT3) 

G. Gabaldo 1 , A. De Palma 3 , A. Fusari 2 , M. Pizzicara 1 , 

S. Tinelli 3 , A. Ubaldi 2 

1 Original Process, Cholet, France 

2 University of Parma, Department of Animal Health, Parma, Italy 

3 Veterinary Practitioner, Verona, Italy 

Omega 3 fatty acids can cross the rumen wall not being saturated by 

saturates. In this mechanism, DHA plays a fundamental rule 

conditioning the rumen flora to convert C18:2 fatty acids in C18:1 

(trans vaccenic acid). This switch in the fatty acids production produces 

an increase in CLA and DHA milk fat fractions other than other types 

of unsaturated fatty acids specific for dairy milk, that ameliorates the 

nutritional characteristics of the milk for humans. 

Experimental study was performed utilising commercial product (FOG3) 

containing an association of olyunsatured Fatty Acid (PUFAs) from 

vegetable-origin like Linolenic Acid and from animal-origin like E.P.A. 

and D.H.A. (NAT), associated with antioxidant molecules (Vitamin E and 

Selenium) and stimulating factors of the rumen fauna (FAP). 

The trial involved totally 44 heads of dairy cows by 5 farms, one in the 

province of Verona (10 treatments and 10 controls), where only the 

zoometric and milk quality indices were tested and 4 farms in the province 

of Bari, where metabolic, endocrinological, fertility, zoometric and milk 

quality indices were tested in 6 cows each farm (3 cases and 3 controls 

each). Biochemical analysis performed stressed that treatments involved 

in this study influenced particularly parameters correlated to lipid status. 

The study confirms that omega 3 fatty acids influenced lipoproteic 

fractions (LDL and HDL) and total cholesterol levels that can be use to 

point out the necessity to administer omega 3 fatty acids. In treated and 

control subjects, plasma progesterone levels were measured. 

Reproduction performance tests showed a good correlation between 

analytical data and the levels of progesterone and the proof of 

pregnancy. In the milk collected by treated cows we noted a significant 

increase in CLA (C18:2) +109,34%, DHA (C22:6) + 223,80%, and 

palmitoleic acid (C16:1) +18.57%, while a decrease in ratios of EPA 

(C20 :5) -57.14%, ALA (C18:3) -20.79% and stearic acid (C18:0) - 

26.09% was observed. 

Key words: PUFAs, omega 3, DHA, CLA, EPA, transition cow, 

fertility 

3 Three Methods of Supplementing Selenium to Intensively 

Grazed Beef Cattle 

J. Maas 

University of California, Veterinary Medicine Extension, Davis, 

United States 

Objectives: To compare the efficacy of supplementing selenium (Se) 

to grazing beef cattle by injection, intra-ruminal bolus, and Se 

fertilization of pastures. 

Materials and Methods: Weaned beef heifers (16 per group) intensively 

grazing irrigated pastures were studied using 4 treatments in 4 replicates 

for 2 years and the trials lasted 120 days. Group 1: one intra-ruminal 

osmotic pump bolus supplying 3 mg Se for 120 days; group 2: 2.5 mg 

Se/45.4 kg subcutaneously; group 3: pastures fertilized with 10g Se per 

hectare (sodium selenate); and, group 4: Se deficient heifers with no Se 

supplements. The Se fertilized pastures were treated prior to the grazing 

season and all other treatments were administered on day 0. Blood 

samples for Se analysis were collected at days 0, 30, 60, 90, and 120 of 

each trial period. Selenium was analyzed by ICP spectroscopy and data 

was analyzed by ANOVA methods. 

Results: Blood Se concentration (ppb; µg/l) data is listed as values for day 

0, 30, 60, 90, and 120, respectively. Year 1. Group 1: 13 ± 4, 94 ± 28, 130 

± 19, 184 ± 15, and 234 ± 27. Group 2: 13 ± 5, 30 ± 6, 20 ± 8, 15 ± 3, and 

8 ± 6. Group 3: 13 ± 5, 113±28, 135 ± 23, 159 ± 20, and 148 ± 16. Group 

4: 12 ± 4, 14 ± 5, 10 ± 6, 5 ± 1, and 4 ± 4. Year 2. Group 1: 16 ± 3, 88 ± 

7, 126 ± 13, 172 ± 19, and 200 ± 19. Group 2: 20 ± 4, 31 ± 4, 27 ± 3, 18 ± 

2, and 10 ± 3. Group 3: 17 ± 3, 46 ± 11, 56 ±9, 46 ± 8, and 40 ± 8. Group 

4: 22 ± 8, 13 ± 6, 9 ± 2, 9 ± 4, and 7 ± 4. The blood Se concentrations were 

significantly (P < 0.0001) increased above controls (group 4) for groups 1 

and 3 in both years and at all times after day 0. Group 2 blood Se 

Nutrition and Metabolic Disorders 

concentrations were increased (P < 0.05) above control cattle only at day 

30 (year 1 and 2) or day 60 (year 2 only). 

Conclusions: The use of intra-ruminal Se boluses or Se fertilization of 

pastures resulted in superior increases in blood Se concentrations in 

cattle intensively grazing irrigated pastures vs. cattle receiving a single 

Se injection. The bolus achieved more consistent increases in blood Se 

concentrations. 

4 Clinicopathological Evaluation of Downer Dairy Cows with 

Fatty Liver 

P. Kalaitzakis 1 , N. Panousis 1 , N. Roubies 2 , N. Giadinis 1 , 

E. Kaldrymidou 3 ,H. Karatzias 1 

1 

School of Veterinary Medicine, Aristotle University, Clinic of Farm 

Animals, Thessaloniki, Greece 

2 

School of Veterinary Medicine, Aristotle University, Laboratory of 

Clinical Diagnosis and Clinical Pathology, Thessaloniki, Greece 

3 

School of Veterinary Medicine, Aristotle University, Laboratory of 

Pathology, Thessaloniki, Greece 

Objective: To evaluate the severity of fatty liver and macro mineral status 

in downer dairy cows and asses the usefulness of determination of serum 

ornithine carbamoyl transferase (OCT) activity, bile acids concentration, 

and other variables for prognosis. 

Materials and Methods: 36 Holstein downer cows were used. Blood and 

liver biopsy specimens were obtained as soon as possible after cows 

became recumbent and before any treatments were administered. Liver 

tissue was examined histologically and classified according to the severity 

of fatty liver. Serum activities of liver-derived enzymes (OCT, GDH, 

SDH and also AST, ALT, GGT, ALP) and concentrations of total lipids, 

triglycerides, bile acids, glucose, ‚-hydroxybutyrate, acetoacetic acid, total 

bilirubin, nonesterified fatty acids, cholesterol and macro minerals (Ca, 

Mg, K, Na, P) were determined. 

Results: The majority of the downer cows had fatty liver infiltration; 

44% were classified as moderate and 44% were classified as severe 

fatty liver infiltration. Survival of the downer cows with moderate and 

severe fatty liver was poor. Serum activities of OCT and GDH, and 

NEFA/cholesterol ratio were indicators of fatty liver. Serum bile acids 

concentration was not an accurate indicator of fatty liver. 

Conclusions: Fatty liver is quite common in downer cows. Serum 

activities of OCT and GDH are useful indicators of fatty liver. 

NEFA/cholesterol ratio is also recommended for diagnosis of fatty 

liver in downer dairy cows, whereas determination of bile acids 

concentration is not. 

Key words: fatty liver, OCT, GDH, downer cows 

5 Vitamin E and Oxidative Damage in Heifers 

R. Bouwstra 1 , R. Goselink 1 , P. Dobbelaar 1 , M. Nielen 1 , 

J. Newbold 2 , T. Werven 1 

1 Faculty of Veterinary Medicine, Utrecht University, Animal Health 

Department, Utrecht, Netherlands 

2 Provimi Research and Innovation Centre, Brussels, Belgium 

The question is whether measurements of oxidative and vitamin E 

status in blood of a periparturient cow are representative for the total 

body, given that blood concentrations of vitamin E and oxidative stress 

products change around this period. This study investigated the 

relationship between oxidative damage and the effect of vitamin E 

supplementation in blood, milk and liver tissue in sixteen periparturient 

heifers. The daily vitamin E intake of the vitamin E supplemented 

Holstein Friesian heifers (n=8) was 3000 international units and started 

two months before calving; the control heifers (n=8) were not 

supplemented. Oxidative damage was determined on the basis of 

malondialdehyde (MDA) concentrations. Blood was sampled nine 

times before calving, on calving day and twice after calving. Liver 

biopsies were taken at week -5, -1, and 2 relative to calving day. Milk 

was obtained from all heifers immediately after calving, the first two 

milkings and on days 3, 7 and 14 at 6 am. Serum and liver tissue were 

analyzed for vitamin E, cholesterol and MDA and milk samples were 

analyzed for vitamin E, MDA, fat, protein and somatic cell count. The 

results show that vitamin E supplements increased both absolute 

vitamin E concentrations and the ratio of vitamin E to cholesterol in 

Nutrition and Metabolic Disorders 3

lood and liver tissue. Absolute vitamin E concentration in milk tended 

to be higher in supplemented cows. Based on the increased MDA blood 

concentrations at calving, it seems that dairy heifers experience 

oxidative stress. The effect of vitamin E on MDA differs between the 

blood, liver and mammary gland. Vitamin E supplementation could not 

prevent the rise in blood MDA at calving, but the significantly lower 

MDA blood concentrations of supplemented cows in the two weeks 

after calving suggest vitamin E has a role in recovery from parturitionrelated 

oxidative stress. Vitamin E supplementation reduced oxidative 

damage in liver, while no obvious effect was found on milk MDA 

concentrations. A strong relationship was found between blood and 

liver tissue vitamin E and the ratio of vitamin E to cholesterol. MDA 

concentrations in blood and milk were also strongly related. The results 

show that the relationship between oxidative damage and vitamin E 

differs within blood, liver tissue and milk. This implies that oxidative 

and vitamin E status calculated on the basis of blood values alone 

should be interpreted with caution and cannot be extrapolated to the 

whole animal. 

Key words: vitamin E, oxidative damage, dairy heifer 

34 Serum Non-Esterified Fatty Acid and Beta-Hydroxybutyrate in 

the Transition Period and their Associations with Disease in 

Dairy Cows 

M. Carson 1 , S. LeBlanc 1 , S. Godden 2 , M. Capel 3 , 

M. Overton 4 , J. Santos 5 , K. Leslie 1 , T. Duffield 1 

1 Ontario Veterinary College, Population Medicine, Guelph, Canada 

2 University of Minnesota, Population Medicine, St. Paul, USA 

3 Perry Veterinary Clinic, Perry, USA 

4 University of Georgia, College of Veterinary Medicine, Athens, USA 

5 University of California, College of Veterinary Medicine, Davis, USA 

Introduction: Serum non-esterified fatty acid (NEFA) and betahydroxy-butyrate 

(BHB) concentrations provide insight into the 

metabolic health of transition dairy cows. The objective of this study 

was to further characterize the relationship of serum NEFA and BHB 

concentrations in the transition period with clinical disease in dairy 

cows across different regions of North America. 

Methods: A field study was conducted using 2403 Holstein cows in 56 

dairy herds in 4 regions of Canada and the United States. Once weekly, 

after the morning feeding, blood was collected from cows in the week 

before their expected calving date, and again from the same cows in 

weeks 1, 2, and 3 postpartum. Serum was stored at -20°C within 8 

hours of collection. NEFA and BHB were measured using a Hitachi 

911 auto-analyzer. 

Results: Results were available from 2149 (confirm) cows. Considered 

alone and assuming equal weight on sensitivity and specificity, the 

optimal cut-points for prediction of LDA were, prepartum: NEFA < 0.5 

mEq/L; week 1 postpartum: NEFA 1.0; or week 1 or 2 postpartum: 

BHB 1400 mol/L (i.e. subclinical ketosis (SCK)). The 23% of cows 

with NEFA 0.5 in week -1 were 2.8 times more likely to subsequently 

have LDA than cows below this cut-point. The 20.5% of cows with 

NEFA > 1.0 in week +1 were 4.6 times more likely to develop LDA. 

The prevalence of SCK and relative risk for affected cows to develop 

LDA were 14% and 4.4 in week +1, and 16% and 3.6 in week +2. Cows 

that did not have elevated NEFA pre- or postpartum or SCK had the 

lowest risk of LDA (0.8%). Conversely, cows that had both high 

postpartum NEFA and SCK, with or without high NEFA prepartum, 

had a 15 to 16% incidence of LDA. The magnitude, timing, and 

duration of elevations in NEFA and BHB combine in a complex 

manner to predict the risk of LDA. Among the 53% of cows with 

NEFA > 0.3 mEq/L in week -1, there was a significant (P = 0.005) but 

modest increase in risk of RP (9% vs. 6%; RR = 1.6). 

Conclusion: These data confirm the associations of NEFA and BHB with 

health and support their use as tools for monitoring or investigation of 

transition dairy cows. However, used alone, the positive predictive value 

of these associations is low, which is expected given the multifactorial 

nature of both RP and LDA. 

35 The Effect of Organic and Inorganic Dietary Selenium Sources 

on Selenium Levels in Blood, Colostrum and Milk and 

Metabolic Profile in Dairy Cows 

D. Kumprechtova 1 , J. Illek 1 , N. Ballet 2 

1 Veterinary and Pharmaceutical University Brno, Clinic of Ruminant 

Diseases, Brno, Czech Republic 

2 Lesaffre Feed Additives, R&D, Lille, France 

4 XXV. Jubilee World Buiatrics Congress 2008 

Objective: The aim was to compare effects of two levels of organic 

selenium (Se) sources and an inorganic selenium source on blood, 

colostrum and milk Se levels, blood chemistry parameters in dairy 

cows under field conditions. 

Material and methods: 36 Holstein cows from 5 weeks (wk) antepartum 

(a.p.) to 12 weeks postpartum (p.p.). Treatments (n=9): 1. NC no 

Se added, 2. PC - 0.3 ppm Se (sodium selenite), S1 0.3 ppm Se (Se 

yeast), 3. S2 - 5 ppm Se (Se yeast). At 5 and 1 wk a.p., at the calving, and 

4, 8, 12 wk p.p., blood samples were analysed for Se, GSH-Px, TP, 

albumin, urea, billirubin, AST, GMT, CK, vit. E. Colostrum and milk 

was collected at the calving and 4, 8 and 12 wk p.p. and analysed for Se. 

Results: Initial Se status in all the cows was marginal. NC showed 

suboptimal Se levels for whole period. S1 and S2 showed significantly 

(P < 0.01) higher values than NC and PC. 

Table 1. Se levels in whole blood 

Treat- NC PC S1 S2 P 

ment (0,3ppm Se) (0,3ppm Se) (5 ppm Se) 

Se (µg/l) Se (µg/l) Se (µg/l) Se (µg/l) 

Mean S.D. Mean S.D. Mean S.D. Mean S.D. 

5 wk a.p. 69,16 5,0 70,3 5,7 77,7 7,3 70,6 6,7 

1 wk a.p. 68,2 a 6,2 90,8 a 7,0 97,7 b 

7,7 155,0 c 22,1 < 0,01 

Calving 67,0 a 5,6 93.6 b 7,9 109,1 c 

7,9 152,2 d 16,5 < 0,01 

4 wk p.p. 65,7 a 5,7 86,1 b 33,7 129,1 c 

7,7 170,8 d 19,8 < 0,01 

8 wk p.p 68,9 a 4,3 105,1 b 7,0 128,6 c 11,2 184,4 d 

9,9 < 0,01 

12 wk p.p. 73,52 a 5,03 110,0 b 15,9 129,22 c 9,58 193,49 d 15,32 < 0,01 

Colostrum Se levels were higher (P < 0.01) in S1 (128.17 µg/l) and S2 

(155.58 µg/l) than in NC (94.68 µg/l) and PC (99.3 µg/l).Overall mean 

milk Se levels in S1 (20.4 µg/l) and S2 (50.0 µg/l) were higher (P < 

0.01) than in NC (10.9 µg/l) and PC (16.6 µg/l). GSH-Px activity in 

whole blood in S1 and S2 was higher than in NC and PC (P < 0.01). CK 

activity decreased with higher org. Se doses. TP, albumin, urea, 

billirubin, AST, GMT, and vit. E were within the physiological ranges, 

non-significant differences. 

Conclusions: Se levels in whole blood, colostrum and milk, and GSH- 

Px activity were significantly increased by organic Se supplementation. 

The response was dose dependent and consistent. Selenium from 

organic source was more bio available than selenium from sodium 

selenite. Dietary Se at a tolerance dose (5 ppm) had no detrimental 

effect on the blood chemistry parameters analysed. 

Project no. 1G46086 (NAZV). 

Key words: dairy cows, organic selenium, blood, milk, metabolic 

profile 

36 The Influence of Subclinical Hypocalcemia on Production and 

Reproduction Parameters in Israeli Dairy Herds 

Ch. Gild, Ni. Alpert, Mi. Van-Straten 

Hachaklait, Ceasaria, Israel 

Introduction: Nearly all mature cows experience some degree of 

Hypocalcemia (HypoC) during the 1st hours post calving. Most cows 

cope with these low Ca concentrations without apparent clinical signs 

however, in some cases Ca concentrations decline to levels that disrupt 

neuromuscular function, resulting in the clinical syndrome known as milk 

fever. It has been established that cows suffering from clinical milk fever 

are susceptible to a variety of secondary problems. To the best of our 

knowledge, no one has evaluated the impact of subclinical HypoC on 

(re)production parameters. Most trials compare the affect of different diets 

on the occurrence of HypoC (clinical and subclinical). Big emphasis is 

given to the subclinical condition although its adverse effects have not 

been proven. Our objective was to investigate the association between 

subclinical HypoC and (re)production parameters in Israeli dairy herds. 

Materials and methods: Blood samples for corrected calcium were 

collected from 634 mature cows from 5 farms. Disease and 

reproduction data were recorded for 180 days. Milk and milk solids 

data originated from monthly milk recordings. Subclinical HypoC was 

defined as serum corrected Ca below 7.5 mg/dl 8-20 hours postpartum 

in the absence of milk fever. Associations between subclinical HypoC 

and retained placenta, metritis, ketosis, stillbirth and uterus prolapse 

were investigated using chi-square tests. Time to conception data was 

analyzed using the Kaplan-Meier method. The relationship between 

subclinical HypoC and milk production was investigated with a 

marginal model for repeated measurements.

Results: Occurrence of clinical and subclinical HypoC was 2.5%, 18.8% 

respectively. Compared with normocalcemic cows, the subclinical 

hypocalcemic cows produced 3.2, 2.7 and 1.9 kg more milk in the first 

three milk recordings, respectively. Sub clinical hypocalcmic cows did 

not have an increased risk for post parturient diseases nor compromised 

reproduction parameters in comparison to normocalcemic cows. 

Conclusions: We could not demonstrate a negative impact of the 

subclinical condition on (re)production parameters. We hypothesize that 

although Ca blood levels decline to sub normal values, the short time 

period and physiological adaptations enable cows to undergo this 

disturbance without apparent (re)production loss. Our results suggest that 

anionic diets to prevent subclinical HypoC are not warranted in Israel as 

the incidence of milk fever is relatively low. 

37 Fatty Liver and Serum Concentrations of Insulin and Glucose 

in Dairy Cows 

D. Kirovsci 1 , H. Samanc 2 , M. Jovanovic 3 , N. Fratric 1 , 

D. Gvozdis 4 , I. Vujanac 2 , B. Dimitrijevic 2 

1 

Faculty of Veterinary Medicine Belgrade, Department of Physiology, 

Belgrade, Serbia 

2 

Faculty of Veterinary Medicine Belgrade, Department of Internal 

Medicine, Belgrade, Serbia 

3 

Faculty of Veterinary Medicine Belgrade, Department of Pathology, 


4 

Faculty of Veterinary Medicine Belgrade, Department of 

Pathophysiology, Belgrade, Serbia 

The aim of this study was to determine the relationship between fatty liver 

and serum concentrations of insulin and glucose in dairy cows during 

peripartal period. Twenty-one Holstein dairy cows were used in this 

study. The cows were classified into three groups (n=7 per group) 

according to the histopathologic findings in hepatocytes: Group 1: cows 

with mild fatty liver, Group 2: cows with moderate fatty liver and Group 

3: cows with severe fatty liver. Liver samples for histopathologic analysis 

were collected from cows by percutaneous needle biopsy on day 10 

postpartum. Liver samples were fixed in paraformaldehyde solution, cut 

and stained with hematoxylin and eosin. Blood serum insulin and glucose 

concentrations were determinated in blood samples obtained from v. 

jugularis at the middle of dry period (30 to 35 days prior parturition), 5 to 

10 days prior parturition and 10 days after parturition. Insulin concentration 

was measured by radioimmunoassay. Glucose concentration 

was measured spectrophotometrically. Insulin concentration was 

significantly lower at day 10 postpartum than 5 to 10 days prior to partus 

(p < 0.05) and 30 to 35 days prior to partus (p < 0.01) in all three groups 

of cows. No significant differences in serum insulin concentrations were 

present among groups of cows in all examined time periods. In Group 3 

serum glucose concentration was significantly lower 10 days postpartum 

than 5 to 10 days prior to parturition (p < 0.001) and 30 to 35 days prior to 

parturition (p < 0.01). In Group 1 significant decrease of glucose 

concentration was determinated from 5 to 10 days prior to parturition to 

10 days postpartum (p < 0.05). There was no significant decrease in serum 

glucose concentration between examinated time periods in Group 2. The 

serum glucose concentration in Group 3 was significantly lower then in 

Groups 1 and 2 both 30 to 35 days prior to parturition (p < 0.05) and 10 

days postpartum. Our results indicate that cows with severe fatty liver 

have low glucose levels, probably because of impaired gluconeogenesis. 

The level of glycemia was not correlated with serum insulin 

concentrations which indicate that glucose concentration is under 

complex endocrine control during peripartal period. 

Key words: dairy cows, fatty liver, insulin, glucose 

38 Metabolic Syndrome in Cows: TNF-1 and Fat Metabolism in 

Heifers with Different Body Condition (BCS) at Parturition 

M. Fürll, N. Jatzke, K. Bauer, I. Steinhöfel, J. Gottschalk, 

U. Sack, A. Einspanier 

University of Leipzig, Internal Medicine, Leipzig, Germany 

Objectives: Recent studies in humans and other animals have shown 

that fat tissue is not a simple energy storage organ, but releases a lot 

of molecules which exerts important endocrine and immune 

functions. Adipocytes secrete numerous bioactive substances, 

including hormones, growth factors and cytokines. Abdominal fat 

accumulation has been shown to play crucial roles in the 

development of the human metabolic syndrome. This study 

describes the effect of body condition on metabolic profiles, 

hormones and cytokines in Holstein cows during the peripartal 

period. Maybe there are some similarities with the components of the 

human metabolic syndrome. 

Material and methods: 15 cattle were fed with a high energetic diet 

(hED) until they became pregnant, 15 heifers were chosen because 

of their high back fat thickness (hBFTh), and 15 cattle were fed in 

optimum conditions (oED). Blood samples were taken 1 week before 

parturition (w a.p.), 3 days after parturition (d p.p.) and 1 month p.p. 

to determine metabolic, hormonal as well as immunological 

parameters. 

Results: The FFA, insulin and TNF1 concentration (medians) are 

shown in the table. 

FFA (mmol/l) insulin (nmol/l) TNF 1 (ng/ml) 

1 w 3 d 3–5w 1 w 3 d 3–5 w 1 w 3 d 3–5 w 

a.p. p.p. p.p. a.p. p.p. p.p. a.p. p.p. p.p. 

hED 160 610 200 0,16 0,07 0,12 27 33 29 

oED 190 450 275 0,13 0,09 0,09 10 22 10 

hBFTh 270 612 230 0,16 0,16 0,13 10 10 34 

In high back fat thickness, the cows insulin concentrations were 

significant higher at the third d p.p. than in the other groups; the high 

back thick fatness cows showed the highest glucose concentrations 

during the whole sample period. We found a positive correlation 

between TNF-1 and FFA concentrations in cattle with hBFTh 

(r=0.63). In hBFTh heifers and in cattle the concentrations of IGF-I 

were significant lower at day 3 p.p. Not only a positive correlation 

existed among the IGF-I and the FFA concentrations (r=-0.52) but 

also a negative correlation among IGF-I and haptoglobin in all 

groups (r=-0.61). 

Conclusion: In conclusion the study shows that changes of energy fat 

metabolism - including TNF-1 from adipose tissue is comparable with 

the metabolic syndrome in humans and may be directly involved in the 

pathogenesis of many postpartum disturbances. 

Key words: metabolism, body condition, metabolic syndrome, heifers 

39 Influence of Gluconeogenesis on Metabolic Status and 

Appereance of Oxydative Stress in Peripartal Cows 

B. Dimitrijevic 1 , S. Borozan 2 

1 

Faculty of Veterinary Medicine Belgrade, Clinic for Ruminants and 

Pigs, Belgrade, Serbia 

2 

Faculty of Veterinary Medicine Belgrade, Chemistry, Belgrade, Serbia 

In ketotic condition of cows, due to presence of ketone bodies, 

appearance of reactive oxygen species (ROS) and reactive nitrogen 

species (RNS) might occur. Durable presence of ketone bodies may 

cause cells susceptible to irreversible damage, what is reflected in 

increased lipid peroxidation through damaging of cell membranes, as 

well as increased level of lactate dehydrogenase (LDH) enzyme in 

extra cellular space. The objective of this study was to examine impact 

of keton bodies on ROS and RNS production, levels of lipid and 

protein damage in different level concentration of keton bodies in 

sample of cows blood. Research was conducted on sample of Holstein 

Friesian cows (n=15), during few first days after calving, and presence 

of keton bodies was proven. Control group was made up of cows with 

negative test on ketone bodies. Production of ROS was examined 

through activity of superoxide dismutase (SOD) and catalase (CAT) 

enzymes in erythrocytes, whereas production of RNS was monitored 

through the contents of nitrites in blood plasma of the examined 

animals with ELISA. The level of lipid peroxydation was monitored 

through malondialdehyde (MDA). The effect of free radicals provoked 

by keton bodies on hepatocytes was monitored through determination 

of izoenzymes distribution of LDH (LDH1-LDH5) by electroforesis. 

Research results showed significantly increase in SOD activity 

(P

RNS causes increased lipid peroxidation and accelerated proteolysis 

what is reflected in increased presence of LDH5, as hepatocytes 

destruction. 

Key words: cow, keton bodies, oxydative stress, ROS, RNS 

70 Effect of BCS Change during the Dry Period on the 

Performance of Dairy Cows 

T. Van Werven, D. Speksnijder, J. Papen, M. Nielen 

Faculty of Veterinary Medicine, Utrecht University, Farm Animal 

Health, Utrecht, Netherlands 

This abstract shows the preliminary results of an on going blind trial 

on the effect of orally supplemented vitamin E during the dry period 

on the performance of dairy cows. Most studies are interested in 

either calving BCS or amount of BCS loss postpartum. This study 

evaluates the effect of BCS change (loss or gain) during the dry 

period on the postpartum performance of the cow. Five dairy farms 

were selected to participate in a blind field trial to investigate the 

effect of orally supplemented vit E. From each farm approximately 60 

dry cows are enrolled into the study 6-8 weeks prior to calving date. 

Dry cows are divided in two groups (A and B), one group is fed 135 

IU and the other group 3000 IU of vit E during the dry period. Body 

condition score is measured at a 5-point scale once a week during the 

dry period, at calving and at the first milk test-day. Milk test-day 

records are collected for fat, protein and SCC. Data on retained 

placenta are collected and defined as failure to expulse the placenta 

within 12 hours pp. Change of body condition score during the dry 

period is calculated by regression analysis which provides cows with 

an increasing or decreasing BCS during the dry period. Cows with an 

unchanged BCS during the dry period are included in the group of 

cows with increased BCS. Until so far milk test-day records of 94 

cows are presented. At the beginning of next March all cows will be 

included. From these preliminary results some interesting trends are 

demonstrated. There seems to be a difference between the two 

vitamin E groups (blind). Cows in group A show more often an 

significant increase of BCS during the dry period than cows in groups 

B, corrected for herd effects. Cows with average BCS3, corrected for increasing or decreasing slope during the dry 

period. Preliminary trends indicate that cows with a BCS loss during 

the dry period are less at risk for a fat/protein ratio > 1.4 than cows 

that gained BCS, corrected for days in milk. These results indicate 

that cows with BCS loss in the dry period might have a more efficient 

energy metabolism after parturition and therefore be less at risk for 

metabolic diseases like ketosis after parturition. Those cows might 

experience fewer metabolic changes during the transition from late 

gestation to early lactation. The possible effect of vit E on BCS needs 

further research. 

Key words: body condition score, dry period, dairy cows, vitamin E 

71 Metabolic Syndrome in Cows: TNF-1 Concentrations before 

and after Parturition in Healthy and Ill Cows 

B. Fürll 1 , G. Hädriich 2 , M. Fuerll 2 

1 

University of Leipzig, Department of Veterinary Physiology, Leipzig, 

Germany 

2 

University of Leipzig, Department of Internal Medicine, Leipzig, 

Germany 

Objectives: Today, cattle health is especially disturbed because of a 

strained metabolism. Cytokines of the visceral fat depot, TNF-1 for 

example, can affect the metabolism negatively. Therefore we 

investigated cows with the aims: a) Are there any differences 

between the TNF-1 concentrations in healthy and ill cows around 

parturition? b) Can those differences be noted before parturition 

already? c) Are there any TNF-1 changes specific for certain 

diseases? 

Material and methods: 20 healthy cows as well 103 cows with 

various diseases (table) were examined 10 days before parturition (d 

a.p.) and 3 d after parturition (p.p.). Blood samples were analysed 

byHitachi 912, TNF-1 by an ELISA. 

Results: The TNF-1 concentrations a.p. and p.p. of healthy (medians 

36:37 ng/ml) and ill (140:55 ng/ml) cows differed significantly. Table: 

TNF-1 (ng/ml), FFA (mmol/l) and haptoglobin (Hp, g/l) concentrations 

(medians) in healthy cows and cows with different diseases p.p. 

(*p=0.05) 


d p.p heal- ret. endo- still- ova- twin mas- Dislothy 

pla- met- birth rian birth titis catio 

centa ritis cysts abomasi 

(DA) 

TNF-1 -10 35,6 90,7* 50,4 26,8 23,9 33,4 76,4 55,9* 

3 37,3 117,5* 47,8 19,9 10,0 19,8 50,5 10 

FFA -10 80 115 180* 150* 115 310 210* 85 

3 570 870* 872* 790* 875* 875* 620 1270* 

Hp -10 0.20 0.09 0.33* 0.09 0.11 0.40* 0,08 0,28 

3 0.74 1.51* 1.52* 1.45* 1.22* 1.86* 1,01 1,04 

The TNF-1 concentrations correlate a.p. with the back fat thickness 

(p>0.05); they are at their highest (p 0.4 mmol/L, and the prevalence of herds with >15% 

subclinical ketosis (cut-off of 1.4 mmol/L) was 52.6%. The within herd 

prevalence of NEB and SCK ranged from 14% to 80% and 8% to 44%, 

respectively. The prevalence of SARA (rumen pH 15% elevated NEFA and in 75% of 

herds with >15% elevated BHB. At least one retained placenta was 

reported in 85% of herds with >15% elevated NEFA and in 100% of 

herds with >15% elevated BHB. Further modelling of the data will be 

discussed at the meeting. The results suggest that subclinical metabolic 

disease in transition cows is present in dairy herds surrounding Lima, 

Peru at levels that may negatively impact milk production, 

reproductive performance, and profitability. There is opportunity to

improve management practices to minimize subclinical disease in 

transition cows and maximize profitability. 

Key words: metabolic disease, transition cows, subclinical ketosis, 

negative energy balance, Lima Peru 

73 Evaluation of three Laboratory Methods Measuring NEFA in 

Cow’s Blood 

T. Gaal, P. Ribiczey 

Szent István University Faculty of Veterinary Science, Department of 

Internal Medicine, Budapest, Hungary 

Non esterified fatty acids (NEFA) in the blood are good markers of 

negative energy balance (NEB) in dairy cows. There are several 

laboratory methods to assess NEFA concentration. One method is 

based on a colorimetric, non-enzymatic technique at room temperature 

(Duncombe, 1964), the other one applies colorimetric, enzymatic 

determination at 37 °C (Matsubara et al., 1983). Recently a portable 

photometer appeared on the market (DVM-NEFA ® ) using a 

colorimetric technique for NEFA measurement at room temperature. 

As laboratory methods need a well-equipped laboratory with skilled 

personnel, introduction of a cow-side test is more than welcome. 

However, widely used NEFA reference values were established by the 

laboratory methods, therefore critical evaluation of DVM-NEFA is 

recommended. The goal of this study was to compare NEFA results 

determined with 3 methods. On a herd of 900 HF dairy cows blood 

samples were collected from dry cows (n=17) and fresh cows (n= 14). 

Blood serum NEFA was measured immediately by 3 methods. For the 

non-enzymatic method commercial chemicals were used while for the 

enzymatic method a Randox NEFA test kit was applied on an 

automatic chemistry analyzer. Results were analyzed by t-test and 

regression analysis. Pre-partum NEFA values determined by DVM- 

NEFA, non-enzymatic and enzymatic methods were 0.258+0.200, 

0.105+0.090 and 0.093+0.088 mmol/L, respectively. Post-partum 

these were 0.706+0.327, 0.300+0.146 and 0.293+0.179 mmol/L. Prepartum 

NEFA was lower than post-partum with all methods (P

An experiment was conducted to study the influence of calcium 

soaps (CAS) or triglyceride (HTG) of hydrogenated palm oil on liver 

and blood parameters indicating the energy status and lipid 

metabolism of cows in the peripartum period. Holstein-Friesian 

dairy cows were fed on a corn-silage based diet supplemented with 

11.75 MJ NEL per day of CAS or HTG or without fat 

supplementation (control) in a period from d 21+3 prior to the 

expected calving to d 100+5 postpartum. At d 21+2 (basal sample) 

and d 15+2 prepartum and d 5+2, 25+3 postpartum liver biopsy and 

blood samples and at d 100+5 postpartum an additional blood 

sample were collected from the cows. Total lipid (TL), glycogen 

(GL) content, fatty acid composition of liver lipids and glucose 

(GLU), insulin (INS), free fatty acid (NEFA), beta hydroxy butirate 

(BHBA), triglyceride (TG), cholesterol (Ch), aspartat-aminotransferase 

(AST) contents from the blood were determined. No 

differences were detected in the liver fat among the groups at d 14 

prepartum or d 25 postpartum. At d 5 postpartum, however, both 

control and CAS-cows had higher TL and lower GL concentrations 

in the liver than HTG-cows (P

only for less than 3 days. All treated wet corn samples (both varieties) 

protected their stability for 7 days. The fermentation dynamic, aerobic 

stability, mould and mycotoxin examinations have proven that from 

feed hygienic point of view this technology is more advantageous 

comparing to the traditional technologies. Taking into consideration 

the technology and the feeding requirements this technology can be 

fully inserted to the cattle feeding system. 

Key words: corn, crimping, preservation, fermentation, mycotoxin 

160 Necessity of the New Biotechnological Methods for Avoiding 

Subacute Ruminal Acidosis 

S. Latos 

Dr. BATA Biotechnological Co, Reserch & Development, Ócsa, 

Hungary 

There are several strategies for feeding dairy cows, but all of them based 

on the roughages, rich in fiber. The ingredients added to the ration 

(energy-, and protein carriers, vitamins, minerals) can be used for filling 

the gap between the value of the basic forages and the requirements of the 

cow. To fulfill the energy demand of the early lactation, we have to 

increase the energy and decrease the fiber content of the ration. The most 

used energy sources are the treated cereals. The treatments are increasing 

the digestibility or the rumen metabolisation rate of the non-structural 

carbohydrates in the cereals.The high yielding status can only be 

maintained for short time periods but the larger problem is, that these 

rations are harmful for the health status of the cow. In these cases evolves 

the subacute ruminal acidosis (SARA). It can be defined as a temporary 

rumen state, with moderate pH decrease (pH 5,5-5,0) in the rumen for 

several hours. Cows suffering in SARA are showing the symptoms such 

as decrease in dry matter intake and in yield, lower milk fat, diarrhea, body 

condition loss. Not only a ration with high level of NSC can cause SARA, 

management faults (feeding techniques, sludge feeding, selection) can 

support the development of the problem. Even the environment (heat 

stress) can play a role to SARA.The fermented cereals (wet corn) taking 

more place in the last years can also higher the risk of SARA. They 

contain high amount of lactic acid and living Lactobacilli. In the acidotic 

rumen (anaerobic environment, pH>5,5) they can activate and start to 

produce even more lactic acid which can lead to an acute ruminal 

acidosis.In practical farm circumstances, with high NSC and critical NDF 

level rations it is almost impossible to avoid the SARA. This shows the 

subservience of using biotechnological products and methods which can 

help to avoid or minimize these damaging processes due to their 

biological activity. 

A typical example the world-wide used live yeast (Saccharomyces 

cerevisiae), which can directly affect the fermentation, unlike the so called 

yeast-metabolites. It is able to consume the oxygen in the rumen, and 

reacts with the lactic acid utilizing microorganisms. This helps to stabilize 

the pH, consequently minimizes the risk of SARA, resulting not only a 

higher production but furthermore a better health status and fertility rate. 

Key words: rumen acidosis, biotechnology, prevention 

161 Comparison of the Effect of Feeding of Crimped versus Dry 

Corn Grain in High Producing Dairy Cows 

E. Brydl 1 , A. Tirián 1 , L. Könyves 1 , V. Jurkovich 1 , L. Tegzes 1 , 

R. Farkas 1 , L. Horváth 2 , M. Török 2 , J. Karnóth 3 , H. Miettinen 4 

1 Szent István University Faculty of Veterinary Science, Department 

of Animal Hygiene, Herd Health and Veterinary Ethology, 

Budapest, Hungary 

2 Gorzsa Agricultural Co, Dairy Farm, Hódmez_vásárhely, Hungary 

3 NOACK Hungary Ltd, R&D, Budapest, Hungary 

4 KEMIRA OYJ, R&D, Helsinki, Finland 

The aim of the study was to measure the effect of feeding of crimped corn 

grain with high moisture (DM 70%) vs. dry one on the rumen 

fermentation, energy, protein and acid-base metabolism, health status, 

reproductive performance, daily milk production and milk composition 

of high producing dairy cows. Kemisile 2S was used as preservative at 

crimping with 4.2 kg/t. The experiment was carried out at a dairy farm 

with 1000 head of Holstein Friesian cows. Two hundred in calf second 

and third parity Holstein Friesian cows were selected randomly to 

control and experimental groups, 3 weeks prior to expected parturition. 

Rumen fluid, blood and urine samples were taken for laboratory 

examinations once a week in the first month and once a month onwards 

the experiment for four times. The corn silage based diet was identical for 

both groups except for the control animals received dry and the 

experimental cows crimped corn. The total concentration of volatile fatty 

acids in rumen fluid of the experimental cows was superior to the 

controls by 12.1% on average. The acetate ratio of the experimental 

cows was superior to the controls by 13.4% on average (p

2 

Hungarian Institute of Agricultural Engineering, R&D, Gödöllô, 

Hungary 

of study: The authors investigated a new bale-forming system: dry 

matter limitation of the new technology and effect of short (29% 

DM: 4 hour) and long term wilting (52% DM: 12 hour) on bale 

parameters and fermentation profile in new-type alfalfa bales. 

Materials and methods: Alfalfa was mowed with theoretical chop 

size of 20-30 mm. Baling was carried out by a Göweil LT Master 

variable chamber baler-wrapper machine. Pressurization: 150 bar. 

Output: 18-20 bales hour -1 for alfalfa. Film for wrapping was applied 

with thickness of 25 µm in 7 layers, respectively. The pH, lactic and 

volatile fatty acid composition were analysed on the 13 th , 30 th and 70 th 

day of fermentation according to the Hungarian National Standards 

(Hungarian Feed Codex, 2004). 

Results: Extremely high density was carried out with the new 

technology: 213-278 DM kg m -3 due to high pressurization (150 bar) 

and small particle size (20-30 mm) compared to conventional bales 

(90-200 DM kg m -3 ) (Table 1). Better homogeneity of the new-type 

bales compared to the conventional bales, owing to chopped and 

mixed forage, was confirmed by the low variation in wet bale weight 

(var. coeff. 1.4%). High density results extremely good anaerobe 

circumstances for fermentation (Table 2). It was found higher pH, 

lactic acid content and lower acetic acid content in bales with 52% 

DM-content. 

Table 1. Bale characteristics in alfalfa baled with the new baler system 

(n=15) 

29% DM 52 % DM 

Bale wet weight (kg) 904±25,1 a 657±13,0 b 

Bale dry weight (kg DM) 262±7,3 a 342±6,6 b 

Wet density (kg m -3 ) 734±10,3 a 534±7,9 b 

Dry density (kg m -3 ) 213±3,0 a 278±4,1 b 

Table 2. Fermentation characteristics in alfalfa baled with the new 

baler system (n=5) 

Day of Treat- pH Total Lactic Acetic Butyric 

fermen- ment acid 

g kg 

acid acid acid 

-1 

g kg -1 

g kg -1 

g kg -1 

13 th day 29% DM 4,84 a 28,9 a 21,3 a 7,6 a 0,0 a 

52 % DM 4,87 a 30,0 a 24,4 a 5,5 b 0,0 a 

30 th day 29% DM 4,60 a 34,8 a 26,9 a 7,8 a 0,0 a 

52 % DM 4,79 b 37,6 b 30,8b 6,7 a 0,0 a 

70 th day 29% DM 4,49 a 32,9 a 25,2 a 7,6 a 0,2 b 

52 % DM 4,74 b 35,1 a 28,2v 6,8 a 0,1 a 

Conclusions: It was confirmed that the new baling system is able to 

form bales in a wide range of dry matter content (290-520 g kg-1 in 

alfalfa) with high density and excellent fermentation profile. 

Key words: bale-forming technology, high density, fermentation 

quality, alfalfa silage 

164 Periparturient Risk Assessments for Placenta Retention in 

Dairy Cows 

L. Könyves 1 , O. Szenci 2 , V. Jurkovich 1 , L. Mrs. Tegzes 1 , 

A. Tirián 1 , N. Solymosi 3 , Gy. Gyulay 4 , E. Brydl 1 


of Animal Hygiene, Herd Health and Veterinary Ethology, 


2 Szent István University Faculty of Veterinary Science, Clinic for 

Large Animals, Budapest, Hungary 


of Biomathematics, Budapest, Hungary 

4 Private practitioner, Martonvásár, Hungary 

The aim of the study: was to measure how the metabolic parameters 

of energy and acid-base metabolism indicate the risk of placenta 

retention (PR) in dairy cows. The relationship between animal- and 

environment related factors and PR, the consequences of PR for 

subsequent puerperal endometritis (PM), metabolic status, milk 

production and reproduction were analysed as well. 


Materials and methods: 105 clinically healthy Holstein Frisean cows 

were assigned randomly at 21-14 days before expected calving. Blood and 

urine samples were taken for metabolic profile test on

1 

Szent István University Faculty of Veterinary Science, Department 

of Animal Hygene, Herd-health and Veterinary Ethology, Budapest, 

Hungary 

2 

Gorzsa Agricultural Co, Dairy Farm, Hódmezôvásárhely, Hungary 

3 

Dr. BATA Biotechnological Co, Research & Development, Ócsa, 

Hungary 

The aim of the study was to measure the effect of feeding of micro 

capsulated viable yeast on the rumen fermentation, health and milk 

production of diary cows. The experiment was carried out at a large scale 

dairy farm in Hungary between February and August 2007. The milk 

production of the cows was nearly 10 000 l yearly. Two hundred in calf 

second and third parity Holstein Friesian cows were selected randomly to 

control (No Yeast Received, NYR) and experimental (Micro capsulated 

Viable Yeast) groups, 3 weeks prior to expected parturition. Rumen fluid, 

blood and urine samples were taken for laboratory examinations once a 

week in the first month and once month onwards the experiment six times. 

The total concentration of volatile fatty acids (VFAs) was normal in both 

groups however in the experimental cows it was higher by 4.4% and the 

rumen fluid pH was more stabile. Lower concentration of NEFA, BHB 

and urea concentration was found in the experimental cows one month 

after parturition indicating more balanced energy metabolism and better 

health. The average daily milk production was higher by 0.6 l and the days 

spent for different diseases e.g. mastitis, lameness etc. was less by 33 d in 

the experimental group. The reproductive indices were more favourable in 

the experimental animals than in the controls. This work was supported by 

the project of GAK ALAP1-00003/204. 

Key words: micro capsulated, viable yeast, dairy cow, rumen 

fermentation, health, milk production 

233 Subacute Rumen Acidosis in Intensive Italian Dairy Herds: 

Occurrence and Clinical Approach 

M. Gianesella 1 , C. Stelletta 1 , C. Cannizzo 1 , 

P. De Battisti 2 , M. Morgante 1 

1 Universita’ Degli Studi di Padova, Dipartimento di Scienze Cliniche 

Veterinarie, Legnaro (Pd), Italy 

2 Cortal Extrasoy S.P.A., Cittadella (Pd), Italy 

Subacute rumen acidosis (SARA) represents one of the most 

important metabolic disorders in intensive dairy farms that affects 

rumen fermentations, animal welfare, productivity and profitability. 

The aim of the present study was to study the occurrence of SARA in 

intensive Italian dairy herds and to determine the correct clinical 

approach to evaluat this disease. During the last four years forty 

commercial dairy herds were investigated. In each farm a general 

investigation was performed, with particular attention to herd 

management and presence of typical clinical signs of SARA. Twelve 

cows in good body condition, between 5 and 60 day in milk and 

without clinical signs of disease were selected randomly from each 

herd, to perform rumenocentesis and obtain rumen fluid. Ruminal pH 

was determined immediately after sampling and concentration of 

SCFA in ruminal fluid was determined on samples after storage. 

Comparisons between TMR after distribution and residual food were 

be evaluated by sieving and by chemical composition. Milk yield 

quality was determined in the cows were we performed the 

rumenocentesis. Results were subject to ANOVA and correlation 

analysis using SIGMA STAT 2.03. We had no sampling problems 

with the 480 cows of our study series, and no animal developed any 

health problem during or after the procedure. The results indicated the 

presence of SARA in 40% of herds (more than 33% cows with rumen 

pH < 5.5), a critical situation (more than 33% cows with rumen pH < 

5.8) in 25.7% of farms and a normal rumen pH condition in 34.3% of 

herds. In particular dairy herds show an average SCFA concentration 

of 162, 147 and 121 mmol/L for low pH, critical pH and normal pH 

herds, respectively. The differences among diet composition were not 

significant even if herds with SARA showed a light discordance 

between initial composition and residual feed, a result which suggests 

more attention in TMR preparation and management, especially in 

large herds where the attitude of the personnel responsible for feeding 

the cows is of paramount importance. Milk yield quality were not 

statistical different in three groups: in particular dairy herds show an 

average of 3.8, 3.6 and 3.5 of milk-fat % for low pH, critical pH and 

normal pH herds, respectively, and the same values (3.3) of milk 

protein %. Although ruminal pH is reported to drop in experimental 

situations of inducing low milk-fat, this alone does not justify those 

situations to be characterized as being SARA. 

234 Measuring Rumen pH and Temperature by an Indwelling 

and Wireless Data Transmitting Unit – Application under 

Different Feeding Conditions 

J. Gasteiner 1 , S. Rosenkranz 2 , M. Fallast 2 , T. Guggenberger 1 

1 Federal Research Institute Raumberg-Gumpenstein, Animal Care 

and Animal Health, Irdning, A-8952, Austria 

2 University of Technology Graz, Sciencepark, Graz, Austria 

Subacute rumen acidosis is a common and economically important 

herd health problem of dairy cattle and there is a crucial need for 

monitoring systems. 

An indwelling wireless data transfer system for monitoring rumen ph 

and temperature was assembled. Readings (user selectable) were taken 

every 30 minutes. Stored data are transmitted using a radio transceiver 

communicating with the ISM-band frequency 433 MHz. The system is 

controlled by a microprocessor. Data (pH, temperature) are sampled 

with an A/D converter and stored in an external memory chip. The 

latest class of the indwelling system can be administered orally, but to 

service the measuring units, experiments were conducted using 5 

ruminally cannulated steers. 

After calibrating and check for proper operation rumen pH und 

temperature measures were carried out under following feeding 

conditions: 1. 100 % roughage for 1 week, measuring last 3 days 2. 

Daily pasture and roughage ad lib. for 3 weeks, measuring last 7 days 

3. 50:50 roughage: concentrate for 7 days, measuring 7 days 

In experiment 3, 2 measuring units were placed simultaneous in one 

animal to compare and to verify data. At the same time, rumen samples 

were taken at intervals (1hour) and pH was detected with a pH meter. 

Statistical analysis was conducted using GLM (Statgraphic Plus 5.1) and 

Bonferroni-Holm-Test. 

Experiment 1: rumen temperature (mean 38.40±0.70 °C) was influenced 

significantly by drinking water but it is not connected with feeding time. 

Mean pH was 6.49±0.39 and nadir was pH 6,14. Experiment 2: mean 

rumen temperature was 38.12±0.80 °C and mean pH was 6.36±0.22. 

Nadir during pasture (day) was pH 5.34, nadir during feeding roughage 

(night) was 6.16. Pasture had a significant influence on rumen pH. 

Experiment 3: mean rumen temperature was 38.55±0.83 °C and mean pH 

was 6.37±0.24. Nadir was pH 5.29. Decline of rumen pH was 

significantly related to the feeding of concentrate. When comparing the 

results of 2 simultaneous measuring units, the absolute statistical error 

was 0.6±0.65 °C for temperature and 0.15±0.19 for pH, differences could 

be seen due to rumen dynamics and non-homogenous mixture of ingesta. 

Results indicate that the present method is a useful tool for scientific 

applications. As the measuring system can also be administered to 

uninjured cattle, an adapted rumen pH measuring system will also be 

assembled for practical purposes in future. Examinations were carried 

out in compliance with the National Animal Experiment Law 

(Tierversuchsgesetz 1988) 

235 Blood and Ruminal Biochemical Modifications and Clinical 

Signs after 1 and 2 % Body Weight Intoxication with Soybean 

Meal in Dairy Steers 

D. Raboisson, A. Ferrieres, F. Enjalbert, F. Schelcher 

National Veterinary School of Toulouse, Department for Farm 

Animals and Herd Health, Toulouse, France 

Effects of massive ingestion of rapidly fermentable carbohydrate diets on 

cattle are well known, but no data are available on massive ingestion of 

soybean meal. The aim of the trial is to describe the clinical signs and the 

ruminal and blood biochemical modifications of bulls receiving an large 

quantity of soybean meal. In two successive trials (8 weeks apart), 

soybean meal (46% crude protein) was introduced via the canula in the 

rumen of four Holstein steers, respectively at 1% (3,5 to 3,7 kg) and 2% 

body weight (7,8 to 8,3 kg). Clinical examination, ruminal and blood 

biochemical analysis were performed every two hours for 4 days. 

Measured parameters were blood pH, PCO 2 and HCO 3 - (Opti-CCA, 

Idex), blood NH3 (Vitros 250, Ortho Clinical Disgnostic), ruminal pH 

(pH-meter), ruminal volatile fatty acids (VFA) (gas chromatography) 

and ruminal N-NH3 (spectrophotometry). In the 2% study, ruminal pH 

firstly decreased to 5,67 ± 0,15 at H 4 (4 hours after soybean meal 

ingestion), in parallel to an increase of VFA concentrations and a 

moderate and stable concentration of N-NH 3 between H 4 and H 12. Then 

ruminal pH increased slowly to basal level (pH= 6,47 ± 0,18) up to H 20 

(VFA decreased and N-NH 3 increased from H 12 to H 26). Ruminal pH 

finally increased slightly up to H 32 and returned to basal level. Blood pH 


and plasma HCO 3 - increased from H4 to H 40 and then decrease to basal 

level until H 80. Blood NH 3 concentration peak (H 20 : 378,2 ± 103,3 

µmol/L) was detected a few hours after blood pH peak (H 12). Uremia 

peaked at H 26. Hyperammoniemia was associated with an hyperglycemia 

(H 24). Clinical signs (moderate muscle tremors, head shaking, 

stamping, weackness and moderate prostration) occurred in the 2% trial 

between 12 and 28 hours after soybean meal ingestion. Clinical signs 

were detected during hyperamoniemia, suggesting causual relationship 

between symptoms and blood NH 3 concentration. Similar but less 

demonstrative biochemical modifications occured with 1% soybean meal 

ingestion, but no clinical sign was detected. Clinical signs and major 

biochemical modifications occured only with ingestion of large 

quantities of soybean meal, but spontaneous ingestion of such a quantity 

by cows is unlikely. 

236 The Effect of Rumen Mucosal Lesions on Growth in South 

African Feedlot Cattle 

P. Thompson 1 , W. Schultheiss 2 , A. Hentzen 3 

1 

University of Pretoria, Production Animal Studies, Onderstepoort, 

South Africa 

2 

Schering-Plough Animal Health, Johannesburg, South Africa 

3 

Supreme Livestock Services, Heidelberg, South Africa 

Rumen health is well recognized as a primary necessity for bovine 

health and if compromised it will negatively affect performance. It is at 

risk when abrupt transition to a more energy dense ration takes place 

early on in the beef feedlot feeding period. This is also the time when 

most cases of undifferentiated bovine respiratory disease (UBRD) 

occur. Excessive pH fluctuations may cause subclinical or clinical 

acidosis and result in permanent damage to the rumen mucosa. 

Although such lesions are commonly seen in apparently healthy cattle 

at slaughter, there are no reports of their effect on the performance of 

feedlot cattle. The rumen mucosa of 1935 cattle was examined at slaughter, 

representing 15 groups from five feedlots. The presence and size of focal 

mucosal scarring (“stars”) and diffuse mucosal lesions were recorded. Lung 

lesion scoring was also done on most animals. Live mass on arrival at the 

feedlot and average daily gain (ADG) during the feeding period were 

obtained for each animal. The effects of rumen mucosal lesions on ADG 

were then estimated using mixed-effects multiple regression models. 

Stars were recorded in 50.9% (varied from 3.3% to 72.3% between groups) 

and diffuse lesions in 56.1% (5.5% to 93.6%) of all animals. Pulmonary 

lesions were seen in 52.3% (32.3% to 77.1%) and severe pneumonia (lung 

parenchymal or pleural adhesion score >2) in 13,6% (4.8% to 33.9%) of all 

animals. Mean ADG was 1.406 kg (1.176 to 2.070 kg). When adjusted for 

processing mass, sex, feedlot and group, calves with rumen stars showed an 

average reduction in ADG of 46 g (P < 0.001). The presence of a diffuse 

mucosal lesion was associated with a 60 g reduction in ADG (P < 0.001). 

Calves with both lesion types showed a 79 g reduction in ADG (P < 0.001). 

However, the effect of each of the rumen lesions on ADG varied between 

groups. In one group of 185 animals, the presence of rumen stars was 

positively associated with the presence of severe pneumonia (odds ratio = 

4.9; P = 0.03). This information is important for feedlot managers, 

supporting the fact that transition from adaptation to grower or finisher 

rations should be made with care. Efforts should be made to limit pH 

fluctuations which may cause permanent damage to the rumen mucosa 

that will increase days on feed to attain the desired end mass. Routine 

inspection of rumen mucosa at the abattoir may be also considered as a 

tool to monitor nutritional management in the feedlot. 

Key words: rumen mucosal lesions, pneumonia, feedlot cattle, growth 

237 Case Report of Iodine Deficiency in an Organic Dairy Farm 

I. Schlemmer 1 , M. Metzner 1 , S. Reese 2 , U. Ebert 3 , W. Klee 1 

1 

University Munich, Clinic for Ruminants, Munich, Germany 

2 

University of Munich, Institute of Veterinary Anatomy, Munich, 

Germany 

3 

Bavarian Health and Food Safety Authority, veterinary pathology, 

Munich, Germany 

In an organic dairy farm with 40 Holstein-Friesian cows and offspring 

in Bavaria the farmer described tumours in the region near the larynx 

in five of 15 calves (up to the age of six months). Additionally the 

growth fell behind and the calves showed increased incidence of 

pulmonary diseases. In September 2006, three calves were sent to the 

Clinic for Ruminants for examination. Later the farm was visited. The 

general condition of several animals of the age of 1 day to 6 months 


was poor, the animals were emaciated, their development was reduced 

and their respiratory rate severely increased. Blood samples were 

taken from animals of different age groups for examination of 

glutathione-peroxidase activity (GSH-Px), thyroxin (T4) and selenium 

(Se) concentrations. 3 animals admitted to the clinic showed poor 

general condition, emaciation and severe pathological lung findings. 

Two of the calves had visible and palpable masses at the ventral aspect 

of the larynx. A main finding in blood examination was very low 

GSH-Px (12 – 42 U/g Hb). These animals were euthanized and 

submitted to post-mortem examination. All three animals had masses 

in the area of the larynx in size of 7-15 cm in diameter. Additionally, 

the animals had severe bronchopneumonia, and one animal also had 

fibrinous pleuritis. Histological findings included hypertrophy and 

hyperplasia with hyperaemia of the thyroid gland. The bacteriological 

findings were Pasteurella multocida in the lungs of two of the animals 

and BRSV-antigen in one animal. On the farm, nine calves had 

palpable masses in the area of the larynx of up to 10 cm in size. T4 

levels were very low in three out of nine animals (< 0.5 µg/l) and the 

GPX was below 8 U/gHb in all animals examined. Analysis of the 

grass silage, fed ad libitum, yielded an iodine concentration of 0.29 

mg/kg. Mineral feed was offered to the animals; however, it did not 

contain any Se or iodine. Another one day old calf with goitre, Sedeficiency 

and bronchopneumonia was treated in the clinic with 

potassium iodide, selenium, vitamin E and antibiotics. After 4.5 

months the calf had recovered and was discharged. It was concluded 

that the farm had an iodine deficiency problem, with clinical signs of 

goitre, Se- deficiency and bronchopneumonia. There are no 

restrictions for mineral supplements in commandments for organic 

farms, especially for selenium or iodine. In regions with iodine 

and Se- deficiency in the soils there should be a special attention to 

mineral supplements. 

238 Dynamics of Bone Metabolism in High Pregnant Heifers and 

First Lactation Cows 

J. Staric, M. Nemec, J. Jezek, M. Klinkon, T. Zadnik 

UL, Veterinary faculty, Clinic for Ruminants, Ljubljana, Slovenia 

Bone metabolism can be assessed by using biochemical markers of 

bone metabolism. Biochemical markers of bone metabolism are 

degradation products, by products of formation and enzymes involved 

in bone degradation or formation. They indicate whether bone is 

forming, degrading or remodelling more intensively. 88 late pregnant 

HF heifers and first lactation cows in different physiological periods, 1 

month before calving, up to 10 days before calving, within 48 hours 

after calving, 10 to 20 days in milk and at the peak of lactation were 

involved in the study. The study was performed during winter, when all 

animals were housed. Venous blood samples for measurement of bone 

specific alkaline phosphatase (BALP), C-terminal telopeptide crosslinks 

of collagen I (CTx) and total calcium (tCa) were taken according 

to the protocol. Mean serum tCa has the lowest value in a group within 

48 hours after calving, 2,12 ± 0,09 mmol/L and highest during the peak 

lactation, 2,56 ± 0,13 mmol/L. Interestingly the mean BALP has the 

highest value in a group within 48 hours after calving, 37,8 ± 7,9 U/L 

and the lowest value during the peak lactation, 22,3 ± 4,53 U/L. The 

highest mean CTx value was measured in group of cows 10 to 20 days 

after calving, 1,152 ± 0,292 ng/L and the lowest in heifers up to 10 days 

before calving, 0,513 ± 0,197 ng/L. All mentioned differences were 

also statistically significant at p

2 

Freie Universität Berlin, Veterinary Clinical Sciences, Berlin, 

Germany 

3 

Miner Agricultural Research Institute, Chazy, United States 

4 

University of Illinois, animal sciences, Urbana - Champaign, United 

States 

Hypophosphatemia and disturbed liver function are common findings 

in dairy cows during early lactation. Low serum phosphorus (P) 

concentrations have been associated with altered liver function and 

decreased regenerative capacity of the liver. To our knowledge, 

neither the liver P content nor the relationship between serum P 

concentration ([P]) and liver P content have been studied in 

periparturient dairy cows. We therefore investigated the change 

in hepatocellular P content in dairy cows from late gestation to early 

lactation. Liver biopsy samples of 6 dairy cows were obtained at -65, 

-30, -14, +1, +14, +28 and +49 days relative to calving. Amounts of 

P, potassium (K) and magnesium (Mg) in liver were determined using 

inductively coupled plasma-mass spectrometry and expressed as 

mg/g or µg/g liver dry weight. The amount of DNA in the biopsy 

sample was determined in order to explore the potential confounding 

effect of changes in cell volume on liver P content. Serum [P] was 

measured at -50, -36, -14, +1, +14, +28 and +49 d relative to calving. 

The amount of P in wet weight liver (PW), non fat wet weight 

(PNFW) liver, and per cell (indexed to DNA; PDNA) was calculated; 

similar values were calculated for K and Mg. Repeated measures 

ANOVA was used to identify changes in measured and calculated 

values over time. Liver P decreased significantly from 12.4±1.1 mg/g 

at -14d to 8.2±1.4 mg/g at +14d, increasing thereafter to 11.7±1.5 

mg/g at +49 d. Similar changes in dry weight liver were observed for 

Mg and K. PW and PDNA showed the same pattern of change over 

time with similar trends for the Mg and K content in wet liver tissue 

as well as indexed to DNA. In contrast, the amounts of K and Mg in 

non fat wet tissue remained constant over time whereas a significant 

decline of PNFW at +14d was determined. Serum [P] tended to 

increase from calving to +14d and did not reflect changes in liver P 

content over time. Our findings indicate that time-dependent changes 

of the electrolyte contents in dry liver weight are largely due to 

changes in hepatic water and fat content. As indicated by the P, Mg 

and K content in non-fat wet liver weight, the cytosolic 

concentrations of Mg and K were stable throughout the study period 

whereas the cytosolic P concentration significantly declined at +14 d. 

Our findings further indicate that serum [P] cannot be used to 

estimate liver P content. 

282 Effect of Vitamin D on the Bone Metabolism of Dairy Cattle 

with Hypophosphatemia 

JT. Wang, JL. Li, SW. Xu, FQ. Shi 

Northeast Agricultural University, College of Veterinary Medicine, 

Harbin, China 

The morbidity of mineral metabolic obstacle in cows was very high in 

Daqing and Anda regions in China, especially in some areas of Anda in 

which most areas belonged to alkaline soil which was composed of 

high content of NaHCO 3. There was 5mg/kg available phosphorus in 

soil roughly. Dairy cattle in these areas were found to have 

hypophosphatemia with detecting the content of phosphorus in serum, 

and they showed dyskinesia and low productive ability for long time. 

The aim of this study was to find out the method which prevented and 

cured dairy cattle with hypophosphatemia, and illuminated the effect of 

vitamin D on the bone metabolism of cows with hypophosphstemia. 30 

cows with obvious clinical symptoms which were hypophosphatemia, 

dyskinesia was assigned randomly. 10 cows as control feed low phosphorus 

grass (p0.2%), 10 cows as group 2: feed high phosphorus grass (p>0.2%), 

and got vitamin D3 (2.5x10 6 IU, weekly) by intramuscular injection and 

one capsule of vitamin D 3 (10mg/500kg, daily) by oral. Blood samples 

(serum) were withdrawn on 1d, 30d, 60d, 90d and 120d. The content of 

vitamin D (including vitamin D, 25-OH-D, 1,25-(OH) 2-D), calciumphosphorus- 

CT and BGP in the serum of group 2 were higher, and 

APK activity was lower than control and group 1. However, these items 

are near between control and group 1: the content of serum protein, 

GOP and GPT activity in serum had no difference in three groups. 

After 120 days the clinical symptom of 10 cows in group 2 was 

obviously slight, 10 cows in group 1 were better than control. These 

results clearly demonstrated that the content of calcium-phosphorus- 

CT, BGP, vitamin D3 and its metabolism activity in serum of the cows 

with hypophosphstemia were changed obviously with adding vitamin 

D and high phosphorus grass. With enhanced the content of calcium, 

phosphorus, CT and BGP in serum, vitamin D could effect the bone 

metabolism of dairy cattle with hypophosphstemia, and prevented 

dairy cattle hypophosphatemia 

Key words: cows, hypophosphstemia, vitaman D, bone metabolism 

283 Changes T3 and T4 Plasma Concentrations in Dairy Cows 

during Lactation 

K. Sinka, J. Illek, D. Kumprechtova, P. Novak 

Veterinary and Pharmaceutical University Brno, Clinic of Ruminant 

Diseases, Brno, Czech Republic, Czech Republic 

Objective: The objective of this study was to determine plasma 

concentrations of T3 and T4 hormones in dairy cows in preparturient 

period (20 to 10 days prepartum), in early lactation (15 to 20 days in 

milk -DIM), peak lactation (60 to 80 DIM), and late lactation (220 to 

250 DIM). 

Material and methods: The study was performed in a Holstein herd 

with average milk yield of 10,650 kg per 305 day lactation. Cows 

were housed loose in pens and fed total mixed ration consisting of 

maize silage, clover-grass silage, meadow hey, straw and 

concentrate. Proportions of TMR components and nutrient contents 

reflected the lactation phases and were in accordance with standard 

recommendations. 

Selected cows from different phases of calving to calving interval were 

withdrawn blood from the coccygeal vein, using the HEMOS tubes 

with heparin. Group A: late pregnancy cows (10-20 days prepartum), 

Group B: early lactation (15- 20 days prepartum), Group C: peak 

lactation (60-80 DIM), Group D: late lactation (220-250 DIM). In each 

group 10 cows without clinical disease signs and in optimum body 

condition were monitored. Hormones T3 and T4 were measured by 

chemiluminiscence, using the automatic analyzer IMMULITE 1000. 

Results were statistically evaluated. 

Results: The late pregnancy cows (A) showed the lowest T3 

concentrations and the highest T4 concentrations. In early lactation there 

was a significant decrease in T4 and non-significant increase in T3. In 

peak lactation, T4 levels were still low but non-significantly higher than in 

the early lactation. In late lactation there was a significant increase in T4 

and significant decrease in T3. The late lactation values were similar to 

those measured in late pregnancy cows. 

Table 1. T3 and T4 plasma concentrations (nmol/l) 

Group A B C D 

T3 T4 T3 T4 T3 T4 T3 T4 

X 1.71 74.64 1.93 36.9 1.95 43.96 1.77 63.96 

S.D. 0,179 8,562 0,183 3,157 0,135 7,096 0,095 7,592 

C.V. 0,105 0,115 0,095 0,086 0,069 0,161 0,054 0,119 

T-test A-B A-B B-C B-C C-D C-D 

** *** N.S. ** *** *** 

(P

Materials and Methods: Serum samples from recumbent cattle were 

selected from samples submitted to a laboratory in Germany for 

analysis. A questionnaire was completed in order to obtain data on the 

duration of recumbency, time of sampling, breed, and treatment 

outcome. Serum concentrations of Ca, Phosphorus, Mg, Na, K, total 

bilirubin, urea, and total protein were measured, and serum activities of 

AST, CK, and GLDH were determined. Analysis of variance was used 

to compare serum values of cows that did not recover and were 

sampled before (group NR1) and after (group NR2) their first treatment 

for recumbency, and recovered cows that were sampled before (group 

R1) and after (group R2) their first treatment, respectively. Nonnormally 

distributed variables were log transformed before analysis 

and P

lipases (porcine pancreatic lipase type II and type VI-S, microbial lipase), 

detergents (SDS 10%; Lubrol), and incubation periods (0.5-16 h) were 

tested. Triolein (Sigma, Steinheim, Germany) was used as internal 

standard. The coefficient of variance (CV) of TL analysis in the ten liver 

bioptates (range: 40-314mg/g FW) was in average 2.2% (max. 4.5%; 

three repetitions). For the extra incubation step prior to the final enzymatic 

TAG analysis (MTI diagnostics, Idstein, Germany) microbial lipase, 

Lubrol, and 16 hours of incubation provided best results. Mean recovery 

of Triolein was 101% (97%-107%) with mean intra and inter day (10 

samples, 5 repetitions) CV% of 0.75% and 2.7%, resp. The liver TAG 

(range: 4-260 mg/g FW) analysis showed mean intra and inter day CV% 

of 2.5% (0.35-5.6%) and 3.4% (2.3-4.9%), resp. The presented combined 

method for TL and TAG determination in small amounts of bovine liver 

tissue was simple, accurate and reproducible. 

This work was supported by WILHELM SCHAUMANN STIFTUNG, 

Hamburg, Germany. 

288 Study on the Mechanism in the Damage of Erythrocyte 

Membrane in Low-phosphorus Cows 

SW. Xu, FQ. Shi, DW. Xuan 


Harbin, China 

Objectives: Investigate the mechanism in the damage of erythrocyte 

membrane (EM) in low-phosphorus cows. 

Materials and Methods: These cows with field cases were divided into 

three groups, including hemoglobinuria group (HG), low-phosphorus 

group (LPG) and control group (CG). The phospholipid composition, 

skeletin, antioxidant function and shape of EM were determined. 

Results: The phospholipid composition, skeletin, antioxidative function 

and shape of EM obviously changed in HG and LPG. (1) Phosphatidylethanolamine 

(PE) content in HG was significantly lower than 

that in LPG and CG, but sphingomyeline (SM) and phosphatidycholine 

(PC) + phosphatidylserine (PS) content in HG was significantly higher 

than that in LPG and CG. In comparison between LPG and CG, PC + PS 

content were lower and SM content was higher in LPG. Significant 

positive correlation and negative correlation were observed between 

serum phosphorus and PE content, serum phosphorus and SM content, 

respectively. (2) There were no difference was found in EM skeletin 

between LPG and CG. Spectrin I, spectrin II, and band IV-2 content was 

lower in HG than that in LPG and CG, but band III was higher in HG than 

that in LPG and CG. (3) SOD activity and GSH-Px activity in HG and 

LPG was significantly lower than that in CG. MDA content in HG and 

LPG was significantly higher than that in CG. There were a significant 

positive correlation between serum phosphorus and erythrocytic SOD 

activity and GSH-Px activity, and a negaitive correlation between serum 

phosphorus and erythrocytic MDA content. (4) Through the observation 

of scanning electron microscope, the course of erthrocytolysis attributable 

to low phosphorus intake was proved: the erythrocytes changed from 

biconcaval shape to acanthocytes, spherocytes and to be destroyed 

eventually with the decresed serum phosphorus content. Treatment with 

phosphorus preparation could signficantly alleviate the change in 

erythrocyte shape and make it to return to normal shape. 

Conclusions: The phospholipid composition, skeletin, and shape of EM 

changed, and the antioxidant function of EM decreased in lowphosphorus 

cows. These further caused the occurrence of the haemolysis. 

Key words: cow; low-phosphorus; erythrocyte membrane 

289 Prevalence of Digital Dermatitis in First Lactation Cows 

Presented at Auction 

M. Hulek 1,2 , I. Sommerfeld-Stur 3 , J. Kofler 1 

1 University of Veterinary Medicine Vienna, Department of Horses 

and Small Animals, Clinic of Orthopaedics in Large Animals, 

Vienna, Austria 

2 Veterinary Practice Michael Hulek,Oberneukirchen, Austria 

3 University of Veterinary Medicine Vienna, Department of Animal 

Breeding and Reproduction, Institute of Animal Breeding and 

Genetics, Vienna, Austria 

Objective: The risk of introducing digital dermatitis (= DD) to dairy 

herds by the introduction of infected, newly purchased cattle is 

assumed to be considerable. The aim of the study was to assess the 

prevalence of DD in first lactation cows (FLCs) presented as breeding 

cattle at the monthly auction in one auction centre in Austria over a 

period of 10 months. 

Material and Methods: The FLCs to be examined were selected 

randomly for claw examination for each auction from the monthly auction 

catalogue. At each auction a minimum of 25% of FLCs were selected, 

with an average of 36% of all FLCs of all 10 auctions. After obtaining 

owner’s consent, the hindclaws were examined in a walk-in crush for DD. 

DD lesions were evaluated by the parameters localisation (plantar, 

interdigital, dorsal aspect), diameter in cm and lesion type (M1-M4). 

Other claw lesions and trimming status of the claws were also noted. 

Results: From a total of 1110 FLCs registered for the ten auctions on 

the catalogues, 399 were chosen for examination, and of these 199 

FLCs could be examined. In 63 cows from the random sample the 

owners did not consent to the examination. A total of 24 FLCs were 

found to have DD lesions on one or both hindlimbs, with at least one 

cow detected at nine of the ten auctions. The size of lesions ranged 

from 0.5 to 3 cm in diameter. The prevalence of DD determined at 10 

auction dates within 10 months was 12.06%. 

Conclusions: The results of this study show that about 12% of FLCs 

presented as breeding cattle at auction were affected with DD. This 

result suggests that in the worst case, if each of these FLCs affected 

with DD is introduced into a DD free herd, 24 new herds could be 

infected by DD during a 10 months period from one single auction 

centre in Austria. An additional risk is the infection of other animals at 

the auction from the use of shared walkways and pens, with a 

potentially much larger number of herds affected at a later stage. In 

order to reduce both these risks, we recommend that dairy farmers 

purchase only cows free from digital dermatitis. An examination of the 

claws for the presence of DD lesions should be carried out in all cows 

one or two weeks before they are presented at auction. This 

examination should be performed by trained experts and the findings 

should be documented in a special protocol, which should be presented 

with the cow at the auction, and be available for inspection by all 

potential buyers. 

POSTER ABSTRACTS 

443 Influence of Age, Season and Physiological State on some 

Biochemical Parameters in South Eastren Algerian Desert 

Goats 

H. Nadia 1 , M. Toufik 1 , M. Bakir 2 , B. Mabrouk 1 

1 Batna University, Veterinary, Batna, Algeria 

2 Batna University, Virology, Batna, Algeria 

Blood plasma Ca, P, Mg, Na, K, and Fe were analysed in order to 

establish biochemical references, and to study the influence of age, 

season and physiological state on the variation of these parameters. 

The results demonstrate: 

– Ca, Mg, and Na levels were high at birth and decrease with age. 

– The season had a signficant effect on the levels of these ions, for 

example Ca, Mg, K decreased and inversely Na increased during dry 

season. 

– The values obtained for the plasma Ca, Mg, Na, K, Ca:P and Fe levels 

in pregnant goats were 80.02±4.84 mg/l, 22.14±1.61 mg/l, 142±1.73 

mEq/l, 6.43±0.40 mEq/l, 1.29±0.37 and 91.11±18.84 µg/100ml 

respectively. They were significantly higher than in non pregnant and 

lactating females. 

– Ca (77.67±3.20 mg/l), Mg (20.44±1.66 mg/l), K (5.65±0.57 mEq/l) 

and Ca:P (1.35±0.35) were lower in lactating goat compared to 

pregnant and non pregnant goats. 

These values of plasma minerals can be used as reference to detect 

metabolic and/or nutritional disorders in goat. 

Key words: mineral metabolism, goat, lactating, desert 

44 The Evaluation of Vitamin A and ß-carotene Levels during 

Postpartum Period in Semi Industrial Dairy Farms in Iran 

M. Tehrani Sharif 1 , R. Mozaffary 2 , J. Gholami Seyed kolaee 3 , 

M. Rezaee ghale 2 , A. Cheraghzadeh 2 

1 Islamic Azad University , Garmsar branch, Department of 

Pathobiology , Faculty of Veterinary Medicine, Garmsar, Iran 

2 Islamic Azad University , Garmsar branch, Student, Garmsar, Iran 

3 Shahid Chamran University, Student, Ahwaz, Iran 

Objective of study: Vitamin A, retinol, plays a vital role in vision sense. 

Due to the presence of large amounts of beta-carotene in cattle’s foods & 


vitamin A stores in liver, the hypovitaminosis is not provable. Increased 

secretion of vitamin A in milk during postpartum period, increases the risk 

of hypovitaminosis occurrence in cattle’s feed on a poor beta- carotene 

nutrition. The aim of this study was to evaluate the value of vitamin A & 

beta-carotene during postpartum period in some semi industrial farms 

with poor beta-carotene based nutrition in Ghaemshahr Iran. 

Material & methods: Blood samples were obtained from 88 dairy 

cows during postpartum period. The samples were centrifuged and 

froze pa at -20 °C. To prevent direct contact of samples with air and 

light, serum were preserved at dark pockets. Cows were classified at 9 

groups based on postpartum period (1,2,3,4,5,6,7,8,9 month after 

parturition). Vitamin A & beta-carotene were determined by 

spectrophotometric method. 

Results: The mean value of vitamin A & beta-carotene were 

38.06±9.16 µg/dl & 33.8±9.74 µg/dl respectively. The mean value of 

vitamin A in 1,2,3,4,5,6,7,8,9 month postpartum were 33.98±7.26, 

29.51±3.5, 35.23±1.44, 37.20±7.62, 38.74±9.06, 38.09±8.20, 

39.91±6.96, 42.89±4.87, 46.05±5.01 respectively. The mean value of 

beta-carotene in 1,2,3,4,5,6,7,8,9 month postpartum were 37.02±3.45, 

40.17±1.56, 26.79±3.40, 40.29±6.26, 18.07±4.02, 36.15±3.6, 

59.61±8.36, 36.31±7.31, 31.11±8.19 respectively. 

Conclusion: The mean values of vitamin A were normal in spite of 

consumption of poor beta- carotene based nutrition. It might be due to 

high level of liver stores. There were not significant differences in mean 

vitamin A during postpartum period. The mean beta-carotene were lower 

than reference ranges during post partum period. The decreased level of 

beta-carotene indicates the nutritional deficiency of beta-carotene. 

Key words: Vit A, beta carotene, postpartum period 

445 Comparison of Clinical and Laboratuar Responses for 

Laparotomic Omentopexy and Laparoscopic Abomasopexy in 

Cows with Left Abomasal Displacement 

K. Yigitarslan 1 , N. Yavru 2 , S. Avki 1 

1 University of Mehmet Akif Ersoy, Faculty of Vet.Med, 

DEPARTMENT OF SURGERY, Burdur, Turkey 

2 University of Selcuk, Faculty of Vet. Med., DEPARTMENT OF 

SURGERY, Konya, Turkey 

The objective of this study was to compare the metabolic responses and 

recovery period in cows with left abomasal displacement following by 

surgical treatment either with laparotomic omentopexy or laparoscopic 

abomasopexy. Forty cows brought to the Faculty of Veterinary Science 

(The Universities of Selcuk and Mehmet Akif Ersoy) and diagnosed with 

left abomasal displacement were used. The cows were divided for two 

groups. Twenty cows (group 1) were treated with laparotomic 

omentopexy and the other 20 were treated with laparoscopic 

abomasopexy. There were no statistically differences between both 

groups concern about sex, age, number of calving, time after parturition, 

diseases period, level of displacement, feces quantity, body temperature, 

heart frequency and serum lactate levels. Serum K and Cl levels were 

reached to normal levels in laparoscopic abomasopexy group at 72 nd 

hours postoperatively. The serum cortizol levels were maximum at 0 th 

hours both in laparoscopic abomasopexy and laparotomic omentopexy 

groups, and it was decreased under the preoperative levels after 24 hours. 

In cows treated with laparoscopic abomasopexy, the serum cortizol levels 

were statistically decreased at 24 th and 72 nd hours compared to the 

laparotomic omentopexy group (p

collected from each cow and tested for the presence of Betahydroxybutyrate 

(BHBA) - an indicator of abnormal fat metabolism using 

Ketolac ® and following manufacturer’s instruction. Appetite and bovine 

activity (ALPRO DELAVAL System) were evaluated for 10 days post 

enrolment (PE). Rectal temperature and symptoms of clinical ketosis, Left 

Displaced Abomasum or other diseases, were also recorded. Milk BHBA 

200 Ïmol/L was used as the cut-off value for diagnosis of SK while values 

andomly assigned to one of five treatments. Four and two weeks 

before expected calving, the heifers were injected intramuscularly 0 ml 

(T0), 10 ml (T1), 20 ml (T2), 30 ml (T3), 40 ml (T4) selenium and 

vitamin E respectively. The control (C) group received no supplement. 

Blood samples were collected from heifers four weeks before expected 

calving and at calving day. Blood samples of calves were drowning 

from the jugular vein at birth and 7 days of age. Serum concentrations 

of Se were determined using hydride generation Atomic absorption 

Spectrophotometer. The result indicated that the serum concentration 

of Se, Cu and Fe increased in T3 and T4 treatment heifers at calving 

day but no change seen in serum concentration of Zn. Serum Cu 

concentrations in calves of treated groups increased at birth and first 

week of age but was not significantly different. The serum 

concentration of Cu and Fe increased in calves of heifers given Se and 

vitamin E compound esp. in T3 and T4 treatments but the serum 

concentration of Zn decreased. It seems that the Zn: Cu and Zn: Fe 

ratios be disturbed which lead to Zn reduction. It can be concluded that 

Zn should be supplemented when more than 40 mg/Kg Dm 

administered to pregnant heifers. 

Key words: Zn, Cu, Se, interaction, heifers 

454 Effect of Mixed Enzymes on Alfalfa Hay Nutritive Value 

Using in vitro Gas Production and in situ Techniques 

B. Baghbanzadeh Nobari, A. Taghizadeh, H. Janmohamadi, 

S. Alijani, H. Paya, G. Moghadam 

Dept. of Animal Science, Faculty of Agriculture, University of Tabriz, 

Tabriz, Iran 

Fibrolytic enzymes are typically cellulase that degrade cellulose or 

xylanase that act on hemicellulose (Beauchemin et al., 1995). Enzymes 

applied to forages immediately before incubation enhance the digestion 

of dry matter (DM) and neutral detergent fiber (NDF) suggesting that 

fibrolytic enzymes applied at feeding can improve digestion in 

ruminants. 

455 Changes of Serum Parameters in Monensin-contained Diet of 

Gezel Sheep 

B. Baghbanzadeh Nobari 1 , A. Rezapour 2 , P. Namavar 1 , 

G. Moghadam 2 

1 

Dept. of Animal Science, Faculty of Agriculture, University of 

Tabriz, Tabriz, Iran 

2 

Dept. of Animal Science, Faculty of Agriculture, Islamic Azad 

University of Tabriz, Tabriz, Iran 

Five Gezel male lamb treated with 4 levels of monensin (0, 10, 30, and 

60 mg/kg of diet) in a change-over design. Blood samples were taken 

one hour before and 2 hrs after feeding. Serum was separated and 

freezed up to analysis. Serum levels of glucose, beta-hydroxy-butyrate, 

triacylglycerol, total protein, albumin (albumin/ globulin), and blood 

urea nitrogen (BUN) was measured by colorimetric methods. 

Considering liver health, serum activity of AST, ALP, and GGT was 

measured. Ruminal fluid sample was taken for pH analysis 

(ruminocynthesis). Total and differential white blood cell count and 

phagocytosis test undertaken. Results of this study showed that 

monensin in a level of at least 30 mg/kg of diet could effectively inhibit 

acidosis occurrence. Acidosis has a negative effect on appetite; acidotic 

animals have lower levels of glucose in serum. But, monensin does not 

have any independent positive effect on serum glucose. Even two-folds 

of this dosage had no negative effect on liver. Monensin does not any 

stimulatory effect on natural immunity either independently or dosedependently. 

Generally staying, monensin may be inhibiting acidosis 

elated complications such as natural immune depression. 

Key words: gezel, immunity, monensin, sheep 

456 D-vitamin Status of Cows in Preparturient and 

Postparturient Periods under Injection of Cholecalciferol 

L. Yuskiv 1 , B. Kurtiak 1 , V. Vlizlo 2 

1 Institute of Animal Biology UAAS, Cattle Nutrition, Lviv, Ukraine 

2 Institute of Animal Biology UAAS, Scientific Center for Prion 

Infections Study, Lviv, Ukraine 

The aim of our work was the investigation of influence of different doses 

(210 and 420 IU D 3/kg body weight) of vitamin D 3 on vitamins status of 

cows in preparturient and postparturient periods under the parenteral 


injection of vitamin D 3 once before calving and three times after calving 

with interval of seven days. The experiment was carried out on the three 

groups of cows of Black-and-White breed, 5-6-years-old age, in a winterspring 

period. The cows of first group, to which vitamin D did not 

injection governed as control. To the cows of 2-nd and 3-rd groups the 

vitamin D 3 was injected intramuscularly in abovementioned doses. In 

blood the level of 25-hydroxyvitamin D, the concentration of calcium, 

magnesium, inorganic phosphorus, the activity of total alkaline 

phosphatase and its isoenzymes at 3-5 days before calving, 5-7 days and 

55-60 days after calving were determined. It was established, that the 

content of 25-OH D 3 in the blood serum of cows of 1-st, 2-nd and 3-rd 

groups at 3-5 days before calving was correspondently: 18,75; 25,95 and 

32,85 nmol/l, and at 5-7 days after calving 15,83; 20,83 and 31,88 nmol/l. 

At 55-60 day after calving the concentration of 25-OH D 3 in blood serum 

of cows was the greatest and varied from 20,48 till 35,0 nmol/l. Thus in 

the cows blood of 2-nd and 3-rd groups before and after calving dose 

dependence increase of concentration of calcium of total, protein-linked 

and ultra-filtrates was observed. The concentration of magnesium in the 

blood of cows of 3-rd group in the 5-7 day after calving was statistically 

significantly higher about 19%, and in the 55-60 day about 6 %, 

comparatively with the control. At the same time, in the blood of cows of 

2-nd and 3-rd groups, comparatively with the control group, dose 

dependence increase of concentration of inorganic phosphorus and 

decrease of the activity of alkaline phosphatase and its isoenzymes (bone 

and intestinal) was observed. Parenteral injection of cholecalciferol to 

cows before and after calving maintained the level of 25-OH Vit D 3 and 

effected on the exchange of mineral components of blood, that prevents 

the decrease of their level in postparturient period. The character of these 

changes depend on the physiological state of cows and on the doses of 

vitamin D 3. 

Key words: cows, prepartum, postpartum, calcium, 25-hydroxycholecalciferol 

457 Clinical and Subclinical Cases of Phosphorus Deficiency and 

Hypophosphatemia in Egyptian Buffaloes 

H. Morsy 1 , A. Sheikh 2 , M. El-gamal 3 

1 

Faculty of Veterinary Medicine, Suez Canal University, Department 

of Animal Nutrition, Ismailia, Egypt 

2 

Faculty of Veterinary Medicine, Department of Animal Nutrition, 

Ismailia, Egypt 

3 

Faculty of Veterinary Medicine, Department of Animal Nutrition, 

Cairo, Egypt 

The correlationship between food intake, hypophosphatemia, blood 

contents, pregnancy and health condition were done in 35 bufalloes 2- 

4 months before parturation at 7-9 years old (9 healthy as a control,12 

with clinical and 14 with subclincal hypophosphatemia according to 

blood , serum and feed analysis and clinical examination). 

Analysis of feed stuff revealed a lower level of phosphorous, a higher 

level of calcium and molybdenum with a normal level of copper. 

Clinical cases of hypophosphatemia showed normal temperature, 

elevated pulse and respiration, sluggish rumen movement, depraved 

appetite, pica, dullness, emaciation, red haemoglobin urine and pale or 

sometimes icteric mucosus membrane. 

The subclinical cases showed a normal temperature, pulse, respiration, 

depraved appetite, dullness and sometimes locomotors disturbances. 

The blood level of hemoglobin content and PCV were significantly 

decreased in clinical cases only. 

Serum levels of inorganic phosphorous was highly significant 

decreased with a decrease of serum copper level in clinical cases and 

significantly decreased of phosphorous in subclinical cases. Serum 

calcium and molybdenum levels were significantly increased in clinical 

cases and slightly increased in subclinical cases. Serum magnesium, 

sodium, potassium and chloride showed a non significant changes in 

clinical and subclinical cases. 

Treatment with Sodium dibasic phosphate (60 gm diluted in 300 ml 

distilled water was given 4 times within 12 hours intervals, The first 

injection was intravenous, the others was subcutaneous) as well as 

daily 20 ml catozal intravenous and 250 gm bran and 10 gm copper 

sulphate per os in food for one week was satisfy for treatment the 

clinical cases. Subclinical cases were treated with oral daily of 30 gm 

sodium dibasic phosphate, 125 gm of bone meal and 2-4 kilogram of 

brain for a period of one month. Correction of feed supply was helpful 

and advised in both cases. It is concluded that diet, blood and serum 

analysis and the general health condition were helpful for diagnosis of 

clinical and subclinical hypophosphatemia in pregnant buffaloes,

correction of diet and phosphorus supplement was useful for treatment 

and prevention of hypophophatemia. 

458 Ketosis in Dairy Cows under Obligatory Reduction of Feed 

Intake 

A. Elsheikh 1 , T. Allam 2 , H. Morsy 3 

1 Faculty of Veterinary Medicine, Animal Medicine, Zagazig, Egypt, 

Egypt 

2 Animal Health Research Institute,Zagazig, Animal Medicine, 

Zagazig,Egypt, Egypt 

3 Faculty of Veterinary Medicine, Suez Canal University, Nutrition 

and Clinical Nutrition, Ismailia, Egypt 

Clinical findings, post-mortem, laboratory examination of blood serum 

and urine as well as milk production and body weight were done in 

lactating Friesian cows subjected to a reduction of food intake at the 

first and second month of parturition. 

Clinical symptoms included loss of body weight, dehydration, sunk 

eyes, reduction of milk production, crossing of fore limb, lameness, 

constipation, ketoneuria and acetone odour were appeared within 3-7 

weeks after restriction of food intake. 

Post-mortem of dead or emergency slaughtered cows showed a yellow 

gelatinous foamy fluid inside and around visceral organs, oedema of 

pericardium, and intestine. Enlargement greasy and fatty degeneration 

of liver which was easily perforated on pressure. 

Serum levels of ketone bodies and total lipids were significantly 

increases meanwhile serum levels of glucose, total protein, calcium, 

phosphorus and magnesium were significantly decreased. It is 

concluded that prolonged feed restriction causes chronic wasting 

form of ketosis, serous complication and severe economic loss 

among dairy cows due to sharp drop of milk production and loss of 

body weight. 

459 RBC and WBC Counts in Lactating Dairy Cows Affected by 

Subacute Ruminal Acidosis 

M. Morgante 1 , M. Gianesella 1 , C. Cannizzo 1 , P. Dalvit 2 , 

M. Gatto 1 , LM. Coppola 1 , L. Moro 2 

1 

Universita’ degli Studi di Padova, Dipartimento di Scienze Cliniche 

Veterinarie, Legnaro (PD), Italy 

2 

Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (PD), 

Italy 

The influence of subacute ruminal acidosis (SARA) on ruminal 

microbiology has received considerable attention, but less is known 

about systemic manifestations that arise from SARA probably because 

it is well known that variables such as breed, stage of growth, 

reproductive status, and stage of lactation have an influence on many 

blood parameters. The aim of this study was to investigate possible 

changes in some haematological parameters in dairy cows affected by 

SARA. During autumn 2007 the study was carried out on 12 highly 

productive farms, stationed in different zones throughout northern 

Italy. In all farms, 12 cows in early lactation, in good body condition 

and without clinical signs of disease were selected randomly from each 

herd, to perform rumenocentesis and obtain rumen fluid. Blood 

samples for hematological profiles were obtained from the same cows 

that had rumenocentesis performed by jugular venipuncture. The herds 

were divided into 3 groups depending on the mean rumen pH: group A 

farms with pH>5.8 (normal), group B pH between 5.6 and 5.8 (critical) 

and group C, pH

observed in group II of the animals treated with prebiotics. According 

to the data presenting in the literature ‚-glucans and tylosine are 

considered as powerful immunostimulators of macrophages and other 

immune effector cells (e.g. monocytes, neutrophiles, NK etc.) mainly 

by binding with their specific receptors (CR3, TLR, CD16/CD18 etc.) 

situated on a surface of the cells and activated them. 

Key words: tylosine, prebiotics, cytokines, calves 

462 Effects of Omentopexy on Tbars and Plasma Frap in Dairy 

Cows with Left/Right Abomasal Displacement 

P. Mudron 1 , J. Rehage 2 

1 

University of Veterinary Medicine, Clinic for Rminants, Kosice, 

Slovakia 

2 

University of Veterinary Medicine Hanover, Foundation, Clinic for 

ruminants, Hanover, Germany 

Objective of study: Higher levels of free radicals and reactive oxygen 

species can result in a damage of biological macromolecules and 

tissues. Therefore, the present investigation was aimed to study the 

effects of omtopexy stress on thiobarbituric acid reactive substances 

(TBARS) and ferric reducing ability of plasma (FRAP) in dairy cows 

suffering from the left and right abomasal displacement (LAD and 

RAD, respectively). 

Material and methods: Ten H-F dairy cows, (5.26 years), admitted for 

treatment of left abomasal displacement (omentopexy), were used in our 

study. Blood samples were drawn from the jugular vein prior to surgery, 

immediately and then 15, 30, 60, 90 minutes, and 2, 5, 10, and 24 hours 

after reposition of abomasum. In the study on the right abomasal 

displacement ten H-F dairy cows, (4.74 years), admitted for treatment of 

right abomasal displacement, were used. Abdominal surgery 

(omentopexy) was performed in a standing position 1-2 hours after 

admission. The average surgery duration was 26.1 minutes (without 

anaesthesia). Blood samples were drawn from the jugular vein in the same 

intervals as in the cows with the left abomasal displacement. 

Results: LAD Surgical stress resulted in a significant increase in plasma 

cortisol concentrations (p

extract (Mycosorb ® , Alltech Inc.) on milk quality (SCC, % fat & 

protein) and quantity. 

Materials and methods: The study was conducted on dairy cow herds 

where visual observations indicated the presence of mycotoxin 

contaminated forage. The study was conducted on 2 dairy farms. The total 

milk production and milk composition was recorded for the total trial 

period. The trial was conducted near Rome, Italy. The trial was done on a 

“before and after” basis with production figures being compared before 

and after the inclusion of Mycosorb Farm Pak. (including Mycosorb at 

20% on a carrier). Mycosorb Farm Pak was distributed at 100 g/head/days 

for the first 10 days and 50 g/head/day after until the end of the trial. 

Results: FARM 1The trial was conducted with 83 lactating Holstein- 

Friesian cows. The basal diet consisted of corn silage, Ryegrass hay, 

alfalfa hay, maize meal, feed concentrate (31% crude protein), soybean 

meal and peas. The total milk production and milk composition was 

recorded for 5 months: first 2 month without Mycosorb and 

consecutive 3 months with Mycosorb in the diet. Mycosorb inclusion 

increased milk yield by 1.5 l/h/d while stage of lactation remained 

identical; protein percentage (3.36% vs 3.41%) and fat percentage 

(3.71% vs 3.97%) while somatic cell count (SCC) was reduced (529 vs 

196, x1000 cells/ml). FARM 2The trial was conducted with 74 

lactating Holstein-Friesian cows. The basal diet consisted of corn 

silage, ryegrass hay, alfalfa hay, feed concentrate (21% crude protein). 

The total milk production and milk composition was recorded for 12 

months: 3 months without Mycosorb followed by 3 months with 

Mycosorb in the diet and then again 3 months without Mycosorb. With 

Mycosorb in the diet, SCC decreased (430 vs 288, x1000 cells/ml) and 

increased again (288 vs 598, x1000 cells/ml) after the removal of 

Mycosorb. Mycosorb inclusion increased milk yield by 2 l/h/d without 

change in stage of lactation at herd level. 

Conclusions: In conclusion, the inclusion of Mycosorb into the diet 

can be used to alleviate mycotoxins negative impact on productive 

performance and milk composition. 

Key words: dairy cows, milk production, mycotoxins 

466 Effects of Increasing Prepartum Dietary Protein Level Using 

Poultry by-product Meal on Productive Performance and 

Health of Multiparous Holstein Dairy Cows 

F. Kafilzadeh 1 , M. Yazdi 1 , H. Amanloo 2 

1 Razi University, Department of Animal Science, Kermanshah, Iran 

2 University of Zanjan, Department of Animal Science, Zanjan, Iran 

The effects of two levels of crude protein (CP) using poultry by-product 

meal fed during late gestation on the performance, blood metabolites, 

some reproductive parameters and colostrum composition of Holstein 

cows was studied. Sixteen multiparous cows 26±6 d before calving were 

assigned to one of the two diets in a randomized block design to evaluate 

the effects of two isocaloric diets containing either 14 or 16 % crude 

protein. Postpartum diet was similar in both groups through wk 3 of 

lactation. Yields of milk, protein, lactose, fat, and SNF were not affected 

by prepartum crude protein level. Colostrum composition (fat%, CP% 

and Total solids%) blood metabolites (Ca, Glucose, Total protein, 

Albumin, Globulin, Urea N and Cholesterol), some of the reproductive 

performance indexes (Pregnancy duration, Days open, First AI, service 

per conception, First service Conception rate%, and Pregnancy rate%), 

and disease incidence were not influenced by prepartum crude protein 

level. Colostrum fat percent was significantly affected (P

The aim of the monitoring was to compare the influence of fat 

supplementation in the form of full-fat soybean seeds and hydrolyzed 

palm oil as energy sources for the metabolism of dairy cows in early 

lactation. During the first 8 lactation weeks, 17 dairy cows, divided into 

experimental (E; n=8) and control (C; n=9) groups, were followed. The 

basic diet was the same for both groups. The differences were in the 

composition of the concentrate mixture, which had a balanced content 

of nutrients, but different source of fat. The group E received heattreated 

soybean seeds (22% in concentrates) and the group 

C hydrolyzed palm oil. The two diets started to be fed on the day of 

calving. Blood samples were taken weekly (1 st -5 th ) and 8 th weeks after 

delivery. At the end of the experiment was taken ruminal fluid. During 

the experiment BCS in both group worsened similarly. In group E 

decreased from 3.8±0.2 to 3.3±0.3 and in Group C from 3.5±0.4 to 

3.1±0.4. The BCS of the group E was on average by 0.3 point higher 

than in group C, but the difference was not significant. Despite this 

unfavourable factor, a better performance of hepatic metabolism was 

proved in the group E. In this group lower concentrations of nonesterified 

fatty acids was identified in 3 rd (0.96±0.38 vs. 1.76±0.63 

mmol.l -1 ; p

not influence the concentration of Zn in milk, but significantly influenced 

the Zn concentration in blood serum. The efficiency of different organic 

and inorganic forms of zinc was almost the same. 

The work was realized with the support of MSM Project No. 

6215712403. 

Key words: microelements, serum, Zn 

473 The Evaluation of Coagulation Profiles in Cows with 

Abomasal Displacement 

P. Sobiech, Z. Kuleta, J. Radwinska 

University of Warmia and Mazury, Department of Internal Medicine, 

Olsztyn, Poland 

Abomasal displacement occurs in high-yelding cows during early 

lactation. The most common form of this disease is left abomasal 

displacement, in which the abomasum is trapped between the rumen 

and the left abdominal wall. The purpose of the study was to evaluate 

coagulation profile in cows with this disorder. 

Material and methods: Tests were carried out on 20 cows of H-F breed 

(Holstein Friesian) divided into two groups: the first group 10 cows with 

left abomasal displacement, the second 10 healthy cows. Diagnosis of 

abomasal displacement was confirmed by abdominal ausculopercussion, 

rectal examination and verified by operation. Laboratory tests included 

marking, using CoA Chrom 3003 BIO-KSEL Apparatus, such 

parameters as: prothrombin time (PT), fibrinogen concentration (FIB), 

activated partial thromboplastin time (APTT), thrombin time (TT), level 

of antithrombin III (AT III) and level of D-Dimer. 

Results: Mean values of determined parameters amounted in cattle with 

abomasal displacement: PT 20,43 sec., FIB 7,99 g/l, APTT 47,31 sec., TT 

26,63 sec., AT III 81,33 %, D-Dim 849,33 µg/l. In healthy cows these 

parameters reached values as follows: PT- 25,12 sec, FIB 4,15 g/l, APTT 

33,71 sec., TT 24,84 sec., AT III 111,12 %, D-Dim 286,15 µg/l. 

Discussion: In this study it was found that cattle with left abomasal 

displacement had significantly higher fibrinogen level, APTT time and 

extremely high D-dimer level together with significantly lower AT III 

level. Obtained results indicated, that ill cows showed abnormal 

coagulation profile which reflect the occurrence of disseminated 

intravascular coagulation syndrome (DIC). The laboratory diagnosis of 

DIC is based on prolonged APTT, PT, hypofibrinogenaemia, decreased 

concentration of coagulation factors, increased D-Dim level and 

decreased AT III activity. A diagnosis of DIC is concerning when at 

least three tests in coagulation profile are abnormal. According with 

literature data, DIC is very common complication in cattle with 

abomasal displacement, especially with right displacement, which is 

usually accompanied by a degree of volvulus. 

Conclusion: During left abomasal displacement in cows occurred 

disseminated intravascular coagulation syndrome with different level 

of coagulation abnormalities. 

474 Bread Leftovers Utilization in Cattle Rations in Tadla Area 

A. Ilham, M. Mazouz, S. Alali 

Institut Agronomique et Vétérinaire Hassan II, Morocco, Pathologie 

Médicale et Chirurgicale des Ruminants, Rabat, Morocco 

The present study aimed to evaluate the importance of bread leftovers 

utilization by cattle producers in Tadla area in Morocco and to assess their 

nutritive value as well as their effects on dairy cow performances and 

diseases incidence. A follow up study was conducted during 3 months to 

compare the performances and the diseases occurrence in a sample 

consisting of 12 dairy farms among which 6 farms totalizing 51 cows used 

bread leftovers all the year around (F 1) and the 6 others composed of 53 

cows that have never used them (F 2). The main results obtained are as 

follows: 

- Among all cattle producers in Tadla area, 35.8 % supplemented the 

rations with bread leftovers from which 16.4 % used them all the year 

around and 19.4 % only during periods of feed shortage. 

- The total quantity of bread leftovers used per year in Tadla area was 

estimated to about 5230 metric tons. 

- The incorporation rate of bread leftovers in the concentrate portion of 

cattle rations varied from 20 to 100 % with an average rate of 30 %. The 

daily quantities used varied between 2 and 6 kg per cow with an 

average of 2.3 kg. 

- The composition per kg of bread leftovers was: 88.1% DM, 0.9% CF, 

11.9% CP and 3.6% Ash. The nutritive value was: 1.28 UFL (French 

dairy feed unit) / kg DM and 104.4g DCP / kg DM. 

- There was no significant difference between F 1 and F 2 regarding milk 

production and body live weight. However, the body condition score was 

in favour of cows of F 1 and the milk fat test was 1.27 % higher in F 2. 

- The incidence of health problems was higher in F 1 compared to F 2. 

Thus, abortion, acidosis and bloat rates were 15.7 vs. 7.4 %, 9.8 vs. 3.7 

% and 11.8% vs. 5% respectively in F 1 and F 2.65 

Key words: bread leftovers, nutritive value, dairy cattle, performances, 

health problems 

475 Influence of Diarrhoea on Chosen Indicators of the Blood 

Coagulation System at Dairy Cattle 

J. Radwinska, Z. Kuleta, P. Sobiech 



Objectives of study: In cattle breeding important is prevention and 

treatment of diarrhoeas. There are many factors causing diarrhoea. The 

aim research was evaluation of diarrhoea influence on chosen 

indicators of the blood clotting system at the dairy cattle. 

Material and methods: Research was passed on 2 groups of dairy 

cows. In group I were 16 cows with diarrhoea. Group II - 20 healthy 

cows (control group). The blood were taken from jugular vein. 

Coagulation factors were marked in blood plasma. To the laboratory 

analysis was used photometer Coag-Chorm 3003 (Bio-Ksel). During 

research were marked: prothrombin time(PT), thrombin time(TT), 

activated partial thromboplastin time(kaolin-kephalin time, APTT), 

fibrinogen, antithrombinIII(ATIII) and D-dimer. During research was 

not identify primary reason of diarrhoea. 

Results: There were changes in times length between ill and healthy 

cows. PT at cows with diarrhoea was longer than in healthy. There was 

an APTT extension (mean APTT at group I - 37,675s and group II - 

34,24s). In group I a mean TT was 31,39s in group II 26,44s. 

Fibrinogen concentration at ill cattle increased (6,61g/l) to healthy 

cattle (5,24g/l). Mean value of D-dimer in group I and II differed about 

103 µg/l. ATIII in each groups were: group I 108,7% and group II 

107,7%. PT describes exogenous tract and APTT characterize 

endogenous track of the blood clotting. The extension of PT is 

observed in deficiency of II, V, VII, X factors. All mentioned factors 

are dependent on vitamin K. A length of APTT is connected with II, V, 

X, VIII, IX, X, XI, XII factors. The PT prolongation might be 

connected with vitamin K malabsorption. This can be induced by 

changed bacterial flora in intestines (production vitamin K) or handicap 

absorption from alimentary canal (damages at intestine epithelium 

during infections). An increased D-dimer concentration (product of 

digestion stabilized fibrin) and TT prolongation (measures activity of 

fibrinogen) at the group I were connected with long PT and APTT. This 

can be an evidence of disseminated intravascular coagulation (DIC) in 

cow organism. Increased concentration of fibrinogen is DIC laboratory 

symptom. The activity of ATIII did not differ at sick and health cattle. 

Conclusions: Diseases with diarrhoea have influence on whole 

organism also on blood coagulation system. Abnormality in this system 

can lead to serious problems like DIC. 

476 Diagnostic Use of Pooled Metabolic Profiles in Czech Dairy 

Herds 

J. Davidek 1 , R. Van Saun 2 

1 Practitioner, Krasna Hora nad Vltavou, Czech Republic 

2 Penn State University, Veterinary & Biomedical Sciences, 

University Park, United States 

Study objective was to compare blood analyte concentrations collected 

at different periods relative to calving to determine diagnostic value in 

herd-based assessment of transition cow problems. In the course of 

clinical investigations on 37 dairy herds presented for various 

periparturient diseases or routine monitoring, blood was sampled from 

3 to 7 mature cows within defined time periods to run a pooled sample 

metabolic profile. Time periods were defined as (days relative to 

calving): Early Dry (>30 d prior), Close-up (

health group (HG) categories for analysis. Blood analytes were 

analyzed by ANOVA with main effects of period, HG and their 

interaction with herd as a covariate. Herds ranged in size from 100 to 

750 cows with Holstein and Simmental being the predominate breeds. 

A total of 40 metabolic profiles were performed. Herds were 

categorized into HG of no problems (NP, n=6), heat stress (HS, n=6), 

fresh cow diseases (FD, n=16), low production (LP, n=5) and other 

diseases (OD, n=7). Time period influenced (P

480 Mineral Metabolism during Late Pregnancy and Calcium 

Status after Parturition in Dairy Cows 

M. Casalone 1 , C. Cannizzo 1 , A. Stefani 2 , L. Moro 2 , 

M. Gianesella 1 , M. Morgante 1 

1 

Universita’degli Studi di Padova, Dipartimento di Scienze Cliniche 

Veterinarie, Legnaro (PD), Italy 

2 

Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (PD), 

Italy 

Milk fever represent one of the most important economic losses in dairy 

cows management, therefore to found some indicators (blood parameters 

or functional test) before calving should be very important for the 

prevention of hypocalcaemia and related disorders. The objective of this 

work was to study some blood and urinary excretion parameters during 

late pregnancy that could be related with the calcium status after 

parturition. 60 dairy cows from a farm located in the North of Italy with 

high incidence of milk fever were monitored 7 (±2) days before calving 

and 24h after calving: blood and urine samples were simultaneously 

collected for creatinine (Cr), calcium (Ca), phosphorus (P), magnesium 

(Mg), sodium (Na) and potassium (K) determination. Fractional excretion 

(%CrX) of electrolytes were calculated by the following formula: %CrX 

= Xu/Xp x Crp/Cru x 100 where “u” and “p” were the urinary and 

plasmatic concentration, respectively. Results were subject to ANOVA 

and correlation analysis using SIGMA STAT 3.05. Blood level of Ca after 

parturition show only a significant (P < 0.05) correlation (+ 0.490) with 

the blood level of Mg during late pregnancy but the results indicated that 

there were several significant correlation between the urinary level of Ca 

after parturition and some parameters during late pregnancy: in particular 

with the blood level of P (+ 0.378), with the urinary level of Ca (+ 0.384), 

P (+ 0.401) and Mg (+ 0.277) and with the fractional excretion of Ca (+ 

0.360) and P (+ 0.374). These data suggest that modifications of some 

blood and urinary parameters during late pregnancy could be related with 

the calcium metabolism after parturition. Although no animal developed 

any hypocalcaemia status post partum, this data suggest that it could be 

very important to do further studies especially on Ca and P excretion 

before calving to found useful indicators for risk evaluation of single cows 

regarding the probability to develop hypocalcaemia and related disorders 

after parturition. 

481 Effect of Glycerol-containing Preparation on Changes in 

Selected Parameters of Dairy Cows in Experimental and Field 

Conditions 

Z. Farkasova 1 , P. Reichel 1 , K. Kovacocyova 1 , M. Fabini 2 , 

A. Bobus 2 , G. Kovac 1 

1 University of Veterinary Medicine, Department of Internal 

Medicine, Kosice, Slovakia 

2 Polychem s r. o., polychem, Prievidza, Slovakia 

The aim of our study was to determine the effects of glycerol-containing 

preparation on changes in selected parameters in rumen fluid (volatile fatty 

acids - VFA, acidity - pH), blood serum (glucose, total cholesterol - TCH, 

total lipids - TL, triglycerides - TG, nonesterified fatty acid - NEFA, betahydroxybutyrate 

- BHB), and milk (fat, protein, urea) of dairy cows. The 

observation was aimed at prevention of lipomobilisation syndrome of 

cows during peri-partal period. In experimental conditions we determined 

optimal dose of glycerol from three selected concentrations. We used 150, 

200, 300 g/day/cow in 24 dairy cows. In the third experimental group 

(300g) the positive effects were recorded. The following observations were 

done in herd of 210 Holstein-Friesian dairy cows kept in loose housing 

with annual milk yield of 8500L. Glycerol bound to powder vehicle was 

added to the diet at the dose of 320g per animal and day for 35 days (3 

weeks before and 3 weeks after parturition). To compare glycerol effects 

on health and production, 12 experimental cows were selected and divided 

retrospectively by date of calving into two groups E1 (n = 6) and E2 (n = 

6), and by the period of glycerol intake before and after parturition (E1

lactating cows is sparse. Lotan and Adler (1976) reported that days 

open, number of inseminations, and pregnancy rates for 18 pairs of 

dairy cows having DP of 30 and 60 d were numerically similar. 

Remond et al. (1997) reported that BW of cows with no planned DP 

increased 24 kg in the first 60 DIM, whereas BW of cows with a 60d 

DP lost 28 kg during same time period. We speculated that cows 

with a DP of 56 d would be in more negative EB than cows with no 

planned DP due to reduced milk production in cows with no planned 

DP. Thus, cows with no planned DP were expected to ovulate earlier 

than cows with a 56-d DP; whereas cows with a 28-d DP were 

expected to be intermediate in days to first ovulation based on an 

expected intermediate level of negative EB. At the base of the 

importance of dry period management on the following milk 

production, mastitis rates, reproductive indexes and normal life of 

cows in after following parturition, the effects of different lengths of 

dry period on reproductive indexes were to be evaluated .One 

hundred Holstein dairy cows from 10/5/85 to 25/6/86 were selected 

from Bonyad Janbazan Shohada dairy farm in Razan Hamadan 

(Iran). In depended variable was the length of dry period and 

depended variables were reproductive indexes (1- parturition to first 

service, 2- days open, 3- numbers of service to pregnancy, 4mastitis 

rate on the first month of lactation). Control variables were 

1- pick of milk production, 2- BCS at the breeding, 3- number of 

lactation. Fisher Exact test and General Linear Model were used to 

statistical analysis. It is concluded that there was no correlation 

between dry period length and reproductive indices. 

Key words: dry period length, reproductive indices, cattle 

484 The Use of Calcium Chloride and Calcium Sulfate in an Oral 

Bolus as a Supplement for Post-partum Dairy Cows 

J. Spain 1 , J. Sampson 1 , L. Carstensen 2 , C. Jones 3 

1 

University of Missouri, Department of Animal Science, Columbia, 

United States 

2 

Boehringer Ingelheim Denmark, Division Vetmedica, Copenhagen, 

Denmark 

3 

Boehringer Ingelheim Vetmedica Inc, St. Joseph, United States 

Objective: The onset of lactation results in a sudden demand for 

calcium and imposes challenges to calcium homeostasis in cows. Cows 

unable to adapt to the calcium demand, develop hypocalcemia 

(incidences 23-39 %), which may manifest itself as milk fever (5-10%). 

This also increases the risk of periparturient disorders. With systemic 

acidification, PTH sensitivity is increased resulting in enhancing 

calcium mobilization from bone to blood. When the systemic pH is 

effectively decreased, urine pH values (pH 7.8) will decrease to a range 

between 6.2 and 6.8. The objective of this study was to investigate the 

effects of Bovikalc ® (Boehringer Ingelheim, St. Joseph, USA), an oral 

calcium supplement containing CaCl and CaSo4, on calcium 

homeostasis during the first 24 h after calving. 

Materials and Methods: Multiparous Holsteins (n = 20) were blocked 

by parity and day of calving and randomly assigned to either Control 

(C) group receiving no calcium supplement or treated (B) group 

receiving one bolus directly after calving and a second bolus 12 h later. 

Blood samples were drawn at 48 and 24 h before calving. Ionized blood 

calcium (iCa) was measured using an IDEXX Vet Stat Analyzer. Cows 

with an iCa level of 1.10 mmol/L or less were included in the study. 

Blood and urine samples were collected at 0, 1, 6, 12, 13, and 24 h post 

partum. Blood iCa and pH and urine pH were measured. 

Results: No differences in iCa levels were found between C and B 

groups (1.24 vs. 1.22; 1.19 vs. 1.18 mmol/L) at -48 and -24 h, 

respectively. iCa levels were similar at calving (0h, 0.95 vs. 0.94 

mmol/L) for C and B, respectively (p = 0.84). Thereafter, iCa levels 

were significantly higher in cows given the bolus as described (p = 

0.02). Urine pH differed with treatment (p = 0.002) and by treatment 

over time (p = 0.003). It decreased in B cows from 7.58 (0h) to 6.79 

(24h) compared to 8.00 (0h) to 8.09 (24h) for C cows. Blood pH did 

not change (p = 0.27) which reflects a tight homeostatic regulation of 

blood pH. Calcium supplementation with Bovikalc ® after calving 

decreased urine pH and increased iCa levels compared to control 

cows. 

Conclusions: By 2 administrations of Bovikalc ® with its two calcium 

components it was possible to increase the iCa level in blood of 

hypocalcemic cows significantly. Additionally it was shown that 

Bovikalc ® had a systemic acidifying effect which most likely enhances 

mobilization of the cow’s own calcium. 

Key words: dairy cows, hypocalcemia, calcium supplement 


485 The Microelements (MCEs) Supplementation by Mineral 

Lick (ML) and the Level of MCEs in the Blood, Colostrum 

and Milk of Dairy Cows 

S. Krys, A. Podhorsky, E. Lokajova, L. Pavlata 

University of Veterinary and Pharmaceutical Sciencies Brno, Clinic 

for Ruminants, Brno, Czech Republic 

The aim of this experiment was to verify the option of providing 

MCEs supplementation to dairy cows (DC) in the dry period through 

supplemented ML, and to then compare the content of MCEs in the 

blood, colostrum and milk of supplemented and of non-supplemented 

DC. The experiment was carried out on Czech Pied DC. We formed 

an experimental group (E, n=12), which was supplied with ML ad lib. 

enhanced with organic forms of MCEs (Se, Zn, Cu, Mn) for 4 to 5 

weeks before parturition and 1 week after parturition (a.p.). The 

control group (C, n=10) did not get any ML. Samples of blood were 

taken 14 days before calving, on the calving day, 4 th and 20 th day a.p. 

Samples of milk and colostrum were taken on the day of parturition 

(day 0) and then on days 1, 2, 3, 4 and 20 a.p. Reception of ML by DC 

was very uneven, its total consumption ranged from 0 to 0.25 

kg/animal/day. ML feeding did not result in evidential increase of 

concentrations of MCEs in DC blood. Only a trend to increased 

concentrations of Se and Cu in blood of E group was observed, where 

the average Se concentration increased by 60 µg/l (Cu by 9 µmol/l), 

while in C group, Se increased only by 30 µg/l (Cu by 6 µmol/l). 

Neither concentration of monitored MCEs in colostrum and milk 

pointed to a conclusive difference between groups (average 

concentration of MCEs in first colostrum in groups E and C was as 

follows: Se 115 vs. 102 µg/l; Cu 2.0 vs. 1.6 µmol/l; Zn 317 vs. 363 

µmol/l; Mn 0.6 vs. 0.4 µmol/l). The concentration of MCEs in 

colostrum a.p. dropped quickly. Significant drops were determined 

for Se, Cu and Zn on the first (for Mn on the second) day a.p. Apart 

from Mn, all monitored MCEs showed significantly lower (p200 µmol/l) 

and absence of clinical signs of ketosis are enrolled in the study and are 

randomized to one of three treatment groups: Two injections of 

Catosal ® (T1), one injection of Catosal ® and one of saline (T2), two 

injections of saline (T3), administered intravenously on Days 0 and 3, 

respectively, at a dose volume of 5 ml/100 kg bodyweight. Blood 

samples for assessments of changes in BHB, glucose, glutamate 

dehydrogenase (GLDH), and cholesterol are collected on Days 0, 3, 

and 6, respectively. From this study, a subgroup of 17 animals showing 

baseline serum BHB levels of >3.2 mmol/l and absence of clinical 

signs of ketosis was selected for an interim analysis. 

Results: The primary efficacy criterion was the treatment success, defined 

as the proportion of animals with serum BHB levels of >1.2 mmol/l on 

days 3 and 6 in each treatment group. Results showed that 30% and 60% of 

the animals in T1 were classified as treatment successes on days 3 and 6,

espectively. In contrast, none of the animals in T2 and T3 was classified as 

treatment successes. This effect was confirmed by statistically significant 

(p

491 Effect of Acarbose on Milk Yield and Composition in Early 

Lactation Dairy Cattle Fed a Ration to Induce Sub-acute 

Ruminal Acidosis (SARA) 

J. Keane 1 , L. McLaughlin 2 , P. Thompson 1 , G. Greenwood 2 , I. Bruce 1 

1 Pfizer Animal Health, Veterinary Medicine Research and 

Development, Sandwich, United Kingdom 

2 Pfizer Animal Health, Veterinary Medicine Research and 

Development, Kalamazoo, Michigan, United States 

SARA in dairy cattle is associated with decreased productivity, 

including decreased milk yield and milk fat. Metabolism of high 

carbohydrate rations increases the concentration of ruminal VFAs 

and, in severe cases, ruminal lactate resulting in ruminal pH below 

5.5 (SARA) or 4.5 (acute acidosis). Acarbose is a pseudotetrasaccharide 

that inhibits -amylase and glucosidase enzymes thus 

reducing the rate of carbohydrate digestion and has been 

demonstrated to maintain higher pH values by decreasing total VFAs 

and % propionate in an acute model of acidosis. To evaluate the 

efficacy of acarbose in managing low milk fat associated with SARA 

thirty early lactation Holstein cattle (average 101 days in milk) were 

fed a ration consisting of ground corn, soybean meal and corn silage 

(70:30 concentrate: fiber). The ration was shown to result in pH < 

5.5 for at least 4 hr/day for 80% of the days tested. Cattle were 

trained to feed from Calan gates and were assigned to control or 

0.75g/animal/day acarbose added to the total mixed ration (TMR) 

according to a cross-over design. Animals were fed the assigned test 

ration for 13 days and daily feed intake, milk yield and composition 

were measured for the subsequent 7 days of each 20-day cross-over 

period. Percent and yield of milk fat were increased in the acarbosetreated 

groups (3.33% vs. 3.04%, P

organs were identified. However, the results varied a lot within individual 

groups. Mean values of Cu concentration in individual groups ranged as 

follows (tissues are ordered in a descending line): liver (9.77-12.95 mg/kg 

of fresh tissue), heart (3.90-4.41 mg/kg), kidney (2.49-2.92 mg/kg), lungs 

(2.45-2.66 mg/kg), tongue (1.81-1.94 mg/kg), diaphragm (1.39-1.61 

mg/kg), pancreas (1.25-1.56 mg/kg), spleen (1.24-1.52 mg/kg), thigh 

musculature (0.85-1.03 mg/kg) and neck musculature (0.77-0.87 mg/kg). 

On the basis of our results Cu supplementation with concurrent 

administration of Zn, Mn and Se in higher doses does not cause different 

concentrations of Cu in individual tissues and organs of the goats. 

The work was realized with the support of MSM Project No. 

6215712403. 

Key words: microelements, liver, kidney, heart, lungs 

495 Evaluation of Copper Status in Dairy Cows Living in Region 

Irrigated with Potato Utility Refuses 

M. Kleczkowski, W. Kluci_ski, T. Jakubowski, K. Dembele 

Warsaw University of Life Sciences, Faculty of Veterinary Medicine, 

Clinical Sciences, Warsaw, Poland 

Copper status (CS) can be determined by 5 blood parameters: total 

plasma copper (Cu) zinc (Zn), molybdenum (Mo) as well as 

ceruloplasmin (Cp), and superoxide dismutase (SOD) activities. The 

aim of experiment was to evaluate CS from the region irrigated with 

potato utility refuses. The experiment lasted for 6 months, and it 

comprised 45 cows of the Black-White breed, at the 3 - 6 years. Cows 

were chosen from the 3 districts in the valley of Narew River, each 

group (1, 2 and 3) with a different irrigation method. Cows, from 1 and 

2- experimental groups, inhabited areas irrigated with utility refuses 

from potato industry company. Area inhabited with cows from 1 group 

was irrigated with utility refuses by using a flow method. Area 

inhabited with cows from 2 group was irrigated with utility refuses by 

using a sprinkle method. Cows from 3 group were control. Mineral 

analysis of utility refuses had been made before experiment started. 

Blood samples from all cows were collected 3 times: 1- one week 

before calving, two-three days after calving, and 3- six weeks after 

calving. Cow blood parameters that were determined are: Cu, Zn, Mo 

concentration, and Cp, SOD activity. Average value of particular 

parameters for cows from group 1 was: Cu- 8.99 µmol/ dm 3 , Mo -0.24 

µmol/ dm 3 , Zn- 11.46 µmol/ dm 3 , SOD- 70.11 U/gHb, Cp- 1.37 µmol/ 

dm 3 , for cows from group 2 was: Cu- 7.23 µmol/ dm 3 , Mo- 0.21 µmol/ 

dm 3 , Zn- 9.82 µmol/ dm 3 SOD- 54.22 U/gHb, Cp- 0.89 µmol/ dm 3 , for 

cows from control group 3 was: Cu- 12.07 µmol/ dm 3 , Mo- 0.26 µmol/ 

dm 3 , Zn- 14.55 µmol/ dm 3 , SOD- 81.36 U/gHb, Cp- 1.66 µmol/ dm 3 . 

The analysis of results showed that nutrition of dairy cows produce a 

decrease of copper status and Cu-dependent antioxidant activity in 

blood of cows. 

Key words: cows, copper, antioxidants, utility refuses 

496 Diurnal Variation of Acid Base Status in Cows Fed Different 

Anionic Salts Once a Day Only 

C. Gelfert 1 , A. Löptien 2 , N. Montag 2 , R. Staufenbiel 2 

1 

University of Veterianary Medicine, Clinic for ruminants, Vienna, 

Austria 

2 

Faculty of Veterinary Medicine, Freie Universitaet Berlin, Clinic for 

Ruminants and Pigs, Berlin, Germany 

Eleven non-lactating, non-pregnant, Holstein-Friesian-Crossbreed 

cows received 2,000 meq of either calcium chloride (CaCl 2) or calcium 

sulphate (CaSO 4) once a day for a period of one week. At day seven, 

blood and urine samples were taken every four hours starting at 6 

o’clock before feeding and ending the same time the next day. Before 

feeding, urine analysis of the CaCl 2-cows showed a compensated 

acidosis due to an reduced pH and low levels of net acid-base excretion 

(NABE). Feeding AS changed acid-base status (ABS) noticeably in 

both groups over time. All cows developed a strong metabolic acidosis. 

The strongest acid load was visible 18 hours after the morning feeding. 

Changes were greater in CaCl 2-cows and NABE differed significantly 

(p=0.037) between both anionic salt groups. Nadir of NABE was - 

41mmol7l in CaCl 2-cows and +4mmol/l in the CaSO 4-cows. The 

fractional clearance of calcium was higher (p=0.027) in the CaCl 2cows 

due to a higher calcium excretion via the kidneys. Serum calcium 

concentration did not show significant differences between both groups 

(p>0.05) but changed significantly during the day (p

499 Secondary Hepatogenous Photosensitization in Buffalos due 

to the Excessive Accumulation of Hepatic Copper 

A. Minervino 1 , R. Barreto júnior 2 , F. Rodrigues 1 , L. Reis 1 , 

R. Ferreira 1 , S. Headley 3 , E. Ortolani 1 

1 

College of Veterinary Medicine, University of Sao Paulo, 

Department of Clinical Sciences, Sao Paulo, Brazil 

2 

Rural Federal University of Semi-Arid, Mossoró, Brazil 

3 

Faculty of Veterinary Medicine, University of Helsinki, Helsinki, 

Finland 

Four 1.5 yr-old, male, murrah buffaloes, maintained during six months 

with high Cu intake and without direct solar exposure, were transported 

to regular farming conditions. Shortly after sunlight exposure, all 

buffalos demonstrated clinical manifestations characteristic of 

photosensitization: anorexia, apathy and severe cutaneous lesions. 

Blood samples were obtained before solar exposure (T0), during the 

clinical manifestations (T1), and after recovery (T2) to evaluate 

haematological (PCV, Hb, RBC and WBC) and biochemical (total 

protein, albumin, g-glutamil transferase (gGT), aspartate amino 

transferase (AST), creatine kinase (CK), urea, and creatinine) 

parameters. Skin samples obtained by incision biopsy at T0 and T1 

were processed for routine histopathology. Treatment consisted of 

removal from sunlight exposure and aqueous Zinc oxide solution. 

Three animals recovered fullly and 1 died. The hepatic Cu 

concentrations and gGT and AST activities before solar exposure were 

above normal values. For haematological parameters, only RBC 

demonstrated significant difference, being reduced at T1 and returning 

to normality at T2. A similar pattern was observed with the total protein 

and albumin; but an opposite trend occurred with CK activity. The 

buffalo that succumbed had the highest liver of Cu concentration and 

more extensive macroscopic skin lesions. At T0 the buffalos had 

generalized alopecia, but insignificant histological alterations 

(acantholysis and neovascularization). However, at T1 

histopathological examination revealed hepatic photosensitization 

characterized by diffused, moderate parakeratotic hyperkeratosis, 

multifocal moderate acantholysis, degeneration and necrosis of 

squamous epithelial cells, sweat gland atrophy, sebaceous gland 

hyperplasia, and dermal edema. Although the buffalos demonstrated a 

liver insufficiency at T0, the skin lesions indicative of secondary 

hepatogenous photosensitization were only triggered after the solar 

exposure. In this case, a high Cu intake induced hepatic lesions that 

may have caused inefficient metabolization of phylloerythrin resulting 

in secondary photosensitization after the sunlight exposure. 

Research supported by FAPESP 

Key words: buffaloes, hyperkeratosis, photosensitization, hepatic 

lesions 

500 Trace Elements in Cattle: Copper and it`s “Friends” 

A. Mueller, B. Freude, M. Scholz, M. Weiss 

IDEXX, Vet Med Labor GmbH, Ludwigsburg, Germany 

Prophylactic procedures to prevent diseases become more and more 

important in cattle rearing. Adequate supply in trace elements is 

required. Copper (Cu) deficiency is regarded as the second most 

common mineral deficiency of cattle in the World (Telfer et al, 1996; 

Black und French, 2004). Two different copper deficiencies are known: 

a primary copper deficiency induced by a simple deficiency of copper 

in the diet, and a secondary, resulting from the reduction in copper 

absorption or utilization by the antagonistic effects of molybdenum 

(Mo) and iron (Fe). 340 cattle samples were analysed as serum/plasma 

pairs regarding the concentration of Cu, Mo and Fe with ICP-AES 

(Inductively Coupled Plasma Atomic Emission Spectroscopy) during 

July 2006 until March 2007. The Cu-plasma analyses showed that only 

24% of these samples were below the reference value for Cu (80 

Ïg/dL). Compared to these results, 84% of the corresponding serum 

samples showed a Cu deficiency (below 80 µg/dL). Serum clotting 

reduces Cu by up to 80% compared to the plasma values (in 

confirmation to Laven & Livesey, 2006). In contrast to copper the 

analyses of Mo in bovine plasma and serum resulted in comparable 

values. 4500 bovine serum and plasma samples were analysed for Mo. 

80% of Mo-concentrations were below 26 µg/L. Only a few samples (< 

1%) showed higher values (max. 3300 µg/L). A correlation between 

low Cu and high Mo values in plasma, or low Cu values with high Fe 

concentrations could not be found. Additionally, hair analyses of Cu (N 

= 173) and Mo (N = 99) were performed. 85 % of the hair samples (3- 


times analyses of each hair sample) showed Cu values between 5 and 

20 mg/kg (49%: 6,6-10,4 mg/kg; 4 %: below 6,6 mg/kg), 15 % of the 

samples showed higher values up to 100 mg/kg). The Mo 

concentrations in the hair samples varied between 0 up to 800 µg/kg or 

even higher). The comparison between Cu and Mo in hair (N = 99), or 

Cu hair/plasma (N = 107) and Mo hair/plasma (N = 70) showed no 

correlation. The analysed cattle samples with low copper plasma 

concentrations showed mainly a primary copper deficiency. A 

secondary copper deficiency caused by a molybdenum intoxication 

could not be found in this set of samples. 

501 Effect of chronic glucose infusion on lactation performance 

and metabolic profiles in dairy cows 

B. Al-Trad 1 , K. Reisberg 1 , T. Wittek 2 , A. Alkaassem 2 , G. Gäbel 1 , 

M. Fürll 2 , J.R. Aschenbach 1 

1 Faculty of veterinary medicine, University of Leipzig, Institute of 

Animal Physiology, Leipzig, Germany 

2 Faculty of veterinary medicine, University of Leipzig, Medizinische 

Tierklinik, Leipzig, Germany 

Objective: To characterize the metabolic adaptations that occur during 

gradual increases of glucose supply via prolonged i.v. glucose 

infusions in dairy cows. 

Material & methods: Twelve midlactation dairy cows were assigned 

randomly to continuous i.v. infusions of either saline (control group, 

n=6) or 40% glucose solutions (treatment group, n=6) for 28 d. The 

infusion dose started at 1.25% of the daily energy (NEL) requirement 

and then gradually increased until a maximum dose of 30% NEL 

requirement was achieved at d 23. Then infusion dose was maintained 

at 30% for 5 d. The treatment (feed plus infusion) was hypercaloric but 

isonitrogenous. Liver and skeletal muscle biopsies were taken on d 0, 8, 

16, 24, and 32. Blood samples were taken every 2 d at 10:00 a.m. and 

additional 24-h blood samples (6-h intervals) were taken before each 

biopsy. Milk and urine samples were also taken on the biopsies days. 

Results: No changes occurred in daily feed intake and milk production, 

milk lactose and urea concentrations. Milk protein percentage and yield 

tended to increase during the high infusion dose (i.e. 20 & 30% NEL 

requirement). Decrease in milk fat percentage and yields were 

observed in both groups; however, the decrease was numerically higher 

in the treatment cows. Body weight and BFT increased in the treated 

group. Small and occasional increases in blood glucose and insulin 

concentrations were observed in the blood samples taken at 10:00 a.m. 

every 2 d. However, at infusion of 30% NEL requirement, five out of 

six treated cows had blood glucose concentrations >8 mM at 16:00 

p.m., i.e. 1 h postprandially. Glucose infusion decreased serum NEFA, 

BHBA and BUN concentrations. Serum liver enzymes, cholesterol, 

bilirubin, K and Ca concentrations were not affected by infusion. Less 

than 2% changes were observed in serum P, Cl and Na concentrations. 

Glucoseuria was detected during the maximum infusion dose. Liver 

glycogen increased gradually to reach plateau on d 16 before it fell 

back to baseline values on d 32. Skeletal muscle glycogen increased 

and liver total lipid tended to increase during the high dosage of 

glucose infusion. 

Conclusions: Dairy cows on an energy-balanced diet do not direct 

excess glucose to increased lactation performance. Excess glucose is 

transiently stored as glycogen in the liver and, predominantly, 

transferred to body fat. At high dosages, glucose is also stored in 

skeletal muscle as glycogen and excreted via the urine. 

502 Biochemical Profile of Cattle with Induced Hypocalcaemia 

and Subsequently Treated with an Enriched Calcium 

Solution 

R. Barreto júnior 1 , A. Minervino 2 , F. Rodrigues 2 , E. Meira júnior 2 , 

R. Ferreira 2 , A. Lima 3 , C. Mori 2 , E. Ortolani 2 

1 Rural Federal University of Semi-Arid, Mossoró, Brazil 

2 College of Veterinary Medicine, University of Sao Paulo, 

Department of Clinical Science, Sao Paulo, Brazil 

3 Vallée S.A., Sao Paulo, Brazil 

Twelve two years-old Holstein heifers were used to study the 

biochemical profile during the induction and recovery of 

hypocalcaemia. The picture was induced by continuous infusion of a 

EDTA solution (5% and pH 7.4) into the jugular vein at the speed of 

220 mL/h until the animals presented definitive clinical signs of 

hypocalcaemia such as sternal or lateral recumbency; then the infusion

was immediately stopped and the heifers were treated, during 30 min, 

with 1 mL/kg BW of a calcium solution (ValléeCálcio ® ) with 2.44 g of 

calcium /100 mL, from three sources: Gluconate, Lactate and Sacarate, 

enriched with 5 g Glucose, 0.47 g phosphorus and 0.19 g magnesium 

per 100 mL. Venous blood samples were taken before infusion (T0), 

when the muscle tremors started (T1), at the end of the infusion (T2), at 

the end of the treatment (T3) and 24 h later (T4) for serum analysis of 

calcium, ionized calcium, phosphorus and magnesium, plasma glucose 

levels and blood activity (30 °C) of g-glutamyl transferase (gGT), 

aspartate aminotransferase (AST) and creatine kinase (CK). The 

induction generated a similar clinical picture as seen in the natural 

course of hypocalcemia and the treatment with the calcium solution 

promoted total recovery. The induction generated a significant decrease 

in calcium, ionized calcium, phosphorus and magnesium levels, 

principally at T2, but the treatment transiently increased the calcium 

and ionized calcium at T3 and all of the variables returned to normality 

24 h later. The plasma glucose levels increased only after the treatment, 

but returned to normal at T4. There was no change in the gGT activity 

throughout the trial, but a significant increase of AST and CK at T4, 

indicating that some muscle damage occurred during the process. 

Key words: hypocalcaemia, calcium, phosphorus, treatment 

503 Clinical Picture of Induced Hypocalcaemia in Holstein 

Heifers Subsequently Treated with an Enriched Calcium 

Solution 

R. Barreto júnior 1 , A. Minervino 2 , F. Rodrigues 2 , E. Meira júnior 2 , 

R. Ferreira 2 , A. Lima 3 , C. Mori 2 , E. Ortolani 2 

1 Rural Federal University of Semi-Arid, Mossoró, Brazil 


Department of Clinical Sciences, Sao Paulo, Brazil 


Twelve two years-old Holstein heifers were used to study the clinical 

picture during the induction and recovery of hypocalcaemia. The 

picture was induced by continuous infusion of a EDTA solution (5% 

and pH 7.4) into the jugular vein at the speed of 220 mL/h until the 

animals presented definitive clinical signs of hypocalcaemia, such as 

sternal or lateral recumbency; then the infusion was immediately 

stopped and the heifers were treated, during 30 min, with 1 mL/kg BW 

of a calcium solution (ValléeCálcio ® ) with 2.44 g Ca (gluconate, 

lactate, and saharate), 5 g Glucose, 0.47g phosphorus and 0.19 g 

magnesium per 100 mL. A clinical examination was performed before 

infusion (T0), when the muscle tremors started (T1), at the end of the 

infusion (T2), at the end of the treatment (T3) and 24 h later (T4). The 

induction generated a similar clinical picture as seen in the natural 

course of hypocalcemia, but a constant and involuntary kinetic tongue 

movement cleaning the nostril and nose and causing a lip sound. Limb 

stiffness and uneasiness preceded the muscle tremor, and from this 

moment on the animals refused food. Then apathy, staggering and 

mydriasis became evident. These symptoms were followed by a rapid 

sternal and in most cases lateral recumbency. Even in this last position 

the heifers kept the head turned to the flank. At this moment the skin 

temperature at the lower limb and anal reflex were decreased; anuria 

and somnolence were evident. The heart rate and hypophonesis 

increased as higher was the EDTA infusion. Conversely, rumen stasis 

occurred in most cases at T1. Most animals had gas bloat. The 

treatment with calcium rapidly reversed the clinical picture, restored 

the heart rate and the rumen tonus and did not cause any adverse 

reaction within 48 hours after treatment. 

Key words: hypocalvaemia, symptoms, treatment, recovery 

504 Evaluation of Faecal Quality as a Diagnostic Tool in SARA 

Diagnosis in Dairy Cattle 

J. Tajik1, Mg. Nadalian1, A. Raoofi1, Gr. Mohamadi2, A. Bahonar1 

1 University of Tehran, Clinical Sciences, Internal Medicine, Tehran, 

Iran 

2 University of Tehran, Clinical Sciences, Tehran, Iran 

Objectives: Sub acute ruminal acidosis (SARA) is characterized by 

daily episodes of low ruminal pH between 5.5 and 5.0. This digestive 

disorder is the consequence of feeding high grain diets to dairy cows, 

which are adapted to digest predominantly forage diets. SARA may be 

a common and economically important problem in dairy herds. There 

is Limited knowledge of the symptomatology of SARA. Changes in 

faecal consistency and structure of SARA affected cows have been 

Scores 1 2 3 4 5 

Parameter 

Faecal Very Dry, stiff, Circular, Flat, Liquid 

consistency dry, semi moist raised loose, pools of 

(FCS) lumpy formed pat, symmetrical thinly faeces 

pats rings spread 

surrounding 

a dipped centre 

Faecal No few particles Massive 

undigested particles particles clearly presence 

feed particles visible of 

(FFP) particles 

Faecal gas No few Massive 

bubbles bubbles bubbles presence 

(FBS) of bubbles 

described. Considering the fact that SARA has to be understood as a 

transient condition with no clinical consequences visible at the time of 

insult, it seems that faecal changes rather apply to more severe states of 

ruminal acidosis than to SARA. This study was designed to evaluate 

the validation of faecal quality as a diagnostic tool in SARA diagnosis 

in dairy cattle. 

Materials and Methods: Ruminal fluid by means of rumenocentesis 

(Nordlund& Oetzel 1995) was collected from 117 cows (37 early 

lactation and 80 mid lactation cows) from 10 dairies, 2-4 hours 

following morning total mixed ration feeding. Additionally scoring of 

faecal quality (consistency, presence of undigested feed particles and 

gas bubbles) was done (table1). 

chi square and fisher exact tests were used in statistical analysis of data 

results. 

Results: There were no significant differences between individuals 

experiencing SARA (ruminal pH< 5.5) and the rest of population 

(ruminal pH>5.8) in FCS (P=0.908), FBS (P=0.577) and FFP 

(P=0.288). Also, no significant difference was found between animals 

with SARA and a marginal pH (5.6

they all died in a two to three day course. Of the necropsy findings, the 

severe fatty change of the liver seemed to be the most important. The 

diagnosis of “Primary pregnancy toxemia of Sheep” was made upon the 

gathered data. With this, the remaining ewes in the “non-pregnant 

diagnosed” group were rechecked for the presence of pregnancy and 

those found pregnant, were moved to a higher energy diet with 

nutritional supports and eventually further cases and losses were 

prevented. From these, it has been concluded that a small error in early 

pregnancy diagnosis via Ultrasonography can lead to significant losses 

in sheep flocks. So it is important to know, along with modernizing the 

techniques in animal husbandry, it is of great importance to implement 

the methods meticulously to prevent such problems. 

Key words: sheep, pregnancy toxemia, metabolic disorders 

506 The Use of Urinary pH Measured in Pooled Urine Samples 

for Monitoring the Use of Anionic Salts in Dairy Cow Herds 

C. Gelfert 1 , S. Römer 2 , N. Brand 2 , M. Goebbels 2 , R. Staufenbiel 2 

1 

University of Veterianary Medicine, Clinic for ruminants, Vienna, 

Austria 

2 

Faculty of Veterinary Medicine, Freie Universitaet Berlin, Clinic for 

Ruminants and Pigs, Berlin, Germany 

The use of pooled urine samples of dairy cows was evaluated in the 

monitoring of the use of anionic salts (AS). In a first step, it was evaluated, 

whether the pH of pooled samples reflect sufficiently the calculated mean 

of the pH measured in the single samples, which form the pooled one. 

Five large dairy farms in Germany were visited monthl, and 10 cows each 

from four different stages of lactation were randomly selected. From each 

cow, a urine sample was taken by using a urinal catheter and a pooled 

sample was produced for each stage of lactation by merging equal 

amounts of each single sample. The urinal pH was measured in the single 

samples and the pooled samples (x p) with a portal pH meter. After 

measuring the pH in the single samples, the mean pH of each lactational 

group (x s) was calculated. The evaluation of xp followed the procedure of 

Bland and Altmann (1986). The mean of xs was 8.22 and the mean of x p 

was 8.29. The urinary pH of Xp was slightly higher than xs. The mean 

difference was 0.07. These small increases of pH only bore a negligible 

risk of false diagnosis of urinary acid-base status. In the second step, the 

urinary pH of pooled samples was tested in reflecting the use of AS in 

dairy cow herds. 26 dairy farms were visited on which AS were used. 

Urine samples from 10 randomly selected healthy transition cows were 

taken, and a feed sample from the total mix ration including the anionic 

salts was taken to measure the concentrations of the following minerals: 

sodium, potassium, calcium, chloride, and sulfate, and DCAD was 

calculated. Although all farms used AS, a high variation of urinary pH 

was visible, ranging from 5.90 to 8.64. DCAD also showed a high 

variation ranging from -393 mEq/kg DM to 303 mEq/kg DM. A DCAD 

below zero was no guarantee for a decrease of urinary pH. Therefore, the 

farms were grouped according the diagnostic result of urinary pH (cut off 

point 7.8). DCAD tends (p=0.070) to result in lower values when urinary 

pH decreased (-1.96mEq&kg DM/114mEq/kg DM). Pooling samples of 

cows of the same stage of lactation minimizes laboratory work and 

resulting costs and the measured value corresponds sufficiently to the 

calculated mean of the single samples. The use of AS in the prevention of 

periparturient hypocalcaemia has to be monitored by urine analysis. 

Measuring DCAD may lead to misinterpretation when other faults in feed 

management exist. 

507 Effect of Anionic Salts Supplement in Last Two Months 

Pregnant on Metabolic Disorders and Blood Minerals of 

Holstein Cow 

M. Ganjkhanlou, A. Nikkah, A. Zali, M. Motlagh 

Animal Sciences Group, animal science, Karaj, Iran 

In order to, investigate the effect of feeding cow with different of dietary 

cation anion balance (DCAB) in far-off and close-up period, on 

metabolic disorders and blood composition of Holstein cow in 

pregnancy and fresh period this experiment was carried out. In this study 

18 Holstein pregnant cows (220-225 d) were fed individually, a base 

diet with tree different DCAB (+13(control), 0,-13 mEq [(Na+K)- 

(Cl+S)]/100gr dry matter) for 60±5 days. For decrease DCAB two anion 

salts such as ammonium chloride and ammonium sulphate were used. 

The result indicated milk fever percentage were 0, 33, 33 for diet -13, 

0,+13 mEq/100gr DM, respectively. The rate of retained placenta was 

9.37, 4.62, 4.61 in cows received diet +13,0,-13 mEq/100gr DM 


respectively. Blood pH, Na, K, Mg and Ca were unaffected by dietary 

cation-anion balance. Urinary PH of cows was affected by diets. Urinary 

pH increased linearly with increasing DCAB. The mean urinary PH was 

7.99 (for diet +13), 6.81 (for diet 0) and 6.11 (for diet -13). 

Key words: DCAB, ammonium chloride, ammonium sulphate, 

metabolic disorders 

508 The Effect of Sodium Selenite and Sel-Plex on Selenium 

Deficiency Prevention in Beef Cattle 

J. Illek 1 , M. Matejicek 2 , D. Kumprechtova 1 , M. Hulik 3 , T. Berka 3 



2 VVS, Vermerovice, Czech Republic 

3 Alltech, Alltech CZ, Praha, Czech Republic 

Objective: The aim was to investigate effects of dietary inclusion of 

different selenium (Se) forms and AA cows in the grazing season. 

Material and methods: 3 Se deficient herds, grazed at Se deficient 

pastures (bellow 0.1 mg Se/mg DM). Herd A got a complete mineral 

lick with sodium selenite (30 mg Se/kg). Herd B got mineral feed with 

sodium selenite (30 mg Se/kg). Herd C got mineral feed with organic 

Se (Sel-Plex) (30 mg Se/kg). Grazing season was April - November. 

There were groups by 40 cows, along with calves and a bull. They fed 

ad libitum on pasture forage and the mineral lick or mineral feeds only. 

In the end of the pasture season, 10 cows from each herd were 

withdrawn blood samples. Whole blood Se levels and 

glutathionperoxidase (GPx) activity were measured, metabolic profile 

parameters analysed in serum. In the end of grazing season, all the 

cows were pregnancy tested. 

Results: Se (ug/l) and GPx (ukat/l) in blood 

Herd A B C 

Se GPx Se GPx Se GPx 

Mean 52.0 aA 320.8 A 61.4 bA 494.6 B 84.4 B 796.0 C 

S.D. 4.5 97.8 3.9 44.2 11.8 71.0 

C.V. 8.7 30.5 6.3 8.9 14.0 8.0 

a,b,c P < 0.05; A,B,C P < 0.001 

Whole blood Se levels and GPx activity in AA cows at end grazing 

season were markedly different between the herds. Herd A showed the 

lowest Se levels and GPx activity, i.e. pronounced Se deficiency. Herd 

B showed higher Se level and GPx activity, but still deficient. Herd C 

showed significantly higher Se and GPx levels, within the 

physiological ranges. Differences in body condition between the herds 

were not observed, but 2 cows died in Herd A. Herds B and C had no 

deaths. Death causes were not identified. There were some differences 

in numbers of pregnant cows after natural mating. 75%, 80% and 95% 

conceived in Herds A, B and C, respectively. 

Conclusion: Organic Se favourably influenced Se levels and GPx 

activity in blood and fertility. Mineral licks with sodium selenite seem 

to be worst source of Se. The mineral licks did not remove Se 

deficiency, which was reflected in very low Se levels and GPx activity 

and worse fertility results. The results support the previous findings that 

the utilization of Se from inorganic sources by cattle is poorer than that 

from organic sources. The differences between Herds A and B were also 

due to a lower intake of the lick than tha of the mineral feed (meal). 

The study was carried out within the research project no. 1G46086 

(NAZV). 

Key words: beef cows, organic Se, Se deficiency, metabolic profile 

509 Effects of Various Anti-Stress Additives in Daily Diets on the 

Microelements of the Heat Stress Cows 

H. Yuan, De. He, J. Wu, Li. Wen, Li. Yuan, R. Li, M. Zhang 

Hunan Agricultural University, College of Veterinary Medicine, 

Changsha, Hunan, China 

Twenty healthy and lactating cows with similar weight, corporeity, 

appearance, and productive capability, were selected from the cattle farm 

of Hunan Agricultural University, in the hot season on August 1-30, 2007. 

These cows were classified into 4 groups using a method of single-factor 

comparison in order to evaluate the effects of various anti-stress additives 

on the microelement in blood, milk, and hair of the heat stress cows. The 

results showed that the concentration of zinc in blood was significantly

increased in the cows fed with the diets supplemented with vitamin C at a 

dosage of 1100-1500 mg/day compared with the control group (P

and HCO 3 level. The concentration of K ions holds in reference values. 

The concentration of Na ions was various, but still was on proper 

levels. 

Conclusions: The DCAB levels of diets oscillated between +66 and 

+228 meq/kg DM and were lower then optimal. Obtained BE indices 

were higher than reference value, that stated about alkalosis 

occurrence. 

Key words: dietary cation-anion balance, feeds, dairy cows 

514 GH Serum Values in Dairy Cows with Clinical Primary 

Ketosis 

A. Cuomo 1 , L. Scaletta 1 , G. Bertoni 2 , L. Esposito 3 

1 

Faculty of Veterinary Medicine, Dipartimento di Scienze Cliniche 

Veterinarie, Teramo, Italy 

2 

University Cattolica Sacro Cuore, Istituto di Zootecnica, Piacenza, 

Italy 

3 

Faculty of Veterinary Medicine, Dip. Scienze Zootecniche ed Isp. 

Alimenti, Napoli, Italy 

The objective of the present study was to investigate the GH values in 

dairy cows with spontaneous clinical ketosis. Eight ketotic Holstein 

Fresian cows (KT group) within the sixty days after parturition, with ßhydroxybutyrate 

(BHBA) values higher than 1.2 mmol/L and 

nonesterified fatty acid higher than 0.6 mmol/L, were evaluated for 

metabolic profile, plus RIA for insulin and GH. Before sampling the 

clinical diagnosis of ketosis was confirmed by portable dry analyzer 

Medisense-Optium Abbott. A control group of six normal no lactating 

cows (CT group) was evaluated in the same way. Any subject was treated 

by drugs before sampling and the samples were collected at 09.00 a.m. in 

both groups. Statistical analysis was performed respectively in the 

groups and between the groups, by software GraphPad Prism 5. In the 

group KT mean value was respectively for GH 2.473 ab ng/ml (SD 

1.038), insulinemia, 4.570 bcA µU/ml (SD 2.101), glycaemia 46.00 bC 

mg/dl and (SD 7.746), BHBA 2.213 ab mmol/L (SD1.606) and NEFA 

1.796 ac mmol/L (SD 0.5299). In the CT group mean value was 

respectively for GH 2.162 a ng/ml (SD 0.9184) insulinemia 8.445 bB 

µU/ml (SD 1.082) glycaemia 71.50 ABC mg/dl (SD 8.264) BHBA 

0.283333 a mmol/L (SD 0.075277) and NEFA 0.1904 a µmol/L (SD 

0.1692). All the results are in accordance with the literature except the 

GH in KT group. Usually in cow 60 days post-parturition the GH value 

ranges around 5-10 ng/ml, significantly different respect our data in 

ketotic cows for the same period of lactation. Statistical analysis shows 

between the groups significance for insulinemia, glycaemia, BHBA and 

NEFA values as expected, at difference of GH that is not significant. No 

correlation has been observed between NEFA and GH values in the KT 

group, at difference of positive correlation described in literature for 

ketosis-free lactating cow. We hypothesize, that low GH concentration in 

dairy cow affected by primary ketosis, could have a protective effects 

against more pronounced fats mobilization during NEB. 

Key words: cow, ketosis, GH 

515 Romifidine as Treatment for Primitive Ketosis in Dairy Cow 

A. Cuomo 1 , L. Scaletta 1 , G. Bertoni 2 , L. Esposito 3 

1 

Faculty of Veterinary Medicine, Scienze Cliniche Veterinarie, 

Teramo, Italy 

2 

University Cattolica Sacro Cuore, Istituto di Zootecnica, Piacenza, 

Italy 

3 

Faculty of Veterinary Medicine, Dip. Scienze Zootecniche ed Isp. 

Alimenti, Napoli, Italy 

Romifidine (RMF) is an a-2 agonist utilized like analgesic-sedative drug 

in dog and horse, and rarely in cow. The a-2 agonist drugs induce a 

transient anabolic effect with hyperglycaemia, biphasic hypohyperinsulinemia 

and a rise of GH concentration. The objective of this 

work was to evaluate the effectiveness of Romifidine in the treatment of 

ketosis and its metabolic effect in dairy cows. Eight dairy cows affected 

by clinical ketosis (KT group), with ß-hydroxybutyrate (BHBA) values 

higher than 1,2 mmol/L and nonesterified fatty acid (NEFA) higher than 

0.6 mmol/L, were included in the group. All the animal displayed the 

syndrome in the sixty days after parturition and do not received treatment 

before our study. Six not lactating cows with normal BHBA values were 

enrolled in the control group (CT). Both groups were evaluated, for 

metabolic profile plus GH and insulin, to study the RMF effects in a 

negative energy balance mode (NEB) like group KT and in a positive 

energy balance (PEB) like group CT. In KT group RMF was injected at 


09.00 a.m. for 5 days at dose of 7.5 Ïµg/Kg/i.m, after collection of basal 

blood sample and evaluation of levels of BHBA, by portable dry analyzer 

Medisense-Optum Abbott. Adjunctive sample was collected at day 1 after 

2.5 hours from RMF injection. In CT group the samples were collected 

before injection and at 1; 1.5; 2; 2.5;3; 24 hours after it to evaluate a 

complete daily curve. Each sample was processed for metabolic profile 

plus insulin and GH and data were statistically processed. The sedative 

RMF at 7.5 µg/Kg/i.m. induces a significant increase in GH and 

glycaemia values and a biphasic hypo-hyperinsulinemia in both groups, 

without any sedation effect. GH mean value shows significant differences 

between KT and CT groups, before and after 2.5 hour RMF injection 

respectively of 2.473 ab (SD 1,038) and 9.225 A ng/ml (SD 8.774) in KT 

and 2.162 a (SD 0.9184) and 3.022 a ng/ml (SD 1.206). In the group KT 

BHBA mean value falls down at 1 mmol/L (SD 0.7838) and food intake 

normalizes within 5 days. RMF is effective as unique treatment for 

primitive ketosis in dairy cow. We hypothesize, to explain this action, a 

role of described hyperglycaemia and transient hypo-hyperinsulinemia, 

plus the normalization of GH values, without the predictable increase 

values of NEFA, wich is typical for GH action in NEB cow. For this 

reason we could hypothesize some additionally, direct actions of RMF not 

correlated with GH activity. 

Key words: cow, ketosis, G˙, romifidine 

516 Mycotoxin Screening in Healthy Cows and Cows with 

Abomasal Displacement 

A. Alkaassem 1 , M. Fürll 1 , S. Dänicke 2 

1 University of Leipzig, Medizinische Tierklinik, Leipzig, Germany 

2 Federal Agricultural Research Center (FAL), Animal Nutrition, 

Braunschweig, Germany 

Objectives: The aim of this study was a) to examine samples of fodder, 

blood, milk and bile for concentration of deoxynivalenol (DON), 

Zearalenone (ZON) and their metabolites de-epoxy-DON, a-zearalenol 

(ZOL), ß-ZOL, zearalanon (ZAN), a-Zearalanol (ZAL) as well as ß- 

ZAL, b) to study possible relations between the toxin arrears and 

diseases. 

Material and methods: We checked 61 ill cows with abomasal 

displacement and accompanying diseases as well as 13 healthy HF 

cows. 

Results: 77.8% Patients without mycotoxins were cured. From those 

with mycotoxins 18 (72%) were cured without clinical reactions; 7 had 

to be euthanized due to severe accompanying diseases and severe 

haematological or clinical chemical deviations in the blood being 

typical for abomasal displacement. We do not observed specific 

clinical mycotoxins effects on health. ZON/ZOL associated changes in 

the ovaries and uterus could not be macroscopically determined. The 

transabdominale collection of bile by aspiration can be easily 

performed in sick cows. In healthy cows mycotoxins were not found. In 

the slightly contaminated fodder samples the mycotoxin concentration 

were 0.161 g DON /kg (0.086-0.191) and 6.35 µg/ ZON kg (4.88-7.85). 

None milk samples contained mycotoxins. DON (0,002 µg/ml) was 

found in one of 61 serum samples, de-epoxy-DON in four samples 

(8%). In bile de-epoxy-DON (37.6 µg/ml) was found only once. 39% of 

the examined bile samples were contaminated with ZON and/or its 

metabolites (ZON 9.85 ng/g [8.10-16.33], b-ZOL: 59.9 ng/g [5-78] and 

ß-ZOL: 37.6 ng/g. In all cases concentrations were within the 

physiological range. The metabolic parameters protein, albumin, urea, 

kreatinin and TEAC did not show differences in cases with or without 

mycotoxins. Mg, Ca, Na, K, Cl, pH, PCO2, bilirubin, glucose, FFA and 

cholesterol concentrations were not in physiological ranges in all cases 

independed on mycotoxins. The haemogram of all patients with or 

without mycotoxin did not show deviations. AP, GGT and CK 

activities of all patients were within the normal ranges. However mean 

AST and GLDH activities of all mycotoxin positive patients were 

slightly increased compared to mycotoxin negative cows. 

Conclusions: We concluded that no specific clinical, haematology or 

clinical chemical changes for mycotoxin influences (DON, ZON) could 

be determined. 

517 Strong Electrolytes (Sodium, Potassium, Chloride) Determine 

Hydrogen Ion and Bicarbonate Concentrations in Urine of 

Ruminants 

H. Staempfli 1 , P. Constable 2 

1 

Ontario Veterinary College, Department of Clinical Studies, 

Guelph, Canada

2 Purdue University, Department of Veterinary Clinical Sciences, 

West Lafayette, United States 

Urine contains 5 strong ions (high Na, K, Cl, and low Ca, Mg) and 4 

buffers over the physiologic range for urine pH (2 volatile buffers 

[bicarbonate and ammonia] and 2 non-volatile buffers [phosphate, 

creatinine]. Translating physicochemical approach to urine it is 

hypothesized that the primary determinants of the urinary pH are the 

strong electrolyte concentrations represented by the strong ion 

difference (SID) [(Na +K)- (Cl)] urine. The theoretical polynomial 

relationship between SID vs pH and SID vs HCO 3 in simulated urine 

containing strong ions and normal pCO 2 predicts very high urine 

bicarbonate concentrations (>200 mEq/L). Normal urine samples were 

collected from 5 mature Holstein dry cows and analysed on a multianalyzer 

system (Hitachi 911) for concentrations of Na, K, Cl, and 

TCO 2. PH (pH meas) was measured using an Accumet-benchtop pHmeter. 

The pH (pH calc) was then calculated using SID in the Stewart 8 

model equation on an excel spreadsheet (MS Office 2003) with pCO 2 

kept at 40 mmHg and individual SID from each urine sample (table). 

The high level of bicarbonate concentrations observed in urine 

confirms that bicarbonate is a dependent variable adjusting to 

concentration of SID based on dissociation equilibria and law of 

electroneutrality. Electrolyte intake and renal excretion are very closely 

correlated and alkaline pH in urine of ruminants is strongly associated 

with surplus strong ion intake, especially with diets high in potassium 

concentrations. There was good correlation between pH calc and pH meas 

in this small sample size. Electroneutrality was off, indicating 

measurement problems with ion specific electrodes. In conclusion pH 

and bicarbonate concentration (TCO 2) of urine are primarily dependent 

on SID. 

Strong electrolytes, bicarbonate (TCO 2) and pH in Urine of 5 dry cows 

Na K Cl SID pH calc pH meas TCO 2 E- 

(mEq/L) (mEq/L) (mEq/L) (mEq/L) (mEq/L) Neutrality 

(mEq/L) 

10 388 143 255 8.4 8.5 229 -26 

108 276 110 274 8.4 8.4 227 -47 

129 246 204 171 8.2 8.4 91 -80 

37 306 133 210 8.3 8.3 148 -62 

95 320 178 237 8.4 8.5 218 -19 

Key words: acid-base, urine, strong-ions, electrolytes, pH 

518 Fat Mobilization Syndrome and Blood Coagulation in Cows 

K. Waldau-Ringel 1 , M. Müller 2 , M. Fuerll 1 

1 

University of Leipzig, Department of Internal Medicine, Leipzig, 

Germany 

2 

University of Leipzig, Department of Clinical Diagnostics, Leipzig, 

Germany 

Objectives: During the fat mobilization pro- and anti-inflammatory 

cytokines are released from the adipose tissue. These could cause a 

circulatory insufficiency and the lipolysis is increased by insulin 

inhibition. Dystocia with placental retention and endometritis following 

bacterial determined endotoxin release are caused by enhanced cytokine 

development. Cytokines could activate the coagulation cascade, so that 

hemostasis disorders and a disseminated intravascular coagulopathy are 

found. The aim of the study was to evaluate, if dairy cattle with fat 

mobilization syndrome or endometritis have coagulation disorders in 

dependence of disease at admittance into the clinic. 

Material and methods: Blood was collected from 60 cows, which 

were admitted into the clinic because of abomasal displacement and 

accompanied diseases. From these 60 cows, a basal therapy was given 

to group 1 (n=25) and additional a single dose dexamethasone was 

given to group 2 (n=25). 10 cows died in spite of intensive care. 

Comparative 36 healthy cows were tested. The numbers of 

thrombocytes and leucocytes were analyzed in EDTA blood and a 

number of haemostatic parameters like prothrombin time (PT), 

fibrinogen (FIB), reptilase time (RT), Antithrombin (ATIII), fibrin 

degradation products (D-Dimers), factor XIII (F XIII) and the activated 

protein C (APC) in citrated blood. Additional as acute phase proteins 

haptoglobin and Procalcitonin (PCT) were tested. 

Results: 50 cows could discharge from clinic after 4 days. 6 cows died 

within 24 hours and 4 cows died after 4 days. Fibrinogen was increased 

at first day and decreased while treatment. Antithrombin and factor 

XIII decreased in cows which died faster than in cows which were 

cured. The fibrin degradation products were slightly increased and 

were sunk while treatment. Cows, which died, had highly increased d- 

Dimers. Procalcitonin and haptoglobin were increased in ill cows and 

decreased during treatment. 

Conclusions: Dairy cattle which suffer from fat mobilization 

syndrome or endometritis have a poor prognosis concerning the acute 

phase proteins fibrinogen, haptoglobin and procalcitonin and the 

haemostatic parameters especially antithrombin, factor XIII and d- 

Dimers. A single dose of dexamethasone has no adverse effects to the 

coagulation, but shows a stabilizing effect. An additional 

administration of heparin is indicated. 

Key words: cattle, blood coagulation, fat mobilisation syndrome, 

coagulation disturbances 

519 Oxalates Plants: Reviewed of Eco Toxicological Aspects 

among Farm Animals 

M. Abdallah 1 , T. Abdelkrim 2 , K. Gerard 3 

1 

Centre Universitaire d’El-Tarf, Veterinary Department, El-Tarf, 

Algeria 

2 

Université d’Annaba, Biology Sciences, Annaba, Algeria 

3 

Ecole Vétérinaire, Veterinary Department, Lyon, France 

The poisonings by oxalic acid and organic salts (the oxalates) are 

especially known of the veterinary practitioners as poisoning due to the 

ethylene glycol (antifreeze), which oxidized in oxalic acid in the liver, 

is at the origin, in particular, of interstitial nephritis and deadly 

epithelial among the domestic animals (carnivores). However, some 

plant poisonings to the oxalates are not diagnosed correctly, probably 

by ignorance. They are due to plants rich in oxalates and which 

diagnosis is not so difficult. The aim of our work is to bring a maximum 

of relative information to the eco toxicological aspect of these toxic 

substances composing these plants. The example of the oxalis is given, 

very rich oxalates plant that infests the natural grasslands as those 

sown. The pathological aspects that especially interest the farming 

activity veterinarian more that the canine practice are discussed as well 

as the pathogenic aspect of the poisonings in different animal species, 

especially in bovine species (cattle). 

Key words: oxalates plants, oxalic acid, eco toxicology, pathologies, 

cattle 

520 Saponins and Pithomyces chartarum Spores Evaluation in 

Pastures from Southwestern of Goiás State, Brazil 

K. Brum 2 , MC. Fioravanti 1 , M. Haraguchi 3 , LA. Silva 1 , 

F. Riet-Correa 4 , JR. Borges 5 , J. Souza 1 , E. Campos 1 , M. Pires 1 , 

A. Pinto 1 

1 Universidade Federal de Goiás, Escola de Veterinária, Goiânia, 

Brazil 

2 Universidade Federal do Mato Grosso do Sul, Campo Grande, 

Brazil 

3 Intituto Biológico, Sao Paulo, Brazil 

4 Universidade Federal de Campina Grande, Centro de Saude e 

Tecnologia Rural, PATOS, Brazil 

5 Universidade de Brasília, Faculdade de Veterinária, Brasília, Brazil 

Brachiaria species are important forages in tropical areas. In Brazil, there 

are around 51 millions of hectares that are feed sources to livestock. 

Outbreaks of hepatogenous photosensitization have been reported in 

ruminants grazing B. decumbens and B. brizantha. The disease was 

attributed to the presence of Pithomyces chartarum spores, but recent 

studies suggest that the steroidal saponins contained in the forage could 

initiate photosensitization outbreaks. General objectives of this study 

were verify if the quantity of protodioscin, a steroidal saponin, and the 

spores quantity variation during the year; and to inquire if the saponin 

was present in other forages that was grazing by the animals. With two 

month of interval, six harvests was taken in ten farms located in Jataí, 

Mineiros and Perolândia cities of the southwestern region of Goiás State, 

Brazil. The spore count was made second a standard technique. The 

butanol-soluble residue containing saponin was submitted to thin layer 

chromatography and spectrophotometric analysis using Ehrlich reagent 

in 515nm. P. chartarum spores found in the pastures varied of 0 to 

40.000 spores/g The number of of grass and the biggest amount was 

observed in the July month (dry season in Brazil). Protodioscin was 

isolated only in B. decumbens and B. brizantha leaves; the biggest 


amount of saponins was observed in March, in the rain season, and the 

lowest amount in July. These results shoed the great variation in levels of 

saponins during the experiment, suggesting a relation between the 

pluviometric precipitation and/or cycle of life in these species of 

Brachiaria evaluated. Acknowledgements to Coordenaçao de 

Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and to 

Conselho Nacional de Desenvolvimento Científico e Tecnológico 

(CNPq) by financial support. 

Key words: brachiaria, spectrophotometry, sporidesmin, 

photosensitization, protodioscin 

521 The Effect of Dry Period versus Continuous Lactation on ßcarotene, 

Vitamin A and Vitamin E in the Mammary 

Secretion 

F. Schweigert 1 , A. Wesser 2 , I. Buchholz 2 , J. Raila 1 

1 

University of Potsdam, Department of Physiology and 

Pathophysiology, Potsdam-Rehbruke, Germany 

2 

University of Leipzig, Department of Veterinary Physiology, Leipzig, 

Germany 

ß-carotene and fat-soluble vitamins A and E are accumulated during the 

dry period in the mammary gland and secreted with colostrums together 

with the accumulated immunoglobulins. Both are of great importance for 

the health of the new borne calf. In this study changes in fat-soluble 

vitamins A, E and ß-carotene (rpHPLC) and IgG1 (immunodiffusion) in 

the secretion of the mammary gland during late gestation in dry dairy 

cows (n=12) and those which were milked until parturition (n=5) were 

compared over a total period of 11 weeks starting 8 before parturition. 

Results showed a biphasic increase in all components during the dry 

period. The first approx. 30 d ap and the second closely toward parturition. 

In animals with continuous lactation an increase towards parturition was 

observed as well but maximum levels were reached earlier approx. 20 ap. 

Absolute levels were comparable in both groups. In both groups low 

levels characteristic for mature milk were reached within a few days after 

parturition (P

metabolism. The objective of this study was to evaluate the effect of 

the intramuscular administration of MPSP on blood serum lipid 

variables and selected reproductive parameters in dairy cows. Thirty 

six Holstein cows within the 2 nd and 4 th lactation with body condition 

score between 3.5 and 4.5, 10 to 6 days before expected calving date 

were randomly divided into two groups. Group 1 (n=18) received 

three application of 50 mL (5 g of MPSP) at 20 day-intervals. The 

first application was 10 to 6 days prepartum, the second and the third 

administration were 10 d and 30 d postpartum, respectively. Group 2 

(n=18) receiving no treatment. Blood samples were collected in all 

cows from tail vein on day 10 to 6 before expected parturition date 

and on day 10, 30 and 40 after calving. Serum free fatty acids (FFA), 

b-hydroxybutyrate (BHB), cholesterol (CHOL) and triacylglycerols 

(TG) were measured. The results were analyzed with repeated 

measures analysis of variance. A Chi-square test evaluated the 

proportion of cows with increased concentrations of FFA (>0.4 

mmol/L) and BHB (>1.4 mmol/L). There were no significant 

difference in FFA and BHB between Group 1 and Group 2 (P>0.05). 

Before parturition, BHB concentrations were lower than those after 

calving (P0.4 mmol/L) before calving was higher in 

Group 1 (P1.4 mmol/L) was lower in Group 1 at days 30 and 40 postpartum 

(P

Water was offered ad-libitum. Feed supplements were offered in the 

milking parlor (treatments I and III) and 2-3 hours before milking 

(treatment II). Stocking rate for the 86 days trial were 2.07, 2.08 and 

2.57 cows/ha for treatments I, II and III, respectively. In forage 

samples, in average, concentration (mg/Kg DM) of Lolitrem B 

(0.9±0.1) was higher than Ergovaline (0.5±0.1). Daily milk yield, 

adjusted to 4% fat content, was 12.9 b , 14.5 ab and 15 a L. for treatments 

I, II and III, respectively (P0.05) in the plasma Mg values. CUM values increases 

(p

4-ipomeanol and 1,4 ipomeanol. These compounds when absorbed into 

the animal are further converted by microsomal activity in pneumocytes 

to a toxic lung oedema factor, which is responsible for the pulmonary 

pathology. The increased use of sweet potatoes for human consumption in 

the UK was noted as early as 1975 by Coxon and others. They are now 

grown in Europe and increasingly fed as a by-products to cattle. This is 

believed to be the first such case published in the UK. 

Key words: sweet-potato, toxicity, respiratory, interstitial, pneumonia 

531 Key Hepatic Gluconeogenesis Enzymes Activity in Response 

to Long-term Glucose Infusion in Dairy Cows 

B. Al-Trad 1 , K. Reisberg 1 , T. Wittek 2 , G. Gäbel 1 , M. Fürll 2 , 

J.R. Aschenbach 1 

1 Faculty of Veterinary Medicine, University of Leipzig, Institute of 

Animal Physiology, Leipzig, Germany 

2 Faculty of Veterinary Medicine, University of Leipzig, Medizinische 

Tierklinik, Leipzig, Germany 

Background and Aims: Periparturient ketosis in dairy cows is 

characterized by an imbalance of glucose and lipid metabolism. 

Insufficient gluconeogenesis is thought to be primarily caused by a lack 

of glucogenic precursors due to negative energy balance. However, it is 

not clear whether insufficient catalytic capacity of gluconeogenic 

enzymes might contribute to insufficient glucose production 

postpartum. The purpose of the present study was to investigate (1) 

whether a positive energy balance due to chronic glucose infusion has 

a depressive effect on selected gluconeogenesis enzymes activity in 

dairy cows and (2) how quick such depressive effect might be reversed 

after stopping surplus glucose supply. 

Material and Methods: Twelve midlactation dairy cows were 

maintained over a 28 d experimental period on continuous intravenous 

infusion of either saline (control group, six cows) or 40% glucose 

solutions (treatment group, six cows). The infusion dose was calculated 

as a percentage of daily energy (NEL) requirements, starting at 1.25% 

and increasing by 1.25%/d until a maximum dose of 30% was reached. 

Dose was then maintained at 30% for 5 d. No infusions were made 

between d 29-32. Blood samples were taken every 2 d and liver 

biopsies were taken on d 0, 8, 16, 24, and 32. 

Results: Blood glucose and insulin concentrations showed only slight and 

occasional increases while NEFA and BUN concentrations decreased by 

glucose infusion. Liver glycogen increased progressively during glucose 

infusion until reaching plateau between d 16 and 24. Liver glycogen fell 

back to baseline values on d 32 after cessation of glucose infusion. During 

the infusion period, no significant changes were observed in the activities 

of phosphoenolpyruvate carboxykinase and glucose 6-phosphatase. 

Fructose 1,6-bisphosphatase decreased and pyruvate carboxylase tended 

to decrease in the treatment group during the high dose of glucose infusion 

(i.e. infusion dose of 20% & 30% NEL requirements) but quickly returned 

to baseline after stopping the infusion. 

Conclusion: The activity of key hepatic gluconeogenesis enzymes in 

lactating dairy cows is not significantly affected by long-term increases 

in glucose supply. Excessive glucose improves body tissues energy 

status without impairing the role of liver as the main source of glucose 

production in dairy cows. Thus, dysregulation of gluconeogenic 

enzymes activity by a positive energy balance pre-partum is likely not 

a major cause of postpartum metabolic disturbances. 

532 Milk Drop Syndrome in Dairy Ewes: Clinical and Diagnostic 

Approach 

N. Giadinis 1 , G. Arsenos 1 , G. Fthenakis 2 , P. Tsakos 3 , C. Dovas 1 

1 

School of Veterinary Medicine, Aristotle University, Thessaloniki, 

Greece, Greece 

2 

University of Thessaly, Thessaloniki, Greece 

3 

Institute of Infectious and Parasitic Diseases, Centre of Veterinary 

Institutes of Thessaloniki, Thessaloniki, Greece 

“Milk drop syndrome” in dairy sheep is a problem with considerable 

financial implications. However, it has not been thoroughly investigated 

and research data are scarce. In this paper, we present the results of a 

study, carried out from 2003 to 2007, into the factors involved in milk 

drop syndrome reported in 18 flocks of dairy sheep in Northern Greece. 

The appropriate clinical and diagnostic approach is also discussed. In 

each flock, five adult ewes were selected randomly among those with 

reported milk drop. Samples of blood, milk and faecal samples were 

collected from these animals throughout a milking period. Their feeding 

regime was also evaluated. Evaluation of milk yield was based on data 

of previous milking records. In 10/18 flocks, the subclinical mastitis, 

caused by coagulase negative Staphylococcus spp. or Streptococcus spp, 

was the reason for the reduced milk yield. In other flocks the reasons 

were contagious agalactia confirmed by isolation of Mycoplasma 

agalactiae in milk cultures (1 flock), inappropriate and unbalanced 

nutrition with a ration containing suboptimal protein (1 flock), chronic 

ruminal acidosis (1 flock) or Maedi-Visna confirmed by PCR, serum test 

for antibodies and histological findings in mammary glands (2 flocks). A 

combination of the above factors was observed in 3/18 flocks, while in 

one flock these factors were coupled with intense gastrointestinal 

parasitism. The results confirm that subcilincal mastitis is the major 

reason for reduced milk yield. The role of different pathogens and the 

need for proper management of animals in individual flocks imply that 

the clinical and diagnostic approach should involve a farm investigation 

before any measures are taken. 

Key words: dairy ewes, milk, mastitis 

533 The Relation of ante Partum Backfat Thickness with 

Metabolic Disorders and Fertility Problems in Dairy Cows - a 

Long Term Study 

H. Pothmann-Reichl 1 , F. Zimmer 2 , I. Sommerfeld-Stur 3 , C. Iben 4 

1 University of Veterinary Medicine Vienna, Teaching and Research 

Farm, Vienna, Austria 

2 Veterinary Clinic, Private Veterinary Practice, Oberkirchen, 

Germany 

3 University of Veterinary Medicine Vienna, Clinical Department of 

Animal Breeding and Reproduction, Vienna, Austria 

4 University of Veterinary Medicine Vienna, Department of Veterinary 

Public Health and Food Sciences, Vienna, Austria 

ObjectivesIn this retrospective study the influence of post partum change 

of back fat thickness (BFT) in cattle on fertility, metabolic problems, 

mastitis and milk yield was investigated. It is hypothesised that a higher 

decrease of back fat thickness post partum (p.p) has a negative impact on 

fertility and fat metabolism. Animals, materials and methodsFive dairy 

farms in Germany (292 cows) and two farms in Austria (111 cows) 

participated in this trial and 1045 lactation periods were analysed. The 

back fat thickness was measured with an ultrasound (3.5 MHz) between 

the tuber coxae and the tuber ischiadicum at the following days: 8 weeks 

ante partum (a.p.), 42 days a.p., day of parturition, 42 days and 100 days 

p.p. The programme “Inter-Herd”(Interagri, Earley Gate, UK) was used to 

record the data. Statistic evaluation was done by SPSS for Windows 

14.0.1 using odds ratio (OR >1 increased risk, OR

Se, I and Co) more than 500 blood samples were analyzed from 70 

herds, several of them with problems of infertility, abortions, 

premature calving, weak calves, enzootic haematuria and high somatic 

cell counts. The majority (91%) of the analysis were performed in 

Holstein-Friesian (HF) cattle, of which 284 were milking cows (MC), 

129 heifers (H), 13 calves (C), 43 bullfight cattle (BF) and 12 beef 

cattle (B). The results of the TE survey indicate that 92% of the animals 

had both low levels of plasmatic iodine and thyroxin (T4) 84% of the 

herds had at least one animal with Co deficiency (75% BF cattle, 54% 

B cattle, 51% H and 37% MC). About 31% of the MC had low levels 

of Se (glutathione peroxidase, GSH-Px1) and serum Cu, as well as 38% 

of serum Zn. The most severe deficiencies were identified in the heifers 

group. To complement these data, 7 clinical trials were developed 

using HF reared on pasture grazing systems. In each trial there were 

two randomly constituted groups: a control group and a group have 

given extra feed supplements. The individual weights of animals were 

measured every 14 days. Blood samples were taken at the beginning 

and end of the trials for TE levels and blood cell counts. Different 

supplementation protocols and products were implemented in each 

trial. The clinical trial results indicate that TE supplemented yearlings 

had significantly higher productive output and hematological 

parameters than the controls. The least laborious, most consistent and 

reliable supplementation system studied, was obtained with the intraruminal 

slow releasing bolus (SRB) from Alltrace, Agrimin (UK). 

However, similar results were obtained when Se and I were added to 

the drinking water. A TE survey should be included in the sanitary and 

management plans for each herd in order to enhance animal 

performance and health, as well as farm profitability. 

Key words: cattle, trace elements, mineral deficiency, azores 

535 Influence of Postpartum Propylene Glycol Supplementation 

on Metabolism, Animal Health, Reproductive Performance 

and Production in Dairy Cows 

M. Iwersen, U. Falkenberg, R. Voigtsberger, W. Padberg, 

A. Mertin, W. Heuwieser 

Freie Universität Berlin, Tierklinik für Fortpflanzung, Berlin, 

Germany 

The effect of postpartum (pp) supplementation of 250g propylene 

glycol (Propy) per cow and day via a total mixed ration (TMR) on 

metabolism, animal health, reproductive performance and milk 

production was studied on a commercial dairy farm in Germany. 

A total of 258 multiparous Holstein-Friesian cows were enrolled in a 

randomized and controlled field study. On the basis of expected calving 

date and previous milk production animals were blocked and allocated 

to one of the two treatment groups. The TMR fed in group 1 (Control, 

n=131) was supplemented with Propy before offering it to group 2 

(Study, n=127). The TMR was mixed and offered twice daily for a 

period of 40 days pp. Blood samples were collected within 12 hours at 

the day of parturition, on day four, ten, twenty and forty. Samples were 

centrifuged and serum was stored at 20 °C until analysis. Body 

Condition Score (BCS) and Back Fat Measurement (BFT) values were 

recorded on the same days. Serum samples were shipped to one 

commercial laboratory for the determination of Nonesterified fatty 

acids (NEFA), Betahydroxybutyrate (BHBA), Triglycerides, Ketone 

bodies, Bilirubin, Glucose and activities of Aspartate-aminotransferase 

(AST), Glutamate-dehydrogenase (GLDH), Alanine-aminotansferase 

(ALT) and g-glutamyl-transferase (GGT). Metabolic parameters were 

analysed by a 6-step UNIANOVA with repeated measurements using 

SPSS ® -Software. ALT, GGT, GLDH, Insulin, Bilirubin and Glucose 

were comparable between the two groups. Concentrations of AST, 

Triglycerides, BHBA and Ketone bodies were significantly (p

had greater trans11-18:1 percentage in milk fat (2.39%) this response 

resulted in lower milk fat (3.47%). In the farm C the percentage of CLA, 

the ratio of trans11-18:1 to CLA and the ratio (6:3) of FA were very 

beneficial according to human health demands. Milk fat content was 

negatively correlated to trans11-18:1 percentage (r=-0.93) and to CLA in 

the milk fat (r=-0.85). Our data suggest that rate of ruminal degradation of 

NDF influences the milk fat content. Results showed that there was a 

negative correlation between milk fat content and degradation rate of 

NDF (r=-0.91). It can be assumed, that even suboptimal fiber levels in 

TMR containing slower degradable fiber may be sufficient to maintain 

proper rumen fermentation and level of milk fat synthesis. This finding 

suggests that in the TMR of cows not only the quantity of fiber fractions 

(NDF, ADF) has to be determined, but even their degradation rate should 

be taken into consideration. 

538 The Effect of Antioxidant and Omega-3 fatty acid 

Supplement on Metabolic Status and Reproductive 

Performance in Postpartum Dairy Cows 

F. Tóth 1 , Gy. Gábor 1 , O. Balogh 1 , G. Koppány 2 , H. Fébel 1 

1 

Research Institute for Animal Breeding and Nutrition, Herceghalom, 

Hungary 

2 

Vitafort Ltd., Dabas, Hungary 

The objective of this study was to investigate the effect of different 

supplements on metabolic status, the onset of cyclic ovarian and the 

rate of re-conception function in the postpartum (pp.) period. The study 

was carried out on 66 pp. Holstein cows. The cows were fed the same 

total mixed ration in dry period and 5-7 pp. days were divided into the 

control (C) (n=16) and the experimental (E) group (n=50). The feed of 

E was supplemented with extra beta carotene, vitamin E, inactivated 

yeast, selenium and microelement chelates until 50 pp. d.; from 50 pp. 

days onwards the supplement was only 200 g grinded linseed (W3 fatty 

acid supplement). The average milk yield of E cows was 3 kg/day 

higher than C cows from 8 th to 120 th lactation day. The plasma level of 

BHB was higher in the E group. While both groups showed elevated 

NEFA level on day 30, only C group had increased NEFA 

concentration on day 50. Plasma concentration of urea was lower in the 

E group on day 90 and 120 pp. Serum level of carotene in the E group 

was higher during the whole experiment. P4 concentrations between 

Days 18 and 39 after AI were greater for cows fed linseed 

supplementation than the control cows. According to the P4 profile 

more than 80 % of the E group had cyclic ovarian activity versus only 

62.5% of the C group. Although, the conception at first AI (41.2 vs. 

57.1 %) and the overall proportion pregnant (47.1 vs. 71.4 %) was 

lower in the E group. The hypothesis that feeding linseed would 

increase conception rate of dairy cows was not supported by the present 

experiment. The higher P4 level of E cows means the positive effect of 

omega 3 fatty acids and may also have contributed to decrease embryo 

mortality. The applied supplements effected elevated milk production, 

but this higher production led to a negative energy balance (NEB) 

which is confirmed by elevated levels of NEFA and BHB. This 

negative effect of NEB is presumably connected to lower fertility 

performance. 

This work was supported by the project of OMFB-0121 

Key words: conception rate, omega-3 fatty acid, antioxidant, 

metabolic status 

539 Use of hypertonic and isotonic saline solution and furosemide 

for the treatment of the ammonia poisoning in cattle 

S. Kitamura 1,2,3 , A. Antonelli 1 , R. Barreto Júnior 1 , C. Mori 1 , 

M. Sucupira 1 , B. Caputi 1 , E. Ortolani 1 

1 

Faculdade de Medicina Veterinaria da Universidade de Sao Paulo, 

Clinica Medica, Sao Paulo, Brazil 

2 

Universidade Anhembi Morumbi, Clinica Medica, Sao Paulo, Brazil 

3 

Universidade Paulista - UNIP, Instituto de Ciencias da Saude, Sao 

Paulo, Brazil 

To test the efficiency of some treatments for ammonia poisoning in 

cattle 25 steers, never fed non-protein nitrogen before, were used. 

Ammonium chloride solution was infused intravenously (iv) only 

once in each steer until the onset of convulsive episode, and then the 

infusion was immediately stopped. Thereafter, the animals were 

randomly distributed in one of the five different groups, as follows: 

1) control (infusion 300 mL isotonic saline solution - ISS); 2) 

infusion (iv) of ISS 30 mL/kg BW throughout the following 4 h and 

administration by stomach tube of 4 L water. (W); 3) same as group 

2, plus a single dose 2 mg furosemide/kg BW (iv) (F); 4) injection of 

5 mL/kg BW hypertonic saline solution (HSS) (7.2 %) (iv) 

throughout the first 30 min, followed by 20 mL/ kg BW of ISS and 4 

L W.; 5) Same as group 4 and a single dose of F. For the next 4 h 

after the convulsion several clinical variables and the blood 

concentration of ammonia, glucose, lactate, urea, creatinine, sodium 

and potassium, hemogasometric profile, urine global volume and 

urinary excretion of ammonium, urea and creatinine were followed. 

The clinical picture seen just before the treatment was classical and 

caused a characteristic systemic metabolic acidosis, compensated by 

reduction in the pCO 2, hyperlactemia and hyperglycemia. At the 

convulsion the ammonia blood levels were high and similar among 

the groups, but at the 120 th min the animals treated with HSS had 

lower values than the control. The HSS infusion caused, during the 

first 30 min. a marked reduction in the packet cell volume and the 

heart rate. On the other hand, furosemide provoked an increase in the 

heart rate and neither prevented the onset of lung edema, nor an 

increase in the total excretion of urine. Cattle treated with HSS stood 

up and had the return of the rumen movements earlier than other 

groups. The therapy with HSS still increased the urinary volume and 

the total urinary excretion of urea and ammonium during the critical 

1 st h of treatment, while the use of ISS had intermediate results. The 

efficiency of the treatment with HSS or ISS as seen by some 

variables of clinical improvement was better than the control group. 

The higher the urinary urea excretion was, the greater the ammonia 

elimination became. Although with lower efficiency as seen in the 

treatment with HSS, the ISS caused an improvement in the clinical 

picture and at the end of the experiment generated an adequate 

ammonia detoxification. 

Key words: ammonia intoxication (treatment), hypertonic, 

furosemide, cattle 

540 Environmental Low Phosphorus Induced the Erythrocytic 

Membrane Damage: a Pathogenesis of Endemic 

Haemoglobinuria in Dairy Cattle 

JL. Li, Q. Si, FQ. Shi, SW. Xu 


Harbin, China 

Objectives: Dairy cattle could display haemoglobinuria when 

phosphorus was deficient in food, especially in environmental low 

phosphorus in some regions. Endemic haemoglobinuria in dairy cattle 

usually had been detected in some regions of Heilongjiang province in 

china. However, the reason of the endemic haemoglobinuria remians 

unclear. The aim of this study is to illustrate the pathogenesis of 

endemic haemoglobinuria. 

Materials and Methods: In this study, we tested the content of 

phosphorus in soil-grass and serum of 15 cows with obvious clinical 

symptom in Daqing and Anda regions of Heilongjiang province in 

April-July and October, respectively. In addition, we also detected 

Lipid composition, fluidity, viscosity and activity of ATPase of 

erythrocytic membrane of the 15 cows as well as osmotic fragility of 

the erythrocytes and blood routine. 

Results: Our results indicated that the content of soil and grass 

phosphorus was significantly lower (P

541 Variability of Blood Profile and Milk Composition in Relation 

to Stage of Lactation and Season of the Year in Dairy Cows 

SM. Abdel raheem, C. Iben 

University of Veterinary Medicine Vienna, Nutrition, Vienna, Austria 

Objectives of study: The study was designed to determine the seasonal 

and lactational influences on blood metabolites, milk composition, 

BCS, and back fat thickness (BFT) in dairy cows. 

Materials and methods: The study was conducted from April 2006 

until February 2007 using (n=35) dairy cows (27 Fleckvieh and 8 

Holstein Friesian). Cattle were given 15 kg corn silage, 20 kg grass 

silage, and 3 kg grass hay as daily basal feed. A concentrate diet was 

given on the basis of the amount of milk production. Blood samples, 

Milk constituents, BCS and BFT were investigated every 2 months. 

Depending on the day postpartum of sampling, data were divided into 

3 lactation stages; early (up to 75 days postpartum), mid (76 to 150 

days), and late (after 151 days plus dry cows) lactation. Also, 

depending on the season at the time of sampling; the data were divided 

into 4 seasons, spring (April), summer (June and August), autumn 

(October) and winter (December and February). Statistical analyses 

were conducted with the SPSS; ANOVA was used for lactation stage 

and season comparison. The interaction between the two factors of the 

year was done by MANOVA. 

Results: Lactation stage had a significant influence on both blood 

metabolites (glucose, BHBA, blood urea nitrogen (BUN), albumen, 

cholesterol, Mg) and milk components ( milk fat, milk protein milk 

urea) in addition to BCS and BFT. No significant effect of the lactation 

stage was detected on total protein (TP), Ca, P, Cu and Zn. At the same 

time the season of the year has a significant impact on all parameters 

studied except BHBA, P and BFT. BCS was higher at late lactation 

(mean±SD) (3.36±0.41) than early (3.23±0.44) and mid (2.91±0.39) 

lactation. A significant interaction between stage of lactation and 

season of the year was detected for TP, cholesterol and milk protein. 

BFT correlated positively with BCS (r=0.84) and also milk urea and 

BUN (r=0.84) correlated positively. Milk yield and milk protein were 

correlated negatively(r=-0.61). Both BCS and BFT correlated 

negatively with cholesterol r= (-0.4). 

Conclusion: Stage of lactation and season influenced the biochemical 

parameters and metabolites in blood and milk as well as body 

condition. Therefore, the interaction between these two factors be taken 

into account when assessing the nutritional status of dairy cows in 

different climatic periods and in different stage of lactation. 

542 Occurrence of Subclinical Metabolic Disorders and Possibility 

of the Prevention by Licking Block Supplementation in Beef 

Cows 

L. Könyves 1 , V. Jurkovich 1 , A. Tirián 1 , L. Tegzes 1 , O. Szenci 2 , 

Z. Szelényi 2 , E. Brydl 1 



Hungary 


Large Animals, Budapest, Hungary 

The aim of the study was to reveal the occurrence of subclinical 

metabolic disorders in charolais suckler herd in the winter calving 

period and spring breeding season. The biological effect of an 

experimental licking block containing protein source, molasses, 

minerals and vitamins was also evaluated as a possible tool of 

preventing metabolic disorders. Charolais cows (n=114) were selected 

randomly into experimental and control group (n=57-57). From these 

groups 12-12 cows were assigned randomly for metabolic profile tests. 

The experimental group was supplemented with experimental licking 

block (voluntary intake about 1.5 kg/cow/day) from 1 month before 

expected calving until the end of breeding season. Different 

manifestations of energy imbalance (increased fat mobilisation, 

hypoglicaemia, hyperketonaemia) and acid load was diagnosed with 

high occurrence in both groups. Protein-, phosphorous-, sodium- and 

carotene shortage was diagnosed in the control group. The urea, 

phosphorous, carotene concentration in blood plasma and sodium 

concentration in urine were significantly higher in experimental group 

compared to control. The licking block had no effect on energy 

metabolism, acid-base balance, Ca, Mg, K, Cu, Zn, Se and Mn supply. 

There was no difference between groups in mortality- and culling rate, 

the postnatal calves mortality, metritis and the number of veterinary 


treatments. There was 7.3 % higher conception rate in the experimental 

group but the detected subclinical metabolic disorders might play a 

remarkable role in poor reproductive performance in both group. 

Key words: metabolic disorders, beef cow, supplementation 

543 Body Condition Score at Calving and Supplementation with 

Yeast Affect Blood Mineral and Liver Enzyme 

Concentrations in Peripheral Blood of Peri-parturient Dairy 

Cows 

L. O’Grady, R. Alibrahim, V. Gath, M. Doherty, F. Mulligan 

University College Dublin, School of Agriculture, Food Science and 

Veterinary Medicine, Dublin, Ireland 

The objective of this study was to determine the effect of body condition 

score (BCS) at calving and supplementation with a live yeast culture 

(Yea-Sacc 1026 , Saccharomyces cerevisiae, CBS 493.94) beginning two 

weeks pre-calving on the metabolic status of peri-parturient dairy cows. 

Forty Holstein/Friesian cows of mixed parity were randomly allocated to 

two groups depending on BCS approximately 60 days prior to parturition. 

Both groups were fed grass silage ad-libitum for the entire 60 days precalving 

and half of each group were supplemented with a dairy compound 

containing 2.5g of thermo-stable yeast supplement for the last 14 days 

pre-calving and during the first 70 days of lactation. Blood samples were 

taken via jugular venipuncture on the day of calving (12-24 hrs post 

calving) and on days 5, 15, 25 and 35 post-calving and analysed for Ca, 

Mg, P, gammaglutamate transferase (GGT), glutamate dehydrogenase 

(GLDH) and bilirubin (BIL). Data was analysed using ANOVA and the 

Proc GLM statement of SAS separately for each day of sampling as a 2 X 

2 factorial with BCS group, supplement group, BCS group x supplement 

group and parity included in the model. Dairy cows in the high BCS group 

had a higher (P

serum concentrations of Ca, P and Mg were determined by Randox kits 

in semiautomatic biochemical analyzer (Selectra Junior), the 

concentration of 25-OH D 3 by chemiluminescence assay (DiaSorin, 

LIAISON). The concentrations of Ca, P, Mg and 25-OHD 3 were 

compared by repeated measures ANOVA. BCS and BFT were 

compared by student´s t-test. Significant difference was found in 

concentration of Ca, P, Mg and 25-OH D 3 with respect to time sampling 

(p

evaluated, highly significantly greater milk protein was detected in 

those animals ovulating before day 22 p.p. 

Our study demonstrates that these indicators could be useful in 

estimation of the post partal onset of ovarian activity in dairy cows. A 

larger complex of metabolic status indicators in milk will be the subject 

of our subsequent studies. 

This work was supported by a grant No. MSM 6046070901 and 

1G46086 

548 Effect of Organic and Inorganic Zinc Supplementation on the 

Prevention of Zinc Deficiency in Aberdeen Angus Calves 

S. Dresler, L. Perlikova, D. Kumprechtova, J. Illek 

University of Veterinary and Pharmaceutical Sciences Brno, Brno, 

Czech Republic 

Objective: The objective of the study was to investigate the effect of 

supplementation of suckling Aberdeen Angus (AA) calves with 

different forms of zinc during the grazing season. 

Material and methods: At the beginning of the grazing season, zinc 

deficient was diagnosed in AA calves (n=16) at 2 to 5 weeks of 

age along with their mothers, into two groups (Control and Bioplex). 

The calves suckled milk, were given starter feed and gradually were 

getting used to graze. The calf starter feed consisted of crimped wheat, 

oats, barley and protein concentrate. The starter feed contained 36 mg 

zinc per 1 kg and was given ad libitum to calves from 1 week of age. 

After the beginning of grazing season, the zinc concentration in starter 

7 Health Status of Cows Before Parturition Associated with 

Concentration of GM1 & IgG in Colostrum and Reflects 

Resistance of Acquiring New IMI at Early Lactation 

G. Leitner 1 , O. Krifucks 1 , S. Jacoby 2 , Y. Lavi 3 , N. Silanikove 2 

1 Kimron Veterinary Institute, Bacteriology, Bet Dagan, Israel 

2 Agricultural Research Ortganization, the Volcani Center, Animal 

Science, Bet Dagan, Israel 

3 Hebrew University, Rehovot, Israel 

Objective: Study the association between the concentrations of IgG 

and Gangliosides type M1 (GM1) in colostrum of cows and IMI during 

first weeks postpartum. 

Materials and methods: Israeli-Holstein cows n=108: 38 heifers and 

75 pluriparous cows. Cows were dried 60 d before next expected 

parturition. During the 45 d prior to the end of lactation, the cows were 

tested 3 times on the quarter level for bacteriology and SCC. Colostrum 

of cows and heifers were tested on the day of parturition and thereafter 

milk was tested at 10, 30 and 45 d postpartum for bacteriology and 

SCC. Bacterial identification and SCC were carried out according to 

IDF. Total IgG & GM1 (Relative values of GM1, GM1-RV) were 

determined as the reduction of binding of CT to PolySorb plates coated 

with GM1 by competition with the b-chain of cholera toxin (b-CT) 

were determined in the mixed colostrum. The effects of udder infection 

and lactation (heifers vs pluriparous) on total IgG, GM1-RV & SCC 

were determined by a two-way ANOVA in a random design. Linear 

discrimination analysis was used to distinguish between the 

classification variable (uninfected or infected) based on total IgG and 

GM1-RV levels and records of infection. 

Results: High levels of IgG & GM1-RV in pluriparous cows were 

associated with lower new incidence of IMI in the subsequent 

lactation. The mean IgG level in colostrum of heifers was not 

significantly lower than that of pluriparous cows. Infected quarters 

of heifers or cows had significantly lower IgG levels than uninfected 

quarters. The mean GM1-RV in the colostrum of infected heifers 

was significantly higher than that of the pluriparous cows. This 

difference was not found in uninfected animals. Moreover, in 

heifers, GM1-RV in infected quarters was not significantly higher 

than in uninfected ones whereas in the pluriparous cows, GM1-RV 

levels were significantly lower in the infected than in the uninfected 

quarters. Linear discriminate analysis based on these two variables 

yielded 70% successful classification. This analysis was less 

successful in heifers because a high proportion of them joined the 

herd already infected with bacteria. 


Mastitis / Udder Health and Milk Quality 

feed was increased by 30 mg per kg (total 60 mg Zn/kg). The Control 

calves received starter feed with zinc sulphate, the Bioplex calves 

received calf starter with organic zinc (Bioplex-Zn). Calves in both 

groups received starter feed ad libitum. The experiment lasted 2 

months. At the beginning and the end of the trial the calves were 

weighed and each calf was withdrawn a blood sample to measure zinc 

concentration and selected parameters of metabolic profile. The results 

were statistically analysed and evaluated. 

Results: Initial serum zinc concentration was 6.55±0.80 and 7.10±0.6 

umol/l, serum total protein levels were 65.2 l and 66.5 g/l, and serum 

IgG levels were 14.8 g/l and 13.1 g/l in the Controls and Bioplex, 

respectively. At the end of the trial there was a significant increse in 

serum zinc concentrations in both groups, with Bioplex supplemented 

calves showing a higher increase in plasma zinc levels. Zinc 

concentrations were 11.87±1.77 and 15.81±0.72 µmol/l in the Control 

and Bioplex groups, respectively. The differences were statistically 

significant. The Bioplex supplemented calves showed higher IgG 

levels. There were significant differences in daily weight gain between 

the groups, too. While the Control calves showed the growth rate of 

1.12±0.07 kg per calf per day, the Bioplex calves grew 1.24±0.09 per 

calf per day. 

Conclusions: The dietary zinc supplementation in calves favourably 

influenced serum zinc concentrations. The organic zinc 

supplementation resulted in a greater increase in serum zinc 

concentrations, serum IgG levels and weight gain. The organic form of 

zinc can be recommended as a useful tool to prevent zinc deficiency in 

calves. 

Conclusions: The results suggest that the health status of the animal at 

the last month before parturition is associated with colostrum quality 

and reflects its resistance of acquiring new IMI. While in heifers, of 

which high proportion joined the herd when already infected with 

bacteria no such association was found. 

8 Cell Differentiation in Milk of Cows with Healthy Udders 

D. Schwarz 1 , K. Failing 2 , W. Wolter 3 , CP. Czerny 1 

1 Georg-August-Universität Göttingen, Institute of Veterinary 

Medicine, Göttingen, Germany 

2 Justus-Liebig-Universität Gießen, Unit for Biomathematics and 

Data Processing, Gießen, Germany 

3 State Council Gießen, Milk Control, Wetzlar, Germany 

Cell counts and the determination of differential cell patterns in milk 

samples are important criterions for the evaluation of udder health. The 

limits for the evaluation of udder health are well-defined by clear cut-off 

rates. The objective of our study was the analysis of somatic cell 

populations in foremilk samples to identify inflammations at a 

significantly lower level than given by current definitions. The milk of 

Holstein-Friesian cows (n=18) and German Simmental cows (n=2) with 

low cell counts from five Hessian dairy farms was examined 

cytologically. Cell counts were determined with a Fossomatic 5000 and 

bacteriological examinations. Milk smears were generated for each udder 

quarter and stained according to the method of Pappenheim. To create the 

differential cell patterns, 100 cells per milk smear were counted and 

differentiated by microscopy. The classification into different cell count 

groups of all foremilk samples analyzed in the years 2000-2003 in the 

German federal state Hesse showed the importance of very low (7%) and 

low (50%) cell counts. Moreover, the results of monthly cell counts 

developed on quarter levels allowed to suggest inflammable reactions in 

the cell count array lower than 100,000 cells/ml, because there were clear 

cell count variations visible in a little time period. All 80 udder quarters 

tested in our study were subdivided into groups with very low, low and 

medium cell counts. In each group we found epithelial cells at the 

differential cell patterns of a maximum of 3%. The lymphocytes 

dominated the ‘very low’ group with an amount of 70%. The differential 

cell patterns in the ‘low’ group consisted of ca. 40% lymphocytes and 

40% macrophages. The percentage of the polymorphonuclear neutrophils 

clearly increased at cell counts of 43.000 cells/ml. These data indicated 

inflammable reactions at such low cell counts. Our findings were 

confirmed by monitoring the development of the monthly cell counts. The 

bacteriological examinations only identified S. aureus in one quarter, E.

coli in another quarter and Coagulase-negative staphylococci in two other 

cases. The statistical calculations proved a significant correlation between 

cell counts and differential relationships of the four analysed cell types to 

each other. On the basis of the results of our study we detected 

inflammable reactions in the mammary glands of obviously healthy 

udders already at a level of 43.000 cells/ml. 

Key words: differential cell count, low somatic cell count milk, 

differential cell pattern 

9 Understanding Mastitis Epidemiology on Farm - Determining 

the Importance of the Dry Period in Mastitis Control 

A. Bradley 1,2 , M. Green 3,4 

1 University of Bristol, School of Veterinary Science, Bristol, UK 

2 Quality Milk Management Services Ltd., Wells, United Kingdom 

3 University of Nottingham, Department of Veterinary Medicine, 

Nottingham, United Kingdom 

4 Orchard Veterinary Group, Glastonbury, United Kingdom 

Objectives: Appropriately targeting mastitis controls on farm requires the 

clinician to determine the relative importance of the dry and lactating 

periods. The aim of this study was to investigate novel indices for clinical 

and sub-clinical mastitis, previously described by the authors, to 

categorise farms by the importance of the dry and lactating periods. 

Materials and Methods: Using somatic cell counts (SCC) the importance 

of the dry period was investigated by calculating; the Fresh Calver 

Infection Rate (FCIR - proportion of cows with SCCs >200,000cells/ml at 

the first recording post calving), the New Dry Period Infection Rate 

(DPNEW - proportion of cows with SCCs rising from < to > 200,000 

cells/ml across the dry period), the Dry Period Cure Rate (DPCURE - 

proportion of cows with SCCs falling from > to

we measured sensitivity of the entire assay, including the DNA 

extraction procedure. For this experiment, ten mastitis milk samples 

shown to be negative for the tested targets were randomly chosen, spiked 

with known quantities of the target bacterial colony forming units (CFU) 

and analyzed using the PCR assay. When sensitivity was measured based 

on the dilution series of purified DNA, the analytical detection limit of 

the test was as low as 5.6 bacterial genomes per l. When using mastitis 

milk samples spiked with the bacterial targets, the detection limit was as 

low as 100 CFUs per one millilitre of milk. Both experiments showed 

variation in the sensitivity of the test between the different bacterial 

targets. In conclusion, the results of this study demonstrate that the 

PathoProof Mastitis PCR Assay can detect low quantities of bacterial 

DNA. This presentation also discusses some of the challenges involved 

in sensitivity validation of a PCR test in a controlled manner when using 

raw bovine milk as a sample matrix. Sensitivity of the PathoProof 

Mastitis PCR Assay should next be validated against the golden standard 

method, i.e. bacterial culture. 

43 Investigating the Use of Viscosity Measurements for the 

Quantification of the California Mastitis Test (CMT) in Dairy 

Cattle 

J. Roberts 

University of Cambridge, Department for Veterinary Medicine, 

Cambridge, United Kingdom 

The California Mastitis Test (CMT) is widely used as a qualitative 

cow side test for the detection of subclinical mastitis in dairy cattle. 

The viscosity of the milk-CMT gel reaction has been characterised 

but at present no attempt made to determine whether the reaction can 

assess quantitatively the somatic cell count (SCC) in milk samples. 

This initial investigation looks at the milk and milk-CMT gel reaction 

viscosity for various SCC quarters and characterises the sample 

viscosity for these SCCs. 57 quarter samples were collected 

aseptically from 16 Holstein-Friesian cows in a commercial dairy 

herd within the UK. Viscosity measurements were made using a 

Brookfield R/S Rheometer at constant shear rate for milk alone and 

milk with CMT reagent. Each quarter SCC was measured at a 

commercial approved laboratory. The correlations between average 

viscosity, initial viscosity, final viscosity, and change in viscosity for 

both milk and milk-plus-CMT reagent against SCC are demonstrated. 

The removal of the milk viscosity from the milk-plus-CMT reagent 

viscosity may allow quantification of the SCC between animals. The 

milk-CMT gel reaction used qualitatively to demonstrate subclinical 

infection may be able to be measured quantitatively if the process of 

analysing viscosity is carefully controlled as shown by this initial 

analysis. Further work will investigate the CMT reaction at various 

constant shear rates and using controlled shear rates. It is anticipated 

that quantification of the viscosity of the CMT test may allow the 

development of a portable device for more accurate determination of 

SCC cow side and be developed for an in-line device in automated 

milking systems. 

44 Prevalence of Intramammary Infections in Heifers around 

Calving in 40 Dairy Herds in the West of France. 

L. Durel 1 ,C. Guidarini 2 , P. Moroni 3 , C. Locatelli 3 , L. Scaccabarozzi 3 , 

E. Schmitt-Van de Leemput 4 

1 

Clinique Vétérinaire de Marigny, Marigny, France 

2 

Boehringer Ingelheim France, Technical & Development 

Department, Reims, France 

3 

Facolty di Medicina Veterinaria di Milano, Patologia animale, 

Igiene e Sanit’Pubblica Veterinaria, Milano, Italy 

4 

Clinique Vétérinaire de Villaines-la-Juhel, Villaines-la-Juhel, 

France 

Intramammary infections (IMI) in dairy heifers around calving are 

responsible for important economical losses. A better understanding 

of the prevalence, the responsible germs and the risk factors of such 

infections is needed to improve heifers udder health status in France. 

To investigate the prevalence and the risk factors of IMI in heifers, 

an epidemiological study has been carried out in two veterinary 

practices in the west of France from June to December 2007. In this 

abstract, the bacteriological data are reported. Sterile milk samples 

of the 4 individual quarters (n=1.496) were collected of dairy heifers 

(n=378) between 3 and 7 days post partum from 35 dairy herds and 

frozen until analyses. For each sample, somatic cell count was 


determinate and bacteriological analyses were performed according 

to the recommendations of the NMC and the IDF. 1% of the quarters 

(n=16) were not functional at calving time. In total, 22.3% (n=333) 

of the milk samples was positive in bacteriological analyses, 77.3% 

(n=1.156) was negative; 2.1% of the samples was contaminated. In 

general there was no clear relationship between elevated cell count 

(> 100.000 cells/ml) and positive result in bacteriological analyses. 

However, when the data were analyzed per type of bacteria, it 

showed that the presence of Streptococcus spp. (Streptococcus 

uberis) coincided in 77% (85%) of the cases with an elevated cell 

count, whereas the presence of Staphylococcus spp in 48% (S. 

aureus) and 64% (CNS). Among infected heifers (n=214), the 

number of animals with 1, 2, 3 or 4 quarters infected, were 

respectively, 63.0%, 22.4%, 10.3% and 4.2%. 43.4% (n=164) of the 

heifers didn’t have any quarter infected. The frequency of presence 

of CNS, Staphylococcus aureus, Streptococcus spp (Streptococcus 

uberis) and Gram negative spp. were respectively 64.2%, 8.4%, 

16.2% (9.3%) and 4%. On the herd level, the lowest prevalence of 

infected animals was 20% and the highest 100%, but for most of the 

farms (n=26) the infection rate varied from 45 to 75 %. In 

conclusion, IMI in heifers has a high prevalence in France and most 

of the herds have numerous infected animals at calving. Further 

analyses of the data are needed to provide more information on the 

economical impact and the risk factors of IMI infections of heifers. 

45 Bovine Mastitis: Genotypes of Staphylococcus aureus Differ in 

Their Clinical Outcome after Intramammary Infection 

HU. Graber 

Clinic for Ruminants, Vetsuisse Faculty, University of Bern, Berne, 

Switzerland 

Objectives of study: Based on our clinical experience on bovine 

mastitis, we hypothesized that there are subtypes of Staphylococcus 

aureus (S. aureus) which differ in their contagious and pathogenic 

properties after intramammary infection (IMI). 

Materials and Methods: 210 strains of S. aureus were isolated from 

bovine milk samples collected during a representative epidemiological 

study on S. aureus in cow herds of Switzerland. In each of the 26 herds 

investigated, all lactating cows (n=449) were evaluated clinically for 

udder health. Milk of each quarter was then aseptically collected for 

bacteriological testing and analysis of somatic cell counts (SCC). The 

genotypes of S. aureus were determined by PCR amplification of the 

16S-23S rRNA intergenic spacer (RS-PCR). 

Results: In total, 17 genotypes were obtained. Two of them (type B 

and C) were predominant (81%). Each of the remaining types (OG) 

occurred at a very low frequency. Considering genotype B, IMI was 

always present in many cows of a herd. Frequently more than 1 

quarter per cow was infected. In the case of genotypes C and OG, 

IMI was restricted to 1 or very few cows per herd. Constantly, not 

more than 1 quarter per cow was infected. Cow prevalence of S. 

aureus was highly associated with the genotypes (p

2 University of Veterinary Medicine Hanover, Foundation, Institute of 

Food Quality and Food Safety, Hanover, Germany 

Total cell count in milk is used as an indicator of udder health status, 

with each cell type having its own specific function in the immune 

response. The aim of this study was to explore the bovine mammary 

gland defence mechanisms by determining the cellular composition of 

milk in relation to lactation stage, milk fractions and udder health. Total 

cell count (SCC) and differential cell count (DCC) of neutrophils 

(PMNL), lymphocytes (LY), and macrophages (MA) were determined 

on quarter foremilk and stripping samples of 24 dairy cows (08 each in 

early, mid and late lactation). SCC was measured by Fossomatic 

method. DCC were made on cytocentrifuge preparations of cells 

stained with toluidine blue (2 mg/l). Culturing of samples followed the 

procedure of National Mastitis Council. Udder health status was 

defined according to International Dairy Federation. Three grouping 

variables were considered in statistical analysis; milk fraction, stage of 

lactation, and udder health. Results revealed 10%, 5% and 21% of the 

quarters suffering from specific, non-specific and latent mastitis, 

respectively. Udder health and milk fractions showed significant effect 

on cell counts. Even the latent infections caused by Corynebacterium 

and CNS resulted in a significant increase in SCC. Thus, importance of 

these so-called minor pathogens in defining udder defence mechanisms 

may not be ignored. Investigation of interdependence of udder quarters 

indicated significant differences for SCC between healthy and diseased 

quarters of identical udders. Moreover, SCC for healthy quarters from 

healthy udders differed significantly from healthy quarters of udders 

having one or more diseased quarters, thus putting a question mark on 

the independence of quarters. SCC increased towards stripping, and 

this increase was more pronounced in diseased quarters. Proportions of 

PMNL, LY and MA did not change between foremilk and stripping, 

neither in healthy nor in mastitis quarters. Udder health showed 

significant affects on DCC; LY were highest in healthy quarters while 

PMNL in mastitis quarters. Over the lactation, in healthy quarters, 

PMNL and MA proportions increased, and LY decreased. While in 

mastitis quarters, cell proportions remained unchanged. Further, as 

indicated by DCC vis-a-vis SCC, significant pathological/ immunological 

changes seem to occur at 100x10 3 cells/ml. Thus, probably, the 

physiological limit for SCC of healthy udders in bovines may be 

defined even less than 100x10 3 cells/ml. 

77 Evolution of CMSCC in Intramammary Staphylococcus 

aureus Infected Cows after Calving and Treated at Dry Period 

C. Ribeiro 1 , J. Galí 1 , D. Castro 1,3 , P. Teixeira 1 , A. Casas 3 , 

B. Fernández 3 , E. Barrio 3 , J. Simoes 3 

1 Centro Veterinário de Aveiro, Aveiro, Portugal 

2 Grup Tecnic Veterinari S.L., Gerona, Spain 

3 University of Trás-os-Montes e Alto Douro, Veterinary Science 

Department, Vila Real, Portugal 

The aim of this work was to characterise the evolution of somatic cell 

count of mastitic cows infected with Staphylococcus aureus treated only 

at dry period. Composite milk somatic cell counts Somatic cell count 

(CMSCC, fossmatic) and microbiologic analysis (Plate Count Agar) 

were performed, at same time, in all 134 lactating cows from one dairy 

farm. All cows diagnosed with St. aureus infection were managed 

separately and only treated with betalactamic antibiotics at the onset of 

dry period. The CMSCC of first and second dairy report after calving was 

also used. Linear score (LS), bulk contribution (SCC %) of each animal 

and mean linear score (mLS; average of three last individual LS) were 

evaluated for each sample of composite milk. The LS was 3.4±0.2 

(S.E.M., n=73) and 1.9± 0.2 (n=61, P

were 0.17; 0.13 °C (Max.: 0.73). During 24 hours a diurnal variation of 

meanT was visible with a maximum of +0.45; 0.58 °C at 9:30 pm and 

a minimum of -0.42 - 0.39 °C at 6:30 am compared to an individual 

mean temperature calculated for 24 hours. Influence of the ambient 

temperature was statistically significant, but its effect was minimal. 

After infection (pi) both hind quarters showed a significant elevation in 

temperature, which was more pronounced in maxT than in meanT 

between 13 to 17 hours pi compared to values measured 24 hours before 

(maxT at 13 hours pi: infected quarter: +1.98; 0.59 °C; uninfected quarter: 

+1.93; 0.62 °C, p = 0.04 for both). Comparing meanT between 

hindquarters the difference was greatest at 13 hours pi and temperature 

was lower (-0.90; 0.64 °C) at the infected quarter. Differences between 

maxT in hindquarters were not significant at any time point. 

Conclusions: After experimental infection of the udder with E. coli 

significant changes due to inflammation can be measured by 

thermography. 

80 Economic Assessment of Selective versus Blanket Dry Cow 

Treatment Options Including Teat Sealer Use 

H. Seegers 1 , D. Billon 1 , P. Roussel 2 , F. Serieys 3 , N. Bareille 1 

1 

Veterinary School & INRA, Animal Health Management Group, 

Nantes, France 

2 

Institut de l’Elevage, Angers, France 

3 

Filiére Blanche, Rennes, France 

Blanket antibiotic dry cow treatment has become challenged. This is of 

course to relate with residues and resistance, but also with more 

questions on its necessity, especially in herds with a low prevalence of 

intramammary infections at drying-off and a low risk for new 

infections during the dry period. Moreover, the use of an inert teat 

sealer can help to limit antibiotics use. However, no grounded 

indications for selective antibiotic treatment implementation (or teat 

sealer use alone or in combination with antibiotics) are available. 

A dynamic stochastic simulation model was used to assess several 

selection rules (using individual milk SSC thresholds from 100,000 to 

300,000 cells/mL) for cows left untreated, subjected to antibiotics or 

treated with teat sealer. Simulated initial epidemiological conditions in 

a typical 50-cow herd covered a range of variation in type of pathogens, 

prevalence at drying-off, and risk of new infections during the dry 

period. The utility function to rank the rules combined: (i) gross 

margin, (ii) total number of antibiotic treatments per cow-year, and (iii) 

number of clinical cases per cow-year. Simulations assumed that 

blanket antibiotic therapy was replaced by another control option and 

the outcomes with this new control option were compared to those 

obtained with keeping on blanket therapy. 

On short-mid term horizons (3 

years), detrimental evolution was usually observed, justifying 

sometimes a return to blanket therapy. For options including the teat 

sealer use, neutral economic effects were mostly found, but also some 

very positive economic effects for specific high prevalence and high 

risk situations (especially with large implication of environmental 

pathogens). Finally, a decision chart based on prevalence of infections 

at drying-off, risk for new infections during the dry period and type of 

pathogens predominantly involved was proposed. 

Key words: dairy cow, dry-cow treatment, antibiotics, teat sealer 

81 Risk Factors for Bacteriological Quality of Bulk Tank Milk in 

Prince Edward Island Dairy Herds 

A. Elmoslemany, G. Keefe, I. Dohoo 

Atlantic Veteirnary College, Health Management, Charlottetown, 

Canada 

The objective of this study was to determine on-farm risk factors for 

bacteriological quality of bulk tank milk. Bulk tank raw milk quality was 

evaluated on all Prince Edward Island dairy herds (n=235) over a two 

year period (March 2005-March 2007). Biweekly total bacterial, 

preliminary incubation, laboratory pasteurization and coliform counts, 

were conducted using Petrifilm ® . For the assessment of risk factors, a 


case control study was conducted from January 2006 to May 2007. Case 

and control herds were defined based on the last 6 results of bulk tank 

bacterial count prior to on farm evaluation. A total of 69 herds (39 cases 

and 30 controls) were evaluated. Data collection included a survey of 

management practices, cow, environment and equipment hygiene 

scoring, milking procedures and mastitis control, complete wash analysis 

of the milking equipment and monitoring the presence of bacterial films 

on equipment. Data was analyzed using multivariable logistic regression. 

A total of 16 variables were associated with bulk tank milk bacterial 

quality at p

2 Schering Plough Animal Health, R&D, Wellington, New Zealand 

Streptococcus uberis is common cause of clinical and subclinical 

mastitis. The objective of this study was to assess the efficacy of 

vaccination of dairy cows with a whole-cell killed vaccine containing 

S. uberis on the incidence and prevalence of mastitis associated with S. 

uberis. Cows from 3 dairy herds were either vaccinated with a vaccine 

containing S. uberis (n=695) or injected with the vaccine vehicle 

(n=711 cows) on 2 occasions approximately 4 wk apart with the 2 nd 

vaccination occurring on average 55 days before calving. Milk samples 

were collected from all 4 glands from a randomly selected 2/3rds of the 

enrolled cows between 0 and 7 days after calving, and milk samples 

were collected from cows with clinical mastitis (CM) within 21 days of 

calving. The prevalence of intramammary infection associated with S. 

uberis and the incidence of CM associated with S. uberis were analysed 

at cow and gland level using multivariable models which accounted for 

the clustering of gland with cow and cow with herd and adjusted for the 

effect of potential confounders such as age and breed. Vaccination 

tended to reduce the prevalence of cows infected with S. uberis in any 

gland within seven days of calving (0.12 (0.09-0.16) vs. 0.16 (0.13- 

0.21) for vaccinated vs. control cows, respectively; p=0.057). 

However, there was no difference in the prevalence of S. uberis at 

gland level (0.043 (0.031-0.059) vs. 0.055 (0.042-0.072) for vaccinated 

vs. control cows, respectively; p=0.18; RR=0.785 (0.548-1.124)) or in 

incidence of CM associated with S. uberis between the groups at either 

cow (43/670 (6.4%) vs. 44/693 (6.3%) in vaccinated vs. unvaccinated 

cows, respectively; p=0.95) or gland (53/2628 (2.0%) vs. 48/2724 

(1.7%) in vaccinated vs. control cows, respectively; p=0.50) level. 

Vaccination reduced the number of S. uberis present in milk in glands 

from which it was isolated (940 (532-1661) vs. 2476 (1200...) 

geometric mean colony forming units/ml for vaccinated vs. control 

cows, respectively; p=0.013). There was no difference in SCC at first 

herd test between vaccinated and control cows (71 (64-78) vs. 75 (68- 

83) geometric estimated marginal mean (95% CI) x 103 SCC/ml for 

vaccinated and control cows, respectively; p=0.32). It was concluded 

that the whole-cell killed S. uberis vaccine evaluated in this study was 

not effective in reducing the prevalence and incidence of mastitis 

associated with S. uberis. 

84 Prototheca blaschkeae Associated with Bovine Mastitis 

S. Marques 1 , E. Silva 1 , C. Kraft 2 , V. Huss 2 , G. Thompson 1 

1 Universidade do Porto - ICBAS, Department of Veterinary Clinics - 

Laboratory of Infectious Diseases, Vila do Conde, Portugal 

2 Friedrich-Alexander - Universität Erlangen-Nürnberg, Department 

für Biologie, Molekulare Pflanzenphysiologie, Erlangen, Germany 

Objectives of study: Algae of the genus Prototheca and more rarely 

Chlorella are the only known plants that cause infectious diseases in 

humans and animals. Prototheca are ubiquitous, generally saprophytes, 

and can be isolated from a great variety of environmental sources. Until 

recently only two species were known to be associated with disease in 

animals and humans, Prototheca zopfii and P. wickerhamii, 

respectively. In 2006, a new species P. blaschkeae was described, 

which has been isolated from a case of disease in men. The prevalent 

form of protothecosis in animals is bovine mastitis, which generally 

occurs in a chronic subclinical or a mild clinical inflammatory process 

of the udder affecting cows that do not respond to routine therapy. The 

objective of the present work was to characterize Prototheca spp. 

isolated from bovine mastitis. We provide evidence for the first time, 

that P. blaschkeae is associated with bovine mastitis. 

Material and methods: Forty one Prototheca isolates obtained from 

mastitic milk from dairy cows belonging to 22 different farms in the 

North of Portugal were characterized by conventional microbiological 

methods and by phylogenetic analysis of their 18S rDNA sequences. 

The methods employed were morphological data analysis, 

biochemistry profile determination by API 20C Aux and BBL Crystal 

Kits (Enteric Nonfermenter and Gram positive) systems, PCR 

amplification of the 18S rDNA, amplified rDNA restriction analysis 

with HaeIII, and phylogenetic analysis of the 18S rDNA sequences. 

Results: Macroscopical and essentially microscopical analyses 

revealed two groups of algae, one that contained the majority of the 

isolates and the other with only four isolates. Both groups were 

confirmed by their biochemical profiles. PCR amplification of 18S 

rDNA and phylogenetic analyses showed, that most of the isolates were 

identified as P. zopfii var. hydrocarbonea, whereas the four isolates of 

the other group were identified as P. blaschkeae. 

Conclusion: Although the type of mastitis induced by all Prototheca 

isolates had similar clinical manifestations, we show for the first time, that 

besides P. zopfii also P. blaschkeae may be involved in bovine mastitis. 

Key words: bovine mastitis, Prototheca, genotyping 

127 Controlling Highly Prevalent Staphylococcus aureus Mastitis 

from the Dairy Farm 

H. Nagahata 1 , H. Maruta 1 , S. Matsuki 1 , H. Higuchi 1 , K. Kawai 2 , 

A. Anri 3 

1 

Rakuno Gakuen University, Department of Animal Health, Ebetsu, 

Japan 

2 

Tokachi NOSAI, Seibu veterinary clinical center, Obihiro, Hokkaido, 

Japan 

3 

Hokkaido NOSAI, Unit of Animal Health Management, Ebetsu, 

Japan 

Objective: Staphylococcus aureus, a contagious pathogen that spreads 

easily from cow to cow, usually manifests as subclinical mastitis. A 

dairy farm with increasingly elevated somatic cell counts (SCC) in the 

bulk milk and a high rate of S. aureus-mastitis was referred. Our study 

monitored the herd for 3.5 years after we instituted a mastitis control 

program. In addition, to evaluate whether a high prevalence of S. 

aureus infection in lactating cows affects the occurrence of S. aureus 

infection in heifers in the herd, bacteriological findings in mammary 

secretions from heifers were examined. 

Methods: Dairy farm had 57 lactating Holstein cows were housed in freestall 

barns. A total of 15 heifers, 23-26months old, were also used for the 

survey of microbiological findings in mammary secretions. Quarter milk 

samples from lactating cows were cultured, and pathogens were identified. 

Mastitis control program: To determine the contributory causes of S. 

aureus infection, we checked the milking procedures and implemented the 

following mastitis control measures: (a) maintenance of the correct 

milking order, (b) recommended milking procedures, (c) post-milking teat 

dipping with approved disinfectants, and (d) culling of cows infected with 

S. aureus in 2 or more quarters or cows having chronic S. aureus infection. 

Results: S. aureus was detected in 24 (41.2%) of the 54 lactating cows 

and in 29 (12.8%) of 227 quarters of the 57 milking cows in the herd. The 

S. aureus infection rate decreased steadily from 42.1% at the time of our 

first visit to 10.5% seven months after start of the control program, then 

decreased to 4% at 14 months, and no isolate of S. aureus was found in 

milk samples at 18 months. In the course of the 3.5-year monitoring 

period, 24 lactating cows infected with S. aureus were culled. The average 

SCC in the bulk tank milk decreased gradually from 52.3 x 10 4 cells/ml 

(21 to 94 x 10 4 cells/ml ) to 14.5 x 10 4 cells/ml (4 to 21 x 10 4 cells/ml), as 

observed 18 months after start of the control program. No S. aureus isolate 

was detected in 58 mammary secretions from 15 heifers at 4-5 weeks 

before parturition. 

Conclusion: The program implemented is effective in controlling S. 

aureus mastitis on the dairy herd. A high S. aureus prevalence in the herd 

may not necessarily be a decisive factor for S. aureus infection in heifers. 

Key words: long-term monitoring, mastitis control program, S. aureus 

128 The Recoveries from Acute Mastitis and Fertility Indices of 

Cows Treated with Antibiotics and Additional Medicine 

E. Malinowski, S. Smulski, H. Markiewicz 

National Veterinary Research Institute, Department of 

Pathophysiology of Reproduction and Mammary Gland, Bydgoszcz, 

Poland 

The aim of studies was to establish the effect of antioxidants, 

immunomodulator or NSAID single injection on results of mastitis 

therapy and fertility indices of mastitic cows treated intramammarily with 

antibiotics. Examinations were performed in 4 farms on 240 dairy cows 

that showed acute form of mastitis as only clinical disease between 10 th 

and 100 th day after parturition and on 60 healthy cows calved at the same 

time. Mastitic cows were treated with approved intramammary products 

in doses recommended by manufacturers. Animals included to I group 

received only intramammary therapy (IMT). Cows from group II, III and 

IV, apart from IMT, were once injected with antioxidants (vit. C, vit. E, bcarotene, 

Se), lysozyme dimer) or flunixin meglumine, respectively. 

Healthy cows from group V consisted the control for fertility indicators of 

mastitic animals. In average, 55.6% of cows recovered as an effect of the 

first course of the intramammary treatment. A single injection of 

antioxidants or lysozyme dimer increased recovery rates. Fertility indices: 

calving to the first service, conception rate, insemination index and 

Mastitis / Udder Health and Milk Quality 49

calving pregnancy period were the best in the control group. The single 

injection of the additional medicine increased fertility indices if compared 

to cows treated intramammary only. Fertility indices were better in cows 

recovered as the effect of the first treatment and worse in cows that 

became chronically inflamed and required additional therapy of mastitis. 

It can be concluded: acute mastitis between 10th and 100th day 

postpartum elongates the period from calving to first insemination, 

decreases the conception rate, and increases the insemination index, 

calving pregnancy period and culling rate of dairy cows. More detrimental 

are the chronic cases than acute, successfully treated cases. The single i.m. 

injection of antioxidants (vit. C, vit. E, b-carotene, Se) or the lysozyme 

dimer increases the efficacy of intramammary antibiotic treatment of 

mastitis caused by Streptococcus spp., CNS, E. coli and even Staph. 

aureus. It seems the i.m. injection of flunixin meglumine increases the 

efficacy of coliform-mastitis therapy with antibiotics. However injection 

of additional medicine to intramammarily treated cows sick with acute 

mastitis improves their fertility indicators. 

Key words: cows, mastitis, antibiotics, antioxidants, lysozyme dimer, 

NSAID, fertility 

129 The Contagious Staphylococcus aureus Genotype B Affects 

the Presence of Other Mastitis-Relevant Bacteria in Dairy 

Herds 

A. Michel, A. Steiner, HU. Graber 

Clinic for Ruminants, Vetsuisse Faculty, University of Bern, Berne, 


Objectives of study: Staphylococcus aureus (S. aureus) is globally 

one of the most important pathogen of subclinical mastitis and causes 

large economic losses. In the present study, the effect of the contagious 

S. aureus genotype B on the presence of other mastitis-associated 

pathogens (coagulase-negative staphylococci (CNS), Streptococcus 

spp. (non-agalactiae streptococci), and Corynebacterium spp.) was 

investigated in Swiss dairy herds. 

Materials and methods: Sample herds (SH): 1057 milk samples from 

265 cows (10 farms) with diagnoses of S. aureus genotype B problems 

were collected. As control herds (CH), 257 cows (20 farms with 1021 

milk samples) were selected with at least 1 S. aureus isolate of non-B 

genotype (genotypes A, C, H, E, D, R). All the samples were 

bacteriologically analyzed according to standard guidelines (NMC). 

Genotyping was done by ribosomal spacer-PCR. For the SH and CH, 

the prevalences on cow and quarter levels of S. aureus, Streptococcus 

spp., CNS and Corynebacterium spp. were compared using nonparametric 

statistics. Binary logistic regression was used to select those 

variables which best classified SH and CH. 

Results: Considering S. aureus, the cow and quarter prevalences 

clearly differed between SH and CH (p

167 Genotyping of Staphylococcus aureus Isolated from Bovine 

Mastitis Cases and Asymptomatic Carrier Mammary Glands 

F. Bispo 1 , I. Rivera 1 , J. Arcaro 2 , E. Mamizuka 1 , C. Mendonça 3 , 

P. Melville 1 , E. Costa 1 

1 University of Sao Paulo - Institute of Biomedical Sciences, 

Microbiology, sao Paulo, Brazil 

2 Instituto de Zootecnia, Bovinocultura Leiteira, Nova Odessa, Brazil 

3 Universidade Federal Rural de Pernambuco, Departamento de 

Medicina Veterinária Preventiva, Recife, Brazil 

The mastitis caused by Staphylococcus aureus is one of the main diseases 

in dairy herds and is responsible for chemical, physical, and 

microbiological alterations in milk, resulting in serious economical losses 

and public health risk. Chronically infected cows are the main reservoir of 

infection in a herd; however, there are other contamination sources and 

reservoirs, such as milking unit liners, and human and bovine skin. 

Additionally, the carrier status is characterized by mammary glands 

without clinical signs of inflammation, negative CMT reaction, and 

SCC

2 University of Nottingham, Department of Veterinary Medicine, 

Nottingham, United Kingdom 

3 Orchard Veterinary Group, Glastonbury, United Kingdom 

4 Quality Milk Management Services Ltd., Wells, United Kingdom 

Objectives: The primary objective of the study was to determine the 

impact of the duration of treatment on the efficacy of penethamate 

hydriodide (Mamyzin ® Injection) in the cure of persistent bovine intramammary 

infection in U.K. dairy cows. 

Materials and Methods: Cows with a history of a high somatic cell count 

(SCC), four functional quarters, no significant teat lesions and in good 

general health were enrolled into a treatment study (n=95). Two different 

regimes were assessed using penethamate hydriodide, which was 

administered either as a four-day (“on-label”) course or a seven-day 

(“extended”) course. Intra-mammary infection status was monitored 

using both bacteriology and quarter somatic cell counting from 1-week 

pre-treatment until 4-weeks post-treatment. Multi-level statistical models 

were specified and interpreted within a Bayesian framework to estimate 

the probability of a difference between the two treatment groups 

(expressed as an odds ratio), using a flat, normal prior distribution. 

Results: Preliminary results have shown that both treatment groups 

experienced a significant reduction in the prevalence of gram-positive 

major pathogens from before until after treatment accompanied by a 

fall in somatic cell counts at both the quarter and cow level. There was 

a reduced odds of identifying a major pathogen after treatment in 

quarters treated for 7 days compared to quarters treated for 4 days 

(mean odds ratio = 0.57) and a posterior probability of 93% that 

treatment for 7 days resulted in fewer quarters with major pathogens 

than treatment for 4 days. There was an increased odds that quarters 

would remain below 200,000 cells/ml for 28 days after treatment for 7 

days compared to treatment for 4 days (odds ratio = 1.87) and a 

posterior probability of 79% that treatment for 7days resulted in fewer 

quarters > 200,000 cells/ml than treatment for 4 days. 

Conclusion: This study demonstrates the efficacy of Mamyzin ® 

Injection in the treatment of persistent IMI in dairy cows in the U.K. 

and has identified an increased probability of a cure when using a 7day, 

extended therapy regime, compared to 4-day, on-label treatment 

regime. In addition, the results can be used to inform clinical decisionmaking 

for the treatment of high SCC cows. 

Key words: somatic cell count, penethamate hydriodide, extended 

therapy, persistent infection, streptococcus uberis 

172 Challenge with Streptococcus uberis to Evaluate the Efficacy 

of Teat Sealers in Preventing Mastitis 

N. Williamson, K. Petrovski, C. Fernandez 

Massey University, Institute of Veterinary, Animal and Biomedical 

Sciences, Palmerston North, New Zealand 

External teat sealers based on tetrahydrofuran (DryFlex (DF)) and a 

wound seal for humans 2-octylcyanoacrylate (Band-Aid (BA)) to prevent 

dry period mastitis were investigated. Streptococcus uberis has the 

highest incidence in the dry period and early lactation in New Zealand. It 

is a ubiquitous environmental pathogen, is normally easy to cure using 

antibiotics and thus was an appropriate challenge organism. Quarter milk 

samples were taken for bacteriological culture from 175 cows, 4 and 1 

days before dry-off and twice within 4 days after calving. After the last 

milking, 88 cows received DF and 87 cows received BA. Pairs of contralateral 

quarters were randomly assigned to the treated or the control group. 

All quarters were dipped in a Strep. uberis broth 2 and 4 days after dry-off 

and were assessed daily for clinical mastitis. Clinical mastitis iwas 

greatest 6-11 days after dry-off in both groups. Strep. uberis was the most 

prevalent bacterium isolated. In the DF group, 35 cases of clinical mastitis 

occurred in 176 treated quarters, compared with 83 in the 176 control 

quarters. In the BA group, 67 cases of clinical mastitis occurred in 174 

treated quarters, compared with 64 in 174 control quarters. DryFlex reduced 

mastitis after dry-off but not after calving. Octylcyanoacrylate did not 

reduce clinical mastitis after dry-off. A DryFlex type teat sealer showed 

potential in preventing mastitis in the early dry period, but has now been 

withdrawn worldwide. The effect of experimentally induced clinical 

mastitis in the early dry period on the subsequent lactation was examined. 

Cows (127) developing clinical mastitis after challenge were treated with 

antibiotic. Yields of milk, fat, protein and milk solids were analysed with 

respect to treatment group. No significant effect of treatment group on 

lactation yields of milk, fat, protein or milk solids occurred. No difference 

in production occurred (milk yield, 5126 vs. 5010 litres; fat yield, 267 vs. 

264 kg; protein yield, 182 vs. 179 kg) between cows that had mastitis and 


those that did not. Promptly treated clinical mastitis due to Strep. uberis 

did not affect production in the subsequent lactation. The ability to 

demonstrate differences in efficacy and absence of production effects in 

the subsequent lactation mean this challenge model appears suitable for 

evaluating teat sealers. 

Key words: mastits, Streptococcus uberis, teat sealer 

173 Use of Leukocyte Differential Patterns to Optimize Diagnosis 

of Bovine Intramammary Infections: Evaluation of 

Partitioned vs. Non-partitioned Testing 

A. Rivas 1 , K. Anderson 1 , G. Leitner 2 , M. Chaffer 2 , O. Krifucks 2 , 

R. Rodriguez 1 

1 North Carolina State University, College of Veterinary Medicine, 

Raleigh, NC, United States 

2 Kimron Veterinary Institute, Bacteriology,Beit-Dagan, Israel 

Objectives: To explore the use of leukocyte patterns in diagnosing 

infectious mastitis. 

Materials and methods: Quarter milk samples were investigated in 3 

cross-sectional studies conducted in the US (study I, n=120; and study 

III, n=500) and Israel (study II, n=484). The somatic cell count (SCC), 

milk microbiological cultures, and the percentages, total counts and 

relative proportions of lymphocytes, macrophages and neutrophils 

were established. Non-periparturient (studies I and III) and 

periparturient (study II) cows were assessed. Samples were collected 

without (studies I and III) and with previous determination of health 

status (study II). Disease prevalence was >40% (study I) and

total of 31 dairy cows were treated and 30 cows were used as negative 

controls. The overall cow cure rate was 25.8% (8/31) in the treated group 

and 3.3% (1/30) in the control group. The difference between the 2 groups 

was statistically significant (P = 0.013). A tendency to a higher cow cure 

rate was also observed in the treated group when a cow had only one 

quarter infected (36.8%) compared to if more than 1 quarter was infected 

(8.3%) (P = 0.1). The quarter cure rates at 10 d post-treatment were 77.5% 

(38/49) and 17.6% (9/51) in the treated and control groups, respectively (P 

< 0.0001). The overall quarter cure rates were 39.4% (13/33) and 4.5% 

(2/44) in the treated and the control groups, respectively (P < 0.01). 

Antibiotic residues were tested on 13 treated cows. Only 1 cow had 

cephapirin residues above the accepted limit 4 d (96h) after the end of the 

treatment. This cow was under the accepted MRL limit 4.5 d (108h) after 

the end of the treatment period. Based on these results, the suggested milk 

withdrawal time for this study was set at 5 d (120h). 

Conclusion: The cow and quarter cure rates were significantly higher 

in the treatment group than in the control group. The treatment tends to 

be more effective when only 1 quarter was infected. A milk withdrawal 

time of 5 d (120h) was sufficient in all tested cows. 

Acknowledgements: The study was supported by Fort Dodge Animal 

Health. 


549 Study on Frequency, Etiology and Somatic Cell Counts of 

Subclinical Ovine Mastitis in Tabriz, Iran 

R. Beheshti Govij 1 , J. Shaigh 1 , J. Dulgari 1 , M. Mousapour 2 

1 

Islamic Azad University-Shabestar Branch, Veterinary, Shabestar, 

Iran 

2 

East Azarbaijan Burau, Department of Veterinary, Tabriz 

Veterinary Network, Tabriz, Iran 

To investigate the period prevalence, etiology and some 

epidemiological features of subclinical mastitis in ewes from the Tabriz 

region, milk samples from 193 lactating ewes were aseptically 

collected for bacterial and somatic cell counts (SCC) tests. Milk 

samples were collected from April to May of 2007. An association was 

observed between the occurrence of subclinical mastitis and the age of 

ewe. The periodic prevalence rate of SCM was 13.5%. Staphylococci 

were the most prevalent bacteria, representing 88.4% of the isolates. 

Coagulase-negative staphylococci (69.2%), was the most prevalent 

species followed by staphylococcus aureus (19.2%). E.coli and C.bovis 

was the second bacterial group in importance according with the 

distribution among flocks representing 7.7% and 3.8% of the isolates. 

Subclinical mastitis seems to be, as deduced from the high prevalence 

observed in this study, an important health problem for milking sheeps 

in the Tabriz region. 

Key words: subclinical mastitis, sheep, etiology, prevalence 

550 Risk Factors for Subclinical Mastitis in Dairy Cows 

G. Souza 1 , C. Coentrao 2 , J. Brito 1 , M. Brito 3 , W. Lilenbaum 4 

1 National Center for Resarch on Dairy Cattle, Milk Quality 

Laboratoy, Juiz de Fora - MG - Brasil, Brazil 

2 Universidade Federal Fluminense, Faculdade de Veterinária, 

Niterói - RJ - Brasil, Brazil 

3 National Center for Research on Dairy Cattle, Milk Microbiology 

Laboratory, Juiz de Fora - MG - Brasil, Brazil 

4 Universidade Federal Fluminense, Faculdade de Veterinária, Juiz 

de Fora - MG - Brasil, Brazil 

This study was conducted to identify risk factors for subclinical mastitis 

(SCC > 200,000 cells/ml). A total of 2,657 lactating cows from 24 herds 

in the State of Minas Gerais, Brazil, were included in the study. Each of 

the farms was visited three times in an 8-month period from November 

2005 to June 2006. At each visit, all milking cows were examined for 

clinical mastitis by a single observer. A total of 3,987 milk samples were 

examined for somatic cell counts (SCC). Statistic analyses were 

conducted using descriptives statistics and logistic regression models to 

estimate the relative risk (odds ratio = OR). Considering all milk samples, 

the mean, median and standard deviation values for SCC were 

respectively 608,000, 219,000 and 967,000 cells/ml. Risk factors for 

subclinical mastitis were: udder positioned at the same height or below the 

hock (OR=1,73), presence of cracks or fissures in the rubber parts of the 

milking machine (OR=2,45), inadequacy of teat cups (OR=1,48), 

infrequent and ill-defined scheme for cleaning the pulsators (1,41), 

milkers not trained for operating the milking equipments (2,51), no 

information about the mastitis pathogens present in the herd (1,84), 

immersion of teat cups in disinfectant solution between milkings 

(OR=2,19) and total insertion of cannula in teats during antibiotic 

treatment (OR=2,64). The high variation of SCC values (608.000- 

967.000 cells/ml) suggests that others factor such number of infected 

mammary quarters and pathogens enrolled can influenced the results. The 

methodology used did not allow to identify all risk factors that increasing 

SCC. Therefore, the results can also to be used to improve the mastitis 

control programs currently used by these herds. 

Study supported by Fundaçao de Amparoa Pesquisa do Estado de 

Minas Gerais (FAPEMIG). Grant number EDT 401/05 

Key words: bovine, mastitis, risk factors, somatic cell count 

551 Some Epidemiological Studies on Subclinical Mastitis in 

Dairy Cows in Assiut Governorate 

A. Abdel-Rady 1 , M. Sayed 2 

1 

Faculty of Veterinary Medicine, Assiut University, Animal Medicine, 

Assiut, Egypt 

2 

Faculty of Veterinary Medicine, Assiut University, Department of 

Food Hygiene, Assiut, Egypt 

In this investigation, some epidemiological studies were run on 

subclinical mastitis for totally 350 dairy cows of different breeds, ages and 

distributed in different villages in Assiut governorate, Assiut, Egypt, 

along a whole year (during the period from June 2006 till July 2007) 

through field screening surveys by using of the California mastitis test 

(CMT) for each quarter milk sample followed by bacteriological 

examination.The dairy cows were differed from the breed point of view as 

230 Holstein Friesian breed and 120 native breed. Also, they were 

differed from the age point of view as a group of 95 cows aged from 2 to 

4 years old and another group of 255 cow aged from 5 to 8 years old. The 

obtained results revealed that 67 cows (19.14%) had 80 infected quarters 

(5.71%). Prevalence 52.5, 31.25 and 16.25%, respectively. With studying 

the breed factor, it was found Friesian breed was sensitive towards 

infection (20.43% at the cow level and 6.09% at the quarter level) than of 

native breed (16.67% at the cow level and 5% at the quarter level). It was 

also noticed that the prevalence of subclinical mastitis in hot weather as 

during summer (9.14% at the cow level and 2.64% at the quarter level) 

and during spring (4.86% at the cow level and 1.36% at the quarter level) 

was higher than in cold weather as during winter (2% at the cow level and 

0.64% at the quarter level) and during autumn (3.14% at the cow level and 

1.07% at the quarter level). In relation to age susceptibility, 5-8 years old 

cows (15.43% at the cow level and 4.36% at the quarter level) were 

susceptible than those of 2-4 years (3.71% at the cow level and 1.36% at 

the quarter level). The degree of quarter attack according to positive CMT 

was varied from 35 quarters (2.50%) showed degree (+++), to 45 ones 

(3.22%) showed degree (++), to 120 ones (8.57%) showed degree (+) and 

the rest (85.71%) showed degree (-). The obtained results threw the light 

on the epidemiology of subclinical mastitis in Assiut villages and 

provided an importance of the CMT for diagnosis of subclinical mastitis 

due to it is a reliable, easy, rapid and cheap tool helping in diagnosis and 

controlling the disease. Programs for control of subclinical mastitis may 

be planned around the routine examination of all lactating cows, and 

consequently early treatment can be applied towards positive cases 

rapidly for preventing their conversion towards clinical form among dairy 

cows and for protecting the herd health, milk hygiene and consequently 

the consumer health. 

Key words: epidemiology, subclinical mastitis, California Mastitis 

Test (CMT), intramammary infection (IMI), dairy cows 

552 Field Trial in Nothern Italy to Evaluate the Efficacy of Two 

Different Therapies against Acute Clinical Mastitis in Dairy 

Cows 

V. Cuteri 1 , R. Froyman 2 , L. Nisoli 3 , A. Romero Tejeda 1 , 

S. Preziuso 1 , A. Attili 1 

1 University of Camerino, Veterinary Science, Matelica (MC), Italy 

2 Bayer Health Care, Animal Health Division, Monheim, Germany 

3 Bayer Health Care, Animal Health Division, Milan, Italy 

This study compared the efficacy of two different treatment regimens 

against acute clinical mastitis cows by enrolling 155 clinically ill dairy 

cows into a multicentric, controlled and randomised trial. One group (76 


animals) was treated with 5 mg enrofloxacin/kg BW (Baytril ® 10% inj. 

solution) intravenously for 3 consecutive days while the other group (79 

animals) was treated with 1 intramammary syringe of Cobactan ® LC (75 

mg cefquinome)/mastitic quarter three times every 12 hours and 

concurrently with one daily intramuscular injection of 1 vial Mamyzin ® 

(equivalent to 10 grams of penethamate/head) for 3 consecutive days. The 

animals were followed for 14 days and received a daily physical 

examination of the udder on days 1 (enrolment day and first day of 

treatment), 2, 3, 5, 7 and 14. The rectal temperature, the udder secretion, 

and general symptoms were evaluated. Sterile samples of milk were 

collected on days 1, 5, 7 and 14 and were cultured to reveal the presence 

of pathogens. Further, the effect of treatment on the incidence of clinical 

mastitis, on bacteriological cure and on somatic cell counts was examined. 

The most frequently isolated microorganisms were Str. uberis (31/155, 

20%) and E. coli (20/155, 13.0%). Both therapies resulted in a significant 

reduction (p 1.200.000 cells/ml with 29 (4.92%) dairies for the first year 

and 11 (1.71%), for the second year. 

555 A Study to Measure the Effect of Using a Sanitizer to Clean 

Cows’ Teats before Milking, on the Bacterial Count of Teat 

Skin 

K. Fehlings 1 , R. Huber-Schlenstedt 1 , J. Deneke 1 , A. Gangl 2 

1 

Bavarian Animal Health Service, Udder Health Service, Poing, 

Germany 

2 

Bavarian Animal Health Service, Bacteriology, Poing, Germany 


The impeccable hygienic quality of milk basically is defined by the 

content in saprophytic, human pathogenic and animal pathogenic bacteria. 

To ensure this bacteria have to be as low as possible. In three Bavarian 

dairy farms the sanitizing and cleaning of teats (60 randomly chosen 

cows) before milking with a 0.5% (750 ppm active chlorine) 

Tosylchloramide sodium solution (Desinficin ® CL; DeLaval GmbH, 

Germany) applied to one-way paper tissues was carried out. A significant 

reduction of the total bacterial count (min. 98.8%, max. 99.3%, average 

99.0%) on the teat skin as well as an effective reduction of the share of 

specific mastitis pathogens was shown. For use the residues of the 

disinfectant solution were squeezed out and the wet udder paper tissue 

used. Two control groups cleaned with one-way paper tissues showed a 

reduction of 87.5% (min. 82.5%, max. 92.7%). For demonstration of the 

bacteria reducing effect the average values of the logarithms were formed 

and the relative bacteria reduction by the treatment was calculated. The 

bacteria per 2 cm2 before treatment were between 7.3 x 10 3 cfu (log 3.87) 

und 5.2 x 10 5 cfu (log 5.72) and after treatment between 1.4 x 10 3 (log 

3.15) cfu and 7.5 x 10 2 cfu (log 2.88). In the control group the bacteria 

were between 3.0 x 10 4 cfu (log 4.48) und 8.7 x 10 5 cfu (log 5.94) and 

between 1.6 x 10 3 (log 3.20) cfu and 9.3 x 10 4 cfu (log 4.97). A teat 

hygiene score (score 1 no growth/< 0.5 cfu, score 2 < = 10 - 30 cfu, score 

3 < = 30 - 50 cfu, score 4 < 50 cfu) was used to show the reduction of 

contaminated teats (CNS, streptococci). The share of teats contaminated 

with mastitis pathogens decreased from score 2-4 corresponding to score 

1 (70.4% resp. 72.5% remaining bacteria). The aim of this study was to 

check the positive effect of Tosylchloramide sodium solution on the 

reduction of bacteria on the teat skin under practical conditions 

(“sanitizing cleaning”) in order to evaluate this approach with respect to 

the reduction of an infection risk of the mammary gland with special 

consideration of mastitis pathogens. Desinficin ® CL is registered for 

sanitizing and cleaning of teats before milking and as drug for teat 

disinfection after milking. Probably it is a method eliminating dirt and 

faeces on the surface of the teats, but of appropriately reducing undesired 

bacteria from the udder/teat skin as far as possible. 

Key words: milk quality, udder cleaning, sanitizing cleaning, 

Tosylchloramide sodium solution, teat disinfection 

556 Aflatoxin Contaminations in Milk and Feed in Kermanshah, 

Iran 

N. Jamali 1 , M. Moeini 1 , M. Ghasempour 2 

1 Razi University, Animal Science, Kermanshah, Iran 

2 Faculty of Veterinary, Animal Medicin, Kermanshah, Iran 

Alfalfa and corn silage and milk samples of 9 industrial and semiindustrials 

of dairy farms in Kermanshah were collected during 3 

months experiment. Feed samples extracted with methanol: water 

(70:30 V/V) and analyzed by Enzyme linked Immune Sorbent Assay 

(Elisa) method. Milk samples collected from dairy herd and aflatoxin 

M1 (AFm1) measured by Elisa method. Blood samples were collected 

by tail venipuncture from %10 cows in each farm for measuring 

biochemical and hematological parameters.The result showed that the 

contamination AF in feed samples in small, medium and big dairy 

farms determined were: 12.47, 10.32 and 11.24 µg/kg respectively. The 

average of AF in January and end of February were: 9.392, 13.29ng/Kg 

DM. There was a significant different between aflatoxin in small and 

big size farm (p

and 5.86 % on quarter basis respectively. The hind quarters were more 

frequently affected than the fore quarters 

Bacteriological examination of milk samples revealed that staphylococci 

were the chief etiological agents both in clinical and sub clinical mastitis 

(41.67%) in camels, followed by Streptococcus spp. (21.67%), 

Enterobacter spp. (15.00%), C. pyogenes (10.00%), Micrococcus spp. 

(5.00%) , Pasteurells spp. (5.00%) and Pseudomonas aeruginosa (1.66%). 

Most of the Staphylococcus spp., Streptococcus spp. and C. pyogenes 

strains were sensitive to carbenicillin, gentamycin , kanamycin, and 

erythromycin, but resistant to colistin and sulphamethoxazole. The other 

mastitis pathogens like Enterobacter, Micrococcus, Pasteurella spp. and 

Ps. aeuroginosa isolates were showed variable pattern of sensitivity to the 

antimicrobial agents. 

558 Comparison of Pathogens and Antimicrobial Susceptibility of 

Isolates from Conventional and Robotic Milking Herds in 

Korea 

D. Kim, S.Y. Jeoung, S.J. Ahn 

Kangwon National University, School of Veterinary Medicine, 

Chuncheon, Kangwon, South Korea, 

In Korea, robotic milking system is introduced about 2 years before. 

However, mastitis control program on the robot milking dairy herds are 

not fully understood. An observational study was conducted to compare 

the pattern of pathogens and the antimicrobial susceptibility of 

Staphylococcus spp. from 8 conventional (pipeline of parlor milking) and 

8 robotic milking dairy herds. All herds composed with Holstein cows and 

the robotic milking herds are using robotic milking system for at least 1 

year before the start of this study. For each robotic milking farm, a 

neighboring conventional dairy farm served as a control. After bulk tank 

milk samples were collected, samples cultured on blood-esculin agar and 

Baird-Parker agar. The cultured staphylococci colonies were identified as 

Staphylococcus aureus and coagulase negative Staphylococcus (CNS) 

by tube coagulase test. Regardless of the milking system, Staphylococcus 

aureus was mainly cultured from the dairy farms in which teat dipping is 

not completely conducted and Streptococcus uberis was cultured from the 

farms in which udder washing was not completely conducted. The total of 

119 Staphylococcus spp isolates was tested for minimum inhibition 

concentration (MIC) against 14 antimicrobial agents by microdilution 

method. The concentration that inhibits 90% (MIC90) of the analysed 

strains in micrograms per mililiter, for S. aureus were 0.5, 256, 16, 4, 1, 

0.25, 1, 32, 8, 0.5, 64, 512, 1 and 0.25 to ampicillin, bacitracin, 

cephalothin, chloramphenicol, norfloxacin, erythromycin, gentamicin, 

kanamycin, oxacillin, penicillin, streptomycin, sulphamethoxazole, 

tetracycline, and vancomycin, respectively. And the MIC 90 values for 

CNS were 1, 64, 0.5, 4, 1, 0.25, 0.5, 2, 0.5, 0.5, 4, 1024, 16 and 0.5 to 

ampicillin, bacitracin, cephalothin, chloramphenicol, norfloxacin, 

erythromycin, gentamicin, kanamycin, oxacillin, penicillin, streptomycin, 

sulphamethoxazole, tetracycline, and vancomycin, respectively. Resistant 

S. aureus isolates were found only to to ampicillin, bacitracin, oxacillin, 

streptomycin, sulphamethoxazole, and penicillin on the basis of NCCLS 

interpretive standards. Significant differences in antimicrobial 

susceptibility between robotic and conventional milking dairies were 

founded only to tetracycline. 

Key words: robotic milking, Staphylococcus spp, antimicrobial 

susceptibility, microdilution method, Korea 

559 Use of Quarter Milk or Composite Samples for Diagnosis of 

Subclinical Bovine Mastitis in Dairy Farms 

C. Ribeiro 1 , D. Castro 1,3 , J. Galí 2 , P. Teixeira 1 J. Simoes 3 

1 Centro Veterinário de Aveiro, Aveiro, Portugal 

2 Grup Tecnic Veterinari S. L., Gerona, Spain 

3 University of Trás-os-Montes e Alto Douro, Veterinary Science 

Department, Vila Real, Portugal 

Characterisation of pathogenic flora from composite milk (CM) samples 

of subclinical intramammary infected cows is frequently performed, 

instead of quarter milk (QM), in part due to economic reasons. One QM 

or CM samples are also used in cows with only one QM to positive 

California Mastitis Test (CMT). The aim of present work was to compare 

pathogenic flora between QM and CM samples from cows with one 

positive CMT quarter. Twenty Holstein-Friesian cows positive CMT in 

one quarter were randomized from 3 dairy farms. Both QM (positive 

CMT) and CM samples were collected in each cow. Somatic cell count 

(fossmatic) and microbiologic analysis (Plate Count Agar) were 

performed. Staphylococcus aureus were isolated in 35% (7/20), Streptococcus 

uberis in 20% (4/20) and Streptococcus agalactiae in 10% (2/20) 

of cows. Remained cows were affected by others contagious or environmental 

microorganisms, including fungi. Pathogenic flora was observed 

in QM but not in CM in 30% (6/20), in CM but not in QM in 5% (1/20) of 

cows, similar in 15% (3/20) and different in 50% (10/20; P

In dairying, good management practice should not only focus on calving 

and early lactation but also consider the foregoing late lactation, seeking 

to have a maximum of quarters secreting normally (i. e. culture-negative 

and < 100,000 somatic cells/ml foremilk) before dry-off. In order to 

evaluate the impact of the udder health status before dry-off has on the 

subsequent early lactation, quarter foremilk samples from 31 HF cows (n 

= 126 quarters) in Northern Germany (2 nd , 3 rd , 4 th and 5 th lactation) were 

drawn weekly, three times before dry-off and eight times after subsequent 

calving, and analysed cyto-bacteriologically according to IDF 

recommendations. The udder health status as observed before dry-off was 

maintained during the early lactation in approx. 67 %. Of those, the ratio 

of normally-secreting quarters to quarters with secretion impairments was 

almost 1:1. While approx. 12 % of quarters secreting normally before dryoff 

became diseased during early lactation, curing was observed in 

another 21 % of quarters. Although varying in degree between lactations, 

the overall odds ratio analysis showed these factors to be significant (p < 

0,001). Of the quarters with culturally-positive result before dry-off (n = 

30 quarters), bacteriological cure during the dry period occurred in 67 % 

of cases. The same pathogen species re-emerged during early lactation in 

13 % of cases while in 20 %, another pathogen species was diagnosed in 

that period. Regarding odds ratio analysis of the microbiological findings, 

colonisation itself also represents a weak factor of influence, while the 

pathogen type did not play a significant role. Results suggest that the 

udder health status before dry-off is of crucial importance for the 

development of the subsequent early lactation and, with that, the total 

lactational performance of a dairy cow (irrespective of her lactation 

number). The reaction potential of the body seems to account more than 

the species or group of pathogens involved, so that late lactation 

management should not only rely on mere hygiene but also promote 

metabolic homoeostasis (feeding, dry season). 

Key words: dry-off, udder health, early lactation, periparturient 

period 

562 Efficacy of Extended Intramammary Therapy to Treat 

Clinical Mastitis in Lactating Dairy Cows under Field 

Conditions 

V. Kroemker, C. Zinke, J.H. Paduch, D. Klocke, J. Friedrich 

University of Applied Sciences and Arts, Microbiology, Hannover, 

Germany 

Streptococcus uberis and Staphylococcus aureus are important causes 

of mastitis in dairy cows in the entire world. Recent publications 

showed that an extended therapy can be more efficient in combating 

such intramammary infections. The objective of this study was to 

evaluate the effect of an extended intramammary therapy to treat 

moderate and severe mastitis cases in lactating dairy cows under field 

conditions in northern Germany. From December 2005 to August 

2007, a total of 236 clinical mastitis cases (242 quarters) on 10 farms in 

northern Germany were enrolled in the study and randomly assigned 

(blocked by parity and body temperature) to one of three treatment 

groups (intrammammary lincomycin/neomycin 1.5-d (ALK-Control) 

or 5-d (ALL-Treatment 1), and 75 mg cefquinom 5 d (COL-Treatment 

2)). Clinical and microbiological cure rates were evaluated. Treatments 

were initiated before culture results. Cows were observed and 

evaluated on d 1 to 6, 19 and 26. Nine cases in 236 (3.8 %) resulted in 

a lost cow (died or culled). Overall, treatments were not significantly 

different to controls regarding culled cows and clinical cure rate. 

However, when the bacteriological cure rate was evaluated, differences 

were observed. ALL appeared to be the most efficient treatment (43/44 

[97 %] P < 0.01, ALK 24/32 [75 %] and COL 35/50 [70 %]). This was 

due to the fact that ALL yielded increased cure rates towards mastitis 

caused by Streptococcus uberis which was the most frequent micro 

organism. We conclude that in farms with Streptococcus uberis 

mastitis, the 5-d extended lincomycin/neomycin treatment regimen 

was significantly more efficient than the standard 1.5 d treatment. 

Key words: mastitis, extended therapy, Streptococcus uberis 

563 Reasons for Culturally Negative Results of Quarter Foremilk 

Samples from Clinical Bovine Mastitis 

V. Kroemker, C. Zinke, JH. Paduch, D. Klocke 

University of Applied Sciences and Arts, Microbiology, Hannover, 

Germany 

It is well-known that approx. one third of all secretion samples from 

clinical mastitis cases do not yield any cultural bacteriological results. The 


present study was aimed towards analysing the reasons for this 

unsatisfactory situation and eventually proposing some simple 

modifications of the microbiological analysis of milk samples. For this, 

261 quarter foremilk samples from cows with clinical mastitis were reevaluated 

microbiologically after the routine analysis recommended by the 

German Veterinary Association (practically, IDF procedures) yielded no 

results (mastitis pathogen). After repeating this standard diagnosis 

procedure with an inoculum of 0.01 ml, culture-negative samples were 

analysed further. First, inoculum quantity was raised to 0.1 ml. In order to 

free intracellular bacteria, sediment was frozen. Inhibitory substances were 

detected using agar diffusion test with Paenibacillus stearothermophilus 

var. calidolactis. Analysis for Escherichia coli and other coliforms was 

carried out on 3M Petrifilm E. coli/Coliform Count Plates and 3M 

Petrifilm High-Sensitivity Coliform Count Plates, resp. The presence of 

endotoxins of gram-negative bacteria was verified using the limulus test 

while growth of Mycoplasma spp. was done on specific culture media. The 

sediment of samples was analysed both microbiologically on aesculin agar 

and microscopically. Of those 193 samples remaining culture-negative 

after the second standard analysis, mastitis pathogens could be detected in 

a total of 178 samples when the additional procedures were applied. 

Findings related to E. coli and other coliforms (24.4 %), coagulasenegative 

staphylococci (27.5 %), inhibitory substances (17.1 %), 

streptococci (6.7 %), Stapyhlococcus aureus (1.6 %), and mixed infections 

with coagulase-negative staphylococci being involved (7.8 %). Merely 7.3 

% of samples remained bacteriological negative. Increasing the amount of 

inoculum leads to bacteriologically positive findings in 50 % of the 

samples that had been culture-negative before. 

Key words: mastitis, culturally negative, milk samples 

564 Bacteriological Quality of Believed Milk (Case of the Farms 

of the Area of Mitidja) 

D. Guetarni, D. Baazize-ammi, E. Lebres, A. Dechicha, S. Kebbal 

University of Blida, Biology, Algeria, Algeria 

In Algeria, milk is paid neither on bacteriological quality, nor on the rate 

of the somatic cells and even less on the presence of the residues of 

inhibitors on the level of the dairies. The stockbreeders perceive 

premiums on the butyric and proteinic rates. The present study related to 

146 samples of milk of tank taken on the level of the breedings of the 

wilayas of Algiers and Blida (circuit of collection). The evaluation of 

bacteriological quality, on the basis of FAMT, indicating germs of 

hygiene and certain pathogenic germs, i.e. the criteria retained in the 

Algerian legislation, shows that milk is strongly contaminated. The results 

show that 91,78% of the samples present an aerobic flora mésophile total 

> 105 UFC/ml and 80,13% reveal the presence of the total coliformes. 

These flora reflect the conditions of hygiene on the level of the 

exploitations. The high rates of contamination obtained are probably the 

result of a bad hygiene of the draft or a faulty operation of the tank of 

refrigeration. The contamination of milks by Staphylococcus aureus 

accounts for 80,13%. This important contamination can be due to a lack 

of hygiene or the consequence of the infections mammaires because the 

health of worse in our breedings represents a real problem. Escherichia 

coli was highlighted in 17,80% of the analyzed taking away. This germ 

signs a fecal contamination and represents a real danger to the health of 

the consumer. 

Key words: bacteriological quality, believed milk, total aerobic flora, 

Coliformes, Escherichia coli, Staphylococcus aureus 

565 Bacteriological Quality of Believed Milk (Case of the Points of 

Sale of the Area of Mitidja) 

D. Guetarni, D. Baazize-ammi, A. Lebres, A. Dechicha, S. Kebbal 

University of Blida, Biology, Algeria 

This study aimed one hundred raw milk samples from milk shop 

(saling points) of Mitidja region. The valuation of the bacteriological 

quality shows that the milk is highly contaminated. Results showed that 

81% of samples present a total aerobic mesophilic counts > 105 

CFU/ml and total coliforms are present in 86% of milk. These counts 

reflect the hygienic level, the conditions of storage and transport. The 

obviousness of pathogenic agent, Escherichia coli, Staphylococcus 

aureus in 30%, 58% of milks respectively present a real danger for the 

consumer health. Enterococcus are placed in prominent position in 

95% of milks and 13% of milks contained brucella antibodies. 

Key words: bacteriogical quality, raw milk, total mesophilic counts, 

Escherichia coli, Staphylococcus aureus, Enterococcus, Brucella

566 Somatic Cells Count of Milk in Algeria 

S. Kebbal, I. Gharbi, M. Amara, S. Guemra, D. Guetarni 

University of Blida, Veterinary Medicine, Algeria 

For a long time used for the following sanitary and management 

engineering of the milk’s cattle, count cells is been the official criteria of 

the milk’s payment and the control milk. In the present work, we give a 

method of somatic cells’ count to mean of particular counter “COULTER 

COUNTER” which is a new application in Algeria. After calibration of 

the device, an evaluation of its performances has been realised by the 

determination of reliability and repeatability of analysis on samples of 

quarter’s milk classed before microscopic count by the method of Prescott 

& Breed. The results obtained revealed a good correlation (CC = 0, 98) 

with regard to the reference method. the repeatability has showed that the 

coefficient of variation don’t exceed the 5% (3,26%, 2,39% and 3,57%) 

for the cellular concentrations lower than 250 x 10 3 included between 250 

x 10 3 and 750 x 10 3 and superior to 750 x 10 3 Cells/ml respectively. the 

reproducibility showed a coefficient of variation o 0,55%. The 

performances of “COULTER COUNTER” are good and this method of 

measure of cells concentration can serve for the somatic cells count of 

milk with a good precision. 

Key words: Mastitis, concentration, somatic cells, raw milk 

567 Prevalence of Mastitis and Raw Milk Safety in Hamdallaye 

Dairy Cattle (Niger) 

A. Harouna 1 , C. Locatelli 1 , L. Scaccabarozzi 1 , P. Cremonesi 1 , 

V. Bronzo 1 , G. Adamou 1 , P. Boettcher 2 , G. Ruffo 1 , P. Moroni 1 

1 

University of Milan, Milan, Italy 

2 

Institute of Agricultural Biology and Biotechnology (IBBA), Milan, 

Italy 

A study was performed in 2007 in Hamdallaye dairy herds (Niger) to 

establish the prevalence of subclinical mastitis and possible associated 

risks for public health. The studied sites are organized in the cooperative 

“UPROLAIT”, created by Project of Safety of Niamey Periurban 

Farming and Agriculture. Seven herds participated in the study and 572 

quarter milk samples were collected from 143 lactating cows from four 

local breeds: Azawak, Bororo, Djelli and Goudali. Each sample 

underwent bacteriological examination and automated counting of 

somatic cells (SCC). Selected strains (N = 15) of Staphyloccocus aureus 

isolated from the samples were subject to antimicrobial susceptibility 

tests and multiplex PCR to evaluate the potential production of 

enterotoxins A, C, D, G, H, I, J and L. Descriptive statistics of general 

mastitis prevalence, herd prevalence and relationships between bacteria 

and SCC were estimated using SPSS version 15.0. According to surveys 

of dairy farmers, 33% of cows had open sores on their udders and teats. 

Statistically significant differences in prevalence and SCC were 

observed among pathogen species (p < 0,05). General mastitis 

prevalence in Hamdallaye dairy cattle was 28,15% with: S. aureus 

(10,45%); Coagulase Negative Staphylococci (CNS) (15,55%) and 

environmental microrganisms (2,15%) being the most common 

pathogens. Herd prevalence ranged from 2% (Toka binekani) to 18% 

(Gagare) and from 4% (Bartchawal) to 39% (Toka binekani) respectively 

for S. aureus and CNS. The SCC ranged from 103 to 107 

cell/ml of milk. Infections were assigned to four classes, according to 

the major pathogen, and the respective mean SCC ( 2,000x1000cells/ml) and clinical signs. Silver ion was 

infused through intramammary route. Ten quarters of 6 cows were 

infused with 20 ml silver ion solution for consecutive 3 days. Blood 

samples at 0, 1, 2, 3, 5 and 7 days after intra-mammary infusion of silver 

ion were collected for hematological analyses from coccyegeal vein 

using K3 EDTA tube. Milk samples were aseptically collected from the 

infused quarters of each cow at 0, 12, 24, 48, 72, 120 and 168 hrs after 

intra-mammary infusion of silver ion. The clinical signs of mastitis in 

each cow responded differently to intra-mammary infusion of silver ion. 

However, significant reduction in swelling (80%), firmness of 

mammary gland (100%) and clots in milk (86%) was observed in silver 

ion treated quarters. Mean SCS in milk from quarters having clinical 

mastitis was reduced (P

Somatic cell count (SCC) is one of the most common parameters for 

detection of subclinical mastitis. Udder quarters with SCC

virulence factors and virulotypes will be presented. More than 50 

virulotypes were observed when the results were concatenated. Strains 

belonging to the most two frequent virulotypes (9 and 4%) were 

positive for the type 8 capsular antigen and for ClfA/B, FnbA, EbpS, 

SdrC, Spa, HlA/B/D/G, SspA and lukD/E/M, but differed by the 

collagen adhesin and the intercellular adherence factor. Twenty-five 

strains were further compared by the multi-locus sequence analysis 

(MLST) based upon 7 housekeeping genes: 12 strains belonged to the 

clonal complexes 97 and 133, but the other strains belonged to various 

sequence types. Though the clonal complex 133 was related to one the 

most frequent virulotype, the clonal complex 97 comprised strains 

belonging to different virulotypes. Based on this limited number of 

strains tested, the clonal relatedness of S. aureus strains based on 

MLST analysis can not be transposed to the classification into 

virulotypes. Results of similar studies in other countries will help to 

choose the immunogens to incorporate into the vaccines. 

Key words: Mastitis, cattle, S. aureus, virulotyping, Multilocus 

Sequence Analysis 

576 The Effect of Fermentated-herb Extract on Subclinical 

Mastitis in Dairy Cows 

TY. Hur 1 , SJ. Kang 1 , YH. Jung 1 , YJ. Do 1 , CY. Choi 1 , IS. Ryu 1 , 

HS. Kim 1 , YS. Lee 1 , SJ. Park 1 , GH. Suh 2 

1 National Institue of Animal Science, Animal Resources 

Development, Cheonan, South Korea 

2 Chonnam National University, College of Veterinary Medicine, 

Gwangju, South Korea 

Therapeutical potential of fermentated-herb extract (FHE) in bovine 

subclinical mastitis was investigated. The fermentated-herb extract was 

made by the process of aqueous extract of twelve herbs inoculated with 

yeast and probiotic to provide feed additive. Somatic cell count (SCC), 

milk yields and total bacterial count (TBC) were evaluated after fifty 

grams of the FHE fed each cows per day for 4 weeks. The results 

revealed that the FHE reduced the TBC and SCC, and increased milk 

yields. The results suggest that the components of FHE possesses some 

biologically activity in bovine subclinical mastitis. As such, the present 

work substantiates the therapeutic use of fermentated-herb extract and 

also emphasizes on the potential of the commonly available non-toxic 

substance to enhance the cow health. However, more experiments are 

necessary for confirmation of these results. 

Key words: mastitis, fermentated herb, somatic cell count 

577 Prognosticable Fatal Outcomes of Naturally Occurring 

Escherichia coli Mastitis in Dairy Cows – Consumption 

Coagulopathy, Hemoconcentration and Hepatobiliary 

Damage 

K. Mori 1 , S. Hagiwara 2 , H. Okada 3 , H. Nagahata 4 

1 

Engaruchiku NOSAI, Clinic of Farm Animals, Engaru, Japan 

2 

Hokkaido NOSAI, Livestock Insurance, Sapporo, Japan 

3 National Institute of Animal Health, Research Team for Prion 

Diseases, Tsukuba, Japan 

4 

Rakuno Gakuen University, Department of Animal Health, Ebetsu, 

Japan 

Objectives: To determine diagnostic characteristics capable of 

prognosticating the fatal outcomes of naturally occurring Escherichia 

coli mastitis in dairy cows. 

Materials and Methods: Vital signs (temperature, pulse, respiration) 

were measured, and blood tests, including white blood cell (WBC) 

count, antithrombin (AT) activity, platelet (Plt) count, hematocrit (Ht) 

value,-glutamyl transpeptidase (GGT) activity and non-esterified fatty 

acid (NEFA) levels, were conducted for all the cows. These values 

were compared between 7 cows that died due to E. coli mastitis and 17 

cows that survived. Five blood test parameters (AT, Plt, Ht, GGT and 

NEFA) that showed significant differences between the dead and 

surviving cows were used to determine the cut-off values; these 

parameters would also be used as criteria to differentiate between the 

outcomes. The diagnostic criterion to prognosticate the fatal outcome 

was decided using the cut-off values. The practicability of the 

diagnostic criterion was verified by trial use on 51 other clinical cases 

(12 dead and 39 surviving cows). 

Results: All the values measured within 12 hours of onset (day 1) did 

not vary between the dead cows and those that survived. However, 

significant differences in the five parameters (AT, Plt, Ht, GGT and 

NEFA) were observed from the second day (day 2). The diagnostic 

criterion for fatal prognosis was determined using the cut-off values of 

the above-mentioned parameters. The fatal outcomes could be 

predicted if at least two of these parameters were within the ranges 

defined by their cut-off values (AT < 120%, Plt < 15 x 10 4 /ml, Ht 32%, 

GGT 45 IU/l and NEFA > 0.6 mEq/l). The sensitivity, specificity, 

predictive value and likelihood ratio of the diagnostic criterion on day 

2 were 100%, 76%, 64% and 4.25, respectively. Verification of the 

practicability of the diagnostic criterion in the 51 other clinical cases 

examined on day 2 showed the sensitivity, specificity, predictive value 

and likelihood ratio to be 67%, 69%, 40% and 2.16, respectively. 

Conclusions: Consumption coagulopathy (low AT and Plt), 

hemoconcentration (high Ht) and hepatobiliary damage (high GGT and 

NEFA) were determined as the diagnostic characteristics in the fatal 

cases. AT, Plt, Ht, GGT and NEFA values can be used as criteria for the 

prognosis of fatal E. coli mastitis. 

578 Pseudomonas Mastitis: Elimination from Contaminated 

Wash-water Systems by Slightly Acidic Electrolyzed Water 

K. Kawai 1 , M. Kitou 1 , T. Mitamura 2 , I. Uchida 3 , K. Hirose 4 , 

H. Nagahata 5 

1 

Seibu Veterinary Clinical Center, Tokachi NOSAI, Shimizu, Japan 

2 

Ecolofrontier Co., Ltd., Sapporo, Japan 

3 

Hokkaido Research Station, National Institute of Animal Health, 

Sapporo, Japan 

4 

Clinical Research Center, MeiJi Seika Kaisha Ltd., Yokohama, 

Japan 

5 

School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 

Japan 

Objectives of study: Pseudomonas aeruginosa (PA) is usually 

responsible for less than 1% of the mastitis infections in dairy cows. 

The source of infection for PA mastitis is contaminated water used to 

wash the udder or when antibiotics used for intramammary infusion 

have become contaminated. PA mastitis infrequently causes clinical 

mastitis and their spread of infection inflicts damage on the farms. In 

this study, we present effective mastitis control in dairy herd with 

increased incidence of PA mastitis. 

Materials and Methods: 387 dairy cows were milked in the parlor. 

The preventing measures of PA mastitis were started since Oct. 2005. 

Milk samples of clinical cases were obtained for bacteriologic culture, 

and isolated bacteria were identified using standard techniques. Liners, 

jeters, drop hoses and wall of cistern in the parlor were wiped with 

cotton swabs and cultured. Milking procedure was checked and 

changed to the recommended procedure. The chlorine concentration of 

water was increased from 0.05 ppm to 0.5 ppm. Milking equipment and 

floor of parlor were sterilized by 200 ppm sodium hypochlorite. The 

cows infected with PA were isolated and milked at the end. The milk 

samples of all cows were cultured after calving and before dry period. 

Slightly acidic electrolyzed water system (Purester MP240E: Morinaga 

Milk Industry Co., Ltd., Tokyo) was introduced to the parlor and was 

used at milking time since Jan. 2006. 

Results: The occurrences of PA clinical mastitis were 35cows in 2003 

and 45cows in 2004. After implementation of the preventing measures, 

the occurrence of PA clinical cases was 8.3% on all clinical mastitis in 

2005. P. aeruginasa, P. fluorescences and P. putida were isolated from 

environment (eg, liners, jeters, drop hoses, wall of cistern in the parlor 

and reservoir in free stall), and same pathogens were isolated from 

clinical cases and 23 of nonclinical cases. However, after the use of 

slightly acidic electrolyzed water (hypochlorous acid: available 

chlorine, 12 ppm) in 2006, Pseudomonas sp. in the environment, the 

occurrence of PA clinical mastitis and the reservoir of Pseudomonas 

sp. in the nonclinical cases were disappeared. 

Conclusions: The use of slightly acidic electrolyzed water at milking 

time is beneficial in reducing exposure of the cows to the organism and 

it was proved to be effective on prevention of PA mastitis. 

Key words: mastitis, P. aeruginosa, slightly acidic electrolyzed water 

579 Acute Clinically Mastitic Animals in Some Assiut 

Governorate Villages: Diagnosis and Treatment 

M. Sayed 1 , A. Abd El-Rady 2 

1 

Faculty of Veterinary Medicine, Assiut University, Food Hygiene, 

Assiut, Egypt 

2 


Assiut, Egypt 


This investigation was run in some villages in Assiut governorate 

during summer 2007 to diagnose acute clinical mastitis in different 

animal species and to apply different lines of treatment for 

evaluating which treatment line of choice giving cure, aiming to 

another goal by preventing the conversion of acute mastitis towards 

the chronic one which is difficult to be treated and the dairy animal 

will be excluded. Therefore, 2150 animals were clinically examined 

in 5 villages located north to Assiut city, Egypt, including 400 

cows, 950 ewes and 800 she goats, and the incidence of acute 

clinical mastitis was 22.50, 2.63 and 4.63%, respectively. Milk 

samples were collected from all clinically mastitic cows for 

bacteriological examination to identify the causative agents of the 

intra-mammary infection (IMI). It was found the major causative 

agents isolated were Staphylococcus aureus, Streptococcus 

agalactiae, Escherichia coli and Corynebacterium pyogenes, 

causing either single or mixed type of infection. When applying 

different lines of treatment, all diseased animals were classified into 

3 groups: 1 st group received local treatment by intra-mammary 

infusion antibiotic. 2 nd group received systematic treatment by 

intra-muscular (I/M) injection of both antibiotic and antiinflammatory 

drugs. While, 3 rd group received combination of both 

local and systematic treatment lines together. Cure% was achieved 

as 50% for 1st group, 90% for 2 nd group, while 3 rd group gave 

complete cure by 100%. It was noticed that the incidence of acute 

clinical mastitis among examined cows was worrisome and can be 

considered as indicator of the epidemiology of the disease. While, 

spreading of the disease among ewes and she goats was somewhat 

low in comparison with that of cows. In conclusion, combination of 

both local and systematic treatment lines together should be advised 

in treatment of acute clinical mastitis to ensure complete cure. The 

obtained results highlighted the focus towards the spreading of 

acute clinical mastitis among cows lived in some Assiut 

governorate villages. 

580 Efficacy of Extended Therapy of Staphylococcus aureus with 

Intramammary Cefuroxime 

J. Spatz Shelgren, K. Parker, S. McDougall 

Animal Health Centre, Research Division, Hamilton, New Zealand 

Bacteriological cure following antibiotic therapy of Staphylococcus 

aureus during lactation results in cure rates averaging between 25- 

35%, but with a range of between 4 and 92%. Increasing the 

duration of therapy increases cure proportion using beta-lactams 

and lincosamides. However, there no data on the efficacy of 

extended therapy with a cephalosporin is available.Cows (n=61) 

with a SCC of >200,000 were examined with a California Mastitis 

Test (CMT) and those glands with a CMT >1 were aseptically 

sampled for bacteriological culture. Cows (n=34) from which 1 or 

more glands were culture positive for S. aureus (total n=55 glands) 

were blocked by number of glands within cow with S. aureus then 

ranked by SCC before being assigned randomly within blocks of 3 

to no, 3 or 6 intramammary infusions at 12 h intervals with 250 mg 

cefuroxime sodium (Spectrazol Milking Cow, Schering-Plough 

Animal Health, Upper Hutt, New Zealand). Glands were re-sampled 

at 41 and 48 days after the first treatment. Cure was defined as 

having occurred where S. aureus was isolated from neither of the 

both post treatment samples. The probability of cure was modeled 

with main effect being treatment (0, 3 or 6 tubes) on cure at quarter 

level (yes/no). To account for the non-independence of gland within 

cow, the confidence intervals were adjusted by the variance 

inflation factor (pscale; Proc GENMOD, SAS 9.1). A log link 

function was used as exponentiation of the coefficients produces 

relative risks. The cow-level log 10 SCC and milk yield 43 days post 

treatment were analysed using ANOVA with treatment as the main 

effect.The least square mean (95% confidence intervals) cure 

proportions were 0.13 (0.03-0.53), 0.24 (0.10-0.56) and 0.53 (0.32- 

0.87) for glands treated with 0, 3 or 6 tubes respectively. The cure 

proportion tended to be higher following treatment with 6 tubes 

compared to 0 tubes (RR = 4.2 (0.9-19.7); p=0.07) and for 6 tubes 

compared to 3 tubes (RR = 2.2 (0.8-6.0); p=0.11). There was no 

difference in log10 SCC or milk production at the herd test 43 days 

post-treatment among the treatment groups (both p>0.3).It is 

concluded that increasing the duration of therapy with the second 

generation cephalosporin cefuroxime from 0 to 3 and 6 tubes 

resulted in a tendency for a higher bacteriological cure rate in 

glands infected with S. aureus. 


581 Relationship between Somatic Cell Count (SCC) and 

Bacterial Infection of Goat Milk: 1 st Report of Caprine 

Subclinical Mastitis from Iran 

H. Ghasemzadeh-Nava 1 , S. Bokaee 2 , P. Tajik 1 

1 Faculty of Veterinary Medicine, University of Tehran, Department 

of Clinical Sciences, Tehran, Iran 

2 Faculty of Veterinary Medicine, University of Tehran, 

Epidemiology, Tehran, Iran 

The SCC is a good indicator for milk quality and can be implicated as 

a quality character of an animal. An increased SCC is either the 

consequence of an inflammatory process due to the presence of an 

intramammary infection (IMI) or under non-pathological conditions. 

For this reason the SCC of milk represents a sensitive marker of the 

health of the udder and is considered a useful parameter to evaluate the 

relationship between IMI and changes in milk characteristics.The aims 

of our study were (a) to determine the occurrence rate and etiology of 

subclinical mastitis in native meat-type breed goats in Garmsar (a city 

in Semnan province in Iran) and (b) to evaluate threshold of SCC for 

bacteriologically negative and positive goat milk. Goats were 

restrained in a sitting position and the teat end of each half udder was 

scrubbed thoroughly using cotton wool soaked in 70% ethyl alcohol. 

The three first streams were discarded, and 10ml milk samples were 

taken in a sterile tube held horizontally. The tube was capped and 

labeled, then placed on ice and sent to the laboratory for bacteriology 

and SCC within 8 h. Of the 300 milk samples collected, 24 (8%) udder 

halves were bacteriologically positive. The most frequent isolate from 

subclinical cases was cougulase negative staphylococci (90.2% of 

isolates). The other isolate was Pseudomonas aer. In bacteriologically 

positive samples, the lowest and the highest of goat milk SCC were 

1.1x10 6 and 0.5x10 6 cells/ml respectively in which the mean of it was 

0.75x10 6 cells/ml. In bacteriologically negative samples, the lowest 

and the highest of goat milk SCC were 0.7x10 6 and 0.025x10 6 cells/ml 

respectively in which the mean of it was 0.025x10 6 cells/ml. The 

threshold of SCC for negative and positive goat milk was 0.5x10 6 

cells/ml in our study. Isolation of bacteria was associated with an 

increased SCC of goat milk in our study. There is not any report 

pertaining to caprine subclinical mastitis in different breeds from Iran. 

Cougulase negative staphylococci have been considered to be the 

major cause of non-clinical intramammary infections in a number of 

previous investigations. Due to different pathological and nonpathological 

factors contribution to change SCC in milk of goats, a 

complete study to take into consideration of different factors is needed. 

Key words: mastitis, goat, SCC, bacteriology 

582 Comparing the Efficacy of Copan and Delvotest to Detect of 

Antibiotic Residues in Bulk Tank Milk Samples of Garmsar 

Dairy Farms 

M. MOhammadsadegh 1 , S. Bokaii 2 , T. Mosavi 3 , E. Abbaszadeh 4 

1 

Islamic Azad University, Garmsar branch, Large Animal Clinic for 

Theriogenology and Ambulatory Services, Garmsar, Iran 

2 

University of Tehran, Epidemiology, Tehran, Iran 

3 

Islamic Azad University, Garmsar branch, Food Hygiene, Garmsar, 

Iran 

4 

Islamic Azad University, Garmsar branch, Large Animal Clinic for 


Antibiotic residues in milk are from mastitis treatment with 

intramammary infusions of MC ointments an /or with systemic therapy 

in many diseases. Milk antibiotic residues have a hazardous effects on 

milk industry and milk consumer welfare. In order to compare Delvo 

test with Copan kit in field of evaluating antibiotic residue in milk, 100 

bulk tank samples of collected milk were prepared in the milk 

collecting center of Garmsar. Antibiotic residues were analyzed by 

Copan and Delvo test, and total bacterial counts were determined in all 

samples. Results showed that antibiotic residue were present in 19% of 

samples. Nine present of positive samples were found by Delvo test 

and 10% by the two Delvo test and Copan, consequently. Total 

bacterial count in only Delvo test positive samples were 24944/44 – 

1710/35, and in Delvo test and Copan positive were 13160 – 2061/6. 

Statistical analysis showed that substantial agreement of the two test 

was acceptable (kappa = 0.643). It is concluded that Delvo test is a 

significantly susceptible test, at least in present condition of this study. 

Key words: Delvo test. Copan test, antibiotic residues in milk, 

Garmsar

583 An Integrated Approach to Production Process and 

Education in Dairy Cattle Farming: the American Farm 

School 

E. Souglis 1 , G. Arsenos 2 , G. Valergakis 2 , A. Giamoustaris 1 

1 American Farm School, Thessaloniki, Greece 

2 Aristotle University of Thessaloniki, Animal production, 

Thessaloniki, Greece 

The quality of dairy products has become an issue for consumers over 

the last decades due to various outbreaks of food-borne zoonoses. 

Moreover, consumer concern about production methodologies and the 

way that dairy cows are being kept is increasing. Hence, quality 

assurance programmes have been implemented in dairy farms to 

counteract the latter concerns. The objective of this study was to assess 

the quality assurance program implemented by the American Farm 

School of Thessaloniki (AFS). The AFS was chosen because it is a small 

dairy enterprise that first introduced pasteurised milk to Greece in 1935. 

Since then, it has been the industry leader in productivity and quality of 

milk serving also as an education centre in cow herd management and 

milk processing for farmers and students. The AFS dairy herd 

comprised of 124 Holstein cows reared in a freestall barn. All the cows 

were included in this study. Data regarding milk quality parameters, 

disease occurrence and management in relation to health and 

productivity were collected over the last three years. More particularly, 

Somatic cell counts (SCC), Total Bacterial Counts (TBC), fat and 

protein content of milk were measured in individual cows on a monthly 

basis. Milk yield and reasons for culling were assessed every lactation, 

whereas the cost of particular diseases was estimated. The results 

revealed relatively low SCC (mean: 253,628, S.E:19,022), and TBC 

(51,214, S.E: 13,032). Protein content of milk was similar (3.26, 3.28, 

3.21, s.e.d. 0.044) whereas there was a significant increase in fat content 

(3.96, 4.26, 4.09, s.e.d 0.082) over the three lactations, respectively. The 

average replacement rate was 24% and apart from age, culling was 

associated with feet and reproduction problems. It is concluded that 

based on developments within the dairy sector in Greece and neighbour 

countries such practices on dairy farms will be a common practice in the 

near future and the AFS could disseminate such knowledge though its 

educational program. The later is likely to help dairy farmers identifying 

and managing the quality hazards and risks occurring in the production 

process, and hence providing the consumer with more certainty about 

the welfare of dairy cows and the quality their milk at retail. 

584 Apoptosis and Autophagy in Involuting Bovine Mammary 

Gland – the Role of TGF-beta1 and Somatotropic Pathway 

Proteins 

J. Zarzynska 1 , B. Gajkowska 2 , U. Wojewódzka 2 , T. Motyl 3 

1 

Faculty of Veterinary Medicine, Warsaw University of Life Sciences 

- SGGW, Department of Food Hygiene and Public Health, Warsaw, 


2 

Polish Academy of Science, Medical Research Center, Warsaw, 


3 

Faculty of Veterinary Medicine, Warsaw University of Life Sciences 

- SGGW, Department of Physiological Sciences, Warsaw, Poland 

The mechanism of secretory tissue involution in bovine mammary 

gland is still poorly understood. We can find many factors, which 

could regulate this process. It has been shown that expression of 

TGF 2-a1 and its receptors in mammary epithelial cells (MEC) 

increases during mammary gland involution in mouse, goat and sow. 

It has been also shown that TGF 2-a1 is an antiproliferative and 

apoptotic factor in bovine MEC. Our recent study revealed that 

expression of TGF 2-a1 in bovine MEC undergoes complex endocrine 

and auto/paracrine regulation by hormones of the somatotropic axis. 

Similarly to apoptosis, the intensity of autophagy was the highest in 

the dry period, as shown by increased expression of Beclin1 and 

morphological features. Autophagy observed in the involuting 

mammary tissue could be the natural cell defense against transient 

undernourishment and action of apoptogenic peptides (e.g. TGF 2-a1, 

IGFBPs), thus maintaining cellular homeostasis in the dry period. 

Key words: TGF 2-a1, mammary gland, apoptosis, autophagy 

585 A Pilot Study on the Bacteriological Diagnostics of Bovine 

Mastitis Carried Out by Finnish Veterinarians 

H. Simojoki 1 , S. Erkkilä 2 , A. Pitkälä 1 , S. Pyörälä 1 

1 University of Helsinki, Faculty of Vet. Medicine, Department of 

Production Animal Medicine, Helsinki, Finland 

2 Finnish Food Safety Authority Evira, Helsinki, Finland 

Objectives of the study: Bacteriological diagnosis of bovine 

mastitis determines treatment decisions and is important for herd 

health programmes. In Finland, farmers can send milk samples to 

laboratories or in cases of acute mastitis, the practicing veterinarian 

often cultures the milk sample. The aim of this study was to evaluate 

the accuracy of bacteriological diagnostics carried out by practicing 

veterinarians. 

Materials and methods: Thirty veterinarians volunteered to 

participate in the study. Autoclaved milk was inoculated with dilutions 

of mastitis causing bacteria. Samples were sent to the participating 

veterinarians, who were asked to culture the samples according to their 

normal routines. A questionnaire on diagnostic methods and quality 

control systems used by the veterinarians was included. Bacteriological 

diagnoses of the milk samples considered to be correct were A. K. 

oxytoca (Klebsiella sp.); B. Str. uberis and A. pyogenes; C. S. simulans 

(coagulase-negative staphylococci, CNS); and D. E. faecalis 

(Enterococcus sp). The diagnoses were scored as correct, partly correct 

and incorrect. The diagnoses were scored as partly correct if only 

bacterial genus was identified. 

Results: For sample A, the diagnoses were correct in 22 out of 28 

cases, partly correct in 2 and incorrect in 4 cases. For sample B, the 

diagnoses were partly correct in 18 out of 28 cases, and incorrect in 10 

cases. Different Streptococcus species were mentioned and A. 

pyogenes was not recognised. For sample C, diagnoses were correct in 

26 out of 28 cases and partly correct in 2 cases, and for sample D, 

correct in 24 out of 28 cases and incorrect in 4 cases. The two most 

commonly used agars were diagnostic triplate Mastix ® -agar (used by 

64% of the veterinarians) and blood-esculin-agar (21%). Additional 

tests included Gram-staining, rapid coagulase test, tube coagulase-test, 

and various tests for antibiotic sensitivity. Quality control procedures 

were seldom used, but included monitoring the temperature in the 

incubating closet and noting the expiry date of agars. 

Conclusions: Samples with CNS were most frequently diagnosed 

correctly. Sample B, comprising Str. uberis and A. pyogenes, caused 

most problems. Slow growth and small colonies of A. pyogenes 

remained undetected and the Mastix ® -agar was not suitable for 

identifying the various Streptococcus species. Small field laboratories 

run by the veterinarians would need guidelines for implementing 

quality control. 

586 Mycoplasma bovis Outbreak in a Dairy Herd: Consequences 

on Production and Reproduction 

L. Pinho 1 , I. Maia 2 , N. Canada 1 , C. Azevedo 2 , M. Machado 3 , 

E. Silva 1 , G. Thompson 1 , J. Carvalheira 1,4 

1 

Universidade do Porto - ICBAS, Department of Clinical Sciences, 

Porto, Portugal 

2 

Medicina da Produçio Leiteira Veterinária, Tocha, Portugal 

3 

Laboratório Nacional de Investigaçio Veterinária, Bacteriology, 

Vila do Conde, Portugal 

4 

Universidade de Porto, Centro de Investigaçio em Biodiversidade e 

Recursos Genéticos, Vila do Conde, Portugal 

The objective of this report was to document an outbreak of M. bovis in 

dairy cattle. Mastitis was the major pathology, but arthritis and pneumonia 

were also observed, including a decrease in reproductive efficiency. The 

outbreak occurred in a dairy farm with 160 lactating cows in northwest 

Portugal. To determine the cause of the mastitis outbreak, individual and 

bulk tank milk samples were obtained between September 2006 and April 

2007. Milk samples were inoculated in modified Hayflick Agar and 

enrichment solution. Standard cultures for other bacteria detection were 

also performed. Mycoplasma species were identified according to 

biochemical characteristics and PCR. M. bovis was isolated from 13 of the 

48 individual milk samples collected. At the peak of the outbreak, the rate 

of clinical mastitis exceeded 10 cases per month, with the majority of 

cows presenting elevated SCC and decreased milk yield that persisted 

even in clinically recovered animals. Multiple quarters were affected with 

little or no response to antibiotic therapy. Atrophy of affected quarters or 

complete destruction of the gland, was also observed. Prophylactic 

measures included segregation and culling of the positive animals, 

introduction of iodine based pre and post-dipping and the use of single 

towels. Treatment was changed to single disposable intramammary tubes 

instead of systemic antibiotics given by intramammary route. The 


eproductive data showed that cows found positive to mycoplasma had 

lower reproductive efficiency. The percentage of cows with 3 or more AI 

per conception was higher (82%) for infected cows. The same pattern was 

observed on other parameters such as days-open and metritis incidence. 

The rate of pneumonic infections was highest after the onset of the 

outbreak. Standard treatments were inefficient. Arthritis was also 

diagnosed in some cows showing enlarged joints and lameness. There 

was an increase of SCC between September and November in positive 

cows. During those 3 months 67%, 83% and 100% of the infected cows 

had SCC above 200,000 cells/ml with an average higher than 1,200,000 

cells/ml. The production level of the herd decreased from 26.5 kg in 

August to 22.6 kg of milk/day/cow in November. This report confirmed 

for the first time the association of M. bovis with clinical and subclinical 

mastitis in dairy cattle in Portugal. These results suggest that M. bovis 

should be considered in the differential diagnosis of mastitis in Portuguese 

dairy farms. 

Key words: mycoplasma bovis, dairy cattle, mastitis, arthritis, 

pneumonia 

587 Bulk Milk Tank Staphylococcus aureus UCF Count as 

Predictive Datum for Bovine Herd’s Infection Level 

G. Varisco 1 , L. Bertocchi 1 , N. Contessi 2 , M. Tranquillo 1 , G. Bolzoni 1 

1 Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia 

Romagna Brescia, Brescia, Italy 

2 Centro Miglioramento Latte, Brescia, Italy 

Aim of this work is to verify the predictivity of bulk milk tank 

(BMT) analysis as simple tool for epidemiological surveillance system on 

herd’s Staphylococcus aureus infection level. The comparison between 

BMT analysis results and single cow milk (SCM) analysis results was 

investigated to verify the relationship between BMT Staph. aureus UFC 

value and dairy herds infection rate. During 2005, 47.208 cow milk 

samples were analysed (7.166 positive, 38.530 negative and 1.512 to 

much polluted)) of 31 dairy herds (approximately 4.000 cows were 

checked). Staph. aureus count in BMT (UFC) was found to 

decrease during time, due to dairy Staph. aureus infection rate reduction 

at cow level. Statistical analysis shows that within herds the prevalence is 

1.28 grater (exp 0.25) every one UFC log unit (p 

7 log) were removed by the bedding treatment. 

Conclusions: The results indicate that applying DryMaxx bedding 

treatment to the bacterial suspension facilitated the removal of the bacterial 

suspension from the cow mat. We suggest applying DeLaval bedding 

treatment to DeLaval cow mats, letting the bedding treatment remain on 

the cow mat for at least 30 minutes, and then sweeping off the bedding 

treatment and removing it from potential future contact with the cow. 

589 Evaluating the Teat Condition Performances of a New Teat 

Spray in an Automatic Milking Environment 

X. Goossens 1 , L. Bommelé 2 , W. Ingalls 3 , T. Hemling 3 

1 

DeLaval, PP Milk Quality & Animal Health, Gent, Belgium 

2 

Ghent University, Biocenter Agri-Vet, Melle, Belgium 

3 

DeLaval, PP Milk Quality & Animal Health, Kansas City, United 

States 

Automatic milking is a growing practice in dairy farming. In an automatic 

milking system the cows are milked more frequently and the variation in 

the interval between milkings is greater than with conventional milking. 

As the automatic milking creates a new and different management and 

environment with different challenges for the teats, new teat dips with the 

objective to maintain good teat condition when sprayed in an automatic 

milking system should be tested in the system itself. Results abstracted 

from conventional milking parlours cannot be the only basis to evaluate 

teat dips in this new way of milking. Teat condition studies done in 

conventional milking parlours, use a split udder or split herd design. In 

the conventional milking parlours this dipping or spraying is done by 

human interaction and can be done selectively with the right product on 

the correct teats. In automatic milking systems, teat spraying is done 

automatically. Technically it is not possible to spray the teats of 

different cows with different products. And because of the spraying it 

is not possible to spray only two teats of each cow without hitting the 

two control teats. This makes it impossible to do a side by side control 

trial in a robotic system. The objective of this study was to evaluate a 

new teat spray for an automatic milking system in a commercial dairy 

farm environment with a positive control. To evaluate teat condition 

performance of a new peroxide teat spray (DeLaval Prima) in an 

automatic milking system we made use of a switch back design 

(Rasmussen et al. 2001). Control product was Delaval Proactive Plus, a 

teat dip with well know teat condition performance. Before the start of 

the trial, the teats had already been sprayed for a long period with the 

control product. During the trial period of 18 weeks, the experimental 

product and the control product were switched every 6 weeks. At the start 

and every two weeks during the trial period, teat skin condition, teat end 

condition and orifice thickness were scored by the same trained person 

using an ordinal scale. Every two weeks individual SCCs were recorded. 

Weather data were gathered throughout the trial. The switch back design 

should help to evaluate the teat condition performances of a new teat 

spray in an automatic milking environment with one robot. 

590 Prevalence of Intramammary Infection in Pregnant Heifers 

M. Vasil, J. Bires 

University of Veterinary Medicine, Animal Husbandry and Veterinary 

Science, Kosice, Slovakia 

Examination carried out in 3 farms on 110 heifers. Clinical 

examination of the udder and bacteriological tests of quarter secretion

were performed between the 4 th and 2 nd week before parturition. The 

sings of clinical mastitis were found in udder of 22 heifers (20%). 

Intramammary infections caused by 15 species of microorganisms 

were found in 140 quarters (32.1%) of 78 pregnant heifers (70.9%). 

Arcanobacterium pyogenes, Streptococcus agalactiae, Streptococcus 

uberis, Streptococcus dysgalactiae, Escherichia coli, Staphylococcus 

aureus and coagulase-negative staphylococci (S. hyicus, S.. 

epidermidis, S. simulans, S. cohnii, S. xylosus, S. intermedius, S. 

haemolyticus) were most frequently isolated. 

Key words: mastitis, heifers, intramammary infection 

591 Concentration of Ascorbic Acid in Blood of Cows Affected 

with Clinical Mastitis before and after Treatment Using 

Cobactan 

M. Kleczkowski, W. Klucinski, T. Jakubowski, K. Dembele 


Clinical Sciences, Warsaw, Poland 

The aim of the investigations was evaluation of AA concentration in 

the cows serum with clinical mastitis caused by Staphylococcus aureus 

before and after treatment using Cobactan. The experiment was 

conducted on 30 cows, Black-White breed, at the age of 3-8 years. The 

animals were divided into two groups (A-experimental and B-control). 

All cows were subjected clinical examination, evaluation of 

macroscopic changes of milk, and evaluation of somatic cell count. 

Milk samples, before treatment, were taken simultaneously for 

identifying pathogenic micro-organisms. All cows from group A were 

given Cobactan LC during three days, at 24-hour intervals. All cows 

from group A were also treated intramusculary with Cobactan once 

daily, for 3 days. Blood samples from all cows (group A and B) were 

collected from the external jugular vein 9 times: 1 st -before giving a 

drug, 2 nd -6 hours (h) from the first dose of drug, 3 rd - 6 h from the 

second dose of drug, 4 th -6 h, 5 th - 24 h, 6 th -48 h, 7 th -72 h, 8 th -96 h and 

9 th -120 hours from the third dose of drug. AA concentration was 

measured by the hydrazine method. Somatic cell count in the milk 

cows from group A also was higher (1.81 million) than in milk of cows 

from group B (0.56 million).During the bacteriological examination of 

milk samples Staphylococcus aureus was isolated. Results of 

experiment show, that average serum AA concentration in group 

experimental A was 48.99 µmol/dm 3 , but in group control B 63 

µmol/dm 3 .The results of clinical examination of the mammary gland, 

organoleptic evaluation of mammary gland secretion, number of 

somatic cells, and bacteriological examinations of milk indicated a 

clinical form of mastitis caused by Staphylococcus aureus in cows 

from group A. It confirm of decrease antioxidant protection potential in 

blood of cows with clinical inflammation of mammary gland. 

Key words: cow, mastitis, ascorbic acid 

592 Glutathione Peroxidase Activity in Blood of Cows Affected 

with Subclinical Mastitis before and after Treatment Using 

Cepravin TM Dry Cow 

T. Jakubowski, W. Klucinski, M. Kleczkowski, K. Dembele 

Warsaw University of Life Sciences, Clinical Sciences, Warsaw, 


The aim of the investigations was evaluation of GSH-PX activity in the 

whole blood of cows with subclinical mastitis caused by Streptococcus 

agalactiae before and after treatment using Cepravin TM Dry Cow.The 

experiment was conducted on 24 cows, Black-White breed, at the age 

of 4-7 years. The animals were divided into two groups (Aexperimental 

and B- control, 12 animals in each). All cows were 

subjected to an initial clinical examination, including the mammary 

gland, evaluation of macroscopic changes, and evaluation of somatic 

cell count in milk. Milk samples, before treatment, were taken 

simultaneously for identifying pathogenic micro-organisms. However, 

before treatment of 12 cows the somatic cell count was estimated with 

the microscopic method. All cows from group A were given Cepravin 

TM Dry Cow. The drug was given once intramammary to each quarter 

separately during 1 st day of dry period. Blood samples from all cows 

(group A and B) were collected from the external jugular vein 9 times: 

1 st -before giving a drug, 2 nd -6 hours (h) from treatment, 3 rd - 24, 4 th -48, 

5 th -72, 6 th -96, 7 th -120, 8 th -144 and 9 th -168 h from treatment. GSH-PX 

activity in the whole blood of cows was measured by the GSH-PX- 

Randox test.The examinations indicate on subclinical mastitis only in 

cows from group A. Somatic cell count in the mammary gland 

secretion 12 cows from group A before treatment was higher (660 

000/ml) than in the milk of 12 cows from group B (107 000/ml).During 

the bacteriological examination of milk Streptococcus agalactiae was 

isolated. Results of experiment show, that average GSH-PX activity in 

the whole blood in group experimental A was 42.1 U/g Hb, but in 

group control B 33.1 U/g Hb. The higher one was in the blood of cows 

from group A-before treatment (51.1 U/g Hb). It confirms an increase 

of enzymatic antioxidant protection in blood of cows with subclinical 

inflammation of mammary gland 

Key words: cows, glutathione peroxidase, antioxidants, mastitis 

593 Etiology of Bovine Mastitis in the Municipality of Patzcuaro, 

Michoacan, Mexico 

C. Bedolla 1 , A. Mejia 1 , S. Renteria 1 , J. Conejo 1 , H. Castaneda 2 , 

W. Wolter 3 , M. Kivaria 4 

1 

Universidad de Michoacan, Department for Infectious Diseases - 

Epidemiology; Faculty of Veterinary Medicine, Morelia, Mexico 

2 

Universidad de Guadalajara, Department for Infectious Diseases - 

Epidemiology; Faculty of Veterinary Medicine, Guadalajara, 

Mexico 

3 

Landesbetrieb Hessiches Landeslabor, Department of Veterinary 

Medicine, Giessen, Germany 

4 

Central Veterinary Laboratory, Department of Microbiology, Dar es 

Salaam, Tanzania 

The objective of this study was to determine the bovine mastitis 

etiology in the municipality of Patzcuaro, Michoacan, Mexico. The 

research was made from September to November 2007 in Patzcuaro, 

Michoacan. Eight milk cattle barns of Holstein Friesian cows were 

sampled, which count on 10 cows in average and which they are raised 

in small scale cattle barns production system. A total of 347 milk 

samples were collected aseptically in sterile tubes of single udder 

quarters of the mammary gland in 88 lactating cows and transported to 

the laboratory to low temperature (4° C). The samples were inoculated 

in both, blood agar with esculin and McConkey agar, incubated at 37 

oC, and observed 24 h and 48 h after inoculation to assess colony 

growth. Staphylococci isolations were reinoculated in agar with sheep 

blood, isolated, in order to observe colonial morphology, hemolysis as 

well as Gram stain, catalase tests, coagulase, manitol and gelatinase. 

The Gram negative pathogens were identified accord to colonial 

morphology, and biochemical tests. From the results of the 

bacteriological examination of the milk samples, 64 (18.44%) 

corresponded to Staphylococcus aureus, 85 (24.49%) to coagulase 

negative staphylococci, 24 (6.91%) to esculin positive streptococci, 2 

(0.57%) to Escherichia coli, whereas 122 (35.46%) corresponds to 

other pathogenic bacteria. We concluded that coagulase negative 

staphylococci were the main pathogen agents found of the bovine 

mastitis, by self the Staphylococcus aureus, was the main causal agent 

of the contagious bovine mastitis in this study. 

Key words: bovine mastitis, small scale barns, S. aureus, clinical 

mastitis 

594 Origin of Clinical and Subclinical Mastitis in Dairy Herd’s of 

the Federal State Jalisco, Mexico. 

H. Castaneda 1 , S. Jäger 2 , W. Wolter 2 , M. Zschöck 2 , 

M. Castaneda 1 , G. Perez 1 , J. Serratos 3 

1 

Universidad de Guadalajara, Department of Veterinary Medicine, 

Guadalajara, Mexico 

2 

Landesbetrieb Hessiches Landeslabor, Department of Veterinary 

Medicine, Giessen, Germany 

3 

Instituto Tecnologico de Tlajomulco, Depto de Biotecnologia, 

Tlajomulco, Mexico 

Aim of the present work was to prove the occurrence of subclinic and 

clinic disturbances of udder health in 33 herds of dairy cattle in Jalisco, 

Mexico. 1996 (66.9%) out of 2937 udder quarters examined by means 

of California Mastitis Test, showed a positive reaction, 1087 (37%) out 

of these reactions were from clearly up to significantly positive 

reactions. Compared to the bacteriological examinations the prevalence 

for subclinical mastitis came up to 43.7%. On the other hand clinical 

mastitis could be proved in 2.5%. In 53.8% of the examined quarter 

milk samples there was no bacteriological pathogen content. From the 

rest of the samples we could isolate CNS (coagulase negative 

staphylococci) (15.4%), Corynebacterium spp (13.9%), Streptococcus 

agalactiae (6.6%), Staphylococcus aureus (5.8%), coliform pathogens 


(3.6%) and others (Bacillus spp., Nocardia spp, Candida spp.) (1.7%). 

These results demonstrate a significant share of minor pathogens beside 

contagious mastitis pathogens as S. aureus and S. agalactiae in mastitis 

incidents in Jalisco, Mexico. 

Key words: clinical and subclinical mastitis, bacterial pathogens, dairy 

cattle, Staphylococcus aureus, Streptococcus agalactiae 

595 Evaluating Post-Milking Teat Dip Efficacy Using Somatic 

Cell Count Data 

A. Bradley 1,2 , J. Breen 1,2 , P. Janowicz 3 , M. McKinzie 3 , T. Hemling 3 

1 

University of Bristol, School of Veterinary Science, Bristol, United 

Kingdom 

2 

Quality Milk Management Services Ltd., Wells, United Kingdom 

3 

DeLaval, Kansas City, United States 

Objectives: Post milking teat disinfectants are traditionally assessed 

by identifying new intramammary infections (IMI) using regular 

individual quarter bacteriology. Somatic cell counts (SCC) are 

recognised as a proxy for IMI; quarters can be classified as “infected” 

or “uninfected” and new infections defined by movements across 

defined thresholds. The aim of this study was to compare the efficacy of 

two post milking teat disinfectants using both bacteriology and somatic 

cell counts, and to assess the utility of these two approaches. 

Materials and Methods: Two teat dip formulations (DeLaval Prima 

(0.5% hydrogen peroxide) and Hamra Blue (0.4% chlorhexidine)) were 

compared on three farms in the UK. Four hundred and fourteen cows 

were randomly allocated (ratio 1:1) to be dipped post milking with one 

of the two formulations. Cows were recruited over a 20 week period; 

377 cows were eligible for inclusion in the subsequent analyses. Quarter 

milk samples were collected for bacteriology and SCC from all cows at 

enrolment and on completion of the study. During the study, quarters 

were sampled for SCC every 14 days. Bacteriology to confirm new IMIs 

was performed when quarters experienced a rise in SCC from below to 

above 100,000 cells/ml. All cases of clinical mastitis were sampled 

during the study in order to determine the cause of IMI. New IMIs were 

defined using various criteria; efficacy was evaluated using statistical 

protocols recommended by the National Mastitis Council (NMC). 

Results: Efficacy of the two products was compared; new IMIs were 

defined by movements across the 100,000 cells/ml threshold alone, and 

in combination with supporting bacteriology. Efficacy was also 

evaluated by comparing clinical mastitis incidence and the change in 

prevalence of IMI between entry and exit of quarters from the study. 

With the exception of the change in prevalence of infection between 

entry and exit from the study (due to a difference in minor pathogen 

IMI), none of the measures demonstrated a significant difference 

between the products. Analysis by various statistical protocols 

recommended by the NMC showed the experimental product (DeLaval 

Prima) was equivalent to the control product (Hamra Blue). 

Conclusion: The use of SCCs, with/without the judicious use of 

bacteriology offers a viable, cost effective alternative to the extensive 

use of bacteriology alone in assessing post-milking teat dip efficacy. 

596 The Effectiveness of Cefalosporine+Kanamicine and 

Ampiciline Gentamicine + Dexametazone in the Treatment of 

Clinical Mastitis in the Livestock Bovine Milk in Mexico 

M. Posadas, L. Matus, O. Blanco, B. Pena 

Facultad de Medicina Veterinaria y Zootecnia-UNAM, Animal 

production, México DF, Mexico 

The present study was carried in Tizayuca Hidalgo, Mexico during July 

to September in order to determining the effectiveness of the treatment 

with the combination cefalexine monohydrate, kanamicine sulfate and 

ampiciline - gentamicine + dexametazone. We administered by the 

intrammary road for the solution of problems of clinical mastitis in 

bovine producing of milk. The sample was took in a population of 50 

animals diagnosed with clinical mastitis and that they presented 

alterations in the milk besides that they obtained a grade 2 in test of 

California (CMT), which were integrated in two groups of 25 glands 

distributed randomly.The control group (A) it was conformed by 25 

glands affected which was administered cefalexine monohydrate + 

kanamicine sulfate during 2 days for a intramamary road.The witness 

group (B) was conformed by 21 affected glands, to which was 

administered: ampiciline - gentamicine + dexametazone for a 

intramamary road with it forms to their medical prescription. They took 

samples of milk of the glands affected to carry out the bacteriological 


exam and to know the etiology.The isolated agents of the sample of milk 

of the two groups “A” and “B” was the following ones: Streptococcus 

agalactiae, Staphtlococcus aureus, Corynebacterium pseudotuberculosis, 

Pasterella spp., Moraxella spp., Eschericha coli, Staphylococcus 

epidermidis, Staphylococcus, saprophyticus, Serratia spp., Streptococcus 

uberis, Citrobacter spp., coagulase negative Staphylococci, 

Actinobacillus salpingitidis, Staphylococcus intermedius and Aeromona 

hydrophila. The results demonstrated that the combination cefalexine 

monohydrate and kanamicine sulfate, it had an effectiveness of 88% to 

control the presentation of clinical mastitis of the group “A” in 

comparison with the group “B” that was of 66%. In the statistical analysis 

they were not significant differences (P=.065>0.05). As for the 

presentation of alterations in the milk, the animals of the group “A” they 

recovered in 86% while the group “B” was of 76%. With regard to the 

test of California for the group “A” 82% of the glands didn’t present 

reaction some after the treatment while in the group “B” was of 68%, 

what allows to deduce that the cefalexine monohydrate and kanamicine 

sulfate is a highly advisable product for the treatment of clinical mastitis. 

597 Effect of Anhydrous Cephalonium-Based Drug Indicated for 

Bovine Mastitis Treatment during the Dry Period on 

Phagocytosis Rates 

C. Batista, M. Blagitz, M. Azedo, C. Stricagnolo, 

M.C. Sucupira, A.M. Della Libera 

Faculdade de Medicina Veterinaria da Universidade de Sao Paulo, 


The dry period is considered a very vulnerable phase of lactation to 

mastitis and the phagocytosis mechanisms are fundamental for this 

period. The antibiotic therapy used for the treatment of dry cows acts 

on the elimination of existing intramammary infections, avoiding new 

infections and decreasing its incidence on the next lactation. An 

efficient drug must contain an adequate antimicrobial component in 

harmony with other eventual components and must be harmless to the 

organism’s defense mechanisms. Twenty-one paired samples, negative 

to bacteriological tests, were used to evaluate the influence of 

Cepravin ® , an anhydrous cephalonium-based drug indicated for the 

treatment of mastitis during the dry period, on the phagocytic function 

of milk leukocytes. In order to evaluate the cellular activity, leukocytes 

were isolated in adequate quality (viability) and quantity 

(concentration) and submitted to in vitro phagocytosis test with 

Zymosan (Saccaromyces cerevisae cell wall), after exposition to a 

solution containing the drug. The average phagocytosis rate for cells 

submitted to the treatment (64.43% ± 15.10) did not differ in relation to 

the average phagocytosis rate for the control group (58,33% ± 12,00). 

In addition, the coefficient of variation of phagocytosis rate obtained 

from cells submitted to the treatment (23.44) was not different from the 

coeficient found in control samples (20.57). The results suggest that the 

treatment, besides the antibiotic activity, does not interfere in the 

phagocytic function of milk leukocytes, which is fundamental for the 

defense of mammary gland, mainly during the dry period. 

Key words: mastitis, treatment, phagocytosis, dry period 

598 Microbiological Evaluation, Somatic Cell and Differential 

Leukocytes Count in Relation to California Mastitis Test in 

Sante In’s Ewes 

M. Blagitz 1 , F. Souza 1 , V. Gomez 1 , C. Batista 1 , M. Azedo 1 , 

C. Strincagnolo 1 , N. Benites 2 , P. Melville 2 , M.C. Sucupira 1 , 

A. M. Della Libera 1 

1 Faculty of Veterinary Medicine and Animal Husbandry FMVZ-USP, 


2 Faculdade de Medicina Veterinaria da Universidade de Sao Paulo, 

Preventive Medicine and Public Health, Sao ppaulo, Brazil 

To evaluate the variation in microbiological profile, total somatic cell 

(SCC) and differential leukocytes counts (DSCC) in milk of Santa Ins 

ewes according to CMT scores, 170 milk samples were examined for 

microscopic somatic cell count (SCC), differential leukocytes count 

(DSCC), microbiological examination and their relationship with 

California mastitis test (CMT). On the 24.21% bacteriologically 

positive samples, 86.96% were coagulase negative staphylococci 

(CNS), and 13.04% were Streptococcus dysgalactiae. The mean (± 

Standard deviation) logarithm SCC of uninfected sheep’s gland was 

5.01 (±0.87) and 5.93 (±0.94) (p

30.00%, 58.97% and 58.33% in negative, 1, 2, and 3 CMT scores 

respectively. The proportion of glands considered positive for CMT 1, 

2 and 3 was 29.41%, 23.53% and 7.65% correspondingly. The results 

showed a higher somatic cell count on higher CMT scores, but no 

significant difference on 2 and 3 scores, thus the means (± Standard 

deviation) logarithm SCC according to CMT scores were 4.44 (+0.57), 

5.21 (±0.46), 6.35 (±0.49), 6.52 (±0.73). Besides this, a positive and 

significant correlation (r = 0.822, p

602 Coagulase Gene Polymorphism of Staphylococcus aureus 

Isolated from Subclinical Bovine Mastitis 

O. Aslantas 1 , C. Demir 2 , H. Turutoglu 2 , Z. Cantekin 1 , Y. Ergun 3 , 

G. Dogruer 3 

1 Mustafa Kemal University, Microbiology, Antakya, Turkey 

2 Mehmet Akif Ersoy University, Microbiology, Burdur, Turkey 

3 Mustafa Kemal University, Reproducción, Antakya, Turkey 

This study was conducted to investigate the coagulase gene 

polymorphism of Staphylococcus aureus isolated from subclinical 

bovine mastitic milk samples from 3 different Turkish cities (Hatay, 

Gaziantep, and Burdur). Amplification of the coagulase gene from 80 

S. aureus isolates produced 5 different polymerase chain reaction 

(PCR) products; 79 isolates showed only 1 amplicon, and 1 isolate 

showed 2 amplicons. The isolates were grouped into 9 genotypes by 

analysing the restriction fragment length polymorphism (RFLP) of the 

gene; the 2 most common genotypes accounted for 73.8% of the 

isolates. The results indicated that many coagulase gene genotypes are 

present in the studied regions and only 1 or 2 genotypes predominated. 

Key words: Staphylococcus aureus, RFLP-PCR, mastitis 

603 Variation of the Cow Milk Quarter-level Somatic Cell Count 

and Lactose Concentration in Relation to Blood Urea, 

Complement and Hematological Parameters 

I. Lusis 1 , V. Antane 2 

1 Faculty of Veterinary Medicine, Latvia University of Agriculture, 

Institute of Food and Animal Hygiene, Jelgava, Latvia 

2 Faculty of Veterinary Medicine, Latvia University of Agriculture, 

Clinical institute, Jelgava, Latvia 

Introduction: Immune system status in the cows body is essential to keep 

udder healthy and free from mastitis pathogens. Activation of the immune 

response is characterized by increase in cell activity, release of mediators 

and increased number of cells in tissues, body fluids, secretions. It is 

necessary to differentiate between adequate or effective somatic cell count 

(SCC) increase in milk and such as long lasting, excessive or pathological 

one. Subclinical mastitis is the main cause for SCC increase. 

The aim: Evaluate correlation among quarter-level SCC, lactose 

content, bacteria in milk and cow blood urea nitrogen, complement C3, 

C4 concentrations as well as hematological parameters. 

Material and methods: Group of 20 cows was selected in a 400-dairy 

cow herd of teaching and research farm Vecauce.Fat, protein, lactose 

concentration and SCC were estimated in the quarter milk samples by 

Milkoscan and Somacount analysers. Milk from each quarter was tested 

bacteriologically with direct culture on the blood agar. Mammary 

infections were categorized in three groups without infection (score 0), 

coagulase negative staphylococci (score 1), major pathogens 

(Staphylococcus aureus, Streptococcus agalactiae, Streptococcus 

uberis) (score 5). Transformation of the quarter level data to cow level 

ones was performed. Arithmetic mean for LgSCC, lactose, milk protein, 

fat concentrations and sum for bacterial infection scores were used. 

Concentration of the comlement factors C3 and C4 in citrated venous 

blood samples were detected by immuneturbidimetry. Correlation 

coefficients among all parameters were calculated by software STATA 

9.0 and showed in results only significant ones (p

Materials and methods: Strains tested were French isolates of 

Escherichia coli, Staphylococcus aureus and Streptococcus uberis. 

Time-kill curves were determined either in Mueller-Hinton broth or in 

UHT sterilized milk over 24 hours for both antimicrobials alone and in 

combination. 

Results: A synergistic bactericidal effect of cefalexin and ciprofloxacin 

was observed on Escherichia coli and Staphylococcus aureus. 

Bactericidal action of the combination against Streptococcus uberis 

reflected mainly effect of cefalexin (combination considered as 

indifferent against this pathogen). 

Conclusion: In vitro interaction observed here may be of clinical 

relevance to improve mastitis recovery in cattle. 

Key words: cefalexin, ciprofloxacin, enrofloxacin, mastitis, synergy 

607 Keratin Plug Presence in Heifers 60 Days before Calving and 

its Relation with Intramammary Infections 

L. Jimienez 1 , R. Timon 1 , J. Marco 2 

1 

Servet Talavera, Dairy Milk Quality Centre, Talavera de la Reina, 

Spain 

2 

Public Health Laboratory, Health Department, País Vasco, Spain 

Objectives: To study the prevalence of intramammary infections 

(IMI), their nature and their relation with the presence of the keratin 

plug, determining the: 1. Prevalence of IMI; 2. Presence of the keratin 

plug (KP); 3. Microbiology of the samples. 

Materials and Methods: The study included 158 Holstein heifers 

from two intensive farms located in central Spain. Milk samples were 

taken from each quarter 60 days before calving, according to the 

National Mastitis Council (NMC). At the moment of sample collection 

the presence of the keratin plug was assessed. Microbiological testing 

was performed according to NMC procedures. 

Results: The prevalence of isolated micro-organisms showed a 

36,7% of IMM with coagulase negative staphylococci (CNS) found 

in 75% of the isolates, Arcanobacterium spp in 6%, other Gram 

positive in 10%, other Gram negatives in 7%, 6% of yeast and 1% of 

mixed culture. 

The distribution of CNS isolated from milk samples from heifers 60 

days before calving was the following: Staph. choromogenes the most 

prevalent 66,4%), followed by Staph. sciuri (8,8%) and, Staph. lentus 

(5,2%). Other isolated species were: Staph. Auricularis, Staph. capitis, 

Staph. caprae, Staph. cohnii cohnii, Staph. epidermidis, Staph. 

equorum, Staph. gallinarum, Staph. haemolyticus, Staph. hominis, %), 

Staph. simulans, Staph. warneri, Staph. xylosus. The distribution of 

CNS species isolated (173 strains) from milk samples obtained from 

heifers 60 days before calving. The criterion of novobiocin resistancesensitivity 

to determine pathogenicity was used, with 90% of the strains 

being sensitive to novobiocin (CNNSS), and 10% of the strains 

resistant to novobiocin (CNNRS). The presence of the KP was assessed 

in a total number of 628 teats, and it was only found in 306 teats 

(48,7%). The stratified IMI study depending on the presence or not of 

the KP showed that heifers without KP had 59 % quarters with IMI, 

while quarters with KP only had 13 % of IMI (P

2 Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of 

Bern, Berne, Switzerland 

Objectives of study: The survey was conducted to estimate the 

prevalence of mastitis pathogens in Swiss dairy cows. The correlation 

between Somatic Cell Count (SCC) and the different pathogens was 

investigated. 

Materials and Methods: The selection procedure was a random onestep-cluster-sampling 

with stratification according to herd size. Quarter 

foremilk samples (QFM) were taken from each cow in lactation of the 

47 herds for microbiological analysis and SCC determination. QFM 

samples were considered as positive if > 1 colony was isolated from 

10µl milk. A cow was considered positive when at least one quarter 

yielded a positive result. All isolated streptococcus spp. were further 

differentiated. 

Results: The cow prevalence of the most important pathogens of 

subclinical mastitis ranged from 0.5% to 62.7% (Tab.1). Of the isolated 

streptococcus spp. 21.5% were S. uberis, 21.7% S. dysgalactiae, 1.3% 

enterococcus and 48% were diagnosed as probably apathogenic 

streptoococci (lactococcus or rare types of enterococcus). The quarters 

infected by S. aureus, streptococcus spp., coagulase-negative 

staphylococci (CNS) and C. bovis had a significantly higher SCC than 

QFM with a negative result (Wilcoxon rank sum test, pMIC 90 (time during which the concentrations exceed the 

MIC 90).Milk maximal concentration (C max) was 7.6 µg/mL (16.7h 

post-1 rst adm) and 7.7 µg/mL (14h post-2 nd adm). Milk elimination 

half-lives 50 were 128.1h and 157.9h after 1st and 2 nd administrations. 

The mean milk AUC 0-24h was 154.2 µg.h/mL. Plasma concentrations 

were lower. The production level had a major effect on the time during 

which antimicrobial concentrations in milk exceeded the MIC 90 

(T>MIC 90), being longer in quarters of low-producing cows. The 

AUC 0-24h in milk was higher in the low production group, although the 

ratio AUC 0-24h in milk/AUC 0-24h in plasma (F milk) may indicate that 

antimicrobial passage is higher in quarters of high-producing cows. 

Infection presence in the quarters had a significant effect (p

hemolysis, catalase test, glucose and manitol fermentation in aerobiosis 

and anaerobiosis and acetoin production by Voges-Proskauer test, 

antibiotic sensitivity test. Organisms were identified as Staphylococcus 

aureus with in vivo sensitivity for gentamicin. All 14 mastitis cases 

treated with 300 mg q 12h of gentamicin, after complete milking from 

the affected udder quarter, five minutes after intramuscular injection of 

20UI ocytoxin, for five days. It was administred 3 mg/kg q12h of 

gentamicin, for 5 days. At the end of the terapy five treated udder 

quarters had developed both, clinical remission of symptons and 

bacteriological cure at 8, 12, 16, 17 and 21 days, respectively. In six 

udder quarters had no response to gentamicin therapy and, after another 

therapeutic tentative was decided by chemical cauterization with 10% 

iodine solution. Three udder quarters developed necrosis; overt 

ischemia was observed 3-5 days after the onset of therapy. It was 

observed complete sloughing of the affected mammary tissue. 

Additionally to antibiotic therapy, were provided support fluid therapy 

for these cows; one cow died. The results showed that the treatment of 

acute mastitis induced by Sthaphylococcus aureus in early postpartum 

period had no good efficacy, probably due to biological factors of S. 

aureus, in particular in lactant mammary gland and, cow physiological 

conditions at postpartum. Therefore, it is important to provide special 

hygiene environmental conditions for periparturient cows. 

Financial Support: FAPESP 

Key words: cow, acute mastitis, Staphylococcus aureus 

614 Comparison between Delaval Cell Counter DCC and 

Fossomatic 5000 for the Determination of Somatic Cell Count 

of Bovine Milk 

JP. Roy, L. DesCôteaux, D. DuTremblay 

Université de Montréal, Department of clinical science, St-Hyacinthe, 

Canada 

Objective: To compare somatic cell count (SCC) measured by the 

DeLaval cell counter (DCC) and the Fossomatic 5000 (Foss 5000) on 

individual quarter of Holstein cows. 

Materials and methods: This study was part of another study to 

evaluate extended lactational therapy for cows chronically infected 

by S. aureus. Quarter milk samples were collected from 100 cows 

coming from 14 dairy herds in St-Hyacinthe (Québec, Canada) at 3 

sampling periods: 14 days before the beginning of the treatment, 10 and 

31 days after the end of the treatment. At sampling, Bromopol was 

added to milk samples as conservative agent. Milk samples were 

analyzed with the DCC before being submitted within 2 days to 

VALACTA (provincial DHI system) for an evaluation of the SCC by 

the Foss 5000. Coefficients of determination (R2) were calculated 

between DCC and Foss 5000 using real and logarithmic fits of the SCC. 

Results: A total of 827 quarters were sampled and analyzed by both 

methods. The mean values were 2 204 X10 3 c/mL (1 – 29 633 X10 3 

c/mL) and 1 177 X10 3 c/mL (1 - 7 632 X10 3 c/mL) for Foss 5000 and 

DCC, respectively. The median values were 395 X10 3 c/mL and 337 

X10 3 c/mL for Foss 5000 and DCC, respectively. A total of 315 

samples were below 200 X10 3 c/mL and 257 were over 1 000 X10 3 

c/mL using Foss 5000. A total of 331 samples were below 200 X10 3 

c/mL and 230 were over 1 000 X10 3 c/mL for DCC. When all results 

were included in the analysis, coefficients of determination (R2) 

were 0.62 and 0.99 for real and logarithmic fits of the SCC, 

respectively (P < 0.001). Difference in R2 between both fits is 

largely explained by the fact that the maximum value reported by the 

DCC is limited to 4 000 X10 3 c/mL. For somatic cell counts below 1 

000 X10 3 c/mL, the regression coefficient was 0.92 without 

logarithmic conversion. Below 1 000 X10 3 c/mL the DCC gives 

results 10 % lower than the Foss 5000 reader (parameter estimate: 

1.103 ± 0.014) and the standard deviation of the residuals is 108 

X10 3 c/mL. 

Conclusion: DCC is an accurate tool to measure the somatic cell 

counts in quarter milk samples. However, there is limitation for very 

high values and a 10 % correction factor should be applied to the value 

obtained by the DCC. 

The study was supported by DeLaval and Fort Dodge Animal Health 

615 Effect of Systemic Antibacterial Administration during 

Prepartum Period on Coagulase Negative Staphylococcal 

Intramammary Infection in Holstein Heifers 

O. Ataie 1 , P. Hovareshti 2 , M. Bolourchi 2 , A. Barin 2 , A. Gerami 3 , 

A. Niasari-Naslaji 2 

1 

Faculty of Veterinary Medicine, Islamic Azad University, Karaj 

Branch., Clinical Sciences, Karaj, Iran 

2 

Faculty of Veterinary Medicine, University of Tehran, Clinical 

Sciences, Tehran, Iran 

3 

School of Mathematics, University of Tehran, Statistics and 

Computer Science, Tehran, Iran 

Pre- and peripartum intramammary infections may impair mammary 

development and affect milk production after calving in dairy heifers. 

In the present study, The efficacy and cure rate of prepartum 

intramuscular injection of Tylosin and Cefquinome on coagulase 

negative staphylococcal intramammary infection and reducing somatic 

cell counts was evaluated during the early lactation period. A total of 

183 coagulase negative staphylococcal infected quarters from 229 

pregnant heifers were randomly allotted into 3 experimental groups: 

infected quarters that did not receive any treatment (positive control 

group; n=60); infected quarters that received Tylosin (Tylosin group; 

n=61; 10 mg/kg of 20% solution; sid x 3; IM; Tyloject ® ; Razak, Iran) 

or Cefquinome (Cefquinome group; n=62; 1 mg/kg of 2.5% 

suspention; sid x 2; IM; Cobactan ® ; Intervet, The Netherlands). 

Uninfected quarters that did not receive any treatment were considered 

as negative control group (n=64). Treated heifers received Tylosin or 

Cefquinome 10 to 14 days before expected calving date. Antibiotics 

residues were monitored on Day 3 after calving using a commercial test 

(Copan test; Hansen Company, Denmark). Cure rates, based on 

sampling obtained 3 days after calving, were 85.3% and 69.4% in 

Tylosin and Cefquinome treated groups, respectively. At the same 

time, self limited rate in positive control group was 53.3%. Tylosin 

treated heifers had significantly lower somatic cell count scores 

(4.2±0.2) compared to positive control group (6.0±0.21; P

an important part of the pathogens involved in subclinical mastitis. A 

significant number of Klebsiella spp. was also found in samples from 

clinical as well as subclinical infections. 

617 Antibiotic Susceptibility of Staphylococcus aureus Strains 

Isolated from Bovine Mastitis Milk in France 

O. Bidaud 1 , J. Swinkels 2 , M. Bonnier 3 

1 

Intervet Schering-Plough, Farm and Production Animals Business 

Unit, Angers, France 

2 


Unit, Boxmeer, the Netherlands 

3 

Laboratoire D’Analyses d’Ille-et-Vilaine, Microbiology, Rennes, 

France 

Objectives: Despite the current dominance of environmental 

pathogens in clinical mastitis in France, contagious pathogens such as 

Staphylococcus aureus remain important pathogens in both clinical and 

subclinical mastitis. Furthermore, Staphylococcus aureus infections are 

often chronic and difficult to cure. This study determines susceptibility 

of Staphylococcus aureus to antibiotics commonly used for mastitis 

treatment in France. 

Materials and Methods: A total of 94 Staphylococcus aureus isolates 

from samples collected from naturally occurring subclinical and 

clinical mastitis cases in France were cultured and tested for Minimal 

Inhibitory Concentrations (MIC) at Laboratoire D’Analyses d’Ille-et- 

Vilaine in Rennes (LDA 35), from January 2006 to November 2007. 

Antibiotic susceptibility was assessed in vitro for 10 antibiotics 

commonly used in French veterinary practice, including 8 ‚-lactams 

(penicillin G, amoxicillin/clavulanic acid, cloxacillin, nafcillin, 

cephalexin, cefazolin, cephapirin, cefquinome, gentamicin and 

enrofloxacin). The agar dilution method was used to determine MIC 

values. Reference strains (ATCC 29213 and ATCC 25923) were tested 

and the results were compared to CLSI and CA-SFM standards. 

Results: The MIC of all antibiotics tested ranged from 0.0312 to 16 

µg/mL and MIC 90 ranged from 0.125 to 4 µg/mL. In this study, 

cephapirin and enrofloxacin had the lowest MIC 90 (0.125 µg/mL) and 

therefore were the most potent antibiotics against Staphylococcus 

aureus. However, enrofloxacin was also the only antibiotic that 

demonstrated a bimodal distribution indicating the existence of resistant 

strains. For penicillin G, amoxicillin/clavulanic acid, cephalexin, 

cefazolin and enrofloxacin, some strains showed MIC > 2 µg/mL. 

Conclusions: This study shows recent data on susceptibility of 

Staphylococcus aureus to antibiotics frequently used in European 

veterinary practice against mastitis. It confirms the potential efficacy of 

several cephalosporins like cephapirin or cefquinome, and of other 

molecules like nafcillin or gentamicin against Staphylococcus aureus. 

However, some Staphylococcus aureus strains with a high MIC for 

enrofloxacin were found. 

618 Antibiotic Susceptibility of Coagulase Negative Staphylococci 

(CNS) Isolated from Bovine Mastitis Milk in France 

O. Bidaud 1 , J. Swinkels 2 , M. Bonnier 3 

1 



2 

Intervet Schering-Plough, Int. Marketing, Boxmeer, The Netherlands 

3 

Laboratoire D’Analyses d’Ille-et-Vilaine, Microbiology, Rennes, 

France 

Objectives: CNS have been reported as emerging mastitis pathogens. 

The objective was to determine the prevalence of the different CNS 

strains in France and their susceptibility to antibiotics commonly used 

for mastitis treatment in France. 

Materials and Methods: A total of 94 CNS, isolated from milk 

samples taken from mastitis cases in France, were cultured at LDA 35, 

from November 2005 to November 2007. If a CNS was isolated, the 

exact strain was identified using an API ID 32 Staph test. In vitro 

susceptibility was established for 10 antibiotics: penicillin G, 

amoxicillin/clavulanic acid, cloxacillin, nafcillin, cephalexin, 

cefazolin, cephapirin, cefquinome, gentamicin and enrofloxacin. The 

agar dilution method was used to determine MIC. Reference strains 

were tested and the results were compared to CLSI and CA-SFM 

standards. 

Results: The most prevalent isolated CNS strains were S. 

chromogenes (n=25), S. warneri (n=13), S. xylosus (n=11), S. 

epidermidis (n=9), S. simulans (n=8) and some 29 others from 10 


different species.Overall susceptibility of the isolated strains to ‚lactams 

was good, although cephalexin had higher MIC than the 

other cephalosporins tested. Regarding the MIC distribution, 

gentamicin and cephapirin were the most potent antibiotics against 

CNS (MIC 90=0.25 µg/ml). Penicillin G and cloxacillin displayed a 

bimodal distribution indicating the existence of some resistant 

strains. Regarding the 39 highest MIC values (representing the 

strains with highest MIC for each antibiotic), S. epidermidis (that 

represented only 9 strains among the 94 strains tested), was 

responsible for 23 (i.e. 59%) of these highest MIC values. 

Conclusions: This study provides original new French data on the 

antibiotic susceptibility of CNS for antibiotics commonly used in 

European veterinarian practice for the treatment of mastitis. It 

confirms the potential efficacy of several cephalosporins like 

cephapirin, cefquinome and cefazolin, as well as nafcillin, 

enrofloxacin and gentamicin against CNS. This study is also in line 

with previous studies and confirms that MIC of most antibiotics for 

S. epidermidis is higher when compared to the MIC for other CNS 

species. 

619 Uterine Bacteriology Data in Cows with Post Partum Acute 

Metritis Treated with Cobactan ® IU 

B. Huyghe 1 , C. Sa, de 1 , J. Szabo 2 , J. Foldi 3 , P. Cox 1 , J. Swinkels 4 

1 

Intervet Schering-Plough, R&D, Angers, France 

2 

Faculty of Medical Science, Department of Microbiology, Debrecen, 

Hungary 

3 


Unit, Budapest, Hungary 

4 

Intervet Schering-Plough, Int. Marketing, Boxmeer, the Netherlands 

Objectives: A field trial was conducted in 2006 in Hungary 

evaluating the efficacy of Cobactan ® IU in the treatment of acute 

metritis in cows. Results of bacteriological identification of uterine 

pathogens isolated during this trial and their semi-quantitative 

evaluation are presented here. 

Materials and Methods: Cows included in the study were diagnosed 

at 4-14d p.p. with acute metritis, defined as presence of red-brown 

watery foul smelling uterine discharge. At inclusion (D0) 25% of cows 

were randomly selected for intra-uterine swabbing for bacteriological 

investigation (n=33). Thereafter, cows were treated with intra-uterine 

application of 900mg cefquinome (Cobactan ® IU, Intervet). On D21 

post treatment a second swabbing was performed (n=25). The swabs 

were within 24h transported to the diagnostic lab in Portagerm tubes 

(Biomerieux, France) and cultured for identification and semiquantitative 

evaluation. 

Results: At D0, 100% of samples were found to contain various 

bacteria species. On average each swab contained 3.4+0.80 different 

bacteria species, 10 3 -10 5 CFU per swab. E. coli, Prevotella spp. and A. 

pyogenes were found with 75.8%, 81.8% and 33.3% prevalence 

respectively. Fusobacterium spp. was detected in 12.1% of swabs. 

Streptococcus spp. was present in 81.8% of samples, but considered an 

opportunistic rather than metritis pathogen. On D21, the average 

number of different species per swab was reduced to 1.2 +1.20. E. coli 

was no longer detected in 66.7%, A. pyogenes in 100% and Prevotella 

spp. in 53.6% of cases. If bacteria species were still detected, in most 

cases the bacterial load was reduced (at least 3 log CFU decrease per 

swab vs. D0). These results corresponded well with the clinical cure of 

78% observed. 

Conclusions: On D21 after treatment with Cobactan ® IU, 

bacteriological cure or at least 3 log reduction of the bacterial load was 

reached in 63.2%, 88.9% and 100% for Prevotella spp., E. coli and A. 

pyogenes, respectively. 

620 Effects of Subclinical Mastitis during Early Lactation on 

Reproductive Parameters in a Dairy Herd in Iran 

R. Asadpour, M. Tooloei 

Faculty of Veterinary Medicine, University of Tabriz, Clinical 

Sciences, Tabriz, Iran 

Bovine subclinical mastitis is an inflammation of the mammary 

caused by bacteria which occur frequently during early lactation. 

The objective of this study was to determine the effect of subclinical 

mastitis in early lactation on reproductive parameters of dairy 

Holstein cattle. Composite foremilk samples were collected from 60 

Holstein cattle during early lactation between 5 th to 14 th postpartum

days in a dairy farm in Iran from May to December 2007. The 

animals were apparently normal and their milks had no alteration in 

color or other morphologic features. Reproductive parameters of the 

studied animals including services per conception, days open, and 

days to first service were recorded from their hygienic files. The 

milk samples were examined by microbiological analysis. Briefly, 

foremilk sample (10µl) of each cattle plated onto blood agar culture 

medium supplemented with 5% defibrinated sheep blood. Plates 

were incubated at 37 0C and bacterial growth was recorded at 24-h 

intervals for 2d. Bacteria on primary culture medium were identified 

tentatively according to colony morphologic features, hemolytic 

characteristics, Gram stain reaction and catalase test. Gram negative 

isolates were plated on MacConkeys agar. Presence of any 

pathogens in at least two consecutive samples on primary cultures 

was defined as subclinical mastitis. The results were statistically 

analyzed by the SPSS software Ver.11.5. 60% of the samples (36/60) 

had bacterial growth in the primary culture medium. The most 

common mastitis pathogens isolated from samples with subclinical 

mastitis were Staphylococcus aureus and Streptococcus agalactiae. 

Mean Days open of the cattle infected with subclinical mastitis 

(124±6.5 d) were significantly greater compared with uninfected 

animals (98±3 d) (0.01

4 Universidade de Sao Paulo, Clinica Medica, Sao Paulo, Brazil 

The objective of the present study was to determine the quantity of 

somatic cells found in the colostrum of healthy Holstein cows. A 

total of 171 colostrum samples were collected before the first and 

second milking of the cows. Exclusion criteria were physical 

examination, including examination of the udders, and 

bacteriological examination of colostrum. Samples were collected 

and divided in two aliquots. The first one was used in quantitative 

cytology, and was obtained by adding 1mL of colostrum to 1mL of 

phosphate buffered saline (PBS), in glass tubes. The second aliquot, 

used in qualitative cytology, was obtained adding 1mL of colostrum 

to 49 mL of PBS in sterile Falcon tubes. Total leukocyte counts were 

determined by means of direct microscopy, using Rosenfeld stain, 

and differentiating mononuclear and polymorphonuclear leukocytes. 

For qualitative cytology, collostrum smears were produced by 

cytocentrifugation and were then stained by Rosenfeld. Results were 

expressed in relative values (%) for the different types of leukocytes. 

Medians of mononuclear and polymorphonuclear leukocytes per mL 

of colostrum before the first milkiing were equal to 657,064 and 

178,550, and before the second milking, 828,472 and 392,810, 

respectively. There was a statistical difference (p

Staphylococcus spp. (P

incubated for one hour at 37 oC. After this period, samples were 

submitted to a post-incubation viability evaluation through Trypan 

Blue exclusion test. Mean pre-incubation viability rate (± standard 

deviation) of milk cells was 50.17% (±6.50). Mean post-incubation 

viability rates (± standard deviation) of control group (devoid of any 

drug), Drug 1, Drug 2, Drug 3, and Drug 4 were, respectively, 

31.36% (± 3.61); 26.23% (± 4.49); 7.82% (± 2.33); 3.10% (± 0.94); 

and 30.33% (± 4.14). Results show that all treatments decreased 

milk leukocytes viability rates (p

paratuberculosis programmes, and to provide an update on the 

results of the BMQAP. 

Key words: paratuberculosis, certification, surveillance, milk 

quality 

14 Mycobacterium Avium Paratuberculosis Invades the Human 

Gut Epithelium and Elicits Local Inflammatory Response - 

Implications for the Pathogenesis of Crohn’s Disease 

N. Shpigel 1 , L. Golan 1 , A. Livneh 1 , I. Rosenshine 2 

1 

The Hebrew University of Jerusalem, Koret School of Veterinary 

Medicine, Rehovot, Israel 

1 

The Hebrew University of Jerusalem, Faculty of Medicine, 

Jerusalem, Israel 

Crohn’s disease (CD) is a chronic debilitating inflammatory bowel disease 

(IBD) of unknown etiology whose incidence is on the rise worldwide. 

Mycobacterium avium paratuberculosis (MAP) is the etiology of 

inflammatory bowel disease of ruminants, which bears similarity to human 

Crohn’s disease (CD). The role of MAP in CD is highly controversial, it 

was found in the gut and blood of CD patients, but causality was not 

established. To this end we tested the hypothesis that MAP can colonize 

the normal human gut and elicit inflammation and tissue damage. Fully 

developed, germ-free human small intestine and colon were established by 

subcutaneous transplantation of human fetal gut into SCID mice. A total of 

83 intestinal xenografts originating from 9 different fetal donors were 

allowed to develop for 3-4 months and thereafter were infected by direct 

intraluminal inoculation of a bovine strain of MAP. Using 

immunohistochemical methods we have found that in all human donors 

MAP actively invaded the human gut epithelium, with specific tropism to 

the goblet cells of the small intestine. In 3 out of 9 human donors, invading 

MAP induced severe tissue damage and inflammation characterised by 

massive recruitment of macrophages and neutrophils into the gut mucosa 

producing high levels of IL1, TNF and IL6. These observations implicate 

that MAP can specifically colonize the normal human small intestine and 

elicits inflammation and severe mucosal damage. Goblet cell infection by 

MAP might play a key role in the pathogenesis of CD. Furthermore, this 

mouse model will enable to analyze MAP virulence mechanisms in the 

human gut and possible involvement of MAP in CD. 

Key words: paratuberculosis, Crohn’s disease, xenografts 

15 Isolation and Characterization of Verocytotoxin-producing 

Escherichia coli O157 from Turkish Cattle 

O. Aslantas 1 , S. Erdogan 1 , Z. Cantekin 1 , I. Gulacti 2 , G. Evrendilek 3 

1 Mustafa Kemal University, Microbiology, Antakya, Turkey 

2 Firat Univesity, Department of Virology, Elazig, Turkey 

3 Mustafa Kemal University, Food hygiene and Quality Control 

Department, Antakya, Turkey 

The objective of this study was to collect rectal swabs from the cattle in 

a slaughterhouse located in Hatay (Turkey) immediately after slaughter 

for the isolation and characterization of verotoxin-producing 

Escherichia coli 0157 in each month during a 1-year period. The rectal 

swab samples were analyzed for the isolation of E. coli 0157 through 

pre-enrichment, immunomagnetic separation and selective plating on 

CT-SMAC agar. E. coli 0157 was isolated from 77 (13.6%) of the 

samples. The presence of E. coli 0157 changed during a 1-year period, 

in that the occurrence of E. coli 0157 was the highest in July and 

November and lowest in February. A total of 66 isolates out of 77 were 

seroytpe 0157:H7 and 11 were serotype 0157:NM. PCR analysis of E. 

coli 0157 virulence genes revealed that all 0157:H7/NM were positive 

for rbf(O157), 74 positive for EhlyA, 72 positive for eaeA, 62 positive 

for vtx2, and 3 positive for both vtx1 and vtx2. It was presented by 

cytotoxicity tests that many of E. coli 0157 isolates showed high 

cytotoxicity on Vero cells. All of the isolates containing EhlyA showed 

enterohaemolysin production. 

Key words: Escherichia coli O157, Turkey, cattle, IMS, PCR 

16 A Seroepidemiological Study of Bovine Leptospirosis in 

Bouyer Ahmad District, Western Iran 

E. Rahimikia 1 , G. Abdollahpour 2 , E. Mostafavi 3 

1 

Faculty of Agriculture, Azad University of Yasooj, Animal Science, 

Yasooj, Iran 

2 


Science, Tehran, Iran 



Leptospirosis is an infectious disease caused by pathogenic members of 

the genus Leptospira. It affects both man and animal worldwide resulting 

in high morbidity and mortality. Infection in domestic animals especially 

castles can lead to economic loss and pose a potential spread to the 

communities. A serological study was conducted in order to investigate 

the present status of Leptospirosis in cattle in Bouyer Ahmad district in 

western Iran during June to October 2006. A total of 236 samples were 

collected randomly from indigenous cattle of different age and sex; and 

were tested for antibodies against 6 different Leptospira interrogans 

serovars (Gripptyphosa, Icterrohamoragia, Canicola, Hardjo, Ballum 

and Pomona) using a microscopic agglutination test (MAT). The results 

of this study showed that 30 (12.7%) of the samples had a positive 

reaction against one or more serovars. The most prevalent Leptospira 

serovar were Gripptyphosa (7.2%), Canicola (1.7%), Icterrohamoragia 

and Hardjo (1.3%). The less prevalent Leptospira serovar (0.4%) was 

Pomona. In this survey association between serum positive samples and 

exposure to rodents or dogs was statistically significant (P < 0.05). This 

prevalence study indicated there is a high prevalence of Leptospirosis in 

this region. Control of this disease consists of implementing biosecurity 

measures, use of antibiotics to clear carrier states and use of effective 

vaccines. 

Key words: seroepidemiology, leptospirosis, cattle, Bouyer Ahmad, 

Iran 

46 Isolation of Clostridium difficile from Veal Calves 

L. Arroyo 1 , A. Van Dreumel 2 , R. Lothrop 4 , H. Staempfli 3 , J. Weese 1 

1 

Ontario Veterinary College, Department of Pathobiology, Guelph, 

Canada 

2 

University of Guelph, Guelph, Canada 

3 

Ontario Veterinary College, Clinical Studies, Guelph, Canada 

4 

Private, Cambridge, Canada 

Clostridium difficile is an important enteropathogen in humans and some 

domestic animals. It has been isolated and its toxin(s) detected in feces of 

diarrheic and non-diarrheic dairy calves, yet its role as a cause of disease 

in veal calves has not been investigated. This study describes the 

pathological findings of suspected C. difficile-associated gastroenteritis in 

veal calves and characterizes C. difficile isolates obtained from veal 

calves. Six calves were submitted for necropsy examination from a veal 

farm. Gastrointestinal contents were screened for pathogens associated 

with calf diarrhea, including enterotoxigenic E. coli, Salmonella spp. C. 

perfringens, viruses and parasites. Samples from 2 calves were cultured 

for C. difficile. Samples were tested for C. difficile toxins A/B and C. 

perfringens enterotoxin using an ELISA and C. difficile toxins were 

detected in all cases. No other pathogens were identified. Gross lesions 

were similar in all cases and consisted of fibrinous enteritis, colonic 

edema, hemorrhagic enterocolitis, dehydration, and pulmonary 

congestion and edema. Histologically, there were focal areas of mucosal 

erosion and fibrino-cellular exudates, with colonies of clostridia-like 

bacilli in the lumen and on the mucosal surfaces of the small intestine and 

abomasum. There was transmural edema and focal areas of hemorrhage in 

the lamina propria, with congested and thrombosed capillaries. Fecal 

samples were collected at the farm from 24 diarrheic calves for C. difficile 

culture. Three historic isolates recovered from diarrheic calves 4 years 

earlier from the farm were also analyzed. PCR-ribotyping and screening 

for genes encoding toxins A (tcdA), B (tcdB) and binary toxin (cdtB) were 

performed. Clostridium difficile was isolated from the 2 initial calves and 

22/24 (92%) diarrheic calves. Five toxigenic ribotypes were identified 

from the 27 isolates. Sixteen (57%) possessed genes tcdA and tcdB, while 

9 (32%) only possessed genes encoding tcdB and 2 (7.1%) possessed tcdB 

and cdtB genes. Overall, genes encoding tcdA, tcdB and cdtB were present 

in 16 (59%), 27 (100%) and 2 (7%) strains, respectively. Two ribotypes 

accounted for 78% of isolates. This report supports the potential capacity 

of C. difficile to colonize and cause disease in several animal species. 

Further studies of diarrheic and normal veal calves are required to 

elucidate the role of this pathogen as a cause of gastroenteritis and 

diarrhea in veal calves. 

47 Severe Outbreaks of Botulism in Cattle Herds in Flanders : 4 

Case Reports 

M. Goderis, M. Hostens, G. Opsomer 

Ghent University, Dep Reproduction, Obstetrics and Herd Health, 

Merelbeke, Belgium 

Infectious and Zoonotic Deseases (Public Health) 75

In Belgium, an increasing amount of catastrophic botulism outbreaks in 

cattle herds have recently been reported to the Belgian Federal Agency for 

the Safety of the Food Chain (FASFC). The latter confirms that at least in 

Belgium botulism is becoming an emerging disease in cattle resulting in 

large economical loss for the farmers. This presentation describes 4 major 

outbreaks in the Flemish part of Belgium occurring between July 2005 

and February 2007. The average mortality on all farms varied from 27 to 

96 %. The first case involved a mixed dairy-poultry farm that encountered 

a morbidity of 33 % and mortality of 27 %. Two possible causes of 

intoxication have been proposed. One being a severe rainfall spreading 

the toxins from the poultry litter to the dairy stable and maize silage, the 

other is an infected grass silage after fertilization with poultry litter. C. 

botulinum intoxication was diagnosed by identification of C. botulinum 

type C and D toxins in the faeces and the liver of the affected animals and 

in samples of the maize silage. The second dairy farm reported a 

morbidity and mortality of respectively 51 % and 49 %. Here, the 

diagnosis was confirmed by identification of the C. botulinum type D 

toxin in liver and mixed-forage samples. A chicken cock housing a nearby 

water well that provided the drinking water for the cows was found 

positive for the same type of toxin, and was hence seen as the main 

contributor to the catastrophe. On the third farm, a mixed swine-dairy 

herd, the highest morbidity and mortality was seen, being respectively 100 

% and 96 %. The farm had one ensilaged grass pack in which possibly a 

wild animal was encaptured while moving. A sample of this grass silage 

was positive for the C. botulinum type D toxin. The fourth poultry-dairy 

mixed farm previously piled up poultry litter on the place where actually 

the wet brewer grains fed to the dairy cows were stored. On a total of 85 

cows, 67 died leading to a mortality of 79 %. Neurotoxin type D was 

isolated in a wet brewer grain sample leading to the C. botulinum 

diagnosis. The presented outbreaks here encouraged Belgian farmers, 

veterinarians, the Flemish Animal Health Service (DGZ-Vlaanderen) and 

the FASFC on describing risk factors for botulism more profoundly and 

elaborated a strategy leading to a quicker diagnosis and a better approach 

of a C. botulinum outbreak in cattle. These will be discussed in detail. 

Key words: Clostridium botulinum outbreak, cattle, poultry 

48 Molecular Mechanism of Action of Pentosan Polysulfate on the 

Cellular Prion Protein 

V. Stadnyk, C. Mayor,V. Vlizlo 

Institute of Animal Biology UAAS, Scientific Center for Prion 


Objectives of study: It was shown by our previous researches, that 

pentosan polysulfat has the ability to inhibit the expression of cellular 

prion protein (PrP C ) in the prion-replicative organs of laboratory animals, 

that it can be used for the prophylaxis of prion infections, in particular 

BSE. But the molecular mechanism of action of this preparation on PrP C 

until now is not clear. The aim of our work was to find out the molecular 

mechanisms of pentosan polysulfate influencing on PrP C . 

Materials and Methods: for the research of co-operation of pentosan 

polysulfate and PrP C were used the ELISA method, mapping of amino 

acid residues of PrP C , which co-operate with pentosan polisulfate were 

carried out by bioinformatical methods and UV-spectroscopy. 

Results: It is set that PrP C and pentosan polysulfate specifically cooperate 

in vitro. Except this, it is shown that for this co-operation the 

sulfate groups of pentosan are responsible and aminogroups of Arg148 

and Arg151. 

Conclusions: it is set that inhibiting influence of pentosan polysulfate 

on PrP C is predefined, probably, by the direct co-operation of these 

molecules. 

Key words: prion protein, pentosan polysulphate 

49 Distribution of Abnormal Prion Protein in the Brain of Goats 

E. Bouzalas 1 , C. Dovas 1 , S. Kritas 1 , M. Papanastasopoulou 1 , 

D. Papakostaki 2 , E. Chatzinasiou 1 , G. Koptopoulos 1 

1 

Aristotle University of Thessaloniki, School of Veterinary Medicine, 

Department of Microbiology and Infectious Diseases, Thessaloniki, 

Greece 

2 

Veterinary Institute of Infectious and Parasitic Diseases, 


The objective of this study was to assess the distribution of PrP Sc in the 

brain of goats raised in a flock with high incidence of scrapie. Scrapie 

is a progressive fatal neurodegenerative disease affecting both sheep 

and goats. Its diagnosis is mainly based on the detection of the 


abnormal prion protein (PrP Sc ) in the medulla oblongata at the level of 

the obex. The study was undertaken in a semi-extensive flock of goats 

comprised of 250 animals, 105 of which were older than 12 months. All 

animals were euthanized and samples from medulla oblongata were 

taken from 86 out of the 105 goats. Additional samples were collected 

from pons, cerebellum, midbrain at the level of superior colliculus, 

diencephalon at the level of optic chiasm and frontal lobe of cerebrum 

to study the distribution of PrP Sc in these animals. The samples were 

examined by a rapid ELISA (Enzyme-Linked-Immunosorbent-Assay) 

test (EC No 999/2001) and an immunochromatography assay and 

positive samples were additionally confirmed with Western blot (WB) 

and/or immunohistochemistry (IHC). Examination of the obex with the 

rapid ELISA test revealed the presence of PrP Sc in 12 goats (14%). Five 

of them had shown also clinical signs such as disorientation, ataxia, 

apathy and sometimes aggressiveness. In 10 out of those 12 animals, 

PrP Sc was detected by both methods in other parts of the brain besides 

obex. Interestingly, in 3 goats other than the obex positive ones, PrP Sc 

was found in other parts of the brain and mainly in the hypothalamus 

(diencephalon). It was concluded that these findings might question the 

use of obex as the exclusive sampling site for detecting the presence of 

PrP Sc in goats. 

Key words: scrapie, PrP Sc , distribution, brain, goats 

50 Dynamics and Distribution of Prion Infectivity in Sheep Blood 

C. Lacroux 1 , N. Morel 2 , S. Simon 2 , H. Cassard 1 , F. Corbiere 1 , 

J. Mathey 1 , F. Schelcher 1 , F. Lantier 3 , J. Grassi 2 , O. Andréoletti 1 

1 

INRA ENVT UMR 1225, Interactions hôtes-agents pathogénes, 

Toulouse, France 

2 

CEA, SPI, Saclay, France 

3 

INRA, IASP, tours, France 

In sheep prion transmission has been observed by transfusion of whole 

blood sampled at the end of the first half of the incubation phase. 

However, in this species, dynamics and distribution of the infectivity 

presence in blood remained poorly documented. We first characterized 

infectivity dynamics and distribution in blood from sheep experimentally 

challenged by the oral route with a “rapid” strain. In this model clinical 

onset is observed as soon as 160 days post challenge. Inoculated animals 

were sampled every month and blood was fractionated into buffy-coat, 

Ficoll cells and plasma. Moreover T CD4 and CD8 lymphocytes, B 

lymphocytes (CD45r), Mono-nucleated phagocytes (CD14) and Negative 

fraction (containing CD4-, CD8-, CD45r- and CD14- cells) were sorted 

from Ficoll preparations. Each fraction was immunoprecipitated before 

concentration under small volume and IC inoculation to Tg338 mice 

(transgenic ovine PrP gene). Bioassay revealed that, infectivity (i) can be 

detected in this model as early as D60 post challenge (ii) is mainly linked 

to CD14 positive cells. Moreover this experiment provided elements 

indicating that infectivity presence in blood is correlated to PrPsc presence 

in secondary lymphoid tissue. In a second experiment, we investigated 

sheep naturally infected with scrapie (Langlade model). In this model, 

blood transfusion allowed to transmit disease as early as 3 months of age 

and infectivity presence in blood persisted all along the incubation phase. 

Moreover we demonstrated that animals contracting scrapie through 

transfusion were contaminant (through blood) 3 months post transfusion. 

In this model first results of bioassay in Tg338 mice confirmed that most 

infectious blood fractions are the mononucleated cells and more 

particularly CD14. Finally, the efficiency of contamination by blood 

transfusion was established by comparison to IV inoculation using a 

titrated (in Tg338 mice) inoculum. Results revealed that a 200 ml whole 

blood transfusion was as efficient as an IV inoculation with 510 5 DL50, in 

contrast with the low infectivity level observed by direct blood bioassay in 

mice. This element could suggest that infectivity in whole blood is 

supported by a highly efficient mechanism. Both dynamics and nature of 

the infectivity positive fractions are of interest with regards to (i) the 

development and evaluation of diagnostic tests on live animals and (ii) 

human blood sanitary policy. 

Key words : prion, scrapie, sheep, blood, infectivity 

51 Retrospective Study of BSE Surveillance in Canada from 1992 

to 2005 

J. Kellar 1 , J. Paré 2 , L. Myers 3 

1 

Canadian Food Inspection Agency, Animal Products Directorate, 

Ottawa, Canada 

2 

Canadian Food Inspection Agency, Epidemiology and Surveillance 

Section, Saint-Hyacinthe, Canada

3 Canadian Food Inspection Agency, Epidemiology and Surveillance 

Section, Ottawa, Canada 

Bovine Spongiform Encephalopathy (BSE) was first diagnosed in 

Canada in 1993, in a cow imported from the United Kingdom. In 2003, 

the first indigenous case of BSE was diagnosed. To the end of 2007, a 

total of 12 indigenous cases of BSE had been diagnosed in cattle born 

in Western Canada. From the time Canada made BSE reportable in 

1990, surveillance has evolved from passive to active, with a markedly 

greater intensity following the discovery of the first indigenous case. 

To assess Canada’s true prevalence of BSE and the effectiveness of its 

surveillance, a retrospective study was designed based on the random 

selection of 3,712 samples from the 95,383 cattle brain samples 

submitted for BSE testing between 1992 and 2005. The sampling plan 

was based on an a priori estimation of the proportional distribution of 

National, Western and Eastern BSE surveillance samples by Office 

International des Epizooties (OIE) surveillance stream (clinical 

suspect, casualty slaughter, fallen stock, and routine slaughter). For 

each sample, data was obtained by record tracing and interviewing of 

identifiable current and original owners. Information recorded 

comprised sampling site and date, age of the animal, date of birth, 

breed, production type, province of sampling, province of birth, clinical 

signs preceding and reason for death, diagnostic test used and test 

result. Each sample was classified by OIE surveillance stream. 

Analyses on a National, Western and Eastern basis highlighted regional 

variability among surveillance strategies. Reflecting underlying 

population demographics and market flows, Eastern Canadian 

surveillance emphasized deadstock/renderer sampling (70.5%) while 

surveillance in Western Canada focused on on-farm sampling (84.1%). 

Sampling in Western Canada was predominantly from beef cattle 

(78.3%), with a mean age at sampling of 8.1 years. In Eastern Canada, 

the majority of samples originated from dairy cattle (84%), with a mean 

age at sampling of 4.7 years. The majority of samples examined were 

classified as casualty slaughter (63.7% in Eastern Canada and 80.4% in 

Western Canada). National, Eastern Canada and Western Canada 

prevalence estimates were determined. Results of the study have 

contributed to a broader assessment of the natural history of BSE in 

North America. 

86 Pregsure Trivalent Induces Much Higher Neutralising 

Antibody Titres against BVDV-2 Strains than Monovalent 

BVD Vaccines 

R. Raue, D. Hurst, J. Salt 

Pfizer Animal Health, R&D, Sandwich, United Kingdom 

Objectives: Bovine viral diarrhoea virus (BVDV) is one of the most 

common and economically important viruses causing infections in 

cattle. Recently, severe outbreaks of BVDV-2 have been described in 

the UK. In addition, isolation of BVDV-2 has been reported in most 

European countries. As vaccination against BVDV is most often 

performed with inactivated vaccines, neutralising antibodies can be a 

good in vitro indicator of the ability to protect against a certain BVDV 

strain or type. This study was designed to compare the neutralisation 

antibody titres induced after vaccination with a novel vaccine PregSure 

Trivalent (Pfizer Animal Health), Bovilis BVD (Intervet) and Bovidec 

(Novartis Animal Health) against a panel of BVDV-1 and 2 strains. 

Materials and Method: Serum samples were collected from calves 3 

weeks after vaccination either with PregSure Trivalent, Bovilis BVD or 

Bovidec according to the recommendations of the manufacturer. They 

were tested in a serum neutralisation test against 11 BVDV-1 strains, 

representing genotypes 1a, 1b and 1d to 1i, as well as 4 BVDV-2 strains 

from Europe and the US. 

Results: The geometric mean titre in log2 for the 11 BVDV-1 strains 

were between 7.3 and 10.8 for PregSure Trivalent, 5.7 and 9.5 for 

Bovilis BVD and 4.0 and 8.2 for Bovidec. With all vaccine sera 

BVDV-1h strain G-AU was neutralised best and BVDV-1d strain 

PI810 worst. The geometric mean titre in log2 for the 4 BVDV-2 

strains where between 8.5 and 10.1 for PregSure Trivalent, 1.9 and 5.1 

for Bovilis BVD and 1.2 and 2.2 for Bovidec. Therefore, PregSure 

Trivalent serum samples showed an equal ability to neutralise BVDV- 

1 and 2 strains whereas Bovilis BVD and Bovidec serum samples 

showed a reduction in the ability to neutralise BVDV-2 strains by at 

least 4 log2. In some cases Bovilis BVD and Bovidec vaccine sera were 

not able to neutralise BVDV-2 strains at all. 

Conclusions: The results of this study demonstrate that the level of 

neutralising antibodies after vaccination with PregSure Trivalent is 

remarkably higher against BVDV-2 strains than those of monovalent 

inactivated BVD vaccines. In combination with the demonstration of 

12 month protection against BVDV-1 and 2 by challenge, this data 

indicate that the novel vaccine PregSure Trivalent gives a 

comprehensive cover against all types of BVDV field infections. 

Key words: PregSure Trivalent, BVDV-1, BVDV-2, neutralisation 

87 Monitoring of a BVDV Infection in a Vaccinated Herd by 

Testing of Milk for Antibodies against NS3 

B. Makoschey 1 , H. Kuijk 2 , J. Mars 3 , W. bij de Weg 2 , P. Franken 3 

1 

Intervet-Schering Plough, Int. Marketing, Boxmeer, Netherlands 

2 

Intervet-Schering Plough, Intervet Netherlands, Boxmeer, 

Netherlands 

3 

Animal Health Service Deventer, Deventer, Netherlands 

Objectives: It has been demonstrated earlier, that an inactivated 

vaccine against the bovine viral diarrhoea virus (BVDV) in 

combination with a suitable test for antibodies against non structural 

proteins (NS) exhibits properties of a marker vaccine: animals usually 

remain seronegative after vaccination but develop NS specific 

antibodies after field virus infection. In this study, it should be 

established, whether the NS3 antibody testing of individual or bulk 

milk samples is likewise suitable for differentiation between BVDV 

vaccinated and field virus infected animals. 

Materials and Methods: The study was performed on a Dutch dairy 

farm. After being certified as BVDV-virus-free in 2002, the farm 

experienced a new outbreak of BVDV in 2005. A herd vaccination 

program was initiated using an inactivated BVDV vaccine (Bovilis R 

BVD, Intervet International). Seventeen cows were selected for further 

follow up: blood samples, individual milk samples and bulk milk 

samples were collected at pre-set time points. BVDV neutralizing 

antibody titers were measured and the NS3 antibody response was 

measured in a commercial ELISA test (Cedi-testR BVDV, Cedi 

Diagnostics, Lelystad, The Netherlands). 

Results: All animals that were seronegative before vaccination 

developed BVDV neutralizing antibodies after immunization. Three of 

the ten vaccinated animals showed an increase in the titers of BVDV 

neutralising antibodies, between study days 56 and 210, indicating that 

they experienced a field virus infection. All samples from animals that 

were vaccinated but not infected were tested negative for NS3 specific 

antibodies at all time points. The samples from the three animals that 

were infected after the vaccination were tested negative for NS3 

specific antibodies at Day 0, 28 and 56, but were positive thereafter. In 

contrast, all serum samples taken from animals that have been infected 

with BVDV field virus were tested positive for NS3 specific antibodies. 

The overall majority (37/44) of the milk samples for these animals were 

also tested positive in the NS3 ELISA. Likewise, the bulk milk samples 

were positive in the NS3 ELISA, with the exception of the sample 

taken at study day 28. 

Conclusions: The results obtained demonstrate that a field virus 

infection with BVDV can be monitored by measurement of NS 

antibodies in blood or (bulk) milk samples, even in a vaccinated herd. 

88 Transmission of Bovine Viral Diarrhoea Virus (BVDV) from 

Persistently Infected Calves to Naive in-contact Cattle is Very 

Rapid and Efficient 

B. Makoschey 1 , M. Janssen 2 

1 Intervet-Schering-Plough, Int. Marketing, Boxmeer, Netherlands 

2 Intervet-Schering Plough, R&D, Boxmeer, Netherlands 

Objectives: The transmission of bovine viral diarrhoea virus (BVDV) 

from persistently infected (PI) calves to in-contact animals is generally 

considered to happen very efficiently. It should be determined, whether 

naive animals housed in adjacent pens to a PI are readily infected or 

whether housing in the same pen is required for efficient infection. 

Materials and Methods: Four heifers at the age of 10 months or older, 

negative for antibodies against BVDV, and free of BVDV and one three 

months old PI calf were included in the study. The heifers were housed as 

one group in a pen. The PI calf was housed in an adjacent pen. The animals 

were allowed to have direct contact through the fence. Blood samples were 

taken weekly and BVDV neutralising antibodies were determined. 

Results: All four heifers seroconverted within four weeks after 

introduction of the PI calf. These results, indicate that the infection with 

BVDV had occurred during the first week after the first contacts with 

the PI animal. 


Conclusion: It can be concluded that transmission of BVDV from PI 

animals to naive in-contact animals occurred in a very efficacious and 

fast manner, even if the animals are housed in adjacent pens. The 

practical implications of these findings for BVDV control programs 

will be discussed. 

89 BVDV Type 1 and 2 Fetal Protection following the Use of a 

Modified Live Viral Vaccine 

W. Xue, D. Mattick, L. Smith, J. Maxwell 

Intervet-Schering Plough, R&D Desoto, Desoto, United States 

Objectives: Previously, it has been demonstrated that pre-breeding 

vaccination with a modified live virus (MLV) vaccine (Vista R 5 SQ, 

Intervet International) containing BVDV Type 1 and Type 2 strains, 

offers protection against persistently infected fetuses when dams are 

challenged with BVDV type 1 and type 2 in the first trimester. The 

animal trials presented here were designed to demonstrate the efficacy 

of the MLV vaccine in protecting fetuses from infection with BVDV 

type 1 or type 2, when pregnant cows were challenged in the last 

trimester, at 170 days of gestation. 

Materials and Methods: Heifers, seronegative to BVDV, were 

vaccinated at pre-breeding, and then bred naturally 4 weeks postvaccination. 

Eighty-three pregnant heifers were challenged with 

either type 1 (37 in total with 25 vaccinated and 12 controls), or type 

2 (46 in total with 28 vaccinated and 18 controls) non-cytopathic 

(ncp) BVDV field isolates at about 170 days of gestation. Fetuses 

were collected from the challenged heifers 60 days post-challenge 

(for type 2 BVDV challenge), and newborn calves were collected 

prior to taking colostrum (for type 1 BVDV challenge). Serum 

samples were taken from the fetuses or calves for a BVDV serum 

neutralizing (SN) antibody test. Tissue samples of thymus, lung, 

spleen and kidney were collected from the fetuses or calves, and 

were used for virus isolation. 

Results: Results of SN antibody tests revealed that 100% of fetuses 

or calves from control heifers developed SN antibody to BVDV. 

However, only 4 out of 22 calves born to vaccinated heifers were 

positive for SN antibody in the type 1 BVDV challenge study, and 7 

of 28 fetuses from vaccinated heifers were positive for the SN 

antibody in the type 2 BVDV challenge study. Virus isolation from 

calves’ tissue samples demonstrated the presence of type 1 BVDV in 

5 of 12 calves from control heifers and none of the calves from 

vaccinated heifers. The type 2 BVDV was isolated from 17 of 18 

fetuses in the control group, and 2 of 28 fetuses in the vaccinate 

group. 

Conclusions: Results demonstrated that MLV vaccine Vista R 5 SQ 

prevented fetal infection by at least 82% to type 1 BVDV, and 75% to 

type 2 BVDV, when the dams were exposed to BVDV at about 170 

days of gestation. Vaccine Vista R 5 SQ is the first MLV BVDV vaccine 

that has proven to prevent fetal infection in the last trimester of 

pregnancy. 

90 Fetal Protection against Bovine Viral Diarrhea Virus Type 1 

Infection Using One Shot of the Mucosiffa Modified-live Virus 

Vaccine 

G. Meyer 1 , M. Deplanche 2 , D. Roux 1 , M. Moulignie 2 , 

D. Raboisson 1 , N. Hagen-Picard 1 , P. Mathevet 3 , F. Schelcher 1 

1 

National Veterinary School of Toulouse, Pathology of Ruminants, 


2 

National Institure of Agronomical Research, Animal Health 

Department, Toulouse, France 

3 

Merial Animal Health Limited, Lyon, France 

Objective: To evaluate the efficacy of a commercially available live 

Bovine Viral Diarrhea virus (BVDV) vaccine to protect against fetal 

infection in pregnant cattle. 

Method: Thirty beef heifers were allocated into 2 groups. One group 

(18 heifers) was vaccinated against BVDV with one shot of a 

commercially (Mucosiffa R , Merial) live vaccine according 

manufacturer’s recommendations, and the other group (12 heifers) 

served as nonvaccinated control cattle. The vaccine contained 10 3.5 

TCID50 per vaccination dose (2.0 ml) of cp BVDV-1 Oregon C 24 V 

strain. One month later, estrus was induced and the heifers were bred. 

Three cycles of insemination were performed. Pregnancy was 

confirmed by transrectal ultrasonographic examination and bovine 

pregnancy-associated glycoprotein quantification. Four months after 


vaccination, 13 vaccinated and 9 non vaccinated pregnant heifers were 

intranasally challenged with 10 6 TCID 50 of the BVDV-I 22146/Han 

strain, on days 49-96 of gestation. Seroconversion, clinical signs, 

mortality, abortion rate, hematology, viremia, and fetal infection were 

studied. 

Results: By ELISA or neutralization assay, all vaccinated animals 

seroconverted to BVDV four weeks after immunization. 

Neutralizing antibodies reached a plateau two month after 

vaccination with titers ranging from 5970 to 14159 ED50/ml. 

Challenge was followed by a rise of 0.57 log of neutralizing antibody 

titer for vaccinated animals and by seroconversion for control 

heifers. Post-challenge, all heifers were free from clinical signs of 

BVD. Viremia was not detected by real time RT-PCR for 18 days 

post-challenge in any of the vaccinated heifers. In contrast, 100% of 

the control animals were BVDV viremic on at least one day between 

days 4 to 18 post-challenge. Transient leucopenia with lymphopenia 

was observed for heifers of the control group but not of the 

vaccinated one. BVDV type 1 infection led to abortion only in one 

unvaccinated heifer, 47 days after challenge and virus was detected 

by real time RT-PCR in the spleen of aborted fetus. Three months 

after challenge, all fetuses were harvested from heifers and tested for 

evidence of BVDV infection. BVDV was detected with high viral 

charges by qRT-PCR in the spleen, thymus, cerebellum and placenta 

from 8 of 8 fetuses obtained from control heifers but from none of 

the fetuses obtained from the 13 vaccinated heifers. 

Conclusion: The commercial monovalent Mucosiffa R live vaccine is 

safe and fully protective against fetal BVDV type 1 infection and 

abortion. 

Key words: bovine viral diarrhea, vaccination, fetal protection 

91 Impact of Live and Killed Marker IBR Booster Vaccinations 

on BoHV-1 Shedding after Primovaccination with Live 

Marker IBR Vaccine 

M. Pearce, I. Vangeel 

Pfizer Animal Health, Sandwich, United Kingdom 

For BoHV-1 control it is important to minimise virus shedding from 

infected cattle. It has been suggested that killed vaccines are more 

effective at suppressing BoHV-1 shedding than live vaccines. This 

study compared the impact of live and killed marker IBR booster 

vaccines on BoHV-1 shedding after challenge and reactivation 

following primovaccination with live marker IBR vaccine. Twenty 

BVDV free calves aged 12-14 weeks with no antibodies against 

BoHV-1 and BVDV were given 2 doses of live IBR vaccine IM 3 

weeks apart. Six months after primovaccination, 10 calves received 

one IM dose of live IBR vaccine; the other 10 calves received one 

dose of killed IBR vaccine. Live vaccine was Rispoval ® IBR marker 

vivum; killed vaccine was Rispoval ® IBR marker inactivatum. Ten 

control calves received saline IM at the time of each vaccination. Six 

months after booster vaccination all cattle were challenged 

intranasally with 10 8.0 TCID 50 BoHV-1, Iowa strain. Six weeks after 

challenge, calves were given dexamethasone at 0.1 mg/kg 

bodyweight IV daily for five days. Cattle were blood sampled 

weekly for three weeks after booster vaccination and tested for 

neutralising antibodies against BoHV-1. After challenge and 

reactivation, nasopharyngeal swabs were taken daily for 21 days and 

tested for the presence of BoHV-1 by virus isolation. Boosting with 

inactivated vaccine gave higher titres of BoHV-1 neutralising 

antibodies (P

131 Evaluation of the Exhaustion Dynamics of MDA in IBR Cases 

I. Szabó 1 , T. Molnár 2 , T. Abonyi 3 

1 Pfizer NEER Region, Paris, France 

2 MgSzH Diagnosztikai Igazgatóság, Budapest, Hungary 

3 Pfizer Ltd, Budapest, Hungary 

Infectious bovine rhinotracheitis (IBR) causes substantial economic 

losses in cattle herds. The key elements of successful eradication are: 

(a) in IBR-free herds, preventing the introduction of infection and 

maintaining disease-free status, (b) in infected herds, starting an 

eradication programme as soon as possible, determining the spread of 

virus within the herd, applying a management-adapted immunisation 

programme, and breaking the chain of infection in a reliable manner. 

The knowledge of maternal immunity status is essential for accurately 

determining the spread of virus and the age group in which basic 

immunisation should be started. In eradication programmes using 

marker vaccination, it is important to prevent the infection of the 

progeny with virulent virus. The duration of maternally derived 

immunity is extremely important in terms of calf vaccination. The 

primary objective of this work was to collect data on the duration of 

maternal immunity in Hungary, where about 75% of the large dairy 

herds are infected with virulent IBR virus at present. In 17 large cattle 

herds infected with IBR virus, located in Hungary, blood samples were 

taken from 5-10 one- to six-month-old calves per age group. The IBR 

antibody levels of the blood samples were determined by a 

discriminative ELISA. The results clearly show that in 10% of the 

herds the maternal antibodies disappear already by 2 months of age, 

while in 19%, 44% and 73% of the herds the cessation of maternal 

immunity can be observed at 3, 4 and 5 months of age, respectively. On 

two farms the maternally derived protection was present even at 6 

months of age. The findings on some farms indicate that after the 

disappearance of maternal antibodies by 3-4 months of age, in the 

presence of conditions facilitating the spread of virulent virus calves 

may become infected with virulent IBR virus already by 6 months of 

age. These findings are important with a view to the successful In 

conclusion, for the elaboration and successful 

132 Differentiation of Malignant Catarrhal Fever, Mucosal 

Disease and Bluetongue Based on the Retrospective Study of 

115 Cases 

R. Bexiga 1 , H. Guyot 2 , C. Saegerman 2 , A. Mauroy 2 , F. Rollin 2 , 

E. Thiry 2 , A. Philbey 1 , D. Mellor 1 , D. Barrett 1 , K. Ellis 1 

1 University of Glasgow Veterinary Faculty, Animal Production and 

Public Health, Glasgow, United Kingdom 

2 University of Liege, Clinic for ruminants, Liege, Belgium 

The clinical differentiation of bluetongue (BT) in the form currently 

seen in northern Europe is highly relevant, especially from malignant 

catarrhal fever (MCF) and mucosal disease (MD), two endemic diseases 

in most of Europe. The aim of this study was to characterise the clinical 

findings typical of cases of MCF and MD, to compare these to clinical 

findings characteristic of the recent BT outbreak in northern Europe. 

Cases of MD (n=66) and MCF (n=11) were selected from records of 

cattle admitted to the University of Glasgow Veterinary School, from 

January 1989 to June 2007. Selected cases of MD had a postmortem 

diagnosis of MD, were antigen positive and antibody negative for 

bovine viral diarrhoea virus (BVDV) by ELISA. Selected cases of MCF 

had a postmortem diagnosis of MCF, were positive for antibody against 

ovine herpesvirus-type 2 (OHV-2) by an immunofluorescent antibody 

test or for OHV-2 nucleic acid by PCR and were antigen negative for 

BVDV. Cases of BT (n=38) were selected from records of cattle 

examined by the ambulatory clinic of the Faculty of Veterinary 

Medicine of Liege from August 2006 to February 2007. Selected cases 

had a clinical diagnosis of BT and a positive test result either for 

detection of antibody by competitive ELISA or viral nucleic acid by RT- 

PCR. Case data were summarised to determine the most common 

presenting signs for each condition. A Freeman-Halton extension of the 

Fisher exact test was performed to test for significant differences in 

frequency of clinical signs between the three conditions. Clinical signs 

for which there was a significant difference in the proportion of cases 

presenting in each category were appetite, demeanour, size for age, 

faecal appearance, adventitious lung sounds, corneal opacity, nasal 

discharge, halitosis, bilateral lymph node enlargement and interdigital 

ulceration (P

calves recovered after 4 to 10 days, respectively. Repeated 

endoscopic examinations demonstrated that the oesophageal tone 

gradually normalised. Oesophageal paresis in cattle has been 

described as a sequel to oesophageal obstruction or caused by 

metabolic, neurological or neuromuscular disorders. Due to the 

absence of oesophageal obstruction, hypocalcemia or signs of 

encephalitis, Clostridium botulinum type B intoxication was initially 

suspected. In rumen fluid and faeces of one calf, neurotoxins 

belonging to group C/D and a toxigenic C. botulinum strain 

belonging to type A,B or E were found. In rumen fluid and faeces of 

the other calves a substance toxic for mice was isolated, that was not 

neutralisable by botulinum antitoxins. Because of the BTV serotype 8 

epidemic in the region, the calves were tested for BTV by PCR: all 

calves were antigen positive. At necropsy only aspiration pneumonia 

of the cranial lung lobes was found. Histology of the oesophagus 

revealed atrophy and fibrosis of the tunica muscularis. The entire 

oesophagus was affected, but the pathology was most prominent in 

the distal part. In this segment oval calcified structures (80 x 50 µm) 

were found, resembling disintegrated and calcified sarcocysts. No 

vasculitis nor neuronal damage (synaptophysin staining) could be 

demonstrated. Because of the inconclusive results of the C. botulinum 

analysis, the presence of histological changes in the oesophageal 

musculature and the demonstration of BTV infection, a tentative 

diagnosis of oesophageal paresis caused by BTV was formulated. In 

literature this phenomenon has incidentally been reported previously 

and therefore the clinical symptoms of regurgitation caused by 

oesophageal pathology may be included in the list of BTV-induced 

pathology in the future. 

Key words: regurgitation, oesophageal dilatation, blue tongue 

serotype 8, cattle 

135 Seroepidemiology of Bovine Leukemia Virus (BLV) Infection 

in the North Eastern Provinces of Iran 

A. Haghparast 1 , Gh. Mohammadi 2 , Sh. Mousavi 1 

1 Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, 

Department of Pathobiology, Mashhad, Iran 


Department of Clinical Sciences, Mashhad, Iran 

Bovine Leukemia Virus (BLV) is an exogenous c-type oncovirus in the 

retroviridae family that causes a chronic infection in cattle, which 

develops in three possible pathological forms: asymptomatic course, 

persistent lymphocytosis and lymphosarcoma. Once infected, cattle 

remain virus carriers for life and start to show a serological reaction 

within a few weeks after infection. This disease causes significant 

economic losses associated with the costs of control and eradication, 

loss in milk production and difficulties in exports. The Agar Gel 

Immuno Diffusion (AGID) and ELISA tests have been the serological 

choice assays for routine diagnosis of serum samples. However, due to 

the higher sensitivity of ELISA, in recent years this method has 

replaced the AGID in seroepidemiological studies for control and 

eradication programe. Prevalence studies on BLV infection has not 

been conducted in the eastern part of Iran. This study was designed to 

determine the prevalence of BLV in Khorasan Razavi and Khorasan 

Shomali provinces which are the main provinces located in the north 

east of Iran. During summer 2007, 429 blood samples were randomly 

taken from industrial dairy herds of these two provinces and subjected 

to ELISA test (indirect sandwich ELISA). 25.41% of these samples 

showed positive reaction. The positive results in Khorasane Razavi and 

Khorasan Shomali provinces were 29.83% and 1.5% , respectively. 

Moreover, 64.7% of the herds tested, had at least one positive cow. The 

overall true prevalence was 24.63%. Statistical analysis revealed a 

direct correlation between age and occurrence of BLV. Significant 

correlation was also found between herd size and BLV infection. In 

another words, the large herds had the most BLV positive cases. 

Moreover, number of parity was also shown to have a direct correlation 

with BLV seropositivity. Results of Odd ratios analysis which is an 

indication of possible risk factors, confirmed the above findings. Taken 

together, the results presented in this study along with similar studies in 

recent years in Iran confirm an alarming increase in prevalence of BLV 

infection among dairy herds. Therefore, control strategies based on test 

and implementation of corrective management should be taken in order 

to downgrade and possibly eliminate this economically hardship 

disease. 

Key words: BLV, seroepidemiology, Khorasan Razavi, Khorasan 

Shomali, Iran 


136 Individual and Herd Prevalence of Bovine Leukemia Virus 

Infection in Chilean Dairy Herds 

G. Monti, M. Grau 

Universidad Austral de Chile, Institute Preventive Veterinary 

Medicine, Valdivia, Chile 

Objectives of study: The aim of this study was to assess herd and 

within-herd prevalence of Bovine Leukemia Virus (BLV) infection in 

Chilean dairy herds. 

Materials and Methods: Herd and individual level prevalence was 

estimated with a cross-sectional random sample of 80 dairy herds in the 

main dairy area of the country (X° region) performed during 2007. 

Sample was stratified according to herd size in large, medium and small 

and it was considered the proportion of each class in the population of 

the herds of the area (2%, 13% and 85% respectively). All selected 

herds were commercial dairy herds that are registered at the regional 

offices of the Servicio Agrícola y Ganadero (SAG) from the Ministry of 

Agriculture of Chile. SAG also has records of the Milk Collection 

Centers (CALS) where smallholders deliver their milk. To avoid 

interferences with passive antibodies animal older than 6 months will 

be candidates to be tested. A commercial ELISA (IDEXX-BOMMELI) 

for serum samples was used for assess BLV status. The herd and 

individual-level true prevalence were estimated by using a Bayesian 

approach. Briefly, we modelled that n animals were sampled randomly 

from herds. Model estimate prevalence distribution based in the result 

of one test applied to individual animals, assuming that prevalence is a 

mixture of point mass at zero and an continuous Beta distribution on 

(0,1). Independent prior distribution were assumed for test sensitivity 

and specificity and modelled as beta distributions also. 

Results: The results showed a high degree of variation in herd and 

cow-level of BLV prevalence. The posterior median and 95% 

credibility interval for Herd-prevalence was 48% (20.9, 78.4). Also the 

predictive probability that a randomly selected herd in the region had 

prevalence less/equal than 5% is 70% and prevalence less/equal than 

50% is 98%, and it is less that those of neighboring countries. Larger 

and specialized dairy herds showed larger prevalences. 

Conclusions: The cross-sectional study resulted in an estimation of the 

true prevalence estimates of BLV for one of the main dairy regions in 

Chile. The results showed the degree of variation in herd and cow-level 

BLV prevalence suggesting that a compulsory control program is 

feasible and small farmers have good opportunities to add value to their 

production by enrolling a free-herd certification schema. 

Key words: Bovine Leukemia Virus, prevalence, Bayesian approach 

137 Q Fever (Coxiella Burnetii) Eradication in a Dairy Herd by using 

Vaccination with a Phase 1 Vaccine (Coxevac ® , Ceva S.A.) 

Ph. Camuset 1 , D. Remmy 2 

1 Cabinet Vétérinaire, Yvetot, France 

2 CEVA Santé Animale, Libourne, France 

Objective of the study: In a dairy herd followed for fecundity since 

1985, Coxiella burnetii was found in April 2002 and March 2004. That 

analysis was initiated after a deterioration of the fertility results and 

early and late abortions of the dairy cows. In a second step, a protocol 

aiming to eradicate the infection by culling and vaccination was 

performed. 

Material and Methods: The herd is composed of about 30 dairy cows 

producing around 9000 kg of milk per lactation. Since Sept. 2004, the 

dairy cows and the heifers have been vaccinated 2 times/3 weeks of 

interval by sub-cutaneous route with the vaccine Coxevac ® (CEVA S. 

A.), preferentially at W2 and W5 post partum (cows) and before first 

insemination (heifers). A booster injection between W2 and W5 postpartum 

was yearly planned. Post-partum, an individual diagnostic was 

initiated with research of the bacteria by real time PCR in the milk 

mixture of the 4 quarters at D0, D3, W1, W3 and W8 and in the vaginal 

mucus between D0 and D3. 

Results-discussion: During the first year of study, among the 46 cows 

studied (10 first lact.), 15 cows (2 first lact.) excreted at least one time 

the bacteria, 10 several times. The third year, only 3 cows excreted (no 

first lact.); among them, two that have excreted several times have been 

culled. During the study period, only one vaccinated cow, not excreting 

the previous lactation, became a new excreting (only in the vaginal 

mucus); on the other hand, four cows seem healthy again, they had only 

excreted in a previous lactation around calving always in vaginal 

mucus, sometimes in milk. They have never re-excreted later. For

fertility and fecundity, the performances (% of anoestrus, early and late 

metritis and abortions) are better since the use of the vaccination and 

the culling of the excreting cows. Those results must be put in 

perspective according to the sensibility of the Coxiella b. diagnosis 

because of the intermittence of shedding. For the performances of 

reproduction in high production dairy herds, the influence of herd 

management and feeding is able to put in perspective the only influence 

of the eradication of Q fever in that herd. 

Conclusion: Either for human health or for zootechnic and economic 

reasons, the use of a diagnostic method towards Coxiella burnetii 

shedding animals, their vaccination then their progressive elimination 

appear to be a feasible process by using the vaccine Coxevac ® (CEVA 

S. A.). 

Key words: Q fever, bovine, vaccination, Coxiella burnetii 

175 Serological Testing of Primiparous Cows and Young Cattle 

before Pregnancy is Useful to Detect Acute Q-Fever Infection 

within Herds 

J. Boettcher, A. Vossen, A. Gangl, N. Meier, A. Schneppe, 

G. Wittkowski 

Bavarian Animal Health Service, Poing, Germany 

As Q-Fever is a zoonosis knowledge about acute infection within 

cattle herds is of special interest. To estimate prevalence of Q-Fever 

in Bavaria 21858 sera from 765 herds were tested in ELISA (Idexx; 

plates coated with phase I and II-antigen) for antibodies. 

Seroprevalence (SerP%) was 14.5%±0,2% and 62.5%±1,4% of the 

herds were seropositive. Testing 799 cows in autumn 2003 and 2004 

revealed a rate of seroconversion of 4.6% (CI95% 2.9-5.9%) p.a. To 

detect age-related effects 2819 sera were analysed. Decreasing 

SerP% until 6 month of age (mona) is explained by maternally 

derived antibodies. A low level SerP% (

disease. Thus losses were estimated to exceed 2M $US. This is the first 

report of an outbreak of EHDV-7 is recorded in cattle in Israel. 

Key words: EHDV, BTV, spatial, wild ruminanta, wind 

178 Evaluation of a New Respiratory Syncytial Virus Vaccine 

Candidate 

G. Meyer 1 , M. Deplanche 2 , C. Dubuquoy 3 , M. Soulestin 1 , 

G. Durand 3 , N. Castagne 3 , J. Bernard 3 , P. Bernardet 4 , S. Riffault 3 , 

JF. Eleouet 3 

1 



2 

National Institure of Agronomical Research, Pathology of 

Ruminants, Toulouse, France 

3 


Department, Jouy en Josas, France 

4 


Department, Tours, France 

Objective: To evaluate the efficacy of a new vaccine candidate against 

Respiratory Syncytial Virus (RSV) based on N protein nanoparticles. 

Method: The vaccine was constituted by sub-nucleocapsid ring 

structures (N SRS), each ring being composed by 10 or 11 RSV 

nucleoproteins surrounding a bacterial RNA of 70 bases (Patent WO 

2006/117456). Twenty four one-month old calves, free of bovine RSV 

antibodies, were divided in three groups of 8 calves. The first group 

was vaccinated twice (4 weeks interval) with 2 mg of N SRS protein 

with Montanide-ISA70 VG adjuvant by the intramuscular route. The 

second group received at four weeks interval the same vaccine 

formulation and 10 mg of N SRS protein with Montanide-IMS 4112 

VG adjuvant by the intranasal route. The last group served as nonvaccinated 

control calves. One month later, all calves were challenged 

by intranasal and intratracheal routes with 10 5 UFP of the BRSV 3761 

strain. Clinical examination was performed for 21 days post challenge. 

Virus infection was studied by real time PCR on nasal swabs from 3 

days before to 21 days post challenge. Bronchoalveolar lavages and 

lung biopsies were obtained from two calves per group euthanized at 6 

days post challenge and were used for virus quantification. RSV 

antibody response (ELISA) was studied and lymphoproliferation 

studies were performed on maxillary, mediastinal and tracheobronchial 

lymphatic nodes. 

Results: All vaccinated calves developed anti-N antibodies in blood 

and nasal secretions following vaccination. A memory cellular 

response was also detected in the sub-maxillary lymph node of 

vaccinated calves. Clinical examination demonstrated a moderate 

pathology for each group with significant differences on days 5 and 6 

post challenge between the two vaccinated groups and the control one. 

Viral quantification showed lower viral loads in nasal secretions for the 

two vaccinated groups, with significant differences at days 3, 4 and 11 

post challenge. No significant differences were observed between the 

two vaccinated groups for clinical signs and virus load parameters. 

Conclusion: Vaccination with N SRS partially protects calves against 

challenge with a wild bovine RSV strain. No differences were observed 

between the two vaccinated groups, suggesting that intranasal 

vaccination did not prime mucosal immunity in our experimental 

conditions. Studies are underway to improve the mucosal route of 

vaccination as well as immunogenicity of the candidate RSV vaccine. 

Key words: Respiratory Syncytial Virus, vaccination 

179 Efficacy of an Inactivated BTV-8 Vaccine against a Virulent 

BTV-8 Challange in Sheep 

C. Hamers 1 , M. Blanchet 1 , A. Chossat 1 , L. Besancon 2 , C. Cariou 2 , 

E. Hanotel 2 , S. Goutebroze 1 

1 MERIAL SAS, Clinical Development, Saint Vulbas, France 

1 MERIAL SAS, Bio-Analytical Development, Lyon, France 

Introduction: Bluetongue virus (BTV) causes an infectious, noncontagious, 

Office International Epizooties (OIE) and BTV cycle can 

be interrupted by the systematic vaccination of susceptible species, 

particularly sheep and cattle. Here, we demonstrated in a vaccination / 

challenge study that single administration of an inactivated vaccine 

containing purified BTV serotype 8 (BTV-8) was able to prevent 

clinical signs and viraemia in sheep. 

Material & Methods: Eleven weaned lambs were randomly allocated 

to one group of 5 vaccinates and one group of 6 control sheep. 

Vaccinates were subcutaneously treated once on day 0, with 1 mL of 


BTVPUR* AlSap8 (MERIAL). The other group was left unvaccinated 

and served as control. Thirty one days after vaccination, all sheep were 

challenged with a virulent BTV-8. All sheep were then monitored for 

rectal temperature and clinical signs from 5 to 14 days after challenge. 

Furthermore, viraemia of all sheep was monitored by quantitative RT- 

PCR on whole blood samples on days 5, 7, 9, 12 and 14 after challenge. 

Results: Hyperthermia : Vaccinates were significantly protected from 

hyperthermia. Average maximal hyperthermia was 41.2°C in the 

controls and 39.9°C in the vaccinates. Clinical scores: Vaccinates were 

significantly protected from clinical signs. Average global clinical 

score was 28.8 in the control group and 1.2 in the vaccinated group. 

Viraemia (qRT-PCR): All controls were found positive (moderate to 

high copy numbers) at all time points. None of the vaccinated animals 

was ever detected positive. 

Conclusion: In the present study, single vaccination with the product 

tested provided full and significant clinical and virological protections 

against a BTV-8 challenge. These results show that BTVPUR* AlSap 

8 may be used for BTV-8 disease prevention (clinical protection) and 

for epidemiological control of BTV-8 (virological protection). 

* BTVPUR AlSap is a registered trademark of the MERIAL group in 

the European Community. 

Key words: Bluetongue, BTV-8, inactivated vaccine, efficacy, sheep 

180 Efficacy of an Inactivated BTV-8 Vaccine Against a Virulent 

BTV-8 Challange in Cattle 

C. Hamers 1 , M. Blanchet 1 , F. Teppe 1 , L. Besancon 2 , A. Meyet 2 , 

S. Goutebroze 1 

1 MERIAL SAS, Clinical Development, Saint Vulbas, France 

2 MERIAL SAS, Bio-Analytical Development, Lyon, France 

Introduction: Bluetongue virus (BTV) causes an infectious, noncontagious, 

Office International Epizooties (OIE) and BTV cycle can 

be interrupted by the systematic vaccination of susceptible species, 

particularly sheep and cattle. Here, we demonstrated in a vaccination / 

challenge study that administration of an inactivated vaccine 

containing purified BTV-8 was able to prevent cattle from clinical 

signs and viraemia. 

Material & Methods: Ten cattle were allocated to 2 groups of 5 

animals. One group was subcutaneously vaccinated twice (day 0 and 

28) with 1 mL of BTVPUR* AlSap8 (MERIAL). The other group was 

left unvaccinated and served as control. On day 51, all animals were 

challenged with a virulent BTV-8. All cattle were then monitored for 

rectal temperature and clinical signs from 5 to 28 days after challenge. 

Furthermore, viraemia of all animals was monitored by quantitative 

RT-PCR on whole blood samples, three times a week during the 

monitoring period. 

Results: Hyperthermia: All controls presented an elevated rectal 

temperature, whereas rectal temperatures remained normal for all 

vaccinates. Clinical scores: Vaccinates were significantly protected 

from clinical signs. Average global clinical score was 59.0 in the 

control group and 10.8 in the vaccinated group.Viraemia (qRT-PCR): 

All controls were found positive (moderate to high copy numbers) at all 

time points but at day 56 none of the vaccinated animals was ever 

detected positive. 

Conclusions: In the present study, vaccination with BTVPUR* AlSap 

8 provided full and significant clinical and virological protections 

against a BTV-8 challenge. These results show that BTVPUR* AlSap 

8 may be used for BTV-8 disease prevention (clinical protection) and 

for epidemiological control of BTV-8 (virological protection). 

* BTVPUR AlSap is a registered trademark of the MERIAL group in 

the European Community 

Key words: Bluetongue, BTV-8, inactivated vaccine, efficacy, cattle 

181 Rapid Detection of Rift Valley Fever Virus Based on Viral 

Small (S) RNA Genome Segment 

I. Aradaib 1 , M. Al-dubaib 2 

1 

The National Ribat University, Medical Lab. Sciences, Khartoum, 

Saudi Arabia 

2 

Qassim University, Veterinary Medicine, Buridah, Saudi Arabia 

A reverse transcriptase Polymerase Chain Reaction (RT-PCR)-based 

assay was developed and evaluated for detection of Rift Valley Fever 

Virus (RVFV) ribonucleic acid (RNA). A pairs of oligoribonucleotide 

primers selected from the small (S) RNA genome segment of RVFV, 

were designed and used for PCR amplification. The primers resulted in

amplification of a primary 490 base pair PCR product. The PCR 

products were amplified from RNAs extracted from RVFV field 

isolates and vaccine strains, propagated in Vero cell cultures. 

Amplification products were not detected when the RT-PCR-based 

assay was applied on RNA from other related hemorrhagic fever 

viruses including Crimean Congo Hemorrhagic fever; dengue virus; 

epizootic hemorrhagic disease virus and total nucleic acid extracts from 

uninfected Vero cells. The described RT-PCR-based assay provides a 

rapid, sensitive and specific assay for detection of RVFV in cell culture 

and should be recommended for inclusion during outbreaks of the 

disease among humans and susceptible livestock. 

Key words: viral hemorrhagic fevers, RVFV, sRNA, RT-PCR 

182 Prevention of Coxiella burnetii Shedding in Intedted Dairy 

Herds Using a Monovalent Vaccine Containing Phase-1 

Coxiella burnetii 

R. Guatteo 1 , F. Beaudeau 1 , A. Joly 2 , D. Remmy 3 , H. Seegers 1 

1 

National Veterinary School of Nantes, Unit of Animal Health 

Management, Nantes, France 

2 

Union Bretonne des Groupements de Défense Sanitaire, Vannes, 

France 

3 

CEVA Santé Animale, Libourne, France 

Objective of study: The main aim of this study was to assess the 

efficacy of a monovalent vaccine containing phase I Coxiella burnetii 

to prevent Coxiella shedding in susceptible (ie considered as non 

infected) dairy cows within infected herds in comparison to a placebo. 

Materials and Methods: To reach this goal, 336 dairy cows and 

heifers, from 6 infected herds with repeated abortions due to Coxiella 

burnetii, were followed up over a one-year period. Before treatment (ie 

vaccination or placebo), the status of the cows towards Coxiella 

burnetii infection was determined on the basis on PCR results on milk, 

vaginal mucus and faeces and serological analysis performed 2 weeks 

apart. A cow was considered as susceptible when all PCR and 

serological results were negative, and was considered as infected 

otherwise. Then, the allocation of treatments was performed randomly 

within pregnant and non pregnant cows. After treatment (D0), the 

animals were subject to systematic samplings (milk, vaginal mucus and 

faeces) on D90, D180, D270 and D360 to detect a putative shedding 

using PCR. Moreover, under the assumption that calving period is at 

maximum risk of shedding, the same samples were taken within 15 

days after calving. An animal was considered as a shedder at a given 

time t, if at t, it was found positive-PCR tested at least once among the 

taken samples. The effect of treatment on the risk for an animal of 

becoming shedder was assessed using survival analysis techniques 

(Cox regression model). 

Results: Almost all heifers were detected as susceptible before 

treatment. When vaccinated while not pregnant, an animal had 0.2 

(P

635 Prevalence of the Pulmonary Lesions of the Bovines of the 

Area of Tiaret 

B. Mabrouk, T. Madjid, M. Toufik, M. Bakir 

Batna University, Department of Veterinary Sciences, Batna Algeria, 

Algeria 

The seizures partial or total of the carcasses or the bodies by the veterinary 

services constitute a true fear for the butchers. A study undertaken in the 

slaughter-houses of the wilaya of Tiaret (Western Algeria) during the last 

two years and a great incidence of respiratory lesions at the bovines was 

found. This study shows the prevalence and the frequency of a series of 

lesions raised on the lungs of 870 bovine autopsiés during medical 

inspections and 744 pathologies were diagnosed. The seasonal 

distribution of the lesions made it possible to put forward the effect of the 

season on the appearance of the lesions. The cyst hydatic constitutes 

pathology the most met (n=330 either 42.64%) followed 111 cases of 

emphysema (or 14.35%) and 61 cases of congestion (or 7.89%). The 

histopathologic study of the fragments of injured lungs taken at the 

slaughter-house highlighted tissue deteriorations. According to the 

localization of the lesions, one noted that the right lung is touched the most 

for 92.60% of the cases that the left lung as well as the craniaux lobes for 

87.05% comprativement with the caudal lobes of the lungs. 

Key words: pulmonary lesions, histopathology, bovine 

636 A Retrospective Study of Bovine Tuberculosis Eradication 

Project and Human Tuberculosis 

M. Sakha 1 , N. Ezatpanah 2 , A. Rohitoorbati 3 

1 Islamic Azad University, Science & Research Branch, Internal 

Medicine, Tehran, Iran 

2 National Research Center, Internal Medicine, Kerman, Iran 

3 Faculty of Veterinary, Internal Medicine, Kerman, Iran 

Introduction: Tuberculosis (TB) is a chronic and zoonotic disease, 

had known for many years. The cause of the disease is acid-fast bacillus 

of mycobacteriacea, mycobacterium tuberculosis. Weakness, 

emaciation and chronic respiratory symptoms are common signs of the 

disease. The main group of human TB is pulmonary TB and the other 

is extra pulmonary TB. It seems that many cases of extra pulmonary 

TB are related to animal tuberculosis. This is a retrospective study 

made during the last ten years (1991- 2000) on the incidence of human 

and bovine TB and evaluated the bovine TB control project to human 

TB incidence in city of Kerman, IRAN. Bovine TB control project is a 

national project that has performed for many years in IRAN. 

Materials and Methods: All data were taken from the veterinary 

office in Kerman for bovine TB control project and the only centre of 

human tuberculosis control for human cases. Data were analyzed and 

variables of age, gender, root of infection and social situation were 

considered in human section. The bovine positive cases have been 

omitted and sent to the slaughter house before. 

Results and Conclusion: This study showed that the most patients 

were over 60 years old and then 21 to 30 years old. Women showed 

more incidence of the disease. The prevalence of the extra pulmonary 

TB in the first year of the study was11.9 in 100,000 and two years later 

was relatively higher than. In remaining years the prevalence showed a 

decreasing slope and reached to half of early year. Among the 219375 

cows tested for TB in ten years,`65 cows were reactor. The most 

incidence was found in the first and second years, that is, 37 and 18 

cases, respectively. In later years, there was obviously a decreasing 

slope with only one or two reactors in 1993 to 1998 and no cases in 

1999 and 2000.There is positive relation between decrease in human 

TB (extra pulmonary TB ) and the control of bovine TB in Kerman, so 

that with the obvious decreasing of bovine reactors in last year of this 

study, human cases also had a decreasing slope, although the human 

extra pulmonary TB is nearly one third of total human cases and it 

needs supplemental projects to control of human tuberculosis. 

Key words: tuberculosis, bovine, human, retrospective 

637 Bovine Theileriosis: Evaluation of 372 Cases during Spring 

and Summer in South of Iran 

M. Sakha 1 , M. Radfar 2 , M. Janbaz 3 

1 Islamic Azad University, Science & Research Branch, Internal 

Medicine, Tehran, Iran 

2 Faculty of Veterinary Medicine, Parasitology, Kerman, Iran 

3 Faculty of Veterinary Medicine, Internal Medicine, Kerman, Iran 


Bovine theileriosis caused by Theileria annulata is a dangerous and 

important disease in tropical and subtropical areas. There are large 

economical losses in these areas every year including Iran. The vector 

of the disease in Iran, the tick of Hyalomma is found in the most parts 

of the country. The disease is more seen in spring and summer. In this 

study, the prevalence of the disease in terms of month of the year, age, 

sex and animal breed were studied. Results showed that the maximum 

and minimum prevalence of the disease was in May and June, 

respectively, during the studied months. In relation to age, 76.2 percent 

of the infected cattle were under a month old. In relation to the breed, 

infection of different breeds of Holstein, crossbred and native cattle 

were 1.6, 94.4 and 4.0 percent, respectively. In relation to sex, 55.4% 

of males and 44.6% of females were affected. From the aspect of 

clinical signs, nearly all cattle showed anorexia and enlargement of 

lymph nods. 58% of blood smears showed less than 10% 

intraerythrocytic form of theileria in microscopic high power field, 

38% showed more than 10 percent and the rest was uncertain to 

diagnose. It is concluded that in spite of all the means of prevention and 

vaccination, the disease has been known as the most important disease 

in the region especially in the warm months of the year parallel to the 

activity of hyalomma. The dominant affected breed was crossbred, 

because the most of the referred animals was crossbreds. Calves were 

the most affected in this study because most of our cases were calf and 

cattle were more treated at the farms. The infected RBC were not 

correlated to the severity of the disease. 

638 New Approaches in the Prophylaxis of Prion Infections 

V. Vlizlo, P. Verbitskii, C. Mayor, V. Stadnyk 

Institute of Animal Biology UAAS, Scientific Center for Prion 


Objectives of study: In nowadays very much the attention is spared to 

the problem of the treatment of prion infections. Most efforts are 

concentrated on the search of preparations which would block 

occurance or reduce the level of already present PrP SC . At the same 

time, very little attention is spared to the problem of prophylaxis of the 

prion infections. Although work in this direction is rather active, taking 

into considiration a creation of transgenic cattle in which the gene of 

Prnp is knocked-out. But still it is unknown, how exactly will react an 

organism on absence of such important protein as PrP C . That is why we 

consider, that more perspective direction is a search of preparations 

which would reduce the level of PrP C , in the same time depriving PrP SC 

of substrate for replication. These preparations can be applied in risky 

herds, with suspicious or sick animals are founded, for the reduction of 

risk of prion infections among the healthy individuals. That is why the 

purpose of our work was a search of preparations which would reduce 

the level of PrP C in the organs of the prion-replicating system of 

laboratory animals. 

Materials and Methods: Work was done on laboratory rats treated 

with heparine and Petosan Polisulfate. After decapitation in the proper 

organs were probed: general maintenance of PrP C , type of expression 

of its izoforms, maintenance of zinc and copper, activity of 

superoxidedismutase, evaluation of DNA damage, ELISA test of PrP C 

interaction with pentosan polisulfate (PPS), determination of UVspectrums 

of PrP C at interaction with PPS. 

Results: It is set that heparine and PPS is able to reduce the level of 

PrP C in the organs of the prion-replicating system; unlike heparine, it is 

shown that PPS operates in far less concentrations and does not 

influence on the state of the antioxidant system. It is also shown that 

action of PPS on PrP C proportionally depends on the concentration of 

preparation. In the case of heparine such dependence was not 

discovered. It is set that both preparations do not influence on genomic 

DNA, which is proved by the absence of doblestrain DNA breaks. 

Conclusions: It is set an inhibiting action of PPS and heparine on PrP C 

in vivo. 

Key words: prion infections, cellular prion, pentosan polysulphate 

639 Brucellosis Surveillance in Jordan, 2001-2006 

B. Barqawi 1 , Z. Mashhadani 1 , B. Hijjawi 2 

1 Central of Disease Control & Prevention-CDC, Field Epidemiology 

Training Program-FETP, Amman - Jordan, Jordan 

2 MOH, DCD, Amman - Jordan, Jordan 

Background: Brucellosis is a zoonotic bacterial disease that often 

infects livestock animals (sheep, goats, and cattle). Animals often shed

organisms during birth. Humans become infected by coming into 

contact with animals or animal products contaminated with the 

bacteria. The objective of this study was to examine brucellosis 

surveillance and livestock immunization data in order to make 

recommendations that can improve brucellosis prevention and control 

programs in Jordan. 

Methods: All health directorates in Jordan electronically report the 

number of cases of confirmed and probable human brucellosis to the 

Ministry of Health weekly. Confirmed human cases of brucellosis are 

diagnosed by standard Coombs tube agglutination test. Clinically 

compatible cases with supportive serology (Rose Bengal test) and 

epidemiologically linked to confirmed cases are defined as probable 

cases. The Veterinary Directorate, Ministry of Agriculture collects data 

on livestock and immunization. The study period was 2001 to 2006. 

Results: 1,137 cases of human brucellosis were reported in Jordan 

during 2001 to 2006. The mean yearly number was 190. Mean yearly 

human brucellosis incidence was highest in Mafraq (30/100,000 

population) and Karak (12) Governorates. Mafraq (4:1) and Karak 

(3:1) have the highest mean animal-to-human density of sheep and 

goats in Jordan. Human cases vary by month of occurrence; most cases 

(651; 57%) were from May to August and few cases (214; 19%) were 

from October to February. Immunization of sheep and goats in Jordan 

increased from 15% in 2002 to 46% in 2006. 

Conclusions: Human cases of brucellosis cluster geographically in areas 

with high density of sheep and goats. Occurrence of human cases parallels 

the spring-summer birthing pattern in animals. Human brucellosis is 

highly underreported. WHO burden of disease studies estimate there may 

be 3,000 human cases a year in Jordan. There does not appear to be any 

recent change in the incidence of human brucellosis or animal disease. 

New surveillance methods to identify more cases will be explored, such as 

following up recipients of antibiotic prescriptions typically used in 

brucellosis treatment. The Ministry of Agriculture and Ministry of Health 

increased immunizations to almost 2 million animals in 2006, but 

effective prevention will require sustained efforts to maintain high 

immunization levels, particularly in Mafraq and Karak. 

640 Comparison of Biochemical Profiles of Hydatid Cyst Fluids of 

Echinococcus granulosus from Different Hosts in Iran 

A. Eslami 1 , P. Khazraii 2 , M. Heidarpour 2 , Hr. Mohamadi 2 


of Parasitology, Tehran, Iran 



Hydatidosis (echinococcosis) is caused by the cystic larval. Stage of 

Echinococcus granulosus, is recognized as being one of the world’s 

major zoonoses. Hydatidosis is endemic in Iran where many domestic 

animals including sheep, goat, cattle and camel act as intermediate 

hosts, while dog coyote and wolf act as final host of E. granulosus. 

There are nine host adapted strains of E. granulosus. The strain 

characterization is particularly important in regions where more than 

one species of live stock intermediate host exist and where there is the 

possibility of different cycles of transmission and source of infection 

for humans. Biochemical studies are useful in differentiating strain 

variations of E. granulosus in different countries. 15 sample of hydatid 

fluids were collected from the lung and liver of each host (i.e. sheep, 

cattle and buffalo).The concentrations of triglycerids, cholesterol, 

glucose, urea, creatinine, uric acid, total protein, magnesium, calcium, 

phosphorous, LDH, ALP, AST, CPK, GGT, Na and K + were 

determined. The biochemical profiles of lung and liver in these hosts 

were compared with each other. Hydatid fluids from the lung cysts of 

sheep contained significantly more glucose, uric acid, magnesium and 

LDH than those of cattle; but there were no significant differences on 

the concentration of other factors in hydatid fluids from lung cysts of 

these two species. There were other similarities and significant 

differences in the biochemical composition in hydatid cyst fluids of 

different hosts which can assist in the characterization of the host origin 

of infectivity in human being. 

641 Vaccinia Virus in Humans and Cattle in Southwest Region of 

Sao Paulo State - Brazil 

J. Megid 1 , C. Appolinario 1 , R. Carreira 1 , H. Langoni 1 , M. Pituco 2 , 

L. Okuda 2 

1 UNESP- School of Veterinary Medicine and Animal Sciences, 

Veterinary Hygiene and Public Health, Botucatu, Brazil 

2 

Instituto Biológico, Laboratorio de Viroses de Bovideos, Sao Paulo, 

Brazil 

Although human vaccine was erradicated in the world and the 

vaccination programs stopped, outbreaks of Vaccinia Virus has been 

reported in several states and regions of Brazil from which different 

types of Vaccinia Virus (VV) were isolated and characterized. 

Recently, Trindade et al (2007) studied the origin and diversitiy of 

Brazilian Vaccina Virus isolated evaluating the possibility that those 

could represent an escaped vaccinal strain. The authors reinforce the 

persistence of vaccinia virus in Brazil and other parts of the world 

despite the smallpox eradication campaign and the concept that the 

vaccinal strain could no more exist in nature. 

Objective: In this paper we report another cases of Vaccinia Virus 

infection in cattle and humans in the southwest region of Sao Paulo state 

in Brazil. The disease was observed in several small dairy farms with 

manual milking. Lesions varying from vesicules to ulcers were observed 

in the teats of the cattle and mouth of one calf. Milkers presented lesions 

in the hands and one child, 11 years old, presented lesions in mouth and 

nose. The age of the milkers ranged from 47-63 years old. 

Material and Methods: Crusts and swabs were collected from the 

animals that presented lesions in initial and healing stages.Crusts and 

swabs were maintained in Eagle Minimal Essential Medium (MEM) 

with penicilyn (500U/mL) and mycostatin (10 µg/mL) and maintained 

in refrigeration and sera were freezed. The material was processed for 

virus isolation and PCR assays. 

Results: A Vaccinia Virus was isolated in cell culture from all crusts 

and confirmed by PCR assay. 

Conclusion: Outbreaks of Vaccinia Virus in cattle in Brazil has been 

frequently reported. Lesions are observed in animals as well as in 

humans older than 40 years previously vaccinated against variola. This 

paper reports the characterization of new focus of Vaccinia Virus in 

Brazil and reinforce the susceptibility of humans previously vaccinated 

against variola to the circulating strain of vaccinia virus. 

Key words: Vaccinia Virus, cattle, human, outbreaks, public health 

642 Mycoplasma bovis Sanitation Program in a Dairy Herd using 

Baytril 100/Max and Biosecurity Measures 

D. Gevaert 1 , P. Passchyn 2 

1 Bayer SA-NV, Animal Health Division, Brussels, Belgium 

2 DAP De Toren, Veterinary Medicine, Torhout, Belgium 

Objectives of the study: A sanitation program in a dairy herd with 

bovine respiratory disease (BRD) initiated by Mycoplasma bovis was 

tested. 

Materials and Methods: A dairy herd of 117 Holstein Friesian cattle 

suffered since 2001 from BRD: relapses and growth retardation 

(calves) + chronic coughing (adults). Tilmicosin and tulathromycin 

treated calves relapsed after 14 days. Calves are housed individually up 

to 2 weeks, followed by group housing in 3 age groups (1, 2, 3 months) 

in the same stable. The calves are vaccinated against IBR, Pasteurella 

haemolytica, Parainfluenza and BRSV. Sanitation program: Step 1: 

herd visit + analysis of the spread of M. bovis using ELISA serology. 

Step 2: enrofloxacin (Baytril 100/Max) treatment + biosecurity 

measures. Step 3: follow-up visits. 

Results: Step 1: the M. bovis antibody prevalence increased with age: 

25% in calves

Conclusions The sanitation program reduced the antibiotic cost by 

50%, gave less BRD signs and 29% less seropositive animals. The 

program must be continued until the M. bovis herd prevalence has 

dropped to an acceptable level. 

Key words: Mycoplasma bovis, enrofloxacin, biosecurity 

643 Analysis of Pathways for Release of Rift Valley Fever Virus 

into the Continental United States 

R. Kasari 1 , A. Carr 2 , V. Lynn 3 , T. Weaver 3 

1 

USDA-APHIS-VS-CEAH, National Surveillance Unit, Fort Collins, 

United States 

2 

Uniformed Services University of the Health Sciences, Department 

of Preventive Medicine and Biometrics, Bethesda, United States 

3 

USDA-APHIS-VS-CEAH, Center for Emerging Issues, Fort Collins, 

United States 

Objectives of Study: The primary objectives of this study were to evaluate 

the feasibility of RVFV to enter the continental United States by various 

routes as well as identify the states whose domestic and wild ruminant and 

human populations are most vulnerable for exposure to this virus. 

Materials and Methods: Pathways analysis methods were used to 

identify the pathways for release of RVFV into the continental United 

States. Initially, the agent, host, and environmental factors important in 

the epidemiology of RVF were used to develop a list of potential 

pathways for release of RVFV into the continental United States. Five 

pathways were identified: (1) importation of RVFV-infected animals, 

(2) entry of RVFV-infected people, (3) mechanical transport of RVFVinfected 

insect vectors, (4) intercontinental wind-borne transport of 

RVFV or RVFV-infected insect vectors, and (5) smuggling of live 

virus. Next, the feasibility of each pathway was evaluated using data 

from governmental and public domain sources to evaluate animals, 

commodities, and people that moved from RVF-endemic countries to 

the continental United States between calendar years 2000 and 2005. 

Finally, entry points of each feasible pathway into the continental 

United States were used to identify the domestic and wild ruminant and 

human populations at risk for exposure to RVFV. 

Results: Feasible pathways for entry of RVFV into the continental 

United States are (1) importation of RVFV-infected animals, (2) entry 

of RVFV-infected people, (3) mechanical transport of RVFV-infected 

insect vectors, and (5) smuggling of live virus. 

Conclusions: A pathways analysis provides the requisite information 

needed to construct an effective targeted surveillance plan for this virus 

in order to enable rapid detection and response by animal health and 

public health officials. Domestic ruminant livestock, ruminant wildlife, 

and people in 14 states (Alabama, California, Florida, Georgia, Maine, 

Maryland, Massachusetts, Minnesota, New Jersey, New York, 

Pennsylvania, South Carolina, Texas, and Virginia) appear to be most 

vulnerable to exposure from RVFV. It should be noted however, that 

this group of “at-risk” states could be altered entirely, if this viral agent 

is released illicitly as part of a terrorist act. 

Key words: Rift Valley Fever, zoonotic disease, pathways analysis 

644 Analysis of Pathways for Release of Ehrlichia ruminantium 

(Heartwater) into the Continental United States 

R. Kasari 1 , E. Lawrence 2 , H. Howe 2 , L. Farnsworth 2 , E. Freier 2 , 

M. James 2 , S. Miller 2 

1 

USDA-APHIS-VS-CEAH, National Surveillance Unit, Fort Collins, 

United States 

2 

USDA-APHIS-VS-CEAH, Center for Emerging Issues, Fort Collins, 

United States 

Objectives of Study: The objective of this study was to evaluate the 

feasibility of Ehrlichia Ruminantium by itself or via E. Ruminantiuminfected 

Amblyomma spp. vector ticks to enter the continental United 

States by various routes as well as identify the States whose domestic and 

wild ruminant populations are most vulnerable for exposure to this 

rickettsial organism. 

Materials and Methods: Pathways analysis methods were used to 

identify the pathways for release of E. ruminantium into the continental 

United States. Initially, the agent, host, and environmental factors 

important in the epidemiology of E. ruminantium infections in susceptible 

animal species were used to develop a list of potential pathways for 

release of this rickettsial agent into the continental United States. Five 

pathways were identified: (1) importation of E. ruminantium-infected 

animal species, (2) migrating cattle egrets (Bubulcus ibis) serving as a 


transport host for E. ruminantium-infected Amblyomma spp. ticks, (3) 

mechanical transport of E. ruminantium-infected Amblyomma spp. ticks 

by humans and imported non-ruminant animals, reptiles, and other birds, 

(4) mechanical transport of E. ruminantium-infected Amblyomma spp. 

tick by fomites, and (5) smuggling of live E. ruminantium. Next, the 

feasibility of each pathway was evaluated using data from governmental 

and public domain sources to evaluate animals, commodities, and people 

that moved from heartwater-endemic African and Caribbean island 

countries to the continental United States between calendar years 2000 

and 2006. Finally, entry points of each feasible pathway were used to 

identify the States whose domestic and wild ruminant populations were at 

greatest risk for exposure to E. ruminantium. 

Results: All five pathways were feasible ones for entry of E. 

ruminantium or E. Ruminantium-infected Amblyomma spp. vector ticks 

into the continental United States. 

Conclusions: Compared to other states, ruminant livestock and 

ruminant wildlife species in 17 states (Alabama, California, Florida, 

Georgia, Illinois, Louisiana, Massachusetts, Maryland, Minnesota, 

Mississippi, North Carolina, New Jersey, New York, Pennsylvania, 

South Carolina, Texas, Virginia) should be most vulnerable to exposure 

to Amblyomma spp. ticks, some of which could be infected with E. 

ruminantium. This group of “at-risk” states could be altered entirely, if 

the rickettsial agent is released illicitly as part of a terrorist act. 

Key words: heartwater, Ehrlichia ruminantium, pathways analysis 

645 Epidemic of Bovine Viral Diarrhea Virus (BVDV) Infection 

Originated from 3 Persistently Infected Cattle on a Farm 

during a 5-year Period 

M. Tajima 1 , K. Sano 2 

1 Hokkaido University, Veterinary Teaching Hospital, Sapporo, Japan 

2 NOSAI Iburi, West branch, Toya, Japan 

Bovine viral diarrhea virus (BVDV) is distributed all over the world. 

Persistently infected cattle with BVDV (PI) are a latent source of infection 

of BVDV in the herd, and can cause serious economic damage. Testing 

and culling of PI are important procedures for dairy production. We 

encountered 3 outbreaks of BVDV infection in a farm during a 5-year 

period, and these were suspected to have caused by 3 individual PI. First 

epidemic: In May 2002, a 6-month-old Holstein calf was diagnosed as PI 

because of mucosal disease. A sister cow (primipara) of the PI calf and 

their dam (PI-1) were diagnosed as PI, and the bulk tank milk (BTM) test 

was positive for BVDV. After exclusion of all PIs, BTM test became 

negative for BVDV. The negative results were obtained twice at an 

interval of 6 months, therefore, the test of BVDV was discontinued. 

Second epidemic: In January 2005, 2 calves (pneumonia and poorgrowth) 

were diagnosed as PI. Their dams were BVDV negative and 

BTM test was positive again for BVDV. All cattle on the farm were 

examined for BVDV. Six PIs were detected, a milking cow imported from 

the cattle market in May 2004 (PI-2), 4 Holstein calves and a beef calf. By 

examination of all neonatal calves thereafter, 14 PIs were identified during 

a 6-month period. During this period, 50 milking cows were imported into 

the farm. All of these animals were BVDV negative, but 5 calves 

produced from these cows were diagnosed as PI. Except for these 5 

calves, 9 calves and PI-2 were infected with the same BVDV based on the 

analysis of E2 diversity of the isolated BVDV, however, the BVDV 

differed from that involved in the first epidemic on the farm. After 

exclusion of all PIs, there were no PIs for a 3-month period. Third 

epidemic: In October 2005, an imported dam (PI-3) and neonatal calf 

were diagnosed as PI. At this time, the importation of cattle was 

discontinued, and the examination of newborn calves for BVDV was 

continued. After 6 months, 2 PIs were detected. All 4 PIs were infected 

with the same BVDV, and BVDV from PI-3 differed from those of PI-1 

and -2. After the final detection of PI, approximately 250 newborn calves 

were examined for PI during a one-year period (till May 2007). There 

were no PIs and BTM test was negative every 6 months; therefore, the 

farm was recognized as BVDV free. Continuous BTM test and 

examination of newborn calves are the most important and effective 

procedures for BVDV eradication. 

Key words: BVDV, eradication, test and cull 

646 Paraclinical Findings of Chronic Extensive Peritonitis in a 

Diarrheic Young Calf 

N. Atyabi, N. Alidadi, M. Hashemian 

Faculty of Veterinary Medicine, University of Tehran, Animal 

Science, Tehran, Iran

A 5-month-old calf with a chronic diarrhea and bloat and obvious 

abdominal pain was referred to the large Animal Teaching and 

Research Hospital. On physical examination the calf was lethargic and 

had a rectal temperature of 39.8 ˚C, respiratory rate 44 breaths per 

minutes, heart rate 100 beats per minutes and mild tachycardia. The 

calf had pale mucosal membranes and clinical dehydration. The 

peritoneal fluid examination showed an exudate with high protein level 

and degenerated neutrophil which was positive for Salmonella dublin. 

The fecal samples were negative for parasites and Mycobacterium 

paratuberculosis, but positive for Salmonella dublin. The leukogram 

showed significant leukocytosis, lymphocytosis and monocytosis 

accompanying by neutropenia and a shift to left. A degenerative 

anemia was diagnosed in the calf which could be a ambiguous 

prognosis. The animal died despite of relevant fluid and antibiotic 

therapy, and a chronic extensive peritonitis was found in the 

postmortem examination 

Key words: calf, chronic extensive peritonitis, diarrhea, leukogram 

647 Molecular Survey on Occurrence of Mycoplasma Mycoides 

Subspecies Mycoides L.C and S.C in Pulmonary Infection of 

Iranian Cattle, Goats and Sheep 

N. Atyabi, Z. Nikusefat, SA. Pourbakhsh 



This survey described microbiological and molecular study for detection 

Mycoplasma mycoides subspecies mycoides (L.C and S.C) in pneumonic 

lungs of Iranian sheep, goats and cattle. Suspected samples were 

investigated based on color, consistency and appearance of the cut 

surface. Total 180 lung samples were collected from slaughter house 

located in West of Iran, Kermanshah during 2006-2008 and transferred 

near ice packs to Razi Research Institute. Gross lesions showed 

hepatization with grey and white lesions (consolidation) and motley 

appearance with or without fibrin. Minced lung tissues were inoculated to 

PPLO broth agar (Diffco) with application acetatecellulose filter (0.45-2.5 

micrometer). Samples were in 6.8% carried to Co2 incubator. After 10-14 

days the yellow tubes were subcultured to PPLO plate. After while, 

characteristic colony was observed. DNA extractions were based on 

phenol method and DNA extracted isolates were preserved in 50% 

glycerol at -20˚C.DNA extract of all samples were subjected to generic 

and species specific PCR based on 16 S rRNA with different set of 

primers and cycles. The visualized amplicon consisted of 573 bp.40 

samples from total 180 lung extract were positive for genus mycoplasma 

(22.2%),while only 30 samples were positive in culture(16.66%) .There 

were no significance difference in sex and age between affected 

animals.(P>0.05) The highest percentage of infection was observed 

during December (32%) and the lowest was in June (2.5%).There were no 

evidence of Mycoplasma mycoides mycoides (S.C ) and (L.C) by species 

specific primer based on CAP-21 PCR in infected ruminants. However, 

the present work was carried out to study the incidence of two strains of 

mycoid cluster in Iranian ruminants. Further investigation should be 

conducted to profile all mycoplasma spp in pulmonary infections. 

Key words: Mycoplasma mycoides, sub. mycoides (L.C and S.C), 

ruminant, Iran 

648 Application of PCR and Hematological Findings in 

Replacement Dairy Heifers with ELISA Seropositive to 

Bovine Leukosis Virus Infection 

T. Rukkwamsuk, S. Panneum 

Faculty of Veterinary Medicine, Kasetsart University, Large Animal 

and Wildlife Clinical Science, Nakhon-Pathom, Thailand 

Bovine leukosis virus infection was studied in 171 replacement dairy 

heifers. Blood samples were collected for determination of 

hematological parameters, detection of bovine leukosis virus using 

PCR technique, and determination of antibody against bovine leukosis 

virus infection using indirect ELISA (IDEXX HerdCheck Anti-BLV). 

Blood samples from seropositive and seronegative heifers were 

randomly selected to detect bovine leukosis virus using PCR technique. 

Results revealed that seroprevalence of bovine leukosis virus infection 

in this group of heifers was 19.3% (33 heifers were seropositive and 

138 heifers were seronegative). Most hematological parameters did not 

differ between seropositive and seronegative heifers. Average total 

white cell counts were higher for seropositive heifers than for 

seronegative heifers (P = 0.08). Regarding PCR results, bovine 

leukosis virus was not detected from some seropositive heifers but 

could be detected from some seronegative heifers. However, results 

obtained from indirect ELISA and from PCR technique had a moderate 

agreement (Kappa value = 0.60). In conclusion, seroprevalence of 

bovine leukosis virus infection in replacement dairy heifers was 

relatively high. Increased total white cell count seemed to be related to 

bovine leukosis virus infection. Although agreement of serological test 

using ELISA and virus detection using PCR was moderate, results of 

these tests were still contradicted in some heifers. Therefore, further 

study on increasing sensitivity and specificity of diagnostic tests is 

required. 

Key words: Bovine Leukosis Virus, hematology, replacement heifer 

649 Bovine Leptospirosis in Sardinia: Isolation and Evaluation of 

Molecular Methods 

M. Ponti, G. Sanna, G. Carboni, G. Canu, M. Manca, M. Noworol, B. 

Palmas, E. Marongiu, C. Patta 

Istituto Zooprofilattico Sperimentale Della Sardegna, Sassari, Italy 

Previous studies carried out in Sardinia have shown that Leptospirosis 

affects both humans and farm animals and that Leptospira serovar 

pomona is responsible for the icterus-haemorrhagic syndrome in 

calves. In this study, the microscopic agglutination test (MAT) was 

used to determine the serological titers of the serum samples collected 

from cattle. Even though definitive diagnosis is provided by culture of 

pathogenic Leptospira, several molecular approaches were also 

evaluated, aiming at optimizing the detection of bacteria in bovine 

samples of urine and blood. Specimens were collected from two dairy 

cattle herds in the north of Sardinia. Farm A consisted of 580 subjects, 

out of which 260 were lactating cows, 280 were freerange heifers and 

40 were calves. Farm B included 120 animals, out of which 45 were 

lactating cows and the remaining ones were heifers and calves. Sera 

were examined by MAT. Urine, blood and tissue suspension were 

inoculated in semi-solid medium for cultivation and tested by PCR. 

The Leptospires strains isolated were identified by using monoclonal 

antibodies. Several primer pairs for detection of leptospiral DNA were 

tested: one set amplified a fragment of the 16S rRNA, another set was 

complementary to a portion of Lig protein gene (Lig 1/Lig 2) and a 

third one used G1/G2 and B64I/B64II. Then a protocol of real-time 

PCR using a Taqman Probe was standardized and compared with the 

three protocols previously described. In farm A the seroprevalence was 

of 60%, while in farm B it was about 40%. Leptospires were isolated 

from 8 and 3 urine samples in farms A and B, respectively. All strains 

were identified as Leptospira serovar pomona. Leptospires were not 

isolated from blood and fetuses samples. All the molecular methods 

used amplified leptospiral DNA from all 15 serovars of our Leptospires 

panel, and also from blood, urine and fetuses tissues. Primers derived 

from rRNA gene sequence were the least specific, and none of the 

molecular methods tested was 100% sensitive. Furthermore, the main 

limitation of these PCR-based assays was their inability to identify the 

infecting serovar. Real-time PCR assay, which successfully detected 

leptospiral DNA, is rapid and specific but more expensive than 

conventional PCR. A combination of two detection methods (PCR and 

culture) is the most sensitive approach for early diagnosis of 

Leptospirosis. 

Key words: leptospirosis, bovine, PCR 

650 Serological Surveillance of Contagious Bovine 

Pleuoropneumonia between 2005 and 2007 Years in Romania 

R. Radulescu, G. Petriceanu, A. Ragalie, E. Gutu 

Institute for Diagnosis and Animal Health, Immunology, Bucharest, 

Romania 

The reference method recommended by OIE for serological 

surveillance of Contagious Bovine Pleuropneumonia (CBPP) is the 

Complement Fixation Test (CFT). This method had been used in the 

past for eradication of CBPP in many countries. However, CFT have 

some disadvantages, mainly the production and standardization of 

specific antigen, providing some false positive reaction due to the 

mycoplasmas cross-reactivity from M. mycoides cluster. In Romania 

between 2005 and 2007, years have been examined for CBPP by CFT 

5306 samples taken from cattle, deer, wild sheep, reindeer and buffalo. 

The samples were analyzed with two CFT antigens Mycoplasma 

mycoides subsp. mycoides provided by CBPP OIE Reference 

Laboratories from LINV Lisbon-Portugal and ISZ G.Caporale Teramo- 


Italy. After the sample examinations, 90 (1.69%) were positive, 146 

(2.75%), doubtful, 77 (1.45%) anti-complementary and 4993 (94.13%) 

were negative. At the reanalyzing of those positive, doubtful and anticoplementary 

samples after a short period of time all of them were 

negative. The specificity of CFT had been calculated using both 

antigens and the following results were obtained: 95-99% with 

Portuguese antigen and 82-100% with Italian antigen. From the 649 

CFT examined samples using the Italian antigen which revealed a 

specificity of 82%, we examined by competitive ELISA (cELISA) all 

the positive (20), doubtful (85), anticomplementary (6) and 59 negative 

samples. From 170 examined samples only 12 presented doubtful 

results and the others were negative. For the introducing in the routine 

work of cELISA we also evaluated the precision parameter of the 

method by calculation of the repeatability and reproducibility of the 

obtained results. The variation coefficient (CV) of the reproducibility 

of control sera obtained results had been between 6.3% and 6.8% in 

comparison with 8% CV reported by the producer. Taken into 

consideration that cELISA had revealed a very good specificity of 92% 

and a correlation index of 41.76% against CFT results it is 

recommended that any anticomplementary, doubtful or positive results 

in CFT to be also analyzed by cELISA. 

Key words: M. mycoides, CFT, ELISA, CPBB, antigen 

651 Compatibility of Inactivated Bovine Viral Diarrhoea (BVD) 

and Leptospira Interrogans Serovar Hardjo Vaccines 

I. Mawhinney 1 , B. Makoschey 2 

1 ICM Broadlands, Bury St. Edmonds, United Kingdom 

2 Intervet-Schering Plough, Int. Marketing, Boxmeer, The Netherlands 

Objectives: The purpose of this study was to examine whether concurrent 

administration of a Leptospira vaccine affects the immunogenicity of an 

inactivated bovine viral diarrhoea (BVD) vaccine. 

Materials and Methods: Twenty beef breeding animals were included 

in the trial. At the beginning of the study, one animal had very low 

antibody titers against BVDV the remaining animals were 

seronegative. All animals were seronegative for Leptospira prior to the 

vaccinations. All animals were vaccinated intramuscularly with a 

commercially available BVD vaccine (Bovilis ® BVD, Intervet). Ten of 

them (group A) were vaccinated subcutaneously at the same time but 

on the opposite site of the neck with an inactivated Leptospira 

interrogans serovar hardjo vaccine (Leptavoid ® H, Schering Plough). 

The remaining 10 animals (Group B) were vaccinated two weeks later 

with the Leptospira vaccine following the manufacturer 

recommendation. The BVD vaccination was repeated after four weeks 

in both groups. The animals of group A received a second dose of the 

Leptospira vaccine at the same time. The animals of group B were revaccinated 

with the Leptospira vaccine 2 weeks after the BVD booster. 

Blood samples were taken at the first and second BVD vaccination 

(days 1 and 28) and two and four weeks after the second vaccination 

(days 42 for group A and 42 and 56 for group B). The samples were 

processed into serum for antibody testing. Antibodies specific for BVD 

virus were measured by serum neutralisation test. Leptospira serology 

was carried out against each of the two hardjo serovars, prajitno and 

bovis using a standard MAT test (VLA, Weybridge). 

Results: There was no statistically significant difference when the 

serological response to the BVD vaccine was compared between the 

two groups. Likewise, there was no statistically significant difference 

in the Leptospira serology, neither for prajitno, nor for bovis when the 

antibody responses two weeks after the second dose of the Leptospira 

vaccine was compared between the two groups. 

Conclusions: From these data it was concluded that there was no 

negative effect on the neutralising antibody response when the two 

vaccines were applied concurrently (at the same time but on opposite 

sides of the animal). 

652 Simultaneous Infection of Calves with Maternal Antibodies 

with Bovine Rotavirus and Coronaviruses under 

Experimental Conditions 

U. Schmidt 1 , M. Timmermans 1 , P. Tonen 1 , B. Makoschey 2 

1 

Intervet-Schering Plough, R&D, Boxmeer, The Netherlands 

1 

Intervet-Schering-Plough, Int. Marketing, Boxmeer, The 


Objectives: Newborn calf diarrhoea is generally recognized as 

multifactorial disease. The list of pathogens that can be involved is 


quite extensive, comprising bovine rotaviruses, bovine coronaviruses, 

E. coli and Cryptosporidium parvum as the most important ones. In the 

present study, clinical signs after simultaneous infection of newborn 

calves with two rotaviruses and a coronavirus under laboratory 

conditions should be compared to the infection with either of the three 

pathogens alone. Moreover, it should be investigated, whether all three 

viruses replicate after co-infection. 

Materials and Methods: The experiment was divided in four separate 

studies. On days 5, 6 and 7 after birth, the calves were infected orally 

with bovine rotavirus P5G6 (study 1), bovine rotavirus P11G10 (study 

2) and bovine coronavirus DB-2 (study 3) or all three viruses together 

(study 4). After the infection, the calves were observed twice daily for 

clinical signs of diarrhoea. From the day of infection onwards, fecal 

samples were taken twice daily. Virus shedding was quantified by 

ELISA technique. 

Results: All 12 animals infected with a single virus survived until the 

end of the observation period and some animals developed only very 

mild or no diarrhoea. By contrast, all four animals infected 

simultaneously with all three viruses had liquid faeces during at least 

one observation moment and two of the four animals had to be killed on 

humane reasons after episodes of severe diarrhoea. Both, rotavirus and 

coronavirus was detected in the faeces of all four animals infected 

simultaneously with the three viruses during several days. Moreover, 

both rotavirus G-types were present in a faecal sample from a coinfected 

calf. 

Conclusions: From these data, it can be concluded, that the coinfection 

of calves with rota- and coronaviruses under laboratory 

conditions reflects the field situation, where co-infection of 

pathogens causes more severe disease than infection with single 

pathogens. Moreover, the results reported here confirmed, that 

colostrum from unvaccinated dams did only partly protect against 

diarrhoea. Experimental data and field experiences have 

demonstrated the benefit of cow vaccination against enteric 

pathogens. 

653 Serum-free Produced Live Virus Vaccines are Efficacious and 

Safe 

B. Makoschey 1 , P. Gelder 2 


2 Intervet-Schering Plough, R&D, Boxmeer, The Netherlands 

Objectives: Most cell culture based manufacturing processes for 

animal and human vaccines use bovine fetal serum (FCS). In potential, 

FCS can be contaminated with a number of viruses that can infect the 

bovine foetus transplacentally, especially with the bovine viral 

diarrhoea virus (BVDV). Therefore, we have developed a novel 

production procedure for bovine herpesvirus type 1 (BoHV-1) live 

virus vaccines that does not make use of serum or fractions thereof 

during any cell or virus passage, thus completely serum-free. It should 

be determined whether serum-free produced BoHV-1 viruses are 

suitable as live vaccines for use in cattle. 

Materials and Methods: The safety of serum-free produced BoHV- 

1 virus was established in two separate studies. In the first study, six 

colostrum deprived calves each were vaccinated intranasally (IN) or 

intramuscularly (IM) with an overdose and two weeks later again 

with a single dose. In a second study, ten seronegative calves at the 

age of five to six months were vaccinated four times at fortnightly 

interval with an overdose, both IN and IM. The efficacy was tested 

in a vaccination-challenge study in seronegative calves as laid down 

in the European Phamacopeia. For comparison, a vaccine batch with 

BoHV-1 grown in serum-containing cell culture medium was 

included in the study. The calves were monitored for clinical signs 

after challenge infection with BoHV-1 wild type virus and challenge 

virus excretion was measured. Finally, the stability of the virus was 

determined by titration of the freeze-dried vaccine after storage at 

temperatures of +4˚C and -20˚C for more than one year. 

Results: There were no local or systemic reactions after vaccination 

observed in either of the two safety studies. With regard to efficacy, the 

serum-free produced vaccine performed equally well as the 

conventionally produced one and both met the strict requirements as 

laid down in the European Phamacopeia. Finally, the virus titers of the 

freeze-dried vaccines were stable for more than one year at 

temperatures of +4˚C and -20˚C. 

Conclusions: These data demonstrate the safety and efficacy of serumfree 

produced live vaccines in cattle as well as the stability of these 

products.

654 Complete Protection from Renal Infection with Leptospira 

Borgpetersenii Serovar Hardjo in Calves after Transfer of 

Specific Antibodies 

E. Klaasen 1 , T. Mols 1 , M. Smit 1 , B. Makoschey 2 

1 

Intervet-Schering-Plough, R&D, Boxmeer, The Netherlands 

2 



Objectives: The purpose of this study was to examine whether 

administration of serum antibodies can protect cattle from renal 

infection with Leptospira borgpetersenii serovar Hardjo, even if 

challenge is performed as long as 7 weeks after serum transfusion. 

Materials and Methods: Ten conventional 5-week-old calves free of 

serum antibodies against serovar Hardjo were randomly assigned to 

two groups of five animals each. Each calf of group 1 received a 

transfusion of a hyper-immune serum with a titer of 9.1 [log2/ml] of 

antibodies specific for serovar Hardjo. The calves of group 2 (control 

group) were left untreated. Seven weeks later all animals were 

challenged on three consecutive days by conjunctival instillation of 

Leptospira borgpetersenii serovar Hardjo genotype hardjobovis. 

Serum samples were taken from the calves of groups 1 and 2 

immediately prior to the transfusion as well as 24 hours, 4 weeks and 7 

weeks thereafter. Antibodies specific for serovar Hardjo, were 

measured by ELISA. Urine samples were taken pre-challenge and 2, 3, 

4, 5 and 6 weeks post-challenge to assess the effect of the challenge. 

The calves were sacrified 6 weeks post-challenge and kidney tissue was 

taken. Urine samples and kidney tissue homogenates were cultured. 

The cultures were examined every 3 weeks with dark-field microscopy. 

Cultures were considered negative when no leptospires were detected 

after 18 weeks of incubation. 

Results: The titration of serum samples taken before the transfusion 

confirmed that the calves of both groups were free of serum antibodies 

against serovar Hardjo and the untreated animals in group 2 remained 

seronegative until the time of challenge. By contrast, mean titers of 5.8, 

4.0, and < 3.3 were measured in the samples taken from animals of 

group 1 at 24 hours, 4 weeks and 7 weeks after transfusion respectively. 

Renal infection was demonstrated in four out of five control animals, 

whereas in none of the five pre-treated calves renal infection could be 

detected. 

Conclusion: In this study, complete protection of cattle from renal 

infection with serovar Hardjo was obtained with merely humoral 

immunity through passive immunization. In this study, the protection 

lasted for at least 7 weeks after transfusion. At this time point, the 

serum titre of antibodies had declined below the detection limit of the 

ELISA. 

655 Compatibility of a Live IBR Marker Vaccine and an 

Inactivated BVDV Vaccine 

B. Makoschey 1 , J. Munoz Bielsa 1 , L. Santos 2 , M. Alvarez 3 

1 



2 

Intervet-Schering-Plough, Laboratorios Intervet S.A., Salamanca, 

Spain 

3 

University of Leon, Animal Health Department, Leon, Spain 

Objectives: The target animals and vaccination regimes for vaccines 

against the bovine rhinotracheitis (IBR) and the bovine viral diarrhea 

virus (BVDV) are very similar. In order to simplify animal handling it 

would be desirable to combine the two vaccination procedures. 

Therefore, we have examined whether a live IBR marker vaccine 

(Bovilis ® IBR marker live, Intervet) and an inactivated BVD vaccine 

(Bovilis ® BVD, Intervet) can be applied at the same time. 

Materials and Methods: A total of 57 head of cattle were included in 

the study and seven groups of animals were established. The animals 

were vaccinated on days 0, 28 and 208 according to different 

(combined) vaccination schedules. The immune responses against the 

bovine herpesvirus type 1 (BoHV-1) and BVDV were determined and 

compared for the different vaccination schedules. 

Results: BoHV-1 antibody titers after simultaneous (together in the 

same syringe) or concurrent (two separate injections) application of the 

inactivated BVD vaccine were not lower than after vaccination with the 

BoHV-1 vaccine alone. To the contrary, BoHV-1 titers were even 

higher after simultaneous application at the first or second dose of the 

basic vaccination course of the BVD vaccine and at the booster 

vaccination. Likewise, BVDV antibody titers after simultaneous or 

concurrent application of the live BoHV-1 vaccine at the first or second 

dose of the basic vaccination course of the BVD vaccine and at the 

booster vaccination were not different from the titers after vaccination 

only with the BVD vaccine. 

Conclusions: There was no negative effect on the antibody response 

against either of the two vaccines when the live IBR marker vaccine 

was applied simultaneously (together in the same syringe) or 

concurrently (two separate injections) with the first or second dose of 

the basic vaccination course of the BVD vaccine and then at the booster 

vaccination (3rd dose). 

656 Compatibility of an Inactivated IBR Marker Vaccine and an 

Inactivated BVDV Vaccine 

B. Makoschey 1 , J. Donate 2 , L. Santos 2 , M. Alvarez 3 

1 



2 

Intervet-Schering-Plough, Laboratorios Intervet S.A., Salamanca, 

Spain 

3 

University of Leon, Animal Health Department, Leon, Spain 

Objectives: The target animals and vaccination regimes for infectious 

bovine rhinotracheitis (IBR) and the bovine viral diarrhoea virus (BVDV) 

vaccines are very similar. To simplify animal handling on the farm it 

would be desirable to combine both vaccination regimes. We have 

performed a study to assess the efficacy in terms of antibody response of 

the simultaneous (both vaccines in the same syringe) and concurrent (both 

vaccines at the same time, but different injection sites) use of Bovilis ® 

BVD (Intervet, The Netherlands) and Bovilis ® IBR marker inac (Intervet, 

The Netherlands) in dairy cattle under field conditions. 

Materials and Methods: The study was carried out as a randomised, 

partially blinded and controlled field trial on a commercial dairy farm 

in Spain. All animals included in the study were shown to be free of 

antibodies against the IBR and the BVD virus prior to the first 

vaccination. Forty animals were included in the study and assigned at 

random to one of the four groups of ten animals each. The vaccination 

protocols were as follows: group A received only inactivated IBR 

marker vaccine, group B received only BVD vaccine, group C were 

vaccinated simultaneously (in the same syringe) with both vaccines, 

group D were vaccinated concurrently (both vaccines at the same time, 

but in two separate injection sites) with the two vaccines. All animals 

were vaccinated twice (at admission (d0) and four weeks later (d 28). 

Results: The neutralizing antibody response against BVDV did not 

reveal any differences between the group vaccinated only with the 

BVD vaccine and the groups that were vaccinated simultaneously or 

concurrently with the IBR marker vaccine. Likewise, the neutralizing 

antibody titers against BoHV-1 did not exhibit any negative effect by 

the simultaneous or concurrent use of the two products as compared to 

the single IBR marker vaccination. 

Conclusions: These results indicate that the two vaccines can be 

applied at the same time either at the same or at different injection sites 

without any negative effect on the neutralising antibody response 

against the two vaccine viruses. 

657 A Serological Study on Bovine Viral Diarrhea - Mucosal 

Disease (BVD-MD) in Kurdistan Province 

Sh. Fakur 1 , Fa. Hemmatzadeh 2 

1 Faculty of Veterinary Medicine Islamic Azad University Sanandaj, 

Large Animal Clinical Sciences, Sanandaj, Iran 

2 Facutly of Veterinary Medicine, University of Tehran, Large Animal 

Clinical Sciences, Tehran, Iran 

Objectives of study: Bovine viral diarrhea disease is one of the viral 

diseases in cattle that was reported 10-90 percent in many countries. 

The causative RNA virus is a member of the family togaviridae and the 

genus pestivirus which is antigenically closely related with the virus 

of BD in sheep. Cattle are the most sensitive species to virus and are 

considered the principal reservoir of BVD viruses. A large proportion 

of BVD infections are subclinical and majority seropositive cattle. 

Since the virus has affinity for the lymphoreticular tissue this can result 

immonosuppression and abortion, produce calves with congenital 

abnormalities and persistant infectious (PI), So diagnostic laboratory 

procedures such as virus isolation and serologic methods are required. 

Materials and methods: In this study 410 serum samples were tested 

by serum neutralization test (SN) by using the NADL strain of BVD 

virus. 


Results: The cows sera were collected from Kurdistan province of 

Iran. By SN test the rate of infection in all samples was 27.7%. This 

result done with similar study was approximately like that one carried 

out on 1357 serum samples from other provinces in Iran. 

Conclusions: Being on the border and consequently animals entering 

the country explains the high rate infection in Kurdistan province. 

Key words: viral diarrhea, serum neutralization, Sanandaj 

658 Treatment of Infectious Diseases in Sheep with Enrofloxacin 

10% (Kinetomax) - Case Reports 

MI. Ferreira 1 , EJ. Facury Filho 2 , AU. Carvalho 2 , I. Borges 1 , 

G. Macedo Junior 1 , N. Martins 3 

1 

Escola de Veterinária da UFMG, Departamento de Zootecnia, Belo 

Horizonte, Brazil 

2 

Escola de Veterinária da UFMG, Departamento de Clinica e 

Cirurgia Veterinaria, Belo Horizonte, Brazil 

3 

Escola de Veterinária da UFMG, Departamento de Medicina 

Veterinária Preventiva, Belo Horizonte, Brazil 

Enrofloxacin 10% (Kinetomax) is a potent chemotherapist agent 

showing bactericidal action. It has a broad spectrum of antimicrobial 

action presenting rapid absorption, distribution and metabolism in the 

body. Thirty-two crossbred LacaunexSanta Ines sheep were allocated 

to individual stalls and kept in confinement in the Laboratory of 

Animal Calorimetry and Metabolism, School of Veterinary Medicine 

at UFMG, for 120 days in lactation for researching on production and 

composition of milk. Case 1: seven animals - five sheep and two lambs 

- had acute inflammatory reaction of the conjunctiva; sclera hyperemia; 

photophobia; blepharospasm; lacrimation; mucopurulent ocular 

discharges; and corneal opacity, without ulceration, resembling 

keratoconjunctivitis. Enrofloxacin 10% (Kinetomax) was used for 

treatment in accordance with the recommendation of 7,5 mg/Kg /b.w., 

in all affected animals. On the fourth day after application of the 

product, the reduction of clinical symptoms was observed, with 

complete recovery of the animals. Prior to the treatment onset, samples 

of ocular secretion were collected, with the help of sterile swabs, and 

sent to the Laboratory of Microbiology at the Department of Preventive 

Veterinary Medicine, where Staphylococcus caprae was isolated. Ten 

days after treatment, two animals showed clinical relapse and the 

occurrence of two new cases. The animals were subjected to treatment, 

following the fore-cited protocol. On the fourth day after the second 

treatment, there was complete reduction of clinical symptoms. Case 2: 

monitoring the daily consumption, a gradual decrease of food intake 

and reduced milk production of one animal was observed for five 

consecutive days. In the clinical examination, there was an increasing 

in volume, redness, and heat on the crown of the left forelimb hoof 

(coronary band), so that the animal could not stand, featuring an 

interdigital phlegmon (footrot), in initial state of development. For 

treatment, enrofloxacin 10% (Kinetomax) was used at a dose of 7,5 

mg/Kg /b.w., along with flumexine meglumine at the dose of 2,2 

mg/Kg. Hoof trimming was made in the hoofs where no deeper injuries 

were found. On the following days, there was an increase of food intake 

and the recovery of the clinical signs was complete on the third day 

after treatment. 

Key words: footrot, keratoconjunctivitis, Kinetomax, sheep 

659 Efficacy Following a Flexible Dose Regimen of Spirovac R , a 

Leptospira Borgpetersenii Serovar Hardjo (Type Hardjobovis) 

Bacterin, in a Challenge Study in Cattle 

D. Cleary 1 , S. Behan 1 , J. Wong 1 , M. Fisher 1 , S. Saginala 1 , L. Kopta 1 , 

M. Tory 1 , A. Weber 1 , G. Gallo 1 , R. Leyh 1 , J. Galvin 1 , J. Salt 2 

1 

Pfizer Animal Health, Veterinary Medicine Research and 

Development, Kalamazoo, Michigan, United States 

2 

Pfizer Animal Health, Animal Health Research Division, Sandwich, 

United Kingdom 

A research study was conducted to demonstrate the protection in cattle 

afforded by a flexible dosing regimen using a Leptospira 

borgpetersenii serovar Hardjo (type hardjo-bovis) bacterin (Spirovac, 

Pfizer Animal Health). All animal care and procedures were approved 

by an ethical review board for compliance with animal welfare 

guidelines. Spirovac (n=20) or placebo (n=11) treatments were 

randomly assigned to 4-week-old L. borgpetersenii seronegative 

[microscopic agglutination test (MAT) antibody titers of [1:12.5] heifer 

calves. The Spirovac’ group of calves received a second bacterin dose 


a minimum of four months later. Three weeks following administration 

of the booster dose, all animals, including the placebo-vaccinated 

control group, were challenged for three consecutive days via 

conjunctival instillation of a heterologous strain of L. borgpetersenii. 

Urine samples were collected for bacterial culture up to five weeks 

following challenge when the animals were humanely euthanized and 

kidney samples were collected. All animals were seronegative to L. 

borgpetersenii at the time of first vaccination and all placebo control 

animals were seronegative at the time of the challenge. The primary 

variable utilized for the evaluation of protection was bacterial isolation 

(presence or absence) of L. borgpetersenii in the urine following 

challenge and in the kidneys at necropsy. Ninety-one percent (10/11) of 

the placebo control animals were positive for L. borgpetersenii 

isolation in both urine and kidney samples, confirming a valid 

challenge. No L. borgpetersenii was isolated from the urine or kidneys 

in the group receiving two doses of bacterin (0/20). These results 

confirm that two doses of Spirovac administered approximately four 

months apart can protect calves against virulent challenge. 

Key words: Leptospira hardjo, Spirovac, vaccines, immunology, cattle 

660 Survey on Dairy Cows Infective Abortions in the North-East 

of Italy 

A. Barberio 1 , N. Pozzato 1 , L. Ceglie 2 , S. Nardelli 2 , M. Badan 1 , 

G. Vicenzoni 1 

1 

Istituto Zoop. Sper. Venezie, Sezione Verona e Vicenza, Vicenza, 

Italy 

2 

Istituto Zoop. Sper. Venezie, Dipartimento Sanita’ Animale, Legnaro 

(PD), Italy 

Objectives of study: The purpose of this study is to describe the results 

of the abortion surveillance program in Veneto region (North-East of 

Italy) over a period of two years. 

Materials and Methods: Aborted fetuses accompanied by the dam’s 

blood sample were delivered to the Regional Veterinary Laboratory 

(IZS), and submitted to a panel of laboratory tests. The cows sera were 

tested for antibodies against Brucella, Neospora, IBR, Leptospira, and 

BVD non structural protein NS2-3. ELISA tests were used for 

Neospora (VMRD), BVD (Pourquier) and IBR (Bommeli) antibodies 

detection, while microagglutination with the method of Martin and 

Petit was used for Leptospira. Brucella antibodies were detected 

following the EU regulation mandatory method. Necroscopy and 

routine bacteriology were performed on all fetuses, while Brucella spp 

isolation was carried out only if the abortion occurred after the 5th 

pregnancy month; Campylobacter fetus isolation was performed on 

abortions occurred before the 5th month. In case of positive results for 

BVD or N. caninum antibodies in mother serum, identification of these 

microorganisms in fetus was performed with RT-PCR for BVDV and 

PCR for Neospora. 

Results: During year 2005 and 2006, 577 abortions were delivered to 

IZS laboratories: 30% of abortions (170) were due to infective 

microorganisms and among these N. caninum was the most frequent 

(63%). Among the other microorganisms found, BVDV was isolated in 

14% of positive fetuses, Arcanobacterium pyogenes in 9%, and 

Listeria monocytogenes in 4%. Considering that only fetuses aborted 

by serological positive cows were tested with PCR for N. caninum, the 

percentage of fetuses found infected by this parasite was very high 

(47%), and is in agreement with the reports indicating that cows with N. 

caninum antibodies are more likely to abort than seronegative cows. 

Serological tests showed a high prevalence of antibodies against BVD 

virus (54%), Neospora caninum, (39%), and IBR virus (32%). On the 

contrary there was a low frequency (3%) of positive sera for 

Leptospira, and among the 16 positive samples, only two cows had a 

serological titer high enough to be considered the consequence of a 

recent infection. 

Conclusions: In our opinion the abortion surveillance program 

provided many useful and interesting information about the health 

status of dairy farms and the activities to be performed to prevent 

abortion diseases in the future, besides guaranteeing a mandatory 

requirement for the declaration of officially brucellosis-free region. 

661 Severe Outbreak of Malignant Catarrhal Fever in a Dairy 

Herd 

T. Autio 1 , P. Syrjälä 2 , M. Hautaniemi 3 , U. Rikula 3 , S. Pelkonen 2 

1 

Finnish Food Safety Authority Evira, Production Animal Health, 

Kuopio, Finland

2 

Finnish Food Safety Authority Evira, Veterinary Bacteriology, 

Kuopio, Finland 

3 

Finnish Food Safety Authority Evira, Veterinary Virology, Helsinki, 


Objectives of the study: Sheep-associated malignant catarrhal fever 

(SA-MCF) is a severe and mostly fatal disease of cattle and other 

ruminants. Severe outbreaks have been described in bisons that are the 

most susceptible animals. Sheep-associated MCF has been linked to 

ovine herpesvirus-2 (OHV-2) infection. We describe here an outbreak 

of SA-MCF with high mortality in a Finnish dairy farm. 

Materials and Methods: The farm. The affected herd consisted of 

appr. 40 dairy cows and heifers, calves and young cattle. All animals 

were housed in the same insulated barn with shared air conditioning, 

operated with forced exhaust ventilators. Free stall was used for cows, 

and young cattle were raised in group pens. The farmer brought two 

lambs, born in spring 2005, into the dairy barn in November 2005 and 

the lambs stayed in the barn until early April 2006. Examinations: 

Pathological examinations were carried out and diagnoses were 

verified by OHV-2 specific PCR. Serum and EDTA blood samples 

were collected from all living animals in early April 2006 for analyses 

of OHV-2 (PCR) and MCF-antibodies (c-ELISA). 

Results: In early January 2006 the exhaust ventilators were off for 

eight hours due to a power failure. Thus the air quality in the barn was 

extremely poor with high humidity and severe condensation. First signs 

of MCF were seen in February and the last case was in November 2006. 

The peak of the cases was at spring time. The sings including high 

fever, milk drop in cows, hyperaemic oral and nasal membranes, 

depression, oculo-nasal discharge and diarrhoea were seen in a total of 

31 animals including cows, calves, heifers and young cattle. One cow 

with typical clinical signs, high titre of specific antibodies and positive 

OHV-2 PCR result recovered. The remaining 30 animals with signs of 

MCF died or were euthanized. The diagnosis of MCF was based on 

gross and histopathology and presence of both OHV-2 DNA and 

specific antibodies. The animals were also tested for other agents of 

upper respiratory tract and intestinal infections including respiratory 

syncytial virus, infectious bovine rhinotracheitis, bovine coronavirus, 

bovine viral diarrhoeal virus, mycoplasmas and salmonella, with 

negative results. 

Conclusions: The adolescent sheep between 6 and 9 months of age 

have been shown to shed OHV-2 virus more often and in greater 

amounts than adults. Here keeping of two adolescent sheep in a dairy 

barn caused a severe outbreak of MCF in a dairy herd resulting in death 

of 30 animals. 

662 Evaluation of Nasopharyngeal Swabs in Sampling for 

Respiratory Pathogens in Calves 

T. Autio 1 , T. Pohjanvirta 2 , A. Huovilainen 3 , L. Sihvonen 3 , 

S. Pelkonen 2 

1 

Finnish Food Safety Authority Evira, Production Animal Health, 


2 

Finnish Food Safety Authority Evira, Veterinary Bacteriology, 


3 

Finnish Food Safety Authority Evira, Veterinary Virology, Helsinki, 


Objectives of the study: The aim of this study was to compare 

nasopharyngeal swab (NP) and bronchoalveolar lavage (BAL) samples 

in sampling for respiratory pathogens in calves. The samples were 

examined for presence of Pasteurella multocida, bovine coronavirus 

(BCV) and respiratory syncytial virus (RSV), which are the most 

common pathogens associated with calf respiratory disease in Finland. 

The similarity of Pasteurella multocida isolates from upper and lower 

respiratory tract was analysed. 

Materials and Methods: NP and BAL samples were taken in parallel 

from 42 calves. NP was taken with a guarded 27-cm swab and BAL 

with a guarded nasal tube. The samples were examined 

bacteriologically by standard laboratory methods. Pulsed-field gel 

electrophoresis (SalI) was utilized for genetic typing of P. multocida 

isolates. BCV and RSV were detected with RT-PCR. 

Results: Five animals were negative for the studied pathogens. Both P. 

multocida and BCV were detected in 14 animals. A combination of P. 

multocida, RSV and BCV was found in two animals, and three were 

infected with both P. multocida and RSV. A total of five animals 

harboured RSV, of which two in both samples and two only in NP. 

BCV was detected in nine animals by both sampling techniques and in 

eight and three animals only in NP or BAL, respectively. P. multocida 

was detected in 33 animals. Out of these, 23 calves harboured P. 

multocida both in NP and BAL. All BAL samples positive for P. 

multocida were also NP positive, and 10 only in NP. Paired P. 

multocida isolates obtained from 23 animals with both sampling 

methods were characterized by PFGE. Majority of animals (20/23) 

harboured isolates showing similar macrorestriction patterns (MRP) or 

had minor differences in their MRPs. 

Conclusions: Nasopharyngeal swabbing was found to be a suitable 

method for sampling for BCV, RSV and P. multocida. All pathogens 

were recovered more often in nasopharyngeal swab than in BAL 

samples. PFGE typing indicated that in most Pasteurella cases the same 

strain was isolated by both sampling methods. Thus the P. multocida 

culture obtained by nasopharyngeal swabbing represents reasonable 

well for the microbes in the lung. 

663 First Case of Immunohistochemical Detection of Mycoplasma 

bovis in Natural Cases of Calf Pneumonia in Hungary 

L. Szeredi 1 , T. Labossa 2 , M. Tenk 3 , V. Pálfi 1 , D. Rigó 1 , T. Molnár 1 

1 Central Agricultural Office, Budapest, Hungary 

2 Veterinary Practitioner, Lajoskomárom, Hungary 

3 CEVA, Budapest, Hungary 

Objectives of the study: Bovine respiratory disease complex (BRDC) 

is a major cause of morbidity, mortality and economic losses in cattle 

production systems. The cause of this disease is rather complex 

involving both managing factors and infectious agents (bacteria and 

viruses). In our study, we demonstrate the primary role of Mycoplasma 

(M.) bovis in a severe respiratory disease in feedlot cattle. 

Materials and methods: Five feedlot cattle died after a few days of 

illness without any specific symptom in a herd. Out of these 2 were 

treated with antibiotics whereas 3 suddenly died without any treatment. 

All the animals were imported from Poland, vaccinated previously 

against infectious bovine rhinotracheitis (IBR) and bovine respiratory 

syntitial virus (BRSV). After gross pathological investigation swabs 

were collected for bacterial culture. From the lungs, mycoplasma 

culture was also made in 1 case and virus isolation (VI) was attempted 

in 4 cases. Tissue samples were collected from the lungs and 

occasionally also from the trachea and the mediastinal lymph nodes for 

histological examination. Immunohistochemical (IHC) method was 

used to detect viruses (bovine herpesvirus 1 (BHV-1), BRSV, bovine 

respiratory coronavirus, influenza A virus) and bacteria (M. bovis and 

Chlamydiaceae) in formalin-fixed paraffin-embedded tissue samples. 

Results: The cranioventral part or the whole lungs were affected by 

dark discoloration in all cases. Histologically, fibrinopurulent 

pleuropneumonia, suppurative bronchiolitis and focal to coalescing 

coagulative necrosis were observed in the lungs in all cases. M. bovis 

was detected in large numbers in the lung tissue samples with IHC, 

mostly in the area of bronchiolitis and coagulative necrosis, and in 

smaller number in the area of the fibrinopurulent inflammation. M. 

bovis was isolated from one of the cases. Other pathogens were 

detected only infrequently: parainfluenza 3 virus (1 case with VI), RSV 

(1 case with IHC), Pasteurella multocida (2 cases with culture), 

Mannheimia haemolytica (1 case with culture). 

Conclusions: M. bovis was the most important pathogenic agent that 

induced a severe respiratory disease in the feedlot cattle examined in 

this study. Other bacteria and viruses were detected only in a few cases. 

In spite of the sudden death in 3 cases, the lung lesions were not acute 

in any of the cases. 

664 Short-duration Antimicrobial Treatment of Bovine 

Respiratory Disease with Enrofloxacin, Florfenicol and 

Cefquinome 

R. Froyman 1 , C. Boda 2 , C. Rizet 3 , A. Valognes 4 , N. Brunner 1 , 

P. Liege 2 , H. Navetat 3 

1 Bayer HealthCare, Animal Health Division, 51368 Leverkusen, 

Germany 

2 Anisteme Biosciences, Animal Health Division, 34400 Saint- 

Christol, France 

3 Centre d’Information Veterinaire Clinique, 03130 Le Donjon, 

France 

4 LDV03, 03017 Moulins, France 

Mannheimia haemolytica (Mh) is the most prevalent causative 

bacterium in bovine respiratory disease (BRD) but Mycoplasma bovis 


(Mb) as co-infecting agent is becoming increasingly important. In spite 

of this, the possible involvement of Mb is mostly overlooked whilst 

choosing the appropriate antimicrobial treatment of BRD-affected 

cattle. This trial, applying short-duration treatments, compared 

compounds (florfenicol [FFC], cefquinome [CFQ]) that are specifically 

directed against Mh, with enrofloxacin (ENR) that targets both Mh and 

Mb. Following Mb and subsequent Mh challenge, performed 4 days 

apart, groups of young calves (n=10/group) were treated 3 h later with 

ENR (Baytril Max 10%, 7.5 mg/kg, one SC injection), FFC (Nuflor 

30%, 20 mg/kg, 2 IM injections 48 h apart) or CFQ (Cobactan LA 

7.5%, 2.5 mg/kg, 2 SC injections 48 h apart). All treated calves 

survived the study but 2/10 untreated control calves died on day (d) 5 

after Mh challenge. Whereas the surviving untreated controls continued 

to have fever or high body temperature until the end of the study (d7 

after Mh challenge), treatment with ENR and FFC controlled fever 

within 15 h. It took 72 h to bring down body temperature to baseline 

level in calves treated with CFQ. None of the calves treated with ENR, 

FFC or CFQ showed respiratory disease signs or morbidity compared 

with 4 control calves in which such symptoms persisted until the study 

end. At necropsy pneumonia lesions were seen in all control calves and 

in 2/10, 4/10 and 8/10 calves treated with ENR, FFC and CFQ, resp. 

The median pulmonary lesion scores (scale 0-4) were 1.56, 0, 0, and 

0.44 in untreated, ENR, FFC and CFQ groups, resp. Mean Mh counts 

in lung tissue of ENR and FFC treated calves were very low (

2 University of Parma, Department of Animal Health, Parma, Italy 


Objectives: The control of the Bovine Viral Diarrhea Virus infection 

includes the identification and removal of persistently infected (PI) 

animals and/or vaccination. Some inactivated vaccines, as Bovilis ® 

BVD (Intervet International), have proven foetal protection, preventing 

the birth of new PI animals. This study had two objectives: (i) to 

monitor transplacental infection and (ii) to assess the efficacy of 

vaccination under field conditions. 

Materials and Methods: Six farms with a total of 1332 animals were 

included in the study. Three of them were not vaccinated, one was 

previously vaccinated with a modified live ts BVD vaccine and two 

with an inactivated BVD vaccine applied in a vaccination program for 

which foetal protection was not demonstrated. At the beginning of the 

study 76.5% of the animals were positive for BVDV NS2-3 antibodies. 

For the identification of PI animals and newborn calves two different 

methods were used: an antigen ELISA and PCR. All animals included 

in the study were vaccinated with a commercial inactivated BVD 

vaccine (Bovilis BVD) administered as two doses, 28 days apart 

(priming) and a booster every 6 months. The monitoring for detection 

of PI in newborns continued for 1 to 3 years. 

Results: At the beginning of the study, a total of 19 PI animals, was 

detected in 4 of the 6 farms. In one of these farms the PI animals were not 

immediately removed and remained in close contact with the other 

animals for about 10 months. The monitoring of the newborns showed 

that one new PI animal (0.12% of the 831 tested) was identified during the 

study. The mother of this calf, had been tested negative at the beginning of 

the study with ELISA antigen test, but was later on identified as PI when 

retested by PCR after the birth of her PI calf. This newborn PI can 

therefore not be related to a vaccine failure. Moreover, the PI mother was 

for at least 9 months in close contact with the other animals. 

Conclusions: The combined approach based on the identification and 

removal of PI and vaccination is an effective method of BVD infection 

control and the vaccination with the inactivated vaccine used in the 

study induced a high level of foetal protection even in presence of PI 

animals. 

669 Influence of BVDV Vaccination on Reproductive 

Performances in Field Conditions 

G. Valla 1 , F. Cammi 2 , J. Munoz Bielsa 3 , B. Makoschey 3 , F. Petrera 2 , 

M. Bussacchini 1 

1 

Intervet-Schering Plough, Intervet Italy, Peschiera Borromeo, Italy 

2 

Azienda Sperimentale Vittorio Tadini, Intervet Italy, Gariga di 

Podenzano (PC), Italy 

3 

Intervet-Schering Plough, Int. Marketing, Boxmeer, The Netherlands 

Objectives: BVDV infection can cause transplacental infection, heat 

returns and abortions. When the infection occurs during oestrus it can 

induce ovarian lesions and hormonal disorders, and therefore reducing 

the pregnancy rate. A study was set up to evaluate the benefits of 

vaccination with a commercial inactivated BVD vaccine (Bovilis 

BVD, Intervet) on the reproductive performance. 

Materials and Methods: The trial was conducted in a dairy farm of 

about 120 lactating cows (Azienda Sperimentale Vittorio Tadini). A 

number of parameters, including days open (DO value), Average 

Service Per Pregnancy (ASPP), Heat Detection Rate for Repeat 

Insemination (POSTHDR), Conception Rate (CR) and Pregnancy Rate 

(PR), were recorded during 9 years (1998-2006) using a specific 

software (Fertility Factor Farmer, Italy). Three different vaccination 

regimens were implemented during that period. Between May 1998 

and April 2001 (Period A) all the animals were vaccinated, every 6 

months, with an IBR marker modified live vaccine; in period B (May 

2001 - April 2003), the farm was unvaccinated. From May 2003 to 

April 2006 (C) all the animals were vaccinated concurrently, every six 

months, with a modified live IBR marker vaccine and an inactivated 

BVD vaccine. Before starting BVD vaccination in April 2003, the herd 

was tested to identify PI animals. 

Results: Three PI were detected but not immediately removed. They 

remained in close contact with the other animals for about ten months. 

Despite the fact that no new PI animals were detected between 2003 

and 2006, BVDV circulation was noticed through the seroconversion 

against NS2-3 antibodies. During the same period the IBR percentage 

of IBR gE positive animals decreased from 23% to 8%. The DO value 

was 152 days in period A, 154 in B and 146 in period C. The ASPP 

decreased from 1.87 in A and 1.55 in B to 1.52 in C. The POSTHDR 

increased from 40.0% in A and 37.4% in B to 45.1% in C. The CR and 

the PR were 46.1% and 14.3 in A, 51.3% and 13.3 in B and 53.1% and 

14.8 in C, respectively. The profitability index, calculated for a 

lactation of 305 days and an average milk production of 9200 litres, 

was higher in the period C showing a difference in the revenue of 8 

/cow/year versus period A, and 25 versus the unvaccinated period B. 

Conclusion: The introduction of BVD vaccination improved 

reproductive such as DO value and POSTHDR, resulting in an 

economical benefit for the herd. 

670 Frequency of Isolation and Combitation of Pathogens 

Involved in Bovine Respiratory Disease in France: a 

Retrospective Analysis 

F. Payot 1 , E. LeDrean 2 , M. Bonnier 2 , B. Makoschey 3 , C. Dore 2 

1 

Intervet-Schering Plough, Farm and Production Animals Business 


1 

LDA35, Rennes, France 

3 


Objectives: Bovine respiratory disease (BRD) is generally recognized 

as a multifactorial disease. Primary causative pathogens, such as the 

bovine respiratory syncytial virus (BRSV), and secondary pathogens 

(enhancing the impact of primary pathogens), such as bovine 

parainfluenza type virus (PI3), are commonly discerned. We have 

studied the frequency of isolation of pathogens from calves with BRD 

and the frequency of combination of these pathogens (i.e. found alone 

or combined). 

Material and Methods: The analysis was performed on data from the 

Departmental Veterinary Laboratory 35 (Rennes, France) from 

November 2006 to October 2007. The samples originated from 

recently infected animals with BRD, mostly coming from the western 

part of France. The presence of the following pathogens was 

investigated: bacteria: Pasteurellaceae (Mannheimia haemolytica, 

Pasteurella multocida and Histophilus somni), Arcanobacter pyogenes 

and Mycoplasma bovis, viruses: Bovine viral diarrhoea virus (BVDV), 

BRSV, bovine herpesvirus type 1 (BoHV-1) and PI3. Only samples 

that were tested for all these pathogens were included in this 

retrospective analysis. 

Results: 180 samples were included. They were obtained from 

respectively 65%, 8% and 27% of transtracheal lavage, nasal swabs 

and lung tissue at necropsy. 52 samples were found negative and 128 

were positive for at least one pathogen. A combination of two or more 

pathogens was found in 56% (72) of cases whereas the presence of a 

single pathogen was observed in 44% (56) of cases. Globally, 

Pasteurellaceae, BRSV, M. bovis were isolated respectively from 

73%, 35% and 47% of samples. The other pathogens were detected in 

less than 15% of samples. Focused on single isolated pathogens (n=56), 

the following prevalences were observed: Pasteurellaceae 50%, BRSV 

25%, M. bovis 17%, A. pyogenes 4%, PI3V 4%. The frequency of 

combination of Pasteurellaceae or BRSV with other pathogens was 

67%-70%. In contrast, the frequency of combination of PI3V, M. bovis 

or A. pyogenes with other pathogens was higher: 83% for each. BVDV 

and BoHV-1 were always combined with other pathogens. 

Conclusion: This retrospective analysis emphasized the multifactorial 

aetiology of BRD. Pasteurellaceae and BRSV were frequently isolated 

combined or as single pathogens. M. bovis were also frequently 

isolated but mostly associated with other pathogens as observed for A. 

pyogenes and PI3. 

671 Comparison of Different Vaccination Protocols against 

Bovine Respiratory Disease Complex in Fattening Units 

C. Tresse 1 , K. Risson 1 , F. Bendailh 2 , B. Makoschey 3 , L. Oliviero 4 

1 

Ter elevage, Villedieu la Blouere, France 

2 

Ecole Vétérinaire, Toulouse, France 

3 


4 

Intervet-Schering Plough, Farm and Production Animals Business 


Objectives: Respiratory disease outbreaks are common after mixing and 

transportation of calves. In a field trial the efficacy of vaccination of calves 

in the breeding farm prior to transport was compared to vaccination after 

transport and mixing with animals from different origins. 

Materials and Methods: In the study, 161 calves at the age of 7 to 8 

months were included. They originated from 15 farms and were first 

sent to 2 trading centers for approximately 12 hours and from there 


distributed over 13 different fattening farms. In the farms of origin, the 

calves of group EV (early vaccination) were either vaccinated twice at 

1 month interval with a trivalent vaccine (bovine respiratory syncytial 

virus (BRSV), bovine parainfluenza type 1 (PI3), Mannheimia 

haemolytica), or twice at 1 month interval with a bivalent vaccine 

(BRSV, bovine viral diarrhoea virus (BVDV)). The vaccination of 

group EV was completed 15 days before transport to the trading center. 

A second group LV (late vaccination), of calves, that arrived at the 

trading centers at the same time as the EV group, was left unvaccinated 

until the arrival at the trading center. The animals of the group LV were 

vaccinated with the bivalent vaccine at the trading center (first dose) 

and the fattening farm (second dose, 1 month later). The two groups 

were raised under the same conditions in each fattening unit. The 

following parameters were recorded: morbidity and mortality due to 

respiratory disease, number of treatments due to respiratory disease 

during the first two months at the fattening unit. The observations were 

performed by persons that were not aware of the grouping. 

Results: The results showed a significant decrease in morbidity (5% 

versus 12.7%) and number of individual treatments (5.4% versus 18%) 

for the animals that received their vaccination at the breeding farm 

(group EV) as compared to the calves that were vaccinated at the 

trading center and the fattening unit (group LV). 

Conclusions: Timing of vaccination has a significant effect on the 

prevention of respiratory disease under the conditions described above. 

672 The Use of ELISA for the Diagnosis of Goat Tuberculosis in 

Brazil 

C. Marassi 1 , C. Almeida 2 , S. Pinheiro 2 S. Vasconcellos 2 , 

W. Lilenbaum 1 

1 

Universidade Federal Fluminense, Microbiology, Niterói - RJ - 

Brasil, Brazil 

2 

Universidade de Sao Paulo, Preventive Medicine and Public Health, 

Sao Paulo, Brazil 

Bovine tuberculosis remains an endemic infection in several countries, 

despite several efforts to control the disease. In the past, it was believed 

that goats were naturally resistant to tuberculosis; however it is now well 

established that M. bovis and M. caprae could be responsible for this 

disease in goats. After one clinical case that was presented to the 

veterinary service of the University of Sao Paulo, Brazil, a complete 

screening by intradermal tuberculin test was performed in the herd. The 

goat herd was composed by 500 animals and 83 (16.6%) of them showed 

to be reactive to the comparative double cervical intradermal test. Four 

months after the test, all the reactive animals were slaughtered and 33 of 

them were randomly chosen for histopathological, serological and 

bacteriological procedures. Histopathology (Ziehl-Neelsen and 

Hematoxyllin-eosin) evidenced at least one characteristic lesion of 

tuberculosis in each animal, with typical granulommas where BAAR 

could be observed. Bacteriology (inoculation in Stonebrink-Lesslie 

media) was positive for M. bovis in 25 samples (75.8%), therefore 

confirming the etiology of the outbreak. Sera of these 33 animals were 

tested in an ELISA using the recombinant M. bovis protein MPB70 as 

capture antigens. From those, 31 were reactive to the test, with high ODs 

results, considering a cut-off point established by ROC curve analyzing 

results of M. bovis culture from tuberculous cattle (cut-off = 0.06; mean= 

0.55; range: 0.157-1.357). These preliminary results suggested that 

MPB70-ELISA can be considered as a reliable tool to diagnose 

tuberculosis in goat herds, since this assay was capable to demonstrate the 

disease in 93.9% of the animals here examined. 

Key words: tuberculosis, goats, ELISA, diagnosis, M. bovis 

673 Surveillance of Bluetongue Virus in Domestic Ruminants in 

Territory of the Slovakia- Draft of Research Program 

J. Bires, A. Kocisova, M. Mojzis, J. Pliesovsky, V. Petrovic 

University of Veterinary Medicine, Department of Cattle and Sheep 

Diseases, Kosice, Slovakia 

The research project is focused on acquirement of primary information 

about occurrence of health and economically serious orbivirus 

transmissive disease - bluetongue in cattle, sheep and goats in Slovakia. 

In content of project there will be the research of occurrence of vector 

Culicoides spp. and their position towards transmission of catarrhal 

fever on animals (entomological study), monitoring of potential 

occurrence of disease in cattle, sheep and goats (clinical and laboratory 

examinations) and verification and introduction virological diagnosis 


of aetiological factor of bluetongue. Diagnostic procedures of 

bluetongue on the level of orbivirus, vector and animals are in 

accordance with Manual of diagnostic tests and vaccines for terrestrial 

animals (OIE) and legislative issues of European Commission. The 

intensity of surveillance bluetongue will be realised on selected areas 

of Slovakia during whole year on all the levels of observation 

depending on climatic conditions and persistency of vectors in nature. 

Solution of project expects collaboration of experts from veterinary 

university, state veterinary institute, state veterinary administration, 

veterinary institute, veterinary surgeon and farmers. 

Key words: bluetongue, surveillance, domestic ruminants 

674 Prevalence of BVDV Infection in Large and Small Cattle 

Herds in Southern Backa and SREM District in Serbia 

T. Petrovic 1 , S. Lazic 1 , D. Bugarski 2 , B. Djuricic 3 

1 Scientific Veterinary Institute Novi Sad, Virology, Novi Sad, Serbia 

2 Scientific Veterinary Institute Novi Sad, Department of 

Epizootiology and Health Protection of Ruminants, Novi Sad, Serbia 

3 Faculty of Veterinary Medicine Belgrade, Department for Infectious 

Diseases- Epidemiology, Novi Sad, Serbia 

In order to determine the prevalence of BVDV infection in small and 

large cattle herds in Southern Backa (S. Backa) and Srem district, the 

examination was carried out on presence of VN antibodies in blood 

sera in breeding cattle that were older than 6 months during 2004. The 

examined 3457 samples were from small cattle herds in S. Ba_ka (9 

communities) and 4120 samples in Srem district (8 communities). The 

examination included animals from all the settlements (156) in this 

area. The examined 2721 samples were from 18 large cattle herds from 

S. Backa, and 298 samples were from 4 large cattle herds from Srem 

district. The examination included animals from all large herds (22) in 

this area. Virus neutralization test (VN) was performed according to 

standard procedure with small modifications. In the test NADL strain 

of BVD virus and cell culture MDBK were used. The sera were 

examined in double solution from 1:2 to 1:512.The presence of VN 

antibodies was discovered in 1833 (24.19%) samples from small cattle 

herds. Out of this number in S. Backa district there were 1082 

(31.30%), and in Srem district there were 751 (18.23%) seropositive 

animals. The largest number of seropositive animals in S. Backa 

district was determined in Titel municipality (68.07%) and the smallest 

in Backi Petrovac municipality (6.63%). In Srem district the highest 

percent of seropositive animals was found in Beocin municipality 

(34.16%) and the lowest in Irig municipality (7.62%). The presence of 

VN antibodies was discovered in 1667 (55.22%) samples from large 

cattle herds. Out of this number in S. Backa district large herds there 

were 1500 (55.13%), and in Srem district large herds there were 167 

(56.04%) seropositive animals. The prevalence of seropositive animals 

in big cattle herds from S. Backa district ranged from 0% (5 large 

herds) to 96.61%. High prevalence (>70%) was determined in 7 out of 

18 herds. The prevalence of seropositive animals in large cattle herds 

from Srem district ranged from 0% (1 big herd) to 96.96%. High 

prevalence (>70%) was determined in 2 out of 4 herds.The obtained 

results point out that BVD infection is spread on the examined area. At 

the same time, prevalence is not equal, what means that there are areas 

where it is not present, or is only sporadically present. 

Key words: BVDV, VN test, prevalence 

675 Antimicrobial Resistance in Escherichia coli Isolated from 

Calves in Slovakia 

V. Kmet 1 , D. Bujnakova 1 , D. Demankova 1 , S. Novak 1 , R. Bobcek 2 , 

T. Szakal 2 , M. Huska 2 

1 

Slovak Academy of Sciences, Institute of Animal Physiology, Kosice, 

Slovakia 

2 



A total of 159 faecal Escherichia coli isolates obtained from calves 

from seven regions in Slovakia during one year were examined for their 

quantitative susceptibility. MIC values to 20 antimicrobial agents were 

determined by a colorimetric microdilution test. The underlying 

resistance mechanism were tentatively identified from antibiogram 

profiles using interpretative reading method. The most frequently 

found resistance was to tetracycline (76 %), streptomycin (77 %) and 

ampicillin (78 %). Among tested E . coli isolates also high resitance 

rates to neomycin (51 %), chloramphenicol (30 %), cotrimoxazole (28

%) and spectinomycine (25%) could be recorded. Resistance to 

ampicillin+sulbactam (8,1 %), ceftiofur (4,4%), apramycin (0.6 %), 

gentamicin (7 %) and florfenicol (10 %) was lower. All of the tested E. 

coli isolates were sensitive to cefquinome and colistin. Resistance to 

ciprofloxacin (17 %) and enrofloxacin (16,9%) was intermediate. 

However, overall quinolone resistance detected by means of nalidix 

acid was as high as 29 %. The high level fluoroquinolone resistance 

(CIP: 4mg/L and ENR: 16 mg/L) (18%) as well as penicillinases 

((TEM1,2/SHV1)-low (51%) and (TEM1,2/SHV1)-high (25%) was 

detected in E. coli isolates. This is the first report of a quantitative 

antibiotic susceptibility testing and phenotype resistance mechanisms 

determination in E. coli isolated from calves in Slovakia. 

Key words: E. coli, minimal inhibitory concentration, interpretative 

reading, quinolone resistance, calves 

676 Deciphering the Value of Chlamydia-Serology in Cattle 

J. Boettcher, A. Vossen, A. Gangl, N. Meier, G. Wittkowski 

Bavarian Animal Health Service, Poing, Germany 

Chlamydiae are often suspected in reproductive and respiratory diseases 

of cattle. Because of frequently used ELISA-testing, we tried to assess its 

value. Based on 21683 sera a seroprevalence (SerP%) of 37.0%±0.3% 

was determined for Bavaria (Idexx-ELISA). Positive sera were detected 

in 92.8%±0.9% of the farms (n=765). Average within-herd prevalence 

was 37.0%±1.4%. Age-related effects (n=3522): SerP% was highest in 

calves less than 9 month of age (mona) (48%±6%) and lowest between 

21-24 mona (41%±2%)(n.s.). On 5 clinically unaffected farms swabs 

were analysed for Chlamydia by PCR (357 animals). Detection rates for 

vaginal, nasal and conjunctival swabs were 2.5%±0.8%, 2.0%+0.7% and 

1.1%±0.6%, respectively. Detection rate was 22% for calves 12 months (p=0.014). SerP% in ELISA were 36% and 10% for Idexx 

and Pourquier, respectively. While SerP% (Idexx) increased from 22% in 

calves

main infection sources to the urban rabies, while the chiropters are 

important in rabies transmission to the production animals. Due to 

the relevance of this zoonosis to Public Health some measures of 

control are usually implemented: vaccination of dogs and cats, 

laboratorial diagnosis surveys, control of the chiropters, sanitary 

educational programs. The purpose of this study was to refer the 

positive rabies diagnosis from 2006 to 2007, conducted by the 

Zoonosis Diagnostic Center- FMVZ/UNESP-Botucatu/SP-Brazil 

staff. A total of 1061 brain samples from various animal species 

were analyzed during this period, from different counties sent by the 

epidemiological surveillance groups, GVEs 15 and 16. Direct 

immunofluorescence technique and the biological test were 

performed to rabies diagnosis. During the studied period 13 (1.23%) 

were positive, from these positives samples 38.46% (5/13) were 

from bovines and 61.54% (8/13) were from chiropter. When the total 

of bovine and chiropters samples was considered it was verified that 

the rabies positivity among the bovines were higher than among 

chiropters and the difference was statistically significant (P = 

0.0032, Fisher’s Test, confidence interval of 95%). In the same 

period and region it was verified negative results on 300 dog and 95 

cat samples. Recently, January, 2008, an outbreak occurred in two 

cattle farms. In one of them, 10 percent of 340 bovines showed signs 

of chiropters bites, five presented rabies symptoms and died. In the 

other, 10 bovines from a total of 149, showed chiropters bites signs, 

six of them presented the classical rabies symptoms and died. In both 

herds anti-rabies vaccination was not used. The rabies diagnosis 

from these animals was confirmed by the laboratory tests. The 

results of this study demonstrated that the rural rabies transmitted by 

haematophagus chiropters still being a problem in Brazil. As a 

conclusion, it can be inferred that in spite of the adopted measures to 

control the urban rabies were effective in the studied region the 

problem of rural rabies was still prevalent. It was closely linked to 

the chiropters as rabies Lyssavirus reservoir, therefore, they 

represent a real risk to the persistence of the rabies in bovines. 

Key words: rabies, zoonosis, public health, chiropter, bovine 

680 Prevalence and Risk Factors for Campylobacter sp. 

Colonization in Cattle in Poland 

M. Bednarski, A. Wieliczko 

Wroclaw University of Environmental and Life Sciences, Department 

of Epizootiology and Veterinary Administration with Clinic of 

Infectious Disease, Wroclaw, Poland 

In developed countries campylobacteriosis is now one of the most 

frequent reasons of foodborne diseases and usually it is associated with 

gastroenteritis. Animals and animal food products are the main source 

of the infection of Campylobacter for human. Because of the potential 

linkage between Campylobacter spp. harbored by cattle and human 

disease, in this study we investigated the prevalence and risk factors for 

colonization Campylobacter spp. in cattle. 

Objectives of study: The objectives of the study were to estimate 

prevalence and risk factors for colonization Campylobacter spp. in 

cattle in Poland. 

Materials and Methods: The study was performed on 896 cows from 

84 farms located in South-Western Poland. Fecal samples were 

checked in two seasons: summer and winter. All fecal samples were 

analyzed by microbiological and PCR methods. Isolated 

Campylobacter sp. strains were characterized by fenotypic and 

genotypic methods. Prevalence data analysis was performed using 

Pearson chi-square test. 

Results: From all fecal samples tested by microbiological method 151 

(16,85%) were positive for Campylobacter sp. C. jejuni was the most 

commonly isolated species with 143 positive fecal samples (94,7% of 

all isolated Campylobacter strains) whereas C. coli were present only in 

7 samples (4,6% of isolated Campylobacter strains). Using PCR method 

29,4% of samples were Campylobacter positive. Campylobacter spp. 

prevalence in cattle were correlated with: extensive type of herds, 

outdoor and grazing cattle, cows grouped by-products, type of cows 

housing and husbandry, TMR (total mixed ration), not chlorinated 

drinking water, presence of poultry and wild birds on farm. 

Conclusions: The study showed relatively high prevalence of 

Campylobacter spp. in cows. Therefore, Campylobacter spp. have to 

be regarded as an important hazard of zoonotic disease in cow farms. In 

most cases this pathogen was isolated in fecal samples taken from 

clinically normal animals. 

Key words: Campylobacter, foodborne diseases, risk factors 


681 Antimicrobial Resistance of Thermophilic Campylobacter 

Spp. from Cattle Farms in Poland 

M. Bednarski, A. Wieliczko 

Wroclaw University of Environmental and Life Sciences, Department 

of Epizootiology and Veterinary Administration with Clinic of 

Infectious Disease, Wroclaw, Poland 

Campylobacter species are among the most frequently identified 

bacterial causes of human gastroenteritis. Because Campylobacter spp. 

harbored by cattle can be transmitted to human, in this study we 

investigated antimicrobial resistance of thermophilic Campylobacter 

isolated from cows. 

Objectives of study: The objectives of the study were to estimate 

antibiotic resistance of thermophilic Campylobacter isolated from 

cows in Poland. 

Materials and Methods: Our study included 150 strains of 

thermophilic Campylobacter (143 strains of C. jejuni and 7 strains of C. 

coli) collected from cows in South-Western Poland. Isolated strains of 

Campylobacter sp. were characterized by fenotypic and genotypic 

methods. The MICs to ciprofloxacin, erythromycin, gentamicin, and 

tetracycline hydrochloride were determined using the agar dilution 

methodology according to the CLSI. C. jejuni ATCC 33560 and C. coli 

ATCC 33559 were used as a control in susceptibility testing and also as 

a growth control strains. The MIC breakpoint used for the resistance to 

antibiotics was recommended by the National Committee for Clinical 

Laboratory Standards (NCCLS) for non-Enterobacteriaceae. 

Results: All strains of C. coli were susceptible to all four of the 

antimicrobial drugs studied. The most frequently detected resistance of 

C. jejuni was to ciprofloxacin (26 strains 18,2%). Resistance to 

tetracycline was observed in 5 (3,5%). All strains of C. jejuni were 

susceptible to erythromycin and gentamicin. 

Conclusions: Ciprofloxacin, erythromycin, gentamicin, and tetracycline 

were chosen for antimicrobial susceptibility testing in this study, because 

of their importance as a front-line therapeutic drug in humans and animals 

and the relatively frequent occurrence of resistance to these drugs. In 

conclusion, we demonstrated relatively high resistance to ciprofloxacin of 

C. jejuni. These data support the concept of continuous selection pressure 

for the emergence and spread of fluoroquinolone resistance in many parts 

of the world and in Poland as well. 

Key words: antibiotic resistance, Campylobacter, foodborne diseases 

682 Frequencies of PRNP Gene Polymorphisms in Cattle in 

Vietnam and Thailand for Potential Association with BSE 

Y. Muramatsu 1 , M. Horiuchi 2 , T. Ogawa 3 , K. Suzuki 4 , 

M. Kanameda 5 , T. Hanh 6 , C. Tongchai 7 , I. Kuramontong 7 , 

P. Kriengsak 7 , Y. Tamura 1 

1 Rakuno Gakuen University, School of Veterinary Medicine, Ebetsu, 

Japan 

2 Hokkaido University, School of Veterinary Medicine, Sapporo, 

Japan 

3 Nippon Veterinary and Life Science University, Faculty of 

Veterinary Science, Musashino, Japan 

4 Universidad National de La Plata, Facultad Ciencias Veterinarias, 

La Plata, Argentina 

5 Japan International Cooperation Agency, Tokyo, Japan 

6 National Institute of Veterinary Research, Department of Veterinary 

Hygiene, Hanoi, Vietnam 

7 Chulalongkorn University, Department of Veterinary Microbiology, 

Bangkok, Thailand 

Objectives of study: Since 2004, significant associations between 

bovine spongiform encephalopathy (BSE) susceptibility in cattle and 

frequencies of insertion/deletion (ins/del; indel) polymorphisms within 

the bovine prion protein gene (PRNP) have been reported. To know the 

genetic background in the PRNP gene in cattle in Asian countries, we 

investigated the frequencies of indel polymorphisms within two 

variable sites, a 23-bp indel polymorphism in the promoter region 

(23indel) and a 12-bp indel polymorphism in intron 1 region (12indel), 

in the bovine PRNP in cattle in Vietnam and Thailand. 

Materials and Methods: A genomic DNA was extracted from 278 

serum samples of Vietnamese cattle or 113 spleen samples of Thai cattle 

with a commercial kit in accordance with the manufacturer’s instructions 

(QIAamp DNA Mini Kit, QIAGEN). The extracted DNA was subjected 

to PCR methods for detection of 23indel and 12indel polymorphisms in 

the bovine PRNP. Both PCR methods were performed as previously

described with slight modification (Sander et al., 2004). Briefly, a total of 

50µl volume of reaction mixture containing 10 to 20µl volume of the 

extracted DNA was used for the first stages of both PCR methods. In case 

that no amplified DNA fragment was detected, 10µl each volume of the 

first PCR product was used as a template of the second PCR. The second 

PCR was performed in the same manner as the first PCR. 

Results: We could detect polymorphic DNA fragments in both of the 

23indel site and the 12indel site of 206 Vietnamese cattle and 102 Thai 

cattle. As for the 23indel in Vietnamese cattle, the frequency 

distributions of the del polymorphisms, which are thought to be 

associated with BSE susceptibility, were significantly high. However, 

no significant difference was shown in the frequencies of both allelic 

and genotypic polymorphisms in the 23indel site of Thai cattle. On the 

other hand, the frequencies of del allelic and del/del genotypic 

polymorphisms in the 12indel site, which have been reported to confer 

BSE susceptibility, were significantly low in cattle of both countries. In 

particular, the frequencies of del polymorphisms in the 12indel site 

were extremely low in Thai cattle. 

Conclusion: We have provided evidence that cattle in Vietnam and 

Thailand have a unique genetic background in the PRNP gene in 

comparison with cattle or sires previously reported in other countries. 

Key words: bovine PRNP, BSE, Vietnam, Thailand, indel 

polymorphisms 

683 Bluetongue Disease in Cattle in Northrhine-Westfalia Clinical 

Symptoms and Economic Relevance 

P. Heimberg, W. Adams, W. Hollberg, M. Holsteg, J. Winkelmann 

Chamber of Agriculture NRW, Cattle Health Service, Münster, 

Germany 

Background: In August 2006 the first outbreak of Bluetongue Disease 

(BT) in Middle Europe by Serotype 8 was confirmed: Most of the 

infected farms in Germany where located in Northrhine-Westfalia 

(NRW, about 800 farms). In 2007 we observed a rapid spread of BT 

virus in NRW, ending up with a total amount of nearly 10.000 positive 

tested farms, dairy cattle as well as sheep farms. The economic losses 

in sheep are quite easy to calculate because most of the infected 

animals died or have been culled (assumed mortality in NRW of about 

2%).The losses are compensated by the compulsory insurance for 

epizootic diseases and the government. In contrast dairy cattle did not 

show so many fatalities, but we have seen many symptoms with 

economic importance on the affected dairy farms. A questionnaire was 

sent to cattle farmers organized by the chamber of agriculture NRW to 

find out which BT symptoms are most important for the farmers. This 

was done to show if vaccination against BT is not only necessary for 

sheep but also for cattle to reduce casualities. 

Questionnaire: During the last week of November and first week of 

December 2007 we got answers from 565 farms. 527 of those were 

dairy farms. While 429 dairy farms where tested positive for BT 

Serotype 8, there were 98 which had no positive test result for BT. The 

farmers who answered to our questionnaire had in total 93.248 animals 

in their holdings. 

Results: We found out that two of three cattle farmers had direct losses. 

Answers resulted to an average morbidity of 10,5 %; the total amount 

of 725 losses lead to a letalitiy of 7,4 % and a mortality of 0,8 % by BT 

Serotype 8 infection in NRW. Abortion was detected up to first week of 

December 2007 concerned about 3 % of all dairy cows of BT-positive 

farms. The third important reason for economic losses by BT was seen 

in reduced amount of milk. Additionally 30 % of the farmers reported 

a significant increase in costs for veterinary treatments since summer 

2007. 

Conclusions: Although fatalities are not that much seen in cattle 

compared to sheep in NRW high remontation costs caused by fatalities 

and abortion, together with reduced milk delivery and increasing costs 

for veterinary treatment implemented that vaccination for cattle is as 

necessary as for sheep from point of economic view. At the end of the 

financial year 2007/08 in June 2008, casualities will be calculated for 

the average affected farm in Euro per cow. 

Key words: bluetongue, serotype 8, cattle, symptoms, economic 

relevance 

684 Comparative Field Clinical Efficacy of Florfenicol+Flunixine 

Meglumine in the Treatment of Bovine Respiratory Disease 

PR. Cloet 1 , O. Roy 2 , F. Pillet 2 , V. De Haas 3 , C. Maisonneuve 4 

1 Schering Plough Vétérinaire, Levallois Perret, France 

2 CEBIPHAR, CRO, Fondettes, France 

3 Schering Plough Animal Health, Levallois Perret, France 

4 Schering Plough Animal Health, Summit, United States 

The efficacy and safety of Resflor ® (florfenicol + flunixin meglumine) 

was evaluated in a GCP field trial as treatment of bovine respiratory 

disease in comparison with Draxxin ® (tulathromycin). 233 animals 

(260.5 98.21 kg) from 8 different farms were included with signs of 

bovine respiratory disease and allocated to 43 pens: 114 calves (21 pens) 

were given Resflor ® at a dose of 2 ml/15 kg and 119 calves (22 pens) 

were given Draxxin ® at a dose of 1 ml/40 kg. Treatments were 

administered on D0 by one subcutaneous injection. The nasopharynx 

was swabbed for bacteriological examination on D0 and upon failure. All 

animals included on D0 exhibited abnormal respiration, rectal 

temperature 40.3 °C, and depression. Rectal temperatures were 

measured, clinical signs scored, and injection sites observed on D0, 

D0+6h, and from D1 to D4. From D5 to D14, the animals were observed 

for clinical signs indicative of progressive disease (depression, abnormal 

respiration). The main pathogens isolated on D0 were Pasteurella 

multocida and Mannheimia haemolytica (6 clinical sites out of 8); 

Mycoplasma bovis and Histophilus somni were isolated in 2 farms. 

Rectal temperatures were lowest in the Resflor ® group from D0+6h to 

D4. The mean rectal temperature was statistically lowest on D1 in the 

Resflor ® group (P=0.0196, Mixed Linear model). Depression scores 

were statistically lowest in the Resflor ® group 6 hours after treatment, on 

D4, and on D8. Respiratory pattern was not different throughout the 

study between both groups. Success rate was highest in the Resflor ® 

group at each time point (from D5 to D14). Animals in both groups 

displayed injection site reactions throughout the study. One animal in the 

Resflor ® group died on D4 after having been removed on D3 

(worsening); gross pathology revealed lesions of severe acute 

bronchopneumonia and marked pulmonary emphysema related to BRSV 

infection. This clinical field study shows that both the test product and the 

reference product were efficient as treatment of severe bovine respiratory 

disease in cattle. Statistical analysis of the data shows a significantly 

higher efficacy of Resflor ® compared to Draxxin ® regarding the success 

rate of treatment and in abating high temperatures. 

685 Comparative Field Clinical Efficacy of a Single Intramuscular 

Injection of Marbofloxacine in the Treatment of Bovine 

Respiratory Disease 

O. Roy 1 , F. Pillet 1 , A. Heurtin-Vallé 2 , N. Frontczak 2 


2 Vétoquinol, Lure, France 

This multicentric field clinical trial aimed to assess the efficacy and the 

safety of Marbocyl ® S (marbofloxacine) as treatment of bovine 

respiratory disease in comparison with Draxxin ® (tulathromycin) and 

Nuflor ® (florfenicol). Two hundred cattle showing respiratory signs, 

depression, and rectal temperatures > 39.6 °C were included on 21 

farms. They were randomly allocated to three treatment groups and 

given one injection on D0 accordingly (8 mg marbofloxacine/kg, 2.5 

mg tulathromycine/kg, or 40 mg florfenicol/kg). Clinical signs were 

scored and rectal temperatures measured on D0, D1, D2, and D7. 

Pasteurella multocida, Mannheimia haemolytica, and Mycoplasma 

bovis were isolated on D0 from the animals in 14, 6 and 3 farms, 

respectively. Serological conversions related to natural infections with 

BVD and RSV viruses were detected in 1 and 5 farms, respectively. 

Rectal temperatures sharply decreased after treatment, mean values 

being lowest in the Marbocyl ® S group (P = 0,0096) on D1. Respiratory 

signs, general behaviour and appetite scores progressively decreased in 

all groups, with no significant difference between them. On D1, the 

success rate was significantly (P=0.0299 and P=0.0540) higher in the 

Marbocyl ® S (62.7%) group than in the other groups (< 50%). Success 

rates on D2 with Marbocyl ® S, Nuflor ® , and Draxxin ® were 

respectively at 83.1%, 82.6%, and 76.4% (no significant differences). 

Relapses were observed on D7 in 1.5%, 6.9%, and 9.2% of cases 

treated with Nuflor ® , Marbocyl ® S, and Draxxin ® , respectively, with 

no significant differences. Two animals died in the Draxxin ® group due 

to respiratory disease, and none in the Marbocyl® S and the Nuflor ® 

groups.The local tolerance was good in 96.5% of all the treated 

animals; 5, 1, and 1 animals displayed moderate signs of intolerance to 

Nuflor ® , Draxxin ® , and Marbocyl ® S, respectively. 

This clinical field study showed that the 3 tested products were efficient 

as treatment of respiratory disease in cattle. Marbocyl ® S induces cure 

significantly faster than the 2 other products tested products. 


686 Characterisation of Cattle Originated Escherichia coli 

O157:H7 Isolates by Randomly Amplified Polymorphic 

Dna(RAPD) and Antibiotic Resistance 

O. Aslantas 1 , Z. Cantekin 1 , B. Sareyyupoglu 2 , M. Akan 2 

1 

Mustafa Kemal University, Microbiology, Antakya, Turkey 

2 

Ankara Univercity Veterinary Faculty, Microbiology, Ankara, 

Turkey 

In this study, it was aimed to characterize 50 cattle originated 

Escherichia coli O157 isolates by randomly amplified polymorphic 

DNA (RAPD) analysis and their antibiotic resistance. RAPD analysis 

separated the E. coli O157 isolates into two clusters and 35 single 

isolates. All the strains were found to be resistant two or more of the 

antimicrobial agents tested. All the resistant strains were resistant to 

erytromycin and were suscaptible to amikacin, cefaperazone, 

imipenem and amoxycillin/clavulanic acid. 

Key words: E. coli O157, RAPD, Turkey 

687 In vitro Antimicrobial Susceptibility of Brucella abortus 

Isolated in Korea 

H. Eun Jeong, K. Jong Wan, H. Moon, C. Dong Hee, K. Sung Il, C. 

Yun Sang, H. In Yeong, J. Suk Chan 

National Veterinary Research & Quarantine Service, Animal Disease 

Research, Anyang-city, Korea, South 

Objectives of study: Brucellosis is the most common zoonotic 

infection worldwide. Recently, Brucella spp. have been recognized as 

a group of microorganisms that could be used as biological weapons of 

terrorism. Brucellae are intracellular bacteria, therefore treatments are 

very difficult. In Korea, isolated bovine Brucella strains are not tested 

for antimicrobial susceptibility. This study deals with the drug 

susceptibility of naturally occurring Brucella abortus strains. 

Materials and Methods: During 1998 to 2006, B. abortus strains were 

isolated from dairy and beef cows (Hanwoo). The districts from which 

the strains were isolated are eight areas. The colonies of eighty four 

strains were tested for biological characteristics. Then, these strains 

were evaluated by means of inhibition study, minimal inhibitory 

concentrations (MICs) performed by method of broth microdilution 

susceptibility test. 

Results: Tetracyclines demonstrated the highest activity of any 

antimicrobial agent in this study (MIC range; 0.063~0.5 mg/mL). Among 

tetracyclines, minocycline had greater activity with MIC 50 and MIC 90 of 

0.125 mg/mL. The MIC 50 of tetracycline and doxycycline was same as 

0.25mg/mL and the MIC 90 of these two drugs are also same as the MIC 50. 

Four quinolones, ofloxacin, norfloxacin, enrofloxacin and ciprofloxacin 

showed high activity against B. abortus. The MIC 50 of ciprofloxacin was 

0.25 mg/mL and MIC 90 was 1 mg/mL. The MIC 50 and MIC 90 of 

ofloxacin and enrofloxacin were 0.5 mg/mL and 1 mg/mL, respectively. 

However, the MIC 50 and MIC 90 of norfloxacin was 4 mg/mL, which was 

less activity against B. abortus than other quinolones. Streptomycin and 

erythromycin had effective activities with MIC range 0.5~2 mg/mL and 

1~4 mg/mL respectively. Rifampin had a good effect with MIC range 

from 0.5 to 4 µg/mL. The combination of tetracycline and rifampin 

showed good effect with MIC range from 0.063 to 0.5 mg/mL. 

Conclusions: The choice of antibiotics in this study focused on in vivo 

application to humans. We observed variation in sensitivity among B. 

abortus isolates in Korea. Tetracyclines are the most effective drugs of 

all other drugs measured. And aminoglycosides and quinolones 

showed active effect against B. abortus strains isolated in Korea. The 

combination of tetracycline and rifampin was used in some clinical 

reasearch. This study also demonstrated active effect of combination of 

tetracycline and rifampin. 

Key words: Brucella abortus, antimicrobial susceptibility, minimal 

inhibitory concentrations (MICs) 

688 Persistent Infection of Foot and Mouth Disease Virus in some 

Slaughtered Cattle and Buffalos in Ahwaz Abattoir by RT- 

PCR 

M. Ghane 1 , A. Rasoli 2 , M. Seyfiabad Shapouri 3 , M. Haji Hajikolaei 2 , 

A. Komeilian 4 

1 School of Veterinary Medicine, Shiraz Univrsity, Shiraz, Iran, 

Department of Clinical Science, Shiraz, Iran 

2 Faculty of Veterinary Medicine, Shahid Chamran University, 

Department of Clinical Science, Ahwaz, Iran 


3 Faculty of Veterinary Medicine, Shahid Chamran University, 

Department of Pathobiology, Ahwaz, Iran 

4 Islamic Azad University, Science and Research Branch, Department 

of Clinical Science, Tehran, Iran 

Foot and Mouth Disease (FMD) is a highly contagious disease of 

cloven-hoofed animals and has great potential for causing severe 

economic losses. An important feature of FMD in ruminants is 

persistent infection that also called carrier state.This may occur in FMD 

convalescent period as well as in vaccinated ruminants following 

exposure to infectious virus. In order to evaluate prevalence of 

persistent infection of FMDV in cattle and buffalos slaughtered in 

Ahwaz abattoir, 200 head of mature female cattle and buffalos were 

studied. After slaughter, samples were collected from oropharyngeal 

fluid. RT- PCR was carried out with general primers for all major virus 

serotypes. Out of 100 cattle samples, none of them were positive for 

virus but out of 100 buffalos samples, two positive samples were 

observed. Since buffalos are in close contact with other sensitive 

animals such as cattle and sheep, they may be of great importance in 

spreading the disease in the area. 

689 Prevalence of Bovine Herpesvirus Type 1 in Southern Backa 

and Srem District in the Republic of Serbia 

S. Lazic 1 , T. Petrovic 1 , D. Lupulovic 1 , M. Maljkovic 2 , D. Bugarski 2 

1 Scientific Veterinary Institute Novi Sad, Virology, Novi Sad, Serbia 

2 Scientific Veterinary Institute Novi Sad, Department of 

Epizootiology, Novi Sad, Serbia 

Determination of the prevalence of bovine herpesvirus type 1 (BHV-1) in 

Southern Backa and Srem district was carried out by determining specific 

antibodies against BHV-1 using commercial ELISA test (Test Line Ltd, 

Check Republic). The total number of cattle in these two districts was 

123911, but the examination comprised 22139 animals (17.87%). The 

estimation of prevalence was done using software program Win Epscope 

version 2. The prevalence of infection was calculated for every 

municipality, and then for the district. In Southern Backa district from the 

total number of 52383 the examination comprised 11911 animals from all 

12 municipalities. The BHV-1 seropositive animals were detected in all 

the municipalities, except in Sremski Karlovci. In this municipality out of 

114 animals, 35 were examined and all were seronegative. In other 

municipalities the percent of seropositive animals varied and ranged from 

11.84% (Temerin municipality) to 33.46% (Becej municipality). In most 

municipalities (5) the percent of BHV-1 seropositive animals ranged from 

11.84% to 20%. In 3 municipalities the percent of seropositive animals 

was 25%, and it was above 30% in 3 municipalities. In Srem district the 

examination was carried out in all 7 municipalities, so out of 71,548 

animals 10,228 were examined. The smallest number of seropositive 

animals was discovered in Ruma municipality (14.57%), and the largest 

in Pecinci municipality (23.33%). In two municipalities (Sid and Sremska 

Mitrovica) the percent of seropositive animals was lower than 20%, while 

in other municipalities (Indija, Irig and Stara Pazova) the percent of BHV- 

1 seropositive animals ranged from 21.60% to 22.20%. Based on these 

results it may be concluded that in Southern Backa district out of 52363 

cattle there were 11911 (22.75%) examined on BHV-1. The specific 

antibodies against BHV-1 were detected in 3123, what means that the 

prevalence was 26.22%. In Srem district out of 71548 animals there were 

10228 examined animals (14.29%), but specific antibodies against BHV- 

1 were detected in 2045 animals what means that the prevalence was 

19.99%. 

Key words: BHV-1 prevalence, ELISA, Serbia 

690 Importance of BHV-1 in Etiology of Bovine Respiratory 

Disease Complex in Calves in Province Vojvodina (Republic 

of Serbia) 

D. Bugarski 1 , T. Petrovic 2 , I. Pusic 1 , D. Lupulovic 2 , D. Milanov 3 , 

S. Lazic 2 

1 

Scientific Veterinary Institute Novi Sad, Department of 

Epizootiology and Health Protection of Ruminants, Novi Sad, Serbia 

2 

Scientific Veterinary Institute Novi Sad, Department of Virology, 

Novi Sad, Serbia 

3 

Scientific Veterinary Institute Novi Sad, Department of 

Microbiology, Novi Sad, Serbia 

In Serbia eradication of BHV-1 infection has been voluntarily carried 

out, so prevalence ranged from 5% to more than 50% depending on the

area. The importance of BHV-1 in etiology of respiratory disease 

complex in beef calves was conducted in the years 2005-2007. It was 

carried out on 12 fattening farms (250-500 animals per herd) by 

determining seroconversion after enzootic pneumonia outbreak, 

examining the presence of antibody titer before slaughtering and the 

virus isolation from nasal swabs was also done at the beginning of an 

acute respiratory infection. A characteristic feature common for all the 

farms is that new animals are introduced several times a year and 

therefore it is impossible to avoid simultaneous presence of different 

aged animals, what gives a possibility for long retention of the virus in 

a herd. Beef farms in Vojvodina purchase young calves from mountain 

regions, where the prevalence of BHV-1 in adult animals is even 45%. 

On all the farms the animals were clinically examined; out of total 

number the blood was sampled from 3-10%, most of them were sick, 

chronically sick or recovered from the respiratory syndrome. The 

second sampling, for determining seroconversion, was carried out 3-4 

weeks after the first one. In all the fattening farms enzooty of 

bronchopneumonia was present. Clinical symptoms were: increased 

body temperature, weak serous nasal discharge, coughing and 

dyspnoea. BHV-1 seroconversion in totally recovered and animals 

chronically ill from bronchopneumonia was not determined in all the 

examined herds. In newly arrived calves seroprevalence on BHV-1 

ranged from 0-10.56%. At the end of fattening period antibodies 

against BHV-1 ranged from 0-100%, depending on a herd. However, 

BHV-1 was not isolated during acute infection. From the obtained 

results it may be concluded that, currently, BHV-1 is not of great 

importance for developing respiratory syndrome in the herds of beef 

calves, what was not the case in past decades, so the infection is 

subclinically present. In fact, in all the beef farms, after the recovery 

from the disease, immuno-response towards BRSV and Pi-3 was 

determined. In the lungs of dead animals only mycoplasma was 

determined, or mycoplasma with bacteria. 

Key words: BHV-1, beef, respiratory syndrome, Vojvodina 

691 Effect of Vaccination against Leptospira on Dairy Farm 

Reproduction Indexes 

D. Ruiz Di Genova 

COVAP, Bovine Technical Services, Pozoblanco, Spain 

Leptospirosis is a worldwide occurring disease, that when caused by 

serovars adapted to dairy cows (serovar Hardjo) produce chronic 

infections, where the signs are less evident. 

Objective: To assess the effect of vaccination against Leptospira 

interrogans serovar Hardjo on the reproduction indexes. 

Materials and Methods: The study includes 9 dairy farms (mean 70 

cows/farm), in southern Spain with complete reproductive records 

comprising a year before the first leptospira vaccination and a year 

afterwards. In seven farms, serological diagnosis of leptospirosis was 

performed by MAT (microagglutination test) for serovar hardjo, prior 

to vaccination. A positive farm was considered when presenting one 

animal with a titter > 1/10. Another two farms were vaccinated due to 

the clinical suspicion, after ruling out other reproductive diseases.The 

monthly conception rates and first service conception rates were used 

as fertility indexes, comparing results obtained one year before 

vaccination with results obtained on the year afterwards.Cumulative 

data were analyzed using logistic regression, which allows to establish 

a model for dicotomic results (pregnant/not pregnant). The result is 

expressed as an odds ratio indicating the different occurrence 

probability of one or the other event, that is, the probability of 

pregnancy after one service, before and after vaccination. 

Results: The probability of pregnancy in vaccinated cows was 1,25 

times higher than in not vaccinated ones. (OR=1,25399, EE=0,0673; 

P=0,0008), with the “vaccination” variable accounting for 21% of this 

variation in fertility. In the first-calving cows the probability of 

conception increased 1,70 times after vaccination (P

using EPFC. The intradermal tuberculin comparative test was 

applied to all animals at 12 and 24 months postvaccination. IFN 

production of cell cultures stimulated with bovine PPD showed 

significant increase (1.4, 0.90, 1.4 OD) at days 15, 90 and 180 

postvaccination, in the group innoculated with EFPC. In the group 

vaccinated with BCG the increase was observed at 60 and 150 days 

postvaccination (0.90, 1.7 OD) while the control group showed 

increment at day 90 (1.35 OD). Animals from the three groups 

showed an increse (1.4 OD) at day 60 when stimulated with avian 

PPD. Stimulation with ESAT 6 did not give a clear response in any 

of the groups. With respect to the humoral response, all groups 

showed an increase in OD at day 90 but was significantly higher 

(P

esponse in the eye produces accumulation of polymorphonuclear 

neutrophils (PMNs) in tissue and ocular fluids, increase of the vascular 

permeability and therefore an increase of AH proteins. The increase of 

PMNs explains the larger concentrations of TMS reached in AH of the 

animals with keratoconjunctivitis. Although TMS levels in AH (either at 

5 or 10 mg/kg) persisted during 72-96 h, they never surpassed the M. bovis 

MIC (< 1 µg/mL). Nevertheless a longer TMS persistence was evident in 

sick animals, being documented through longer mean residence time 

(41.2, 45.7 and 22.7 vs 15.9 h for the G1, 2, 3 and 4 respectively) and a 

smaller clearance from AH (222.8, 469.5, 773.3 vs 1613.5 mL/h/kg for 

the G 1, 2, 3 and 4, respectively). These information suggest that TMS 

penetrate to the ocular balloon and concentrates in AH depending on the 

degree of inflammation. Concentrations in vitreous humors and ocular 

tissue, should be evaluated in the future to consider the use of the TMS for 

the treatment of certain ocular infections. 

Key words: tilmicosin, aqueous humor, keratoconjunctivitis, 

pharmacokinetic, cattle 

697 The use of PCR for Identification of Brucella abortus in 

Samples from Aborted Fetuses in a Dairy Farm in the State of 

Aguascalientes, Mexico 

D. Limón 1 , G. Martínez 1 , G. Zenteno 1 , B. Arellano 2 , E. Diaz 1 , 

R. Flores 1 , F. Morales 1 

1 INIFAP, CENID Microbiologia, MEXICO, D.F., Mexico 

2 Universidad Nacional Autonoma de México, Facultad De Medicina 

Veterinaria, MEXICO, D.F., Mexico 

Abomasal fluid and tissues conserved in 10 % bufferated formalin 

solution from 13 aborted fetuses, as well as sera from 55 aborted cattle 

were collected in a dairy farm located in Aguascalientes, Mexico and 

were transported to the Diagnostic Laboratory of CENID- 

Microbiologia, INIFAP. The abomasal fluid samples were inoculated 

in enriched Farrel’s medium in order to attempt the Brucella abortus 

isolation. The tissues samples were processed for histophatological 

analysis, while the sera were used for detection of BVD, IBR, 

Neospora, Leptospira and Brucella antibodies. Those Brucella spp 

suggested isolations were examined by using a PCR procedure. 

Depending of the amplified fragments (361, 456, 1063 and 1298 bp) it 

is posible the identification and differentiation of field and vaccinal 

Brucella abortus strains. Six isolated strains were cultured from the 

abomasal fluid and all of them were identificated as Brucella abortus 

field strains by PCR. Brucellosis typical lesions (multifocal necrotical 

hepatitis and exudative pneumonia) were observed in 4/13 studied 

fetuses. The result of serological examination of 55 samples was as 

follows: Brucellosis, 27 %; Neosporosis, 40 %; Leptospirosis, 65 % 

(titers > 1:400) and IBR, 100% and BVD 80 %. During the last few 

years Leptospirosis and Neosporosis have shown an increase in their 

importance as a cause of abortion in cattle in Mexico, our results in the 

present study confirm this affirmation. The fact that a specific 

vaccination program to prevent neosporosis does not exist in the dairy 

farm under investigation, makes it important to consider this agent as 

important causal of the abortion. In regards to Brucellosis, it is a 

disease with high prevalence in the dairy cattle farms despite the fact 

that there is an intensive official control campaign in Mexico and 

excellent vaccines such as S19 and SRB51 exist; for this reason it is 

important to have diagnosis techniques with high sensivity and 

specificity as it is the case of PCR technique. It is important to promote 

the use of integral etiological diagnosis of abortions in order to be in 

conditions to apply adequate and effective control measurements. It 

often occurs that in some laboratories in Mexico the samples are tested 

only for diagnosis of brucellosis and do not consider the use of 

techniques for determinate the possible presence of other 

microorganisms associated with reproductive problems. 

Financial support SAGARPA-CONACyT. (2004/COI/23) 

Key words: PCR, Brucella abortus, aborted fetuses 

698 Long-Term Excretion of Brucella abortus, Strain B19 in Milk 

and Urine by Vaccinated Adult Female Bovines 

M. Genovez 1 , W. Pacheco 1 , C. Pozzi 2 , L. Paulin 1 , M. Gamberini 3 , 

S. Azevedo 4 

1 Instituto Biológico, Animal Health Center, Sâo Paulo, Brazil 

2 Instituto de Zootecnia, Animal Health Center, Nova Odessa, Brazil 

3 Universidade Federal de Goiás, Animal production, Goiania, Brazil 

4 Universidade Federal De Campina Grande, Department for 

Infectious diseases- Epidemiology, Patos, Brazil 

Abortion caused by Brucella abortus is a common bovine 

reproductive disease in many countries and may be currently 

prevented by vaccination with live B19 vaccine, administered to 3-8 

month old heifers. The length of excretion, its relation to the 

hormonal cycle and its effect on contactant and susceptible animals, 

including man, are not completely known. The persistence of B19 

excretion in milk and urine by adult dairy cows vaccinated at 3-8 

months old was investigated in this study. Samples were collected 

during 12 months (15 collections), beginning at estrus until 

immediate post-partum, from 14 females in three groups: G1(3-4 

years old), G2 (5-6 years old) and G3 (7-9 years old). Milk and urine 

samples were monitored by bacteriologic analyses and Multiplex 

PCR differential for B.abortus and B19 strain; blood serum samples 

were analyzed by Rose Bengal test for brucellosis diagnosis and 

direct estradiol ELISA KIT and progesterone ELISA KIT 

(Diagnostics Biochem., Canada) were used for hormone monitoring. 

All milk samples were negative in serology and bacteriologic 

examination. B19 was detected by PCR in 74/210 (35.2%) and 

12/210 (5.7%) from urine and milk samples, respectively (p=0.001). 

Excretion mainly occurred from the beginning of estrus to 150 days 

of pregnancy and for 30 days after parturition. Hence, excretion was 

intermittent, no matter the age of the animal and the time after 

vaccination. Hence, excretion was intermittent, no matter the age of 

the animal, the time after vaccination, and occurred in cows up to 

nine-year old. Hormonal stage did not seem to affect B19 excretion, 

and may be associated with erythritol scarcity. The impact of longterm 

B19 excretion requires more attention. B19 may remain in the 

environment with a real possibility of transmission to negative cattle 

herds, mainly to bulls, or even to other susceptible animal species, 

including man. Urine was the best sample for B19 detection. 

This paper was supported by FAPESP: Fundaçao de Amparo a 

Pesquisa do Estado de Sao Paulo, Brazil 

Key words: bovine brucellosis, B19 excretion, hormonal cycle 

699 Main Causes of Abortion in Cattle at Different Areas of 

Mexico 

G. Zenteno 1 , F. Manrriquez 1 , I. Vitela 2 , L. Diaz 4 , R. Flores 1 , 

D. Martinez 3 , F. Morales 1 

1 

INIFAP, CENID Microbiologia, México DF, Mexico 

2 

Instituto Tecnológico el Llano, Microbiología, Aguascalientes, 

Mexico 

3 

Universidad Veracruzana, Facultad de Medicina Veterinaria y 

Zootecnia, Facultad De Medicina Veterinaria, Veracruz, Mexico 

4 

Gobierno del Estado de Aguascalentes, Comité de Fomento y 

Protección Pecuaria, Aguascalientes, Mexico 

Data related to the incidence and prevalence of etiological agents 

associated with reproductive failures in cattle of Mexico are the result 

of isolated studies, only a few cases include results of integrated 

diagnosis. Some papers describe the results of serological profiles, but 

in most of them the interpretation of the results does not permit the 

diagnosis of the etiological agent. For example, previous studies 

performed by our research group demonstrated different prevalences 

of brucellosis, BVD, IBR, neosporosis and leptospirosis as follows: 

The brucellosis prevalence was always high (12-21 %) except in 

Veracruz (3%); Leptospirosis was similar to brucella with the highest 

prevalence in Aguascalientes (27%) and the lowest in Veracruz (3%). 

The BVD and IBR highest prevalences were observed in the same 

states (56-86%) while in Veracruz there were lower (47 and 38 % 

respectively). The prevalence of neosporosis varied between 30 and 45 

% in most states, but in Veracruz was only 15 %. With these results it 

was impossible to relate the abortions with a particular infectious agent. 

The purpose of this study was to perform an integral, comparative 

serological profile, in sera collected from aborted and non aborted 

animals from 7 dairy cattle in different areas of the state of 

Aguascalientes, Mexico. Blood samples were collected from cattle 

aborted during a five month period previous to the collection and from 

non aborted animals. All samples were tested for the identification of 

Brucelosis, BVD, IBR, Neosporosis and Leptospirosis antibody titers. 

The results of this serological study, were then used in order to compare 

the prevalences of each etiological agent with either, aborted and non 

aborted populations. The results of this comparative study 

demonstrated that in some farms more than one etiological agent was 

found to be important: in 4 cattle farms brucellosis was a serious 

problem; in one farm the main cause of abortion was IBR, while BVD 

and neospora appeared to be the most important causes of abortion in 


three cattle farms each. Leptospirosis was also present. It is notorious 

the fact that in dairy cattle different etiological agents could be 

involved in reproductive problems, therefore the diagnosis procedures 

must be integral, including a set of serologic test that allows to 

determinate their presence and to be able to apply adequate and 

effective control measures. 

Financial support: SAGARPA-CONACyT. (2004/COI/23) 

Key Words: abortion, cattle, serology 

700 Viability of Mycobacterium Avium Subsp. Paratuberculosis 

(MAP) in Baled Grass Silage 

J. Khol 1 , V. Beran 2 , P. Kralik 2 , M. Trckova 2 , I. Pavlik 2 , 

W. Baumgartner 1 

1 Clinic for Ruminants, Department for Farm Animals and Herd 

Health, Vienna, Austria 

1 Veterinary Research Institute, Brno, Czech Republic 

Objective of the study: Ensiling of grass from areas where livestock or 

wild animals infected with paratuberculosis are grazing can be a 

possible source of Mycobacterium avium subsp. paratuberculosis 

(MAP) infection in domestic ruminants. The results of a preliminary 

study about the viability of MAP, the causative agens of 

paratuberculosis, in baled grass silage should be presented in this 

poster. 

Materials and Methods: Seventeen bales of grass silage were spiked 

with a suspension containing MAP. Silage samples were collected 

periodically for four months to observe MAP viability over time. 

Collected samples were tested for MAP by bacterial culture and Real 

Time-PCR (Polymerase Chain Reaction). 

Results: Viable MAP could not be detected at any sampling date 

during the trial by culture, more than 60 % of the analysed samples 

were tested positive by Real Time-PCR. 

Conclusions: Even though the results of the presented work suggest 

that grass silage seems to be an unlikely source of paratuberculosis 

infection for livestock, further investigations are needed to elucidate 

the reaction of MAP to unsuitable environmental conditions and their 

influence on the infectivity of the bacterium. 

701 Phylogenetic Position of an Unreported BPV Type Detected 

From a Cutaneous Lesion of a Brazilian Cattle Herd 

M. Lunardi, M. Claus, A. Alfieri, A. Alfieri 

Universidade Estadual de Londrina, Departamento de Medicina 

Veterinária Preventiva, Londrina, Brazil 

In Brazil, Bovine Papillomavirus (BPV) infections are endemic in 

beef and mainly in dairy cattle herds. Despite the high frequency of 

BPV infection, the identification of BPV types in Brazilian cattle is 

still sporadic. In a prior study, through the analysis of a partial 

segment of L1 gene, we could identify the presence of previously 

described BPV types and four putative new BPV types associated 

with skin warts. The aim of this study was to determine the entire L1 

nt sequence of a putative novel BPV type (BPV/BR-UEL2) detected 

in Brazil, and thus state its phylogenetic position. As a phylogenetic 

analysis employing the FAP amplicon had revealed our isolate as 

closest related to BPV-4 (Xi genus), two pair of degenerate primers 

were designed by using alignments of L1, L2, LCR regions of 

genome of Xi genus representatives. In addition, aiming to obtain the 

full L1 gene sequence, the previously described FAP primer pair was 

also employed both in the original form as in combination with 

designed primers. The PCR amplicons were purified from agarose 

gel and submitted to cloning. Plasmid DNA from 2 clones was 

sequenced in both directions using M13 forward/ reverse primers. 

Sequences were examined with the software PHRED for quality 

analysis and the consensus sequence was determined using the 

software CAP3. The alignment was obtained with the software 

BioEdit. A neighbour-joining phylogenetic tree was constructed 

with complete L1 ORF sequences of 38 PVs classified in 18 genera, 

and with the entire L1 ORF sequence of BPV/BR-UEL2 isolate, 

using the MEGA program v.3.1. A consensus sequence, representing 

the terminal portion of L2 gene, the complete sequence of L1 gene 

and an initial segment of LCR, could be achieved from three PCR 

amplicons. By ORF analysis, it was possible to determine that the L1 

ORF encoded protein of the Brazilian isolate consists of 532 aa. The 

phylogenetic analysis revealed that the BPV/BR-UEL2 is related 

with BPV types held in Xi genus. Besides, this isolate displayed the 


highest L1 nt sequence similarity with BPV type 4 (74%), suggesting 

its classification in the Xi genus. The realization of further studies 

involving the molecular epidemiology of BPV infections, in 

Brazilian cattle herds as much in diverse geographical areas around 

the world, become necessary to verify the prevalence of this new 

viral type and to check its association with cutaneous lesions. 

Financial support: CNPq, CAPES, FINEP and FAP/PR 

Key words: BPV; L1 ORF; phylogeny 

702 Monitor pProject : Evidence of Some Respiratory Virus 

Isolation from Marchigiana Breeding Farms with Respiratory 

Disorders, Note 2. 

S. Petrin 1 , M. Panicci 1 , S. Briscolini 1 , L. Cucco 1 , M. Ferrari 2 , 

G. Filippini 1 , G. Pezzotti 1 

1 

Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, 

Perugia, Italy 

2 

Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia 

Romagna Brescia, Brescia, Italy 

Objectives of study: In the framework of the monitor project, 

necropsy has been carried out on 52 dead calves originating from 

herds with bovine respiratory disease (Monitor project note 1). This 

report describes the results of the bacteriological and virological 

investigations carried out on subjects died for respiratory disorders. 

Materials and methods: Organs (trachea, lung, bronchial lymph nodes), 

were collected from 52 dead animals for bacteriological examination and 

viral isolation. The bacteriological examination was performed according 

to Quinn (1999); briefly, specimens were streaked on blood agar (BA), 

Mc Conkey agar (MC), Mannitol Salt agar (MSA); and incubated for 12- 

24 h at 37 °C. Isolates were characterized according to Quinn (1999), and 

further identified using the API Systems (Biomerieux). Testing for 

Mycoplasma spp. was carried out inoculating PPLO agar and broth, 

incubated for 12 days at 37 °C under 5% CO 2, and checked daily. For 

virus isolation the samples were diluted, centrifuged and the supernatants 

were inoculated into BEK cells. In absence of citopathic effect (CPE), 3 

subpassages were made. In positive cases, the identification of the isolated 

virus was evidenced with serum neutralization test (SN) using reference 

immune serums against IBR and with nested-PCR methods according to 

Vilcek S. et al. (1994) to identify BRSV. For the SN tests virus isolates 

were diluted serially and mixed with the reference immune serum in 96well 

microtiter plates which were held for 90 min at 22 °C. After the 

incubation 20,000 BEK cells in E-MEM were added. The plates were 

checked for 7 days, and the CPE was evaluated. Virus dilution mixed with 

E-MEM represented the control. The virus was identified if titre in the 

presence of immune serum was at least 2 log lower than the virus titre in 

E-MEM. 

Results: The results evidenced the presence of the following bacteria: 

M. haemolytica (15 %); P. Multocida (26 %); Mycoplasma spp (4 %); 

Negative cases (55 %). From trachea and lung samples of three 

animals, CPE was detected in BEK cell cultures whose features were 

typical of IBR or BRSV, respectively. They were detected at the first or 

second serial passage. IBR and BRSV were identified by SN and nested 

PCR tests, respectively. 

Conclusions: The results of the investigation demostrated that viruses, 

isolated from Marchigiana breeding farms with respiratory disorders 

are members of the Herpesviridae and Paramyxoviridae family. 

703 Previously Described BPV Types and Putative New Types in 

Cutaneous Papillomatosis from Brazilian Cattle Herds 

M. Claus, M. Lunardi, A. Alfieri, A. Alfieri 



The aim of the current study was to report the identification of 

putative new BPV types in skin warts in cattle herds from Paraná 

state of Brazil. Papilloma specimens (n=27) were taken individually 

from diverse body sites of adult and young bovines, from dairy (n=2) 

and beef (n=2) cattle herds from Paraná state, South region of Brazil. 

The PCR assay was carried out using the primer pair FAP59 and 

FAP64. All PCR products were purified and a direct sequencing was 

performed with FAP primers. For the amplicons which the prior 

analysis revealed them as a putative new BPV type, a cloning and a 

further sequencing, in both directions, was performed employing the 

plasmid DNA from two selected clones of each sample. For quality 

analysis of chromatogram readings and determination of the

consensus sequences, the PHRED and CAP3 softwares were used, 

respectively. The sequence identity was verified with all sequences 

deposited in the GenBank by using the BLAST software. The 

alignment was obtained with the software BioEdit, and a neighbourjoining 

phylogenetic tree was constructed using the MEGA program 

v.3.1. A new PV type is defined as a complete genome, whose L1 

gene sequence is at least 10% dissimilar from the closest known PV 

type. As the FAP59/64 products represent only part of the L1 gene, 

the obtained sequences that had differed from BPV-1 to -10, were 

named putative new PV types. PV DNA was detected in all DNA 

samples analysed, resulting in amplicons with 478 bp. The BPV-1, - 

2, -6 and –8, were identified in 4, 8, 9, and 2 papilloma specimens, 

respectively. In addition, four putative new BPV types were 

identified in other six skin warts. Four Brazilian putative new BPV 

types were designated as BPV/BR-UEL2 to -5 (accession numbers: 

EU293538, EU293539, EU293540 and EU293541, respectively). As 

our putative new BPV types were detected from skin warts, their 

potential for causing cutaneous lesions could be suggested. Besides, 

the BPV/BR-UEL4 and -5 isolates represent the first putative BPV 

types closely related with BPV-2 and BPV-8, respectively. The 

detection of these BPVs types in skin wart specimens supports the 

existence of virus diversity throughout the Brazilian livestock. 

Finally, the identification of four putative new BPV types is the first 

report of the presence of different BPV types in American continent. 


Key words: BPV, skin wart, phylogeny 

704 Monitor Project : Serological Survey in Marchigiana 

Breeding Farms, Note 1. 

M. Paniccia’, G. Filippini, A. Duranti, F. Ciuti, A. Dorinzi, P. Papa, 

P. Mangili, S. Petrini 

Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, 

Perugia, Italy 

Objectives of study: In order to evaluate the prevalence of the major 

infections causing respiratory, enteric or reproductive syndromes in the 

Marchigiana breeding farms of the Marche region (Italy), a 

surveillance programme, named Monitor, was carried out through 

serological investigations. The following agents were considered: 

BHV-1, BRSV, PI-3 and BVDV. Moreover, the presence of antibodies 

against Mycobacterium avium subspecies paratuberculosis (MAP) was 

evaluated. Within the reproductive infections, Chlamydia abortus 

(CA), Leptospira pomona (LP), Leptospira hardjo (LH) and Neospora 

caninum (NC) were considered. 

Materials and methods: A cross sectional study was carried from March 

to December 2006. The target population consisted of bovine farms for 

meat production present in Marche region (49.737 heads and 2.802 herds). 

A one-stage cluster sampling was performed and 22 herds were selected. In 

these farms, 1662 bovine serum samples from adult cattles (>12 months of 

age) were collected and tested against BHV-1, PI-3, BRSV, BVDV, MAP, 

NC using commercial ELISA tests, CA through complement fixation test 

(Donn A et al., 1997) and LP-LH through microagglutination test (O.I.E. 

Methods). The results were evaluated using Epi-info 3.4.2 to estimate the 

prevalence of the infections considered. 

Results: The result demonstrated that all the agents considered were 

present in the Marche region. In particular, the estimate prevalences were 

34,0 % (I.C.95%: 6,7 -61,3) for BHV-1, 91,9 % (I.C.95%: 83,6 -100 %) for 

PI3 and 54,9% (I.C.95% 37,0 - 72,8%) for BRSV. Moreover, 3,5 % (I.C. 

95%: 0,1 - 6,4%) of the animals resulted positive for antibodies against 

MAP, 3,8 % (I.C.95%: 2,8-6,1%) for CA, 7,4% (I.C. 95%: 2,3 - 12,6%) for 

NC and 1,5% (I.C. 95%: 1,3-4,3%) for LP-LH. Finally, 1,1% (I.C. 95%: 

0,4-2,6%) resulted positive for the presence of BVDV in blood. 

Conclusions: The results demonstrated that the agents considered 

largely occur in cattle population of the Marchigiana breed and must be 

taken in consideration approaching the diagnosis of respiratory, enteric 

or reproductive syndromes in these kind of farms. In addition, the 

results have to be assumed as a baseline for further investigations or for 

the definition of future intervention measures in this traditional Italian 

bovine breed, such as vaccination or control or eradication programmes 

for selected infectious agents. 

705 Botulism in Cattle: how the Laboratory can Help in 

Diagnosing the Disease? 

F. Gessler 1 , H. Boehnel 2 

1 Miprolab GmbH, Goettingen, Germany 

2 

Institute for Applied Biotechnology in the Tropics, Goettingen, 

Germany 

For almost centuries botulism is a disease complex known to the cattle 

farmer. It is caused by Clostridium botulinum, an anaerobic, sporeforming 

bacterium. It is not the bacterium itself, but the highly potent 

neurotoxins, which at the end are responsible for the clinical signs 

observed. According to their chemical structures and pathophysiological 

properties seven distinct toxin groups were identified, 

which were named A to G. Types C and D are considered to be those 

toxins predominant in cattle botulism leading to flaccid muscular 

paralysis. The classical form of botulism is feed-poisoning, an 

intoxication caused by the uptake of the neurotoxin pre-formed in the 

feed, eg silage. In Botulinum toxico-infections, spores and/or bacteria 

are ingested, colonize the intestine and produce the toxins in vivo. It is 

the toxico-infectious botulism, which has been re-emerging over the 

recent years. Besides clinical, pathological and epidemiological 

investigations, the detection of the toxins and/or the toxigenic bacteria 

in the laboratory contributes to a valid diagnosis of the disease. 

The presentation will cover: 1. basic aspects of botulinum neurotoxins, 

2. selection of samples and matrices for the laboratory investigations 

(eg: feed, soil, clinical samples), 3. short overview on the detection of 

the toxins, specific antibodies and the bacteria in the small clinical or 

the specialized microbiological laboratory, and 4. interpretation of the 

results and recommendations 

706 Application of the MTT Assay for Titration of Cytopathic 

Bovine Viruses 

A. Simsek, O. Bulut 

Faculty of Veterinary Medicine, The Selçuk University, Department of 

Virology, Konya, Turkey 

In the current study, a colorimetric method for titration of cytopathic 

bovine viruses was developed. MTT colorimetric assay is based on the 

ability of a mitochondrial dehydrogenase enzyme from viable cells in 

culture to reduce the tetrazolium salts of the yellow MTT and form a 

blue formazan crystal. Bovine Herpesvirus Type 1 (BHV-1), 

Parainfluenza 3 (PI-3), Bovine Rotavirus (BRV) and Bovine Adenovirus 

Type 1 (BAV-1) which are cytopathic bovine viruses were propagated 

and titrated in Madin Darby bovine kidney (MDBK) cell line in this 

study. All viruses were also assayed for titration with the MTT 

colorimetric method. The correlation between virus titers when two 

assays were determined by assessing the correlation coefficient (r) with 

Microsoft Excel program. The MTT colorimetric assay was showed a 

good correlation in terms of the viable cell number and the reduction of 

MTT. The titers determined by the two methods (microscopic 

insemination for CPE and MTT microassay) were found to be highly 

correlated (r=0.995). In conclusion, the results of this study confirmed 

that the MTT assay can be applied to titrating viral systems in which the 

virus is able to produce CPE in vitro. In particular, the MTT method 

proved to be useful to estimate cell densities in virus-infected cell 

cultures and was more effortless than classical methods. Concurrently, 

the method could be computerized with an ELISA reader and facilitated 

the titration of large numbers of virus samples in a short time. 

Key words: viral titration, colorimetric assay, cattle 

707 Comparative Study of Rose Bengal and Competitive ELISA 

Tests in Double Purpose Herds in the State of Lara - 

Venezuela 

J. Maldonado 1 , A. Kowalski 1 , C. Villasmil 1 , O. Verde 2 , M. Milla 3 , 

J. Alvarado 1 , F. Escalona 1 , A. Rodriguez 1 

1 

Universidad Centroccidental Lisandro Alvarado, Barquisimeto, 

Venezuela 

2 

CIEPE, San Felipe, Venezuela 

3 

UNEY, San Felipe, Venezuela 

Brucellosis is a zoonotic disease that negatively affects milk production 

in Latin America. Infections with Brucella abortus bacteria observed in 

human populations are fundamentally caused by ingestion of nonpasteurized 

milk by-products. In Venezuela, the most of the milk 

production is obtained from crossbred double purpose herds, which are 

generally under deficient sanitary conditions. A significant amount of 

milk produced is used for production of non-pasteurized cheese for 

human consumption. In Venezuela, the official screening test for 

brucellosis is the Rose Bengal test (RBT), leaving competitive ELISA 


(cELISA) only to definitively confirm results obtained in the 

aforementioned test. It performed a trial study to assess the status of 

brucellosis in the state of Lara, Venezuela. Initially, milk ring tests 

were performed in 320 farms, with 30% of them testing positive. Serum 

samples were collected from all animals of farms that tested positive to 

ring test. A total of 13,351 serum samples were collected and tested 

with RBT and cELISA assays. Positive index observed with RBT was 

lower when compared to cELISA (0.51 % vs 6.36 %, respectively). 

Furthermore, both tests confirm that positive index in the state of Lara 

is very low. However, relative sensitivity observed in RBT is also very 

low when compared to cELISA. This apparent low sensitivity could 

result in a high number of false negative animals that, consequently, 

will not be eliminated from the herds. Thus, this would constitute a 

constant source of infection to other animals and people. 

Key words: brucellosis, Rose Bengal test, competitive ELISA 

708 In vitro Evaluation of Different Antibiotic Combinations used 

for Treatment of Pasteurellaceae from Bovine Respiratory 

Disease (BRD) 

J. Ganiere 1 , L. Denuault 1 , O. Bidaud 2 , J. Swinkels 3 , F. Payot 2 

1 

Nantes Veterinary School, Research Unit of Infectious Diseases, 

Nantes, France 

2 

Intervet Schering-Plough, Research Unit of Infectious Diseases, 

Angers, France 

3 


Objectives: In the field, cattle with BRD may receive different 

antibiotics simultaneously or successively. The objective of this study 

was to describe the interaction of cefquinome with other antibiotics 

frequently used to treat BRD. 

Materials and Methods: The effect of combinations of cefquinome with 

oxytetracyclin, marbofloxacin, florfenicol, tilmicosin and tulathromycin 

against the Pasteurellaceae involved in BRD: M. haemolytica, P. 

multocida and H. somni was tested in vitro. One strain of each bacterium 

isolated from clinical cases was used to assess the interaction of the 

combinations, determined by the Fractional Inhibitory Concentration 

Index (FICI) and confirmed by time-kill kinetics. Four interactions were 

concerned: FICI

lesions in other organs. Young animals most frequently become infected 

by ingestion of infected milk. We present a case report of tuberculous 

meningoencephalitis in a dairy calf that was less than one year old, who 

presented nervous signs (recumbency, irritability and aggression, 

weakness, dizziness), which progressed to death in three days. At 

necropsy, white nodules were found in the liver, lungs and diaphragm. 

The clinically suspected diagnosis was rabies, but results were negative 

for direct immunofluorescence and intracerebral inoculation in mice. 

Brain samples and spinal cord were preserved in 10% formalin for 

histopathological analysis (HE staining). There was severe granulomatous 

meningomyelitis with Langhans’ giant cells. In cerebral neuropile there 

were some granulomas with a central area of necrosis, perivascular 

cuffing with mononuclear cells, gliosis and hyperemia. The lung had 

many caseated granulomas, a typical finding in bovine tuberculosis, with 

central necrosis, surrounded by epithelioid macrophages, lymphocytes, 

plasma cells, Langhans’ giant cells and fibrosis at the periphery. There 

was mild calcification of the pulmonary granulomas. Samples of lung and 

brain were decontaminated by the Petroff method and inoculated into 

Petragnani and Stonebrink media and M. bovis was isolated. This case 

report shows that bovine tuberculosis should be included in the 

differential diagnosis of bovine neurological disorders. According to 

Brazilian regulation, sanitary measures in the heard infected (test-andslaughter) 

should be adopted. 

FAPESP 

Key words: tuberculosis, Mycobacterium bovis, meningoencephalitis, 

calf 

712 A Study on Diagnosis, Pathogenesis of Pseudomonas 

aeruginosa Isolated from Different Cases of Human and 

Animals in Mosul City - Iraq 

D. Ali 1 , B. Ismaeel 2 

1 College of Veterinary Medicine, University of Mosul, Iraq, 

Department of Microbiology, Mosul, Iraq 

2 College of Veterinary Medicine, University of Dohuk, Department of 

Microbiology, Duhok, Iraq 

The current study tackles the isolation and identification of 

Pseudomonas aeruginosa from different human infection cases 

(wounds, otitis media, urine, stool, and burns) in Al-Zahrawi, Al-Salam 

and Ibn-Al-Atheer hospitals at Mosul city. From animals (sheep, cows, 

goats) samples were also collected (wounds, otitis, urine, feces and 

mastitis) cases for P. aeruginosa isolation and identification. The 

morphological, cultural and biochemical tests confirmed 43 (17.4%) 

isolates as P. aeruginosa from 250 human samples, and 17 (10.3%) 

from 165 animal samples. P. aeruginosa human and animals’ isolates 

virulence factors represented by enzymes (protease, lipease, 

lecithinase, b-lactamase gelatinase, coagulase, haemolysin, toxins) and 

capsule production were studied; 80% of human isolates showed 

capsule presence, while all animal isolates (100%) contained capsule. 

The pigments production especially pyocyanin, which differentiates P. 

aeruginosa from other microorganisms, was detected in 80% of the 

human isolates and in 100% of the animal isolates. The study showed 

similarity between P. aeruginosa of human and animal isolates in their 

virulence factors production ability. The study results revealed that 15 

pathogenic isolates from 43 tested human isolates and 10 pathogenic 

isolates from 17 tested animal isolates produced positive results for an 

exotoxin A production in rabbit skin reaction test, through inducing 

hypersensitivity reactions of type I (early or immediate) and of delayed 

type (Type IV). Baby suckling mice test and rabbit ileal loop test 

confirmed that both human and animal P. aeruginosa isolates produced 

an enterotoxin. The histopathological study revealed different changes 

represented by elongation and blunted villi with necrosis. 

713 Bovine Paralityc Rabies: Prevention and Control in Mexico 

M. Manzano 1,2 , S. Vázquez-Morón 2 , R. Ramirez 1 , M. Orozco 3 , 

J. Echevarría 2 , A. De Vega 1 

1 

Comite de Fomento y Salud Animal, Campana de Rabia, Puebla, 

México, Mexico 

2 

Instituto de Salud Carlos III, Servicio de Microbiologia 

Diagnostica, Madrid, Spain 

3 

Unión Ganadera Regional Sierra Oriente, Gerencia, Teziutlan, 

Mexico 

Bovine paralytic rabies is a contagious and fatal disease that affects a large 

number of productive animal species and also humans, creating a risk both 

to human and animal health. In America this involves a serious problem. 

One of the determining factors to prevent cases of rabies in animals and in 

humans is, without any doubt, the diffusion and training on measures of 

prevention and control of the disease. A DVD on this kind of measures 

was recorded in the City of Puebla, Mexico, by the Committee of 

Promotion and Animal Health and the Foundation “PRODUCE”, Puebla. 

First, the DVD has been presented all over the country, and, on a 

subsequent occasion, it was presented in the RITA (The international 

meeting on research advances in rabies control in the Americas) meetings 

of Brazil and Mexico, in coordination with the Panamerican Health 

Organization. The DVD was distributed to all American countries 

through these meetings.The mentioned DVD shows two aspects of this 

problem: in the first part a case of an outbreak of paralytic rabies is 

narrated in the colloquial language of the cattle farmers from the northern 

mountains belonging to the Puebla State; while the second part includes 

recommendations about the cold chain, antirabies vaccination, sending 

specimens to the laboratory, classification of bats and control of 

hematophagous bats.Furthermore, molecular studies are currently being 

realized in the Instituto de Salud Carlos III (Madrid, Spain) to 

complement the field work made in Mexico. For all the reasons exposed it 

is of great interest to share this experience with European Buiatric groups. 

714 Effect of Cow Vaccination against BRSV and PI3 on Immune 

Status ante Partum and the Transfer of Colostral Antibodies 

to Calves 

B. Makoschey 1 , R. Brunner 2 , M. Koenig 3 , C. Gumpert 4 , H. Heckert 4 

1 


2 

Intervet Schering-Plough, Businessunit Cattle, Unterschleissheim, 

Germany 

3 

Justus-Liebig-Universität Gießen, Faculty of Veterinary Medicine, 

Institute of Virology, Giessen, Germany 

4 

Freie Universität Berlin, Clinic for Ruminants and Pigs, Berlin, 

Germany 

Objectives: The effect of immunization of pregnant cows with 

respiratory pathogens (Bovine Respiratory Syncytial Virus (BRSV) and 

Parainfluenza Type 3 (PI3)) on the antibody levels in the serum and 

colostrum as well the transfer of the colostral antibodies to the newborn 

calves should be determined. 

Material and Methods: The study was performed on a dairy farm where 

the dams and their calves are housed at two different locations. A 

commercial vaccine containing inactivated antigens of BRSV, PI3 and 

Mannheimia haemolytica (M. haemolytica) (Bovipast ® RSP / Bovigrip 

RSP plus, Intervet) was used. Twenty-six dams (n=26) were vaccinated 

twice (6-8 and 2-4 weeks before the anticipated date of calving). A group 

of 19 cows was not vaccinated with the respiratory vaccine. In addition, 

all cows (n=45) were routinely immunized with a neonatal calf diarrhea 

containing inactivated bovine rotavirus, bovine coronavirus and E. coli at 

6 weeks before calving. Colostrum was collected from each cow and fed 

to the respective calf at a volume of at least 4 liter within the first 12 hours 

of life. Blood samples of the dams were taken at the time of the first 

vaccination and around calving. Moreover, samples of the first milkings 

were collected. Blood samples of the calves were taken two days after 

birth. Neutralising antibody titers against BRSV and PI3 were measured 

in the serum and colostrum samples. 

Results: The interpretation of results regarding BRSV was troubled by 

high pre-vaccination antibody levels in a certain proportion of animals. 

Significantly higher antibody titres against BRSV were found in dams 

from the vaccinate group after adjustment of measured titres by 

subtracting individual pre-vaccination values. Moreover, there was a 

correlation between the BRSV antibody titer in the cows and the titers 

measured in the calves. With regard to PI3, vaccination of dams resulted 

in a booster of neutralizing antibodies and a transfer of the antibodies to 

their newborn calves via colostrum regardless the pre-vaccination titer 

against PI3. Moreover, the PI3 antibody titers in the cows correlated well 

with the titers measured in the two days old newborns. 

Conclusions: Cow vaccination against BRSV and PI3 could be a 

strategy to achieve more homogeneous antibody levels in the serum of 

the dams and their calves and can therefore complement the active 

immunisation program in the calves. 

715 Detection of Prevalence of Rotaviral and Corona Viral 

Diarrhea in Newborn Calves 

B. Shojaee 1 , F. Hemmatzadeh 1 , H. Momtaz 1 , M. Ahmadi bozorg 1 , 

B. Khoramian 2 , M. Eslampour 2 


1 Islamic Azad University of Shahrekord, College of Veterinary 

Medicine, Shahrekord, Iran 

2 Facutly of Veterinary Medicine, University of Tehran, Department 


This study performed in order to determine of rotaviral and coronaviral 

diarrhea prevalance in newborn calves in Iran, in the period of 12 months 

(2004-2005). Antigen detection ELISA kit was used for determination of 

rotaviral antigen and Coronaviral antigen in fecal samples. Among 200 

fecal samples 37 of them (18.5 %) were positive to Rotavirus and all of 

them were negative with coronavirus infection. During different seasons, 

the lowest rate of infection was in summer (6%) and high prevalence was 

in winter (48%). Frequency of infection rate of male calves was 14.28 % 

and among female ones was 20.13%. In dairy farms with a proper 

hygienic management, frequency of Rotaviral diarrhea was 5.19% and 

among the calves of the farms without a proper health management this 

frequency was about 63.04%. In different age groups, the least Rotaviral 

frequency was among 1-7 days age group and it was reported to be 3.17% 

and the most one was among the 15-21 days age group with 66.66% of 

infection rate. Statistical analysis of the results showed a significant 

association between infection, season and health management in the farm. 

Infectious frequency among the calves with 15-21 days of ages had a 

significant statistical difference with calves in other age groups. But no 

linear statistical correlation was founded between increase in infection 

rate and age of the calves. 

716 Seroepidemiological Study of BVD, IBR, RSV and PI3 in 

Morocco 

Z. Zouagui, S. Alali, A. Aiyar 



The objective of this study is to assess the seroprevalence of four 

economically important viral infections among Moroccan cattle herds. 

Theses infections are bovine virus diarrhoea (BVD), infectious bovine 

rhinotracheitis (IBR), bovine respiratory syncytial virus (RSV) and 

parainfluenza-3 virus (PI3) infections. Our work aimed also to reveal 

possible relationships between the prevalence of these viruses and 

some epidemiological factors. For this purpose, a large 

epidemiological investigation has been undertaken in six regions, 

known to be important regarding cattle rearing (Doukkala, Haouz, 

Gharb, Moulouya, Tadla and Souss-Massa). 1609 blood samples, 

collected from non vaccinated cattle, have been analyzed by the ELISA 

method to asses their status concerning the four viruses. The 

serological results showed that the general antibody prevalence was 

39%, 23%, 79% and 68% for BVD, IBR, RSV and PI3, respectively. 

However, regional variations of the four infections have been recorded. 

Moreover, the majority of the investigated farms were significantly 

affected by the mentioned viruses. The survey revealed also that cattle 

born and raised in Morocco were more affected than those imported 

since several years. On the other hand, IBR prevalence has been 

partially influenced by the method of service used. 

Key words: BVD, IBR, RSV, PI3, Morocco 

717 Polimerase Chain Reaction Standardization for Semen, Urine 

and Organs of Rams Experimentally Inoculated with B. ovis 

C. Nozaki, N. Lira, J. Megid 

UNESP- School of Veterinary Medicine and Animal Sciences, Higiene 

Veterinária e Saúde Pública, Botucatu-SP, Brazil 

The objective of the study was to adapt and evaluate PCR compared 

with microbiological culture in the detection of B. ovis in semen, 

urine and organs of experimentally inoculated rams. Thirty one rams, 

1-2 years old, were challenged by intrapreputial and conjuntival route 

with a suspension containing 2x109 CFU/mL of B. ovis. All animals 

inoculated were weekly submitted to clinical evaluation, as well as 

collection of blood for serological diagnosis, and collection of semen 

and urine for culture and PCR. From the 31 experimentally inoculated 

animals, PCR detected bacterial DNA in the semen of 24 animals, 

whereas culture showed the agent in 13 rams. PCR showed greater 

sensitivity (77%) than culture (42%). In urine samples, sensitivity of 

the techniques was not significantly different (48% for culture and 

58% for PCR). PCR detected the presence of the agent in 25% of the 

organs tested while culture detected it in only 2% of the samples. 

Although no statistical difference was observed between the 


techniques, results suggest that PCR should be used instead of 

microbiological culture. 

Key words: Brucella ovis, PCR, culture, rams, epididymitis 

718 Serological Profile of Rams Experimentally Inoculated with 

Brucella ovis in the Different Phases of the Illness: 

1 Correlation with Clinical Signals 2 Evaluation of Agar Gel 

Immunodifusion Test and Serum Neutralization Test 

C. Nozaki 1 , N. Lira 1 , O. Augusto Filho 1 , H. Azevedo 2 , L. Rodello 2 , 

S. Bicudo 2 , J. Megid 1 

1 

Universidade Estadual Paulista, Higiene Veterinária e Saúde 

Pública, Botucatu-SP, Brazil 

2 

Universidade Estadual Paulista, Reproduction Animal, Botucatu- 

SP, Brazil 

The present research had as objective to determine the serological 

profile after experimental inoculation of rams with B. ovis using the 

Agar Gel Immunodifusion test and Serum aglutination test (SAR) 

using the commercial kit D-TecCB. The results were correlated to the 

reproductive clinical signals, evaluating the phases of higher 

effectiveness of the serological tests and the method of clinical 

evaluation in the different phases of the experimental illness. Thirty 

one rams, 1-2 years old, were challenged by intrapreputial and 

conjuntival route with a suspension containing 2x10 9 CFU/mL of B. 

ovis REO 198, simultaneously. After inoculation, animals were weekly 

submitted to clinical evaluation and blood collection for serological 

analysis. SAR showed a decreasing percentage of positive results, from 

80% to 11%, from the second to the seventh week after inoculation. 

IDGA showed increasing percentages of positive results, from 45% to 

63%, from the first to the third week, and then a decrease to 4% in the 

11 th week. Fluctuation in titers was observed in both tests. Clinical 

changes began in the first week after inoculation and were associated 

with positive serology in the acute phase of the disease. IDGA 

demonstrated variable sensitivity in the different phases of the disease, 

showing positive animals in the chronic phase, and was preferred in 

routine diagnosis. Negative serological results associated to clinical 

alterations were characteristics of the chronic phase of the disease 

demonstrating the inefficacy of serology in this phase. 

Key words: agar gel immunodifusion test, serum aglutination test, 

sheep brucellosis, clinical signs, serology, experimental inoculation 

719 Seroprevalence of Six Serovars of Leptospira in Human and 

Bovine from Toca, Boyacá (Colombia) 

J. López, M. Pulido, R. Andrade, A. Caycedo 

Universidad Pedagógica y Tecnológica de Colombia, Grupo de 

investigación en medicina veterinaria y zootecnia, Tunja, Colombia 

Objectives: Determining the presence of serovars of Leptospira in 

human and bovine by means of the microscopic agglutination test 

(MAT), and; establishing the probable source of serovars as much in 

the humans as in the bovine ones. 

Materials and methods: Location: six properties milkmaids from the 

rural area of Toca, Boyacá (cold climate). 36 mature people (26 women 

and 11 men) donated 4 ml of blood of the radial vein. 116 samples from 

bovine were collected, corresponding to 8 ml of blood from the caudal 

vein. Samples were centrifuged (3.600 rpm during five minutes) and 

the resulting serum froze to -20° C until its analysis with MAT. 

Results: Serums from people had a positivity of 29.7% (11 of 36); 

being from Canicola 10,8% (4 of 36); Icterohaemorragiae, Pomona and 

Automnalis 5,4% each one, (2 of 36 people each serovar) Hardjo 2,7% 

(1 of 36). The serovar Sejroë was seronegative in human. In bovine 

there was a 48,1% of positivity (64 of 133); being from 

Icterohemorragiae 11,3% (15 of 133), Pomona 13.5% (18 of 133), 

Hardjo 6.0% (8 of 133), Canicola 8.3% (11 of 133), Sejroë 6.8% (1 of 

133); Automnalis 8.3% (11 of 133). 

Conclusions: Prevalence obtained of different servars of Leptospira 

interrogans is adjusted to data obtained by other investigators in 

Colombia. The results don’t offer a precise interpretation about the 

origin of the infection with different stumps of Leptospira interrogans 

so much in human as in bovine. There was not statistical significance in 

any association, in spite of this, infection probabilities exist between 

human and bovine: For serovar icterohaemorragiae 2,22 times in 

bovine that in human; serovar pomona (2,73 times more), hardjo (2,3 

times more) and automnalis (1,57 times more). This would be due to 

the biggest contact among bovine and elements contaminated by urine

or secretions of wild animals, rats or dogs. Serovar canicola seems to 

have a protective effect in bovine (0,74 times with regard to humans), 

being the infection probability by means of secretions of dogs that 

remain in the property. From serovar Sejroë any hypothesis could not 

settle down, because humans went negative to the serovar. Results 

demonstrate the circulation of different serotypes among the animal 

species and they highlight the importance of carrying out more 

advanced epidemic studies, with the purpose of understanding the 

epidemiology of Leptospira in the region. 

Key words: leptospira, human, cattle, Toca, epidemiology 

720 Comparison of Gene Detection by IS6110 and IS1081 

Polymerase Chain Reaction and Isolation by LJ and BM 

Media in Tuberculoid Tissues of Slaughtered Cattle 

YS. Cho 1 , YH. Jean 2 , IS. Roh 2 , I. Hwang 1 , D. Cho 1 , H. Nam 1 , 

E. Heo 1 , JM. Kim1, HR. Han 3 , SC. Jung 1 

1 

National Veterinary Research & Quarantine Service, Bacteriology 

and Parasitology Division, Anyang, Korea, South 

2 

National Veterinary Research & Quarantine Service, Animal 

Disease Diagnostic Center, Anyang, Korea, South 

3 

Seoul National University, College of Veterinary Medicine, Seoul, 

Korea, South 

Objectives of study: Bovine tuberculosis has been diagnosed by the 

examination of tuberculoid tissue as well as tuberculin skin test. Bovine 

tuberculosis at slaughterhouse is diagnosed by histopathological 

finding, polymerase chain reaction, and isolation of Mycobacterium 

bovis. This study provides the isolation rate of Mycobacterium bovis 

from tuberculoid tissues of slaughtered cattle and more efficient 

laboratory methods to diagnose bovine tuberculosis at slaughterhouse. 

Materials and Methods: One part of tuberculoid tissues (n = 89) from 

39 slaughtered cattle were macerated and treated with 0.75% 

hexadecylpyridium chloride as decontaminant. In addition, the other 

part of those were treated with 10% formalin and stained with H&E and 

acid-fast staining kit. The macerated tissues were incubated on LJ w/o 

glycerol and BM media at 37 ˚C for 16 weeks. LJ w/ glycerol media 

were also incubated at 30 ˚C for 16 weeks for the isolation of 

Mycobacterium avium. DNA was extracted by GuSCN-silica and 

IS6110 and IS1081 primers were used for PCR. 

Results: M. bovis was isolated in 82 per cent of slaughtered cattle with 

tuberculoid tissues (32/39). Sensitivity, specificity, positive predictive 

value, and negative predictive value of IS6110 direct PCR (dPCR) 

were 69, 71, 92, and 33, and those of IS1081 dPCR were 79, 71, 79, and 

29, respectively. IS6110 dPCR showed the highest positive predictive 

value, while IS1081 dPCR showed the highest sensitivity. The result by 

dPCR accorded with that by H&E staining, but the tuberculoid tissues 

were not always detected acid-fast bacilli. M. bovis isolation rate by 

BM (77%; 57/74) was higher than that by LJ w/o glycerol (72%; 

63/87). Eighty seven per cent of the isolates (72/83) were IS6110 

positive, and 86 per cent of those (67/78) IS1081 positive. At 30 ˚C on 

LJ w/ glycerol, 16 mycobacteria-like bacilli were isolated from 90 

samples, and 14 isolates of those were identified as Mycobacterium 

bovis by IS6110 and IS1081 PCR. 

Conclusions: The results from IS6110 and IS1081 dPCR were not 

always accorded each other. Therefore, the application of both IS6110 

and IS1081 PCR could increase the diagnostic accuracy of bovine 

tuberculosis. The isolation rate of BM media showed higher than that 

of LJ media. The isolates with only IS6110 or IS1081 gene would be 

further analyzed to examine new genotype of M. bovis in the future. 

Key words: Mycobacterium bovis, bovine tuberculosis, IS6110, 

IS1081, LJ media, BM media 

721 Outbreak of Acute BVD in Brazilian Beef Cattle: 

Clinicopathological Findings and Molecular Characterization 

of a BVDV Strain Subtype 1b 

M. Lunardi, S. Headley, J. Lisboa, R. Otonel, A. Alfieri, A. Alfieri 



When first described, Bovine Viral Diarrhea (BVD) was characterized 

as an acute transmissible disease associated with severe leucopenia, 

high fever, depression, diarrhea, gastrointestinal erosions, and 

hemorrhages. Recently the severe acute form has been related only to 

some hypervirulent BVDV-2 strains. This article reports the detection 

of BVDV-1b associated with an acute and fatal outbreak of BVD in a 

Brazilian herd. Depression, anorexia, watery diarrhea, sialorrhea, and 

weakness were observed in six steers. The affected animals died 24 

hours to 15 days after the first manifestation of infection was observed. 

One of these animals was evaluated for laboratorial, clinical, and 

pathological alterations. Blood samples were collected ante mortem by 

routine procedures. Selected tissues were fixed in 10% neutral buffered 

formalin solution and processed for histopathological evaluation. RNA 

extraction was performed following silica/guanidine isothiocyanate 

method. The RT-PCR assay was carried out using a set of primers 

(103/372) designed from the 5-UTR region of the BVDV genome. The 

identities of the RT-PCR products were confirmed by direct sequencing 

of the amplicon from one clinical specimen. Sequences were examined 

with the software PHRED for quality analysis and the consensus 

sequence was determined using the software CAP3. The alignment was 

obtained by BioEdit software. A neighbour-joining phylogenetic tree 

was constructed using the MEGA program. Laboratory findings were 

non-specific; clinically, the animal was weak, with dehydration and 

erosive oral lesions. Pathological alterations were predominant at the 

tongue, esophagus, and rumen. The RT-PCR assay identified BVDV in 

all clinical samples analyzed. The sequence obtained has been 

deposited in GenBank (accession number EF406123). Phylogenetic 

analysis of BVDV derived from lymph node revealed that this strain 

was clustered with other BVDV subtype 1b isolates, sharing high 

similarity (96.7%) with the representative subtype 1b isolate Osloss. 

The real role played by BVDV-1b in acute diseases and the frequency 

of this subtype in the epizootology of acute and severe BVD are not 

totally elucidated. Molecular characterization is the only definitive 

method to identify the BVDV genotype involved in acute outbreaks of 

severe BVD, since both clinical presentation and pathologic findings 

can be similar to BVDV-2 infection. 


Key words: cattle, BVD, BVDV, 5-UTR 

722 Winter Dysentery by Bovine Coronavirus Infection in a 

Tropical Country 

E. Takiuchi, A. Barry, D. Stipp, R. Otonel, A. Alfieri, A. Alfieri 



Winter Dysentery (WD) is an acute diarrhea caused by Bovine 

Coronavirus (BCoV) that affects adult cattle during the winter with 

high morbidity and low mortality rates. Low temperature and UV light 

intensity during the winter lead to high levels of environmental 

contamination by BCoV. The WD occurrence in dairy herds results in 

economic losses caused by severe reduction in milk production. 

Association between BCoV infection and WD was reported in the 

USA, Europe, Japan and Canada, being sporadic in tropical areas. This 

study reports, the detection of BCoV in an adult cow during an 

outbreak of acute diarrhea in Brazil. The outbreak occurred during 

June, in Ponta Grossa city, Paraná, Brazil and affect 138 from 154 

lactating cows. Three animals died. Most cows had bloody and watery 

diarrhea (three to four days), and milk production dropped from 37 to 

26 liters/cow/day. Fecal sample was collected directly from large 

intestine of a necropsied cow. The sample was negative to C. 

perfringens type C, Eimeria spp, Cryptosporidium spp, enteroxigenic 

E. coli, rotavirus and bovine viral diarrhea virus. Liquid feces 

suspension at 50% (v/v) was prepared in PBS. RNA extraction was 

performed with phenol/chloroform/isoamyl alcohol and 

silica/guanidinium isothiocyanate methods. PCR and semi-nested 

assay were performed according to Takiuchi et al. (2006) with primers 

BCoV1/BCoV2 and BCoV3/BCoV2, respectively, with a predicted 

251 bp product (N gene). The products were analyzed by 

electrophoresis in ethidium bromide stained 2% agarose gel, under UV 

light. BCoV amplicons from semi-nested PCR were purified, 

sequenced, and similarity checked in BLAST software confirming that 

the 251 bp amplicon was from a BCoV strain. This result suggests the 

role of BCoV in the etiology of WD in Brazilian dairy herds. High 

incidence of WD in cold months is notified in regions with well-defined 

seasons. The present case indicates that WD also occurs in tropical 

areas where temperatures are higher. The location of city in the South 

of Brazil, with temperature normally lower, and the season (winter) in 

which the outbreak occurred, perhaps have created favorable 

conditions to virus stability facilitating dissemination among confined 

cattle. In conclusion, while there are some studies associating BCoV 

with outbreaks of WD in other countries, there are a few reports in 

South America, Brazil. 



Key words: dairy herd, winter dysentery, Bovine cCoronavirus 

723 Frequency of BCoV in Neonatal Calf Diarrhea of Brazilian 

Herds 

D. Stipp, A. Barry, E. Takiuchi, R. Otonel, A. Alfieri, A. Alfieri 



Neonatal diarrhea is one of the main causes of calf morbidity and 

mortality that determine economical losses to cattle herds worldwide. 

Several environmental, managemental, nutritional and physiological 

factors may occur either alone or in synergy with the different 

infectious agents. The rotavirus is considered the main etiological 

agent of viral gastroenteritis in mammals. However, other viruses have 

been identified in bovine neonatal enteric infections such as the Bovine 

Coronavirus (BCoV). Epidemiological data on the frequency of BCoV 

occurring in South America livestock farms is very limited. Besides, 

the applicability of a sensitive and specific diagnostic technique such as 

RT-PCR is fundamental to not underestimating the real prevalence of 

BCoV. We evaluate the BCoV frequency in diarrheic fecal samples 

from calves up to 60 days old from beef and dairy cattle herds by a semi 

nested-PCR assay (SN-PCR). The animals were from four Brazilian 

States: Sao Paulo (SP), Paraná (PR), Minas Gerais (MG) and Mato 

Grosso (MT). The samples were prepared as either as 10% (w/v) 

suspension of semi-solid feces or as a 50% (v/v) suspension of liquid 

feces in PBS pH 7.2. For RNA extraction a combination of phenol/ 

chloroform/ isoamyl alcohol and silica/isothiocyanate methods was 

performed. The primers were designed from the highly conserved 

region of the nucleocapside gene of the Mebus strain. For the first 

round of amplification the primers BCoV1 and BCoV2 were used and 

designed to amplify a fragment of 454 bp. For the second round the 

primers BCoV3 and BCoV2 were used that was expected to amplify a 

product of 251 bp. Among the 262 diarrheic fecal samples included in 

this study, BCoV was detected in 15.6% (41/262) of the samples of 

which 37.5% (6/16), 11.1% (6/54), 12.2% (22/181) and 63.6% (7/11) 

were from SP, PR, MT and MG, respectively. The BCoV infection was 

more frequent (24.1%) in diarrheic fecal samples from calves between 

the third and fourth weeks of life. The rate of positive results confirms 

that the BCoV is an important etiological agent of the acute neonatal 

diarrhea in Brazilian dairy and beef cattle herds. The SN-PCR used in 

this study demonstrated to be an accurate tool for the routine detection 

and diagnosis of BCoV in feces from naturally infected calves. 


Key words: dairy herd, winter dysentery, Bovine Coronavirus 

724 Assessment of BoHV-5 A663 Growth Kinetic and 

Comparison with BoHV Reference Strains 

M. Ladelfa 1 , F. Kotsias 1 , M. Del Medico Zajac 2 , S. Romera 2 

1 

CONICET, Buenos Aires, Argentina 

2 

Instituto Nacional de Tecnología Agropecuaria-INTA, Buenos Aires, 

Argentina 

Bovine Herpesviruses 1 and 5 (BoHV-1, BoHV-5) are two closely 

related alphaherpesvirus which co-circulate in Argentina. BoHV-5 

causes fatal encephalitis while BoHV-1 is associated with abortions, 

respiratory and genital disease. BoHV-5 strains are divided into three 

subtypes. a (N569 strain), b (A663 strain) and non a/b. A663 was 

isolated in our country and is considered a reference strain. The aim 

of this work is to establish the growth kinetic of the BoHV-5 A663 

and compare it with BoHV-5 N569 and BoHV-1 LA strains. For that 

purpose, one step and multiple steps kinetics were performed. MDBK 

cells were infected with the strains mentioned above at a MOI of 0.1 

and 5, virus adsorption was synchronized and then incubated at 37˚C. 

After that, extracellular virus was inactivated. At 0, 3, 6, 12, 24, 36 

and 48 hours post infection (hpi) and 0, 3, 6, 12, 15, 18 and 24 hpi, for 

MOI 0.1 and 5, respectively, intra and extracellular fractions were 

obtained, as well as total fraction, and titrated twice on MDBK in 

duplicate.The growth kinetics curves showed similar profile among 

LA, N569 and A663 strains. Virus titre reached at the end of the assay 

was 10 6.7 , 10 6.8 and 10 16.7 TCID50/ml at MOI 5 and 10 7.4 , 107 and 

10 7.2 TCID50/ml at MOI 0.1 for LA, N569 and A663, respectively. 

Maximum viral titre was obtained at 18, 24 and 12 hpi at MOI 5 and 

48, 36 and 48 hpi at MOI 0.1 for LA, N569 and A663, respectively. 

The A663 extracellular fraction showed a remarkable difference at 


MOI 0.1 in comparison with LA and N569, rising lower titres at all 

the times assayed. Virus release percentage was similar among the 

strains at both MOI and the maximum figures were in all cases near 

to 50 %. Finally, the eclipse period was analised. At MOI 5, it 

finished at 3, 6 and 12 hpi for LA, A663 and N569 respectively, and 

intracellular virus was detected at 3 hpi in all cases. At MOI 0.1, the 

eclipse period finished at 12 hpi for all three viruses. Intracellular 

virus was detected at 3 hpi for LA and at 6 hpi for A663 and N569. 

These results provide relevant information about virological aspects 

of BoHV-5 A663 strain, for which there is scarce information 

available, and allow its comparison with other related viruses. 

Concerning this, although A663 and N569 growth kinetics are 

similar, significant differences in the extracellular fraction titres were 

observed at low MOI, while LA strain behaves like N569. Besides, 

these results constitute an important tool for future in vitro 

experiment design. 

Key words: BoHV-5 A663, BoHV-5 N569, BoHV-1 LA, growth 

kinetic 

725 A BoHV-1 gE Gene Deleted Live Vaccine is not Excreted and 

does not Establish a Latent Infection in Trigeminal Ganglia 

after Infection in Calves 

F. Kotsias 1 , M. Del Médico Zajac 2 , M. Ladelfa 1 , S. Romera 2 

1 

CONICET, Buenos Aires, Argentina 

2 

Instituto Nacional de Tecnología Agropecuaria-INTA, Buenos Aires, 

Argentina 

A recombinant Bovine Herpesvirus 1 (BoHV-1) strain lacking the 

glycoprotein E gene (gE gene) (BoHV-1ƒgE‚gal) has been previously 

developed in our laboratory. Although its protective, immunogenic and 

safety properties have already been assessed when used as a live vaccine, 

some features concerning its pathogenesis, different from those reported 

for other gE gene deleted BoHV-1 strains, remained unstudied. In order 

to characterize the BoHV-1ƒgE‚gal strain in its natural host so as to 

understand its unique biological behavior, we now present the 

preliminary results concerning in vivo properties of this live vaccine 

candidate virus. Six calves were housed in an isolation stable, and 

intranasally inoculated with a high dose of the BoHV-1ƒgE‚gal virus. 

Animals were monitored until days 9, 15, 21 and 42 post-infection (pi) 

when they were euthanized. The two remaining calves were treated with 

dexametasone in order to reactivate latent virus, and two weeks after they 

were euthananized at 65 and 87 days pi. Nasal samples were tested for 

infectious virus, viral DNA and IgA antibodies while IgG antibodies 

were evaluated in serum samples. Trigeminal ganglia (TG) tissue 

samples were examined for viral DNA. Animals showed no clinical 

illness and rectal temperature remained normal throughout the 

experience. Infectious virus could not be isolated from nasal swabs at any 

day pi, however viral DNA was detected by PCR at days 4 and 7 pi. 

Serum IgG antibodies were found in all animals except for calf 

euthanized on day 9 pi. Moreover, presence of IgA antibodies in nasal 

secretions were detected on days 7, 9 and 12 pi in all six calves. After 

dexametasone treatment, no infectious virus could be detected in nasal 

swabs nor clinical manifestations observed. PCR from TG were negative 

for all the animals under study. The BoHV-1ƒgE‚gal strain used in this 

experience proved to be avirulent in calves, since no clinical signs were 

observed nor virus shedding detected. Results from serum antibodies 

taken together with the presence of IgA in nasal secretions indicate that 

calves elicited a detectable immune response. Absence of viral DNA in 

all TG and failure to reactivate both infected animals suggest that this 

recombinant virus does not establish a biologically important latent 

infection. This is the first report on a gE gene deleted BoHV-1 virus not 

excreted after infection nor detected in TG. For all this, BoHV-1ƒgE‚gal 

strain constitutes an epidemiologically safe vaccine candidate. 

Key words: BoHV-1, gE gene deleted, live vaccine, cattle, latency 

726 Serological Survey of Coxiella burnetii Infection in Herds of 

Caltanissetta District 

A. Stancanelli 1 , D. Macr 2 , C. Di Bella 2 , A. Torina 2 , S. Agnello 1 

1 IZS Sicilia, Area Caltanissetta, Caltanissetta, Italy 

2 IZS Sicilia, Palermo, Italy 

Q fever is a worldwide zoonosis caused by a gram-negative obligate 

intracellular bacterium, Coxiella burnetii. Although Q fever is present 

virtually in all “animal kingdoms”, including arthropods, the disease 

affects mostly humans, cattle, sheep and goats. The aim of this study

was to evaluate, by serology, prevalence of Coxiella burnetii infection 

in herds of Caltanissetta district. 124 of 289 herds were selected, with 

stratified random sampling, distributed in four parts of Caltanissetta 

district. Particularly 71 herds were chosen from Caltanissetta territory, 

9 from Gela, 29 from Mussomeli and 15 from San Cataldo. 1711 sera 

were collected to test the presence of specific antibodies to C. burnetii. 

Sera were tested by a commercial available ELISA (Chekit Q-fever; 

IDEXX Laboratoires). Results showed serological positivity in 60/124 

(48,4%) herds with 118 positive sera/1711 (6,9%). 

727 Apparent Prevalence of Antibodies to Coxiella burnetii (Q 

Fever) in Bulk Tank Milk of Dairy Herds in the Walloon 

Region of Belgium 

G. Czaplicki 1 , J. Houtain 1 , C. Manteca 2 , C. Saegerman 3 

1 

ARSIA, Diagnostic Center for Animal Diseases, Loncin, Belgium 

2 

CEVA santé Animale, Diagnostic Center for Animal Diseases, 

Libourne, France 

3 

University of Liege - Faculty of Veterinary Medicine, Epidemiology 

and Risk Analysis applied to Veterinary sciences, Faculty of, Liege, 

Belgium 

Objectives: Q Fever is a zoonotic disease caused by Coxiella burnetii, 

an obligate intracellular bacteria. Ruminants (sheep, goats and cattle) 

remain the major natural reservoir for human infection. The aim of the 

present study was on the one hand, to evaluate the apparent prevalence 

in the Walloon dairy herds and on the other hand, to identify herd level 

risk factors and clinical signs statistically associated with seropositivity 

in the population of randomly selected Walloon dairy cattle. 

Material and methods: 566 herds were randomly selected according to a 

classical methodology (Jenicek and Cléroux, 1987). Herd level predictors 

data were collected from a questionnaire on farm demographics, 

management practices and observed clinical signs during the twelve last 

months. Two hundred and six farms responded on a voluntary basis to this 

questionnaire and submitted a sample of bulk tank milk sampled in 

February 2006. Bulk milks were centrifuged and stored at 20°C until 

testing for antibodies to Coxiella burnetii with a commercial Indirect Elisa 

test (LSI). Apparent prevalence was estimated with a ninety-five percent 

confidence intervals (95% CI) assuming a binomial exact distribution. 

Statistical analysis of the data was done using a chi square test and the 

tendency of each parameter to become a risk factor was evaluated by an 

odds ratio calculation with 95% confidence intervals (logarithmic 

approximation); a P value < 0.05 was considered significant. 

Results: Global apparent prevalence: 54.9 % (95% CI : 47.8-61.8%) of 

herds show serological evidence of Coxiella burnetii infection. Clinical 

signs as abortion in heifers and in cows, stillborn or weak calves are 

statistically associated with seropositivity. Risk factors associated with 

seropositivity are free stalling and watering with well water while 

blocked stalling, watering with tap water and disinfection of sheds act 

as protectors of infection. 

Conclusions: This first report on Q Fever infection prevalence in 

Walloon dairy herds indicates a quite high level of infection. Seropositivity 

at herd level does not mean shedding of Coxiella burnetii, but 

identifies herds where at least 10% of lactating animals are sero-positive 

to C. burnetii. A significant association exists between sero-positivity 

and typical clinical signs of Q Fever observed in those herds. Some risk 

factors and some protectors are identified on a statistical basis. 

Key words: bovine, Q Fever, serology, prevalence 

728 Identification of Bovine Viral Diarrhoea Virus 

Mimotopes/Epitopes through Selection from a Peptide 12-mer 

Phage Display Library 

A. Zamit 1 , M. Ostrowski 2 , N. Fondevila 1 , O. Lopez 3 , A. Bratanich 4 , 

S. Romera 1 

1 

Instituto Nacional de Tecnología Agropecuaria-INTA, BUENOS 

AIRES, Argentina 

2 

Institut Curie, Paris, France 

3 

Northern Michigan University, Marquette, United States 

4 

Facultad de Cs Veterinarias-UBA, BUENOS AIRES, Argentina 

Bovine Viral Diarrhoea Virus (BVDV) is a worldwide distributed 

pestivirus that causes important economical losses to cattle industry. 

Argentine is no exception with a population of 50 million bovines and 

high seroprevalences up to 90%. The aim of our study is epitope 

mapping of immunodominant E2 protein of BVDV using a peptide 12mer 

phage display library. 

Materials and Methods: A peptide 12-mer phage display commercial 

library was depleted of mimotopes resembling BVDV proteins except 

those corresponding to E2, by using serum from a calf immunized 

against recombinant BVDV-E2 BDV. Then, a positive selection was 

performed with affinity purified antibodies against BVDV strain NADL. 

Individual phage clones were amplified and displayed peptide sequences 

analyzed using CLUSTALX with Tudos matrix. Four peptides were 

designed for immunization purposes and sent to synthesize. Two bovines 

and 4 groups of guinea pigs were vaccinated with peptides coupled to 

KLH. Also, phages displaying chosen peptides were amplified and used 

to immunize mice. Sera were analyzed by peptide-ELISA, phage- 

ELISA, indirect ELISA and seroneutralization. 

Results: Enrichment of specific clones was achieved in 4 rounds of panning 

and 30 clones were individually amplified. Multiple sequence alignment 

analysis with modified parameters and substitution matrix allowed a better 

clustering of peptides. Considering clusters, peptide frequency and 

hydrophobicity and aminoacids representativity in library, 4 rationally 

designed peptides with immunogenic properties were obtained. Satisfactory 

antibody responses against peptides were observed in mice immunized with 

phages, and bovines and guinea pigs vaccinated with synthetic peptides. 

Also, antibodies produced in mice recognized synthetic peptides. Sera from 

immunized animals were either citotoxic or not neutralizing. Indirect 

ELISA showed high background and was not conclusive. 

Conclusions: It was possible to obtain immunogenic peptides 

resembling epitopes of BVDV using a peptide 12-mer phage display 

library as mapping strategy. Epitope mapping is imperative for 

improving local vaccines design and control. 

Key words: BVDV, mimotope, phage display, E2 

729 Amplification of Inv Gene of Salmonalla Serotypes by 

Polymerase Chain Reaction (PCR) as a Specific Method of 

Detection of Salmonellae 

T. Zahraei Salehi 1 , M. Mahzonieh 2 , A. Ashrafi 1 

1 

Faculty of Veterinary Medicine, University of Tehran, Microbiology, 

Tehran, Iran 

2 

Faculty of Veterinary, Pathobiology, Shahrekord, Iran 

Background: Detection of inv gene in Salmonella serotypes by PCR. 

Materials and Methods: Sixty Salmonella strains were isolated from 

animals and human sources. In this research, at first 60 isolated 

Salmonella from animals and human were tested by biochemical tests 

such as carbohydrate utilization tests and urease test and then were 

serogrouped by Salmonella O anisera. DNA of isolated Salmonella 

were extracted by Holmes and Quigley method. Two primers (St139 

and St141) and PCR reagents from fermantas company were used for 

amplication of inv gene. PCR reaction was carried out in Master cycle 

(Eppendorf). The PCR product were loaded into 1.2% agarose gel and 

electrophoresed for 60 minutes at 120 V. 

Results: All isolates showed biochemical properties of Salmonellae. In 

PCR assay, target gene (invA gene) with 284 bp size were observed in all 

the strains, which is corresponded with size of band of positive control and 

DNA marker. So, all of the strains in this survey had invA gene. 

Conclution: According to the results of this study PCR method based 

on inv gene is useful to rapid identification of Salmonella serotypes. 

Key words: Salmonella, invA gene, PCR 

730 Toxoplasmosis in a Commercial Dairy Farm in Turkey 

S. Gazyagci 1 , C. Babur 2 , S. Kilic 2 , B. Celebi 2 , A. Gazyagci 2 , 

B. Yagci 1 

1 

Faculty of Veterinary in Kirikkale University, Department of 

Internal Medicine, Kirikkale, Turkey 

2 

National Research Center, Epidemiology and Public Health, 

Ankara, Turkey 

Blood samples were taken from 742 dairy cattles in a commercial farm. 

Antibody titers were determined against Toxoplasma gondii by Sabine- 

Feldman Dye method. Serological examination revealed 640 cattle to 

be seropositive. Among these sera the numbers of seropositive samples 

at certain dilution steps were as follows: 268 at 1/16, 184 at 1/64, 112 

at 1/256 and 76 at 1/1024. Three cats could circulate in the warehouse 

in the farm. All sera were tested with Sabin-Feldman Dye test for 

Toxoplasma gondii spesific antibodies. Antibody titers were 1/256 all 

of them.This study investigates the seroprevalence of Toxoplasma 

gondii in dairy cattle from a commercial farm in Turkey. 

Key words: cattle, Sabin Feldman dye test, Toxoplasma gondii 


731 Detection and Investigation on the within-farm Transmission 

of Bovine Herpesvirus-1 in Farms Located in the Trace 

District of Turkey 

A. Unver 1 , N. Turan 2 , H. Yilmaz 2 

1 Farmamag, Firuzkoy Yolu, Baglar Sokak, Avcilar, Istanbul, Turkey 

2 University of Istanbul, Veterinary Faculty, Department of Virology, 

Avcilar, Istanbul, Turkey 

The aim of this study was to detect BHV-1 infection and to investigate 

within-farm transmission in farms located in the Trace district of 

Turkey. For this, 3 dairy farms having the animals over 20 were 

selected from the Marmara region. Two visits were made to these farms 

6-7 months intervals. At each visit, the nasal swabs and blood from 

each animal and the milk sample from milking cows were taken. Blood 

sera were analysed by ELISA (IDEXX, gB blocking) for the presence 

of antibodies to BHV-1. Whereas the PCR was used to detect BHV-1 

DNA in the nasal swabs and milk samples. In samples taken during the 

first visit, antibodies to BHV-1 were detected in 13 of 89 animals in 3 

farms. 4 of 27 (14.8%) animals, 6 of 21 (28.7%) animals and 3 of 41 

(7.3%) animals were found to be seropositive in farms-1, 2 and 3, 

respectively. BHV-1 DNA was detected in nasal swabs of 2 animals 

and milk sample of one animal in farm-1, milk sample of 1 animal in 

farm-2 and nasal swabs of 6 animals in farm-3. In samples taken during 

the second visit, antibodies to BHV-1 were detected in 18 of 71 animals 

in 3 farms. 6 of 24 (25%) animals, 6 of 17 (35.2%) animals and 6 of 30 

(20%) animals were found to be seropositive in farms-1, 2 and 3, 

respectively. BHV-1 DNA was detected in nasal swabs of 3 animals in 

farm-1, nasal swabs of 2 animals and milk sample of 1 animal in farm- 

2. No BHV-1 DNA was detected in the samples taken from the farm-3. 

In conclusion, the BHV-1 infection was detected by ELISA and PCR in 

all 3 farms. Some of the animals found to be seronegative by ELISA 

were positive on PCR and these animals might be either seronegative 

carriers or acutely infected. Therefore, it would be better to check the 

animals in the farms by both test ELISA and PCR. Also care must be 

taken to test new animals and separating the youngs from the adults. 

Key words: bovine, herpesvirus-1, transmission, PCR, ELISA, Turkey 

733 Outbreak of Foot-and-Mouth Disease with High Fatality Rate 

in Young Lambs in Tabriz - Iran 

G. Noursaadat 1 , M. Tooloei 2 , F. Rezazadeh 2 

1 Large Animal Clinician, Tabriz, Iran 

2 University of Tabriz, Faculty of Veterinary Medicine, Department of 

Clinical Science, Tabriz, Iran 

Foot-and-mouth disease (FMD) is caused by an aphthovirus (family 

picornaviridae) and affects all cloven foot animals. It is endemic in 

Africa, Asia, South America and parts of Europe. The disease in adult 

sheep usually causes milder clinical signs than in cattle or pigs, and is 

often subtle enough to go undiagnosed. In contrast, FMD in lambs has 

been reported to cause high mortality during field outbreaks. From 

January to April 2007, five to fifty days-old lambs of the approximately 

all of the flocks pertaining to the east region of Tabriz affected with a 

highly fatal disease. The morbidity and mortality rates in lambs of each 

flock in this outbreak were about 10-50 and 70-80%, respectively. 

Adult sheep had also been affected, but the disease in sheep was very 

mild and with less frequent and extent than lambs. All affected lambs 

were clinically examined. Observed clinical signs include: severe 

tachycardia, high fever (up to 42˚C), tachypnea, dispnea, salivation and 

drooling, development of vesicular lesions in interdigital and coronary 

region of all four foot and in mouth especially over the tongue and lips 

as well as lameness. Most of the affected sheep were also suffering 

from lameness. At the necropsy of at least one hundred died lambs, 

hyperemia and hepatization of lungs especially in the apical and ventral 

lobes, swelling and enlargement of mediastinal lymph nodes, 

petechiation of spleen and pericardial fats, pericarditis and pericardial 

effusion, epicardial and myocardial hemorrhages with pale and 

necrotic areas or the typical tiger heart appearance and hepatomegaly 

were observed. Histopathologic examination revealed severe hyaline 

degeneration and necrosis of myocardial fibers and marked interstitial 

infiltration by mononuclear cells as well as congestion and periacinar 

hepatocellular necrosis in the liver. The diagnosis of Foot-and-mouth 

disease was confirmed by the evaluation of the clinical and necropsy 

findings as well as histopathologic changes. Mortality rates in the 

affected flocks mainly decreased through vaccination of the all affected 

and unaffected lambs and sheep even one-day old lambs and also 


administration of some antibiotics such as Trimetoprim, Sulfadiazine 

and Gentamycin for 72 hours. 

Key words: foot-and-mouth disease, lambs, Tabriz 

734 Macroscopic and Microscopic Studies on Pathological Lesions 

of Deleted Ovine Livers in Kermanshah Slaughterhouse 

Dr. Bahiraie 1 , Dr. Pouyanmehr 1 , M. Razmju 2 

1 

Razi University,Faculty of veterinary Medicine, Department of 

Animal Health and Anatomy, Kermanshah, Iran 

2 

Faculty of Veterinary Medicine, Meat Hygiene Department, Razi 

University 

Objective study: The pathological survey on liver which had been 

recorded from sheep carcasses in slaughterhouse of Kermanshah. The 

indication of lesions which causes liver deletion during slaughtering 

process. 

Material and Methods: The macroscopic and microscopic 

examination carried out on 160 sheep livers which were recorded in 

city slaughterhouse. The samples were collected randomly during 1 

year (2006). Macroscopic survey was done on each recorded liver. 

Preparation for microscopic examinations carried out on macroscopic 

lesions samples by use of routine histological methods. Dimension of 

samples were 5x, 5x, 5x millimeters. The technique for preparation of 

tissue sections by tissue processor (Histokinettte) equipment was 

including: 1-Fixation (buffere formalin 10%, 2-3 days), Washing 

(water 4-5 hours), dehydration (using of ascending concentrations of 

ethylic alcohols70%, 80%, 90%, 100%, 100%), Clearing(Xylene), 

Impregnation (paraffin 56 ˚C-68 ˚C), Blocking (paraffin Merck), 

Sectioning (Microtome).The thickness of prepared sections for study 

was 5. Staining was Haematoxylin & Eosin (H&E). Statistical analysis 

was done by SPSS. 

Results: The highest macroscopic lesion was cirrhosis and then lesions 

due to hydatid cysts and Fascioulosis. In microscopic study, the highest 

diagnosed lesions was Bacterial hepatitis 20 cases (12.5 %) and then 

pathological lesions due to Fascioulosis 18 cases (11.35%), hepatic 

cirrhosis16 cases(%10), Chronic bill ducts inflammation 16 cases(10%), 

Telangiectasia16 cases (10%), Dicroselliousis 14 cases (8.75%), Chronic 

hepatitis 14 cases (8.75%), Hydatid cyst 10 cases (6.25%) , Fatty Liver 8 

cases (5%),Hemorrhagic Pies hepatitis 8 cases (5%), Blood congestion in 

sinusoidal perihepatitis and Eosinophilic parasitical hepatitis each of 

them 6 cases (3.75%), Cholangitis and fibroma 4 cases (2.5%). In this 

research, centrilobular Necrosis and cholangiocarsinoma which reported 

in another spices was none found (0.0%). 

Conclusion: The effects of contamination to parasitical agents 

identified as the most causes of deletion of sheep liver directly or 

indirectly. Even thought the cirrhosis was major factor in macroscopic 

judgment for deletion, but the parasitical agents are the essential factors 

for deletion of lives. 

Key words: sheep health, meat hygiene, pathological lesions of liver, 

judgment of carcasses 

736 Antibacterial Activity of Aqueous Methanol Extracts of 

Zingiber Officianale, Curcuma Longa, Vernonia 

Anthelmintica, Acacia Nilotica and Melia Azedarach 

Z. Abbas 1 , M. Aslam 2 , M. Arshad 2 , Z. Iqba 1 , M. Ashraf 2 

1 

University of Agriculture, Department of Parasitology, Faisalabad- 

38040, Pakistan 

2 

University of Agriculture, Department of Microbiology, Faisalabad, 

Pakistan 

Introduction: The development of drug resistance as well as 

appearance of undesirable side effects of certain antibiotics has led to 

the search of new antibacterial agents in particular from medicinal 

plants. Therefore, the present study was conducted to assess the 

antibacterial potential of aqueous methanol extract of Zingiber 

officianale, Curcuma longa, Vernonia anthelmintica, Acacia nilotica 

and Melia azedarach against Staphylococcus aureus, Escherichia coli 

and Bacillus subtilis. 

Materials and Methods: The antibacterial activity of these extracts 

was tested using disc diffusion method. Filter paper discs were soaked 

with 15ul of plant extracts stock solution and were allowed to dry at 

room temperature for 10-15 min. A bacterial culture was uniformly 

spread on the surface of Mueller Hinton agar plates using a sterile 

swab. Each disc was pressed down uniformly against the surface of 

agar and then examined for any inhibition zone if present. The test was

epeated three times to ensure reliability.The MIC was also determined 

by using agar dilution method and agar well diffusion method. 

Results: Aqueous methnol extract of Acacia nilotica was found 

effective against Staphylococcus aureus and Bacillus subtilis. The 

other four extracts did not show any inhibitory effect against any of the 

three bacterial species. The Minimum Inhibitory Concentration (MIC) 

values of aqueous methanol extract of Acacia nilotica determined by 

agar dilution method and agar well diffusion method against 

Staphylococcus aureus were 0.78 mg/ml and 1.56 mg/ml, respectively. 

In case of Bacillus subtilis, MIC value of extract of Acacia nilotica as 

determined by agar dilution method and agar well dilution method 

were 1.56 mg/ml and 3.125 mg/ml, respectively. 

17 Effect of Vaccination against Enterotoxaemia of Pregnant 

Ewes on Antitoxin Alpha and Epsilon Antibodies in 

Colostrum, Milk and Lambs after Colostrum Intake 

M. Socié-Jacob 1 , B. Bolkaerts 1 , L. Wiggers 1 , J. Van de Ven 2 , 

N. Kirschvink 1 

1 

University of Namur, Veterinary Department, Animal Physiology, 

Namur, Belgium 

2 

Schering-Plough, Animal Health, Brussels, Belgium 

Enterotoxaemia caused by C. perfringens belongs to the pathologies 

in lambs that often causes mortality. Protection against 

enterotoxaemia can be provided by vaccination, but the development 

of a specific immune response by newborn lambs lasts 4-6 wk. The 

aim of this study was to test whether vaccination of ewes against 

enterotoxaemia would provide specific colostral- and milkassociated 

anti-toxin and antibodies to suckling lambs. Ewes were 

divided in a unvaccinated control group (C:n=90) and in a 

vaccination group (V:n=130) that underwent vaccination with a 10valence 

vaccine 6 (T-6) and 2 wk prior to lambing. Blood of ewes 

(C:n=36;V:n=53) was sampled before vaccination and at lambing 

(T0). Colostrum was sampled at lambing (C:n=90;V:n=130) and 

blood was sampled in lambs 24h after birth (C:n=60;V:n=90). 

Ewes’milk (C:n=60;V:n=110) was sampled at weaning (9-12 wks 

after lambing). Samples were analysed for antitoxin and antibody 

(Ab) levels by use of an EIA. 

Results: Data are shown as mean SE and are expressed in arbitrary 

units of Ab. 

Ewes Ewes Colostrum Milk at Lambs 

serum serum T0 weaning serum 

T-6 T0 T24 

C (Ab) 20 12 25 19 22 16 12 5 47 6 

(Ab) 7 13 8 11 9 15 4 4 28 58 

V (Ab) 51 12 887 883* 583 49* 83 23* 2935 603* A 

(Ab) 37 11 1721 2929* 2075 305* 50 10* 10008 2381* A 

*Significantly different from respective C-value, P

necropsied, tissue samples were taken and processed for pathologic 

examinations. Blood samples of 15 ewes (dams of the 20 dead lambs) and 

3 rams were collected and then Sabin Feldman dye test was used for the 

detection of T. gondii spesific antibodies and respectively 13 (86,6%) and 

2 (66,6%) were found to be positive. Toxoplasmosis in pregnant ewes 

generally results with abortion by the way either following a new acute 

infection or reactivation of the chronic infection during pregnancy. 

Therefore, abortion can be used as an indicative of a transmitted infectious 

disease from dam to fetus. So it may be has a key role on differential 

diagnosis of such a herd problem e.g. toxoplasmosis. Postnatal lamb 

deaths and lacking abortion during pregnancy are not frequently seen in 

natural toxoplasmosis in sheep. However, death of a large number of 

newborn lambs in a herd should be investigated fortoxoplasmosis, 

serologically and etiologically. 

Key words: Toxoplasma gondii, Sabin Feldman dye test, 

immunohistochemistry, fatal toxoplasmosis, lamb 

21 The Role of Estradiol and Somatotropin in the Pathogenesis of 

Ovine Pregnancy Ketosis 

R. Everts 1,2 , R. Jorritsma 1 , M. Houweling 2 , L. Leengoed Van 1 , 

A. Tielens 2 

1 

Utrecht University, Department of Farm Animal Health, Utrecht, 


2 

Utrecht University, Department of Biochemistry and Cell Biology, 

Utrecht, Netherlands 

Ovine pregnancy ketosis (pregnancy toxemia, twin lamb disease) is the 

most common metabolic disorder of ewes during the last weeks of 

pregnancy. Ewes with the disease normally carry two or more lambs. It 

is biochemically and endocrinologically characterized by 

hypoinsulinemia, ketosis, and an increase in the plasma concentrations 

of non-esterified fatty acids (NEFAs), cortisol, estrogens and 

somatotropin. While morbidity is low, the reported mortality in 

absence of therapy is 90%. It was thought that the main cause of 

pregnancy ketosis in sheep is a disturbance of carbohydrate metabolism 

by the high demands for glucose by de developing fetuses in the last 

trimester of pregnancy. For several reasons the appropriateness of this 

hypothesis can be questioned. First, the glucose turnover rate during 

lactation is two to three times the demand during pregnancy, without 

according increase in ketone bodies during lactation. Second, fasting of 

pregnant ewes does not induce all clinical and endocrinological aspects 

of pregnancy ketosis. In contrast to ewes with spontaneous pregnancy 

ketosis, ewes with induced ketosis start eating almost immediately 

upon refeeding. Third, the observation that immediately after (induced) 

delivery, a rapid recovery of the ewe occurs despite the increased 

energy demand for the production of milk, further questions the 

importance of a high demand for glucose as the sole inducer of ovine 

pregnancy ketosis. Therefore, we propose that the rise in 17‚-estradiol 

and somatotropin, seen in affected ewes, hampers the normal metabolic 

regulation of the negative energy balance, resulting in pregnancy 

ketosis. The aim of this study was to investigate the influence of 

estradiol and somatotropin on the lipid metabolism in isolated sheep fat 

cells and to compare the lipid metabolism in sheep with pregnancy 

ketosis to that in healthy pregnant sheep. For that reason, subcutaneous 

fat biopsies were taken from sheep at the perineal area of the tail base. 

Lipolysis was assessed in freshly isolated fat cells and after 30h of preincubation. 

Furthermore, in the fat biopsies mRNA levels of several 

metabolic regulators such as PPAR, adiponectin and leptin were 

assayed. 

Key words: ketosis, ovine, lipid metabolism, estradiol, somatotropin 

22 Individual Animal Risk Factors for Ovine Clinical Mastitis 

S. Waage 1 , S. Vatn 2 

1 Norwegian School of Veterinary Science, Production Animal 

Clinical Sciences, Oslo, Norway 

2 Animalia, Oslo, Norway 

Clinical mastitis (CM) occurs frequently in ewes, causing great 

economic loss and a serious animal welfare problem in the sheep 

industry. 

The objectives of this study: were to examine the distribution of ovine 

CM cases in relation to the time of lambing and identify and quantify 

the effect of risk factors for the disease. 

Materials and methods: We conducted a case-control study, using 

individual animal data and disease records from a national sheep 


registry in Norway. Ewes that lambed in the spring of 2004 were 

included. Eligible cases (n=2857) and controls (n=76,716) from 1056 

flocks of meat sheep were matched on flock (m:n matching) and 

multivariate conditional logistic regression was used for 

analysis of the data. 

Results: A large proportion of the CM cases occurred close to parturition; 

32% were treated between days 0 and 4 after lambing, and 55% were 

treated within 2 weeks postpartum. Increasing age of the ewe and the need 

for assistance at lambing owing to dystocia were associated with 

increased CM risk. However, the effects of both these factors were 

modified by the number of lambs born and were most pronounced for 

ewes with 1 lamb. Increasing number of lambs born to the ewe, including 

stillborn, was associated with a marked increase in the odds of CM. For 

example, odds for 2-year old ewes without dystocia were 6.7 times greater 

for those with more than 3 lambs than for those with 1 lamb. Ewes of old 

Norwegian breeds were less likely to experience CM than were ewes of 

other breeds (OR=0.59; 95% CI: 0.46-0.74). Ewes treated for CM at least 

once during the preceding three years had greater odds of CM than ewes 

without a CM history (OR=4.0; 95% CI: 3.0-5.3). 

Conclusions: The fact that the risk of CM increases considerably with 

increasing number of lambs should be taken into account in sheep 

breeding programmes. Ewes that have experienced CM are at increased 

risk of suffering a new CM episode and should be slaughtered before 

the next breeding season, even if there is no sign of chronic udder 

changes. Our effect estimates for the risk factors, which are adjusted for 

breed and unaffected by inter-flock variations, are likely to be valid 

also for other meat sheep populations. 

Key words: ewe, mastitis, risk factor, case-control study 

23 Relationship between Blue Tongue Virus Infection, Cyclicity 

and Individual Characteristics of Ewes before Autumnal 

Breeding in Texel Sheep Flocks in North of France 

C. Ponsart 1 , N. Pozzi 2 , D. Jeantils 3 , JM. Camus 4 , MC. Deloche 1 , 

S. Bareille 5 , A. Bensmaili 6 , P. Humblot 1 , B. Guerin 2 

1 

UNCEIA, R&D, Maisons-Alfort, France 

2 

LNCR, R&D, Maisons-Alfort, France 

3 

CEIA des Ardennes, Villers Semeuse, France 

4 

Chambre d'Agriculture 08, Service Technique, Villers Semeuse, 

France 

5 

Intervet-Schering-Plough, Beaucouzé, France 

6 

GDSB, Villers Semeuse, France 

In 2007, the Bluetongue (BT) outbreak from North East to the centre of 

France may have affected breeding performances of ewes in these 

regions. This study aimed to describe the effects of BT virus infection 

in 149 Texel ewes on cyclicity and individual characteristics during 

breeding. Seven flocks from the Ardennes region having at least 10 

ewes with clinical signs (CS) observed within 30 days before breeding 

and for which the breeding season started in October 2007 were 

included in the study. In each flock, two equal groups of 10-15 ewes 

(total with CS, n=77 or without CS, n=72) matched on age were bred 

with the same rams. Body condition (BC) scoring and blood sampling 

were performed in average 14±4 and 27±4 days after the start of the 

breeding period. Cyclicity was determined by progesterone assays: 

ewes for which the two progesterone measurements were

sampling. The BT status was found negative in 19 ewes (NEG, 13%, 

ELISA-/PCR-), whereas 54 were newly infected (NI, 36%, ELISA- 

/PCR+), 43 were at an acute stage (A, 29 %, ELISA+/PCR+) and 33 

immune (I, 22 %, ELISA+/PCR- ; Table 1). All ewes were cyclic, with 

no difference found in mean progesterone levels in ewes with or 

without CS (11.3±0.5 vs 11.6±0.5 ng/ml, p>0.05) and when comparing 

negative and infected ewes (NEG : 11.1±0.5 vs NI+A+I : 11.4±0.5 

ng/ml, p>0.05). The presence of CS within 30 d before breeding was 

associated with a lower BC score at first sampling (CS : 3.0±0.1 vs No 

CS : 3.5±0.1, p

aseline values. The mean HCO 3, PCO 2 and ctCO 2 increased during 

anaesthesia but decreased after AT injection. There were no significant 

alterations for haematological parameters. It is concluded that B+D+K 

anaesthesia effected the cardiorespiratoric parameters in normal 

ranges. It might be used for induction of anaesthesia. This anaesthesia 

is successfully reversed by administration of AT. 

Key words: sheep, anaesthesia, butorphanol, detomidine, Atipamezole 

55 Haematological and Clinical Biochemical Parameters for Adult 

Sheep and Goat 

A. Tschuor, B. Riond, U. Braun, H. Lutz 

University of Zurich, Department for Farm Animals, Zurich, 


Objectives: Sheep and goat husbandry has always played an important 

role in Swiss agriculture, but in recent years these animals are 

increasingly appreciated as hobby pets. This opens new diagnostic and 

therapeutic perspectives for veterinary surgeons, notably in the 

determination of haematological and clinical biochemical parameters. 

For the methods used nowadays there are no reference range values 

available for adult sheep and goats kept under central European 

conditions. Especially clinical biochemical parameters had to be 

compared with incomplete reference values, which were determined in 

the past with aged methods. Due to this, every interpretation of 

laboratory findings left incertitude. 

Material and methods: Therefore, the present study was designed to 

determine haematological (haematocrit, haemoglobin, total erythrocyte 

count, MCH, MCHC, MCV, white cell count and differentiation, 

fibrinogen, plasmatic proteins) and clinical biochemical parameters 

(bilirubin, gluc, urea, crea, albumin, cholesterol, triglycerides, AP, 

amylase, GLDH, ASAT, ALAT, GGT, CK, lipase, SDH, LDH, 

sodium, potassium, chloride, calcium, magnesium, phosphorus, iron, 

fructosamine, Alpha-1-Globuline, Alpha-2-Globuline, Beta-Globulins, 

Gamma-Globulins, A/G-ratio) in 102 goats and 102 sheep from Swiss 

flocks. The haematological examination was done using a Cell Dyn 

3500 ® (Abbott AG, Diagnostics Division) and the examination of the 

biochemical parameters were done by a Cobas Integra (Roche 

Diagnostics). 

Results: Significant differences were found between the two species 

especially regarding erythrocyte count, mean erythrocyte volume, 

haemoglobin content as well as leukocyte count. There were also 

significant differences between this new obtained reference values 

compared to the aged ones from central Europe. 

Conclusion: It is therefore of utmost importance to discriminate 

between sheep and goats when interpreting laboratory findings by 

means of actual reference values. Additionally, the study provides a 

literature based overview explaining the significant differences 

between these two species in comparison to the bovine reference 

values. 

56 Impact of Bluetongue Virus Serotype 8 on Rams' Semen 

Quality 

N. Kirschvink 1 , M. Raes 1 , C. Saegerman 2 

1 



2 

University of Liege, Department for Infectious Diseases - 

Epidemiology; Faculty of Veterinary Medicine, Liege, Belgium 

Bluetongue virus (BTV) is reported to induce transient infertility in 

infected ruminants. The aim of this study was to assess the effects of 

BTV serotype 8 on semen quality of rams tested at several occasions 

after infection and by comparing them to healthy control rams. 11 

healthy rams tested prior to the BTV outbreak in Belgium were used as 

controls (C) and 12 rams with natural infection by BTV serotype 8 

(BT) were included in this study. BT rams were tested on 2-6 occasions 

(BT-S1 to BT-S6) 5 to 138 days after development of clinical signs of 

BTV. BTV infection was serologically confirmed. Semen was 

evaluated for motility (score ranging from 0-5), spermatozoid 

concentration and viability (% of normal living [NL]; normal death 

[ND]; abnormal death [AD] spermatozoa). 

Results are shown as mean ± SD. 

These results demonstrate the important impact of serotype 8 BTV on 

rams’ semen quality. 85 days after development of clinical signs of 

BTV, the variables tested did no more significantly differ from control 

animals, suggesting that full recovery of semen quality occurred. 


Controls BT-S1 BT-S2 BT-S3 BT-S4 BT-S5 BT-S6 

Motility 4.5±0.2 0.8±0.5* 1.3±1.9* 2.4±1.9* 3.3±1.6* 4.0±0.9 4.5±0.3 

Conc (10 9 

sperm/ml) 3.2±0.7 1±0.7* 1.2±1.4* 1.2±0.9* 1.5±0.8* 2.2±0.9* 2.6±0.6 

% NL 75±6 9±10* 27±32* 46±32* 54±25* 63±16* 76±8 

% ND 21±7 48±12* 37±22* 33±21* 30±10* 29±11* 20±7 

% AD 4±3 43±11* 37±28* 21±25* 16±23* 9±13 4±4 

* significantly different from respective control-value, p

group 2) received blood 3) received I/V hypertonic saline 4) received oral 

isotonic electrolyte solution and 5) received intravenous hypertonic saline 

followed immediately by oral electrolyte solution. Hemorrhagic shock 

was induced by constant removing of blood over 15-20 minutes. The 

amount of the withdrawn blood ranged between 532 ml to 616 ml. 

depending upon body weight of each goat. Immediately before 

hemorrhagic shock induction, the baseline line values were recorded 

including heart rate, ECG, respiratory rate, rectal temperature, body 

weight and a maximum clinical score (MCS), Hematocrit, blood gas 

analysis) pH, Pco 2, Po 2 ), serum biochemical analysis (Na, K, Cl, Ca, 

Phosphate, total protein, albumin and creatinine concentration), blood 

lactate and plasma volume changes. These parameters were also 

determined at the time of entry into the treatment phase, 30 minutes, 2 

hours, 8 hours, and 24 hours after treatment. 

Results and Conclusions: results indicated that intravenous 

administration of hypertonic saline solution together with oral 

administration of electrolyte solution in hypovolemic goats caused 

immediate and sustained effect. It was superior to administration of 

either the two solutions alone and induced the same effect as whole 

blood. Meanwhile resuscitation with Hypertonic saline solution alone 

resulted in only transient resuscitation. 

Key words: goat surgery, hypovolemic, shock 

93 Estimation of Gestational Age in Egyptian Goats by 

Ultrasonographic Fetometry 

A. Karen 1 , E. Fatouh 1 , S. Shokr 2 

1 

Faculty of Veterinary Medicine, Theriogenology, Kafr El-Sheikh, 

Egypt 

2 

Faculty of Veterinary Medicine, Anatomy and Embryology,Ismailia, 

Egypt 

The present study aimed to determine the earliest day at which the signs 

of pregnancy can be detected by transrectal (TR) and transabdominal 

(TA) scanning and to estimate the gestational age in Egyptian goats by 

ultrasound measurement of embryonic or fetal parts throughout 

pregnancy.Fifteen pregnant Egyptian goats were used in the present 

study. The date of their mating was considered as Day 0 for calculating 

the gestational age. The does were transrectally scanned by using a 7.0 

MHz vector probe (Acuson 128 XP/10 computed scanner) at Day 10 

post mating on alternate days until Day 25 and then once at 3-5 day 

intervals until day 50. Linear and convex (3.5 to 5 MHz) probes were 

used for TA scanning which were carried out from Day 25 to Day 130 

at 3 to 5-day intervals. The days after mating at which the embryonic 

vesicle and the embryo proper with a beating heart firstly imaged by TR 

and TA scanning were recorded. After imaging the embryo or the fetus, 

the following parameters were measured: length of the embryo or fetus 

(CRL), heart rate (FHR), biparital diameter (BPD), trunk diameter 

(TD), placentome size (PS), umbilical cord diameter (UCD) and femur 

length (FL).The average of days at which the embryonic vesicle was 

firstly determined by TR and TA scanning was 16.98±1.97 and 

27.87±3.48, respectively. The embryo proper with a beating heart was 

firstly determined by TR and TA scanning at an average of 22.36±2.66 

and 30.36±4.75 days, respectively. All the fetal measures were 

significantly (P

immunization with either the currently produced vaccine (A&M vaccine) 

or with a wild-type contagious ecthyma viral strain does not result in 

cross-protection. Antigenically dissimilar CE viral strains may have 

contributed to vaccine failures, thus contagious ecthyma vaccines should 

be multivalent or include locally occurring strains. 

Key words: contagious ecthyma, goats, vaccine, immunology 

96 Prevalence and Determinants of Ixodids in Domestic Goats 

Lower Punjab (Pakistan) 

S. Sajid 1 , Z. Iqbal 1 , N. Khan 1 , G. Muhammad 2 

1 

University of Agriculture, Faisalabad, Parasitology, Faisalabad, 

Pakistan 

2 

University of Agriculture, Faisalabad, Clinical Medicine and 

Surgery, Faisalabad, Pakistan 

Tick infestation is a serious nuisance in small ruminants of developing 

countries like Pakistan. The objective of the current study is to 

determine the prevalence and various determinants of tick infestation in 

caprines of lower Punjab (Pakistan). For this purpose, a total of 700 

goats of various breeds, age and sex were randomly selected from each 

of district Muzaffargarh and Layyah of lower Punjab. The selected 

herds were visited thrice a month in order to collect all the relevant 

information including the species, breed, age and sex of host, infested 

number of animals, type of housing and average number of ticks per 

animal on a pre-designed questionnaire. Ticks were collected and 

processed through standard procedure for their taxonomic 

identification. Hyalomma anatolicum and Rhipicephalus sanguineus 

species were identified from the subjected hosts. The overall 

prevalence of ticks was 51.6% (723/1400) in both the districts. The 

highest month-wise prevalence in Districts Layyah (53.42%; 374/700) 

and Muzaffargarh (60.8%; 426/700) were found in July and lowest in 

November (3.14%; 22/700) and October (3.14%; 22/700), respectively. 

No significant (P = 0.760) differences in the prevalence of tick 

infestation as calculated through Z-test (Z = 0.31) were found between 

district Layyah and Muzaffargarh. However, significant differences 

were found in prevalence percentages of various tehsils of Layyah and 

Muzaffargarh. Regarding determinants, prevalence of tick infestation 

was found higher in males, older animals and Beetal breed goats. The 

preferred sites of ticks were inside and external ear. The results 

provided a better understanding of the status of tick infestation in the 

domestic goats of the study area and will be helpful in the planning of 

control programme against ticks. 

Key words: prevalence, ticks, goats, ixodids, Pakistan 

97 Effect of Boron on Haemogram and Biochemical Profile of 

Urine in Buffalo (Bubalus Bubalis) Calves Fed High Fluoride 

Ration 

V. Bharti 1,2 , M. Gupta 1 

1 CCS Haryana Agricultural University, Hisar, Department of 

Veterinary Physiology, Hisar, India 

2 Indian Veterinary Research Institute, Division of Physiology & 

Climatology, Bareilly, India 

The present study was carried out with the objective of ameliorative 

effect of boron (B) on high fluoride (F) intake. As per Randomized 

Block Design, twelve male Murrah buffalo calves of 6-8 months age 

were divided into three groups of four calves in each group. They were 

fed conventional concentrate mixture, roughage and supplemented 

with sodium fluoride (NaF, 60ppm elemental F) alone or in 

combination with borax (Na 2B 4O 7.10H 2O, 140ppm elemental B) for 

90 days. Blood sample were taken on 0, 30 th , 60 th and 90 th day for 

analysis of Hb, TEC and PCV. Urine were collected at the end of 

experiment (90 th day) and analyzed for alkaline phosphatase activity 

(ALP) and minerals (viz, Ca, P, Fe, Zn and Cu).This level of fluoride in 

ration caused a significant (p

esulted efficacious after 3h of incubation in respect to control 

condition. Moreover 10 -6 M Nx at 1 and 3h statistically increased 

(p

decrease in animal aggregation in pastures 60 days age may be 

suggested for weaning weight of Farahani male lambs for fattening. 

Key words: weaning age, feedlot traits, Farahani male lambs 

740 Effectiveness of Oestrus Induction Treatments in Sarda 

Breed Lambs during Non-breeding Season 

M. Ferrari 1,2 , G. Catone 1 , M. Baroni 1 , P. Petralia 1 , S. Chiodi 1 , 

B. Castellucci 1 , L. Ferrari 2 , A. Malfatti 1 

1 

University of Camerino, Italy, Department of Veterinary Medicine, 

Matelica (MC), Italy 

2 

Federal University of Paraná (UFPR), Department of Veterinary 

Medicine, Curitiba, Brazil 

Despite the interest and opportunity to use yearling lambs for 

ameliorating reproductive performance of flock and as substitutes for 

unproductive adult ewes, there is a lack of information about onset of 

inducted oestrus in Sarda breed lambs during non-breeding season. The 

purpose of this work was to evaluate the efficacy of four different 

procedures for oestrus induction out of reproductive season in Sarda 

breed lambs. The experiment was performed during early spring season 

(end of March) on a farm located in a hill area (400 m above sea level) 

in The Marche region (Central Italy). Ninety and two 11 month-old 

Sarda lambs were used. The animals were randomly chosen and 

divided in four groups designed GI (n=22), GII (n=22), GIII (n=24) and 

GIV (n=24). Each group remained in separated pens during the whole 

experiment. The animals were fed with good quality hay and 300 grams 

of fodder by head in a daily basis and have no contact (visual, sound, 

smell) with males during at least 2 months before the start of the 

experiment. All lambs were treated with intravaginal sponges 

impregnated with 40 mg of FGA (Cronogest ® ) which were removed 

after 9, 12, 14 and 14 days after insertion, respectively for GI, GII, GIII 

and GIV. PMSG was injected (i.m.) into all lambs at the moment of 

sponge withdrawal (500 UI for GI, GIII and GIV and 300 UI for GII). 

GI and GIII lambs were injected (i.m.) with 125 µg of cloprostenol at 

the moment of sponge introduction. Rams with the chest painted by a 

dye mixed in an oil base were introduced after 24 hours from sponge 

withdrawal in all groups and the onset of oestrus was verified by 

inspection of mating rump marks present on lambs every 12 hours from 

ram introduction to 72 hours after. Respectively for GI, GII, GIII and 

GIV, the onset of oestrus was 81,8% (n=18), 72,7% (n=16), 87,5% 

(n=21) and 62,5% (n=15) and the pregnancy rates were 50%, 46%, 

58% and 46%. From these results, we could conclude that both oestrus 

and pregnancy rates were not affected by the duration of the sponge 

treatment nor by the PMSG dose and that the cloprostenol 

administration resulted in a higher number of mated lambs, considering 

all the groups (85% vs 67%; p= 0.05) or only the 14 days groups (GIII 

vs GIV; p< 0.05). 

Key words: oestrus, induction, lambs, Sarda breed 

741 A Case Report of Sporadic 10 Ovine Menigoencephalitis in a 

Herd in Sao Paulo State, Brazil 

L. Gregory 1 , E. Meira Jr. 1 , A. Nassar 2 , E. Scarcelli 2 , E. Villalobos 2 , 

M. Lara 2 , A. DellaLibera 1 , A. Safatle 1 , P. Barros 1 , C. Del Fava 2 


2 Instituto Biológico, Centro de Sanidade Animal, Sao Paulo, Brazil 

In a flock of 250 Santa Ines ovines proceding from a farm in Bragança 

Paulista, Sao Paulo state, Brazil, it was investigated the presence of a 

neurological acute disease in ten adult animals between October and 

November 2007. The clinical symptomatology included head 

deviation, opisthotonus, nystagmus, ataxia with consistent falling to 

one side, `paddling` involuntary movement and stiff limbs. The 

animals also presented hyperthermia, blindness, letargy and muscle 

stiffness with restrained movements. The sheep with visual impairment 

was submitted to ophthalmic exam. The eyes presented calmness and at 

the direct and indirect ophthalmoscopy, were not possible to detect any 

retinal impairment. The animal was submitted a full field 

electroretinogram. The procedure was performed according to the 

standard protocol recommended by the ISCEV (International Society 

for Clinical Electrophysiology of Vision), which is constituted of five 

responses. The ERG showed reduced amplitude in all five responses, 

when compared with healthy sheep. With these results the visual 

impairment was not explained by retinal diseases. The main clinical 

suspicion was Listeria monocytogenes because, at the farm, all the 

animals were being fed with silage from a newly open silo. From ten 


animals examinated, four died and other two were euthanised at the late 

stage of the disease. In the remaining animals it was carried out general 

and specific clinical exams. The complementary exams were 

considered, therefore the disease must be differentiated from Rabies 

(IFD), Listeria monocytogenes (PCR and culture) and Toxoplasmosis 

(serology). The serological and bacteriological exams were negatives. 

At the liquor exam was observed large presence of monocytes, 

indicating an almost complete monocitosis. The histological lesions 

were predominantly unilateral with typically inflammatory response as 

described in meningo-encephalitis cases. It was not possible find the 

etiological agents in this case demanding more studies in sheep 

encephalitis causes in Brazil. 

Key words: sheep, encephalitis, clinical of ruminants 

742 Retrospective Study: does Vaccination against Footrot 

Improve Reproduction Performances in Ewes? 

C. Marcerou, B. Bolkaerts, C. Baricalla, N. Kirschvink 



The aim of this retrospective study was to assess whether vaccination 

against footrot (Fusobacterium necrophorum and Dichelobacter 

nodosus) by use of a vaccine with 10 valences against Dichelobacter 

nodosus would affect the reproduction performances in a flock of adult 

ewes with a high prevalence of footrot. 

Data obtained from a sheep flock of adult Texel, Ile de France and 

Texel-Ile de France ewes prior to vaccination (Pre: 2005-2006) were 

compared to data obtained after vaccination (Post: 2006-2007). Except 

instauration of footrot vaccination, the flock management remained 

identical: antiparasitic control, reproduction periods (autumn), food 

availability and mean parity (3.6±1.5) did not differ between both 

periods. The following variables were assessed: number of oestrus 

cycles [OC] necessary for gestation (calculated individually by 

considering the moment of introduction of the rams into the flock and 

the moment of lambing), fertility, fecundity, prolificity and % of 

abortions (defined as non lambing ewes after confirmed gestation by 

ultrasound diagnosis). 

Results: Data are shown as mean ± SD (if appropriated). 

OC Fertility Fecundity Prolificity % 

(%) (%) (%) abortions 

Pre 2.64±2.55 78 126±86 1.61±0.61 17 

(n=231) 

Post 2.18±2.06* 96* 150±61* 1.56±0.54 3* 

(n=204) 

* Significantly different from respective Pre-value, p

Material and methods: This study was conducted on 500 female and 

500 male, 1-5 year-old ages Ghezed sheep in four seasons. To assay of 

vitamin A and Beta carotene 5ml blood was taken by using venoject 

tube from jugular vein of each sheep, and then separated its serum by 

centrifuge. Liver samples were provided after slauthering them. The 

levels of vitamin A and Beta carotene of blood and liver were measured 

by spectrophotometer. 

Results: The mean of blood and liver vitamin A and Beta-carotene 

were higher in summer than winter (P0.05) table 1. Blood and 

liver vitamin A were more at 2 year old ages sheep but blood and liver 

carotene levels were high at 4 and 5 year-old ages respectively (P0.05) but the 

levels of Beta-carotene were more at female sheep (P< 0.05). 

Table 1. Mean of blood and liver vitamin A & Beta-carotene in 

different seasons 

measured Blood Liver Blood Liver 

items vitamin vitamin Beta Beta 

season A µg/dl A µg/g carotene µg/dl carotene µg/g 

Spring 32.98 36.62 17.07 20.90 

Summer 38.23 40.03 20.36 24.61 

Autumn 35.20 37.12 18.01 22.34 

Winter 30.57 31.54 15.89 19.20 

S= significant effect at (P0.05) from those found with the MTDPL or UTDPL. 

The DOMI (16g/kgW 0.75 ) of the MTDPL was significantly (P0.05) from those found with the UTDPL or the MTDPL. The DM 

and OM digestibility of the MTDPL (37.2%) were higher (P0.05). The results indicated that 

the voluntary feed intake and organic matter digestibility of date palm 

dry leaf improved as a result of mycelial treatment but this advantage 

diminished after harvesting the mushroom. 

Key words: Pleurotus florida, nutritive value, digestibility, date palm 

dry leaf 

745 Fattening Performance, Carcass Traits and some Meat 

Quality at Different Slaughter Weights of Bafra Lambs 

A. Yakan 1 , N. Ünal 2 

1 University of Ankara, Institute of Health Science, Department of 

Animal Science, Ankara, Turkey 

2 University of Ankara, Faculity of Veterinary Medecine, Department 

of Animal Science, Ankara, Turkey 

The purposes of this research were to investigate the fattening 

performance, carcass traits and some meat quality at different 

slaughter weight of Bafra lambs (Chiosx Karayaka B1). Data were 

collected from 40 head Bafra lambs, approximately live weight 20 kg 

in Amasya- Gökhöyük State Farm (in Turkey) from May to December 

2007. Feeding during fattening period was consisted of a concentrate 

mixture available ad libitum and 300 g of alfalfa hay per lamb per day. 

When lambs reach at 30, 35, 40 and 45 kg, 6 lambs of each slaughter 

weight were slaughtered for determining of slaughter, carcass traits 

and meat quality. The average daily weight gain and feed conversion 

efficiency were 283, 251, 241 and 251 g; 3.460, 3.653, 4.966 and 

5.292 kg dry matter intake/ kg body weight gain for each slaughter 

weight group, respectively. In each slaughter group, hot carcass 

percentages, dressing percentage were 51.92, 53.48, 54.96 and 54.76 

% (P0.05); loin eye 

areas were 13.616, 13.841, 15.091 and 16.508 cm 2 ; ratios of lean on 

carcass were 50.22, 48.60, 50.53 and 45.76 % (P

747 Effect of Dietary Supplementation of Tinospora Cordifolia on 

Metabolic Profile, Antioxidant Status and Immune Response 

in Lambs under Semi-Intensive System 

D. Nagalakshmi 1 , K. Dhanalakshmi 2 

1 College of Veterinary Science, Department of Animal Nutrition, 

Hyderabad, India 

2 College of Veterinary Science, Department of Microbiology, 

Hyderabad, India 

Twenty four, male Nellore lambs (9-10mon) of uniform body weight 

(BW) reared under semi-intensive system were randomly allotted to 4 

groups (6 in each group) in completely randomized design to evaluate the 

anti oxidant and immunomodulatory effect of Tinospora cordifolia. The 

reference lambs received the basal concentrate mixture (no herb) and the 

experimental lambs received 60, 90 and 120mg/kg BW of Tinospora 

cordifolia, respectively for a period of 90days. The blood haematological, 

biochemical and oxidative enzyme activity measured at 0, 45 and 90d 

indicated higher (P

of offspring equalled to the threshold phenotype of dam. For each 

heritabilities, 10 replicates were run. Having generated all the data, 

genetic and environmental parameters, such as means and variances of 

genetic, underlying phenotypic and threshold phenotypic were 

calculated for each generation. The underlying phenotypic data were 

analysed by ASREML version 2.00a with animal model. The threshold 

phenotypic data were analysed by MATVEC using threshold model. 

Results: Having calculated genetic parameters for all generations, it was 

found that genetic and phenotypic means increased as well as threshold 

mean. The results showed that response to auto selection depended on 

dam distribution for the threshold phenotype. Furthermore, heritability 

remained constant across generations and there was no significant 

difference in compare with the base generation. Analysing threshold data 

resulted that corresponding heritabilities at theoretical 0.10, 0.25 and 0.50 

for underlying phenotype in base population were estimated to be 0.0643, 

0.1697 and 0.2807, respectively. 

Conclusions: Auto selection increases the genetic, phenotypic and 

threshold means, but had no effect on heritability. However, analysis of 

threshold data with threshold model resulted in under-estimate 

heritabilities compare with their corresponding underlying heritabilities. 

Key words: simulation, auto selection, population parameters, sheep 

752 Evaluation of Somatic Cell Counts (SCC) for the Diagnosis of 

Mammary Infection in Lacaune Sheep 

V. Gomes 1 , M. Blagitz 2 , K. Madureira 1 , A. Gomes 3 , A. Della Libera 2 

1 Faculdades Anhanguera, Medicina Veterinária, Leme, Brazil 


3 Universidade Estadual do Norte do Parana, Medicina Veterinária, 

Bandeirantes, Brazil 

Somatic cell count (CCS) is an important tool in the diagnosis of 

mammary infection and inflammatory process in ruminants. However, 

little is known about the use of cell count techniques and its usefulness in 

sheep, mainly in relation to Lacaune dairy sheep. The present study 

analyzed 200 samples of milk collected from the halves of 100 Lacaune 

sheep in different phases of lactation. The first aliquot was used in 

automatic cell counts by means flow citometry; the second aliquot was 

used in direct microscopic counts using methyl green-pyronin Y and 

Rosenfeld; the third aliquot was used in bacteriological examination. 

From the 200 samples analyzed, 14 were positive at the bacteriological 

examination, 2 samples were contaminated and 184 were negative. 

Bacteria isolated and identified were: Staphylococcus hyicus (n=1); 

Staphylococcus Kloosii (n=7); Staphylococcus haemolyticus (n=5); and 

Staphylococcus epidermidis (n=1). Results for automatic cell counts were 

0.15 and 0.113x10 6 ; for microscopic cell counts using methyl greenpyronin 

were 0.126 and 0.320 x10 6 ; and microscopic counts using 

Rosenfeld were 0.127 and 0.851 x10 6 , for samples positive and negative 

in the bacteriological examination, respectively. Results for cell counts 

showed significant differences between the groups of positive and 

negative animals only when methyl green-pyronin Y and Rosenfeld were 

used, making it necessary to be very careful in the choice of the adequate 

technique for the detection of mammary infection. 

Key words: sheep, somatic cell counts, infection, milk 

753 Control of Eimeria spp Natural Infections in Santa Ines 

Lambs Reared under Semi-intensive Exploitation Condition 

in Brazil by Treatment with Toltrazuril 

T. Silva1 , EJ. Facury filho2 , PM. Ferreira2 , AU. Carvalho2 , 

A. Nunes3 , FH. Albuquerque4 1 ReHAGRO, Ovinocultura, Belo Horizonte, Brazil 

2 Escola de Veterinária da UFMG, Ovinocultura, Belo Horizonte, 

Brazil 

3 Prefeitura Municipal de Belo horizonte, Vigilância Sanitária, Belo 


4 Embrapa Caprinos, Pesquisa E Extensio, Sobral, Brazil 

The metafilactic use of toltrazuril (Baycox) in preventing Eimeria spp 

infection in naturally infected lambs was evaluated.Forty three lambs 

were divided into 4 groups treated with 20mg of toltrazuril: group 1: 

treated at the 4 th week of age; group 2: treated at the 4 th and 8 th weeks 

of age; group 3: control; group 4: treated at the 2 nd week of age. Fecal 

samples were collected weekly until lambs were 12 weeks old, a last 

sample was collected at the 16th week. Oocysts were quantified in each 

sample according to Gordon and Whitlock’s technique modified by 

Ueno and Gonçalves (1998). Eimeria species were identified according 

to morphological characteristics of oocysts and sporocysts (Levine, 

1985; Reginsson and Richter, 1997). The oocysts excretion curves 

were similar for all treatments, but not the time in which each event 

happened. Control group lambs had a gradual rise in oocysts excretion 

after the 3 rd week of age. Two peeks occurred until the 12 th week of 

age, one at 7 th (125985 OOPG) and other at 10 th week (101657 OOPG). 

For the other groups this rise began between the 3 rd and 6 th weeks. 

Groups 2 and 4 also had 2 peaks, respectively, at the 8 th (49708 OOPG) 

and 12 th (47933 OOPG), and at 9 th (220521 OOPG) and 12 th (108416 

OOPG) weeks of age. Group 1 had only 1 peak at the 10 th week 

(119654 OOPG). After the 10 th week of age oocysts output of all 

groups gradual and continuously reduced, maintaining low levels until 

the 16 th week of age. Control group lambs had the highest oocysts 

output when compared to other groups (P

755 Study of Pulmonary Adenomatosis in Sheep in West 

Azarbyjan 

Gh. Moghaddam1 , J. Ashrafi helan2 , A. Nematollahi2 1 Faculty of Agriculture,University of Tabriz, Animal Science, Tabriz, 

Iran 

2 Faculty of Veterinary Medicine, University of Tabriz, Department of 

Pathobiology, Tabriz, Iran 

Objective: sheep pulmonary adenomatosis is a contagious disease 

produced by a tumor in the lung. Its morbidity and mortality rate are 

2% and 100% respectively. The purpose of present study was 

estimation of affected sheep by lung carcinoma, because these 

knowledge need to control it. 

Material and methods: This study was conducted on 50 apparent 

health and 50 affected sheep pulmonary. Lungs were provided from 

Tabriz slaughter house. All lungs were inspected macroscopic and 

histopathology. 

Result: 23 lungs associated with tumor were observed at present study. 

20 cases were related to affected group and 3 cases to apparent health 

group. Pulmonary carcinoma was present in both lungs. Any 

adenomatosis lesions were not noticed on cardiac and intermediate 

lobes, but diaphragmatic lobes had most lesions (tab.1). Affected lungs 

were considerably enlarged and heavier than normal. Carcinoma 

lesions were seen large, firm and gray masses or hard white nodules. 

The trachea and bronchi often contained clear and foamy fluid. Bronchi 

had riddle appearance in cut surface due to hyperplasic process. The 

characteristic changes noticed was hyperplasia and hypertrophy of 

alveolar epithelium and accumulation of large mononuclear cells in the 

alveolar spaces. 

Table 1. presence of tumors in lobes of lungs 

Item All Apical Cardiac Intermediate Diaphragmatic 

lobes lobe lobe lobe lobe 

Right lung 2 3 0 0 16 

Left lung 1 3 0 0 19 

Discussion: Presence of high adenomatosis lesions in sheep lungs 

indicates that this disease is spreading in Iran especially in west azar 

byjan around. According to economic losses and public health 

important of sheep pulmonary adenomatosis, programming for its 

control and prevention is suggested. 

Key words: sheep, adenomatosis, carcinoma, lung 

756 Impact of Bluetongue Virus Serotype 8 on Anti-VP7 Antibody 

Levels in Serum and Colostrum of Ewes and in Suckling 

Lambs 

A. Lamerant, B. Bolkaerts, C. Baricalla, N. Kirschvink 



The aims of this study were to perform a semi-quantitative assessment 

of anti-VP7 antibodies (Ab) (1) in serum (T0-ewe) and colostrum of 

lambing ewes that were between 2-8 weeks of gestation when they 

were naturally exposed to bluetongue virus (BT) serotype 8 in summer 

2007; and (2) in serum of lambs at birth (T0-lamb) and 24h after 

colostrum intake (T24-lamb). Among the 58 pregnant ewes, 9 had 

shown clinical signs of BT between 19-47 days of gestation. At 

lambing, blood and colostrum was sampled in ewes and in their lambs 

(n=114) prior to colostrum intake and 24 h after colostrum intake. Anti- 

VP7 Ab was semi-quantitatively assessed by adapting a competition 

EIA (ID Screen, Blue Tongue Competition, France). 

Results are shown as means ± SE in function of positive (BT+) and 

negative (BT-) serology (% of inhibition) and in form of arbitrary units 

of anti-VP7 Ab. 

Positive and significant correlations were found between serum and 

colostrum Ab levels in ewes (r=0.32, p

Materials and methods: ovaries were collected from a local abattoir 

and transported within 3 h to the laboratory in awarm saline solution 

(30-35 °C), supplemented with 100 µg streptomycin sulfate/ml. 

Cumulus-oocytecomplexes (COC’s) were obtained by slicing of 

follicles, washed in TCM-199 modification with NaHCo 3 and 

supplemented with 50 µg/ml gentamycin, and 0.25 mM sodium 

pyruvate without any serum supplementation. The COC’s were 

randomly divided into four groups. Group 1 (n = 105) COC’s were 

fresh control and cultured in TCM-199 medium without serum 

supplementation. Group 2 (n = 108) COC’s were washed five times and 

cultured in TCM-199 medium supplemented with 10% MS. Group 3 (n 

= 112) COC’s were washed five times and cultured in TCM-199 

medium supplemented with 15% MS. Group 4 (n = 114) COC’s. After 

38-42 h of IVM, oocytes were denuded with the aid of 0.1% 

hyaluronidase and passing them through a fine pipette, fixed for 24-48 

h in a mixture of acetic acid and alcohol (1:3) at room temperature, 

stained for 10 min with 1% (w/v) orcein in 45% acetic acid and 

examined for the evidence of different stages of maturation. 

Results: In this study significantly higher (p < 0.05) maturation rates of 

oocytes (69-72%) were observed in all concentrations of Mare serum 

compared to those without serum supplementation. However, no 

significant difference was observed between the 10, 15% and 20% 

serum supplemented group. 

Conclusion: In conclusion, it can be said that the supplementation of 

10% MS in TCM-199 medium could be used to achieve optimal 

maturation rate. 

Key words: sheep, mare serum, follicle slicing, oocyte maturation, 

ovary 

760 Evaluation of Decoquinate Influence on Fast Growing Young 

Awassi Lambs: Field Trial 

H. Leibovich, A. Ben Zeev, A. Richard 

Alpharma, Animal Health Division, Verrieres, France 

Most of the sheep kept under intensive conditions suffer some level of 

coccidiosis. Most young sheep and goats undergo coccidial infestation, 

which is known to cause diarrhea, and can also silently decrease growth 

and so impair performance significantly. Decoquinate is a non-antibiotic 

synthetic molecule, active against certain protozoa: coccidia, 

cryptosporidia, toxoplasma, neospora. The purpose of this trial was to 

evaluate the ability of 1 mg/kg BW per day décoquinate to control 

Coccidia in young lambs and its influence on lamb’s growth performances 

under intensive management in Israel. Experiment was conducted on a 

commercial Awassi dairy farm (1500 ewes) located in Kibbutz Ein- 

Harod. Experimental lambs were born to synchronized ewes lambed on 

May 2005 - an out of season time. 92 Male and Female lambs, were 

separated from their dams just after birth, fed colostrums for 4 days, and 

then were assigned to the treatment or control groups, in order to design 

similar groups according to birth weight, litter size and sex. Lambs were 

offered ad lib of reconstituted milk until the age of 31 days. Deccox 

included in lamb’s diet from the age of day 4 until 91 days. Body weights 

were collected at birth, 31 days (weaning), 60 days, 91 days, 132 days, and 

168 days. Milk replacer and concentrates were offered fresh daily for ad 

lib., residues were collected and recorded daily. Oocyst counting on 

faeces samples from 10 lambs from each group collected on days 14, 28, 

and 91. Lambs average weight was 4.7 kg, consumed during suckling 

period 6.9 kg per lamb in the Deccox group and 9.5 kg in the control 

group. Weaning weights were 11.5 kg and 10.4 kg for control and Deccox 

groups. After weaning, the Deccox lambs increased daily food intake to 

an average of 38.2 kg compare to 29.7 kg for the control group for the age 

31 - 91 days. The Deccox lambs weight 24.2 kg compare to 21.5 kg of the 

control group. During the consecutive period, the Least Square means of 

the Deccox lamb’s weight on days 63, 91, 133, and 168 were 16.6, 24.2, 

35.1, 49.2 kg compare to 15.8, 21.5, 32.0, and 43.9 kg for e control group. 

Food conversion rate was 2.7 for the Deccox group, and 3.0 kg for the 

control. Numbers of oocyst counted on day were 2650 for the control and 

460 in the Deccox. These results confirm that the decoquinate treatment 

improves performance of the fast growing lambs - increased growth rate 

and food conversion rate of the lambs. 

761 Pestiviral Infections in Sheep in Sao Paulo State, Brazil 

E. Pituco 1 , C. Ribeiro 1 , E. De Stefano 1 , N. Galleti 1 , M. Oliveira 1 , 

A. Nogueira 2 

1 Instituto Biológico, Centro de Pesquisa e Desenvolvimento de 

Sanidade Animal, Sao Paulo, Brazil 

2 Apta Regional, Pólo Regional do Extremo Oeste / UPD de 

Aracatuba, Aracatuba, Brazil 

To determine the prevalence of Pestivirus infections in sheep, 

antibodies to Bovine Viral Diarrhoea Virus (BVDV) using the NADL 

strain of BVDV (BVDV-NADL), were investigated. The number of 

samples that had to be studied for correct statistical evaluation was 

determined with an estimated prevalence of 50%, confidence interval 

of 95% and relative precision desired of 7%. As a result, 64 flocks were 

randomly selected. Only ewes over one year old were included. Serum 

samples of 1306 ewes, from three regions of Sao Paulo State namely 

Bauru, Sorocaba and Araçatuba from healthy ewes that had not been 

vaccinated for BVDV and BDV were analyzed using virus 

neutralization method recommended by the OIE. Ewes with antibody 

titers equal or greater than 4 were considered as positive. The overall 

percentage of animals with antibodies to BVDV was 2% (27/1306), in 

nine flocks 14% (9/64) and the titers ranged between 4 and 64. There 

were highly significant geographical variations in antibody prevalence 

ranging from 0% in Bauru and Sorocaba regions to 26,5% in Araçatuba 

region. These results show that pestivirus infections are present in 

Brazilian sheep, although at a low rates. The presence of reactors 

sheeps only in Araçatuba may be attributed to the nearness of the Mato 

Grosso do Sul State, where there is movement of relatively large 

number of cattle, and also because cattle and sheep are in close 

proximity, which is in contrast to the others two regions analyzed. 

Further studies are being undertaken to detect the viremia and also to 

evaluate the cross neutralization on all positive samples with BDV, and 

to identify the causative agent. This will be necessary because 

Pestiviruses are not strictly host specific, and BVDV infections have 

been found to occur frequently in sheep. 

Key words: Pestivirus, serology, antibodies, prevalence, sheep 

762 Determination of Myocardial Degeneration due to White 

Muscle Disease in Lambs by Qualitative Cardiac Troponin 

Kits 

V. Gunes1 , K. Ozcan2 , M. Citil3 , A. Onmaz1 , H. Erdogan3 1 University of Erciyes, Faculty of Veterinary, Internal Medicine, 

Kayseri, Turkey 

2 University of Kafkas, Faculty of Veterinary, Pathology, Kars, 

Turkey 

3 University of Kafkas, Faculty of Veterinary, Internal Medicine, 

Kars, Turkey 

White muscle disease (WMD) is one of the common problems of 

lambs. It develops due to Vit-E and Selenium deficiency. Hyalin 

degeneration and severe necrosis occurs in myocardial. This leads to 

acute heart failure. The aim of this study was to determine the value of 

human cTn-I and cTn-T immunochromatographic strip kits in the 

diagnosis of myocardial degeneration due to WMD in lambs. For this 

purpose, 10 lambs with acute WMD, mean age 3.5±1 weeks old (range 

2-8 weeks), and, 6 clinically healthy lambs, mean age 4±1 weeks old 

were used. Routine clinical examinations, troponin analyses and 

necropsy were performed in the lambs with acute WMD. Only blood 

samples were collected from healthy lambs for cTn-I and cTn-T assays. 

Cardiac Tn-T was determined by Tropt Sensitive Rapid Assay (Roche- 

Germany). Cardiac Tn-I tests was carried out using Card-I Kit Combo 

Test (Aboatech-Finland) in serum of the animals. Cardiac Tn-I and 

cTn-T tests were determined as positive in the all lambs with WMD, 

but these tests were negative in blood samples collected from healthy 

lambs. In necropsy findings, the cardiac and skeletal muscles were 

observed to be affected in varying degrees by chalky white lesions. 

Histopathological examination revealed necrosis and calcification in 

the myocardium and skeletal muscles in lambs with WMD. 

Histopathological findings of heart muscle and positive cTn results 

were correlated in cases with WMD. In conclusion, Troponin assays 

(cTn-T and cTn-I) may be sensitive and specific marker for small 

ruminants with myocardial injury, and can be used to predict the future 

cardiac disorders in ambs. 

Key words: lamb, white muscle disease, cardiac troponins 

763 The Effect of Oxfendazole + Oxyclozanide Paste and Tablet 

Formulations on Parasite Burden and Metabolic Status of 

Sheep 

V. Günes 1 , A. Inci 2 , F. Uyanik 3 , A. Yildirim 2 , N. Altug 4 , M. Eren 3 , 

A. Onmaz 1 , C. Gelfert 5 

Sheep, Goat and Buffaloes 123

1 University of Erciyes, Faculty of Veterinary Medicine, Department 

of Internal Medicine, Kayseri, Turkey 

2 University of Erciyes, Faculty of Veterinary Medicine, Department 

of Parasitology, Kayseri, Turkey 

3 University of Erciyes, Faculty of Veterinary, Department of 

Biochemistry, Kayseri, Turkey 

4 University of Yuzuncu Yil, Faculty of Veterinary Medicine, 

Department of Internal Medicine, Van, Turkey 

5 University of Veterianary Medicine, Clinic for ruminants, Vienna, 

Austria 

The aim of the present study was to evaluate the effects of anthelmintic 

treatment with Oxyclozanid+Oxfendazole in different formulation 

(paste and tablet) on parasite load and metabolic status in sheep. Forty 

five sheep, infected with various gastrointestinal parasites and kept on 

a grazing flock, were evenly divided into three groups. Sheep in the 

control group did not receive any treatment. The sheep in the remaining 

groups received the same dose of Oxyclozanid (5 mg /kg body weight) 

and Oxfendazole (13.5 mg/kg body weight) in combination either as 

tablet or as paste. Faeces and blood samples were collected pretreatment 

(day 0) and on day 14 post-treatment. Several gastrointestinal 

nematode parasites were detected in the faecal samples. Eggs per gram 

(EPG) on days 0 and 14 were 100±18 and 78±8 in CG, 96±12 and 6±3 

in TG, and 76±9 and 2±2 in the PG respectively. Serum total protein 

and globulin concentrations increased significantly (P

767 Effect of Zinc from Zinc Sulfate on Ewes’ Weight, Milk Yield, 

Zn Concentrations in Serum and Serum Alkaline Phosphates 

Activity of Varamini Ewes 

Ab. Zali, Ha. Amanlou, Al. Nik khah, Ah. Ahmad, Ka. Rezayazdi, 

Ma. Ganjkhanlou 

Animal Sciences Group, Animal Science, Karaj, Iran 

This experiment was conducted to investigate the effect of feeding 

supplemental zinc (zinc sulfate) in different levels (0, 15 and 30 mg/kg) 

on ewes weight, milk production, Zn concentrations in serum and 

serum alkaline phosphates activity. Thirty lactating Varaminni ewes 

were assigned to three experimental groups according to their live body 

weights, milk production and lambs sex in a completely randomized 

design. Ewes were fed a basal diet containing alfalfa, wheat straw, 

cottonseed meal, barley grain, wheat bran, cracked corn and vitaminmineral 

supplements at 3.2% of BW to meet NRC requirements for 

protein, energy, macro minerals and micro minerals. The basal diet 

contained 15 mg/kg Zn and Zinc sulfate was added to the basal diet to 

supply 30 or 45 mg/kg of dietary zinc. Milk yielded, milk composition 

and ewes’ weight was recorded at 7 and 21 days intervals respectively. 

Samples of the blood were taken three times (0, 35 and 70) for 

determination of Concentration of Zn, Cu and Fe, Na, K, Ca in serum. 

Also serum alkaline phosphates concentration of ewes was measured. 

Milk yield, milk composition and ewes’ weight of ewes were not 

affected by supplemental zinc (p>0.05). Alkaline phosphatase 

concentration was increased with supplemental zinc linearly and this 

increase was significant (p0.05). 

Key words: supplemental zinc, zinc sulfate, varaminni ewes 

768 Microscopic Comparison among Different Treatments Effects 

for Second Intention Skin Healing in Sheeps 

R. Godoy, JE. Barroso, FH. Ximenes, M. Castro, V. Mustafa, C. 

Leite, C. Pereira, L. Gouvea, L. Gontijo, JR. Borges 

Universidade de Brasília, FAV, Brasília-DF Brazil, Brazil 

Objective: Compare the effect of Stryphnodendron adstringens 

solution, Dakin’s solution, povidone-iodine and water for second 

intention skin healing in sheeps. 

Materials and methods: Four cutaneous wounds were made on both sides 

of thorax in 6 sheeps, with a circular “punch” of 0,8 cm of diameter, 

similar to the procedure for obtaining biopsies. Treatment began after 15 

hours and was maintained once daily. It was used sequentially, the 

treatments: T1. Water; T2. Stryphnodendron adstringens solution at 

10%, T3. Dakin’s solution (NaClO 0.5%) and T4. Povidone-iodine 

(PVPI) 0.1%. The biopsies, performed in the right and left sides, on days 

7 (D7) and 14 (D14), respectively, after the production of wounds, were 

sent for histopathologic analysis that evaluated swelling, exudate of 

polymorphonuclear (PMN), vascularization, mononuclear infiltrate, 

fibrosis, ulcer and reepitelization. The measurement standard of the 

factors cited were: Degree 0 = Absence; Degree 1 = Mild, Degree 2 = 

Moderate, Degree 3 = Intense. 

Results: The biopsies performed in D7 showed that the wounds treated 

with Dakin’s solution and Barbatiman’s solution showed degree 1 of 

swelling, the water treated, degree 2, followed by mild to moderate in 

the treatment with PVPI. Regarding fibrosis, T1 and T2 showed degree 

1 and in the others treatments mild to moderate degree were observed. 

The others parameters evaluated did not differ between treatments, 

with degree 3 to PMN exudate and ulcer, degree 1 to vascularization 

and absence to mononuclear infiltrate. In D14, the degree of swelling in 

the T1 and T3 was 0, in the T4 ranged from absence to mild and 

resulted in degree 1 in T2. Regarding PMN exudate, T3 resulted in 

degrre 0, in T1 and T4 was predominantly degree 1 and in T2 degree 3 

were measured. The vascularization in T2 showed degree 1 and in the 

others treatments were observed degree 2. The mononuclear infiltration 

was evalueted as degree 2 in T1 and as degree 1 in the others 

treatments. In T2, the degree of fibrosis observed was 1, in the T1 and 

T4 it were similar, presenting degree 2, and T3 ranged from moderate 

to intense. With regard to ulcer it obtained degree 0 in T3 and T4, 

degree 1 in T1 and degree 2 in T2. For reepitelization, T3 and T4 

resulted in degree 3, followed by moderate to intense degree to T1 and 

T2 with degree 1. Conclusion: Second intention skin healing in sheeps 

was more efficient in water and Dakin’s solution treatments. 

Key words: Stryphnodendron adstringens, histopathology, ruminant, 

wound 

769 Echodopplercardiographic Evaluation in Santa Ines Sheeps 

JE. Barroso, M. Mendes, CE. Silva, R. Godoy, JR. Borges 


Objective: Evaluate the cardiac size and function in Santa Ines adult 

sheeps to establish physiologic reference values. 

Materials and Methods: 30 sheeps not pregnant, aged between one 

and five years and average weight of 34.8 ± 7.41 kg were used. In right 

lateral decubitus and with a 2-4 MHz bandwidth microconvex 

transducer, in M-Mode, it was measured in cm: in diastole, the 

thickness of right ventricle wall (RVWd), of interventricular septum 

(VSd), of left ventricular wall (LVWd), diameter of right ventricle 

(RVd) and of left ventricle (LVd), and in systole, thickness of the 

interventricular septum (VSs), of left ventricle wall (LVWs) and left 

ventricle diameter (LVs). Diameters of systolic left atrium (LA) and 

diastolic aorta (AO) were measured in cm and the left atrium/aorta ratio 

(LA/AO) was established. Shortening fraction (SF%), was calculated 

with the formula (LVd-LVs/LVd)x100. Pulmonary (VPul) and aortic 

(VAO) velocity flow were measured in m/s and pulmonary (PGPul) 

and aortic (PGAO) pressure gradients in mmHg. It was evaluated 

pulmonary and aortic regurgitation by color flow mapping. Right 

parasternal long-axis left ventricular outflow view was used to obtain 

RVWd, RVd, LVd, LVs, LVWs, LVWd, VSs and VSd values. The 

same view was used on the heart base to get LA and AO values. VPul 

and VAO were obtained, with pulsed Doppler, in the right parasternal 

oblique left ventricular right ventricular outflow view and the left 

parasternal apical view, respectively. 

Results: The average values found for RVd and LVd were 1.34±0.46 

and 3.28±0.38, respectively, maintaining a 1 to 3 approximately ratio. 

RVWd (0.39±0.09) was half of LVWd value (0.62±0.12) and the 

LVWs, LVs, VSd and VSs average dimensions were 1.06±0.2, 

2,19±0.29, 0.92±0.12 and 1.18±0.15 respectively. FS found was 

32.96±6.19. The LA diameter (2.78±0.29) was similar to AO 

(2.44±0.27), and LA/AO ratio was equal to 1.15. VAO (0.85±0.12) and 

VPul (0.85±0.1) values were similar, as well as and PGPul (2.95±0.71) 

and PGAO (2.93±0,82). Discreet aortic and pulmonary regurgitation 

with low velocity and short duration were observed in 13% and 50% of 

the examined animals, respectively. 

Conclusion: This study establishes echodopplercardiographycs 

reference values to Santa Ines sheeps. Others parameters, as well as 

different breeds, should be evaluated. 

Key words: ovine, cardiology, cardiac size, cardiac function 

770 Aspects Regarding Maedi Visna Diseases Incidence and 

Prevalence in Covasna County 

R. Popescu1 , C. Horhogea2 , B. Siko1 1 Regional Veterinary Laboratory, Virology, Sfintu Gheorghe, 

Romania 

2 University of Agricol Sciences and Veterinary Medicine, Veterinary 

Medicine, Iasi, Romania 

Objectives of study: The researches were made to establish the 

incidence and prevalence of Maedi Visna infection in Covasna County. 

Materials and Methods: The researches were made on 8300 samples 

from Covasna County sheeps, with different owners during 2003-2007. 

For the serological exams the folowing ELISA kits were used: 

CHEKIT CAEV/MVV Screening, IDEXX Lab., Caprine Arthritis- 

Encephalitis Virus Antibody Test Kit, VMRD Inc., ELISA MAEDI- 

VISNA/CAEV Serum, Pourquier Institut, Serological Diagnosis of 

Maedi-Visna By testing for Gp135 antibodies in serum using the agargel 

immunodiffusion method, Pourquire Insitut. 

Results: In 2003 were tested 2019 serum samples through ELISA, 405 

being positive. In 2004 were tested 830 samples revealing the fact that 

193 were positive, representing a prevalence of 23%. 

In 2005 were tested 1545 samples, 390 being positive, and revealing a 

prevalence of 25%. In 2006 were tested 1140 serum samples,843 being 

negative and 297 positive showing a prevalence of 25%.The tests 

revealed the fact that in 2006 the diseases was extended in all 38 

circumscriptions of the Covasna County. In 2007 the serological tests 

executed on 1582 samples showed that 1107 samples were negative 

and 475 were positive, indicating a prevalence of 30%. 

Conclusions: The researches made between 2003-2007 showed 

variations of the Maedi Visna infections incidence and prevalence. It 

was observed that the diseases incidence in 2004 was of 84 positive 

cases, in 2005 was revealed an increasing till 194 positive cases, in 


2006 was observed a small decreasing of the incidence till 166 positive 

cases, and in 2007 a new increasing till 275 positive cases. The 

prevalence compared to the serological exams made each year revealed 

in 2003 405 positive samples from the 2019 tested, in 2004 from the 

830 samples 193 were positive, in 2005 from the 1545 samples 390 

were positive, in 2006 297 positive samples from 1140,and in 2007, 

from 1582 serological exams 475 were positive. If in 2003 antibodies 

against Maedi Visna virus were detected in sheeps from 25 Sanitar 

Veterinary Circumscriptions, their number increased each year, in 

2006, 2007 antibodies being detected in all 38 circumscriptions of the 

Covasna County. 

771 Effect of Intracervical Infusion of Prostaglandin F 2a and 

Prostaglandin E2 on Dilation of Cervix in Ewes with 

Ringwomb 

M. Eslampour 1 , P. Tajik 1 , S. Bokaie 2 , H. Moaddab 1 , A. Moghiseh 1 , 

B. Khoramian 1 , S. Rafia 1 , E. Banihasan 1 

1 University of Tehran, Dept. of Clinical Sciences, Tehran, Iran 

2 University of Tehran, Department of Food Hygiene, Tehran, Iran 

To evaluate the clinical efficacy of administration of prostaglandin E2 

(PGE 2) and prostaglandin F 2a (PGF 2a) intracervically by comparing 

these with digital manipulation on dilation of cervices in ewes with 

ringwomb .Pregnant ewes which cervices failed to dilate during 

parturition assigned randomly to three treatment groups: 1) 7 ewes 

manipulated their cervices digitally for ten minutes and if this effort 

had not responded, after ten minutes another effort initiated, 2) 6 ewes 

received 1 mg infusion of PGF 2a (Lutalyse; Pfizer Animal Health) 

dissolved in 2 ml of 30% gelatine (Sigma-Aldrich Co.) intracervically 

and, 3) 6 ewes received 0.5 mg infusion of PGE 2 (Prepidil; Pfizer 

Health) intracervically . For infusion of PG 2a, a speculum introduced 

into the vagina so that the external os cervix could be seen in the light 

of the speculum lamp. The tip sterile pipette was inserted about 2 cm 

into the cervix and the gels deposited in to the lumen of cervices. For 

prevention of leakage of PG 2a gels required restraining ewes in to the 

lateral recumbency for 15 minutes after infused. The response to the 

treatment would be assigned positive if proceeding on parturition and 

delivery of live lambs was occurred one hour after administration of 

treatment. All of the ewes that did not response to the treatment, 

monitoring for two hours later and if there was not any proceeding on 

parturition they would be culled consequently. Results showed that 

ewes were treated with PGE 2a interacervically had greatest response 

compared with treatment with digital manipulation and intracervical 

infusion of PGF 2a (5/6s vs. 3/7 and 3/6 respectively). However, 

response to treatment did not differ between the groups significantly 

(p>0.05). Culling rates from three groups revealed that ewes in digital 

manipulation group had greater culling rates compared with PGF 2a and 

PGE 2a groups (4/7 vs. 2/7 and 1/7 respectively, p>0.05). It can be 

concluded that using PGE 2 in the ewes with undilated cervices after 

initiated of abdominal straining can be an effective treatment and 

preventive approach for culling of pregnant ewes. 

772 Influence of Reduced Dietary Crude Protein Level on 

Performance of Dairy Cows under Heat Stress 

A. Asadi, G. Ghorbani 

Isfahan University of Technology, Department of Animal Science, 

Isfahan, Iran 

Two groups of high yielding early lactation cows were allocated to 

either high (17% DM, 106 cows) or low (15.6% DM, 102 cows) crude 

protein diet. Mean DIM, milk yield and parity for cows in high and low 

protein treatments were; 59.5±21 and 65.3±26d; 38 and 37.3 kg/d; 3.1 

and 2.9, respectively. Mean daily ambient temperature was 37.3 ˚C 

throughout the trial. Cows were in the trial for six weeks allowing two 

weeks for adaptation and the remaining 28 days for data collection. Dry 

matter intake and milk production were recorded daily and diets and 

milk were sampled weekly. MIXED procedure was used for statistical 

analysis including time of sampling in the repeated statement and 

assuming cow as random effect. Reducing dietary protein resulted in 

lower dry matter intake (22.4 vs. 24.9 kg/d, P

775 Preventive Medicine and Health Problems in Chios Breed 

Flocks of Sheep 

A. Gelasakis, G. Valergakis, G. Arsenos, P. Fortomaris, G. Banos 

Aristotle University of Thessaloniki, Animal Production, Thessaloniki, 

Greece 

The objective of the study was to assess the most frequent health 

problems and the main preventive measures implemented in 

intensively and semi-intensively reared flocks of Chios dairy breed of 

sheep in Greece. A total of 67 flocks of the Chios Sheep Breeders 

Cooperative Macedonia were involved in the study. The research 

comprised farm visits by the same veterinarian who used a 

questionnaire designed for in depth interviews with farmers. The 

questionnaire involved questions about livestock, facilities, equipment, 

management practices, nutrition, and major health problems. Data were 

analysed using methods of descriptive and analytical statistics. Results 

revealed that a significant percentage of farmers vaccinated against 

Clostridial diseases, Contagious agalactia and Enzootic abortion 

(100%, 79% and 75%, respectively). On the contrary, fewer farmers 

vaccinated against Pasteurellosis, Gangrenous mastitis and 

Paratuberculosis (47.8%, 20.9% and 4.5%, respectively). The 

preventive use of Vitamin-E and Selenium in newborn lambs was a 

common practice in 66 flocks. All the farmers used anthelminthic 

treatments but with different intensity; 17.9% of the farmers used it 

once per year, 50.7% twice and 31.4% three times per year. Diarrhoea 

(>10%) and pneumonia (>5%) in lambs was observed in 33 and 9 

flocks, respectively. A serious problem in ewes was mastitis; in 20.9% 

of flocks the incidence was >10% and 50.7% of the farmers used 

intramammary infusion of antibiotic as a preventive measure. 

Lameness and abortion were also significant problems (in 40.3% and 

23.9% of the farms, respectively. they were >5%). For the prevention 

of lameness, 18 farmers performed programmed foot trimming and 5 

foot bathing, whilst 6 and 1 added ZnO and Biotin to the ration. The 

results showed also a significant negative correlation (P

for O69 strain), and clinical signs were not obvious, mice dead 

suddenly. The lesion profile identified is close to that of other scrapie 

strains profile and clearly different from conventional BSE one. The 

PrPsc was detected in several areas of the mice brains as diffused and 

granular aggregates and more specifically into the thalamus, the 

cerebellum and the choroid plexus. The GFAP detection has revealed a 

strong astrocytosis. Furthermore, hyperchromatic neurones were 

observed in ‘TAC’ scrapie infected mice indicating a severe 

neurodegeneration. Finally, the PrPsc molecular profiles have 

confirmed that the ‘TAC’ isolate is a classical scrapie strain. 

Key words: scrapie, sheep, prion, strain typing, transgenic mice 

779 Orchitis and Epididymitis in Ovine Caused by Histophilus 

somni 

I. Enriquez 1 , V. Tenorio 1 , F. Morales 1 , E. Diaz 1 , S. Nieto 1 , 

F. Aguilar 1,2 

1 INIFAP, Bacteriologia, México DF, Mexico 

1 Facultad de Medicina Veterinaria y Zootecnia - UNAM, 

Microbiology and Immunology, México DF, Mexico 

Reproductive tract infections are quite important for economics in 

ovine production. Within these, ram infectious orchiepididymitis (RIO) 

is widely spread, with prevalence up to 50 % of the rams of a flock. 

Because of this, when any sign of the presence of this kind of infection 

is seen, action must be taken immediately. The objective of this study 

was to determine the etiology of a RIO outbreak in a fattening facility 

for Pelibuey sheep, with approximately 3,000 adult rams. Sixty cases of 

unilateral orchitis and epididymitis were detected by clinical 

examination. The affected males were separated and confined into 

another pen in order to avoid greater dissemination of the problem. 

Samples from testicle and epididymis form 4 (4/60) affected rams, 10 

(10/60) preputial swabs and 30 (30/60) blood sera were obtained to 

carry out bacteriological, serological and histopathological studies. 

Bacteriological studies were done to the tissue samples and the swabs, 

identifying the isolations by the Polymerase Chain Reaction technique 

(PCR). The Double Agar-Gel Immunodiffusion test was used for the 

serological studies with antigens specific for H. somni, A. seminis and 

B. ovis. H. somni was isolated from the 4 (100%) testicles, as well as 

from the 4 (100%) epididymides from the 4 animals to which were 

submitted to castration. Regarding the serological tests, 12/30 (40%) 

sera were positive to H. somni, including the 4 animals with positive 

isolations; one serum resulted positive for B. ovis as well as for H. 

somni. Severe multifocal abscessed fibrosis, azoospermia and necrotic 

areas that corresponded to a suppurative process with active-chronic 

course were found in the histological study of the 4 testicles and 

epididymides along with cystic fibrosis, compatible with bacterial 

infection. No evidence of another specific pathological process was 

found. It is concluded that in the presentation of the RIO outbreak was 

evident the role of H. somni. There are no previous reports in Mexico 

of a similar case. The coexistence situation among rams that favor 

mounting between themselves might have had an important effect on 

the outbreak presentation. 

Partially financed by the SAGARPA-CONACYT # 4216361P. 

780 Importance of Percentage Participation by Insume in total 

Production Cost to Sheep Production 

G. Gil, A. Alonso, V. Espinosa, F. Alonso 

UNAM, Departamento de Economía, Administración y Desarrollo 

Rural, MEXICO, D.F., Mexico 

The objective of this study was determinate percentage participation in 

total cost of every insume in productive process of a weaning lamb and 

one Kg. of fatten lamb for supply.The study was made in Maravatio, 

Michoacan, Mexico, with 11 productive units. Semi-structured 

interviews, questionnaires application, direct observation and actionparticipative 

method were the instruments to get the information. 

Results showed that producers had an average of 47 sheeps and 33 

weaning lambs by flock. The productive cycle lasted one year. The total 

cost on average of a weaning lamb was $ 18 324.14, variable costs 

represented the 70% of total cost and fixed costs represented the 30% 

of total cost. The average of cost production was $ 642.92, feed took up 

the first place with a 54.42%, followed in second place by the 

exhaustion of reproductive animals, that took up a 22.82%. The rest of 

the insumes followed in a descended way. The productive units have an 

average production of 459.36 Kg. of lamb of living weight by cycle, 


which lasted 88.36 days on average. The quantity of lambs in every 

fatten batch was 10.64 on average, and each one got a weight of 43.21 

Kg for sale. The total cost on average of 1 kg. of supply living lamb was 

$ 32 907.14, variable costs represented the 98% of total cost and fixed 

costs represented the 2%. The average of production cost was $ 20.95, 

the first place was taken up by the animals, with the 70.48%, the second 

place was taken up by the feed with the 24.44%. The rest of the insumes 

followed in a descended way. The pay by Kg of living lamb is $ 23.00, 

that’s the reason that, although the sale price is highest than their 

production cost and get some profits, incomes get by the activity are not 

enough to cover their basic neededs. It is important to identify 

deficiencies in the productive system. To considerate sheep production 

as an developed option in rural communities and be competitives, is 

important to make it with low costs adopting technologies that lend the 

producers bring the commercial aperture in the best way. In conclusion, 

sheep production is a good alternative in studied units. With the 

knowledge of the main insumes in total cost it could be possible to 

adopt strategies and politics to reduce the production cost. There are 

many rural families that get their sustenance from this activity, so is 

important to do more researches about this aspect. 

Key words: production cost in sheep, lamb 

781 Evaluation of Three Antigens for Diagnostic of Brucella ovis 

by ELISA 

I. Tuxpan 1 , L. Hernandez 1 , J. Acosta 1 , L. Ontiveros 1 , J. Martinez 2 

1 INIFAP, Bacteriologia, México DF, Mexico 


Departamento de Medicina Veterinária Preventiva, México DF, 

Mexico 

Brucella ovis is a specific cause of epididymitis in rams. Diagnosis of 

infections in flocks is based on scrotal palpation, semen culture and 

serological testing. Three antigen extracts were evaluated in an indirect 

enzyme-linked-immunosorbent assay (ELISA) for detection of 

antibodies to Brucella ovis in rams. B. ovis Reo 198 was used as 

reference strain to obtain the different antigens. The antigens used 

were: a) soluble extract; b) antigen obtained by extraction with heat and 

c) soluble extract ultracentrifuged. The total proteins were 

determinated by the Bradford method. All reagents were standardized 

by checker board titration and saturation of plates with the selected 

antigen. The negative samples control serum were obtained from 36 B. 

ovis free animals, and the positive control sera were from 36 animals 

with clinical epididymitis and positive isolate of B. ovis and 36 positive 

samples serum to B. melitensis. The cutting point was 1.2 for the three 

antigens. For the antigen obtained by heat extraction the sensitivity was 

88.89% and specificity of 97.22%, predictive value positive was of 

96.97 and predictive value negative of 80.22. The extract 

ultracentrifuged soluble antigen had a sensitivity of 86.11% and 

specificity of 97.22%, predictive value positive 96.88 and predictive 

value negative of 87.5. Extracted soluble had the 75% and 97.22% of 

sensitivity and specificity respectively, predictive value positive of 

96.43 and predictive value negative of 79.55. No cross reaction was 

found with serum sample from animals infected with B. melitensis. The 

best antigen was the heat extract. 

Key words: Brucella ovis, epididymitis, ELISA, rams 

782 Experimental Infection of Ovines with Histophilus somni 

Isolations from Several Sources 

E. Diaz-Aparicio 1 , F. Aguilar 1,2 , B. Arellano 2 , I. Enriquez 1 , 

V. Tenorio 1 

1 INIFAP, Bacteriologia, Mexico, D.F., Mexico 


Microbiology and Immunology, Mexico, D.F., Mexico 

The purpose of this study was to establish if there is a difference in 

virulence when infecting ovines experimentally with Histophilus somni 

isolates: a) 2336 bovine pneumonic lung origin strain which was 

previously inoculated in an ovine and recovered; b) an isolate from 

orchitis and epididymitis; c) an isolate from the brain of an ovine with 

nervous signs; d) an isolate from vaginal exudate of a clinically healthy 

young ovine. A total of 25 male sheep were divided in groups of five 

animals each, which were inoculated with 1x10 7 CFU/ml in the 

epididymis, and a control group which was inoculated with 

physiological saline solution. Clinical inspection was carried out 

everyday, and one month after the challenge the animals were

slaughtered observing macroscopic and microscopic lesions. 

Epididymitis was reproduced in the experimentally infected ovines, but 

H. somni was only isolated from ovine groups inoculated with isolates 

originating from epididymitis and semen. The group that had greater 

lesions of epididymitis was that which was inoculated with the isolate 

originating from the epididymitis outbreak, followed by the group 

inoculated with the isolate originating from vaginal exudates. The 

groups inoculated with the isolate from brain and strain 2336 showed a 

lesserdegree of epididymitis. It was demonstrated that there is a 

difference in experimental infection capacity between isolates of H. 

somni from different origins as epididymitis occurred and the bacteria 

was recovered only in groups of sheep inoculated with isolates 

originating from epididymitis and vaginal exudate. 

This work was partially funded by project 11928 SAGARPA- 

CONACyT, Mexico 

783 Umbilical Cord Healing: Comparative Study of Use of 

Chlorhexidine, Iodine, and Picric Acid/Dichlorvos/Phenol* 

Solutions in Sheep 

L. Mendes, A. Lombardi, F. Feres, J. Peiro, F. Feitosa, F. Cadioli, 

R. Lataro, S. Perri 

Sao Paulo State University - Aracatuba Campus, DCCRA, Aracatuba, 

Brazil 

The aim of this study was to compare the efficacy of several antiseptic 

solutions used for umbilical care of newborn sheep.Thirty-five lambs, 

cross-bred, born between April and June 2007, both sexes, were 

monitored from birth until the he umbilical cord fell off. Animals were 

randomly divided into 3 groups of 10 lambs each, and a control group 

(n=5), and were kept on pasture with their mothers. From birth until 18 

hours of life, lambs were weighed and the umbilical cord was cut at 3 cm 

from the abdominal wall and received one of the following treatments: 

dipping with 0.5% chlorhexidine, dipping with 2% iodine or cleansing 

with 2% picric acid/dichlorvos/phenol*, and the control group was not 

treated. Animals were checked daily for the presence of maggots, and 

excessive secretion or humidity of the umbilical cord. After umbilical 

cord falling off, lambs were weighed and the body weight gain was 

calculated. Umbilical fall off times and body weight gain data were 

submitted to ANOVA followed by Tukey´s test for multiple comparisons 

of means among groups (SAS v.8, p < 0.05%). Umbilical fall off times 

were not different among groups. However, umbilical cords of lambs 

treated with 2% picric acid/dichlorvos/phenol* gained more weight while 

controls showed a worse performance in gaining weight. Lambs treated 

with chlorhexidine or iodine did not show statistical differences between 

them, but were different from the other 2 groups. The season of the year 

may have interfered with the results of this study, as the dry season of the 

year visually interfered with the umbilical retraction and fall off time. By 

this reason, the control group was not different from the others. Although 

the 2% picric acid /dichlorvos/phenol* or iodine groups did not show 

differences in umbilical fall off time, it was observed that animals which 

received these solutions had a higher weight gain as compared with 

controls. We concluded that more studies during the raining season should 

be done to attempt to confirm the interference of the season of the year 

with the umbilical fall off time and body weight gain. 

*Pecuarista D’oeste - Brazil 

Key words: navel, sheep, lambs 

784 Comparative Pharmacokinetic Profiles of Moxidectin in 

Sheep following Administration of a Conventional or Long- 

Acting Injectable Moxidectin Formulation 

R. DeLay 1 , F. Blond 2 , E. Lacoste 3 , F. Bassissi 3 , D. Rock 1 

1 Fort Dodge Animal Health, R&D, Princeton, United States 

2 Fort Dodge Animal Health, R&D, Tours, France 

3 Avogadro, R&D, Parc de Genibrat, France 

Moxidectin is a macrocylic lactone compound with broad spectrum 

activity against many internal and external parasites of animals. 

CYDECTIN ® (1% moxidectin) Injectable, an aqueous-based product, 

has been used for parasite control in sheep. Recently, an oil-based 

formulation containing 2% moxidectin has demonstrated long-acting 

control of parasites in sheep. The objective of this study was to compare 

the pharmacokinetic profiles of moxidectin following administration of 

these two formulations in sheep. Twelve young male sheep 

(Tarascon/Tarascon crosses) averaging 24 kg b.w. were divided into two 

similar groups. Group 1 received CYDECTIN (1%) by subcutaneous 

injection in the neck to provide 0.2 mg moxidectin/kg b.w. Group 2 

received moxidectin LA by subcutaneous injection at the base of the ear 

to provide 1 mg moxidectin/kg b.w. Blood samples were collected 

periodically for up to 80 days and 161 days post-treatment for Groups 1 

and 2, respectively. Plasma was haarvested and analyzed for moxidectin 

using an HPLC method validated to 0.5 ng/mL. The moxidectin plasma 

concentrations over time were subjected to non-compartmental 

pharmacokinetic calculations. The C max for Groups 1 and 2 were not 

significantly different (32.3 ± 7.2 vs. 23.6 ± 6.7 ng/mL; P 6 mm; PO), ovulated (OV) and corpora 

lutea (CL). Then, the females were allocated in three periods. In period 

1 (P1), those that ovulated before the first evaluation. In period 2 (P2), 

those that ovulated among the 42 and 48 h, and period 3 (P3), the ones 

that ovulated after 48 h. The data were analyzed by ANOVA being 

them means compared using Kruskal-Wallis test (P0.05). In conclusion, the dates suggested that the addition of LH to 

the protocol promoted tendency of anticipation of the ovulations. 

Key words: superovulation protocols, ovulation inducer, ovine 

786 Diagnostics of Toxoplasma gondii by PCR in Sheep of Flocks 

from Eastern Slovakia 

L. Luptakova 1 , P. Balent 1 , A. Valencakova 1 , M. Huska 2 , 

E. Petrovova 1 

1 University of Veterinary Medicine, Department of Biology and 

Genetics, Kosice, Slovakia 


2 University of Veterinary Medicine, Clinic of Pigs, Kosice, Slovakia 

The study presents the results of examination of 62 sheep from 3 flocks 

of eastern Slovakia for the presence of Toxoplasma gondii antibodies 

by complement fixation test. 13 samples were found to be positive at 

the titre of 1:64, 10 were positive at 1:128 and two at 1:256 dilution. 

Subsequently, DNA was isolated from blood samples of these animals. 

We then used PCR analysis with specific primers in search for T. gondii 

DNA. In PCR analysis, 5 of 25 (20 %) of samples tested positive. 

Key words: complement fixation reaction, polymerase chain reaction, 

sheep, Toxoplasma gondii 

787 Relationship between Physical Exam of the Mammary Gland 

and Cellularity of Milk in Santa Ines Sheep 

M. Blagitz 1 , V. Gomes 1 , N. Benites 2 , P. Melville 2 , M. Azedo 1 , 

C. Batista 1 , A. Della Libera 1 


1- Department of Clinical Medicine, Sâo Paulo, Brazil 


animal science department, Sâo Paulo, Brazil 

To compare milk cellularity of Santa Ines sheep and alterations in 

udder physical exam, udders of 292 ewes raised in Sao Paulo were 

submitted to physical exam and Tamis exam. Milk samples were 

collected for bacteriological exam, CMT, microscopic SCC and 

automatic SCC. Udders were ranked by results of physical exam: those 

without alterations both in inspection and palpation (Rank 0); those 

with alterations in inspection (Rank 1); those with alterations in 

palpation (Rank 2); and those with alterations both in inspection and 

palpation (Rank 3). Results showed that udders rank 3 had more 

alterations in the Tamis exam than those rank 0 (p

sex evaluation and seven animals of Morada Nova (MND), eight of 

Rabo Largo (RLD) and seven of Santa Ines (SID) were used for the 

breed variable. The lambs were weighed weekly and analyzed in the 

following periods: at birth, in the weaning period, 30 and 60 days post 

weaning. Blood samples were obtained by jugular venipuncture and 

stored in tubes containing ethylenediaminetetraacetic acid (ETDA) 

and then the erythrogram was performed. The values obtained for 

weight, the number of red blood cells, the packed cell volume and the 

hemoglobin concentration were statistically significant (p

794 Serum Haptoglobin and Fibrinogen Levels along the 

Pregnancy and Post-partum of Ewes 

B. Bastos, C. Caribe Ayres, U. Burghgrave, A. Ferreira, T. Pires, 

A. Gusmao, J. Guimaraes 

Escola de Medicina Veterinária UFBA, Salvador - Bahia - Brasil, 

Brazil 

The increase in serum acute phase proteins is stimulated by traumatic, 

inflammatory or infecctious events. However, studies have 

demonstrated the presence of haptoglobin (Hp) and fibrinogen (Fb) in 

mammalian reproductive tissues in the abscence of inflammation, 

indicating a potential role for these proteins in reproductive process. 

Considering that no information has been published concerning the 

values of acute phase proteins along the pregnancy of small ruminants, 

this work aimed to establish the dynamics of Hp and Fb along the 

pregnancy and post-partum of ewes. Fifty clinically health animals of 

Santa Ines breed were selected in a commercial flock from Bahia, 

Brazil, and blood was sampled prior to the oestrus synchronization and 

artificial insemination. Thirty-eight pregnancies were diagnosed and 

blood was collected every 14 days until post-partum. Serum Hp was 

determined based on its hemoglobin-binding capacity and plasma Fb 

through the heat precipitation technique. The presence of any possible 

infectious process was assessed through white cell count. Data were 

analyzed in SPSS v.13 statistical program through the non-parametric 

Wilcoxon test. Concerning the Hp results, significant higher values 

(p

post translational modifications. We found no putative coil-coiled 

structure in ovine TLR7 protein by 14, 21 and 28 windows analysis. 

The putative structural analysis was performed by TMHMM as well as 

PFAM servers, wherein we found that the sheep TLR7 ectodomain 

starts at amino acid position (AA) 27 (LRR N terminus 27 to 60 AA) 

and terminates at 831 (LRR C terminus 808 to 831 AA). PFAM 

showed 5 LRRs in ectodomain lied between 222-244, 311-334, 648- 

670, 697-718 and 745 to 767 AA with resemblance of super-repeat 

motifs. We predict that these regions are the most probable functional 

regions involved in PAMPs recognition. The other variable or 

conservative forms of LRRs of ovine TLR7 are not yet predicted, 

however, we suppose their presence in the ectodomain. The 

transmembrane domain spanned between 838 and 866 according to 

TMHMM probability prediction and the TIR domain spanned between 

890 and 1029. This study makes a benchmarking for further ovine 

TLR7 gene analysis like SNPs detection, protein structural and 

functional analysis. 

Key words: TLR7, sheep, LRR, PAMP, toll like receptor 

798 Drug Interaction of Succinylcholine and Ivermectin in Sheep 

A. Raayat Jahromi, N. Vesal, S. Nazifi 

School of Veterinary Medicine, Shiraz Univrsity, Shiraz, Iran, 

Clinical Sciences, Shiraz, Iran 

Drug interactions subject is one of the most important aspects of 

therapy when two or more drugs are administered concurrently. In a 

former study, an unusual reaction to succinylcholine was observed 

following ivermectin administration. The aim of this study was to 

evaluate possible interaction between these drugs. Ten native male 

adult sheep, about one year old, were randomly assigned to control and 

treatment groups (n=5). In order to study the possible effect of 

ivermectin on plasma cholinesterase activity, measurement of serum 

enzyme activity was performed by manual and automated methods 

before ivermectin (0.02 mg/kg, SC) and normal saline (as placebo, SC, 

in control group) injection at following intervals: 72 hours before 

anesthesia, immediately before anesthesia, 1, 3, 5 and 15 days after 

anesthesia. Following premedication with xylazine, anesthesia was 

induced with ketamine and maintained with halothane in oxygen. After 

stable anesthesia, succinylcholine (0.1 mg/kg) was administered 

intravenously. Temperature and cardiopulmonary parameters (heart 

rate, respiratory rate and end-tidal PCO 2) were recorded during 

anesthesia at 5 minutes interval. Time from succinylcholine injection to 

apnea, depression of twitch reflex, recovery times (return of 

swallowing reflex, spontaneous breathing and voluntary 

movements)and times to sternal and standing position were measured 

in both groups. Data analysis was done by one-way ANOVA and 

independent t-test and values of P0.05) although sheep fed with 0.1 

% enzyme showed a little higher plasma glucose concentration than sheep 

fed with 0.05 % enzyme (p0.05). 

Result: This research showed that enzyme usage in two levels in 

comparison with no enzyme application in diet has no effect on the 

plasma glucose level (P>0.05) although sheep fed with 0.1 % enzyme 

showed a little higher plasma glucose concentrations than sheep fed 

with 0.05 % enzyme (p0.05) and also the blood urea nitrogen 

concentration are not different (p>0.05) among treatments can be 

connected with that diets were isoenergetic and have similar protein 

percentages. Hristov et al. (1998) found that enzyme in diet of animal 

has no effect on the plasma glucose and blood urea nitrogen 

concentrations. 

Key words: NATUZYME, sheep nutrition, enzyme complex, plasma 

glucose, blood urea nitrogen (BUN) 

801 Effect of Feeding NATUZYM (a Multi Enzyme Complex) 

on Ruminal pH Level and Ammonia Nitrogen Concentration 

in Sheep 

H. Baghershah, K. Rezayazdi, M. Dehghan-banadaky, A. Zali, 

M. Khansefid 

University of Tehran, Animal Science, Karaj, Iran 

In this research Twenty-four Varamini male sheep (An Iranian 

vernacular breed) with initial birth weight of 22±1.5Kg were used to 

evaluate the effects of feeding an enzyme complex on the ruminal pH 

and ammonia nitrogen concentration in sheep. . The sheep were 

divided into three groups and fed with three diets: a diet without the 

enzyme (control diet) and diets contained 0.05% and 0.1% enzyme. All 

diets include 1:1 forage to concentrate ratio. . Enzyme added to 

concentrate and then mixed with forage in order to make a uniform diet. 

All Sheep were fed at adlibitum level. Diets were isoenergetic and 

contained similar protein percentages. The design of this research was 


complete random design (CRD). The research results show that 

ruminal pH and also ruminal ammonia nitrogen concentration have no 

statistical differences among treatments (p>0.05). 

Results: Ruminal pH levels and rumen ammonia nitrogen 

concentrations were not different (p>0.05) among treatments. 

Discussion: Reasons for that ruminal pH and rumen ammonia nitrogen 

concentration were not different (p>0.05) among treatments can be 

connected with that diets were isoenergetic and had similar protein 

percentages or equality of dry matter intake and also uniformity of 

nutrients digestibility. Some researchers found similar and some other 

researchers reached to different results from the present research 

consequences may be due to multi enzyme contents, enzyme 

application methods, enzyme types, enzyme levels, animal type or diet 

formulations which were apply with enzyme. 

Key words: NATUZYME, sheep nutrition, enzyme complex, ruminal 

pH, ammonia nitrogen 

802 Effects of DFM on Rumen Fermentation in Iranian Sheep 

H. Khazanehei 1 , K. Rezayazdi 1 , A. Nikkhah 1 , M. Riazabas 2 , H. 

Khalilvand 1 , N. Vahdani 1 , P. Hossein-Nia 3 , S. Ghovvati 4 

1 University of Tehran, Animal Nutrition, Karaj, Tehran, Iran 

2 University of Baghdad, Animal Nutrition, Baghdad, Iraq 

3 Isfahan University of Technology, Animal Science, Isfahan, Iran 

4 University of Mashhad, Animal Sciences, Mashhad, Iran 

A study was conducted to determine whether a direct-fed microbial 

(DFM) could be improvingruminal fermentation. Four Iranian sheep 

(Zandi breed, average BW 34+4.2 kg), fitted with ruminal cannulas, were 

used in a 2_2 cross-over design. Each experimental period was 21 days. 

Four rations were formulated to be iso-nitrogenus and iso-caloric and 

represent two sources of roughage with two levels of DFM (0 and 5 

gr/day/head). Ingredients of diets were alfalfa hay and corn silage (as 

different sources of roughage), wheat straw, barley grain, canola meal, 

Mineral-vitamin mix, calcium carbonate and salt. DFM was contained 

Enterococcus faecium, Lactobacillus acidophilus, Lactobacillus casaei, 

Bacillus subtilis (lactate producing bacteria) and Saccharomyces 

cerevisiae. DFM was added to control diet (TMR, 50% forage: 50% 

concentrate, dry matter basis) just before feeding. Ruminal fluid samples 

were taken from rumen at 0, 2, 4, 6 and 8 h to determine rumen pH and 

concentration of N-NH3, and at 4 h to determine concentration of VFA. 

Total VFA increased and mean ruminal ammonia decreased in diets with 

DFM compared to the control. DFM had no effect on propionate and 

acetate concentrations, acetate:propionate ratio, mean ruminal pH and 

maximum pH, however, decreased the nadir of ruminal pH. Increase in 

total VFA and decrease in mean rumen ammonia concentrations could be 

a result of higher microbial growth and activity in rumen of sheep that 

used DFM. The presence of lactate-producing bacteria is thought to help 

the ruminal microflora adapt to the presence of lactic acid and decreased 

the drop in ruminal pH following meals in sheep that used DFM (Table 1). 

Table 1. Characteristics of ruminal fermentation in sheep fed DFM 

Item 1 2 3 4 SE 

pH – – – – – 

Mean 6.49 6.55 6.45 6.51 0.07 

Min 5.85 b 5.93 b 6.04 a 6.1 a 0.04 

Max 7.05 7.12 6.98 7.06 0.04 

VFA 

Total, mM 71.26 d 82.3 b 75.91 c 88.6 a 3.42 

Acetate (A) % 37.69 b 41.97 a 38.21 b 43.63 a 1.5 

Propionat (P) % 19.46 21.96 20.23 22.87 1.2 

A:P 1.93 1.91 1.88 1.9 0.63 

NH 3 N mg/l 

Mean 8.47 bc 9 a 7.65 d 8.34 c 0.1 

a, b, c dMeans within a row that do not have a common superscript are different 

(p< 0.05), 1: control contain corn silage as a source of roughage, 2: control contain 

alfalfa hay as a source of roughage, 3: diet 1 with DFM, 4: diet 2 with DFM. 

Key words: DFM, alfalfa, corn silage, rumen fermentation 

803 Effect of Season and Photoperiod on the Time of First 

Postpartum Ovulation in Awassi Ewes 

V. Faigl 1 , M. Keresztes 1 , A. Márton 2 , H. Fébel 3 , M. Kulcsár 1 , 

S. Cseh 1 , L. Solti 1 , Gy. Huszenicza 1 


1 Szent István University Faculty of Veterinary Science, Departament 

of Animal Reproduction, Budapest, Hungary 

2 Georgikon Faculty of Agriculture, University of Pannonia, 

Keszthely, Hungary 

3 Research Institution of Animal Breeding and Nutrition, 

Herceghalom, Hungary 

Our aim was to study the influence of season and photoperiod on the 

time of first postpartum (pp) ovulation in non-suckling, dairy ewes. 

Trials were conducted in a commercial Awassi flock, where lambs are 

weaned immediately after birth. In the first experiment (Exp.1.) autumn 

lambing (AL; n=27) and spring lambing (SL; n=38), 1-11 parity ewes 

were used. Milk progesterone (P4) was determined trice weekly 

between d 7 and 110 pp. The time of the first ovulation was estimated 

by means of individual P4 profiles. Blood samples were collected 1 

week before lambing and again on week 1, 2 and 5 pp and assayed for 

some metabolites (non-esterified fatty acid, ‚OH-butyrate) and 

metabolic hormones (insulin, IGF-I, thyroxin). High proportion (89%) 

of AL dams ovulated before d 35 and became cyclic thereafter. 41% of 

them became pregnant from the 1-4 th ovulation. The non-conceiving 

ewes (n=16) became acyclic by Jan-Febr. Incidence of persistent 

corpus luteum (CLP; n=5) and short luteal phases (sCL; n=8; CLP and 

sCL together in 4 cases) was frequent among non-conceiving dams. In 

contrast only 39% of the SL ewes ovulated before d70 pp. P4 levels 

during luteal phase in cyclic animals were lower, and length of cycle 

was longer in SL compared to AL. No CLP or sCL was detected in SL. 

61% of SL remained acyclic till the end of trial. Plasma metabolits and 

metabolic hormones were in physiological range in the periparturian 

period. In Exp. 2. 48 autumn-lambing, 2-7 parity ewes were used. 

Animals were housed in open sheds. In mid September (1 week before 

the expected date of parturition) ewes were allotted into two treatment 

groups with regard to equal distribution of age, lambing date and 

former milk yield. Long-day photoperiod (LD) group (n=23) was 

exposed to artificial light from sunset till midnight (approx 16 hours 

light/8 hours dark). Control group (n=25) received no treatment 

(natural photoperiod). Sampling protocol was similar to Exp. 1. The 

time of first pp ovulation tended to delay in LD animals compared to 

Control (average 25.87±1.63 vs 21.5±1.72 days pp; surv. analysis 

P=0.093). Metabolic parameters remained between normal ranges 

throughout the experiment. In conclusion: i) ovarian function of awassi 

population became seasonal under temperate continental weather. ii) 

First pp ovulation of non-suckling, autumn-lambing dams may happen 

very early, even before the completion of uterine involution. iii) 

Additional artificial lightening may retard the time of first pp ovulation. 

Key words: seasonality, reproduction, sheep, ovulation, photoperiod 

804 Concurrent Peste des Petits Ruminants Virus and Pestivirus 

Infection in Stillborn Twin Lambs 

O. Kul 1 , N. Kabakci 1 , A. Ozkul 3 , H. Kalender 2 , H. Atmaca 1 

1 Faculty of Veterinary Medicine, Kirikkale University, Veterinary 

Pathology, Kirikkale, Turkey 

2 Faculty of Veterinary Medicine, Kirikkale University, Animal 

Reproduction, Gynaecology & Obstetrics, Kirikkale, Turkey 

3 Faculty of Veterinary Medicine, Ankara University, Veterinary 

Virology, Ankara, Turkey 

Peste des petits ruminants (PPR) is a contagious viral disease of small 

ruminants that is characterized by pseudomembranous oral lesions, 

bronchointerstitial pneumonia and enteritis. Infection with PPRV is 

generally thought to be transmitted via oral and/or respiratory routes but 

vertical transmission of PPRV has not been reported. In this study, we 

report the pathologic, immunohistochemical and molecular diagnostic 

features of concurrent PPRV and BDV infection in stillborn twin lambs. 

Dead twin lambs were extracted from the birth canal of a 2.5-year-old ewe 

in dystocia in the fifth month of pregnancy and they were necropsied. 

Tissue samples were fixed in 10% buffered formalin and embedded in 

parafin and then sectioned 4-µm in thickness. Unfixed samples of skin, 

spleen, lymph node, lung and brain were submitted for virologic analyses. 

There were 33 ewes in the flock of origin. According to the attending 

veterinarian, clinical findings consistent with PPR, such as oral mucosal 

erosions, coughing, nasal discharge and diarrhea, were observed in this 

flock. Twelve ewes aborted during the lambing season; surviving lambs 

were weak and some had arthrogryposis. The PPRV primers were 

selected to detect the F coding gene of the virus. For pestiviral RT-PCR, 

the selection of primers directed to the p125 protein coding region of the 

BVDV genome was based on the close relationship between ruminant

pestiviruses. Concurrent infection with peste des petits ruminants virus 

(PPRV) and pestivirus was diagnosed in stillborn twin lambs. With the 

flock history, the findings of epidermal syncytial cells and necrotizing 

bronchitis/bronchiolitis prompted testing for PPRV infection and PPRV 

antigen was detected by immunohistochemistry (IHC) in the skin, lungs, 

kidneys, rumen and thymus. Macroscopic anomalies that were typical of 

border disease included scoliosis, brachygnathism, prognathism, 

arthrogryposis, hydranencephaly, cerebellar hypoplasia and hairy fleece; 

pestiviral antigen was detected by IHC in the brain, liver, lungs and 

kidneys. Tissues from both lambs were positive by reverse transcriptasepolymerase 

chain reaction (RT-PCR) for PPRV and pestivirus. To the 

authors’ knowledge, PPR has not been reported previously as a congenital 

infection or in combination with pestiviral infection. 

Key words: Border disease, congenital infection; immunohistochemistry, 

lambs, peste des petits ruminants, pestivirus, polymerase chain reaction 

805 Researches Regarding the Industrial Mating Effect between 

Texel Rams and the Principal Sheep Breeds from Romania 

C. Pascal, M. Ivancia, I. Gilca 

University of Agricultural Sciences and Veterinary Medicine, Faculty 

of Animal Husbandry, Iasi, Romania 

The researches have been made in 2007 year. The goal was obtaining 

of half-breed lambs with very good aptitudes for meat production. For 

this, there have been used male sires from the Texel breeds as paternal 

line and local females (Merinos de Palas, Tsigaie and Turcana races) as 

maternal line. The obtained lambs have been left together with mother 

60 days when the weaning. Since than we have been made the 

homogeneous lots and applied a fattening intensive technology for 85 

days: for starting, 60 days for fattening and 15 days for finishing. At the 

fattening period finish it was established that the half-breed lots have 

made daily average increases and implicitly the superior corporal 

weights with over 20% comparative with those witness, the constant 

differences have been significant statistically talking for p>0,01. After 

finalizing the fattening there were made control slaughter with these 

objectives: determination of weight and structure carcasses, of the 

sacrifice efficiency, of the rapport between basic tissues and muscular 

fibers fineness and muscle surface.After the statistical analyzing of 

slaughter efficiency dates, there was found different that were assured 

statistically, significant between the Texel x Merinos de Palas (DA) and 

Texel x Turcana (DB) lots (p>5%) and very significant between the 

Texel x Merinos de Palas (DA) and Texel x Tsigaie (DC) lots (p>1%) 

and in the other different cases were established insignificant for p 0.05) in the 5 th month, fall to 1.57 

± 0.15 ng/mL (P > 0.05) in 2 nd month of lactation and peak again at 

2,34 ± 0,25 ng/mL in the last month before the next pregnancy. Values 

in BB were higher than in JSB (P > 0.05) in all periods of the year and 

in all reproductive states. 

Conclusions: Leptin values in studied ewes were in the range reported 

for sheep. High values in summer and low in winter reflect 

photoperiod-induced changes in food intake, body weight and 

adiposity, confirmed also in other sheep breeds. Late summer was the 

period of peak breeding activity, explaining high leptin values in 

cycling ewes and confirming the role of leptin in reproductive activity. 

February leptin peak can be explained by high leptin values in late 

pregnancy, observed also in other sheep breeds. They can be attributed 

to high energy intake necessary for the lactation and might originate in 

placenta. Expulsion of placenta after parturition can cause the fall of 

leptin which was observed after the February peak. Annual leptin 

fluctuations are therefore a reflection of environmental conditions, 

modified by reproductive events during the year. 

Key words: ewe, reproduction, leptin 

807 Some Physiological Responses of Sheep under Upper Egypt 

Conditions 

M. El-Fata, A. El-Barody 

Minia University, Anim. Prod. Depart. Fac. of Agric., Minia, Egypt 

Twenty weaned male lambs 2-3 months old with average body weight 

17.75 ± 0.02 kg were used in the present experiment which lasted 180 

days. Animals were randomly divided into five equal groups. The ration 

of the first group (control) was not supplemented with S and I , while those 

of groups 2-5 were supplemented to increase the dietary S and I level up 

to 80 % or 100 % of the maximum tolerable level (MTL) as follows: G2 

(S 80I 80 ), G3 (S 80I 100), G4 ( S 100I 80) and G5 (S 100I 100). The MTL of S was 

0.4 % of feed dry matter, while the MTL of I was 50 PPM of the feed dry 

matter. Blood samples were taken monthly for plasma total protein, 

cholesterol and triiodothryonine hormone determinations. Rectal 

temperature (RT, °C) and respiration rate (RR, r.p.m) were recorded at 1.0 

p.m. during the mid three consecutive days each 30 days. At the end of the 

experiment, two animals from each group were slaughtered. Thyroid 

glands were taken for histological studies. Results revealed that 

differences among animal supplemented groups were significant (P

– Ca, Mg, K, and Ca:P ratio were lower in lactating goat . 

The references ranges for serum biochemical analysis analysis can be 

used for metabolic and nutritional disorders detection in goat 

Key words: mineral metabolism, goat, lactating, Sahara 

810 Metabolic Parameters of Dairy Goats Submitted to Artificial 

Bioclimatic Conditions Similar to the Eastern Amazon Region 

R. Pinho, J. Guimares, J. Ribeiro Filho, L. Martins, E. Castilho, 

M. Borges, F. Santos, E. Da Silva 

Universidade Federal de Viçosa, Veterinary Medicine, Viçosa, Brazil 

This work deals with the adaptability of Alpine and Saanen female goat 

breeds submitted to artificial bioclimatic conditions similar to those of the 

the Eastern Amazon Region, when compared to animals raised under 

normal typical bioclimatic conditions of regions where they demonstrate 

seasonality. The study was conducted from April to July of 2007, 

consisting of an adaptation period of 30 days and an experimental period 

of 60 days. Group 1 (n=4) animals remained in the bioclimatic chamber 

with temperature and air humidity control (8:00-12:00 hours: 30 ˚C; 

12:00-18:00: 36 ˚C; 18:00-8:00: 26 ˚C; with 60% of average humidity; 

and a 12 hour fotoperiod), thus simulating bioclimatic conditions of the 

northern region of Brazil (next to Equator line), whereas group 2 (n=4) 

was kept under influence of natural climatic variations of the season. The 

physiological parameters of respiratory and cardiac frequencies, rectal 

temperature and rumination behavior were measured twice a day, besides 

blood collection twice a week for triiodotironine, tiroxine, cortisol, total 

cholesterol, total proteins and albumine dosages. Water consumption 

values were different, showing that the animals of group 1 consumed 

more than the double (4386.3 mL/day) than those of group 2 (1663.2 

mL/day). The ration consumption demonstrates there a difference both 

between groups and periods (499.0 and 490.8 g/day, for groups 1 and 2, 

respectively). There was no variation in the average values for corporal 

weight and corporal score condition as a function of time, however, 

during the experimental period, there was a difference in the weight of the 

animals between groups (50.5 and 41.5 kg, for groups 1 and 2, 

respectively). A difference was observed for the studied physiological 

parameters between the morning and the afternoon, being the afternoon 

values always higher than the ones in the morning. Cholesterol, albumine 

and total protein concentrations demonstrated no difference between the 

obtained average values during the observation period in function of the 

days. No difference was registered between the obtained values during the 

experimental period in Triiodotironine, tiroxine and cortisol 

concentrations. The results indicated that female goats can be raised under 

bioclimatic conditions, without modifying the related physiological 

standards, making possible the production of embryos over the whole year 

in regions where they demonstrate seasonality. 

Key words: goats, metabolic, bioclimatology 

811 CAE Virus Detection by PCR on Milk Samples 

L. Gregory 1 , M. Lara 2 , M. Hasegawa 1 , E. Meira jr. 1 , H. Rizzo 1 , 

J. Rodrigues 3 , E. Durigon 3 , R. Castro 4 


2 Instituto Biológico, Department of Clinical Science, Faculty of 

Veterinary Medicine, Sao Paulo, Brazil 

3 Universidade de Sao Paulo, ICB, Sao Paulo, Brazil 

4 Universidade Federal de Pernambuco, Virology, Sao Paulo, Brazil 

The caprine arthritis encephalitis is a multisystemic infectious disease 

caused by a lentivirus and may affect animals at all ages and sex. The 

major symptoms are leukoencephalomyelitis, cronical interstitial 

pneumonia and hardening interstitial mastitis. CAE has world wide 

distribution and in Sao Paulo, Brazil it is widely spread among the 

herds. It was described that mammary glands from infected animals 

had a gradual non inflammatory diffuse nodular hardening. It was 

described decreased milk production, high electro-conductivity, high 

levels of chlorides, high cell accounts, low protein levels, low fat levels 

and low solid levels. The CAE virus detection on milk samples by the 

PCR technique is characterized as a method with high specificity and 

sensibility that provides fast results. The technique is able to identify 

proteins or nucleic acids on samples with no living virus. In this study, 

56 milk samples from infected animals were evaluated. 19 samples 

were diagnosed positive by the PCR. The resultant cDNA were used to 

the pol gene amplification. The positive control band was located on 

the 286 pb position, which confirms the adequacy and sensibility using 

PCR for the CAE virus detection on milk samples. 


Key words: CAE, caprine, ruminants, clinic, virology 

812 Caprine Athritis Encephalitis Virus Detection by PCR on 

Tissue Samples 

L. Gregory 1 , M. Lara 2 , M. Hasegawa 1 , E. Meira jr. 1 , H. Rizzo 1 , 

J. Rodrigues 3 , E. Durigon 3 , R. Castro 4 


2 Instituto Biológico, centro de sanidade animal, Sao Paulo, Brazil 

3 Universidade de Sao Paulo, ICB, Sao Paulo, Brazil 

4 Universidade Federal de Pernambuco, Virology, Recife, Brazil 

In 1980 CAE was recognized as a viral disease caused by a lentivirus 

from the Retroviridae family. In Brazil the caprine arthritis encephalitis 

was introduced with irregular acquisition of imported animals.Two 

distinguished clinical manifestations of the disease are characterized by 

the major symptoms described: Leukoencephalomyelitis, that affects 

kids and arthritis, most commonly observed on adult animals. Besides 

these well defined clinical manifestations, it may be observed cronical 

interstitial pneumonia and indurative interstitial mastitis. Some animals 

may be diagnosed negatives when submitted to serological tests due to 

the restricted gene expression, or in the early phases of the disease. On 

these cases PCR tests present itself as a reliable diagnostic technique. 

In this study, 33 tissue samples from brain, lungs and mammary glands 

taken from 11 animals presenting arthritis or mastitis, were submitted 

to PCR. The resultant cDNA was used to the pol gene amplification. 

Two brain samples, one lung sample and one mammary gland sample 

from two animals produced visible positive bands at the agarose gel. 

The positive control band was located on the 286 pb position, which 

confirms the adequacy and sensibility using PCR for the CAE virus 

detection on tissue samples. 

Key words: CAE, caprine, virology, clinic of ruminants 

813 Mercury Poisoning in Goats 

I. Maksymovych 1 , V. Vlizlo 2 , J. Nicpo_ 3 , M. Jankowski 3 

1 Lviv national University of Veterinary medcine and Biotechnolgy, 

Department of Clinical Diagnostics, Lviv, Ukraine 

2 Institute of Animal Biology UAAS, Department of Clinical 

Diagnostics, Lviv, Ukraine 

3 Wroclaw University of Environmental and Life Sciences, 

Department of Internal and Parasitic Diseases with Clinic for 

Horses, Wroclaw, Poland 

Pollution of the environment with heavy metals is registered near the 

industrial centers. Cadmium, lead and mercury are the most toxically for 

organism and do not cause any lesions of different organs and systems. 

Materials and methods: To investigate the clinical symptoms, 

biochemical indices of blood and urine, histological changes in kidneys 

of goats at poisoning with mercury and to study the efficiency of 

treatment measures was the purpose of our research work. 12 healthy 

(control) and 6 mercury poisoned goats of local breed at the age from 9 

months to 3 years were the materials for our investigation. 

Results: Goats poisoning with mercury had caused the depression, 

anorexia, fever, tachycardia, hypotonia of fore-stomach. The 

concentration of urea to 9,7±1,99 mmol/l (P

814 Relationship between Some Blood Parameters and Carcass 

Composition in Native Kid Goats of Boushehr Province (Iran) 

M. Dashtizadeh 1 , MJ. Zamiri 2 , AA. Kamali 1 , AM. Kabirifard 1 

1 Research Center of Agriculture and Natural Resources of Bousheher 

(Iran), Animal Science, Brussels, Iran 

2 Faculty of Agriculture, animal sciences, Shiraz, Iran 

In meat producing chain and improve the genetic potential of strains 

or breeds, there are several stages at which the need for methods of 

measuring composition in live animal arises. One of these methods is 

physiological predictors. The aim of this study was to investigate the 

relationships of some blood parameters and carcass composition in 

native kid goats. Forty-eight 7- to 8-month-old kid goats were 

divided into three equal groups. From each group, 8 goats were feed 

restricted for 45, 60 or 75 days, and 8 goats served as the control 

group. Restricted groups were fed with a maintenance ration, and the 

control goats were fed with a ration supporting 50 g of daily weight 

gain. At the end of feed-restriction period, the restricted goats were 

offered the same ration as the control goats. BUN was positively 

correlated with %CP in DM (r=0.35) and negatively correlated with 

%DM (r= -0.34) and %EE (r= -0.36) and %EE in DM (r= - 

0.39).None of the coefficient between cholesterol and carcass 

composition were significant. Creatinine was negatively correlated 

with %CP (r= -0.34) and %CP in DM (r= -0.36). Also, creatinine 

was positively correlated with %EE (r=0.30) and %EE in DM 

(r=0.37). The results of this experiment showed that creatinine, 

cholesterol and BUN cannot be good predictors of carcass 

characteristics in native kid goats. 

Key words: carcass characteristics, blood parameters, Native Kid 

Goats 

815 Changes in Estrogen and Progesterone Concentration in 

Blood Plasma of Merghoz Goats during Natural Breeding 

Season 

M. Souri 1 , A. Moghaddam 2 , J. Talebi 1 , R. Mirmahmoodi 1 

1 Razi university, Department of Animal Science, Kermanshah, Iran 

2 Razi University/Faculty of Veterinary Medicine, Animal 

Reproduction &AI, Kermanshah, Iran 

Measurement of hormones level associated with reproductive 

function in female animals can aid in the evaluation of their 

reproductive status. The aim of this trail was to determine the profiles 

of changes in progesterone (P4) and Estrogen (E2) in the goat 

peripheral blood. Observation were made on Merghoz does (n=10) in 

the autumn. The animals were housed in individual pens under 

natural day length condition and feed a basal diet. The experimental 

animals received two i.m injections of PGF 2a (PG), 12 days apart. 

Blood samples from the jugular vein were collected once daily 3 days 

before estrus till three days after next estrus. Serum was separated 

and frozen at -20 until analysis. The clinical observations were 

recorded and breeding was allowed after the natural cycle at the third 

estrus. Forty percent and 100% of animals exhibited estrus 72-84 (75 

± 3.0) and 72-78 (72.6 ± 0.06) hours after first and second PG 

injection, respectively. The duration of the estrous cycle was 

recorded as being 20.5 ± 0.37 (18-24) days. A 100% pregnancy rate 

was recorded. The mean length of the gestation period was 145 ± 1.02 

(141-150) days. The initial birth weight of male and female kids were 

2.6 ± 0.08 (2.4-2.8) and 2.2 ± 0.24 (1.5-2.5) Kg, respectively. The P4 

concentration was 2.1 ng / ml on the day -2 days and dropped to the 

basal level (0.46 ng / ml) on the day 0 (Day estrus). The P4 level 

increased steadily from the day 3 (1.6 ng / ml) and attained the 

maximum level of 4.8 ng / ml and 12.5 ng / ml on the days 5 

(metestrus) and 10 (the luteal phase) estrous cycle, respectively. 

From the day 16, the hormone level (10.25 ng / ml) decreased 

gradually and reached to 1.78 ng / ml and basal level of 0.42 ng / ml 

on the days 19 (proestrus) and 21 (Day 0; next estrus), respectively. 

The E2 concentration was 17.12 pg/ml on day -2 (proestrus) and 

reached the maximum level of 22.7 pg/ml on the days 0 (estrus), 1 

and 2. The E2 concentration decreased gradually from day 4 (18.21 

pg/ml) and reached to 17.2 pg/ml on day 7 (metestrus and diestrus). 

The E2 level increased from day 16 (16.29 pg/ml) and attained the 

maximum level of 22.6 pg/ml on day 21 (next estrus). The results 

suggested that the P4 and E2 profile is a useful indicator to assess the 

reproductive status of the Merghoz goats. 

Key words: Merghoz goats, progesterone, estrogen 

816 Comparison of Efficiency of Few Methods of Estrus Induction 

in Out of Season Goats 

M. Dzieciol, R. Kozdrowski, J. Twardon 

Wroclaw University of Environmental and Life Sciences, Veterinary 

Medicine, Wroclaw, Poland 

The objective of this study was to compare the efficiency of few estrus 

induction protocols in anestrus goat, with the use of different ways of 

progestagen administration and different length of treatment. The study 

was conducted in April 2007 in Poland. Sixty five lactating goats having 

had two to three lactations were used. All animals were under the same 

feeding protocol. Does were assigned to five groups (A-E). In the group A 

(n=25) does received intravaginal sponge containing 40 mg of flugestone 

acetate FGA (Chronogest,Intervet) for 12 days, plus intramuscular 

injection of 500 U.I. Pregnant Mare Serum Gonadotrophin (Folligon, 

Intervet), 24 h before sponge removal. The group B gather ten animals 

which received 0,044mg/kg of altrenogest (Regumate, Intervet) per os for 

12 days, and Folligon intramusculary injection (500 U.I.) at the end of 

treatment (day 13). In the group C (n=10) all does received 12,5 mg of 

progesterone i.m. for 6 days. The same protocol (12,5 mg of progesterone 

i.m.) but for 12 days concern does from group D (n=10). All animals from 

group C and D received 500 U.I. PMSG (Folligon) i.m. at the 7 and 13 

days respectively. The goats from group E (control group) did not 

received any treatment. The blood samples for progesterone concentration 

measuring, from 6 goats from every group was collected before and 

during the treatment and after cessation of progestagens administration. 

Following the termination of progestagen treatment, all the does were 

observed for signs of estrus at eight hours intervals for a period of four 

days. Does were mated with fertile males. The four kinds of treatment 

showed significant differences in estrus response. In the group A - 72 %, 

in group C - 60% and in the group D - 100% of animals showed signs of 

estrus. Not any animals from group B showed signs of estrus, but in two 

females from group E estrus signs had been observed. There was no 

significant differences in time of onset of estrus (36,4 h ± 0,5) and the time 

of duration of estrus (40,2 h ± 0,5). At the kidding time 15, 4, 7, and 2 does 

delivered respectively from group A, C, D and E. Considering that 

currently in Poland the vaginal sponge containing progestagens are not 

commercially available, the good results of progesterone i.m. 

administration for estrus stimulation during anestrus are valuable 

information for breeders. 

Key words: goats, estrus induction, progesterone 

817 The Influence of the Hierarchical Structure of Herd, on the 

Answer of Estrus Induction in Anoestrus Goats 

M. Dzieciol, R. Kozdrowski, J. Twardon 

Wroclaw University of Environmental and Life Sciences, Veterinary 


The aim of the study was to find out if the position of the female in the 

herd could play role in the answer on the artificial exogenous hormonal 

stimulation of estrus, and fertility in the out of season goat. The 

experiment was performed in April 2007 in Poland in the commercial 

dairy goat’s herd. Fifty female goats after second or third lactation were 

used in this study. All animals were under the same feeding protocol. 

During the day animals stayed outside, grazed on pastures. Additionally, 

twice a day, during milking time they had access to supplementary food. 

In the first part of investigation the animals were observed and their 

position in herds was qualified. The animals were classified in to the two 

groups trough the direct observation method, according their status in the 

flock, as a-high and low ranked goats (group H, L respectively). The 

estrus induction was performed by using intravaginal sponge containing 

40 mg of flugestone acetate (Chronogest, Intervet) for twelve days with 

intramuscularly injection of 500 I.U. PMSG (Foligon , Intervet) 48 h 

before sponges withdrawal. Also the fertile male was introduced to the 

herd at the time of progestagen cessation. The male was exposed to the 

female in closed pen located on the opposite part of feeding room. We 

observed that the female from group H, which usually used to be as a first 

at the feeding trough exhibited as a first estrus signs. These goats gather 

around the male pen. There was significantly difference in the time of 

approaching male by female and exposing the signs of estrus between 

group H and L. There was no difference in fertility among both groups. 

The results obtained during this study suggest that there is a relation 

between the hierarchical position in herd and the time of estrus signs 

exhibition in goats after estrus induction during anoestrus period. 

Key words: goat, hierarchy, estrus induction 


818 Cerebral Coenurosis in a Goat: Case Report and Literature 

Review 

H. Nourani, K. Pirali Kheirabadi 

Department of Pathobiology, School of Veterinary Medicine, 

Shahrekord University, Shahrekord, Iran 

Cerebral coenurosis is caused by Coenurus cerebralis, the larval stage 

of Taenia multiceps, particularly in goats and sheep. In this case report, 

we describe gross and histopathological characteristics of cerebral 

coenurosis in a goat. An 18-month-old goat with neurologic sings 

including circling and behavioural changes was referred to Department 

of Pathobiology, Faculty of Veterinary Medicine, University of 

Shahrekord. At necropsy, a 4-cm-diameter, fluid-filled and superficial 

cyst with white clusters of scolices was found in the right cerebrum. At 

cut section, in addition to the superficial cyst, a deep compartment was 

seen in the right cerebral hemisphere that caused severe pressure 

atrophy of cerebral gray and white matter. A narrow opening was 

located between these compartments. There were 300 scolices within 

superficial compartment of the cyst and none in the deep compartment. 

Histopatological examination of the affected cerebral hemisphere 

revealed multiple scolices growing on the internal layer of the cyst, 

neuronal degeneration and necrosis, demyelination, hyperemia, 

perivascular cuffing, diffuse microgliosis and astrocytosis. Based on 

the results of this study and other related studies, coenurosis have to be 

considered as differential diagnosis in the cases with neurologic sings. 

Key words: cerebral coenurosis, pathology, goat 

819 Comparison of Concentration of Selenium and Gluthatione 

Peroxidase Activity in Whole Blood of Goats and Their 

Newborn Kids 

L. Misurova, L. Pavlata, A. Pechova, R. Dvorak 

Veterinary and Pharmaceutical University Brno, Clinic for ruminants, 

Brno, Czech Republic 

The aim of this study was to compare concentration of selenium and 

gluthatione peroxidase activity in whole blood of goats and their newborn 

kids. Total of 25 pregnant healthy white shorthaired goats were used in 

experiment. On parturition day, samples of whole heparized blood were 

collected from all the goats. Whole blood samples were also collected 

from their newborn kids before they were given any colostrum. From 

both, goats and kids, samples were taken from v. jugularis. Samples were 

mineralized by microwave digestion technique with HNO 3 and H 2O 2 in 

Milestone Ethos TC appliance. The prepared samples were then tested for 

selenium concentration by the hydrid technique of atomic absorption 

spectrophotometry using the AAS Solaar M6 Unicam spectophotometer. 

The same samples of whole heparinized blood were analyzed for 

gluthatione peroxidase activity photometric by using the Ransel GPx set 

(Randox). Average concentration of selenium in whole blood of goats was 

149.6±45.01 µg/l, average concentration in whole blood of kids it was 

87.91±29.66 µg/l. Average gluthatione peroxidase activity in whole blood 

of goats was 938.46±341.09 µkat/l, in whole blood of kids, it was 658.2- 

339.13 µkat/l. Between concentrations of selenium in whole blood of 

goats and whole blood of their kids were by regression and correlation 

analyses demonstrated statistically very close relationship (p

limb incoordenation. These signs gradually gave place to hind limb 

paresis and finally to ataxia. Severe hyperalgesia and allodynia were 

present on both flanks and hind quarters. There were no seizures, 

hypertermia or change in temperament. Appetite was reduced but 

present. Leukogram revealed a leucocytosis. Lumbar/sacral region X- 

Ray did not show any lesions. Liquid obtained by lumbar puncture was 

negative to microbiology tests. Treatment with antibiotics was 

unsuccessful and the animal was euthanized 6 days after first signs. 

Post-mortem examination revealed cachexia, hind limbs muscular 

atrophy, two well demarcated nodules with 1 cm in diameter in the 

right lung, caseous lymphadenitis and lumbar spinal cord showing 

extensive oedema. Microscopically the lungs showed signs of mycotic 

pneumonia (Cryptococcus sp.) and the lumbar spinal cord showed 

mycotic meningitis with extensive oedema with lymphocyte 

infiltration. A diagnosis of cryptococcal meningitis was possible due to 

microscopic lesions. Lymphadenitis was caused by C. pseudotuberculosis. 

Cryptococcus neoformans seems to have a predilection 

for the central nervous system in humans. Cryptococcal meningitis has 

been described in imunnocompromised human patients and cats but no 

report of goat lumbar-meningitis due to Cryptococcus infection has 

been published.Pigeons are thought to be the main carriers of 

Cryptococcus neoformans. Pigeons had access to both the stable and 

the pastures used by the buck but no other animal in the herd has shown 

any neurological signs. A few goats in the herd have shown respiratory 

disease signs for which a mycotic etiology has not been confirmed. 

Key words: goat, mycotic meningitis, allodynia, neurologic signs 

823 Periparturition Dynamics of Serum Protein Levels in Goats 

L. Janku, L. Misurova, J. Filipek, A. Pechova, R. Dvorak, L. Pavlata 

University of Veterinary and Pharmaceutical Sciences, Faculty of 

Veterinary Medicine, Clinic of Ruminants, Brno, Czech Republic 

The aim of the study was to compare levels of individual protein fractions 

(PF) in blood serum of goats determined by electrophoresis (ELFO) in the 

periparturition period (p.p.). Eight clinically healthy white shorthaired 

goats were examined. Blood samples of these goats were repeatedly taken 

from v. jugularis 3 weeks (wk) before the parturition (p.), on the 

parturition day (p.d.) and on day 28 after the parturition (a.p.). Each PF 

was identified using Hydragel Protein K20 kit. Agarose gel ELFO divided 

PF to albumins (A), alfa1, alfa2, beta1, beta2 and gama-globulins 

(gamaG). Separated PF zones were stained and evaluated by 

densitometry. Percentage shares of individual PF were converted from 

total protein (TP) values to g/l values. The results indicated that serum PF 

in goats in the p.p. show significant dynamics of change. No significant 

differences between samples taken 3 wk before the p. and on the p.d. 

(except for gamaG that decreased from 9.5±2.6 to 8.1±1.7 g/l) were 

determined (the values obtained 3 weeks before the p. vs. the p.d. values 

were as follows (in g/l): TP 60.8±3.8 vs. 60.6±4.3, A 36.7±1.7 vs. 

37.4±2.9, alfa1 3.5±0.5 vs. 3.5±0.6, alfa2 3.1±0.5 vs. 3.4±0.5, beta1 

5.5±0.3 vs. 5.6±0.5, beta2 2.5±0.3 vs. 2.6±0.2). However, most of the 

values from day 28 a.p. were significantly higher (TP 71.3±2.4, A 

42.7±1.0, alfa2 4.1±0.8, beta2 6.3±0.9, gamaG 9.9±1.6 g/l). The only 

fractions that did not changed significantly during the period were alfa1 

and beta2 (day 28 values were 3.3±0.6 or 2.5±0.6 g/l respectively). 

Interpretation of PF results must consider the stage of reproduction and 

lactation, as these significantly affect most of the monitored parameters 

(TP, A, alfa2, beta1, gamaG) also in healthy animals. Three wk before the 

p., many values obtained from healthy animals were substantially lower 

than those measured during lactation period. Lower values identified 

before the p. should be associated above all with the production of 

colostrum and other immunological changes that happen in the p.p. As we 

monitored healthy animals only, we believe that not even increased values 

of fractions involving acute-phase proteins can be associated with any 

disease. 

The work was supported by the Research Project MSM6215712403. 

Key words: protein fractions, blood, globulins, electrophoresis 

824 Supplementing Goats in Semi-arid Conditions with Maize 

during the Last 12 Days of Pregnancy Improves the Ability of 

New-born Kids to Recognize Their Mother 

H. Hernandez 1 , S. Ramírez 1 , J. Flores 1 , P. Poindron 2 , A. Terrazas 3 , 

J. Delgadillo 1 

1 CIRCA, UAAAN, Torreón, Coahuila, Mexico 

2 UMR 6175, INRA, Tours, France, France 

3 FESC, UNAM, Mexico, Mexico 

Objectives of the study: We investigated if a supplementation with 

maize during the last 12 days of pregnancy could improve the ability of 

newborn kids to reunite with their mother and recognize her in goats 

grazing natural pasture under semiarid conditions. 

Materials and Methods: A group of 21 control goats (C group) were 

maintained under traditional extensive management, in which the 

animals were taken out daily to graze the available natural vegetation 

until parturition. Another group of 31 supplemented goats (S group) 

were managed in the same conditions as the controls, but received in 

addition 0.6 kg of flaked maize/mother/day during the last 12 days of 

pregnancy. At 8 hours postpartum one kid from each mother underwent 

a 5 min two-choice test between their own mother and an alien goat that 

had given birth at about the same time in a 4x4 m enclosure. Thus, 21 

C kids (4 singles, 11 twins and 6 triplets) and 31 S kids (7 singles, 15 

twins and 9 triplets) were tested. 

Results: Fewer C kids than S kids reached a mother during the test 

(9/21versus 23/31; P0.05). In contrast, in the S group 

all these variables were significantly higher toward the own mother than 

toward the alien mother (time near a mother: 135.8 ± 20.0 s vs 18.7 ±12.0 

s; time looking: 117.7 ± 18.0 s vs 17.0 ± 4.1 s; number of visits: 1.4 ± 0.2 

vs 0.3 ± 0.2; P

concentrations of the goat milk, including colostrum, suggesting the 

viability of this thermal treatment, used as alternative to prevent 

arthritis encephalitis syndrome (CAE) in newborn kids. 

Key words: goat, milk, colostrum 

826 Concentrations of Progesterone, Estradiol-17ß and Insulin 

like Growth Factor-1 in Baladi Goats after Recombinant 

Bovine Somatotropin (rbST) Administration 

K. El-Battawy 

National Research Centre, Animals Reproduction, Cairo, Egypt 

This investigation was carried out to study the impact of rbST 

administration on the levels of progesterone, estradiol-17ß and insulin 

like growth factor-1 in Baladi goats. Eleven Baladi goats aged 2-3 

years were used in this investigation after being assigned randomly into 

three groups. First group (rbST+LH group, n=4), each goat was 

injected SC with 10mg rbST in 2ml buffer saline daily, starting from 

day 5 of synchronized estrous cycle till day 12 of cycle then S/C 

pregnyl (LH) was injected at a level of 275 I.U. per animal while each 

animal in the second group (rbST group, n=4) was administered only 

rbST as previously mentioned. A third group (n= 3) served as control. 

Goats were exposed to mature, fertile bucks daily and blood samples 

were collected daily. Results indicated that the progesterone 

concentrations and estradiol-17ß. 

In conclusion, this study confirmed that the administration of rbST 

increased the levels of progesterone, estradiol-17ß and IGF-I 

827 Stimulation of Oestrous Activity of Female Goats Exposed to 

Long Days Treated Sexually Active Males in Different 

Months of Seasonal Anoestrous 

J. Flores, E. Carrillo, F. Veliz, H. Hernanadez, J. Delgadillo 

Centro de Investigación en Reproducción Caprina, Universidad 

Autónoma Agraria Antonio Narro, Departamento de Ciencias Médico 

Veterinarias, Torreón, Coahuila, Mexico 

Objective: This study was carried out to determine the response of 

female goats to male effect in different months of seasonal anoestrous 

using sexually male goats treated with continuous long days and 

untreated males. 

Material and Methods: In three months of seasonal anoestrous (April, 

May and June), three groups of anovulatory does (a group each month) 

were exposed during 15 days to males rendered sexually active with a 

treatment of continuous long days (16 h light/day), while other three 

groups of does (a group each month) were exposed to untreated males 

that only perceived the natural variations of photoperiod (26°N). The 

oestrus was detected twice daily during 15 days. The sexual behaviour 

of males (nudging, mount intention and complete mounts) was 

recorded during 2 h in the first 5 days after male introduction. 

Results: In April, the proportion of females showed oestrous behaviour 

was higher in does exposed to treated (18/19) than untreated males (0/20; 

P0.05) between does stimulated with 

treated (20/20 and 18/20, during May and June, respectively) and untreated 

males (19/20 and 18/19, during May and June, respectively). The sexual 

behaviour of the males treated with continuous long days was greater than 

the control males (P0.05). 

Conclusions: These results show that oestrous activity of female goats 

can be stimulated during April only when sexually active males are 

used. In contrast, during May or June, the response of females is similar 

when exposed to treated or untreated males. The lack of difference is 

likely to be due to the onset of the natural sexual activity of untreated 

males, which start in May. 

Key word: goats, sexually male goats, anestrus, estrous 

828 Exposure to Artificial Long Days During Winter Increases 

Milk Production in Saanen Goats from Subtropics 

F. Véliz 1 , J. Flores 1 , J. Elizundia 2 , H. Hernandez 1 

1 CIRCA, UAAAN, Torreón, Coahuila, Mexico 

2 Dirección Particular: Angel Camino 161, Col. Valle del Nazas, 

Gómez Palacio, Durango, Mexico 

Objectives of the study: The aims of the present study was to 

investigate 1) if artificial long days (16 h of light/day) during winter 


may increase the milk production in Saanen goats from subtropical 

region and 2) if the milk production is maintained after remove the 

photoperiodic treatment. 

Materials and Methods: To this end, we used two balanced groups of 

Saanen goats with regard to date of parturition, initial milk production, 

parity, live weight and condition score. Starting on December 16 th (day 8 

± 0.6 postpartum; mean ± sem) one group of goats (n=19) was submitted 

daily during first 70 days of lactation to artificial long days (16 h light 

/day; Treated group), followed by the natural short days. Simultaneously, 

another group of goats (n=19) were maintained under natural variations of 

photoperiod (nearly 10 h and 19 min of natural light/day during winter 

solstice; Control group) during first 134 days of lactation. In both groups 

the weaning of the kids was performed at 30 days postpartum. All goats 

were maintained under intensive conditions and were fed alfalfa hay (with 

18% of CP) and commercial concentrate (14% CP). Each two weeks, in 

all animals milk production in a 24 h-period were assessed through two 

machine milking 12-h apart up to 134 days postpartum. 

Results: Before the weaning of the kids, milk production did not differ 

significantly between the two groups (3.1 ± 0.1 kg in both groups; 

P>0.05). In contrast, at days 42, 56 and 70 post-exposition, goats from 

long days treated group yielded in average 3.0 kg of milk/day, and 

goats from control group only 2.5 kg of milk/day (P

2 Veterinary Medicine Faculty, Dept. Physiological, Biochemical and 

Cellular Sciences, Sassari, Italy 

Objectives of study: Energy metabolism is a key factor supporting cell 

functions and male gamete require exceptionally more ATP than other 

cell types in order to sustain its high queue activity. The aim of this 

study was to evaluate the relationhips between ATP concentration, 

viability and motility in fresh and cryopreserved-thawed spermatozoa 

during the breeding season. 

Materials and Methods: During the breeding season, semen was 

collected by artificial vagina from three bucks originating from native 

stocks in Ogliastra area of Sardinia. Semen was cryopreserved in 4% 

glycerol, cooled to 4 °C, frozen in 0.25 ml pellets on dry ice and then 

plunged into liquid nitrogen. Fresh and frozen/thawed semen samples 

from the three males were evaluated for viability by PI fluorescent 

staining and for progressive motility by Sperm Class Analyzer (v. 

3.2.0. Microptics S.L., Barcelona, Spain), while ATP levels were 

measured spectrophotometrically at 340 nm using NADH-linked 

enzyme-coupled assays (H.U. Bergmeyer, Principles of enzymatic 

analysis, Academic Press, New York ,1974) modified for being 

adapted to the our system. Data were analysed using Statgraphics 

Centurion XV (Statpoint, inc). 

Results: Our data show that both ATP, progressive motility and 

viability do not differ between subjects nor between ejaculates in both 

fresh and frozen semen (Anova; P

was significant and showed that breeding in Miankale buffaloes which 

are mixed breed of water buffaloes and are living in wild condition can 

be done successfully. 

Key words: ovsynch cider, buffaloes, Miankale peninsula, Iran 

834 Isolation of Lactobacillus from the Vagina of Buffalo-cows 

and its in vitro Probiotic Activity 

S. Abd El-Moez1 1 , W. Ahmed 1 , J. El Jakee 2 , F. El Seedy 3 

1 

National Research Center, Animal Reproduction &AI, Giza, Egypt 

2 

Faculty of Veterinary Medicine, Microbiology, Giza, Egypt 

3 

Faculty of Veterinary, Beni Sueif University, Microbiology, Beni 

Sueif, Egypt 

Lactobacilli are among the predominant microorganisms in the vaginal 

tract of some homoeothermic animals. The current investigation was 

designed to throw line on Lactobacillus in the vagina of buffaloescows. 

Vaginal and blood samples were collected from animals reared at 

Lower Egypt for a period of 2 years. Vaginal swabs were collected 

from buffalo-cows either showed normal ovarian activity or suffering 

from ovarian inactivity. Swabs were collected from each animal in 

either Tryptic soy broth for the routine bacteriological examination or 

De Man, Rogaso, Sharpe (MRS) for isolation of Lactobacillus. In vitro 

sensitivity and probiotic tests against the most predominant isolates 

isolated from buffalo cows (Y. enterocolitic, C. diversus, E. coli, 

Micrococcus spp., E. Faecalis, S. aureus and Bacillus spp. were carried 

out). Results showed that Lactobacillus was isolated from the vagina of 

normal cyclic buffalo-cows with an high incidence (90.91%) as 

compared to in animals suffering from ovarian inactivity (81.82%). 

Vaginal pH and the recovered isolates were recorded in relation to 

ovarian activity. Ciprofloxacin and ofloxacin are the most effective 

antibiotics followed by tobramycin, gentamicin and oxytetracycline. 

The resistant rate of isolates to some antibiotics was recorded. In vitro 

use of Lactobacillus isolated from the genital tract of normal buffalocows 

as probiotic against the most predominant isolates from buffalo 

cows suffering from ovarian inactivity revealed that L. acidophilus is 

the most effective strain followed by L. casei rhamnosus. It could be 

concluded that Lactobacillus is one of the normal bacterial floras of the 

vagina of buffalo-cows; L. acidophilus and L. casei rhamnosus were 

the most predominant isolates and showed high probiotic effect against 

the most predominant bacteria isolated from cases suffering from 

ovarian inactivity. 

Key words: buffaloes, vagina-Lactobacillus, antibiotics, probiotics 

835 Efficacy of Toltrazuril (Baycox Bovis) and Diclazuril 

(Vecoxan) in the Control of Subclinical Coccidiosis in Buffalo 

Calves 

M. Diaferia 1 , F. Veronesi 1 , G. Severi 1 , D. Lulla 2 , L. Nisoli 2 , 

D. Piergili Fioretti 1 

1 Faculty of Veterinary, Department of Parasitology, Perugia, Italy 

2 Bayer Health Care, Milan, Italy 

Coccidiosis in buffalo species is a widespread protozoa infection 

caused by several species belonging to genus Eimeria and localized in 

the intestine. Higher values of prevalence and diarrhoeic 

manifestations are more frequently observed in young animals; 

however, subclinical coccidiosis is responsible for serious economic 

losses for reduction of growth rates and feed conversion in young 

animals. The objective of this field efficacy study was to demonstrate 

the effect of a treatment with Toltrazuril (Baycox Bovis ® ) and 

Diclazuril (Vecoxan ® ) on buffalo calves naturally exposed to Eimeria 

infection, in terms of lack of faecal oocyst excretion and body weight 

gain. Two farms in Central Italy were selected based on previous 

history of coccidiosis outbreak by E. zuernii and E. bovis and on the 

presence of subclinical infections. A total of 135 buffalo calves ranged 

from 4 to 15 weeks of age and from 39 to 80 kg and positive for 

coccidiosis, were selected. The animals were divided in 3 distinct 

groups (T, D and C) of 30 (farm A) and 15 subjects (farm B), each 

based on population size of the farms. On day 0 the T group was treated 

with Toltrazuril (Baycox ® Bovis) at dose of 15 mg/kg body weight; the 

D group was treated with Diclazuril (Vecoxan ® ) at dose of 1 mg/kg 

body weight and the C group was kept as untreated control. The 

animals were examined for the presence of Eimeria species by qualiquantitative 

coprological tests at 0,2,4,7,49,51,53,56 days posttreatment. 

No side effects were observed in the medicated group during 

the trial. On day +7, Toltrazuril showed a mean efficacy value of 98. 


85%. High values of efficacy were observed till 42 days after treatment; 

on day 56 the mean value was of 65.37%. On day +7 Diclazuril showed 

a mean efficacy value similar but lower than Toltrazuril (97.28% versus 

98.85%); values > 50% were observed only till 23 days after the 

treatment. Statistical significant difference was observed between the 

animal of the T and D groups and the control group (p

thermography. Two groups of 14 buffaloes were milked at 42 kPa, high 

vacuum (HV) and, after 3 weeks of adaptation to progressively lower 

vacuum levels, at 36 kPa (LV), using the same pulsation parameters (60 

cycles/min and 65:35 ratio). Milking was carried-out at intervals of 9 

hours (daytime) and 15 hours (over-night). Tests were performed in a 

28+28 units parallel parlour with low line milking system equipped with 

light weight clusters (1.78 kg), automatic cluster removers and 

electronic herd management system. Thermographic images (Flir 

System, ThermaCam P25) of posterior udder area (PUA) and teats 

(base-teat - BT; middle-teat - MT and tip-teat - TT) were taken premilking 

(PM), during milking (M), immediately after milking (IAM) 

and up to 5 minutes after milking (AM+). Temperatures were recovered 

by processing the thermoimages in ThermaCam Researcher Basic 2.8 

Software. The results of vacuum level effects on teat tissue temperature 

show evident differences among BT and TT, and among MT and TT in 

all stages. Moreover interesting is the temperature difference between 

LV and HV levels at PM and IAM: relevant gap is showed in HV level 

(2.67 °C) compared to LV (1.79 °C), underlining a greater blood flow 

arrangement and smaller distress of teat. Though being an introductive 

work, this data set suggests an important role of thermography to 

monitor the effects of milking procedures on udder and teat. 

838 Serum Levels of Gamma Glutamyl Transferase (GGT) and 

Aspartate Aminotransferase (AST) in Murrah Buffaloes 

V. Gomes 1 , K. Madureira 1 , K. Leonel 1 , M. Blagitz 2 , S. Kitamura 3 , 

A. Gomes 4 , F. Benesi 2 

1 Faculdades Anhanguera, Medicina Veterinária, Leme, Brazil 


3 

Universidade Anhembi Morumbi, Medicina Veterinária, Sao Paulo, 

Brazil 

4 

Universidade Estadual do Norte do Parana, Medicina Veterinária, 


Although buffaloes are affected by several hepatic diseases, diagnosis 

of these conditions only based on liver examination is difficult. Due to 

this fact, the use of complementary exams, such as serum activity of 

liver enzymes GGT and AST is very important. However, there are no 

reference values determined for buffaloes that would aid the 

interpretation of the results. Therefore, the objective of the present 

study was to determine reference values and to assess the influence of 

age on serum activity of GGT and AST in Murrah buffaloes bred in Sao 

Paulo, Brazil. Serum samples collected from 125 clinically healthy 

buffaloes and were placed into 4 experimental groups, according to the 

age: 0 to 3 months of age (Group 1); 4 to 6 months of age (Group 2); 7 

to 12 months of age (Group 3); and over 1 year of age (Group 4). GGT 

and AST serum activities were evaluated using commercial kits in an 

automatic biochemical analyzer at 405nm. GGT serum activity was 

equal to 35.9; 19.6; 25.1; and 33.3 UI/L, respectively, in animals of 

Groups 1, 2, 3 and 4. Activity was significantly greater in the first 

months of life. AST serum activity, on the other hand, was significantly 

greater in the first months of life (92.3 UI/L), increasing to 128, 177 and 

143 UI/L as animals grew older in groups 2, 3 and 4, respectively. 

Therefore, it was concluded that age is a physiological factor that 

causes variations in the serum activity of liver enzymes GGT and AST. 

Besides, reference values were determined for the enzymes, which is 

an excellent aid to the clinical diagnosis of liver diseases in domestic 

animals. 

Key words: Murrah buffaloes, blood, gamma glutamyl transferase, 

aspartate aminotransferase 

839 Total Protein, Albumin and Globulin Serum Concentrations 

in Murrah Buffaloes. Reference Values and Influence of Age 

V. Gomes 1 , K. Madureira 1 , K. Leonel 1 , M. Silva 1 , M. Blagitz 2 , 

A. Gomes 3 , S. Kitamura 4 , F. Benesi 2 

1 

Faculdades Anhanguera, Medicina Veterinária, Leme, Brazil 

2 

Universidade de Sao Paulo, Clinica Medica, Sao Paulo, Brazil 

3 

Universidade Estadual do Norte do Parana, Medicina Veterinária, 


4 

Universidade Anhembi Morumbi, Medicina Veterinária, Sao Paulo, 

Brazil 

Proteins are indispensable to life due to their biological significance 

and to the various functions they perform in the organism. Because of 

this, the evaluation of protein serum concentration is an important tool 

in clinical diagnosis. Although it is important to know the physiological 

biochemical profile for an adequate interpretation of results when 

serum proteins are evaluated, there are few studies on the subject 

involving buffaloes. In the present study, serum concentrations of total 

protein, albumin and globulin were determined in 125 healthy Murrah 

buffaloes. Animals were divided into 4 experimental groups, according 

to the age: 0 to 3 months of age (Group 1); 4 to 6 months of age (Group 

2); 7 to 12 months of age (Group 3); and over 1 year of age (Group 4). 

Serum concentration of total proteins was carried out by means of the 

biuret method, whereas serum albumin was determined by means of 

bromocresol green. Globulin concentration was determined by 

subtracting individual values obtained for total protein and albumin. 

Values were expressed in g/dL. Mean concentration of serum protein 

was greater in animals of group 1 (8.2 g/dl) and 4 (7.7 g/dL), and lower 

in animals of group 2 (7.0 g/dL) and 3 (6.9 g/dL). Albumin 

concentration was equal to 2.4; 2.2; 2.1 and 2.5g/dL in animals of 

groups 1, 2, 3 and 4, respectively. Concentrations were greater in 

groups 1 and 4. Globulin concentration was equal to 5.8; 4.7; 4.7; and 

5.1, in groups 1, to 4, respectively. Therefore, age seemed to influence 

the biochemical characteristics studied, with greater total protein, 

albumin and globulin concentrations mainly observed in group 1. 

Key words: Murrah buffaloes, total protein, albumin and globulin 

840 A Survey on some Bacteriological and Pathological Aspects of 

Buffalo Liver Abscesses 

A. Ghadrdan-Mashhadi 1 , M. Jamshidian 2 , K. Gheysar-Beigi 3 

1 

Faculty of Veterinary Medicine, Shahid Chamran University, 

Department of Clinical Sciences, Ahvaz, Iran 

2 


Department of Pathobiology, Ahvaz, Iran 

3 

Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, 

Iran 

Local suppurative infections of the liver do not cause clinical signs of 

hepatic dysfunction unless they are particularly massive or extensively 

metastatic, however it sometimes causes to weight loss. Also, they do 

cause significant losses in feedlot and grain-fed animals because of 

rejection of the affected liver at the abattoir. This study was carried out 

to find the occurrence of liver abscesses in buffaloes slaughtered in 

Ahvaz in 2003. The animals (n: 503) were selected from two sexes and 

divided to three age groups (group A: less than 2.5 year, group B: 2.5- 

5 years, and group C: more than 5 years). If the abscesses were present, 

the location and sizes would be recorded and then sampling for 

bacterial culture was done. Results showed that 5 buffaloes (0.99%) 

had liver abscesses. According to presence of abscesses, there was no 

significant difference between two sexes and three age groups. Most of 

the abscesses were found in right lobes (40%) and in diaphragmatic 

surface (60%) of livers. The following bacteria were isolated: A. 

pyogenes (3 cases) and S. epidermidis (1 case). In one case the abscess 

was sterile. 

841 The Anatomy of Buffals Cardiac Veins 

HA. Karimi 1 , GH. Ardalani 2 , AH. Zadeh Kokia 

1 Faculty of Veterinary Medicine, University of Tabriz, Basic 


2 Faculty of Veterinary Medicine, Islamic Azad University of 

Shahrekord, Basic Sciences, Urmia, Iran 

In this research anatomical structure of coronary vein and type of low 

oxygen blood drainage into right atrium was studied in Gharbi 

Azarbayjan buffalo heart. 20 healthy buffaloes’ hearts and healthy cows’ 

hearts are collected from Tabriz industrial slaughter house. Buffaloes’ 

hearts are used for studying coronary veins and cows’ hearts are used for 

comparating to buffaloes’ hearts. Anatomical structure of buffalo 

cardiatic veins (coronary veins) are detected by injection contraste meida 

such as Barium Sulphat and magnulin and radiograph was prepared. Also 

the hearts were injected by coloured latex, rodopass and ejllatine and this 

injected buffaloes’ hearts are dissected and compared to cows’ hearts. 

Obtained results of the survey are showed that buffaloes cardiatic blood is 

drining into right atrium by two veins, great cardiatic veins and middle 

cardiatic veins. Great cardiatic vein lies on left longitudinal groove. It was 

parallel to left longitudinal branch of left coronary artery. Great cardiatic 

vein arises a circumflex branch that lies on coronary groove between left 

atrium and left ventricle. Great cardiatic vein receives blood from left 

atrium and left ventricle and it drains into coronary sinus. Obtained results 

of coronary sinus biometry are showed that coronary sinus has about 4/5 


cm long and 1/48 cm diameter. Middle cardiatic vein goes towards 

coronary sinus and it drains into it. This vein and interventricular 

subsinousal artery together lie on right longitudinal groove. Another large 

vein is observed in buffaloes’ heart. It is called ventricular margine vein. 

Results of this survey are established that buffaloes’ heart is different from 

other ruminants’ heart. Ruminants’ heart is monocoronary type but 

buffaloes’ heart is bicoronary. 

Key words: anatomy, buffalo, heart, vein 

842 Plasma Concentrations of Testosterone and Androsteneioda 

in Male Buffaloes (Bubalus Bubalis) Originated from 

Tropical Regions 

J. Siqueira, E. Oba, R. Bittencourt, L. Leal 

FMVZ-UNESP, 1Department of Animal Reproduction and Veterinary 

Radiology, Botucatu-SP, Brazil 

Despite of the important functions that the hormones play in physiology, 

it is observed that few studies about reproductive endocrinology are 

related to the male and even less to bufaline species (Ohashi, Rev Bras 

Reprod An, 299:31, 2007). Thus, the objective of this study was to 

determine the plasma levels of testosterone and androstenedione in 35 

crossbred buffaloes (Murrah x Mediterranean), raised extensively in 

tropical region. The animals had 27.0 ± 4.08 months of age and 190.5 ± 

31.79 kg of carcass weight on average. The blood of animals was 

collected during the slaughter, transported under refrigeration to the 

laboratory (FMVZ/UNESP/Botucatu-Brazil) and centrifuged (2,500 g) 

for 15 minutes. The plasma was stored at -70 °C. The hormonal 

determinations were performed by the method of radioimmunoassay 

(RIA). The SAS statistical program was used to evaluate mean, standard 

deviation and Pearson’s correlation between the analyzed characteristics. 

The mean values and standard deviation of testosterone and 

androstenedione concentrations were respectively 124.98 ± 106.39 pg/ml 

and 221.14 - 272.10 ng/ml, results similar were found by Ohashi (Buffalo 

Journal, 12:313 -320, 1996) in crossbred buffaloes with 18 to 24 months. 

The testosterone concentration presented a positive correlation with body 

weight and androstenedione concentration (p

2 Institute for Veterinary Pathology, Centre for Fish and Wildlife 

Health, Vetsuisse Faculty, University of Bern, Berne, Switzerland 

3 Chembio Diagnostic Systems, Inc., 3661 Horseblock Road, Medford, 

New York 11763, United States 

Objectives of study: The goal of the study was to describe the clinical 

findings, clinicopathologic abnormalities, serological tests, diagnostic 

imaging findings and necropsy results in South American Camelids 

(SAC) infected with Mycobacterium microti, a member of the M. 

tuberculosis complex. 

Materials and Methods: The records of 10 animals (9 llamas and 1 

alpaca; aged 4 to 18 years) from 4 different herds with a history of wasting 

and weakness admitted to the Clinic for Ruminants and / or the Institute 

for Animal Pathology within a timeframe of 6 years were reviewed. 

Results: Clinical signs were limited to weight loss, recumbency and 

anorexia in late stages of the disease. The single comparative 

intradermal tuberculin test with bovine protein purified derivate (PPD) 

and avian PPD was negative in all animals. No consistent hematologic 

abnormalities were identified. Animals that were tested serologically 

positive (multiantigen print immunoassay, lateral-flow-based rapid 

test) were examined in detail with abdominal and thoracal ultrasound 

and thorax radiology. Abnormal findings such as enlarged mediastinal, 

mesenterial and /or hepatic lymph nodes were seen by ultrasound only 

in advanced cases. The infection was confirmed at necropsy in all 

animals by bacteriological culture and / or spoligotyping. 

Conclusions: An infection caused by M. microti should be considered 

a differential diagnosis in chronic debilitating diseases with or without 

respiratory signs in SAC. However, the clinical diagnosis remains 

challenging particularly in the early stages of the infection. 

Key words: lama, alpaca, tuberculosis, Mycobacterium microti 

140 Point Prevalence of Bacterial and Protozoal Intestinal 

Pathogens in Suckling Camel Calves in Northern Kenya 

I. Gluecks 1 , M. Younan 2 , S. Bornstein 3 , C. Ewers 4 , W. Mueller 5 

1 Veterinaires Sans Frontieres Suisse, Kenya Programs, Nairobi, 

Kenya, Kenya Coast Republic 

2 Veterinaires sans Frontieres Germany, Somalia Program, Nairobi, 


3 National Veterinary University of Agricultural Science, Department 

of Parasitology, Uppsala, Sweden, Sweden 

4 Veterinary Faculty, Free University Berlin, Institute for 

Microbiology and Epizootics, Berlin, Germany 

5 Veterinary Faculty, Free University Berlin, Institute for Animal and 

Environmental Hygiene, Berlin, Germany 

Objective of the study: This study was conducted from 2002 to 2004 in 

Northern Kenya in order to investigate the prevalence of bacterial and 

protozoal intestinal pathogens in camel calves up to 12 weeks of age. 

Material and Methods: A point prevalence study was conducted to 

describe the existing intestinal pathogens according to age groups, 

health status and to compare their occurrence between two 

management systems. From each camel calf between birth and 12 

weeks of age belonging either to ranch (n=157) or pastoralists (n=72) 

herds a faecal sample and rectal swab was taken for bacteriological and 

parasitological analysis. 

Results: of the 229 individual camel calves sampled in both management 

systems, 67.7% were healthy, 23.1% exhibiting diarrhoea, 6.6% 

convalescent and 2.2% dead. A higher percentage of camel calves 

suffering from diarrhoea were found in pastoralist herds (31.9%) as 

compared to ranch herds (19.2%). There was a peak of camel calves 

suffering from diarrhoea within the second and third week of age. 

Isospora sp. and Strongyloides sp. were excreted in 6.6% while only in 

4.6% Strongyle sp. eggs were excreted. Isospora sp. excretion was more 

prevalent in pastoralist herds (12.9%) as compared to ranch herds (3.7%). 

Excretion of Isospora sp. was most prevalent from the second till seventh 

week of age, no shedding was diagnosed at an older age. Klebsiella 

pneumoniae was isolated in 26.9% (n=119), Salmonella sp. in 19.1% 

(n=226), and E. coli in 97.5% (n=200) of calves sampled. The point 

prevalences of K. pneumoniae and Salmonella sp. were particularly high 

in the first three weeks of age. Sequence analysis of the SSU rRNA gene 

and ITS 1 confirmed that the Isospora sp. isolates from this study 

belonged to the species Isospora orlovi. Out of 32 K. pneumoniae positive 

camel calves 18 different capsular antigens types were identified. 

Salmonella bovismorbificans was the most common serotype (32.6%), 

followed by S. butantan (21.5%), S. typhimurium (11.1%), S. kiambu 

(9.0%) and S. muenchen (7.6%). In 78 of the E. coli isolates virulence- 

associated genes were detected: eae (13.7%), astA (10.6%), hlyEHEC 

(4.3%) and stx (2.0%). None was positive for elt Ia/Ib and est Ia/Ib. 

Conclusion: The importance of the different pathogens in the health 

status of the camel calf and the differences occurring between the two 

management systems are discussed. 

Key words: camels, calves, diarrhoea, pastoralism, Kenya 

141 Mastitis in Camels in Somalia and North Kenya 

I. Gluecks 

Veterinaires sans Frontieres Germany, Somalia Program, Nairobi, 


In semiarid regions of the Greater Horn of Africa, in particular North 

Kenya and Somalia, camels are the most important dairy animal for 

pastoralists and play an important role in food security of nomadic 

households. In semiarid environments camels produce 2.5 times more 

milk than cows and are less dependent on water sources. Information 

on prevalence and importance of mastitis in pastoralist camels is very 

limited and control concepts to limit the impact of mastitis on milk 

production of camels do not exist. Several field studies were 

undertaken in camels in Somalia and North Kenya to obtain baseline 

data on mastitis prevalence and on applicable mastitis management 

concepts. Camel herds were visited once during the morning milking. 

Lactating camels were examined clinically, tested by California 

Mastitis Test and sampled individually. Milk was taken in an 

uninterrupted cold-chain to an ISO-certified private laboratory in 

Nairobi (Analabs Ltd.) where bacteriological cultures for the two most 

important infectious mastitis pathogens (Streptococcus agalactiae, 

Staphylococcus aureus) were carried out. Due to security constraints in 

Somalia access to camel herds is very limited. In order to gain an 

insight into the prevalence of infectious mastitis pathogens raw camel 

milk sold on local markets was also sampled and 

analysed. Streptococcus agalactiae was found in 70% of camel milk 

container sampled at urban Somali milk markets and was present in 5 

of 10 examined lactating camel herds. 42% of udders tested in two 

herds were CMT-positive and 34% of these were infected by 

Staphylococcus aureus. In North Kenya loss of one or more quarter 

was seen in 8% of 207 camels examined individually and the traditional 

practice of teat tying was identified as one contributing factor. Camel 

owners mostly focus on the few clinical mastitis cases but are unaware 

of widespread subclinical intramammary udder infections. Reduction 

of the economically most important infectious mastitis pathogens 

(Streptococcus agalactiae and Staphylococcus aureus) in camel herds 

is desirable but no treatment or control concepts have been tested in 

lactating camels so far. Unlike in cattle a healthy nasopharyngeal 

carrier state for Streptococcus agalactiae exists in camels, which raises 

doubts about success of eradication attempts. Constraints to mastitis 

treatment and management options to reduce the prevalence of 

subclinical mastitis in pastoralist camels are discussed. 

Key words: mastitis, camels, Somalia, Kenya 

142 Bilateral Surgical Fracture Repair in an Alpaca 

J. Declercq 1 , G. Vertenten 1 , L. Devisscher 1 , V. Barberet 2 , 

F. Gasthuys 1 , P. Verleyen 2 , A. Martens 1 

1 Faculty of Veterinary Medicine - University of Ghent, Department of 

Surgery and Anaesthesiology of Domestic Animals, Merelbeke, 

Belgium 


Medical Imaging of Domestic Animals, Merelbeke, Belgium 

Introduction: Fractures represent one of the most common 

orthopaedic disorders in camelids. Although they are considered 

excellent patients for treatment of orthopaedic injury, fracture 

management can be an interesting challenge for veterinary surgeons. 

Case history, clinical findings and treatment: A 2-month old female 

alpaca, weighing 17 kg, was referred with an acute, non-weight 

bearing, right forelimb lameness. A closed, comminuted, non-articular, 

displaced, oblique fracture of the proximal radius and ulna with a 

dorsolateral butterfly fragment of the radius was diagnosed on 

radiographs. Ceftiofur sodium and carprofen were administered 

preoperatively and continued for 12 days. After sedation and induction, 

anaesthesia was maintained with isoflurane in oxygen. Methadone was 

administered during surgery and continued for 5 days (q 6h). During 

open reduction of the fracture, the butterfly fragment was fixed using 

2.7mm cortical screws with lag screw technique. Application of a 

Camelids 145

circular external skeletal fixator (CESF), “Ilizarov”, was chosen: 3 

complete (1 proximal and 2 distal to the fracture) and 1 half (olecranon) 

aluminium ring(s) were connected by use of 3 threaded connecting 

rods. Under fluoroscopic guidance 2.0mm Kirschner wires were 

placed, secured to the rings and tensioned. Recovery from anaesthesia 

was uneventful. The first day, the alpaca was very painful, but 

radiographic control showed a good fracture repair. After handling the 

alpaca, an acute, non-weight bearing, left forelimb lameness was 

detected and a displaced Salter-Harris type 1 fracture of the apophysis 

of the ulna was diagnosed on radiographs. During a second surgery, the 

fracture was reduced and fixed using a ”tension-band“, wiring 

technique. Despite two fractured and surgically repaired forelimbs, the 

alpaca was able to stand-up alone and walk around within 24 hours 

after the second surgery. Twelve days after the 2 nd surgery, the alpaca 

was discharged from the hospital. Follow-up radiographs showed 

good, normal healing in both forelimbs and intact fixation material. 

After 9 weeks, the CESF was removed under general anaesthesia, but 

the cortical screws and “tension band” were left in place. 

Outcome: The fractures healed without complications and the alpaca 

became completely sound. 

Conclusion: This report shows that bilateral surgical fracture repair 

and the use of “Ilizarov” in alpacas are not a “mission impossible”. 

Key words: alpaca, fracture, circular external skeletal fixator 

143 The Survey of Cystic Lithioasis in Camelids 

M. Zaeemi, Z. Khaki 



Camel is one of the most resistant animals in dehydration and arid 

climate. So it still used in desert area like central and Southern regions 

of Iran. In spite this fact there isn't enough information about camel and 

its diseases. Because of high ability in urine concentration, increase the 

probability urolithiasis in comparison with other large animal, that 

could induce obstruction and rupture in urinary bladder etc. 140 urinary 

bladders of camel were studied (male: 130, female: 10). Urolithiasis 

was reported only in adult, male camel, including two stones about 0/6 

- 0/8 mm diameter and 0/4 gr weight, with rough and hard surfaces. 

Like in other animals they are composed of Ca. Appearance urine 

analysis was similar to normal samples in clearance, pH, color, specific 

gravity. 1. Microscopic analysis: Presence of cells in urinary sediment 

(RBC, WBC, Epithelialcell) wasn't observed significant differences 

with nomal samples. 2. Microbial culture: Candida albicans from 2 

cultures medium and E. coli from 4 of them were separated, but any 

microorganisem was separated from urolithiasis sample. 3. 

Biochemistry tests: Enzymatic activity of ALP, AST and GGT in 22 

samples was measured. Only E. coli infected samples have increased 

activity of AST, ALP without any changes in the activity of GGT.There 

is a significant relation between E. coli infection and increasing of 

AST, ALP activities. Therefore it can be index of E. coli infection. But 

urolithiasis has significant relation with neither presence E. coli nor 

increased activity of these enzymes. Of course because of insufficient 

urolithiasis cases haven't high statistical value. 

Key words: camel, urolithiasis, cystic lithiasis, urinary bladder 

144 Normal Arthoscopic View of the Fetlock Joint in the 

Dromedary Camel 

M. Ali 

Faculty of Veterinary, Department of Surgery, Dokki, Giza, Egypt 

The present study was carried out on the fetlock joint and pastern of the 

dromedary camel for description of its normal arthroscopic view. 

Arthroscopy was performed on four joint specimens and three 

anaesthetized animals to evaluate its efficacy and to study the 

arthroscopic portal of the joint and the problems that could be occurred 

during and after arthroscopy. The joint capsule of the fetlock joint in 

the dromedary camel posses a separate synovial sac for each digit, and 

no communication between the articular cavities of the fetlock joint of 

the same limb was found on the dorsal aspect. Therefore two 

arthroscopical portals (lateral and medial) were required. The most 

suitable site for distention of the joint capsule before athroscopy was 

through the proximal- palmar pouch as the distal palmar pouch was 

small and difficult to be palpated and injected. The site of the puncture 

was at the center of the groove bounded by the collateral branch of the 

M. interosseous medius and large metatarsus at a point of 1 cm 


proximal to the joint. The current study revealed that arthroscopy of the 

fetlock joint through the lateral portal allowed examination of the distal 

end of the metacarpal bone and the proximal end of the first phalanx of 

the digit IV. The medial portal allowed examination of the peripheral 

part of the distal end of the first phalanx of the digit III. For complete 

examination of the joint cavity, two portals (lateral and medial) were 

also required. The palmar portals allow examination of the proximal 

sesamoid bones, the synovial membrane, and the synovial villi. 

Key words: Dromedary Camel, arthroscopy, fetlock joint 

184 Investigations of Endo- and Ectoparasites in Camels 

(Camelus Bactrianus) in the Great Lake Depression of 

Western Mongolia 

D. Zaspel 1 , A. Koehler 1 , R. Sodnomdarjaa 2 , M. Baumann 3 , 

P. Clausen 1 

1 

Faculty of Veterinary Medicine, Freie Universitaet Berlin, Institute 

for Parasitology and Tropical Veterinary Medicine, Berlin, 

Germany 

2 

State Central Veterinary Diagnostic Laboratory, Ulaanbaatar, 

Mongolia 

3 

Faculty of Veterinary Medicine, Freie Universitaet Berlin, 

International Animal Health, Berlin, Germany 

In summer 2004 a study was conducted with the objectives (i) to estimate 

the parasitological prevalence of Trypanosoma evansi in serologically 

positive camels or in camels with clinical signs suspected for surra, and 

(ii) to estimate the level of infection with ecto- and endoparasites other 

than T. evansi in camels with poor body condition or signs of myiasis in 

Chandmani, Dorgon and Myangad, north-eastern Khovd, Mongolia. A 

questionnaire survey was conducted to assess the importance of Bactrian 

camel husbandry in the study region. The average herd size was 19, with 

a minimum of 2 and a maximum of 136 heads. Camels are mainly used for 

production of meat (88%), milk (77%) and manure (7%) and for paying of 

dowry (15%). All camels are used as pack animals and for wool 

production. Trypanosoma evansi could not be detected in any of the 154 

examined camels. This finding corresponds well with the low 

trypanosome antibody prevalences for camels in Chandmani, Dorgon and 

Myangad, as reported earlier. An important number of camels was found 

to be infected with Wohlfahrtiosis (27.9% of all examined animals); most 

of them were females (83.7%) with genital affections (97.4%). 

Wohlfahrtiosis was also detected in somae calves. In adult males (16.3% 

infected), affections were mostly observed at the nasal region (71.4%). 

Affected camels received local treatment with a pyrethroid solution called 

Krilin and systemic treatment with a pour-on formulation of moxidectin 

(Cydectin ® , Fort Dodge, 10 ml/100 kg). 

Key words: camel husbandry, Mongolia, Trypanosoma evansi, genital 

myiasis, Wohlfahrtia magnifica 

185 Seasonal Effects on Morphometric Parameters of Uterine 

Cornua in One-humped Female Camel (Camelus 

Dromedarius) in Pakistan 

Mr. Ali 1 , Dr. Sarwar 2 , Dr. Rehman 2 , Mr. Shahid 1 , Mr. Rehan 1 

1 Agriculture University, Anatomy, Faisalabad, Pakistan 

2 Agriculture University, Animal Reproduction, Faisalabad, Pakistan 

Macro and microscopic characteristics and morphometry of different 

parts of uterine horns were studied in 25 clinically healthy adult female 

one-humped camels (Camelus dromedarius) during four seasons i.e. 

winter (n = 7), spring (n = 6), summer (n = 6) and autumn (n = 6) of 

Pakistan. The tissues were processed by paraffin sectioning technique and 

stained by Hematoxylin and Eosin (H & E). Morphometric analysis was 

done with the help of ocular and stage micrometers. The mean - SE values 

of external length of both left and right horns were 12.73 ± 0.44 cm (8.90 

- 17.20 cm) and 10.31 ± 0.41 cm (7.20 - 16.50 cm), internal lengths were 

15.38 ± 0.56 cm (10.50 - 22.50 cm) and 12.01 ± 0.55 cm (8.40 - 19.3 cm), 

thickness were 0.92 ± 0.02 cm (0.72 - 1.10 cm) and 0.88 ± 0.01 cm (0.72 

- 1.03 cm), circumferences of narrow part were 9.36 ± 0.23 cm (7.50 - 

12.20 cm) and 8.36 ± 0.26 cm (5.20 - 10.30 cm), circumferences of 

middle were 11.13 ± 0.31 cm (8.80 - 14.30 cm) and 9.96 ± 0.29 cm (6.50 

- 12.60 cm), circumferences of uterine end were 13.34 ± 0.41 cm (9.90 - 

17.60 cm) and 11.73 ± 0.33 cm (8.10 - 14.40 cm). The mean - SE values 

of thickness of surface epithelium of left and right horns were 19.3 ± 0.78 

µm (14 - 30 µm) and 18.6 ± 0.85 µm (10 - 27 µm), thickness of glandular 

epithelium were 21.2 ± 0.77 µm (14 - 30) and 20.6 ± 0.65 µm (14 - 26 µm), 

number of uterine glands (per unit area = 1mm) were 10.52 ± 0.52 (6.00 -

1600) and 10.44 ± 0.62 (6.00 - 16.00), diameter of uterine glands were 

55.18 ± 2.31 µm (35.00 - 85.00 µm) and 54.08 ± 1.79 µm (38.00 - 70.00 

µm), thickness of endometrium were 3951 ± 193.26 µm (2250 - 5875 µm) 

and 3864 ± 185.7 µm (2425 - 5800), thickness of myometrium were 

4391.9 ± 166.4 µm (2650 - 5790 µm) and 3981.9 ± 164.8 µm (2650 - 5790 

µm). The gross and microscopic parameters of uterine cornua were 

influence by season and had significant difference in the values during the 

winter and summer seasons. The values of all the parameters studied; of 

both left and right horns; were significantly higher in winter as compared 

to summer season. 

186 Morphometric Studies of Uterine Body and Cervix in Female 

One-Humped Camel (Camelus Dromedarius) during Four 

Different Seasons in Pakistan 

Mr. Ali 1 , Dr. Sarwar 1 , Dr. Rehman 2 , Mr. Shahid 1 , Mr. Rehan 1 

1 Agriculture University, Anatomy, Faisalabad, Pakistan 

2 Agriculture University, Animal Reproduction, Faisalabad, Pakistan 

Macro and microscopic characteristics and morphometry of different 

components of uterine body and cervix of adult clinically healthy female 

one-humped camels (Camelus dromedarius) were done under natural 

ecology of Pakistan. 25 samples of reproductive tract were collected 

during four seasons i.e. winter (n = 7), spring (n = 6), summer (n = 6) and 

autumn (n = 6) of the year. Tissues preparation was done by paraffin 

technique and staining by Hematoxylin and Eosin (H & E). Ocular and 

stage micrometers were used for morphometric analysis. The overall 

mean - SE values of length of body of uterus were 4.87 ± 0.17 cm (3.70 - 

7.70 cm) and of circumference of body of uterus was 15.75 ± 0.43 cm 

(12.50 - 20.20 cm). The mean - SE value of thickness of surface 

epithelium of body of uterus was 18.8 ± 0.71 µm (14.00 - 30.00 µm), 

thickness of glandular epithelium uterine body was 19.6 ± 0.72 µm (14.00 

- 25.00 µm), number (per unit area = 1mm) and diameter of endometrial 

glands of body of uterus was 10.00 ± 0.52 (6.00 - 14.00) and 54.40 ± 2.47 

µm (36.00 - 80.00 µm), thickness of endometrium and myometrium of 

body of uterus was 3810 ± 171.19 µm (2425 - 5800 µm) and 4940.3 ± 

215.1 µm (3348 - 8000 µm). The overall length of cervix was 6.09 ± 0.24 

cm (4.50 to 9.70 cm), circumference was 16.26 ± 0.43 cm (13.40 to 20.70 

cm) and number of cervical rings was 5.20 ± 0.20 (4.00 to 7.00). The 

overall value of thickness of surface epithelium of cervix was 21.42 ± 0.96 

µm (14.00 to 34.00 µm), thickness of tunica muscularis of cervix was 

5368.72 ± 279.12 µm (3348.0 to 8750.0 µm). There was no significant 

difference in the length of body of uterus and of cervix and number of 

cervical rings among the four different seasons of the year but 

circumference of both parts and all microscopic parameters were 

significantly higher in winter as compared to summer season. Gross and 

microscopic structures of the reproductive organs of female camel under 

study had much effect of season. These parameters showed variation in 

the physiological sexual behavior during the breeding and non-breeding 

seasons of female one-humped camel in Pakistan. 


844 Anatomical Study of Stifle Joint in Iranian Camel (Camelus 

Dromedarius) 

Mh. Yousefi 

School of Veterinary Medicine, Semnan University, Basic Sciences, 

Semnan, Iran 

The stifle joint is the most elaborate synovial (diarthrosis) joint and 

important in animals locomotion, because the pelvic limbs are acting 

propel the body. To study the anatomical structure of camel stifle joint, 

six adult camels stifle joints were collected from slaughterhouse. 

Radiograghs were taken from specimens before and after injection of 

sulfate barium. Then specimens were fixed in 10% formalin solution 

and were dissected. This study showed that the camel’s stifle joint 

consist of three articular sacs; femoropatellar, lateral femorotibial and 

medial femorotibial sac. These articular sacs in camel stifle joint were 

communicated with each other. Lateral and medial collateral ligaments 

and intracapsullar (meniscal and cruciate) ligaments were found in this 

joint. The loop or sheath in the medial part of patella, made by 

parapatellar cartilage and medial patellar ligament, seemed to be weak 

compared to the equine. 

Key words: camel „anatomy”, stifle joint 

845 The Influence of Age, Sex and Altitude on the Morphometry 

of Red Blood Cells of the Dromadary Camel 

M. Melizi, N. Adili 

Batna University, Department for Veterinary Medicine, Batna, 

Algeria, Algeria 

Aim: This study has been conducted on the dromadary camel of the 

chaambi breed in order to reveal the influence of age, sex and altitude 

on the morphometry of red blood cells. 

Materials and methods: The study has been performed on a dromadary 

camel population of the chaambi breed. 

The population has been divided as follows: – G1 15 adult males; 

– G2 07 young males; – 07 young females; – G3 15 adult non pregnant 

females. 

Each two groups live at a different altitude region: 

– Region A: Wilaya of Biskra at 110 metres above the sea-level; 

– Region B: Wilaya of El-Oued at or under the sea level (zero metre); 

Blood samples were obtained by venopuncture of the jugular vein. 

Blood smears have been executed on glass slides immediately after 

sampling.The smears were colored according to the May-Grunwald- 

Giemsa method.The morphometric study was achieved using with a 

graduated ocular and a micrometric slide with an immersion optic 

microscope at a grossissement of 100. The statistical analysis was 

undertaken using the T student test 

Results: The results did not show any significant influence of age on 

the morphometry of red blood cells between the young male and female 

dromadary camels of the two regions. Indeed, sex did not have any 

influence on the morphometry of red blood cells between young and 

adult dromadary camels. However, concerning the altitude, the study 

showed a significant difference between the wilaya of El-Oued group 

and the wilaya of Biskra.The RBC of the adult dromadary camels of the 

wilaya of El-Oued are significantly larger. 

Conclusion: The morphometric study of the RBC of the dromadary 

camels that had been achieved on 88 blood smears in order to study 

the influence of age, sex and altitude showed: 1) The age and the sex 

did not have any influence on the morphometry of the red blood 

cells; 2) The altitude had a significant influence expressed by an 

enlargement of the red blood cells of dromadary camels living at low 

altitude. 

846 A Survey on Frequency of Nephrolithiasis in Camelus 

Dromedarius and Determination of Calculi Composition in 

Najafabad Abattoir 

M. Pourjafar 1 , A. Ansarian 2 

1 School of Veterinary Medicine, Shahrekord University, Clinical 

Sciences, Internal Medicine, Shahrekord, Iran 

2 Faculty of Veterinary Medicine, Islamic Azad University, Shabestar 

Branch, Vetrinary Faculty, Shahrekord, Iran 

Urolithiasis is a metabolic disease of cows, sheep and goats and only a 

few cases are reported in camel. Camel is an animal which 

anatomically and physiologically adapted itself to weather condition of 

desert and also it can produce urine which its viscosity is twice more 

than viscosity of the sea water. Some of the factors for producing 

urinary calculi are high viscosity of urine, water deprivation and urine 

retention. In this survey the rate nephrolithiasis among camels 

slaughtered in Najafabad abattoir was investigated and to do this, the 

kidneys of 389 camels in four different range ages 1-3, 3-5, 5-7 and 

more than 7 years old were examined. Among these groups 315 camels 

were male and 71 ones were female. Survey showed a urinary calculus 

in the right kidney of a 5-7 years old male camel .It was a cream stone 

with 2 by 2 by 2 mm dimensions, 0.6gr weight and smooth surface. So 

the rate of outbreak of disease is 0.26 percent.Chemical ingredients of 

this urinary calculus were as follows: Calcium carbonates 43%, 

Calcium hydrogen phosphate 35%, ammonium magnesium phosphate 

20%, and other ingredients 2%, so this urinary calculus might be 

known as calcite (calcium carbonate. (It must be emphasized that 

despite the high viscosity of camel's urine, the rate urinary calculi are 

very rare.This finding may be explained and related to the following 

factor: 1. Ad libitum access to water in modern raising methods in the 

area of this study; 2. Castration of camels is not performed in this area; 

3. High diameter of urinary tracts (pelvis and ureter) of camel; 4. High 

concentration of salts in the rations of camel; 5. High ratio of roughages 

to concentrates in the rations of camel. 

Key words: camel, calcium carbonate calculi, Najafabad, urolithiasis 

Camelids 147

847 Llama Anesthesia for Orthopedic Surgery 

R. Godoy, R. Almeida, C. Leite, C. Pereira, L. Gouvea, FH. Ximenes, 

L. Gontijo, JR. Borges, R. Ferreira II 


Objectives: Describe the anesthesiologic protocol used for an 

orthopedic procedure on a llama. 

Materials and methods: One male llama, one year and a half old was 

admitted to the University of Brasilia’s veterinary hospital to be subjected 

to a surgical procedure of fracture reduction. The drugs used for preanesthetic 

medication were the butorphanol (0.05mg/kg) and romifidine 

(0.04mg/kg), administered by intramuscular route. After trichotomy and 

anti-sepsis of the operative field and the lumbosacral region, an anesthetic 

button was made with lidocaine 2% without vasoconstrictor at the point 

where the puncture would be performed. The animal was placed in the 

right lateral recumbency, with its head high in relation to the body and 

subject to puncture of the spinal subarachnoid space with 18 Gauge 

needle, and administration of ropivacaine (0.75mg/kg). About two hours 

after the sedation, there was need of anesthetic rescue with ketamine 

(1.0mg/kg) associated with diazepam (0.1mg/kg). The protocol was 

repeated after 25 minutes. After another 25 minutes, it was necessary to 

administrate a new dose of romifidine (0.04mg/kg). At the end of the 

surgical procedure, phenylbutazone (4.0mg/kg) was administered. Then 

an epidural catheter was placed, through which romifidine (0,05mg/Kg) 

was administered, diluted in saline solution to the volume of 5 ml aiming 

postoperative analgesia. 

Results: The animal showed satisfactory answer to the protocol of 

sedation, being observed only sialorréia as side effects. Soon after 

subarachnoid anesthesia transient dyspnea and tachycardia were 

noticed. The sensory and motor blockade was effective and led to the 

completion of the surgical procedure without the need of using general 

anesthesia techniques. Regarding anesthetic rescue, the only adverse 

effect observed was slight agitation of the animal. Immediately after 

the administration of phenylbutazone, the animal showed tachycardia, 

tongue hypotonia and catalepsia being treated with dexamethasone. 

Conclusion: The anesthesiologic protocol was effective for the 

completion of the surgical procedure, demonstrating hemodynamic 

stability, satisfactory analgesia and few complications. The 

dexamethasone was effective in treating shock, promoting the recovery 

of the consciousness of the animal. Nearly four hours after the end of 

surgery the animal came to death by respiratory complications that can 

be attributed to the episode of anaphylactic reaction. 

Key words: camalid, fracture, anaphylatic reaction 

848 Treatment of Long Bone Fractures in Llamas: Study of Two 

Cases 

R. Godoy, C. Pereira, C. Leite, FH. Ximenes, L. Gouvea, L. Gontijo, 

R. Ferreira II, A. Teixeira Neto, JR. Borges 


Objectives: Report of two cases of long bone fractures in llamas with 

different treatments. 

Materials and methods: In University of Brasilia’s Veterinary 

Hospital were assisted in 2007, two llamas, males, both presenting 

complete fractures. The first animal suffered a metatarsus fracture of 

the right hind limb, while in the other the involved bone was the tibia of 

the left hind limb. The methods used were respectively, conservative 

treatment to reduce the fracture using synthetic cast and polyvinyl 

chloride (pvc) splint and surgical treatment using external hybrid 

fixation. For the surgery, the second animal was submitted to 

tranquilization and subarachnoid anesthesia and the development of 

treatments has been monitored by radiographic examinations. 

Results: Soon after the immobilization the first animal could already step 

with the affected member. Radiography was performed, in which it was 

observed an efficient reduction, juxtaposition of the bone fragments. The 

animal was discharged after thirty days, presenting satisfactory 

consolidation of fracture. In the second case, the placement of splint was 

also tried, however it was unsuccessfull due to the anatomical location 

accompanied by the difficult temperament of the animal, which required 

the surgical procedure. This has led to a stabilization of bone fragments 

and neutralization of active forces in the fracture. The results of such 

reduction on bone consolidation could not be acessed as the animal came 

to death hours later due to post-anesthetic complications. 

Conclusion: Both methods have proved effective in the correction of 

fractures in llamas. The first method may be indicated for reduction of 


fractures of distal long bones, offering low cost and not requiring 

sophisticated equipment for their achievement what makes it possible 

to be used in the field. The second method use is indicated in complex 

fracture cases, located in bones covered by developed musculature 

where the simple imobilization is not enought to neutralizate the 

fracture forces. It’s also important to know that it is a hight cost 

technique that needs trained professional and a equiped surgical center. 

Key words: camalids, external hibrid fixator, cast, long bone 

849 Camel Pox in Saudi Arabia: Retrospective Study 

T. Fouda, A. Bakhsh 

College of Veterinary Medicine and Animal Resources, Kfu, Clinical 

Studies, Houfof, Saudi Arabia 

Pox had been studied by several authors in different animal species in 

different countries. Camel pox was first recorded in young camels (Leese, 

1909). The earliest identification of this disease by veterinarians in Saudi 

Arabia was 1958 (Anon, 1963) and was described as dermatitis. Camel 

pox was identified & isolated and designated officially by Hafez et al. 

(1986). Subsequently, different forms had been described in different 

locations. Consequently this article describes the different clinical aspects 

of camel pox and its identification in Saudi Arabia. 

Key words: camels, pox lesions, Saudi Arabia 

850 Study of the Urethral Glands Presence in Distal Extremity of 

Penis in One Humped Camel (Camelus Dromedarius). 

Mh. Yousefi 1 , H. Gilanpour 2 , P. Tajik 3 

1 School of Veterinary Medicine, Semnan University, Anatomy, 

Histology and Embryology, Semnan, Iran 

2 Faculty of Veterinary Medicine, University of Tehran, Anatomy, 

Histology and Embryology, Tehran, Iran 

3 Faculty of Veterinary Medicine, University of Tehran, 1- 

Department of Clinical Medicine, Tehran, Iran 

The one part of male genital system is genital glands. These glands on the 

basis of exist reports are including, prostate gland, bulbourethral gland 

and urethral glands in pelvic urethra. To accomplish this research, five 

histological specimens from five camels were preparated in 

slaughterhouse. These specimens were fixed in 10% formalin solution. 

Routine histological procedures were made by an autotechnicon paraffin 

blocks of histological speciments from distal extremity of penis and were 

sectioned at 6 microns, Stained with Haematoxylin and Eosin and studied 

under light microscope. In conclusion the studied of histological sections 

were showed that urethral glands in addition to pelvic urethra, existed in 

wall of distal extremity of penile urethra in the neck of camel’s penis. Also 

two mass glands of penile urethra were distinguished. One mass was 

smaller and situated on the lateral aspect of urethra just to the corpus 

cavernosum of glans of penis, this gland was situated on the dorsal surface 

of urethra before inclined urethra to left side. And other one mass was 

greater and attached to the ventral surface of the urethra. This gland was 

situated on the dorsolateral of urethra before inclined urethra to left side. 

The round and elliptical and long sections of this mass glands were 

consist of high cuboidal or columnar cell layer. These cells had a round 

basically nucleus and cytoplasm similar to white color. 

These tubular glands may have ability to secret serous substances. 

Key words: camel, anatomy, penis, urethra 

851 Camel Anesthesia for Orthopedic Procedure 

R. Almeida 1,2 , C. Leite 1 , A. Teixeira Neto 1 , S. Maguilnik 1 , 

A. Farias 2 , R. Godoy 1 

1 Universidade de Brasília, FAV, Brasília-DF Brazil, Brazil 

2 UPIS Faculdades Integradas, Departamento de Medicina 

Veterinaria, Brasília-DF Brazil, Brazil 

Objectives: Describe the anesthesiologic protocol used for an 

orthopedic procedure on a camel. Materials and methods: A female 

camel was anesthetized at the Brasilia Zoo for an washout followed by 

chemical arthrodesis of the tarsometatarsal articulation. The animal 

was submitted to food and water fast respectively 24 and 12 hours. It 

was sedated with xylazine (0.2 mg / kg) by intramuscular route (IM) 

and ten minutes later, induced with cetamine (5.0 mg / kg IM). After 

venopuncture with a catheter, the animal was submitted to total 

intravenous anesthesia (TIVA) using a combination of guaiafenesin in 

the dose of 50 mg / kg / h associated with cetamine (1.0 mg / kg / h) and

Xylazine (0.1 mg / kg / h). 35 minutes after the start of TIVA the animal 

showed signs of being in a superficial stage and the anesthesia was 

complemented with diazepam (0.05 mg / kg) associated with cetamine 

(0.5 mg / kg) intravenously. This association was repeated twice more 

during the TIVA 30 and 60 minutes after the first dose. At the end of the 

procedure, a epidural catheter was placed in the space between the first 

and second coccigean vertebras through which morphine was 

administered at a dose of 1.0 mg / kg once a day for three days. 

Throughout the anesthetic period heart and respiratory rates were 

monitored and also periferic oxygen saturation and protective reflexes. 

The anesthesiologic procedure lasted 2 hours and 45 minutes. 

25 Concept, Implementation and Evolution of Uniform, a 

Veterinary-Based Herd Health and Production Management 

Program for the Dairy Veterinary Practice and Farms 

B. Van Ranst 1 , T. Caluwaerts 2 , HJ. Van der Beek 1 , G. Opsomer 2 

1 

Uniform, Assen, Netherlands 

2 

Ghent University, Dep Reproduction, Obstetrics and Herd Health, 


To fulfil the current (and future) consumers: wish to have high quality 

milk for a low price at their disposal, farmers will need to significantly 

adapt their business. Herds will have to grow continuously and a 

further selection of cows with an ever increasing genetic potential to 

produce milk will be necessary to remain competitive. High yielding 

dairy cows however need significantly more attention in terms of cow 

comfort, animal welfare and disease control. To protect the animals 

from major infectious diseases on the one hand and to safeguard the 

milk from being contaminated with pathogens on the other hand, herds 

will furthermore need to possess strict prevention and control protocols 

based on regularly carried out surveillance and monitoring programs 

including full traceability of animals, feed, the use of drugs etc. The 

veterinarian will play a crucial role herein. While farmers need to be 

more and more qualified to manage their high yielding cows, their time 

management becomes more and more critical in the ever growing 

herds. Therefore, in modern herds a steeply growing automation is 

currently going on. Mobile data collection, increased use of on-farm 

sensors (activity- and milk meters, in-line milk analysis) and a higher 

automation rate in milking parlours start to revolutionize working 

routines and protocols of both vets and farmers. Besides the automated 

collection of data, their centralisation and interpretation is evenly 

important. New internet features like dairy data warehouse are 

therefore becoming more and more popular and maximize the (re-)use 

of all timely gathered data. Overall globalization furthermore 

stimulates benchmarking on a worldwide level and necessitates farmers 

and veterinarians from all places in the world to be able to 

communicate and interact with each other. The Uniform dairy herd 

health and production program will be presented in terms of the above 

mentioned points. Concepts, methodology and results will be 

discussed. The program actually runs on more than 6.750 dairy farms in 

more than 25 countries. 

Key words: dairy, herd health, automation, software program 

26 Lactation Curve Analysis in Dairy Herd Management 

M. Krogh, C. Enevoldsen 

Faculty of Life Sciences, University of Copenhagen, Department of 

Large Animal Sciences, Copenhagen, Denmark 

Milk yield at herd and lactation level have been used to compare 

herds and cows for decades. The total milk production during a 

lactation is financially important. However, milk production in early 

lactation is less efficient than later in lactation and the milk 

production often is affected by disease. Consequently, important 

information about efficiency and health can be derived from the 

profiles of milk production at lactation and herd level (“lactation 

curves”). The objective of this study was to provide a set of reference 

values that allows us to perform “lactation curve benchmarking”. We 

modelled milk production within herds for calvings during the last 2years. 

There were 6-11 milk recordings per year per cow and milk, 

fat and protein were transformed into Energy Corrected Milk 

(ECM). We specified a random coefficient regression model with 

Herd Health Management and Epidemiology 

Results: The animal showed satisfactory answer to the protocol of 

sedation, being observed almost no side effects. Soon after 

beginning the TIVA transient respiratory depression was noticed. 

The respiratory rate decreased form 17 (baseline) to 7. This 

depression lasted 25 minutes and then the parameter returned to 

baseline values. There were no significant changes at heart rate and 

oxygen saturation. 

Conclusion: The anesthesiologic protocol was effective for the 

completion of the procedure, demonstrating hemodynamic stability, 

satisfactory analgesia and no complications. 

Key words: camalid, total intravenous, TIVA 

piecewise linear effects of stage of lactation (DIM). Both fixed and 

random effects of “peak” at 60 DIM, slope before 60 DIM 

(acceleration) and slope after 60 DIM (persistency) were estimated 

separately for first, second and later lactations (3 fixed and 3 random 

effects per parity group). From the covariances we estimated 

correlations between peak and slopes. This baseline model 

consequently allows us to monitor the major characteristics of the 

lactation curves within herd and within parity group. Estimates were 

derived from 542 similar herds (Danish Holstein cows). The table 

below shows the distribution of estimates from third lactation and 

older. The data shows that there is a large proportion of herds where 

the slope before 60 DIM is steeper than after 60 DIM. Such 

“atypical” lactation curves may be indicators of some major 

management or health problem. The variances of the slope estimates 

represent the uniformity of the cows within herd. This may also be 

an important and objective health indicator as poor management and 

health probably will increase variance. Correlations between peak 

and persistency may indicate efficiency. In conclusion the estimates 

provide valuable information about achievable goals for lactation 

curve characteristics. 

Lactation curve 25%-percentile Mean 75%-percentile 

characteristics 

(ECM) 

Peak at 60 DIM 35.4 37.8 40.2 

Change from -1.7 -0.1 1.2 

10 to 60 DIM 

(„acceleration”) 

Change from -16.5 -14.7 -12.9 

60 to 305 DIM 

(persistency) 

Variance of peak 29.8 42,2 50.4 

Variance of 30.2 47.3 61.3 

acceleration 

Variance of 33.2 49.9 61.8 

persistency 

Correlation -0.65 -0.53 -0.43 

between peak 

and persistency 

27 Developing and Marketing a Beef Production Medicine 

Program in Private Practice 

W. Hilton 

Purdue University, School of Veterinary Medicine, Veterinary 

Clinical Sciences, West Lafayette, United States 

Beef cattle veterinarians are being asked more frequently to not only 

aid in the prevention of disease in the herds they service, but are also 

being asked to assist in improvement of herd production and ultimately, 

herd profitability. As our roles evolve, we must add these production 

medicine and herd health services to our job description if we truly 

want to assist the producer in growing a successful business. While 

many veterinarians provide herd health services to their dairy, beef 

feedlot and swine herds, doing so in a beef cow-calf herd is a relatively 

new idea. We will explore not only how to develop this program in a 

private practice, but also how to market these services to the benefit of 

your clients. 

Key words: beef production medicine, herd health, consultation 

Herd Health Management and Epidemiology 149

28 Preliminary Results of a General Health Monitoring System 

for Cattle in Austria 

W. Obritzhauser 1 , C. Egger-Danner 2 , B. Grassauer 1 , W. Holzhacker 1 , 

P. Winter 3 

1 

Chamber of Veterinaries, Vienna, Austria 

2 

ZuchtData EDV-Dienstleistungen GMBH, Vienna, Austria 

3 

University of Veterinary Medicine, Clinic for ruminants, Vienna, 

Austria 

A project to establish an Austrian wide health monitoring system for 

cattle is currently being implemented. An overall aim of the project is 

the development of a genetic evaluation for health traits for the main 

Austrian cattle breeds. Within the project diagnostic data, which have 

to be documented by law by the veterinarian (law of drug control) are 

standardised and recorded into a central database. All farms under 

performance recording are free to join the project. Responding to an 

increasing interest in health issues, already 10,500 farms have been 

participating by the beginning of the year 2008 with more farms 

continuously joining. To increase the health status of the animals by 

management measures health reports are elaborated and provided to 

farmers and veterinarians. Validated diagnostic data provided by 

veterinarians who are joining the project from the beginning and 

delivering complete data sets are included in the study. In total, 

recorded diagnostic data from 556 dairy farms (average farm size 20.9 

cows) can be evaluated. The main problems in dairy farms are caused 

by reproduction diseases and mastitis, which require intervention by 

the veterinarian. The average calving interval of the farms included is 

398 days. In case of mastitis the average somatic cell counts in 

primiparous cows are 117.000 cells per ml and in multiparous cows (2 

lactations) 216.000 cells per ml, respectively. First experiences show 

that many diagnostic data are recorded in farms at a very early stage of 

disease. The monthly provided health reports support successful and 

economical farm management and are important tools for a good 

cooperation between farmers and veterinarians. 

Key words: health monitoring, health reports, farm management 

58 A Contrast of Management Practices in Beef Herds with High 

versus Low Conception Rates 

C. Heuer 1 , D. West 2 , G. Tattersfield 2 , R. Jackson 1 

1 Massey University, EpiCentre, Palmerston North, New Zealand 

2 Massey University, Institute of Veterinary, Animal and Biomedical 

Sciences, Palmerston North, New Zealand 

Objectives of study: The study aimed to identify associations between 

management practices and pregnancy rates of beef cattle herds in New 

Zealand. 

Materials and Methods: Veterinary practices with beef cattle clients 

were contacted to provide records of pregnancy testing in two 

subsequent years (2005-06). A questionnaire was subsequently mailed 

to all participating farmers inquiring information about the 

management of beef cow fertility. This information was regressed on 

the proportion of cows pregnant 1-2 months after the end of mating 

using multi-factorial logistic regression adjusting for the possible 

confounding effects of herd size, farm surface area, stocking rate, and 

region. 

Results: Pregnancy data were available from 1217 farms from across all 

regions and agro-ecological zones of New Zealand. From 718 (59%) 

questionnaires that were returned, 150 were excluded because they had 

changed locations or given-up-farming, but also for insufficient data 

entry. The remaining 568 (47%) farms were included in the final analyses. 

Safety and topography of calving paddocks was identified as an influential 

risk factor for conception rates. Too few labour units had negative effects 

on fertility but while this outcome is highly plausible, it does not easily 

translate into a realistic solution. Farmers not using a mating ability test of 

service bulls (70%) achieved similar pregnancy rates but purchased more 

breeding bulls relative to herd size. A ratio of more than 50 cows per 

breeding bull was associated with declining pregnancy rates. Pregnancy 

rates in herds with pure Hereford, Angus and their crossbreeds were 3-6% 

lower than herds with other or mixed breeds. Feed restriction in late 

pregnancy in all ages but especially in heifers was associated with lower 

pregnancy. Temperament of breeding bulls was an indicator of high 

fertility and that The data suggested that culling bulls with -bad 

temperament- would reduce beef cow fertility. 

Conclusions: The study provided insight into management practices 

associated with higher pregnancy rates. These were related to safety of 


pastures, breed, nutrition and the use of a mating ability test for 

breeding bulls. 

Key words: beef cattle fertility, management, cross-sectional study 

59 Comparison in Swiss Dairy Farms with and without a Herd 

Health Program 

M. Hassig 1 , D. Kemper-Gisler 1 , A. Liesegang 2 , U. Braun 1 

1 

Vetsuisse-Faculty Zurich, Department of Large animal clinical 

Sciences, Zurich, Switzerland 

2 

Vetsuisse-Faculty Zurich, Department of Physiology, Zurich, 


The goal of this retrospective study was to compare Swiss dairy farms 

with and without a herd health program by means of really invested 

money. The major goal was to improve fertility. Over seven years from 

1999 to 2005, matching 11 farms with and 11 farms without a herd 

health program were evaluated. All farms were visited on a regular 

basis by the ambulatory field clinic of the Vetsuisse-faculty of the 

University of Zurich. All actions were registered by Oblon Data and 

analyzed by StatView 5.1 by means of analysis of variance with 

repeated measures. About 130000 records were analyzed. This study 

showed that dairy farms with a herd health program spent less money 

per animal per year than farms without herd health program. The 

money spent for routine visits as well as emergency visits was not 

different. Also the money spent for medication did not differ. The costs 

for veterinary work alone did not alter between dairy farms with and 

without a herd health program. It was not possible to prove a financial 

advantage for farms with a herd health program compared to dairy 

farms without a herd health program based on the smaller financial 

investment due to lesser veterinary costs. 

Key word: herd health program, financial cost, fertility 

60 Rearing Conditions Affect Lifetime Performance and Health of 

Dairy Cows: an Observational Study in Swedish Herds 

J. Hultgren, C. Svensson 

Swedish University of Agricultural Sciences, Department of Animal 

Environment and Health, Skara, Sweden 

To study associations of rearing conditions with culling, calving interval 

(CI), milk production, and veterinary-reported clinical mastitis (CM) 

throughout productive life, data were collected from 2127 cows (49% 

Swedish Reds, 49% Swedish Holsteins), representing all female 

animals born 1998 in 110 herds in southwest Sweden. Median length of 

productive life (LPL) in culled cows was 780 d (14% of records 

censored). The Weibull proportional hazards model showed that a 

calving age of 28 to 31 mo was associated with the highest culling risk, 

1.2-fold higher than calving at 7 calves from 3 to 7 mo was associated with a 1.7-fold 

increase in risk, relative to litter pens. If a cow had changed housing 

system 4 times before 1st calving it increased the risk of culling 1.4 

times, relative to 2 housing changes. Pregnancy status, CM, and milk 

yield all affected culling dramatically. The risk of CM was studied by a 

generalised linear mixed model. In a given lactation, a cow was defined 

as affected by CM if one or several cases were reported by a veterinarian 

starting from 7 d precalving. The mean CM incidence was 14% per 

lactation; 31% of the cows had CM at least once during their productive 

life. Of the variation in CM risk, 90% was due to factors at the lactation 

level. Severe diarrhoea between 3 and 7 mo of age was associated with 

2.8-fold higher odds than moderate diarrhoea during the same period. 

The odds changed curvilinearly with prepubertal growth rate, with a 

maximum at 859 g/d, and increased linearly with concentrate ration the 

last 2 mo before 1st calving. Median CI was 381, 380, and 377 d in 

parities 1, 2 and >3, respectively. The applied mixed model (3542 

records in 1550 cows) showed that severe respiratory disease between 3 

and 7 mo of age in Swedish Reds increased CI by 12%. Of the total 

variation in CI, 9.5% was due to herd factors and 12% to cow factors, as 

opposed to variation between lactations. Milk production was studied 

by a mixed model. Estimated mean (SD) production was 9209 (±1428) 

kg ECM/cow-yr. A high prepubertal growth rate, calving for the first 

time in late autumn or winter, a high calving age, and a large increase in 

concentrates around calving increased production. The results are 

indicative of effects of rearing conditions on the lifetime performance of 

dairy cows, likely to affect profitability.

61 Intensive Care - a Prerequisite to Controlling the Transition 

Period in Dairy Cows 

M. Skrivánek 1 , S. Slosárková 2 , P. Fleischer 2 , J. Doubek 1 

1 University of Veterinary and Pharmaceutical Sciences Brno, 

Department of Physiology, Brno, Czech Republic 

2 University of Veterinary and Pharmaceutical Sciences Brno, Clinic 

of Ruminant Diseases, Brno, Czech Republic 

The transition period is the most important phase of the re/production 

cycle. There occur a number of interrelated diseases, participating in 

the so-called syndrome of periparturient crisis of dairy cows. The aim 

was to evaluate the influence of intensive care of the animals during 

this period on their health and re/production parameters. Two groups of 

Holstein cows (n=2x15) within a 400 cows were evaluated. Following 

parameters were used to check the state of health during the transition 

period: 1) clinical score, 2) biochemical score, 3) milk score (always 1- 

5 scores, 1 = physiological state, 5 = severe alteration), and 4) total 

score (3-15 scores). Group I was control, group II was supplied with 

Fortis additive products 21 days a. p. and 7 days p. p. (containing crude 

fibre, energy, vitamins, macro- and micro-elements). The clinical score 

was determined on the basis of BCS, temperature, urine examination, 

digestive, reproductive and locomotion apparatus and mammary gland 

evaluation. Biochemical score was determined based on blood 

examination on days 1, 10 and 25 p. p. Serum samples were examined 

for total proteins, Ca, P, Mg, AST, NEFA and BHB. Milk score was 

determined using milk yield resp. fat and protein contents until resp. on 

the 1 st yield test. Comparing the group I with the group II following 

significant differences (p

Israeli commercial dairy herds consisting of 251-824 high producing 

Israeli Holsteins. Data were collected between March 2006 and August 

2007 and included health, production, reproduction and management data 

and over 250,000 daily BW measurements from 2,167 cows. Time series 

analysis was performed whereby each series of BW measurements was 

represented as the sum of a trend component, periodic component and 

"white noise". In order to construct "standard" relative BW curves, 

estimates from mixed models fit to the data were obtained. Seven day and 

21 day cycles in BW were present in 247 (11.4%) and 715 (33.0%) of the 

cows, respectively. Presence of 21 day cycles was associated with a 

reduction of 33% in the risk of being diagnosed with inactive ovaries. 

Mean days from calving to nadir BW were 37, 54 and 61 for first, second 

and older parity cows, respectively. Mean loss in BW (kg) from calving to 

nadir was 46.5, 68.2 and 78.3 for first, second and older parity cows, 

respectively. The "average" first parity cow lost 6.5% of its calving BW as 

opposed to 8.5% in the older cows. Gain in BW following the nadir value 

was markedly more rapid in first parity cows. The presence of 21 day 

cycles in BW is most probably related to the sexual cycle, as indicated by 

the relationship with inactive ovaries. Monitoring the presence of these 

cycles could be useful for indirectly assessing ovarian activity in dairy 

herds. Compared to older cows, fewer days from calving to nadir BW and 

smaller BW loss from calving to nadir, coupled with a faster post-nadir 

increase in relative BW in first parity cows indicate a smaller energy 

deficit in early lactation in this group. "Standard" relative BW curves for 

the first 120 days in lactation are presented. 

102 Use and Abuse of a Uterine Scoring System in Clinical 

Practice 

D. Nielsen, L. Jensen, R. Jespersen, C. Enevoldsen 


Large Animal Clinical Sciences, Copenhagen, Denmark 

Valid and precise clinical metritis diagnoses are crucial for rational 

decisions on treatments of individual cows, effective monitoring of the 

herd and advanced multifactorial epidemiological analyses within and 

between herds. A steadily growing proportion of Danish cattle 

veterinarians examine and score the uterine status of cows 5-12 days post 

partum on an ordinal 0-9 scale. In addition, the veterinarians record 

whether they treat the cow for metritis in connection with the scoring. 

The objectives of this study were 1) to evaluate the current usage of the 

uterine score as a treatment guideline and 2) to examine the agreement 

between veterinarians on the definition and use of the score values. By 

means of semi-structured telephone interviews we investigated how 20 

cattle veterinarians decided which cow should be treated for metritis. The 

telephone interview showed that the veterinarians used the scoring scale 

as an integrated part of a wide range of other diagnostic criteria and 

anamnestic information. It was an important finding that the 

veterinarians initially gave a concise description of how they used a scale 

value as a threshold for treatment. But after more in-depth questioning 

80% reconsidered and described more complex applications of the scale. 

Rectal temperature was seldom used as a treatment criterion. Such 

implicit treatment strategies may explain some of the interactions and 

apparent inconsistencies revealed by purely quantitative studies of large 

data files with routinely collected data (e.g. the Danish National Cattle 

Data Base). Examples will be shown. To elaborate further on the use of 

the scoring scale independent observations were made by a trained 

practicing veterinarian (>20 years in practice) and an untrained (student) 

on 45 cows 5-12 days in milk (DIM). The results showed good 

agreement (weighted kappa 0.8 [0.72; 0.89]). However, when agreement 

between 2 different observers (student and first author) on the score 

components (odour and quantity) were evaluated in 25 cows 5-20 DIM, 

poor results were obtained (weighted kappa 0.37 [0.01; 0.74] and 0.37 

[0.15; 0.59], respectively. Both studies indicate that further 

communication about treatment strategies based on uterine scores and 

calibration of the scale are needed to obtain a data quality that allows 

valid and precise conclusions about the historic development of uterine 

status and permits multifactorial analysis of the effects of uterine status 

and metritis treatment. 

103 A Government Subsidised Herd Health Pilot Programme: 

Four Years Progress (2004 -2008) 

P. Mullowney 1 , D. Barrett 1 , R. Fallon 2 , J. Egan 1 , M. Good 1 

1 

Department of Agriculture, Fisheries and Food, Training Section, 

6E, Dublin, Ireland 

2 

Teagasc, Grange, Co. Meath, Ireland 


To evaluate the success of a Government Subsidised Herd Health Pilot 

Programme established in 2004. 

Methods: The Programme commenced in January 2004 with a series 

of seminars for participating veterinarians on IBR, BVD, Johne’s 

Disease, Epidemiology and Risk Analysis. These veterinarians 

enrolled 78 of their clients in the scheme. Forty six of these were dairy 

herds, 21 suckler and 11 mixed enterprise. Sixty one herds took part in 

Johne’s control, 18 in BVD control and 8 in IBR. Of the 61 Johne’s 

herds 39 thought that they were free of the infection and wanted to 

prevent its entry whilst the remaining 19 knew they had the disease and 

wanted to eliminate it. A fee was paid to participating veterinarians for 

a risk analysis in each of the herds and testing costs were subsidised in 

the laboratory. Follow up seminars were given to the enrolled 

herdowners and their practitioners in August 2005 and December 2007. 

The scheme was also subsidised by contributions from farm and 

veterinary organisations including Veterinary Ireland, University 

College Dublin, Irish Cattle Breeders Federation, Teagasc and Irish 

Farmers Association. The enrolled herds have been testing for various 

lengths of time. Fifty seven herds have taken blood samples, 14 of 

which have been negative on all tests and 43 having positive samples 

on one or more tests. Twenty herds carried out faecal sampling. 

Fourteen of these had all negative results, while positive samples were 

found in six herds. An annual risk evaluation was carried out in all 

herds to ensure that the client had carried out agreed 

recommendations. Participants have requested a form of certification 

for those herds with all negative results. This would be combined with 

an assessment that the herd is closed. 

Key words: Johnes, IBR, BVD, herd health 

104 A Comparison of Production, Calving and Fertility Related 

Events in 10 Irish Dairy Herds Located in the Vicinity of a 

Chemical Industrial Complex and 10 Dairy Herds Located in 

Non Industrialised Areas 

J. Buckley 1 , P. O'Mahony 1 , J. O'Donovan 2 , D. Berry 3 , K. O'Farrell 4 

1 

Cork County Council, Veterinary Department, Cork, Ireland 

2 

Faculty of Veterinary Medicine, Veterinary Department, Dublin, 

Ireland 

3 

Teagasc, Moorepark, Dairy Production Research Centre, Fermoy, 


4 

Veterinary Consultant, Dairy Production, Cork, Ireland 

Production, calving and fertility data for the years 2001 to 2004 inclusive 

was collated for 10 dairy herds located in the vicinity of a complex of 

chemical industries in the Cork harbour region (target herds) and 10 herds 

located in rural, non-industrialised areas (control Herds). Average milk 

yield per cow, stocking rates and culling rates were similar for the two 

groups of herds. The prime reasons for disposal of animals from both 

groups of herds were infertility, ”old age”, mastitis, lameness and low 

production. The proportion of deaths recorded in both groups was similar 

(1.8% of total calvings) for the four year period. Overall, the proportion of 

male calves born (52%) was significantly greater than females. There was 

no significant difference between groups in the sex ratio, the incidence of 

calving difficulty or the incidence of retained placentae. A higher 

proportion (P

corona virus (BCV) in dairy herds in two neighbouring veterinary 

districts in Sweden, and to explore the association between antibody 

status and some herd characteristics. Forty-one herds were sampled 

from district A and 38 from district B. A pooled milk sample from 

five primiparous cows and bulk tank milk (BTM) was obtained from 

each herd and analysed for IgG antibodies to BRSV and BCV with 

an indirect ELISA. The location of the herds was mapped using a 

geographical information system. Data on herd size, artificial 

insemination (AI) strategy, breed and milk production were accessed 

from the Official Milk Recording Scheme. We used chi-square test 

to analyse prevalence and t-test to analyse herd size differences. To 

analyse milk production we used a linear regression model 

correcting for breed and herd size. The seroprevalence for BRSV in 

the pooled samples was 83% in district A and 73% in district B. For 

BCV the prevalence was 61% and 55% in district A and B, 

respectively. Ten herds were negative for both BRSV and BCV. The 

prevalence in BTM was 95.1% for BRSV in district A and 92.1% in 

district B, and 100% for BCV in district A and 81.6% in district B. 

None of the herds were antibody negative to both viruses in BTM. 

There were no differences in herd size and milk production between 

positive and negative herds, both with respect to the pooled sample 

and the bulk tank milk. Negative herds were located in close 

proximity to positive, indicating that airborne transmission between 

herds is not of great importance for these infections and that it is 

possible to stay negative although virus is circulating in the area. 

Another interesting finding is that there were significantly (p

109 Association between Johne’s Disease Milk ELISA Test Result 

and Milk Production and Breed in Canadian Dairy Cows 

U. Sorge 1 , D. Kelton 1 , W. Sears 1 , J. Jansen 2 , A. Godkin 2 , R. Cantin 3 

1 

University of Guelph, Department of Population Medicine, Guelph, 

Canada 

2 

OMAFRA, Department of Population Medicine, Fergus, Canada 

3 

CanWest DHI, Guelph, Canada 

Johne’s Disease is often described as a production limiting disease. The 

objective of this study was to quantify the difference in milk production 

of JD milk ELISA positive cows and test negative cows, as well as to 

evaluate variations in test results due to breed. The test day information 

from 4,390 cows from 80 CanWest DHI herds in Ontario was available 

for this analysis. The cows were tested between March 2005 and March 

2006 with the AntelBio Johne’s Milk ELISA. A mixed model was 

fitted to investigate the association of a positive milk ELISA test with 

the estimated cumulative 305 day milk production. Only factors with a 

p-value 12 months of age) seropositive 

(ELISA) for MAP in a national whole herd serosurvey. Control herds 

were matched to case herds on the basis of the year of diagnosis of 

PTB in the case herds. Statistical analyses were performed using uniand 

multivariate conditional logistic regression with the herd as the 

experimental unit. Factors relating to disease history, herd size, 

neonatal feeding and management and grassland management were 

found to be significantly associated with herd PTB status (P

prevalence of JD within a herd. Of the herds 42.2% did not have any 

animals that tested positive, whereas 57.8% of the herds had at least one 

JD-test positive or suspect cow. Many of the farmers saw JD as a 

problem for their herd, yet believed that JD could be eradicated from 

farms. About two-thirds (69%) of the farmers suspected that JD entered 

the herd through a purchased animal, yet the first diagnosed JD case in 

52% of the herds was in a home raised animal. Responses to many of the 

RA questions were correlated; e.g., if one management area was 

considered unhygienic then other areas were also dirty. In this study, the 

JD prevalence was not associated with herd size, number of purchased 

animals in the last five years, number of farms purchased from, or the 

hygienic state of the calving pen (p > 0.1). Most of the farms had 

maternity box-stalls, which tended to be cleaner than non-box-stall 

environments. However, more than 50% of the farms did not remove 

calves from the calving pen within 30 minutes of birth. This extended 

exposure to a contaminated environment might explain why neither the 

age at removal from the dam, nor the colostrum source (e.g. artifical vs. 

from dams with unknown JD status), were associated with the apparent 

JD prevalence. In conclusion, the RA appears to identify critical areas 

correctly, although there are clearly areas of the instrument that need to 

be further refined. Given the homogeneous nature of responses to some 

of the questions, there is opportunity to reduce the length of the 

instrument while increasing its discriminatory power. 

149 The Effect of Infection with Mycobacterium Avium Subsp. 

Paratuberculosis on Tuberculin Testing in Suckler Herds in 


P. Mullowney, M. Good, E. Costello, D. Barrett 

Department of Agriculture, Fisheries and Food, Animal Health, 

Dublin, Ireland 

To assess the degree of interference with bovine tuberculin testing in 

suckler herds in Ireland that infection with Mycobacterium avium ssp 

paratuberculosis would be likely to cause. 

Aim: To assess the degree of interference with bovine tuberculin 

testing in suckler herds in Ireland that infection with Mycobacterium 

avium ssp paratuberculosis would be likely to cause. 

Methods: In the period 2002 to 2006 a total of 94 beef suckler herds 

submitted faecal samples to the Central Veterinary Research 

Laboratory, which yielded a culture positive result for Mycobacterium 

avium subspecies paratuberculosis (MAP). The bovine tuberculin 

testing history of these herds over a twenty-year period is analysed. 

These findings are compared with those from a control group of 243 

suckler herds which gave ELISA negative results as part of a survey 

which was conducted to estimate the prevalence of paratuberculosis 

(Johne’s disease) in the Irish cattle population. Tuberculin testing data 

on individual animals that were confirmed positive by faecal culture 

both prior to and subsequent to the sample submission were compared 

with those from matched animals in the control herds. 

Results: Thirty one percent of the animals in the infected group had 

had a response greater than 2mm to the avian tuberculin and 18% had 

had a response greater than 2mm to the bovine tuberculin. In the control 

group the responses were 24% to the avian tuberculin and 15% to the 

bovine tuberculin. Forty-four of the 94 case herds had been restricted 

for TB in the period 1989- 2007 (47%) whereas the figure for the 

control herds was 136 out of 243 (56 %). In the case herds 26% had 

lesions whereas in the control group 31% had lesions. 

Conclusions: These findings would suggest that in Ireland, which has 

a herd incidence of infection with Mycobacterium bovis of 5.87%, 

infection with Mycobacterium avium subsp. paratuberculosis may 

interfere with the diagnostic test used for M. bovis infection. 

Key words: M. bovis, M. avium subsp. paratuberculosis, bovine 

tuberculosis, Johnes 

193 Parameters of the Antioxidative Status in Calves and 

Growing Cattle 

T. Sattler, D. Haser, M. Fürll 

University of Leipzig, Medizinische Tierklinik, Leipzig, Germany 

Objective of study: The antioxidative status is an important system for an 

organism to fight free radicals. It consists of intracellular, extracellular, 

enzymatic and non enzymatic components. The development of those 

components in crowing animals is important for the ability to prevent 

diseases. Stohrer et al. (2004) measured a decrease of the serum 

antioxidants in calves in the first days of live, determined with the Trolox 

equivalent antioxidative capacity (TEAC). Other important parameters of 

the antioxidative status are the intracellular enzymes superoxide 

dismutase (SOD) and glutathione peroxidase (GPX). Objective of the 

study was to examine the development of parameters of the antioxidative 

status (SOD, GPX, TEAC) in young cattle.Material and methods: We 

examined 39 clinically healthy female Holstein Friesian calves and took 

blood samples on the first and 7 th day of life as well as on the end of the 

first, 3 rd , 6 th , 9 th , 12 th and 18 th month of life. SOD activity in erythrocyte 

lysate (Photochem ® , AnalyticJenaAG), TEAC concentration in blood 

serum (Spectrophotometer DU 640B, Beckmann) and GPX activity in 

heparinized whole blood (UV-method) were measured. The feeding was 

up to the 5 th day of life colostrums, up to the 3 rd month milk changer, hay 

and concentrates and after this gras silage, hay and concentrates. 

Results: The SOD activity in erythrocyte lysate increases in the first 

three month of life from 4967 U/g Hb (median, 1 st quartile 4495; 3rd 

quartile 5598) on the first day of life to 6966 U/g Hb (5661; 6827) on 

the third day of life and decreases later on to 4287 U/g Hb (3772; 

4931). The GPX activity in whole blood increased from 60,9 U/g Hb 

(49,5; 78,7) on the first day of life to 143,8 U/g Hb (120,6; 178) on the 

6 th month of life and stayed than on an even level. The TEAC 

concentration in serum showed also an increase from 254 µmol/l (220; 

286) on the first day of life to 307 µmol/l (258; 336) in the sixth month 

of life and after that a very slight but not significant decrease. 

Conclusion: In this study we measured a continuous increase of all 

measured parameters of the antioxidative status up to an age of six 

month. After this stayed the antioxidative status on an even level, 

except for the SOD activity which showed a decrease. The SOD and 

GPX activities as well as the TEAC concentrations in 18 month old 

cattle were on the same level as in grown up cows. The above 

mentioned decrease of TEAC in young calves was not to be found. 

Key words: calves, cattle, antioxidative status, SOD, GPX 

194 Acute Phase Proteins and Variables of Enzymatic and 

Hepatic Profile in Dairy Cows during the Pre- and Postpartal 

Period 

G. Kovac, Cs. Tothova, O. Nagy, H. Seidel, J. Novotny, J. Tomcova 



During the time around parturition, there are big changes in whole 

metabolism of dairy cows, which reflect multiple disturbances in the 

homeostatic control of the animals. The production of acute phase 

proteins by liver is one of numerous reactions, by which the 

organism responds to the impaired homeostasis. The objective of 

this study was to evaluate the concentrations of selected acute phase 

proteins: haptoglobin (Hp) and serum amyloid A (SAA) - in blood 

serum of dairy cows during the reproduction cycle in relation to 

some variables of enzymatic and hepatic profile - aspartate 

aminotransferase (AST), gamma glutamyl transpeptidase (GGT), 

alkalic phosphatase (AP), lactate dehydrogenase (LDH), creatine 

phosphokinase (CPK), and total bilirubin (TBil). The analyses were 

performed in 64 dairy cows of a Slovak spotted breed and its 

crossbreds. The animals were divided into groups according to the 

reproduction period from 4 weeks before parturition to 10 weeks 

after parturition. Hp and SAA were determined by method of 

enzyme linked immunosorbent assay using diagnostic kits in 

microplates. Significant differences we found in average values of 

the Hp and SAA concentrations in several groups during the 

monitored period (P

195 Changes in Plasma Beta-Carotene Levels during the 

Peripartum Period Relate to Ovulation of First Follicular 

Wave Postpartum in Dairy Cows 

K. Kida 1 , C. Kawashima 1 , F. Schweigert 3 , K. Sawada 4 , A. Miyamoto 2 

1 Obihiro University of Agriculture and Veterinary Medicine, Field 

Center of Animal Science and Agriculture, Obihiro, Hokkaido, 

Japan 

2 Obihiro University of Agriculture and Veterinary Medicine, 

Graduate School of Animal and Food Hygiene, Obihiro, Hokkaido, 

Japan 

3 University of Potsdam, Department of Physiology and 

Pathophysiology, Potsdam-Rehbruke, Germany 

4 DSM Nutrition Japan, Tokyo, Japan 

Introduction: The positive relationship between supplemental ßcarotene 

levels and reproductive function has been shown. However, 

relationship between physiological plasma‚ ß-carotene levels and 

reproductive function is unclear. In addition, the plasma‚ ß-carotene 

levels decrease from dry period to about first week (wk) postpartum 

(pp). This coincides with negative energy balance which affects the 

occurrence of first ovulation. The aim of our study was to investigate 

the changes in plasma‚ ß-carotene levels, metabolic status, milk yield 

and milk component during the peripartum period in ovular and 

anovular cows at the first follicular wave pp. 

Methods: Twenty-two multiparous Holstein cows were used for 

profiling plasma‚ ß-carotene and metabolites during the peripartum 

period. The cows were fed a TMR consisting of grass, corn silage and 

concentrate. Blood samples for‚ ß-carotene and progesterone analyses 

were collected from wk 3 prepartum to wk 3 pp. And samples for blood 

metabolites and milk component were collected 2 times during the 

experiment period. The ovulation of first follicular wave was confirmed 

by progesterone levels and ultrasound. The data were compared 

between the ovular and anovular groups. 

Results: Thirteen cows ovulated at the first follicular wave pp. Parity, 

length of dry period, calving interval, mastitis incidence and 305-days 

milk yield during the last lactation did not differ between two groups. 

Plasma‚ ß-carotene levels from wk 3 prepartum to wk 0 pp were lower 

in anovular cows than in ovular cows (P 5% of cells were inflammatory cells 

(predominately neutrophils some lymphocytes, and occasionally 

monocytes). To achieve 90% confidence in the within farm estimate 

approximately 20 cows were sampled per herd. 

Results: 30 farms and 547 cows were enrolled in the study. The farm 

size averaged 973 milking cows with a median of 881 cows (range 501 

- 2400). 229 cows (42%; exact binomial 95% CI: 38-46%) were 

positive for subclinical endometritis. Herd-level prevalence ranged 

from 7.6% to 80.0% of the sampled cows with 1 st quartile equal to 28% 

and the 3 rd quartile equal to 54%. 

Conclusions: The prevalence of subclinical endometritis in this sample 

population prior to first service is high and agrees with previous reports 

using smaller numbers of cows. There is a large amount of variability 

between herds. It appears there are as yet unidentified herd-level risk 

factors associated with either a high or low risk for endometritis 

between herds. 

198 A Sanitation Program for Swiss Dairy Herds with Udder 

Health Problems Caused by Intramammary Infections with 

Staphylococcus aureus 

M. Kirchhofer 1 , T. Kaufmann 1 , R. Eicher 2 , D. Strabel 3 , A. Steiner 1 

1 

University of Berne, Vetsuisse Faculty, University of Bern, Bern, 


2 

Private, Vetsuisse Faculty, University of Bern, Jegenstorf, 


3 

Rindergesundheitsdienst RGD, Lindau, Switzerland 

Objectives of the study: Staphylococcus aureus (S. aureus) as an 

agent of herd udder health problems is well known. In a study carried 

out by the Swiss Bovine Health Service between 1999 and 2004, S. 

aureus was identified as the responsible bacteria in 80% of the cases of 

herd udder health problems. It is also known that the diagnostic 

sensitivity of routine bacteriology of intramammary infections (IMI)

with S. aureus after single sampling does not exceed 75%. 

Furthermore, drug-based treatment of S. aureus IMI carries a moderate 

to poor prognosis. Therefore, a sanitation program (SP) for Swiss dairy 

herds with S. aureus udder health problems was developed. 

Materials and Methods: Cows of affected herds were allocated to 1 of 

3 groups. Group 1 consisted of cows with healthy udders (SCC < 

150.000/ml; no signs of clinical mastitis; no IMI with S. aureus). The 

second group consists of the suspected cows with a somatic cell count 

over 150.000 SCC/ml, with clinical mastitis or treated after a S. aureus 

positive milk sample. Cows of group 2 can be moved to group 1, if their 

milk samples have tested negative for S. aureus three times in a row at 

an interval of 14 days. Group 3 consists of all the cows with 

pathological diagnosis of the udder, with chronically high cell counts 

above 150.000 SCC/ml over several months or with IMI with S. 

aureus. The cows of this group are obliged to be culled or treated. The 

milking order of the cows during the SP matches systematically the 3 

sorted groups. According the SP, the udder health problem of a herd 

with 102 cows was solved. 

Results: This SP and its decision-making process can be presented as a 

quality assurance program. In dairy herds with IMI caused by S. aureus 

each cow will be tested 3 times. Infected cows have to be culled or 

treated and controlled. Six weeks after starting the SP, the group 1 with 

healthy cows can be established. The duration is based upon the 

development of the SCC of the herd. At the start of the SP, 52% of the 

lactating cows were infected by S. aureus. 19 months after starting, the 

bulk milk SCC decreased from over 500.000 under 100.000 SCC/ml. In 

this period, 108 of 524 milk samples tested positive for S. aureus and 

44 therapies with antibiotics were administered. The milk of all 

lactating cows tested negative. 

Conclusion: This experience shows that cleaning-up herds with an 

udder health problem due to S. aureus with this SP is possible under 

Swiss conditions. 

199 Characterization of Bovine Abortion Syndrome Incidence in 

Chilean Dairy Herds 

P. Gädicke, O. Alocilla, K. Amenabar, R. Becker, G. Monti 

Universidad Austral de Chile, Facultad Ciencias Veterinarias, 

Valdivia, Chile 

Objectives of study: The aim of this study was to estimate frequency 

and dynamics of bovine abortion syndrome and to identify groups of 

cows affected by abortion using quantitative epidemiological tools. 

Materials and methods: The study was performed using records of 

44,959 lactancies from 77 dairy herds coming from southern Chile, 

between years 2001 to 2005. An observed abortion case was defined as 

a case recorded in farmer’s reproductive records. An inferred abortion 

case was a non registered case inferred by the analysis of date’s 

differences of artificial insemination of cows within lactation. A 

general case of abortion was the occurrence of any of previous types. 

Incidence and incidence density (IDR) rates for first abortion was 

estimated, using lactation as unit of analysis. Possible associations with 

individual characteristics were inspected by logistic regression models. 

Results: From the total, 5,745 (11.6%) general abortion cases were 

found and 5,218 (90.8%) correspond to first abortion case. Out of them, 

929 (17.8%) were observed cases and 4,289 (82.2%) were inferred. 

Then, 487 lactations had 2 abortion cases during a lactation, where 366 

(75%) were inferred cases. Finally, 40 lactations had 3 abortion cases, 

where 29 (72.5%) were inferred cases. There is no statistical difference 

in the proportion of repeated abortion across parities (p>0.05).The IDR 

for first total abortion was 1.743% per cow-month. However, the 

estimated IDR for observed cases was 0.256% per cow-month. The 

second and third trimester of gestation had significant lower incidence 

rates (p

In the last year 2006, in the period between April and August the cattle 

industry subject to severe outbreaks of three epidemics all over Egypt. 

Foot and Mouth disease, Lumpy skin disease and Ephemeral fever were 

prevalent in this period and caused severe illness associated with acute 

clinical manifestations. Each disease expressed its own cardinal signs and 

in a drastic form. Epidemologically FMD spreed from time to time over 

the entire Egypt, which characterized by high morbidity and low mortality 

with expection in buffaloe farms in between young calves, which is 

considered as killing disease and characterized by sudden death without 

apparent clinical signs. The morbidity rate in cattle farms was ranged from 

25% to 60% and the mortality were ranged from 2.5% to 4%. Meanwhile 

in buffaloe calves the motility rate reached 25%. Lumpy skin disease was 

dramatically spreed in between the cattle either as herd population or 

sporadic cases. The most losses from lumpy skin disease were raised from 

the secondary complications; which caused severe losses. Ephemeral 

fever also has been recorded as severe illness affected cattle and Buffaloes 

and characterized by fever, lacremation, nasal discharge, off food and 

marked drop in milk yield, in addition to general emphysema, rumination 

is markedly decreased and associated with ruminal atony. Secondary 

complications were in the form of mastitis, bronchopneumia and 

recumbency. The overall stress and environmental factors which affected 

the wide spreed of the diseases were, high climatic temperature and 

humidity and increased population of insect vectors. Specific clinical 

pictures of each disease were illustrated in this manuscript. 

324 Bluetongue: Disease Awareness in Switzerland 

A. Tschuor 1 , H. Ochs 2 , C. Maret 2 , M. Hässig 1 

1 

University of Zurich, Department for Farm Animals, Zurich, 


2 

Federal Veterinary Office, Animal Health Division, Berne, 


As Bluetongue arrived in central Europe in summer 2006, veterinarians 

and farmers were very concerned that the disease could be diagnosed 

adequately when reaching Switzerland. Due to that, the Swiss federal 

veterinary office started a novel and unique campaign to increase the 

awareness for bluetongue in Swiss farmers in order to get them familiar 

with the clinical signs of the disease. It was supposed that by raising the 

awareness for the disease an eventual outbreak would be identified 

faster and earlier so that further spreading in Switzerland could 

hopefully be avoided. This campaign of the Swiss federal veterinary 

office included different strategies. Beside continuous distribution of 

news through the daily press an internet homepage 

(www.bluetongue.ch) was installed. This site contains daily actualized 

informations about the disease beside informations how to protect the 

animals from the disease etc. in different languages. Furthermore, 

different leaflets were distributed to the sheep and cattle farmers. The 

probably most effective tool for the disease awareness was the 

production of a DVD wherein the clinical symptoms were clearly 

demonstrated in different languages. This DVD was offered to the 

farmers for free and about 2500 copies have been distributed until 

today even outside the Swiss borders. Furthermore, a campaign was 

started wherein sheep farmers from all parts of Switzerland were 

elected to observe their animals for the disease. They are obligated to 

report in an interval all the diseases (and especially the symptoms) 

occurring on their farm to the federal office. This is also thought to be 

an effective early alarm system for bluetongue disease. All these 

measures will be evaluated by scientific analyses. The oral presentation 

of this Swiss disease awareness campaign at the WBC in Budapest 

should motivate other countries to use the different multimedia 

opportunities to inform farmers and the public about an upcoming 

disease and to show them up the efficiency by means of the DVD and 

the homepage. The different implications in producing such a DVD are 

outlined in the presentation. 

325 Serological Survey for Infectious Bovine Rhinotracheitis and 

Bovine Viral Diarrhea Viruses in Venezuelan Bovine Herds 

J. Alvarez 1 , A. Lozano 1 , R. Perez 2 , R. Paiva 2 , R. Guerrero 1 , 

J. Socorro 1 , A. Mazzei 3 , J. Rodriguez 1 , A. Colmenares 4 

1 

Pfizer Animal Health, Technical & Regulatory Dpt., Valencia, 

Venezuela 

2 

Veterinary Diagnosis Laboratory R. Paiva & R. Perez, 

Acarigua/Sta. Barbar, Venezuela 

3 

DiagVet, Valencia, Venezuela 

4 

CM Lab, Valencia, Venezuela 


The objectives of this survey were: 1- to get serological evidence of 

Infectious Bovine Rhinotracheitis (IBR) and Bovine Viral Diarrhea 

(BVD) in Venezuelan cattle herds; 2- to understand the dynamics of IBR 

and BVD in tropical cattle herds and, 3- identify when is the optimum 

time to apply vaccination programs against IBR and BVD in bovine 

herds. This serological survey was developed using commercial ELISA 

kit (Trachitest ® Serum Biphasic Kit by Bommelli and BVD Sero ® II Kit 

by Bommelli). From 2002 to 2006, 5,672 and 4,127 samples were 

processed by ELISA for IBR and BVD respectively. Samples were taken 

in non-vaccinated herds against IBR/BVD using a random schedule, 

considering all groups of ages (bulls, cows, heifers and calves) with a 

number of 40-100 samples by herd in 18 out of 23 geographical states of 

Venezuela. Age of animals tested was more than 4 months in order to 

avoid presence of maternal antibodies. All tests were performed by 

private veterinary laboratories. Results of this survey confirm 

previously reported data; IBR and BVD are highly present in 

Venezuelan bovine herds. All geographical states tested had at least one 

herd with at least one antibody-positive animal for IBR and BVD. 

Overall antibody-positive samples were 68.71% and 53.57% for IBR 

and BVD respectively. There were no differences between beef, dairy 

and double purpose herds for IBR and BVD. Marked differences 

between groups of ages were found; IBR and BVD are less prevalent in 

calves and heifers than cows and bulls. Samples antibody-positive in 

each age group for IBR were: calves 49.49%, heifers 73.45%, cows 

88.89% and bulls 80.77%. Results for BVD were: calves 35.71%, 

heifers 48.44%, cows 63.94% and bulls 37.84%. These findings suggest 

that venereal transmission between bulls, cows and heifers is very 

effective under natural breeding practices in the tropic, mainly for IBR; 

additionally, these results suggest that an early vaccination program 

against these viruses will help to build a strong immunological status in 

Venezuelan bovine herds to support field challenge. Finally, increased 

preventive actions should be taken in Venezuelan herds including 

vaccination schedules using IBR/BVD vaccines in order to improve 

reproductive performance and prevent shedding and dissemination of 

these viruses. The authors wish to thanks to all members of Pfizer 

Venezuela, as well as, all Venezuelan farmers, veterinarians and 

technicians whom supported and participated in this survey. 

326 Seroprevalence of Bovine Leukemia Virus Infection in Dairy 

Farms in Western Thailand 

T. Rukkwamsuk, S. Panneum 

Faculty of Veterinary Medicine, Kasetsart University, Large Animal 

and Wildlife Clinical Science, Nakhon-Pathom, Thailand 

Seroprevalence of bovine leukemia virus infection was studied in 50 

(26%) dairy farms from 192 farms belonging to Tha-muang Dairy Cooperatives 

in western part of Thailand. In this Co-operatives, the number 

of multiparous cows ranged from 4 to 64 (14 ± 10, ± s.d.) per farm, and 

the number of replacement heifers ranged from 1 to 36 (8-9) per farm. The 

ratio of cows and heifers was 0.65 to 1.00. In each selected farm, a 

maximum of 5 blood samples were collected from each group of 

multiparous cows or replacement heifers. Antibodies against bovine 

leukemia virus infection were determined using indirect ELISA (IDEXX 

HerdCheck Anti-BLV) in pooled sera from each group of animals. 

Positive results of pooled sera (from either cows or heifers) were rechecked 

for individual infection. Results revealed that herd 

seroprevalence was 60.0% (30/50). From 417 blood samples (246 from 

cows and 171 from heifers), seroprevalence was 41.1% (101/246) and 

19.3% (33/171) in multiparous cows and replacement heifers, 

respectively. These results indicated a high prevalence of bovine leukemia 

virus infection in dairy herds in western Thailand, which was observed in 

both cows and heifers. This epidemiological data is crucial for prevention 

and control measures of bovine leukemia virus infection in the future. 

Key words: bovine leukemia virus, dairy cow, prevalence 

327 The Effect of Enzootic Bovine Leukosis on Health, Milk 

Production and Fertility in a Dairy Herd: a Prospective Study 

A. Shoham 1 , M. Van Straten 1,2 

1 

Hachaklait, Caesarea Ind. Park, Israel 

2 

Hebrew University, Koret School of Veterinary Medicine, Rehovot, 

Israel 

Bovine leukemia virus (BLV) is a retrovirus that causes lymphosarcoma, 

persistent lymphocytosis and sub-clinical infections in cattle. Economic 

loss may be due to the clinical form of the disease, trade restrictions and

loss in milk production and reproductive performance. Results from 

studies investigating the relationships between BLV and health, milk 

production and reproductive performance have been controversial.The 

aim of this study was to investigate the effect of sub-clinical BLV 

infections on milk production, reproductive performance and survival in 

high producing dairy cows, while taking the point of time at which seroconversion 

occurred into account. A prospective cohort study was 

conducted in a commercial dairy herd consisting of 300 Israeli Holstein 

cows. From January 2006 until November 2007, serum samples were 

collected from all first lactation heifers (n=101) calving in 2006. 

Collection started 4 weeks before calving and continued every 3 months 

until the end of the study or until sero-conversion occurred. Samples were 

checked for antibodies against BLV using the agar gel immuno diffusion 

test. In total, 422 samples were collected. We used a discrete-time 

proportional odds model to investigate the association between the 

probability of undergoing sero-conversion and time or season. Health and 

reproduction data were analyzed using chi-square tests and Kaplan-Meier 

tests. Milk production and somatic cell counts were analyzed using 

marginal models for repeated measurements.By the end of the study, 59 

(58%) cows experienced sero-conversion. There was no difference in 

culling percentage (7%) between sero-positive and sero-negative cows. 

No particular risk period could be found for sero-conversion, nor was 

sero-conversion associated with season. No significant differences were 

found between the two groups in the proportion of cows suffering from 

post-partum diseases, unobserved heat, first insemination pregnancy rate 

and cumulative probability of conception. No effect of sero-conversion or 

of time of sero-conversion on milk production could be demonstrated.Our 

results indicate that the risk of BLV sero-conversion in an endemic herd is 

more or less constant in the 2 years following first calving and does not 

necessarily occur rapidly after introduction of heifers into the lactating 

herd. Within our study period, no negative associations between seropositivity 

for BLV and survival, health, reproductive performance, and 

milk production were found. 

328 Epidemiological Studies of Vampire Bats (Desmondus 

Rotundus) Attacks in a Brazilian Beef Cattle Herd 

E. Ortolani 1 , C. Maruta 2 


Department of Clinical Studies, Sao Paulo, Brazil 


The predation of cattle by vampire bats is quite common in Brazilian 

herds. The most important loss due to vampire bats is cattle rabies, but 

also low weight gain occurs by the bloodsucker habits of these animals. 

An epidemiological study was carried out throughout the year of 2007 in 

a beef cattle herd raised extensively in the State of Sao Paulo, Brazil. In 

order to know the distribution of the attacks throughout the year the 

number of recent bitten cattle was counted every fortnightly in the herd 

that varied from 187 to 205 animals during the year. On day July 2 nd it was 

counted the number of cattle (total 205) with recent bites to evaluate the 

influence of the category, sex, breed of the animal, the number of 

bites/head, the anatomical region attacked on the vampire bat predation. 

The complications of the bites in the wounds were also recorded. The herd 

was vaccinated against rabies twice a year. The percentage of cattle 

recently bitten throughout the months of the year was similar. The attacks 

were higher (53%) on weaned calves (5 m to 1 y old) than cows (11.8 %), 

steers & bulls (19.5%), and heifers (31.2 %) (P < 0.03); cows were less 

bitten than lactating calves (33.3%) (P < 0.04). There was no distinction 

between sex (P > 0.8). The Simmental cattle (52.9 %) were much more 

bitten than Zebu (14.1%) and Charolais (23.7%) crossbred cattle. Most of 

the cattle had a single recent bite (53 %) followed by two (23.5 %) or more 

than two (3 up to 6) bites (23.5 %). The most frequently anatomical area 

attacked was the neck (60.2 %), followed by the tail (21.5 %), dorsal (9.7 

%), head and ears (4.3%), members (3.2%) and shoulders (1.1%). Three 

weaned calves had to be treated during the year against myiasis and two 

for abscesses that developed in the wounds. 

Key words: epidemiology, vampire bats, bites, breeds, cattle 

329 Difference in Virus Excretion and Transmission between 

Calves and Dairy Cows in the Course of an Acute FMDV 

Infection 

K. Orsel 1 , A. Dekker 2 , A. Bouma 1 , J. Stegeman 1 , M. De Jong 3 

1 

Utrecht University, Farm Animal Health, Utrecht, The Netherlands 

2 

Central Institute for Animal Disease Control CIDC, Lelystad, The 


3 Wageningen University and Research Centre, Quantitative 

Veterinary Epidemiology, Wageningen, The Netherlands 

Objective: The aim of this study was to determine differences in virus 

excretion and virus transmission between calves and dairy cows after 

infection with FMDV. 

Material and Methods: In two experiments, one with groupd of calves 

of 10 weeks old and one with lactating dairy cows. Half of each group 

was inoculated with FMDV isolate ONET/2001 and the other half of 

each group was exposed by direct contact to these animals. Clinical 

inspection, virus titration and serological tests were performed. To 

quantify virus transmission the reproduction ratio R was estimated with 

a maximum likelihood estimate using an S-I-R model. 

Results: In the calves mild clinical signs were observed and not all 

contact exposed calves became infected. Dairy cows showed severe 

clinical signs and all in-contact animals became infected. The total 

amount of excreted virus differed significantly between inoculated and 

contact-infected animals and also between infected calves and cows. 

Inoculated cattle shed virus for a longer time compared to contact 

exposed cattle. The transmission ratio for calves Rcalves was estimated 

2.52 [1.13;52.1], and for dairy cows R cows= Inf [1.3; Inf]. Both R 

values were significantly above 1 (p

serious economic concern in the fattening units of young bulls in 

western France, even though vaccination and antibiotic prevention are 

widely implemented. 

Key words: young bulls, respiratory disorders, growth, economic 

impact 

360 Pathogen Exposure and Incidence of Respiratory Disorders in 

Young Bulls 

S. Assié 1 , H. Seegers 1 , G. Denis 2 , N. Bareille 1 

1 Veterinary School & INRA, Animal Health Division, Nantes, France 

2 Cavac, La Roche sur Yon, France 

Respiratory disorders caused by different pathogens affect frequently 

young bulls in fattening units. The objectives of the study were to 

describe the incidence of respiratory disorders during the first 40 days on 

feed and the exposure of individual young beef bulls and of batches to the 

Bovine Respiratory Syncytial Virus (BRSV), Parainfluenza-3 Virus 

(PI3), Bovine Viral Diarrhoea Virus (BVD), Bovine Herpesvirus type 1 

(BHV1) and to the bacteria Mycoplasma bovis (M. bovis) and 

Mannheimia haemolytica (M. h.). The study was performed between 

September 2005 and June 2007 in western France. Seventy-one farms 

were included. The animals were reared in batches of 11 animals on 

average. They belonged to 100 batches of which 90 pens were composed 

only of purchased young bulls. Blood samples collected at the first day 

on feed and 6 weeks later were tested. Antibodies specific to the viruses 

and M. bovis were measured by ELISA tests for routine diagnostic (in the 

LDA 85, La Roche sur Yon, France). The testing for M. h. specific 

antibodies was performed in the laboratory of Intervet (Boxmeer, The 

The Netherlands) using an in-house ELISA test. Mortality rate was 0.5 % 

and incidence rate was 17.4 % during the first 40 days on feed. Cases 

occurred in 53 batches out of 100. In these 53 batches, the number of 

cases per young bull on feed was 0.25 and varied from 0.05 to 0.92, with 

a median value of 0.25. The percentage of young bulls seropositive at the 

first day on feed was very high for M. h. (80%) and for the PI3 (57%). 

The percentages of young bulls that seroconverted against VRSB, PI3, 

M. bovis and M. h., respectively 80, 79, 72, 61, were high. All 68 batches 

were exposed with at least 2 pathogenic agents prior to the beginning of 

the fattening. Almost all animals in a batch were concomitantly exposed 

to several pathogens. For all investigated pathogens, the percentage of 

diseased bulls was higher in the groups with a high seroconversion rate, 

For BRSV, BVD and M. h. the percentage of diseased bulls was lower in 

the groups with a higher percentage of animals that were seropositive at 

the first day on feed. The incidence rate in the first 40 on feed was quite 

high. Simultaneous exposure of groups of young bulls to the viruses 

BRSV, PI3 and BVDV as well as for the bacteria M. bovis and M. h. was 

frequent. Further studies on the etiology of respiratory disorders should 

take this simultaneous exposure into consideration to allow reliable 

conclusions. 

Key words: young bulls, respiratory disorders, incidence, pathogens 

361 Bovine Theileriosis by Theileria buffeli in Southern Italy: 

Epidemiology, Clinical Case and Exsperimental Infection 

L. Ceci, P. Paradies, D. De Caprariis, M. Sasanelli, F. Iarussi, 

G. Carelli 

University of Bari, Animal Health and Welfare, Valenzano (Bari), 

Italy 

Objectives of study: The aim of the present work is to molecularly 

evaluate the presence of Theileria species in cattle of southern Italy and 

to investigate the possible pathogenic role of Theileria buffeli. A 

clinical case of theileriosis by T. buffeli in a dairy cow is described and 

the results of an experimental infection in a splenectomised calf are 

reported. 

Materials and Methods: A clinical-epidemiological survey on bovine 

tick borne diseases (TBDs) has been conducted in southern Italy. 150 

dairy farms have been randomly selected and in each one 10 adult cattle 

have been clinically evaluated and bleed. Reverse line blotting 

hybridization (RLB) was performed on all samples using 

oligonucleotides for the group Ehrlichia/Anaplasma and 

Theileria/Babesia and specific ologonucleotids for single pathogens 

including Theileria annulata, Theileria mutans and T. buffeli. During 

the study a clinical case was observed in a 3 years old Holsten Friesan 

cow showing fever (41 °C), depression, weight loss, hypogalaxia and 

mild lymph-nodes enlargment. Furthermore a splenectomized calf was 

experimentally infected using whole blood collected from a cow with 


symptoms of TBDs and showing 17% parasitemia for Theileria spp. 

and < 1% for A. marginale. The animal was daily monitored by means 

of clinical examinations and laboratory exams. 

Results: Out of 1500 tested samples 409 (27.3%) resulted positive for T. 

buffeli at RLB. No other Theileria species were detected. Theileria spp. 

(with a parasitemia reaching 30% was revealed at blood smears 

microscopical examination in the cow showing clinical symptoms and at 

RLB a single infection by T. buffeli was identified. The cow was orally 

treated with primaquine phoshate for three days.The calf experimentally 

infected developed mild fever and anemia. A. marginale was initially 

detected on blood smears and the calf was successfully treated with 

oxitetracicline. In a second time the calf was immunosuppressed with 

desametasone; in four days the animal developed again mild fever and 

anemia. At this time the blood smears microscopical examination revealed 

the presence of Theileria spp. with a parasitemia reaching a peak of 5%. 

Conclusions: T. buffeli is the only Theileria specie revealed in the 

investigated area of Italy. Furthermore, although it has been considered 

a non pathogenic parasite of cattle our data suggests its possible 

pathogenic role in immunodeficiences and/or coinfection conditions. 

Key words: bovine theileriosis, Theileria buffeli, clinical case, 

experimental infection 

362 Study on Leptospira Borgpertesenii Serovar Hardjo Type 

Hardjo Bovis in Cattle and Aborted Fetuses from Venezuela 

Monitored by cELISA vs. MAT 

E. Angulo 1 , V. Bermúdez 2 , J. Alvarez 3 , J. Rodríguez 3 , 

A. Moscardi 4 , O. Verde 2 , R. Pérez 5 , A. Morales 2 

1 Universidad de Carabobo, Patologia, Valencia, Venezuela 

2 Facultad de Ciencias Veterinarias UCV, Patologia Veterinaria, 

Maracay, Venezuela 

3 Pfizer Animal Health, Technical & Regulatory Dpt., Valencia, 

Venezuela 

4 Ladivet, Diagnosis Laboratory, Maracay, Venezuela 

5 Veterinary Diagnosis Laboratory R. Paiva & R. Perez, Diagnosis 

Laboratory, Acarigua/Sta. Barbar, Venezuela 

Leptospira borgpetersenii hardjo type hardjo bovis (LHB) is the most 

common cause of abortion in beef and dairy cattle in USA (Bolin, 

2001), while L. interrogans hardjo type hardjo prajitno (LHP) is more 

prevalent in UK and Europe. The objective was to determine the 

seroprevalence of LHB in cattle and aborted fetuses (2007) from 10 

states of high incidence of leptospirosis in Venezuela. To determine 

presence of antibodies (Ab) against LHB in cattle, 42 farms were 

included from 10 strategic states across the country (n = 1,743). A 

random sampling schedule of 20 cows, 15 heifers and 5 bulls per farm 

was developed. A competitive ELISA (Linnodee ® , UK) was 

standardized, run and read at 460 µm filter (Automated system Elisa 

Biotek ® ) as recommended by manufacturer. Positive animals on 

cELISA for LHB were tested by MAT (using a routine strain of LHP, 

supplied by INH Rafael Rangel). The degree of agreement between 

LHB versus LHP on cELISA and MAT was evaluated y chi-X 2 and 

ANOVA. Confounders were controlled statistically by Mantel & Hansel 

technique, by using SAS package of UCV. To assess the Abs to LHB in 

bovine fetuses, fetal body fluids pool of 80 aborted Fetuses and sera 

from their respective dams were evaluated and compared to 29 

immunocompetent fetuses (>5m) obtained from slaughterhouses. 

Positives samples on cELISA were similarly studied and compared by 

MAT as done above. The overall seroprevalence was 61%, but in cows 

was higher (72.4%) than heifers (58.6%) and bulls (56.5). All sampled 

states (n = 10) showed a high seroprevalence to LHB being the lowest 

32% and highest 79%. Positive cELISA sera Ab to LHB (89%) and in 

fetal fluids (58%) were much higher than those detected by MAT (43% 

& 12%). Fetuses and dams showed cELISA Abs to LHB in 45% and 

20% respectively. Fetuses collected at slaughterhouses did not reveal 

Abs on cELISA nor MAT on fetal fluids. These results seemed to 

express that a higher number of calvings a major venereal exposure to 

Leptospira occurs. Despite the fact the sampled bulls at each farm was 

small the elevated prevalence may support the role they play in 

venereal exchange and infection rate by this route in leptospirosis. 

Despite the fact that this test use a mAb to LHB it can cross react with 

LHP antigenic surface. But the fact that those positive animals and 

fetuses on cELISA were tested by MAT, whose strain was solely LHP, 

expressed a greater level of Ab concentration, 58% over the MAT test. 

The authors wish to thanks to all members of Pfizer Venezuela & 

Caribbean Area, as well as, all Venezuelan farmers, veterinarians and 

technicians whom supported and participated in this survey.

363 Modified Live Virus Bovine Viral Diarrhea Virus (BVDV) 

Vaccination of BVDV Maternal Antibody Positive Calves 

Followed by BVDV Challenge 7 Months Later 

A. Zimmerman 1 , R. Buterbaugh 1 , J. Schnackel 2 , C. Chase 1 

1 

Rural Technologies Inc., Research Project, Brookings, United States 

2 

Fort Dodge Animal Health, Livestock Technical Services, Fort 

Collins, United States 

Objective: This trial was conducted to determine if young calves that 

had consumed colostrum containing Bovine Viral Diarrhea Virus 

(BVDV) antibody could be immunized with an adjuvanted Modified 

Live Virus (MLV) BVDV vaccine. 

Materials and Methods: Twenty-two neonatal calves obtained prior 

to consuming colostrum were randomly divided into three 

groups. Group 1 (N=6) were negative controls and were raised 

colostrum deprived. Within 24 hours of birth, calves in Groups 2 and 3 

were given pooled colostrum obtained from cows vaccinated 6 to 8 

weeks prior to calving with an inactivated BVDV Type 1 and 2 

vaccine. Group 2 calves (N=8) were vaccinated once between 4 and 5 

weeks of age with a MLV BVDV Type 1 and Type 2 vaccine. Group 3 

calves (N=8) served as maternal antibody positive controls and were 

not vaccinated. Serum neutralizing antibody for BVDV was 

determined on all calves at intervals of 5 weeks or less. When all calves 

in Group 3 became seronegative (8 months of age when challenged with BVDV. Mortality 

in Group 1 was 33% (2/6) (p =0.08) compared to 0% (0/8) in Group 2 

calves. Four of 8 (50%) calves in Group 3 died which was significantly 

less than Group 2 calves (p = 0.02). From -1 DPC, calves in Group 1 

lost 28.5 kg and 9.8 kg on 14 and 21 DPC, respectively. Calves in 

Group 2 gained 1.6 and 9.7 kg by 14 and 21 DPC. Calves in Group 3 

lost 25.2 and 25 kg on 14 and 21 DPC. By 9 DPC, combined clinical 

scores were less in Group 2 calves compared to Groups 1 and 3. Group 

2 calves viremia was 0% on days 8 and 10 through 21 post challenge; 

Group 1 calves reached 83% viremia on 4 and 6 through 8 DPC and 

declined to 0% on day 13 post challenge. Group 3 calves had 100% 

viremia 7 and 9 DPC decreasing to 20% by 13 DPC. 

Conclusion: Young calves developed some protective immunity from 

MLV BVDV vaccination despite having significant circulating BVDV 

maternal antibody at the time of vaccination. 

Key words: Bovine Virus Diarrhea Virus, BVDV 

364 Survival Data of 810 Dairy Cows Corrected for Left 

Displaced Abomasum (LDA) 

K. Sterner 1 , J. Grymer 2 , P. Bartlett 3 

1 

Sterner Veterinary Clinic, PC, Veterinarian, Ionia, United States 

2 

Galten Dyreklinik, Veterinarian, Galten, Denmark 

3 

Michigan State University, Epidemiology, East Lansing, United 

States 

The objective of this study was to determine which factors influenced 

the culling or death of cows following left displaced abomasum (LDA) 

repair, using either the Grymer/Sterner roll and toggle method or 

laparotomy. In this cohort study, information regarding the method of 

LDA repair and risk factors for LDA survival was collected for one 

year, and outcomes compared at 14 and 60 days post LDA repair 

among three groups: veterinarians performing roll and toggle repair (V- 

R&T), herdsmen performing roll and toggle repair (H-R&T), and 

veterinarians performing surgical repair (V-Surg.). Survivorship at 14 

days post LDA repair for the V-R&T group was 87% (286/329), 81% 

(327/403) in the H-R&T group, and 85% (66/78) in the V-Surg. 

group. Survivorship results at 60 days were: V-R&T 79% (260/329), 

H-R&T group 71% (286/403), and V-Surg. group 73% 

(57/78). Multivariable analysis indicated that current mastitis status, 

previous LDA, high pre-operative risk, and correction by herdsman 

rather than by veterinarian, were positively associated with failure to 

survive in the herd 60 days post LDA repair. The clinical relevance of 

this study confirms that correction of LDA by veterinarians using the 

Grymer/Sterner method of repair produces survival results that are 

generally comparable to open surgical correction of LDA. While 

survivorship rates at days 14 and 60 differed significantly between the 

V-R&T and H-R&T groups, bivariable analysis indicated that trained 

herdsmen could use the R&T procedure to repair LDA cows at 

survivorship rates within the range experienced by practicing 

veterinarians. 


1080 Studies on Phenotypic and Genotypic Features of 

Staphylococcus aureus Isolated from Human and Animals 

N. Ünal, E. Istanbulluoglu 

Faculty of Veterinary Medicine, Microbiology, Kirikkale, Turkey 

The aim of this study is to determine susceptibility patterns of several 

antibiotics to S. aureus strains isolated (46 from bovine milk samples 

with mastitis, 35 from bovine teat skins, 3 from bovine noses, 3 from 

caretaker hands and 3 from caretaker noses) from dairy farms in 

Kirikkale province by E-tets and define genotypic characteristic of 

these isolates by plasmid and Pulsed Field Gel Electrophoresis (PFGE) 

analysis. S. aureus isolates resistant to penicillin G, tetracycline, 

erythromycin, oxacilline, enrofloxacin were found to be 85.4 % (82), 

39.6 % (38), 5.2 % (5), 3.1 % (3) and 1.0 % (1), respectively. Nine 

different types of plasmid profiles were determined in the isolates by 

plasmid analysis. In 87 (90.6 %) of the isolates, 10 different types of 

plasmids having the size between 1.8 -19 kb were determined while no 

plasmid was detected from the 9 (9.4 %) of the isolates. S. aureus 

isolates were performed using PFGE method, 42 distinct PFGE 

patterns were identified. Strains were assigned as 13 major lineage 

groups (A, B, C, D, E, F, G, H, I, J, K, L, M) with respected to the 

genetic relationships. The percentage of PFGE pulsotypes of strains 

were clustered as 58.3 % (56 samples) A. 16.7 % (16) B, 5.2 % (5) C, 

5.2 % (5) G, 3.1 % (3) D, 2.1 % (2) E, 2.1 % (2) L, 2.1 % (2) M, 1.0 % 

(1) F, 1.0 % (1) H, 1.0 % (1) I, 1.0 % (1) J and 1.0 % (1) K, respectively. 

In conclusion; according to plasmid analysis and PFGE data, a limited 

number of S. aureus clones was detected in dairy farms in Kirikkale 

province. 

Key words: antibiotic resistance, MIC, plasmid analysis, PFGE, S. 

aureus 

1081 Effect of Grazing Management on the Prevalence of Tick 

Infestation and Exposure to Theileria parva (East Coast 

Fever) in Cattle of Smallholder Dairies in the Lilongwe Milk 

Shed Area 

J. Musser 1 , M. Ng'oma 2 , K. Russell 1 , M. Mulumba 3 , M. Mfitilodze 2 

1 Texas A&M University, Veterinary Pathobiology, College Station, 

United States 

2 Bunda College, University of Malawi, Animal Science, Lilongwe, 

Malawi 

3 Centre for Tick and Tick-Borne Diseases, Lilongwe, Malawi 

East Coast Fever (ECF), caused by Theileria parva, causes major 

economic losses in the dairy industry in Malawi. A study was 

conducted to determine the effect of grazing management schemes on 

tick infestation and the prevalence and incidence of T. parva in dairy 

cattle within the Lilongwe milk shed. The study was carried out 

between Aug 2005 and Apr 2006, with four sampling periods to 

incorporate different climatic seasons of dry, end of dry, wet, and end 

of wet seasons. The grazing systems were stratified into zero-grazing 

with cut and carry of grasses (animals=51; farms= 21) or open-grazing 

on natural pasture (animals=21; farm= 6). Head tick counts were 

conducted, and blood was collected for serology. Using an indirect 

fluorescent antibody test, a titer of 1:40 was considered positive for 

exposure to T. parva. At two sampling periods, wet season and end of 

wet season, significant differences were observed in the number of 

cattle infested with ticks (p

was a cumulative incidence rate of 27% and 32%, respectively, for 

exposure to T. parva during the study period. This study indicated that 

grazing management influences the number of tick-infested animals 

and the total number of ticks/animal. It also indicated that new 

infections to T. parva were taking place mainly during the wet season. 

The prevalence and incidence of animal’s sero-positive to T. parva may 

not be affected by grazing management or season, except for an 

increase in the incidence in zero-grazing animals during the wet season. 

A zero-grazing system may expose animals to fewer ticks, but animals 

are still at risk of infection to East Coast Fever. 

Key words: theileria, dairy, East Coast Fever 

1082 Elisa Antibodies to Study the Spread of Mycoplasma bovis 

in 2 Belgian Herds 

D. Gevaert 1 , P. Passchyn 2 , H. Theys 3 

1 Bayer SA-NV, Animal Health Division, Brussels, Belgium 

2 DAP De Toren, Veterinary Medicine, Torhout, Belgium 

3 DAP Theys, Veterinary Medicine, Turnhout, Belgium 

Objectives of the study In 2 herds suffering from serious bovine 

respiratory disease (BRD) initiated by Mycoplasma bovis, serology 

was used to study the spread of the bacterium. 

Materials and Methods Herd 1: a dairy herd of 117 Holstein Friesian 

cattle suffered since 2001 from BRD: relapses and growth retardation 

(calves) and chronic coughing (dairy cows). Tilmicosin and 

tulathromycin treated calves relapsed after 14 days. Calves are housed 

individually up to 2 weeks, followed by group housing in 3 age groups (1, 

2 and 3 months) in the same stable. The calves are vaccinated against IBR, 

Pasteurella haemolytica, Parainfluenza and BRSV. A blood sample of all 

cattle was analyzed for M. bovis Elisa antibodies (Bio-X, Belgium). Herd 

2 (open veal calf operation, 5,768 calves). Newly bought-in calves aged 2 

weeks are housed individually and fed milk replacer up to 6 weeks, 

followed by group housing and feeding roughage. A compartment 

consists of 50 to 58 calves. The BRD problems start 10-14 days after 

arrival, some calves developing arthritis 2 weeks later. Seriously affected 

calves face considerable growth retardation. Mortality between arrival 

and slaughter is 5%. Paired blood samples (upon arrival and 1 month 

later) and blood samples of 5 calves aged 2 up to 8 weeks were analyzed 

for M. bovis antibodies. 

Results Herd 1: the M. bovis antibody prevalence increased with age: 

25% in calves

1085 Vertical Transmission of Enzootic Bovine Leucosis Infection 

in a Dairy Herd with High Prevalence of the Disease in 

Uruguay 

R. Sienra 1 , H. Guarino 2 , G. Tort 3 , A. Hernández 3 , F. Olariaga 3 , 

J. Bianchi 3 , J. Slavica 3 , M. Alzugary 3 

1 

College of Veterinary Medicine, Large Animal Clinic, Montevideo, 

Uruguay 

2 

College of Veterinary Medicine, Microbiology, Montevideo, 

Uruguay 

3 

Assistant, Research project, Montevideo, Uruguay 

Enzootic Bovine Leucosis (EBL) in Uruguay is widely distributed among 

dairy cattle in the country The objective of this research was to determine 

the importance of vertical transmission of the LBE in one commercial 

dairy farm with a high prevalence of EBL infection, with a population of 

513 Holstein cows. During the calving period a permanent control of births 

were performed during the 24 hours, with the purpose of obtaining blood 

samples from the newborn calves prior to the ingestion of colostrum. After 

birth calves remained with their mothers for 6 to 8 hours of life, and then 

they were removed to the breeding area and feeding individually with pool 

colostrum and milk. A second blood sample was obtained at 24 hours postpartum. 

Serum samples of mothers and calves were obtained by puncture 

jugular without anticoagulant. The sera were processed through a 

commercial kit EBL, indirect ELISA technique to detect the gp-51 surface 

glycoprotein according with the indications of the manufacturer (IDDEX, 

USA). The prevalence of infection in the herd was 60,4%, with 310 

positive cows out of a total of 513. For the study of vertical transmission 99 

deliveries were included, 52 from seropositive mothers and 47 from 

seronegative cows. Before the intake of colostrum, all of the calves born 

from uninfected cows had similar status as their mothers (47 in 47). In the 

other hand, 9 of the 52 calves born from seropositive cows tested positive 

to EBL before the ingestion of colostrum (17%) In the second sample, 23 

of the calves whose mothers were negative were detected with EBL 

antibodies (seroconvertion rate of 50%). For the group of calves born from 

infected cows, at 24 hours of live, 34 tested positive (65%).The 34 calves 

included 9 detected infected at birth and 25 new positive animals 

(seroconversion 58%).The rate of vertical transmission observed in this 

study was higher than the reported of several authors, which found 

percentages between 6-8 %. The seroconversion during the first 24 hours 

of life means that in practice it is very difficult to identify infections vertical 

face of the presence of antibodies acquired after birth due to the 

administration of colostrum and milk mixture of different cow’s 

serological condition. In practical situations calves can not be controlled 

before the ingestion of colostrum. Therefore EBL positive status calf 

bleeding during their first days of life must be interpreted with caution. 

Proyect Financed by PDT-CONICYT, Uruguay 

Key words: Bovine Leukosis, vertical transmission 

1086 Abscesses Disease in Small Ruminants of Batna Area 

(Algeria): Preliminary Study of an Epidemiologic 

Investigation 

M. Alloui 1 , A. Ayachi 1 , J. Kaba 2 

1 University of Batna, Veterinary Science, Batna, Algeria, Algeria 

2 Agricurtural University of Warsaw, Veterinary Faculty, Department 

of Epizootiology, Warsaw, Poland 

The results of epidemiologic and clinical investigations, relating to the 

abscess disease at the small ruminants (goat and the sheep) in the area of 

Batna (Algeria) are presented. The epidemiologic study has concerned 18 

extensive flocks into extensive whose the number varied between 39 and 

102 heads of animals. The clinical studies were carried on 68 animals 

resulting from these flocks. Clinically, the disease didn’t affect the general 

state of the animal and often touched various parts of the body by one or 

two abscesses per animal affected. The internal form of the disease was 

not observed on 38 carcasses slaughtered in the slaughter-house of the 

area. The morbidity of the disease was 1.5 p. 100 in the goat and of 5.5 p 

100 in the sheep. This disease has often touched the adults whose age was 

higher than one year. The frequency of the abscesses of the head (parotid 

and mandibular, and retropharyngeal region) was higher than the other 

parts of the body (forelimbs and posterior) (68 p. 100 against 32 p. 100) at 

the two animal species. The supporting factors of the disease appearances 

were: the breeding of several animal species in the same farm (sheep, 

goat, bovine, and poultries), bad hygiene of the shelters, mode of course 

(mountains, meadows) and starvation (food shortage). 

Key words: abscess disease, epidemiology, small ruminants, Algeria 

1087 The Case of BVDV-MD in a Heifer 

A. Sagkan-Ozturk, A. Coskun, I. Sen 

University of Selcuk, Faculty of Vet. Med., Department of Internal 

Medicine, Konya, Turkey 

Diseases associated with Bovine Viral Diarrhea Virus (BVDV) have been 

recorded in most countries where cattle are raised and in some countries 

may be the single most important virus infection of cattle. BVDV is a 

pestivirus responsible for considerable economic losses in cattle industry. 

There are many BVDV-infected cattle without typical clinical 

manifestations of diarrhea and mucosal diseses. A large proportion of 

BVDV infections are also subclinical and majority seropositive cattle. 

Objective of study: The aim of this case report is to indicate BVD-MD 

-infected a heifer with the severe diarrhoea and high fever along with 

oral lesions. 

Materials and Methods: Material of the case report was constituted 

BVDV-MD -infected a Holstein- heifer, 16 months of aging. After 

clinical examination, venous blood sample was taken from vena 

jugularis of heifer. Taken blood samples were tested for BVDV antigen 

and neutralising antibody titres. A commercial direct and indirect 

ELISA kits (Institute POURQUIER) were used for the determination 

of BVDV antigen and neutralising antibody titres. 

Results: In routine clinical examination of heifer were observed small 

lesions in palate and salivation along with high temperature. There was 

diarrhea with bloodly and mucus in infected-heifer. BVDV antigen in 

blood samples of heifer was detected as positive. But the presence of 

BVDV antibodies in serum was seronegative. 

Conclusion: Cattle that clinical symptoms such as oral lesions, high 

temperature and diarrhoe with bloodly and mucus might be test for 

BVDV. 

This study was supported by University of Selcuk, Scientific Research 

Project Coordinating 

Key words: BVDV-MD, heifer, ELISA. 

1088 Statistical Correlation on Serological Surveys of Bluetongue 

Disease with Climate Changes in Northwest of Iran 

S. Mahdavi 1 , A. Bahonar 2 , AG. Ramin 3 

1 Razi Institute, Virology, Karaj, Tehran, Iran 

2 University Of Tehran, Veterinary Medicine, Tehran, Iran 

3 Urmia University, Animal Science Department, Urmia, Iran 

In the present study, the seroprevalence of bluetongue disease (BTD) was 

determined in 981 sera samples and their interrelationship to biological 

vector and climatic conditions as temperature, speed of windflaw, sunny 

hours, rainfall and humidity. The sera were collected from western 

Azerbaijan province of Iran in 2003. The prevalence of BTD was 

evaluated for serodiagnosis purposes (without considering of age, species 

and sex), based on an Agar Gel Immunodiffusion test (AGID). The 

infection rate was found to be 55.9%. The proportion of positive sera in 

different months were significant (P

The level of antibodies produced in colostrum following injection of 

the trivalent Rota-corona virus and E. coli vaccine was evaluated to 

understand the prophylaxic effect of vaccine against calf diarrhea. The 

number of 46 pregnant cows from industrial dairy herd were selected. 

A 5 mls trivalent Rota-corona virus and E. coli vaccine was injected 

subcutaneousely in 7 months pregnant cows. After parturition a 50 mls 

colostrums was collected and evaluated for antibodies using 

compatative ELISA method and commercial kit of Rota-corona virus 

and E. coli (Bio X, Belgium). Meanwhile, 27 pregnant cows which 

have not vaccinated were used as control group. From out of 46 

vaccinated cows, antibodies against E. coli was detected in 33 cases 

(71.7%) including one positive were 4 cases (8.7%), two positive were 

28 cases (60.9%) and four positive was one case (3.1%). In control 

group 23 cases were negative (85.2%) and one positive were 3 cases 

(11.1%) and two positive was one case (3.7%). From out of 46 

vaccinated cows, antibodies against rota virus were detected in all 

samples (100%) including one positive were 3 cases (6.5%), two 

positive were 43 cases (93.5%). For corona virus one positive were 9 

cases (19.5%) two positive were 34 cases (73.9%) and 3 positive were 

3 cases (6.5%). All control group for rota and corona antibodies were 

two positive. The highest and lowest inhibition percent in vaccinated 

cows for E. coli antibody were 85% and 1%, rotavirus were 59% and 

20% and corona virus were 61% and 20%, respectively. Chi-Square 

test shows significant difference (P

Jörgensen-Kutas method. Physiological range for rumen pH was taken 

between 6.2 and 7.2; pH below 6.0 was considered subclinical rumen 

acidosis and above 7.5 subclinical rumen alkalosis. Physiological ranges 

for urine parameters were between 7.0 - 8.4 for urine pH, 2.5 - 4.8 for 

urine base-acid ratio and 80 - 220 mmol/L for NABE. 

Results: Rumen acidosis was found in 43 (12.3%), rumen alkalosis in 

30 (8.6%) of the 348 samples; the pH of the remaining 275 (79.1%) 

samples were within the physiological range. Rumen pH showed 

significant but week correlation coefficients to NABE (rs = 0.13), urine 

base/acid ratio (rs = 0.13), and to urine pH (rs = 0.19). Overall, the 

sensitivities of the urine parameters to predict rumen acidosis (NABE 

44.8%, b/a-ratio 55.8%, urine pH 2.3%) were moderate or low. 

Specificities for rumen acidosis of the parameters were 64.5% for 

NABE 47.9% for base/acid ratio, and 97.0 % for urine pH. 

Conclusions: We conclude that the measured parameters of urine acid 

base status showed a moderate or even week association to rumen pH. 

The diagnostic value of these parameters seems to be limited to assess 

rumen acid-base status in healthy dairy cows during transit period and 

early lactation. 

Key words: acid base status, rumen pH, urine pH, NABE 

1093 Risk Factors for Bovine Viral Diarrhoea Infection of Dairy 

Farms in New Zealand 

A. Weir, C. Heuer 

Massey University, EpiCentre, Palmerston North, New Zealand 

Objectives of study: New Zealand dairy herds tend to be larger 

(median 250 cows in this study) than in Europe and most are very 

seasonal with all cows in a herd calving over 6 to 12 weeks. Cows are 

not housed and are fed mostly pasture all year round. Calves are often 

grazed away from the main farm from weaning until they return prior 

to calving. There are also a large number of cow movements between 

farms, various forms of cow ownership (share milking system), and 

often rapidly increasing cow numbers. This study evaluated whether 

these and other management factors were associated with BVDV 

infection and immune status. 

Materials and Methods: Initially 268 questionnaires were sent out to 

clients of Eltham District Vet Services with questions relating to 

management factors that may be related to BVD status. From 234 replies, 

206 were selected for bulk tank milk (BTM) testing by PCR for virus and 

by ELISA as a quantitative assessment of antibody levels. A linear 

regression model was built for the S/P ratio of the antibody ELISA. 

Results: There were 21, 51, 50, 70, and 13 herds classed as having high, 

high medium, low medium, low, and nave antibody levels respectively. 

There were 8 PCR positive herds in the high antibody class, and 3 PCR 

positive herds in the high medium class giving a total of 11 PCR positive 

herds. The largest mob size was a better predictor than the herd size and a 

larger mob size was associated with a higher antibody level. The greater 

the proportion of the herd that was bought in the previous season, the 

higher the antibody level. Using fewer bulls for a given number of cows 

was associated with a lower level of antibodies. Using the most common 

BVD vaccine in calves was associated with a lower antibody level in cows 

compared to those that did not vaccinate calves for BVD. Different 

methods of grazing cows over the winter were associated with different 

levels of antibody. Having had BVD diagnosed at any time in the past was 

associated with a higher antibody level. Owner/operators, manager, lower 

order share milker, higher order share milker had the lowest to highest 

antibody levels. 

Conclusions: This study suggests that good agreement exist between 

PCR and antibody tests of bulk tank milk, that infection tends to persist 

over several years, that large mobs maintain infection better than small 

mobs, that risk varies with grazing management and increases with the 

number of cows bought and the number of bulls. 

Key words: BVD, BVDV 

1094 Descriptive Study of Cattle Replacement Practices in an 

Experimental Station for Cattle in the Tropics 

J. Barajas-Rojas 1 , C. Soto Alberti 2 


Microbiology, Mexico, Mexico 

2 USDA-APHIS-VS-CEAH, Microbiology, United States 

The replacement practice of dairy cattle in tropical areas does not 

necessarily follow established criteria or guidelines, and losses may 

differ from those in temperate areas in terms of magnitude and causes. 

There is a need to describe current practices and management 

procedures in livestock production in the tropics. Also, there is a need to 

develop clear and precise replacement procedures through which 

effective control over losses can be established. Costs can be kept down 

by increasing the efficiency of milk production through a planned 

animal health program and by establishing a culling policy in the herd. 

This study describes the management practices of a farm in the tropics 

of Mexico, where records of herd health were kept, but no previous 

replacement program existed.Information was gathered from the 

records of one farm, the Center for Research, Teaching, and Extension 

of Tropical Livestock. Data were collected directly from the field and 

gathered monthly on a 10 year period. Specifically, we were looking at 

replacement due to removal, loss, and cull. Members of each of these 

classifications were then grouped according to infectious, management 

and accident, and productive and reproductive causes. The subjects 

were further classified according to age and genotype. Morbidity and 

mortality data are also presented.The annual removal rate of the female 

population ranged from 2.4% to 36.6% during the 10 years of the study; 

the overall female population removal rate was 6.3%. Removal was due 

45.1% of the time to management and accidents, 37.2% of the time to 

productive and reproductive problems, and 17.7% to infections. In the 

category of management and accident the most important reasons for 

removal were culls to increase the genetic potential of the herd, sudden 

death and bad temper. Low milk production, non-specified problems 

(silent estrus, cyst, reabsortion and repeater breeders), and infertility 

most commonly led to productive and reproductive culling. Mastitis, 

pneumonia and tuberculosis reactors were the most important infectious 

causes for infections. Before the study culling was due largely to 

involuntary reasons, and after to voluntary ones. A better control over 

morbidity now allows for voluntary removal of nonproductive-cows, 

which may increase the genetic potential of the herd. 

1095 Histopathological and Immunohistochemical 

Characterization of Bovine Abortion in Friuli Venezia 

Giulia (Italy) 

E. Pittioni, M. Galeotti 

University of Udine, Faculty of Veterinary Medicine, Department of 

Animal Science, Udine, Italy 

Bovine abortion has an high economical relevance in husbandry, 

therefore the aim of the present investigation is to study this phenomenon 

using histological and immunohistochemical (IHC) methods and to give 

an initial interpretation of it in our region.Pathological samples from 57 

cases of bovine abortion were sent to DIAN (Univ. of Udine) by 

veterinarians between April 2003 and October 2007. Material was 

composed by 47 histological samples (3 samples of placenta; 22 sets of 

foetal organs plus placenta; 22 sets of foetal organs without placenta) and 

12 foetuses (8 without placenta and 4 with placenta). All the samples and 

foetuses were collected in dairy herds in Friuli Venezia Giulia. After a 

preliminary histological examination, a staining method (Gram, Grocott, 

Pizzolato or IHC) was adopted on the basis of pathogens or lesions 

found. Diagnosis was possible on 27 cases (47.4%), no diagnosis was 

done on 30 cases (52.6%). Infectious agents were observed on 26 

(45,6%) samples, being bacterial agents in 15 (26.3%) cases, bovine viral 

diarrhoea virus (BVDV) in 5 (8.8%), bovine herpes virus 1 (BHV1) in 4 

(7%) (2 of which were with mixed infection) and micosis in 2 (7.2%); 1 

(1.7%) abortion was due to renal oxalosis. Bacteria were cause of 

placentitis in 11 (19,3%) cases (4 of which associated with bacterial 

pneumonia and in 1 with septicaemia), in 4 (7%) cases only bacterial 

pneumonia was observed. Antigens related to BVDV were detected by 

IHC in the epithelium of intestinal glands and in some inflammatory 

cells. BHV antigens were detected especially in placenta samples, in 

some cases also in liver and kidney. Fungi were observed exclusively on 

placenta samples. Among 30 undiagnosed cases, 17 (29,8%) had lesions 

but no evidence of any possible agent, 13 (22,8%) had no lesions at all. 

No histological (non suppurative encephalitis and/or cardiomyositis) or 

immunohistochemical evidence of Neosporosis was seen. The most 

damaged organ was placenta, followed by haemolymphopoietic organs 

and intestine. The importance of placenta sampling in case of abortion is 

easily valuable considering that diagnosis was possible on 22 (71%) 

cases of the 31 cases where placenta was available. The relevant 

diagnostic success, compared with diagnostic rate reported in other 

studies, underlines the importance of microscopical investigation for 

diagnosis of bovine abortion. 

Key words: bovine foetus, abortion, immunohistochemistry, 

histopathology 


1096 Determination of Risk Indicators of Postpartum Uterine 

Disease in Dairy Cows - Field study I 

R. Martin, C. Stock, C. Sauter-Louis, R. Mansfeld 

University of Munich, Clinic for Ruminants, Oberschleissheim, 

Germany 

The objective of this study was to identify test parameters and cutpoints 

in the peripartal period allowing the identification of cows at increased 

risk of postpartum uterine disease (PUD) according to the definition of 

Sheldon et al. (2006) (puerperal metritis, clinical metritis, clinical 

endometritis). In eight herds blood serum of 96 cows were collected 

weekly during the period of 2 weeks before expected calving (-2 wk) 

until 2 wk postpartum. Postpartum uterine disease was detected by 

clinical examination up to fifth week postpartum. There were 40 cases 

of PUD (group 1) and 56 cases of no postpartum uterine disease 

(NPUD; group 2). Significant differences were detected in Bilirubin, 

AST, BHBA and Albumin. Predictor cutpoints of these parameters 

were calculated by ROC Curves. The cutpoints of different parameters 

were compared with each other considering sensitivity (sen) and 

specificity (spec) and odds ratio (OR). In PUD higher concentrations of 

Bilirubin from wk -1 to wk 2 (P

counted the number of recent wounds made by bites of bats. Just before 

the twilight a layer of about 5mm of the gel was poured on the whole back, 

from the beginning of the neck to the base of the tail, of all cattle. New 

counting of vampire bats bites were carried out on days 7, 15, 30 and 45 

after the single treatment. Highest counting of cattle recently bitten by bats 

were recorded on days 0 (43/162; 26.5 %) and 45 (31/162; 19.1%), 

followed by days 7 (18/162; 11.1%) and 30 (17/162; 10.4%) (P < 0.04). 

The lowest counting (P < 0.02) was observed on day 15 (7/162; 4.3 %). 

The treatment also reduced the total number of bites/herd (83 d0; 24 d7; 8 

d15; 21 d30; 49 d45) and the average number of bites/bitten cattle (P < 

0.02) from day 7 (1.33 ± 0.5) to day 45 (1.45 ± 0.4) as compared to day 0 

(1.93 ± 1.3). 

Key words: vampire bats, treatment, cattle, warfarin 

1100 Comparison of Some Blood Parameters in Varied Breeds of 

Beef Cows and Their Calves on an Open Farm 

R. Mordak, P. Nowakowski, J. Twardo_ 

Wroclaw University of Environmental and Life Sciences, Animal 

Medicine, Porto, Poland 

Diagnostic monitoring in herd of cattle is very important in maintaining an 

acceptable level of health and productivity in the cows and their calves. It 

is one of the important elements of farm assurance and is necessary to 

demonstrate health and welfare in livestock. In many European countries 

Veterinary Herd Controlling System VHC System is very popular in 

herds of dairy cows (Dairy Herd Controlling System DHC-System) and 

beef cows (Beef Herd Controlling System BHC System) as well other 

breeding species - breeds. It is based on clinical and laboratory 

examinations with blood tests. Knowledge of inner environment and 

homeostasis of animal organisms is crucial for farmers and veterinarians. 

The management of beef cow herds maintained on an open farms is 

particulary difficult. The aim of the study was to compare the values of 

selected blood parameters in three groups (three beef breeds) of clinically 

healthy cows from 50 -70 days post partum and in three groups of their 

calves living together on the same open farm all the year. Trial included 

fifty animals divided in special groups. Hereford groups were consisted 

of 10 cows: group I and their 10 calves; group Ia, Simmental were 

consisted of 8 cows: group II and their 8 calves; group IIa , 

Limousine were consisted of 7 cows; group III and their 7 calves; group 

IIIa. Study included various selected biochemical blood parameters (total 

protein, albumin, creatinine, cholesterol, GOT, GPT, calcium (Ca), 

magnesium (Mg), inorganic phosphorus (P), sodium (Na), potassium (K) 

and chloride (Cl). Significant differences were noted in total protein, 

albumin, chloride, sodium, GOT and calcium between tested groups of 

cows and calves. Detailed data are presented in the paper. 

Key words: beef cattle, blood tests 

1101 Seroprevalence and Risk Factors of Neosporosis in Dairy 

Cows in Western Thailand 

P. Arunvipas 1 , T. Inpunkaew 2 , S. Jittapalapong 2 

1 Faculty of Veterinary Medicine, Kasetsart University, Department 

of Large Animal and Wildlife Clinical Scicence, Nakhon Pathom, 

Thailand 

2 Faculty of Veterinary Medicine, Kasetsart University, Department 

of Parasitology, Bangkok, Thailand 

Livestock development particularly in dairy cows has been hampered 

by low production including milk and slow growth rate due to many 

pathogens including Neospora caninum. N. caninum is the causative 

agent of neosporosis recognized as a major cause of bovine abortion 

around the world and known to have a detrimental effect on bovine 

pregnancy and on milk production. The objective of this study was to 

investigate the seroprevalence and risk factors of N. caninum infections 

in dairy cows of Western part of Thailand. A total of 300 dairy cows 

from 60 dairy herds in three provinces such as Nakhon Pathom, 

Kanchanaburi, and Ratchaburi were randomly selected and examined. 

Sera from 300 cows were collected for antibodies against neosporosis. 

Competitive enzyme-linked immunosorbent assay (c-ELISA, VMRD, 

USA)) was performed as the diagnosis of N. caninum infection. A 

questionnaire of herd management and pet in farm was also obtained in 

each herd. The seroprevalence of N. caninum was 9.7 % (29/300) and 

herd prevalence was 35% (21/60). Ratchaburi has the most positive 

cows with 13% (13/100) and also herds with 45 % (9/20). All positive 

results were seen in all lactations, including pregnant heifer. The results 

of N. caninum infections indicated that dairy cows in Western 

provinces were exposed to N. caninum. There were no significant 

differences in seroprevalence among the parity. The present of dog in 

farms was not significant associated with N. caninum in this study. The 

seen of mouse in farms (OR = 4.1, P=0.05) and other animal present in 

farms (OR 2.9, P=0.007) were significant in this study. Seropositive 

cows trend to have the higher risk of abortion than seronegative 

animals (OR= 2.4, P=0.08) suggesting that N. caninum might be the 

major cause of abortion in dairy cows in Thailand. 

Key words: dairy, neosporosis, risk factors 

1102 Prevalence and Risk Factors for Listeria Species on Dairy 

Farms in Spain 

Mj. Vilar, E. Yus, Fj. Diéguez, Jl. Rodriguez-otero, Ml. Sanjuán 

Instituto de Investigación y Análisis Alimentario, Facultad de 

Veterinaria, Lugo, Spain 

Objectives of study: The aims of the present study were to determine 

the prevalences of Listeria species in on-farm bulk tanks, to identify 

management practices associated with the presence of Listeria species 

in bulk tanks. 

Materials and methods: In 2005 a cross-sectional study carried out in 

98 dairy farms randomly selected in Galicia. Each farmer was 

personally interviewed in farm using a questionnaire about the 

following farm characteristics and aspects of management: silage 

storage, general stabling conditions, milking system and procedure. On 

each farm we collected the following samples: bulk-tank milk, pooled 

faeces from 3 randomly selected, apparently healthy, lactating cows 

and silage. All samples (98 milk samples, 97 faecal samples, 83 silage 

samples) were transported to the laboratory under refrigeration, and all 

analyses were performed within 24 of sampling. Listeria spp. were 

isolated and identificated by standardize method. Silage pH was 

determined at 3 locations in the silage container (upper, middle, lower), 

using a portable pH meter with penetration electrode (Hanna 

Instruments) with an accuracy of < 0.02 pH units. 

Results: Listeria spp. were detected in 16.3% of bulk-tank milk samples. 

Listeria innocua was the species with highest prevalence, detected in milk 

from 7.1% of farms. The next most prevalent species was L. 

monocytogenes, found in 6.1% of tanks. Listeria spp. were detected in 

32.2% of the 180 samples of silage and faeces. In 7.2% of silage samples 

and 3.1% of faecal samples more than one species of Listeria was 

detected. The pH of silage samples from which Listeria spp. were isolated 

ranged from 4.47 to 6.97. We found that ventilation and cleanliness were 

not related to the presence of Listeria in milk. Inadequate management 

during the milking routine favours contamination of milk by Listeria spp. 

We observed a significant relationship between milking system and 

presence of Listeria spp. in milk (P = 0.01, OR = 2.87). 

Conclusions: Correct practices with respect to silage production and 

milking are essential for preventing introduction of Listeria into the 

herd, its spread within the herd, and its entry into milk. The risk of 

contamination of milk by Listeria spp. increased when animals were 

fed low-quality silage, notably silage with pH > 4.5. The prevalence of 

Listeria species and L. monocytogenes in bulk-tank milk was similar to 

that reported in previous studies. 

Key words: dairy cattle, listeria species, epidemiology 

1103 Agreement between Four Commercial Serum ELISAs for 

Detection of Mycobacterium Avium subsp. Paratuberculosis 

Infection in Dairies" 

FJ. Diéguez 1 , S. Gonzalez 2 , A. Menéndez 2 , Mj. Vilar 1 , I. Arnaiz 2 , 

Ml. Sanjuán 1 , E. Yus 1 

1 



2 

Laboratorio de Sanidad y Producción Animal, Xunta de Galicia, 

Lugo, Spain 

Objectives of study: The aim of the present study was assess the utility 

of the different ELISAs currently available for diagnosis of bovine 

paratuberculosis. Specifically, we compared the results obtained in four 

herds with a history of clinical paratuberculosis, using four 

commercially available serum ELISAs. 

Materials and methods: During the first three months of 2006, blood 

samples were obtained from 326 cows from four dairy farms with a 

history of clinical paratuberculosis. These animals were all cows aged 

over one year old on these farms. The ELISAs used are here 

denominated A, B, C and D: SVANOVIR Para-TB-Ab, Svanova 

Biotech, Sweden (A); HerdChek M. Pt., IDEXX, Switzerland (B); 


ParaCheck, CSL/Biocor, Australia (C); and ELISA Paratuberculosis, 

Institute Pourquier, France (D). ELISA A is a lipoarabinomannan 

(LAM)-based indirect ELISA. The other three are based on detection of 

antibodies to protoplasmic MAP antigens and (unlike ELISA A) 

include sample preabsorption with a suspension of the environmental 

mycobacterium M. phlei. After the ELISA testing, faecal samples were 

obtained from 55 animals from the four farms, -namely the 16 animals 

that had tested positive or borderline positive in two or more of the 

ELISAs, a random sample of 35 animals that had tested positive or 

borderline positive in only one of the ELISAs, and a random sample of 

4 animals that had tested negative in all of the ELISAs- for fecal 

cultutre (16 weeks incubation) and PCR analysis (IS900). 

Results: Kappa concordance coefficients in pairwise comparisons of 

the ELISA outcomes ranged up to 0.22 (linear kappa) and 0.25 

(quadratic kappa). When the borderline positives obtained were 

considered as negatives, kappa values remained low (kappa up to 

0.19). Comparative scatterplots indicating the quantitative ELISA 

values obtained for each animal in the sample show rather numerous 

animals with high values in one test and low values in another. 

Coefficients of correlation between these values in pairwise 

comparisons ranged up to 0.45. 

Conclusions: The results of this study confirm the limitations of both 

the currently available serum ELISAs and faecal diagnostic tests 

performed by us for detection of subclinical MAP infections. Some 

previous studies have perhaps over-estimated the utility of the available 

serum ELISAs, by assessing specificity and sensitivity in animals 

whose infection status could be confirmed. 

Key words: MAP, intection, ELISA, culture, PCR 

1104 Impact of and Risk Factors for Mycobacterium Avium 

subsp. Paratuberculosis Infection in Dairy Cattle Farms 

Fj. Diéguez 1 , I. Arnaiz 2 , Mj. Vilar 1 , Ml. Sanjuán 1 , E. Yus 1 

1 



2 

Laboratorio de Sanidad y Producción Animal, Xunta de Galicia, 

Lugo, Spain 

Objectives of study: The aims of this study were to identify risk 

factors associated with Mycobacterium avium subsp 

paratuberculosis (MAP) infection, and to compare mastitis indices 

and reproductive performance at the herd level in farms with 

different serological status. 

Materials and methods: A cross-sectional study carried out in 2004 in 

a total of 101 dairy farms that were randomly selected in Galicia.Each 

farmer was personally interviewed in farm using a questionnaire that 

included questions related to different risk factors that could be 

associated with MAP infection and categorized data about udder health 

and reproductive performance. Cows older than one year were bled and 

samples were analysed for anti-MAP with a commercial ELISA 

(IDEXX). Herds with seronegative cows, or herds with only one 

seropositive cow but no cows with clinical signs consistent with 

paratuberculosis during the previous year, were considered negative (- 

). Herds with 2 - 4 seropositive cows, or herds with one seropositive 

cow and at least one cow culled during the previous year with clinical 

signs consistent with paratuberculosis, were considered positive (+). 

Herds with > 5 seropositive cows, or herds classed as "positive" and in 

which at least two cows had been culled during the previous year with 

clinical signs consistent with paratuberculosis, were considered highly 

positive (++) Multivariable ordinal logistic regression model was used 

for risk factor analysis. 

Results: Logistic regression seemed to indicate as major risk factors 

the use of colostrum from cows with a previous positive diagnosis and 

housing of replacement calves with adult cattle before age six 

months. As regards disease indicators, our results indicate that the 

worse the farm's MAP infection status (negative, positive or highly 

positive), the higher the proportion of farms with high BTSCC; this 

difference in proportions is statistically significant In the case of 

clinical mastitis incidence, significant differences were only observed 

between the negative farms (-) and the highly positive farms (++). As 

regards reproductive performance only days open seemed to differ 

when comparing the negative (-) farms with those of major level of 

infection (++). 

Conclusions: The results of our analysis emphasize the importance of 

adequate biosecurity, with replacement rearing playing a fundamental 

role to reduce the importance of the disease impact in dairies. 

Key words: dairy cattle, epidemiology, MAP, infection 


1105 Realization of the Integrated Veterinary Medicine in the 

Herd Health and Food Animal Production of the Cattle in 

Slovakia 

J. Bires, E. Pilipcinec, M. Vasil, J. Pliesovsky 



Work analyses utilization of elements of integrated veterinary 

medicine from farm on table under provision of herd health 

management of the cattle in the Slovakia. Production and preventive 

veterinary medicine is still exploiting discipline focused on health 

management and production of herd of cattle. It includes creation and 

realization of health programme on farm that are outgoing from activity 

of veterinary surgeon (common clinical observation of the herd, 

analysis of all the relevant information relating to health, productive 

situation, difficulty in herd, resp. and the way of their solution, 

realization of effective prevention). Health programmes in farms of 

cattle are focused on technology of feeding, husbandry, welfare, 

nutrition, metabolism, control of health state of mammary gland, and 

quality of milk, control of health state of locomotory system, 

reproductive functions of cows, health state of calves, epidemiological 

situation and prevention of transimissible infectious disease, etc. The 

important part of the herd health management is situation on farm in 

food safety and quality of produced animals protein and food and 

enviromental safety. Veterinary surgeon during realization of health 

programme on farm with cattle utilizes the elements of 

multidisciplinary position, which are based on the principles of 

integrated veterinary medicine (knowledge of physiology, pathologic 

anatomy, nutrition, animal hygiene and protection of environment, 

welfare, immunology, metabolism, regulatory mechanism, disease of 

cattle, economical aspect of health, public animal health, food safety, 

veterinary legislation, etc.,). 

Key words: integrated veterinary medicine, herd health 

1106 Study the Efficacy of Different Use of PGF 2a and GnRH 

Injection Protocols in Post Partum Dairy Cows 

M. Mohammadsadegh 1 , S. Lotfollahzadeh 2 , A. Bahonar 3 , T. Pazoky 1 

1 

Islamic Azad University, Garmsar Branch, Large Animal Clinic for 


2 

Islamic Azad University, Garmsar Branch, Large Animal Clinical 

Sciences, Garmsar, Iran 

3 

Facutly of Veterinary Medicine, University of Tehran, Food 

Hygiene, Tehran, Iran 

The importance of PGF 2a and GnRH to improve reproductive efficacy on 

post partum periods have been studied by many authors. Reproductive 

management of lactating dairy cows using synchronization ovulation has 

been reviewed by Pursely et al (1997). Experimental manipulation of 

follicular grown (Thatcher et al, 2000) and new clinical uses of GnRH and 

its analogues in cattle have been studied (Thatcher et al, 1993). In order to 

comparing different protocols of hormonal preparation of post partum 

cows 201 non pregnant Holstein cows about after 40 days of parturition 

were divided as follow: (1) In 61 cows only one dose of PGF 2a was 

injected and cows inseminated artificially at estrous. (2) In 31 cows two 

injections of PGF 2a 14 days apart were administered and cows 

inseminated artificially at estrous. (3) (Pre-Select Synch): In 31 cows two 

dose of PGF 2a were injected 14 days apart and 10 days after estrous 

behavior one dose of GnRH, and 7 days after it third dose of PGF 2a were 

injected. The cows were artificially inseminated at estrous observation. (4) 

(Select Synch): In 28 cows PGF 2a were injected 7 days after GnRH 

treatment and artificially inseminated at estrous observation. Statistical 

analysis (Kruskal-Wallis Test,Mann-Whitney Test, Chi- Square Test , 

Student-t Test)were used to analyze parturition to pregnancy (days open ) 

and conception rate in different groups. Results showed that days open 

and conception rate on first service were significantly improved (p

1 

Swedish University of Agricultural Sciences, Clinical Sciences, 

Ruminant Medicine and Epidemiology, Uppsala, Sweden 

2 

Swedish University of Agricultural Sciences, Animal Environment 

and Health, Skara, Sweden 

3 

Swedish University of Agricultural Sciences, Animal Nutrition and 

Management, Uppsala, Sweden 

4 

National Food Administration, Meat Control Division, Uppsala, 

Sweden 

To investigate associations between blood parameters and periparturient 

health status, cows in 40 Swedish dairy herds were blood sampled and 

clinically examined from 4 weeks ante partum (ap) to 7 weeks 

postpartum (pp, 8-26 cows/herd). The status of the cows was classified as 

healthy (n=667), disturbed general condition (n=36), mastitis (n=19) or 

other disease (n=12). Blood parameters analysed were glucose, insulin, 

cholesterol, haptoglobin, glutamate dehydrogenase (GD), nonesterified 

fatty acids (NEFA), betahydroxybutyrate (BHB), aspartate 

aminotransferase (AST) and fructosamine. Associations between status 

and blood parameters were assessed by linear mixed models with 

predictors parity (1, 2, ≥ 3), breed (Swedish Red, Swedish Holstein, 

others), time period (-4 to -2 w ap, -1 w ap, at calving £ 1 day, 1 w pp, 2 

w pp, 3-4 w pp and 5-7 w pp), status and the interaction between time 

period and status. Herd was included as a random effect. Glucose was 

significantly (p

1111 The Prevalence of Hock Lesions in UK Dairy Cattle and the 

Significance of Risk Factors Associated with the 

Development of the Condition 

S. Potterton 1 , M. Green 1 , J. Harris 2 , K. Millar 2 , H. Whay 3 , J. Huxley 1 

1 

University of Nottingham, School of Veterinary Medicine and 

Science, Nottingham, United Kingdom 

2 

University of Nottingham, School of Biosciences, Nottingham, 


3 

University of Bristol, Clinical Veterinary Science, Bristol, United 

Kingdom 

Hock lesions in dairy cattle are widespread around the world. Previous 

studies have recorded a prevalence of approximately 60% in Denmark 

and between 3 and 21% in Switzerland. Prevalence levels from North 

American studies have varied widely from 0 to 91%, according to lying 

surface. In the UK, recent studies found almost 99% of examined cows 

had at least some callused area over a hock, and median proportions of 

ulcerated and swollen hocks of up to 14% and 58% respectively, have 

been observed. Hock lesions are correlated with injuries of the teat, 

carpal joints and skin, increases in lameness score, high somatic cell 

counts, greater incidences of clinical mastitis, and higher culling and 

annual death rates. Differences in the prevalence and severity of hock 

lesions according to the lying surface and cubicle design have been 

identified. Cow-related factors (e.g. bodyweight and condition score, 

breed, and level of milk production), and management variables (e.g. 

herd size, provision for loafing or grazing, and calving season) have all 

been postulated as possible risk factors for the condition. The aim of 

this study was to determine the prevalence of hock lesions in animals 

on dairy farms in the East Midlands of the UK, and the significance of 

potential risk factors in the development of the lesions. A random 

sample of 50 lactating cows on each of 80 dairy farms were assessed 

during a single day visit to each farm over the winter housing period of 

2007/08. The extent and severity of lesions over all aspects of the hock 

were assessed and recorded using a lesion map. Assessments were 

made of lameness score, body condition score, cleanliness, and rising 

behaviour. Days in milk, milk yield, age and parity data were 

collected. Measurements related to the animals environments were 

taken, including dimensions of loafing areas and cubicles, quality of the 

lying surface, and quantity of bedding material provided. Interviews 

were conducted with the farmers in an attempt to establish management 

characteristics and attitudes that may be driving the development of 

this condition. This paper presents data on the location, severity and 

prevalence of hock lesions in UK dairy cattle and identifies factors and 

practices which are associated with the presence of the condition. 

1112 A Comparison of Bovine Serological Profiles of Leptospira 

from two Tropical Regions in Mexico 

L. Moles 1 , J. Torres 1 , J. Cadena 1 , M. Luna 2 , D. Gavaldon 1 

1 Universidad Autónoma Metroopolitana Unidad Xochimilco, 

Producción Agrícola y Animal, México D.F., Mexico 

2 INIFAP, Cenid Microbiologia, D.F., Mexico 

The goal of this survey is to know the seroprofiles of the beef cattle 

form two different regions in the southeastern tropical area of Mexico. 

Microagglutination tests were performed using 10 international 

reference strains and 3 Mexican isolated strain, > 100 titers were 

considered positive. In the first region, 300 bovines belonging to 19 

estates were serologically tested. 84.6% seropositive bovines were 

Hardjoprajitno strain H-89 (60.3%), Hardjoprajitno (49.3%), Panama 

(40.6%), Wolffi (38%), Grippotyphosa (22.3%), Bratislava (21.3%) 

and Tarassovi (9.3%). In the second tested area 203 animals from 6 

estates were analyzed, the range of seropositive animals in the different 

estates was 36 to 89% and the most frequent serovars were 

Hardjoprajitno strain H-89 (67.4%), Wolffi (49.7%), Harjoprajitno 

(43.8%), Bratislava (6.4%) and Tarassovi (5.9%). There is a great 

similarity in the serological profile of leptospirosis in both areas; even 

though the frequency of the serovars is different, they are circulating in 

the same tropical region. It is likely that wild animals may play an 

important role in the maintenance of this serovars. On the other hand, 

despite frequency’s percentage not being the same this is probably due 

to ecological facts. We conclude that bovine leptospirosis is widely 

distributed in the tropical southeastern area of Mexico and the serovars 

identified in this survey point out which bacterin has to been used this 

area to prevent bovine leptospirosis. 

Key words: leptospira, leptospirosis, Bovine, Hardjo, Mexico 


1113 Serological Identification of Antibodies against Leptospira in 

Calves of a Dairy Cattle Farm in Mexico 

L. Moles 1 , J. Hernandez 1 , D. Gavaldon 2 , J. Cadena 1 , J. Torres 1 , 

M. Gonzalez 1 , M. Luna 3 

1 

Universidad Autónoma Metroopolitana Unidad Xochimilco, 

Producción Agrícola y Animal, D.F., Mexico 

2 

Universidad Autónoma Metroopolitana Unidad Xochimilco, DF, 

Mexico 

3 

INIFAP, Cenid Microbiologia, México DF, Mexico 

The aim of this work was to determine when the vaccine against 

leptospirosis should be administered in calves in the Breeding Center in a 

dairy farm region of Tizayuca, in Hidalgo. Sera of 50 calves from (6 days 

to 3 months), 50 from Phase II (4 to 6 months), 30 from Phase III (7 to 8 

months) and 30 from Phase IV (9 to 12 months), were analyzed. The 

microscopic agglutination test (OIE) was used, considering positive the 

sera with titers of 1:100 or more. From the 160 analyzed sera, 9.3% were 

identified positive. In Phase I, 11 (22%) of 50 were found, identifying 8 

(16%) of 50 reactors to the serovar Grippotyphosa; 5 (10%) of 50 to the 

strain H-89 that corresponds to Hardjoprajitno and 3 (6%) of 50 to Wolffi; 

the age of the positive animals went from 6 to 25 days. From Phase II, 4 

(8%) of 50 were identified, 3 (6%) of 50 from the strain H-89 and 1 (2%) 

of 50 from Wolffi; these animals were three months old. There were not 

positive calves in Phases III and IV. During the first 6 days of life, the 

calves remained with their mothers and sucked colostrums, which 

probably explains the presence of antibodies in animals less than 25 days 

of age. Considering that the positive criterion is of 1:100, the sera with 

fewer dilutions might show reactors to a larger amount of serovars found 

in the area. According to this information, the calves are free from 

maternal antibodies from 7 months on. This data suggests the convenience 

of vaccinating at the end of last phase. In other words prior to be 

inseminated and transferred to the breeding area. 

Acknowledgments: Patricia Meléndez for the laboratory support. In 

memoriam Miguel A. Cisneros-Puebla. 

Key words: leptospira, leptospirosis, calves, Mexico, cattle. 

1114 Serological Profile of Cattle from a Semiarid Region of 

Mexican Northwest 

D. Pedroza 1 , M. Luna 1 , V. Banda 1 , F. Salazar 2 , D. Gavaldon 3 , 

L. Moles 3 

1 INIFAP, Cenid Microbiologia, México DF, Mexico 

2 UAEM, Toluca Edo, Mexico 

3 Universidad Autónoma Metroopolitana Unidad Xochimilco, 

Producción Agrícola y Animal, México DF, Mexico 

The purpose of this study was to determine the serological profile of 

beef cattle from a semiarid region from the northwestern part of Mexico, 

149 cows of 3 to 4 years of age from 7 estates were analyzed. All the 

animals had a reproductive disorder data as abortion, infertility and they 

had no positive test to brucellosis. Microagglutination tests were 

performed with 10 different stains of Leptospira international reference 

strains and 3 Mexican strains were added. Titers of ≥ 1:100 were 

considered positive. The percentage of positive animals in each estate 

varied from 47.3 to 100. From all the bovine sera studied 79.8% were 

positive and the titers range was 1:100 to 1:6400. Serovars’ higher 

frequency were Hardjo strain H-89 32.9%, Hardjo strain Hardjoprajitno 

and Bratislava 32.2%, Wolffi 30.2%, Icterohaemorrhagiae strain Palo 

Alto 19.5%, Hardjobovis 18.1%, Canicola 10.7%, Tarassovi and 

Pyrogenes 5.4%, Grippotyphosa 4%, Icterohaemorrhagiae RGA and 

Portland-vere 2%. Up to 6 different serovars were identified in some 

sera. The most frequent serovars in 4 estates were Hardjo strain H-89, 

Hardjoprajitno and Wolffi. In other estates Icterohaemorrhagiae strain 

Palo Alto and Hardjo were very common and in the last one Bratislava 

and Wolffi appeared to be the most important serovars. It is highly 

recommended to consider the serological profiles in order to select an 

efficient vaccine to prevent leptospirosis in this area. 

Key words: leptospira, leptospirosis, cattle, Mexico, Hardjo 

1115 A Retrospective Seroprevalence Study of Bovine 

Leptospirosis in Mexico Considering Ecological Regions 

M. Luna 1 , L. Moles 2 , D. Gavaldon 2 , C. Nava 3 , F. Salazar 4 

1 INIFAP, Cenid Microbiologia, México DF, Mexico 

2 Universidad Autónoma Metroopolitana Unidad Xochimilco, CENID 

MICROBIOLOGIA, MEXICO, D.F., Mexico

3 Clínica Privada, México DF, Mexico 

4 UAEM, Toluca Edo, MEXICO, Mexico 

The study aims to find out the situation of leptospirosis through a 

bibliographic national analysis considering the ecological regions in 

Mexico. The articles were chosen according to the following criteria: a) 

diagnosis technique: microscopic agglutination, b) positive criterion 

1:100 or more, c) time period: 1991-2003, d) type of publication (such 

as thesis, conference reports, well-known and certified magazines) e) 

location according to the state. Duplicated information was excluded. 

The results of frequency and serovars of leptospirosis were reported in 

each state , according to ecological regions: dry and semi-dry, warm, dry 

and humid tropic. The results include 17 of 31 states. Antibodies against 

19 different serovars and seropositive range from 49.7% to 42,779 

serum were analysed. The dry and semi-dry region had a 37.8% 

frequency (range from 31% to 59%), the most prevalent serovars were 

Hardjo genotype hardjoprajitno strain H-89, Hardjo, Wolffi y 

Tarassovi. Warm weather: the average frequency of leptospirosis was 

39.4% (from 22% to 66%). The most prevalent serovars were 

Icterohaemorrhagiae strain Palo Alto, Canicola serovar Portland-vere, 

Bratislava, Pyrogenes; Pomona; strain H-89; Hardjo; Wolffi y 

Tarassovi. Dry tropic: 45.9% (27% to 72%), the most prevalent serovars 

were Wolffi, Hardjo y Tarassovi. Humid tropic: the average frequency 

was 63.8% (from 31.7% to 84.6%), the most prevalent serovars were 

Hardjo genotype hardjoprajitno strain H-89, Hardjo, Wolffi y 

Tarassovi. We conclude that leptospirosis in cattle is endemic in 

Mexico. This suggests a high serological frequency, and that ecological 

factors can determine the prevalence of the disease. The most frequent 

serovars in bovines are Hardjo hardjoprajitno strain H-89, Hardjo 

hardjoprajitno international reference strain, Wolffi and Tarassovi. 

Key words: leptospira, leptospirosis, Mexico, Hardjo, cattle. 

1116 Sanitary Management Related to the Presence of Antibodies 

against Leptospira, Brucella and Neospora in Cows from a 

Dairy Farm 

M. Botello 1 , A. Yaez 1 , J. Torres 1 , L. Moles 1 , D. Gavaldon 2 

1 


Producción Agrícola y Animal, México DF, Mexico 

2 

Universidad Autónoma Metroopolitana Unidad Xochimilco, México 

DF, Mexico 

This study aims to determine serologically the leptospirosis, brucellosis 

and neosporosis sanitary status in a dairy farm of 300 animals, located 

in the central high plateau of Mexico. The sanitary management 

comprises colostrums intake after birth and maternal milk feed during 

the 6 first days, intensive medical care and the appropriate treatment 

when a disease is diagnosed, as well as diagnostic tests when ever are 

required. It is important to point out that most of the calves are retained 

in the dairy farm to replace the remained cows for milk production 

porpoises so there is no animal introduction. There are different zones 

in the farm as the calf breeding, gestation and parturition and the milk 

production areas, so there is no animal contact between one each other, 

the semen used for artificial insemination has a sanitary certification. 

The 300 bovines were sampled for leptospiral diagnosis with 13 

serovars using the microscopic agglutination test considering 1:100 

dilution or higher as a positive titer, brucellosis with bengal rose test 

and ELISA for neosporosis. All the animals tested were serologically 

negative to the 3 diseases studied and it is probably due to the adequate 

sanitary management and the proper installations in the farm that avoid 

the presence and dissemination of the infectious agents of these 3 

diseases at least. It is convenient to mention that an adequate sanitary 

management is necessary to keep a dairy farm free of some infectious 

agents that are evolved in reproductive disorders. 

Key words: leptospira, brucella, neospora, cattle, Mexico 

1117 Lymphocyte Proliferation in Bovine Leukemia Virus- 

Intected Dairy Cows 

F. Souza 1 , M. Azedo 1 , M. Sakai 2 , K. Kieling 2 , R. Weigel 1 , 

B. Sanches 1 , C. Strincagnolo 1 , M. Blagitz 1 , C. Massoco 3 , 

AM. Della Libera 1 




Department of Pathology, Sao Paulo, Brazil 

3 Cornell University, Clinical Sciences, Ithaca, United States 

The purpose of the present trail was to determine the lymphocyte 

proliferation in dairy cows infected with bovine leukemia virus (BLV) 

with distinct lymphocyte profiles in infected animals known as 

alymphocytotic (AL) and persistent lymphocytosis (PL). Thus, 202 

Holstein cows were sera tested by agar gel immunodiffusion (AGID) 

and enzyme-linked immunosorbent-assay (ELISA), 47 days before and 

at the day of the lymphocyte proliferate assay. Besides this, the 

leukocyte profile was evaluated to discriminate these distinct groups 

(AL and PL). The lymphocyte proliferation was done with no 

stimulation (basal), concavalin-A (Con-A) (100 mg/ml) and 

lipopolysaccharide from Escherichia coli serotype O127:B8 (LPS) (250 

mg/ml) when 2x10 6 /ml lymphocytes were plated per well and analyzed 

by flow cytometry. The results on serologic test showed 51% positive in 

both AGID test and ELISA, but in ELISA test about 38% were classified 

as suspect. From these positive animals in both tests 15.60% presented 

LP. From these animals, 15 cows were selected and divided uniformly 

in 3 groups (negative, AL, LP). The mean (± standard deviation) total 

leukocytes counts were 12.04 (±2.51), 11.78 (±0.62) and 35.30 (±11.14) 

x10 9 /L (p

Objectives: To evaluate two different paratuberculosis ELISA test kits 

for serum and milk in 3 German dairy herds with special emphasis on 

the dependence on the herd and the intensity of faecal shedding of 

Mycobacterium avium subsp. paratuberculosis (Map). 

Material and Methods: During a field study of 3 years faecal samples 

as well as blood and milk samples were collected in 3 dairy herds with 

different basic conditions. Samplings were conducted every 5 to 7 

month for 5 times in every herd. Herds A and B had a history of 

paratuberculosis with different prevalence of faecal shedders (30 % vs. 

20 %). Herd C was unsuspicious for paratuberculosis. Faecal samples 

were investigated culturally. 30 animals of herd B which underwent 

necropsy were additionally tested by organ culture. The faecal culture 

results of infected animals were divided into three groups depending on 

the level of shedding. Blood and milk samples were analyzed with 2 

commercial paratuberculosis ELISA test kits. ELISA results of animals 

of herd A and B with at least one Map positive culture result were used 

for the estimation of the diagnostic sensitivities. Diagnostic 

specificities were assessed using the test results of the unsuspicious 

herd C. The impact of the factors herd, level of shedding and repeated 

sampling on test performance was investigated. 

Results: For both ELISA tests sensitivities for blood and milk were 

less than 20%. For all tests a higher sensitivity was shown in herd A. 

Correlation of the actual faecal culture result and the serological test 

result was established. ELISA sensitivities increased significantly 

when animals were heavily shedding Map. A sensitivity of 36.5 % was 

found for blood with ELISA I and milk with ELISA II. In herd A, for 

ELISA I in blood, a sensitivity of 40.2 % was calculated for heavy 

faecal shedders. Specificities between 99% and 100% were reached for 

ELISA II. The specificity of ELISA I was dependent on the inclusion of 

doubtful test results. Specificities between 78.9 % and 94.3 % were 

established when different definitions for true negatives were applied. 

Sensitivity of antibody detection could be increased by repeated 

sampling, whereas specificity was not modified. 

Conclusions: Our results underline that the sensitivity of serological 

tests for sub-clinical paratuberculosis is very limited. Sensitivity and 

specificity estimations for paratuberculosis ELISA are highly 

dependent on the composition of the analysed animal population. 

Key words: antibody detection, specificity, sensitivity, Map 

1120 Types of Bovine Uroliths in Turkey, U.S.A., and Canada 

A. Onmaz 1 , H. Albasan 2 , C. Osborne 2 , J. Lulich 2 , L. Ulrich 2 , 

L. Koehler 2 , V. Gunes 1 , R. Yilmazer 1 

1 

University of Erciyes, Faculty of Veterinary, Internal Medicine, 


2 

University of Minnesota, College of Veterinary Medicine, Urolith 

Center, Minnesota, United States 

There are few reports of quantitative analysis of bovine uroliths in the 

world's veterinary literature. Furthermore, descriptions of uroliths 

appear to be confined to gross observations and chemical spot tests. 

The aims of the study were; 1) to report the quantitative mineral 

composition of 20 uroliths found in the urinary tracts of 1840 Turkish 

cows examined at the abattoir in Kayseri province; 2) to compare these 

results with quantitative analysis of 174 uroliths submitted to the 

Minnesota Urolith Center by veterinarians residing in the USA and 

Canada; and 3) to request that veterinarians who have access to bovine 

uroliths to send them to Dr. Vehbi Gunes along with demographic 

epidemiologic information. The quantitative analysis of each urolith 

from Turkish cows by polarizing light microscopy and infrared 

spectroscopy were carried out in this study. The analyses of the 20 

uroliths revealed that 11 uroliths were composed of magnesium 

calcium phosphate carbonate (55%), 2 uroliths magnesium calcium 

phosphate (% 10), 2 uroliths silica (10%), 1 urolith magnesium 

ammonium phosphate hexahydrate (struvite) (5%), 1 urolith calcium 

carbonate (5%), 1 urolith calcium phosphate (5%), 2 uroliths were also 

classified as compound (10%). One compound urolith was composed 

of a central portion of calcium carbonate surrounded by a shell of silica. 

1 compound urolith was composed of a center of 90% calcium 

phosphate carbonate and 10% struvite, surrounded by a shell of 85 

magnesium calcium phosphate carbonate and 15% struvite. Samples 

(n=174) collected from cows in the USA and Canada were composed 

of the following minerals. Struvite (n = 62, 35.7%), silica (n=34, 

19.5%), magnesium calcium phosphate (n = 19, 10.9%), calcium 

phosphate (n = 16, 9.1%), compound (n = 15, 8.6%), (mixed = 12, 

6.9%), calcium carbonate (n = 7, 4.1%), calcium oxalate (n = 4, 2.4%), 

salts of uric acid (n = 2, 1.1%), drug metabolite (n = 1, 0.6%), and other 


(n = 2, 1.1%). Types of bovine uroliths are different between Turkey 

and North America. It may be related to the type of feed and mineral 

additives. Further controlled studies including feeding and 

management are needed. 

Key words: Bovine, urolithiasis, quantitative, analysis 

1121 Relationship between Periparturient Serum Concentrations 

of Non-Esterified Fatty Acids (NEFA), Beta-Hydroxybutyrat 

(BHBA), Bilirubin and Urea and the Prevalence of 

Postpartum Bovine Endometritis 

T. Kaufmann, M. Drillich, B. Tenhagen, W. Heuwieser 

Faculty of Veterinary Medicine, Freie Universitaet Berlin, Clinic of 

Reproduction, Berlin, Germany 

In the transition period cows undergo substantial metabolic changes. 

Polymorphonuclear neutrophils (PMN), as main uterine defence 

mechanism, are impaired in activity by negative energy balance. 

Deficiencies of PMN function may be responsible for portpartum 

endometritis. Blood serum samples of a total of 130 Holstein-Friesian 

cows were collected 1-7 d ante partum (-7), 1-7 d post partum (+7) and at 

post partum examination (day 28-34) including vaginoscopy. Vaginal 

discharge was regarded as sign for clinical endometritis. If no vaginal 

discharge was found a cytological sample was obtained from the 

endometrium with the Cytobrush technique. The proportion of PMN 

determined the presence of subclinical endometritis. Serum samples were 

analyzed for beta-hydroxybutyrate (BHBA), nonesterified fatty acids 

(NEFA), urea, and bilirubin. Cows with subclinical and clinical 

endometritis in the 5th week after calving had significantly higher NEFA 

concentrations at -7 than healthy cows. Separate analyses for parity 

classes, i.e. primiparous and multiparous cows, revealed that multiparous 

cows with subclinical and clinical endometritis had significantly higher 

concentration of bilirubin at -7 than healthy cows. Furthermore, 

multiparous cows with subclinical endometritis had significantly higher 

concentrations of BHBA +7 than healthy cows and cows affected with 

clinical endometritis. Multiparous cows with clinical endometritis had 

significantly lower concentrations of urea at post partum examination than 

healthy cows. In conclusion, concentrations of NEFA and bilirubin ante 

partum are eligible predictors for the occurrence of clinical and subclinical 

endometritis in multiparous cows. 

Key words: NEFA, endometritis 

1122 Serological, Virological and Haematological Examination of 

Rotavirus Infection in Calves with Diarrhea and Their Dams 

S. Yavru 1 , O. Bulut 1 , S. Sahindurmaz 2 , M. Kale 3 , O. Yapece 4 , 

O. Avce 4 , MK. Albay 2 , F. Pehlivanoglu 2 

1 University of Selcuk, Faculty of Veterinary Medicine, Virology, 

Konya, Turkey 

2 University of Mehmet Akif Ersoy, Faculty of Vet. Med., Department 

of Internal Medicine, Burdur, Turkey 

3 University of Mehmet Akif Ersoy, Faculty of Vet. Med., Virology, 

Burdur, Turkey 

4 University of Selcuk, Faculty of Veterinary Medicine, Virology, 

Konya, Turkey 

In this study, following clinical examination of 3500 cattle and their 

calves from 25 dairy farms 184 cows 2-6 years age and 190 calves 

(three of them are twins) 1-6 months of age with diarrhea were sampled 

for rotavirus presence by ELISA (investigated for antigen in faecal 

samples and antibody in blood serum samples of calves and only 

antibody presence in blood samples of their dams). Additionally, all 

blood samples were examined by hematological methods. 

One hundred sixty seven cows and 144 calves were found seropositive for 

coronavirus. While the high level of antibodies for coronaviruses were 

detected as 32.33% in dams 6years and older ages, the lower level of 

antibodies were detected as 10.18% in ages between 2-3 and 5-6. In the 

calves, Ab to coronavirus were found at the highest level (26.56%) in the 

female calves 5-6 months age but the lowest level (7.81%) antibody was 

found in the male calves 2-4 months of ages. Coronavirus (Ag) was 

detected in only fecal samples of six calves. When the haematological 

parameters were compared between Ab (+) and Ab (-) dams, only White 

Blood Cell (WBC) values were found statistically significant (p

haematological values, no statistical differences were detected. Similarly, 

the haematological parameters of rotavirus Ab (+) / Ag (+) and Ab (-) / Ag 

(-) calves were compared and no statistical differences were detected. The 

haematological parameters were not different between rotavirus infected 

diarrhetic calves and mother, and non-infected animals. Thus, 

haematological examination is not necessary in suspected animals before 

serological and virological testing. 

Key words: Rotavirus, diarrhea, serology, virology, haematology, 

dairy cattle 

1123 Metaphylasis with Florfenicol Based on Temperature at 

Feedlot Entry against BRD under Commercial Conditions in 

Spain 

J. González-Martín 1 , L. Elvira Partida 2 , N. Pérez Villalobos 2 , 

M. Cervio López 3 , E. Calvo López-Guerrero 3 , S. Astiz Blanco, Dipl. 

ECBH 3 

1 

Veterinary Facultiy UCM, Animal Medicine and Surgery, Madrid, 

Spain 

2 

TRIALVET SL, Veterinary Consultancy and Research, Madrid, 

Spain 

3 

Schering Plough Animal Health, Technical Department, Madrid, 

Spain 

Objectives: To evaluate the effectiveness of selective metaphylaxis 

with florfenicol on the basis of a rectal temperature >=39.7 C the day 

that the feedlot reaches the 10% of respiratory disease (day "0"), and 

to asses the efficacy of this temperature for the early diagnostic of 

SRB. 

Materials and methods: The metaphylactic administration of 

florfenicol (SC, 40 mg/kg BW) was investigated in 355 beef calves 

(average weight 268 kg) in four feedlots in Spain. Metaphylaxis (M) 

was applied on Day "0". The ill animals were excluded and the rest, 

n=287, were allocated to one of the experimental groups, using a 

randomization: 

– Group 1: >39.7 C with M 

– Group 2: >39.7 C without M 

– Group 3: 39.7 C and 

metaphylaxis was applied in 43.4% of the animals. Animals with no 

metaphylaxis (n=162) were analyzed to determine the predictive values 

of 39.7 C on Day 0. The result was a 70.4% of diagnosis efficiency, 

with a 52.6% positive predictive value (PPV), and a 72.7% negative 

predictive value (NPV). There were neither significant differences 

regarding the incidence of respiratory disease nor regarding the relapse 

rate between groups 3 and 4 (21.4% vs 27.3% and 5.6% vs 12.8%, 

respectively), either between groups 1 and 2 (36.6% vs 52.6% and 

13.3% vs 10%, respectively). However, the different BRD incidence 

between groups 1 and 2 was numerically important. On Day 30 no 

significant differences were found neither in weight gain (WG) neither 

in average daily gain (ADG) between the different groups. When 

comparing WG and ADG of healthy animals (n=205; 38.8kg and 

1.2kg/day) and diseased animals (n=82; 33.1kg and 1.0kg/day) the 

difference was significant (p=0.02 and p=0.01, respectively). 

Conclusions: From the animal welfare point of view selective 

metaphylaxis based on rectal temperature presents advantages as well 

as good cost/effectiveness ratio and provides appropriate antibiotic use. 

Moreover, the temperature criterion is an objective and useful 

parameter to predict BRD. 

This study was supported by Schering-Plough Animal Health 

1124 A Simulation Model for the Spread of BVD in Germany 

T. Selhorst, J. Gethmann 

Friedrich-Loeffler-Institut, Epidemiology, Wusterhausen, Germany 

Objective: This study was initiated in order to develop a simulation 

model predicting the spread of Bovine Virus Diarrhoea (BVD) virus 

within and between German cattle farms. The model will be used to 

develop efficient BVD elimination strategies. BVD infections in cattle 

cause immense losses. For different countries, these losses vary 

considerably (9.2 C= /cattle/a - 133 C= /cattle/a). Numerous reasons could 

be given for this variation; one plausible determinant is the difference 

between countries with respect to cattle production and management. 

These differences even occur between regions of a country. 

Material and Methods: An individual based, discrete, stochastic 

simulation program was set up, predicting the dynamics of a cattle and 

the dynamic of a BVD infection. The model consists of different 

modules, two of them being module Cattle and module Farm. Module 

Cattle predicts the fate of individual cattle, module Farm combines 

cattle in a farm and predicts the spread of the disease within farms and 

between farms via trade. Data describing the unique cattle production 

and management conditions in Germany were used to set up the 

modules. These data are stored in the national cattle database 

(Herkunftssicherungs- und Informationssystem für Tiere, HI-Tier), 

consisting of more than 10 million cattle CV’s, and the information 

about trade between farms from the year 2001 onwards. Using this 

database, the most veritable data for the simulation of the cattle herd 

and BVD dynamics in Germany were used. 

Results: The model developed so far has been validated by national 

experts, and will be further checked with the help of the observed BVD 

dynamics in certain herds. If the model will pass these checks, it will be 

used to develop guidelines for the implementation of cost efficient 

BVD elimination programs. 

Conclusions: In order to develop cost efficient strategies for the 

elimination of the BVD virus within German cattle herds, it was 

decided to develop a general simulation module predicting the specific 

cattle herd and trade dynamics observed in Germany. The BVD 

dynamics has been linked with this dynamics. This linkage will assist 

disease managers in developing cost efficient BVD elimination 

strategies. The models excess value is given by its applicability to other 

cattle diseases. For this purpose, the disease specific aspects must be 

considered. 

1125 Prevalence of Fluorosis in Buffaloes in Punjab State of India 

J. Singh 1 , S. Uppal 2 , K. Dua 2 , S. Randhawa 2 , C. Randhawa 2 

1 Veterinary Officer, Animal Husbandry Deapartment, Sangrur, India 

2 Guru Angad Dev Veterinary and Animal Sciences University, 

Veterinary Clinical Services Complex, Ludhiana, India 

In India there are endemic fluorosis areas where clinical signs are 

manifested in animals due to prolonged consumption of contaminated 

ground water (hydrofluorosis) or fluoride rich brick kiln ash 

contaminated fodder (non-hydrofluorotic area). An epidemiological 

survey on fluorosis in buffaloes was conducted in hydrofluorotic and 

non-hydrofluorotic areas of Punjab. A total of 216 buffaloes were 

randomly selected from three types of areas. In group I, 155 buffaloes 

were selected from 8 villages in hydrofluorotic areas without any brick 

kilns in their surroundings; in group II, 28 buffaloes from 3 villages in 

non-hydrofluorotic areas having brick kilns, while in group III, 33 

buffaloes were selected from 2 villages in hydrofluorotic areas having 

brick kilns. Ten healthy buffaloes between 3-12 years of age from 

non- fluorotic belt having water and fodder fluoride levels of 1.06ppm 

and 0.183ppm, respectively were selected as control group. Plasma, 

fodder and ground water fluoride analysis was done using fluoride 

electrode by standard methods. Significantly (p0.3 ppm). The study indicates widespread prevalence of 

fluorosis in Punjab state in endemic areas and in non-endemic areas in 

the vicinity of brick kilns. 

Key words: buffalo, fluorosis 


1126 Relationships between Paratuberculosis Sero-status and 

Milk Production, SCC and Calving Interval in Irish Dairy 

Herds 

K. Hoogendam 1 , E. Richardson 2 , J. Mee 2 

1 

Van Hall Instituut, Agora 1, Leeuwarden, Netherlands 

2 

Teagasc, Moorepark Dairy Production Department, Fermoy, Co. 

Cork, Ireland 

The objective of this study was to investigate the impact of 

paratuberculosis sero-status on milk yield, fat, protein, somatic cell count 

and calving interval in a sample of Irish dairy herds. Serum from all 

animals (n=2,602) over twelve months of age in 34 dairy herds was tested 

for paratuberculosis antibodies using an ELISA (Pourquier). Herds were 

categorised by sero-status into positive, non-negative and negative, where 

a positive herd contained two or more positive cows, a non-negative herd 

contained only one positive cow and a negative herd contained no positive 

cows. The production and reproduction records of the current lactation 

(year of test) were compiled from the Irish Cattle Breeding Federation 

database. Data at animal, parity and herd-level were analyzed by multiple 

regression using general linear models. The true animal-level prevalence 

of paratuberculosis was 2.8%. At the herd-level, 33% of herds had two or 

more positive cows and 50% of herds had at least one positive cow. 

Positive herds (n=129 cows/herd) and non-negative herds (n=81) were 

larger than negative herds (n=72) (p70 positive), the sampling frequency, the low seropositive animallevel 

prevalence and the unavailability of clinical data on Johne’s disease, 

these results are not surprising. This was the first study to examine the 

relationships between paratuberculosis sero-status and production in a 

sample of Irish dairy herds. 

Key words: paratuberculosis, ELISA, milk production, dairy herds 

1127 Clinical Surveys in Acute Lead Poisoning in Dairy Cattle 

S. Catania 1 , O. Parolin 2 , G. Binato 1 , M. Corr 1 , E. Schiavon 1 , 

M. Merenda 1 , D. Bilato 1 , L. Iob 1 

1 

Istituto Zooprofilattico Sperimentale Delle Venezie, Legnaro (PD), 

Italy 

2 

Veterinary Practitioner, Legnaro (PD), Italy 

Objectives of study: Lead intoxication in dairy cattle is widely 

reported (Lemos 2004; Ozmen 2004) and causes mainly neurological 

malfunctions and loss of production (Frape 1984; Radostits 2006). 

Diagnosis is based on clinical signs and level of lead in tissues. In a 

dairy cattle farm, milking cows and calves were accidentally fed with a 

truck battery. 

Materials and methods: After a sudden loss in milk production, 

anamnesis detected the accidental inclusion in the food of an exhausted 

truck battery, ingested by 52 milking cows and 4 calves. Lead 

poisoning was suspected. Clinical alterations were recorded and blood 

samples were taken to determine lead level, using atomic absorption 

spectrometry-GFAAS. Symptoms were monitored on days 7, 12 and 

15. A therapy based on Ca-EDTA was given to the 6 animals 

presenting symptoms (Radostits 2006) 

Results: Five cows and one calf showed clear clinical signs on day 7: 

no feeding, no ruminating, intense salivation, muscular tremors, teeth 

baring with clicks, mydriasis, fear, hyperesthesia and rise of heart and 

respiratory rate. Two animals had blackish fetid diarrhea anticipated by 

stipsis. The loss of milk production was about 85%. A cow and a calf 

were blind. The remaining 47 cows showed mild excitement and fear. 

Blood sampling was difficult and lead levels ranged between 0.39 and 

0.76 mg/l.On day 10, 7 more cows had milder symptoms: decreased 

food assumption, variable loss of milk production, irregular 

rumination, nervousness, fear, mydriasis and tremors.One of the 

symptomatic cows died on day 12, in spite of therapy. The autopsy 

revealed haemorragic enteritis, metallic fragments in the reticulum and 

a greyish coat in omasum. During the same day, 3 more calves showed 

weawing and alterations while standing.On day 15 the group gradually 

recovered rumination, visual functionality, milk production. One more 

death was reported, due to ab-ingestis pneumonia after therapy. 


Conclusions: Clinical data and lead levels confirm acute lead 

intoxication. Symptoms arose in two moments, due to the presence of 

two forms of lead inside the battery: metallic lead and lead sulphate. In 

particular, lead sulphate proves to be more bioavailable with 

consequent acute symptoms. Metallic lead is less absorbable and 

symptoms can appear later. Metallic fragments do not disperse 

uniformly in the food, and tend to sediment. This could explain the 

arise of symptoms in calves, that were fed with leftovers of cow 

food. Bibliography is available upon request. 

Key words: lead poisoning, GFAAS, syntomps, diagnosis 

1128 Bacteriological and Serological Study on the Infection of 

Mannheimia (Pasteurella) haemolytica in Cattle in Ahvaz 

(Southwestern of Iran) 

MR. Haji Hajikolaei 1 , A. Rasoli 1 , M. Ghorbanpoor 2 , 

MR. Saifiabad-shapouri 2 , D. Ebrahimkhani 3 

1 



2 


Department of Pathobiology, Ahvaz, Iran 

3 

Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, 

Iran 

Pneumonic pasteurolosis of cattle (shipping fever pneumonia) which 

caused by Mannheimia (Pasteurella) haemolytica and Pasteurella 

multocide is a major cause of economic loss in the feedlot industry. The 

disease occurs most commonly in young growing cattle from 6 months to 

2 years of age. The frequency of isolation of Pasteurella spp. from the 

nasal passage of normal healthy unstressed calves is low but increases as 

the animals are moved to an auction mart and then to a feedlot. In order to 

investigate the prevalence of Mannheimia (Pasteurella) haemolytica 

infection in cattle in Ahvaz (Southwestern of Iran) bacteriological and 

serological study was carried out on 250 slaughtered cattle at Ahvaz 

abattoir. Nasal and nasopharyngeal swabs and blood samples were taken 

from each cattle after slaughter. Nasal and Nasopharyngical swabs were 

cultured in blood agar and incubated at 37 °C for 24-48 hours. The 

suspected bacterial cultures were processed for isolation of multocide and 

haemolytica following routine bacteriological techniques. Sera were 

tested by indirect hemagglutination test (IHA) to reveal antibodies against 

this organism. M. haemolytica was isolated in 1.6% cattle. Statistical 

analysis showed that there were no any relation between age and sex with 

bacterial infection. Serological studies showed that 71.6% tested sera 

contained antibody (titer ≥ 1/16) against M. haemolytica and no relation 

between age or sex with serological results. 

Key words: Mannheimia haemolytica, cattle, Ahvaz, Iran 

1129 Tick-borne Diseases in Dairy Cattle: Conditioning Factors in 

a Mediterranean Endemic Area 

L. Ceci, P. Paradies, D. De Caprariis, F. Iarussi, M. Sasanelli, 

G. Carelli 

University of Bari, Animal Health and Welfare, Valenzano (Bari), 

Italy 

Objectives of study: The aim of the study is to asses the incidence of 

Tick Borne Diseases (TBDs) in a dairy herd located in an endemic area 

(southern Italy). Furthermore the possible role of conditioning factors 

in determining the onset of clinical forms of disease is discussed. 

Materials and Methods: A dairy herd was monitored for a period of 

32 months. The farm, located in Apulia (southern Italy) is composed of 

140 Holsten Friesan and Brown breed cattle of different ages. Several 

clinical cases of anaplasmosis by Anaplasma marginale and babesiosis 

by Babesia bigemina have been registered over the years in the same 

herd along with the presence of Theileria buffeli. In the herd oxytocin 

was usually administered for the entire period of lactation to facilitate 

milk ejection and increase production. All animals of the herd were 

monitored through clinical examination and bleed for haematological 

exams and molecular investigations. 

Results: 38 clinical cases of TBDs were observed during the study; in 

particular an outbreak of babesiosis from B. bigemina (24 cases) was 

observed in December, corrisponding to a sudden decrease in 

temperature. In the following 24 months 14 clinical cases of 

anaplasmosis were observed in cattle during post-partum in different 

seasons. In the animals showing simptoms of anaplasmosis A. 

marginale was revealed at microscopy in 10 animals, A. marginale and 

T. buffeli in 3, A. marginale, B. bigemina and T. buffeli in 1.

Conclusion: The high incidence of TBDs here reported suggests the 

importance of some factors in the development of clinical forms in 

endemic areas. We suppose that the rapid temperature decrease have 

played a determining role in the onset of the outbreak of babesiosis as 

well as the oxytocin administration and/or the stress of post-partum in 

the development of cases of anaplasmosis. Dairy cattle living in 

endemic areas for the TBDs can develop clinical simptoms starting 

from a latent infection. The delicate balance that is established between 

parasite and host is influenced by several intrinsic and extrinsic factors. 

These factors could play a decisive role in the onset of clinical forms of 

the disease in endemic areas. Thus, their occurrence is not only due to 

the presence of the pathogen and vectors but, significantly, to 

concomitant conditioning factors. 

Key words: dairy cattle, tick-borne diseases, Anaplasma marginale, 

Babesia bigemina, conditionig factors 

1130 Serological Determination of Leptospirosis and Brucellosis 

in Bovine Fetuses 

MP. Estrada 1 , J. Torres 1 , LP. Moles 1 , P. Melendez 1 , 

L. Benavides 1 , R. Soto 2 , L. Hernandez 3 

1 Universidad Autonoma Metropolitana-Xochimilco, Producción 

Agrícola y Animal, MEXICO, D.F., Mexico 

2 Cuenca Lechera de Tizayuca Hidalgo, México, México DF, Mexico 

3 INIFAP, Cenid Microbiologia, México D.F., Mexico 

Objective: The aim of this work was the serological determination of 

leptospirosis and brucellosis in bovine fetuses from the dairy complex 

at Tizayuca (DCT) in the Hidalgo State of Mexico, with an 

approximately 29.000 cows in production. 

Materials and Methods: We analyzed samples of sera from 17 

aborted fetuses and 15 fetuses still in utero of dead cows, from different 

farms in DCT. Also 133 non pregnant cows were sampled from the 

same farms where the fetuses’ samples were obtained. The 

antileptospiral titration of the sera was performed by the micro 

agglutination (MAT) technique using 13 serovars of Leptospira as 

antigens. The titer was considered as the maximal dilution of sera in 

which 50% of agglutination of the antigens was observed. For the 

definition of anti brucella antibodies the radial immunodiffusion (RI) 

test was used with an antigen preparation of Brucella mellitensis. 

Results: From the 32 analyzed fetuses only one reacted against two 

serovars of Leptospira, at the dilution 1:20 with Hardjo Hardjoprajitno 

strain H-89 isolated in DCT in 1989 and at 1:40 dilution against Wolffi, 

the fetus was aborted at the 240 th day of gestation. Seventy four percent 

of the sera from the producing cows reacted against one or more 

serovars of Leptospira, the most frequent ones were: Hardjoprajitno H- 

89 (23.6%), Hardjo (17.5%) and Icterohaemorrhagiae (12.2%). Only 

the serum of one in utero fetus of approximately 180 days of gestation 

reacted against the Brucella antigen. 

Conclusions: It is known that bovines are immunocompetent since 160 

days of gestation. The OIE have suggested that positive titers of 

specific antibodies against Leptospira or Brucella in the sera of bovine 

fetuses may be taken as a definitive diagnosis of these infections. 

Therefore our findings indicate that the two seropositive fetuses were 

produced by an active infection of Brucella and Leptospira. 

Key words: determination, leptospirosis, brucellosis, bovine fetuses 

1131 Evaluation of Vaccine Preparations for Bovine Leptospirosis 

by the Micro Agglutination and Growth Inhibition Tests 

C. Cedillo 1 , K. Morales 1 , H. Castillo 1 , JI. Torres 1 , L. Benavides 2 


Agrícola y Animal, Mexico, Mayotte 

2 Universidad Autonoma Metropolitana-Xochimilco, Sistemas 

Biologicos, MEXICO, D.F., Mexico 

We compared the microscopic agglutination test (MAT) and the growth 

inhibition test (GIT) in the evaluation of the antibody production elicited 

by three vaccine preparations from different serovars of the Sëjroe 

serogoup of Leptospira. Forty Holstein Friesian calves seronegatives 

against Leptospira were divided in four groups of ten individuals each. 

Group A was inoculated with a trivalent bacterin prepared with 

Hardjoprajitno strain H-89 isolated in Mexico, Hardjo and Wolffi; Group 

B with an univalent bacterin from Hardjoprajitno strain H-89; Group C 

with a commercial bacterin from Hardjo Bovis and Group D was the 

control group, unimmunized. The animals were immunized 

subcutaneously on days 0 and 15 and were bled on days 0, 7, 14, 21 post 

inoculation (pi). For the evaluation of the capacity to elicit an immune 

response we used the MAT with the homologue serovars as antigens. In 

the GIT, 3 ml of modified Cox medium with 0.3 ml of sterile rabbit serum, 

0.03 ml of the sterile problem serum and 0.3 ml of a culture of the strain 

Hardjoprajitno H-89 of Leptospira, or 0.3 ml strains: Hardjoprajitno H- 

89, Hardjo and Wolffi. No problem serum was added to the control tubes. 

Cultures were incubated a 28°C and observed at the dark field microscope 

periodically. The A and B groups stimulated antibody production from 

day 7 pi, the antibodies decayed at day 21. Fewer animals inoculated with 

the commercial bacterin responded and at lower titers (two to three 

dilutions less), compared with those of groups A and B. In group D, the 

control animals were all negatives. With both tests MAT and GIT the 

effect of vaccination was statistically significant (p < 0.0001). The IEO 

recommends the utilization of virulent strains to test the efficiency of the 

vaccines in development but this type of tests are risky and the 

maintenance of such strains is difficult and costly; our results reinforce the 

value of GIT and MAT as tests for evaluation of the inmunogenicity of 

vaccine preparations. 

Key words: evaluation, vaccine, leptospirosis, MAT, GIT 

1132 Genotyping of Mycobacterium Avium Subsp. 

Paratuberculosis Isolates Based on Mycobacterial 

Interspersed Repetitive Units (MIRU) Methods 

A. El-Sayed 1,2 , S. Natur 2 , W. Wolter 2 , M. Zschöck 2 , A. Hassan 2 

1 Faculty of Veterinary Medicine, Cairo university, Institute of 

Internal Medicine and Animal Infectious Disease, Cairo, Egypt 

2 Landesbetrieb Hessiches Landeslabor, Institute of Internal Medicine 

and Animal Infectious Disease, Giessen, Germany 

Mycobacterium avium subsp. paratuberculosis (MAP) is the 

etiological agent of severe gastroenteritis in both ruminants (Johne's 

disease) and human (Crohn`s disease). The molecular diversity of 109 

MAP isolates were investigated in the present study. Six different 

Mycobacterial interspersed repetitive units (MIRU) methods were 

applied to classify the MAP isolates in to subgroups. The genotypic 

characterization revealed the presence of 25 different profiles and two 

of them dominate among investigated strains. The first profile could be 

detected in 41 isolates (37,6%) with amplicon size of 300, 200, 350, 

200, 210 and 210 bp for the primer pair (292-F, 292-R); (X3-F, X3-R), 

(25-F, 25-R); (3-F, 3-R); (7-F, 7-R) and (47-F, 47-R), respectively. The 

second profile is present in 28 isolates (25,7%) with amplicon size of 

350, 250, 350, 200, 190 and 210 bp for the same primers, respectively. 

In addition, 16 profiles were represented by one strain per profile. 

Using different MIRU methods in the present study showed promising 

results for improvement of epidemiological studies of MAP. 

Key words: Mycobacterium avium subsp. paratuberculosis, 

Mycobacterial interspersed repetitive units (MIRU), Johne's disease, 

Crohn`s disease, epidemiology 

1133 Detection of Mycobacterium Avium Subsp. Paratuberculosis 

in Manure, Environmental Samples and Milk Filters of 

Dairy Herds 

A. Hassan, S. Natur, A. El-Sayed, W. Wolter, M. Zschöck 

Landesbetrieb Hessiches Landeslabor, Abteilung Veterinärmedizin, 

Giessen, Germany 

Mycobacterium avium subsp. paratuberculosis (MAP) is a robust 

worldwide spread microorganism which causes specific incurable chronic 

enteritis in cattle and many animal species. The objective of this study was 

the detection of MAP in manure, in environmental and milk filter samples 

of dairy herds in Hesse region, Germany. Manure samples (n = 60), 

environmental samples (n = 19) and milk filter samples (n = 60) were 

collected from 60 herds which are known serologically to be MAP 

positive and MAP negative. All samples were cultured using Herrold's 

Egg Yolk Medium (HEYM) after a decontamination step with 

Hexacetylpyridium chloride (HPC 0.75%) for 18 h. Among all herds 

investigated five manure (8.3%), two environmental samples (10.5%) and 

two milk filter samples (3.3%) showed typical growth of MAP. The 

molecular identification of the isolates could successfully be performed 

using species specific PCR methods based on insertion sequence IS900 

and the gene f57. According to the present results manure, environmental 

and also milk filter samples seem to be of importance for management 

strategies to control paratuberculosis in dairy herds. 

Key words: Mycobacterium avium subsp. paratuberculosis, 

environmental samples, disease control 


1134 A Comparison between Two Calf Breeding Systems and its 

Influence in Weight Gaining at Weaning 

ML. Botello, JI. Torres, LP. Moles, D. Galvadon 

Universidad Autonoma Metropolitana-Xochimilco, Producción 

Agrícola y Animal, México D.F., Mexico 

The goal of the research is to contrast two breeding systems as a well as 

the nutritional value of two nutriments given to calves in order to 

determine the weight gaining at weaning. The study was carried out in 

two Holstein dairy farms with similar environments and managerial 

strategies. The colostrum given during first three days colostrum from 

one, two and three parturitions cows was given to the group (A) during 

the first three days after birth. The calves were provided with 

commercial food from this point until the 80 th day. Meanwhile group B 

was fed colostrum from three parturition cows for two days and 

commercial food up to the 90 th days. A bromatological analysis was 

carried out to colostrum, whole milk, commercial food and dried 

alfalfa. In order to determine the average of body weight gained 15 

newborn and 15 weaned calves were measured. The colostrum’s 

infrared spectroscopical analysis gave the following results: group A 

had 12.03% proteins, 3.46% fat, 3.45% lactose and total solids 19.63%. 

Whereas group B displayed 19.86% proteins, 3% fat, 3.22% lactose 

and 27.48% total solids. The proximal analysis of the commercial food 

and dried alfalfa showed for the group A 18.35 % crude protein (CP), 

19.05% crude fat (CF), 0.13% crude fiber (CFi) and pellet milk’s 

substitutes 28% CP, 10% CF and 1% CF, commercial food 18.97% CP, 

0.10.86% CF, 3.64% CFi, 71.66% nitrogen free elements (NFE) and 

5.69% ashes (A), dried alfalfa 24.5% CP, 0.0892% CF, 20.79% CFi, 

43.06% NFE, 11.54% A. The group B milk substitute 20% CP, 21% 

CF, 0.15% CFi, dried alfalfa 25.6% CP, 2.7% CF, 20.1% CFi, 36.1% 

NFE, 11.9% A. The commercial food reports 25% CP, 3% CF, 3% CFi. 

The weight gained in group A was 51.9 Kg and group B 62.250 Kg. 

Group B complies better with the minimal nutritional requirements 

than group A. The data suggest that a good quality milk substitute 

works better than whole milk because satisfies the minimal 

requirements at a lesser cost. The former secures quick ruminal 

development al weaning. Getting to the adequate body weight requires 

good management during lactation stage. It is always desirable to 

supply high quality colostrum during the calf’s growing stage. 

Key words: breeding systems, weight gaining, calves 

1135 Anti Leptospiral Antibodies in Equines and Canines in an 

Endemic Area of Bovine Leptospirosis 

C. Mendez 1 , JI. Torres 1 , P. Melendez 1 , L. Benavides 2 


Agrícola y Animal, México DF, Mexico 


Biologicos, México D.F., Mexico 

We pretended to identify specific antileptospiral antibodies in equine 

and canine sera from an area of endemic bovine leptospirosis caused 

mainly by serovars Hardjo and Tarassovi. We essayed 222 sera from 

bovines: 70 in gestation, 83 non gestating and 69 calves; 24 equine sera 

and 6 canine sera, all from Soto la Marina, Tamaulipas state in Mexico. 

The antileptospiral titration of the sera was performed by the micro 

agglutination (MAT) technique using thirteen serovars of Leptospira as 

antigens: Double dilutions of the sera starting in 1:50 were done, the 

titer was considered as the maximal dilution of sera in which 50% of 

agglutination of the antigens was observed. The results with the bovine 

sera were: 48 (68.57%) positives from the pregnant cows, to one or 

more serovars, the more frequent ones: Hardjo strain H89, 40 

(57.14%); Hardjo prajitno 36 (51.42%), Wolffi 34 (48.57%) and 

Tarassovi 18 (25.71%). From the non gestating ones 52 (62.65%) were 

positives to one or more serovars, principally to Hardjo H89 38 

(45.78%) and Wolffi 18 (21.68%). The calves had 79.71% (55) of 

seropositivity, being Hardjo prajitno y Hardjo H89 the highest (51 or 

73.91%) and Wolffi 28 (40.58%). The horse sera reacted against one or 

more serovars, 22 positives (91.6%) mainly against Tarassovi 

11(54.83%), Hardjo prajitno 9 (37.5%), Bratislava 7 (29.16%) and 

Canicola 7 (29.16%). The entire dog sera were positives to one or more 

serovars especially to Canicola 6 (100%), Portland-Vere 6 (100%), 

Icterohaemorrhagiae Palo Alto 6 (100%), Icterohaemorrhagiae 4 

(66.6%) and Pyrogenes con 3 (50%). As in other parts of the Country, 

we confirmed the incidence of bovine leptospirosis in Soto la Marina 

specially the serovar Tarassovi and the related ones from the Serjöe 


Serogroup. Since the equine sera reacted mainly with serovar 

Tarassovi, we need to establish the importance of this species as a 

source of infection and transmission of this strain of Leptospira. This 

type of studies are relevant because of the close contact that horses 

have with bovines in this region and since it is not known the role that 

this species play in the transmission of serovar Hardjo to the bovines. 

Key words: antileptospiral, antibodies, equines, caninen and bovines 

1136 Antileptospiral Antibodies in Bovine and Human 

Populations Ina Dairy Farm in Tizayuca, Hidalgo Mexico 

JI. Torres 1 , L. Benavides 2 , P. Melendez 1 , D. Galvadon 1 , ML. Botello 1 


Agrícola y Animal, Mexico, D.F., Mexico 


Biologicos, México D.F., Mexico 

The objective of this work was to compare the serological profiles 

against leptospira in the human and bovine populations from a dairy 

farm near Mexico City. From an approximate population of 27,000 

milk producer bovines from the dairy farm of Tizayuca (TDF) in the 

Hidalgo state, Mexico; 240 bovine sera and 45 sera from farm workers 

and veterinarians, with one to 20 years of work in the farm, were 

sampled. The antileptospiral titration of the sera was performed by the 

microagglutination (MAT) technique using twelve serovars of 

Leptospira as antigens: Bratislava Jes-Bratislava, Canicola Hond 

Utrecht IV, Portland-vere Sinaloa ACR, Grippotyphosa Moskva V, 

Icterohaemorrhagiae RGA, Icterohaemorrhagiae Palo Alto, Pomona 

Pomona, Pyrogenes Salinem, Hardjo Hardjobovis LT 1085, Hardjo 

Hardjoprajitno H-89, Wolffi 3707 and Tarassovi Perepelitsin. The titer 

was considered as the maximal dilution of sera in which 50% of 

agglutination of the antigens was observed. From 240 bovines sera, 

61%(147) reacted against on or more of the serovars used as antigens in 

the MAT, the most frequent was Hardjo Hardjoprajitno H-89 (isolated 

from TDF in 1989) with 42% (101); Hardjo with 27%(65); Wolffi con 

21%(51) and Tarassovi con 16%(39). In the 45 human sera analyzed 

we could not detect any reaction with the serovars used in the MAT. 

From the reactivity observed in the bovine sera against the serovars 

Hardjo and Wolffi from the serogroup Serjoe of Leptospira indicate 

that these serovars are in active circulation in the bovine population of 

the farm. The Hardjo serovar has been reported as the principal 

responsible of abortions and mortinates, weak calves at the birth, 

infertility and low milk production. In humans the main source of 

infections are the rodents and occasionally other mammals as may be 

the bovines. The fact that in this study the serological profiles of 

antileptospiral antibodies of bovines and men in the TDF did not 

coincide led us to think that L. Hardjo is well adapted to the bovine but 

not to man, even when he is working in close contact with active 

infected animals. 

Key words: antileptospiral antibodies, bovine, human, Mexico 

1137 Association of Bovine Viral Diarrhea Virus (BVDV) and 

Bovine Herpesvirus-1 (BHV-1) in Abnormal Cervical 

Mucous Discharge and Fertility in Cows and Heifers 

S. Yavru 1 , M. Kale 2 , A. Ayhan 3 , O. Bulut 4 , MS. Gulay 5 , 

O. Yapkc 4 , F. Pehlivanoglu 6 

1 

University of Selcuk, Faculty of Veterinary Medicine, Virology, 

Konya, Turkey 

1 

University of Mehmet Akif Ersoy, Faculty of Vet. Med., Virology, 

Burdur, Turkey 

3 

University of Mehmet Akif Ersoy, Faculty of Vet. Med., Department 

of Artificial Insemination, Burdur, Turkey 

4 

University of Selcuk, Faculty of Veterinary Medicine, Virology, 

Konya, Turkey 

5 


of Physiology, Burdur, Turkey 

6 


of Veterinary Microbiology, Burdur, Turkey 

The purpose of the present study was to describe whether Bovine Viral 

Diarrhea Virus (BVDV) or Bovine Herpesvirus-1 (BHV-1) in 

abnormal cervical mucous discharge (A-CMD) has effects on fertility 

of cows and heifers. For this purpose, after visual examination of CMD 

of the animals in estrus (n=222), animals having clear discharges (68 

cows, 38 heifers) with normal viscosity and without bad odor were 

grouped as normal cervical mucous discharge (N-CMD) group. The

others were grouped as A-CMD (84 cows, 32 heifers) group. All 

animals were inseminated with BVDV and BHV-1 free frozen semen. 

Samples were tested by enzyme linked immunoassay (ELISA) for 

antigens (Ag) of BVDV and BHV-1. BVDV infection was not 

associated with A-CMD or conception rates (CR) for cows or heifers, 

and BHV-1 Ag were not detected in any samples. Thus, BVDV did not 

change the characteristics of observable CMD in the current study and 

presence of BVDV must be revealed serologically. 

Key words: cervical mucous discharge, BVDV, BHV-1, fertility 

1138 Cefquinome Disk Susceptibility Test and Interpretative 

Criteria for Cefquinome in vitro Activity Testing against 

Veterinary Pathogens 

M. Rose 1 , J. Swinkels 2 

1 Intervet-Schering Plough, R&D, Schwabenheim, Germany 


Objectives: Currently, cefquinome susceptibility testing disks 

containing 10 g cefquinome are being used in veterinary laboratories all 

over the world. These disks have an inhibition zone of 18mm. In order 

to explore optimization of the antibiotic load in cefquinome disks, a 

study based on the CLSI M37-A2 standard has been conducted to 

evaluate disk diffusion data for 30 g cefquinome disks. 

Materials and Methods: The MICs of cefquinome were generated by 

broth microdilution and inhibition zone diameters were defined by disk 

diffusion against a collection of gram-negative and gram-positive 

isolates of aerobic veterinary pathogens, including fastidious organisms. 

In total 702 isolates, including 120 M. haemolytica, 117 P. multocida, 

52 A. pleuropneumoniae, 90 H. somni, 76 E. coli, 18 K. pneumoniae, 10 

S. marcescens, 81 Staph. aureus, 2 P. aeruginosa, 35 Staphylococcus 

spp, 2 Strep. pneumoniae, and 2 E. faecalis were selected for testing. 

Isolates collected in the United States were selected from Focus Bio- 

Inova's (Hemdon, VA, USA) culture collection. European isolates were 

selected from the Intervet (Schwabenheim, Germany) culture 

collection.The results of the MIC determination and the size of 

inhibition zone for 30 g cefquinome disks were correlated and analyzed 

for error rates according to the CLSI M37-A2 standard in view of a set 

of MIC and disk diffusion zone breakpoints (susceptible < 2 g/ml or > 

22 mm; intermediate 4 g/ml or 19-22 mm; resistant 8 g/ml or < 18mm). 

Results: With the 30 g cefquinome disk the following error rate 

bounding occurred: no major error (falsely indicated susceptible by the 

disk) and only 1.3% (n=9) major errors (falsely indicated resistant by 

the disk) when MICs of £ 2 g/ml were interpreted as susceptible, 4 g/ml 

were interpreted as intermediate, and > 8 g/ml were interpreted as 

resistant for the broth micro-dilution and with an inhibition zone of < 

22 mm were interpreted as susceptible, 19-21 mm were interpreted as 

intermediate, and < 18 mm were interpreted as resistant for the disk 

diffusion zone diameters. 

Conclusion: Based on this study, 30 g cefquinome was determined to 

be a suitable disk load for use in cefquinome disk susceptibility testing. 

1139 Prevalence of Haemophilus somnus in Breeding Bulls: 

Bacteriological and Immunological Methods 

B. Vidic, S. Savic-Jevdjenic, Z. Grgic, I. Stojanov, D. Milanov, 

T. Barna 

Scientific Veterinary Institute Novi Sad, Laboratory of Microbiology 

and Infectious Diseases, Novi Sad, Serbia 

Introduction: H. somnus is etiological agent in several disease 

syndromes of cattle. In addition, H. somnus can be routinely isolated from 

mucus surfaces in apparently normal animal. Most bulls carry the 

organism asymptomatically in the prepuce and many cows are vaginal 

carriers. Isolation of H. somnus is possible only under specific conditions, 

also bacteria isolation is not a sure sign of pathogen influence. In detecting 

this disease serological methods would be of great help. 

Material and Method: As material a native bull sperm was used. For the 

cultivation of H. somnus, chocolate agar, Columbia agar, PPLO agar and 

brain-heart infusion agar were used. To nutritive media, the following was 

added: neomycin sulfate, nistatine, yeast extract and horse sera. The 

incubation was done at 37 °C, with the presence of 10% CO 2. Colonies 

which morphologically corresponded to H. somnus were further 

identified. The method of agglutination and m-CF test were used for 

detecting antibodies in blood sera from 93 breeder bulls. 

Results and Conclusion: H. somnus was isolated in two samples of 

bull sperm. After 24h on chocolate agar in the atmosphere with 10% 

CO 2, the same small colonies were noticed: 0.4-0.6mm large, rounded, 

with the smooth strait edges. After 48 and 72h, the colonies were 1,0- 

1,5 mm yellow pigmented, without shine, mildly granulated surface. 

After 48h on Columbia agar, the colonies were rounded, smooth, but 

after the prolonged incubation, the colonies became yellow, with 

mildly granulated surface. On nutritive agar, with the addition of horse 

serum, without blood, after 48h the colonies were rounded, shiny, 

smooth and lightly opalescent. The microorganism grew in the absence 

of both X and V factors. The microorganism was a pleomorphic Gram 

negative coccobacillus, catalasa negative, oxidase positive and it 

reduced nitrates and split aesculine. Sugar fermentation reactions, 48h 

after the inoculation were as follows: acid, but no gas was produced in 

dextrose, no fermentation of maltose, raffinose or rhamose. Out of 93 

examined bull blood sera, by the method of tube-agglutination (TA), 

the positive results were found in 2.15%, and by the method of m-CF in 

4.3% of the samples. The highest positive percentage was gained by the 

method of micro agglutination 9.68% and also the highest antibody 

titer was up to 1:128. Gained results suggest that there is a presence of 

the infection with H. somnus in breeding bulls. 

Key words: Haemophilus somnus, bulls, sperm 

1140 Flow Cytometric Analysis of Phagocytosis and Respiratory 

Burst in Leukocytes from Bovine Leukemia Virus Naturally 

Infected Cattle 

M. Azedo 1 , C. Massoco 2 , M. Blagitz 1 , B. Sanches 1 , F. Souza 1 , 

C. Batista 1 , U. Schalch 1 , F. Benesi 1 , A. Della Libera 1 

1 College of Veterinary Medicine and Animal Science of University of 

Sao Paulo, Department of Medical Clinics, Sâo Paulo, Brazil 

2 College of Veterinary Medicine of Cornell University, Department 

of Clinical Sciences, Ithaca, United States 

Bovine Leukemia Virus (BLV) is an oncogenic retrovirus that is 

associated with the development of persistent lymphocytosis (PL) and 

lymphoma in cattle. Assuming that events that lead to PL alters leukocyte 

subpopulations quantitatively and qualitatively, and influences the innate 

immune response, this study evaluated the intracellular production of 

hydrogen peroxide (H2O2) of circulating leukocytes after in vitro stimuli 

with phorbol-12-myristate-13-acetate (PMA), or Escherichia coli 

lipopolysaccharides (LPS), or after phagocytosis of propidium iodidelabeled 

Staphylococcus aureus (PI-Sa). Cells were obtained from 18 

BLV-negative cows, 16 naturally BLV-infected, non-lymphocytotic 

cows, and 15 BLV-positive cows with PL, and analyzed by flow 

cytometry. Mean percentage of H 2O 2-producing cells, characterized by 

the intensity of 2,7-dichlorofluorescein fluorescence, in the assay without 

in vitro stimulus, or after in vitro addition of PMA, or LPS, did not differ 

among groups. However, after in vitro phagocytosis of PI-Sa, the 

percentage of H 2O 2-producting leukocytes obtained from BLV-infected 

cows, presenting PL (34.06%) was smaller (p=0.002) than those verified 

in leukocytes obtained from non-infected (48.15%) and from BLVinfected, 

non-lymphocytotic, cows (44.87%). Furthermore, mean 

intracellular H 2O 2-production of leukocytes obtained from BLVinfected 

cows, presenting PL, without previous stimulus, or after the 

three different in vitro stimuli was smaller (p

The present work aimed to investigate nitric oxide (NO) and hydrogen 

peroxide (H 2O 2) production in monocyte-macrophage lineage from dairy 

cows naturally infected with bovine leukemia virus. Thus, 138 Holstein 

cows were casually sera tested by agar gel immunodiffusion (AGID) and 

the leukocyte profile was performed to discriminate the distinct groups 

known as alymphocytotic (AL) and persistent lymphocytosis (PL). The 

results on serologic test showed 45.65% positive dairy cows. From these 

positive animals 17.46% presented LP. From these cows, 30 animals were 

selected and divided equally in 3 groups (negative, AL and LP). The 

mononuclear cells from peripheral blood (PBMC) were isolated by 

gradient density centrifugation. The PBMC were incubated overnight and 

then the adherent cells were removed. Thus, 2x10 6 cells/ml monocytemacrophage 

lineage were plated per well for NO and H 2O 2 determination. 

NO synthesis in cell cultures was measured by Griess method and the 

H 2O 2 were analyzed using a simple colorimetric method with and without 

phorbol 12-myristate, 13-acetate (PMA). The median of H 2O 2 production 

from LP (28.98 nmol/mL) (p= 0.001) animals were higher than those 

observed from negative (SN) (6.74 nmol/mL) and AL (9.49 nmol/mL). 

The same were seen in H 2O 2 values with PMA stimulus (p

Material and Methods: The authors examinated post mortem 3372 

bovine in the 356 herds from January 2001 to December 2007 and 

collected informations from farmers and veterinarians. The study 

population included 25838 cattle, The main causes of deaths were 

classificated in eight categories: respiratory disorders,digestive 

disorders,calving disorders,metabolic disorders,accidents,udder/teat 

disorders,other known reasons and unknown reason. Data collected 

was analysed and compared. 

Results: Table 1 report the mortality rate in years from 2001 to 2007. 

Table 1. Deaths cattle and mortality rate 

2001 2002 2003 2004 2005 2006 2007 

N° deaths 496 492 447 446 498 491 502 

mortality 1,91% 1,90% 1,73% 1,73% 1,92% 1,90% 1,94% 

rate 

We found a higher mortality in older dairy cows (2007). The mortality 

rate rise from 3,41% in dairy cows 3-5 years old to 3,84% in dairy cows 

5-8 years old and 4,34% in dairy cows than 8 years older.In the beef 

breeds ( Piemontese ) the mortality rate is higher in cows 3-5 years old 

(1,63%) and cows 8 years older ( 1,67%) than cows 5-8 years old ( 

1,29%).The table 2 report the percentages to each group of death causes 

on dairy cows and beef breed cows. 

Table 2. Causes of death on dairy cow and beef breed cow mortality (%). 

Respi- Calving Meta- Accid- Udder Diges- Other Unratory 

dis- bolic ents /teat tive cause known 

disor- order disor- disor- disor causes 

ders ders ders ders 

Dairy 6 5 24 7 13 7 10 28 

cows 

Beef 7 25 4 3 2 10 18 31 

breeds 

cows 

The inter-herd variability was significant, with mortality rates running 

between 0 and 8%.We found that 44% of dairy cow deaths occurred 

within the first four weeeks of the lactation.All cattle twenty four 

months older (1472 deaths) were tested for Bovine Spongiform 

Encephalopathy (BSE) and were negative. 

Conclusions: We don’t found significant difference between the 

mortality rate in small and large herds. The quality of management 

result the major discriminant to reduce deaths cattle rate.A good 

practice breeeding has a strong positive impact on animal health, 

welfare and mortality. 

Key words: cows, cattle, mortality rate 

1146 Effect of Bovine Respiratory Disease on Growth and Carcass 

Quality during Early and Late Finishing Periods Using 

Treatment Records and Lung Lesions Scoring in French 

Feedlot Cattle 

F. Corbiere 1 , C. Lacroux 1 , R. Besse 1 , R. Chevalier 2 , PR. Cloet 2 , 

F. Schelcher 1 

1 



2 

Schering Plough Vétérinaire, Animal Health Division, Levallois 

Perret, France 

Aims: Evaluate the effect of bovine respiratory disease (BRD) and 

lung lesions on growth and carcass quality during early and late 

finishing periods in French feedlot cattle. 

Material and Methods: 407 Blonde d’Aquitaine five to seven months old 

male beef calves from a selection unit were monitored. Weight was 

recorded at entry (day one) and monthly until slaughtering after an average 

of 240 days on feed. The rectal temperature and clinical signs of BRD were 

recorded daily during the first 3 weeks. Onward, the rectal temperature was 

recorded only for calves with general or respiratory clinical signs. A 

treatment for BRD was initiated if rectal temperature was >40°C. At 

slaughter, the occurrence and extent of lung lesions were recorded and 

scored using a standardized scale and carcass quality was evaluated 

according to the EUROP scale. Associations between dependant variables 

(mean daily weight gain (ADG) and EUROP score) and the presence and 

extent of lung lesions or treatment for BRD were analyzed using linear 

mixed regression models adjusted for potential confounders. 

Results: Clinical BRD occurred in 52.8% of calves, mostly (77.6 %) 

during the first month on feed but mortality due to BRD remained low 

(0.48%). At slaughter, lung lesions were observed in 46.9% of calves, 

mostly in cranial pulmonary lobes. The prevalence of lung lesions was 

not significantly different between animals never treated for BRD, 

animals treated once and animals treated twice or more (p=0.53). The 

overall ADG was 1553g ± 155g. Treatment for BRD was associated 

with a 177g reduction in ADG (p



The Hazard Analysis and Critical Control Points (HACCP) system is a 

systematic approach that identifies, evaluates and controls steps in a 

process. By incorporating HACCP in a herd management setting, risk 

assessment, risk management and risk monitoring can be combined into 

one. In this pilot study, a HACCP-like approach was used to develop and 

apply management systems for mastitis control in dairy herds in Ireland. 

These systems were used to monitor the performance and effectiveness of 

control, and to guide the development of a focused and effective control 

response. Action research methodology was utilised, in conjunction with 

sociological and technical assessments, to inform the development and 

implementation of the HACCP-like approach. Six dairy farms were 

visited on four occasions over a twelve month period. An initial visit was 

conducted to gather farm data and explain the project to the farmer. 

Relevant hazards were identified and associated risks determined, 

allowing selection of critical control points and critical management 

points. Implementation of the HACCP strategy commenced after a second 

educational visit. Progress was monitored during two subsequent farm 

visits. The HACCP-like approach implemented used a holistic system of 

“farming best practice” and “generic critical control points” to monitor 

mastitis on all study farms. In conjunction, - farm specific critical control 

points and responses - were instigated as required. A monitoring system 

for the generic and specific critical control points was adapted for use on 

each farm. The action research component of the study draws on the 

principles of qualitative research allowing the project team to both 

evaluate and react to sociological and physical barriers to implementation, 

including farmers perceptions and expectations in the context of the 

HACCP program itself and in relation to interaction and cooperation with 

the team veterinarian. 

Key words: HACCP, mastitis, action research 

1149 Zinc Supplementation Effect in the Development and 

Conglutinin Concentration in Nelore Calves Thymus Raised 

in Brachiaria Decumbens in the Cerrado of Brazil 

S. Pereira 1 , C. Madruga 2 , S. Moraes 1 , I. Tozetti 3 , C. Faria 3 , 

A. Freitas 1 

1 

Embrapa, Gado de Corte, Campo Grande, Brazil 

2 

Escola de Medicina Veterinária UFBA, Gado de Corte, Salvador, 

Brazil 

3 

Universidade Federal do Mato Grosso do Sul, Campo Grande, 

Brazil 

Pregnant cows raised under extensive ranching conditions in Brachiaria 

decumbens pastures constituted two experimental groups. One group 

TZn0, the cows had administration of a full mineral mix, except zinc (Zn) 

and the other group TZn60 the same mineral mix, with 60 mg/kg of Zn. 

After two years of experiment the Zn intake by group TZn60 was 

significantly higher (X = 40 mg/day). The examination of ten thymus 

surgically excised from newborn calves of each experimental group 

evidenced in the morphometry that, thymus medullary and cortical zones 

were significantly larger in the TZn60 compared to TZn0 group (P < 0.05 

and P < 0.001, respectively). In the immunohistochemical test developed 

with CB1/8C1 monoclonal antibody, which recognizes epitope B in cattle 

native conglutinin, in recombinant conglutinin, and in the synthetic 

peptide corresponding to the 241-260 aminoacid sequence in the 

conglutinin molecule also the thymus of TZn60 showed more 

concentration of this C-type collectin, although the difference was not 

highly significant (P < 0.055). The results showed that Zn 

supplementation is essential for pregnant cows raised in pastures of 

tropical areas of world like Brazilian cerrado, where the soil is poor in Zn. 

This work showed that inorganic Zn supplementation through mineral 

mix influenced thymus development and conglutinin concentration in this 

primary lymphoid organ. Probably these calves have better 

immunological structure to respond more effectively to pathogenic 

agents, particularly in young calves. Probably, the conglutinin plays a 

relevant role in the biology of bovine thymus, however elucidation of the 

mechanisms involved warrants further research. 

Key words: cattle, thymus, zinc, conglutinin, immunohistochemistry 

1150 Association between Subclinical Endometritis and Glycogen 

Concentration in Neutrophils of Lactating Holstein Cows 

K. Galvao, W. Butler, S. Brittin, L. Caixeta, R. Sper, M. Fraga, 

A. Ricci, R. Gilbert 


Cornell University, Ithaca, United States 

Neutrophils (PMN) depend on anaerobic glycolysis for the energy 

required for chemotaxis, phagocytosis, and microbial killing; therefore, 

periods of negative energy balance may deplete glucose stores and 

affect PMN function. The objective was to measure glycogen stored in 

PMN and to evaluate its association with subclinical endometritis (SE) 

in dairy cows. Holstein cows (51), had 60 mL of blood collected 

weekly for PMN isolation from calving until 21 d in milk (DIM) then 

had another sample collected at 35 DIM and had a uterine lavage for 

cytology at 42 DIM. Ten million PMN were isolated and assayed for 

glycogen quantification. Cows that developed postpartum metritis were 

not included in the study. Subclinical endometritis was characterized 

by the presence of 5% or more PMN in the uterine cytology and 45% 

of the cows were classified as having SE. Body condition score (BCS) 

and blood glucose concentration were also evaluated at sampling. 

Association between SE, glycogen, and glucose concentration was 

evaluated using the MIXED procedure of SAS for repeated measures; 

the model also included the effects of BCS, DIM at sampling and 

interaction between SE and DIM at sampling. There was no effect of 

SE, glucose, or BCS on PMN glycogen concentration; however, there 

was an effect of DIM at sampling (P < 0.001) and an interaction 

between SE and DIM at sampling (P = 0.01). The PMN glycogen 

concentration was 33.7, 26.1, 21.5, 22.3, and 24.1 µg glycogen/10 6 

PMN at 0, 7, 14, 21, and 35 DIM, and was greater (P < 0.05) at 0 than 

the other time points and at 7 than at 14 DIM. Cows with subclinical 

endometritis had lower PMN glycogen concentration only at 7 DIM 

(22.5 vs 29.7; P = 0.03). Subclinical endometritis was not associated 

with glucose concentration; however, DIM at sampling, and parity 

were. Glucose was 73.6, 62.5, 63.1, 64.7 and 65.8 mg/dL at 0, 7, 14, 21, 

and 35 DIM and was greater (P < 0.001) at 0 than the other days. 

Multiparous cows had lower glucose concentration (62.7 vs 69.1;P < 

0.001). We conclude from this study that subclinical endometritis was 

not associated with PMN glycogen concentration over all time points. 

Nevertheless, an interaction with DIM was observed and PMN 

glycogen concentration was lower in cows with SE at 7 DIM. Although 

PMN glycogen and glucose did not show a direct association, they 

followed a similar pattern after calving, indicating that PMN glycogen 

concentration may be affected by the cows’ metabolic state. 

1151 Efficacy of Vaccination against Ringworm in Veal Calves - 

Multicenter Controlled Field Study 

P. Houffschmitt 1 , Y. Lagalisse 1 , P. Personne 2 

1 

Intervet Schering-Plough, Technical Services, Angers, France 

2 

Syndicat National des Cuirs et Peaux, Technical Services, Paris, 

France 

Objectives: The objective of the study was to evaluate the efficacy of a 

single dose vaccination in veal calves with a live attenuated 

Trichophyton verrucosum strain LTF-130 on the day of arrival in their 

fattening units, as a preventive tool against both skin lesions and leather 

damage caused by infection with this zoophilic fungus. 

Materials and methods: The study was performed with veal calves 

housed in standard fattening units. At arrival to the unit the animals 

were randomly allocated to a vaccinated or non vaccinated control 

group. The vaccinated animals received a single dose of a live 

attenuated ringworm vaccine (Bovilis ® Ringvac, Intervet). In a first 

series of trials (1, 2, 3a), skin lesions were assessed and monitored by 

veterinarians for up to 11 weeks after arrival (Day 0). In a second series 

of trials (3b, 4), evaluation of vaccine efficacy was based on evaluation 

of the skins during post slaughter processing. The hides were then 

carefully examined by a specialist, as wet blue hides, and given quality 

scores (perfect/ damaged/ very poor). 

Results: In the first series of trials vaccinated calves showed significantly 

fewer skin lesions than non vaccinated controls. The number of 

vaccinated calves with no lesions varied from 87% to 99%, as compared 

to 47% to 82% in the unvaccinated calves. The maximum benefit was 

observed in trial 1, where only 1 % of the vaccinated animals showed skin 

lesions (18 % in the control group, p< 0.01). In the second series of trials 

hides from vaccinated calves were given better quality scores after 

staining than hides from unvaccinated calves (p< 0.01). 

Conclusions: A single dose vaccination with the live attenuated T. 

verrucosum strain LTF-130 (Bovilis ® Ringvac) of veal calves 

performed on the day of arrival in the fattening unit, proves to be an 

efficient preventive tool against ringworm clinical signs and its 

negative influence on hide quality.

211 Endocrine Status of Holstein Dairy Cows With Different Milk 

Production during late Pregnancy and early Lactation 

D. Gvozdic 1 , H. Samanc 2 , B. Kovacevic 4 , D. Kirovski 3 , N. Fratric 3 , 

I. Vujanac 2 , B. Dimitrijevic 2 

1 


pathophysiology, Belgrade, Serbia 

2 

Faculty of Veterinary Medicine Belgrade, Large Animal Clinic, 


3 


Physiology, Belgrade, Serbia 

4 

Veterinary station, Veterinary Department, Teslic, Bosnia and 

Herzegovina 

Aim of this work was to determine endocrine status of Holstein 

dairy cows with different milk production in various stages of 

reproductive cycle. Total number of 60 Holstein dairy cows has 

been selected during late pregnancy on the basis of average milk 

production during the last lactation and divided in two groups: 1) 

high production group (HP) - cows with average milk production 

exceeding 7500L (n=30), 2) low production group (LP) - cows with 

average milk production lower than 6500L (n=30). Blood serum 

thyroxine (T4), triiodothyronine (T3), and insulin (Ins) 

concentrations were determined in blood samples obtained from v. 

jugularis during the last week prior to partus, between 7-14 days 

after partus, and two months after beginning of lactation.Blood 

serum thyroid hormones concentration was similar in both groups of 

cows during the late pregnancy, decreasing after partus and two 

months after beginning of lactation. Blood serum T4 and T3 

concentrations were significantly lower in HP cows after partus and 

two months after beginning of lactation (33.41±6.48:27.77±5.22 

and 32.76±4.44:25.19±5.36 nmol/L, p

214 Cystic Ovarian Diseases in Dairy Cows: Epidural GnRH and 

Calcium/Naloxone Administration 

M. Cinone 1 , G. Minoia 1 , D. Robbe 1 , A. Guaricci 1 , D. Coronati 3 , 

M. Mutinati 1 , A. Rizzo 1 

1 Faculty of Veterinary Medicine of Bari, Department of Animal 

Production, Valenzano (BA), Italy 

2 University of Teramo, Clinical Veterinary Scieces, teramo, Italy 

3 CIRIO Agricola Farm Animals, Veterinarian, Caserta, Italy 

Objectives: Cystic ovarian diseases (COD) occur in postpartum 

dairy cows and are characterized by the presence of anovulatory 

ovarian structures. Their etiopathogenesis is multifactorial and 

related to the asynchrony in the induction of LH receptors on the 

granulosa and theca cells during follicular maturation or to their 

desensitization. Endogenous opioid peptides (EOP) have been 

documented in several parts of the reproductive tract and are 

involved in the control of the stress-induced changes in LH 

secretion. The possibility of an intragonadal local opioidergic 

control cannot be excluded because the presence of ‚-endorphins (‚end) 

has been detected in the ovary of some mammals. The present 

study was designed 1) to measure -end concentrations in bovine 

cystic fluid; 2) to evaluate the impact of the epidural administration 

of GnRH and calcium/naloxone (Ca/Nx) on EOP removal. 

Materials and methods: The experimental study was performed on 

45 Friesian cows (60 d. postpartum) affected by COD and on 5 

oestrus cows. Animals were assigned to 4 groups: A- 30 cows 

undergoing an epidural treatment with 3ml of Ca/Nx (naloxone 

hydrochloride 0.6mg dissolved in 20% calcium gluconate) and 2ml 

of GnRH (25ng/ml lecirelina ® ), twice, in an interval of 21 d; B- 10 

cows receiving 5ml of 0.9% NaCl solution, epidural via; C- 5 cows 

in which cystic fluid was aspired in vivo; D- 5 oestrus cows in which 

the fluid from the preovulatory dominant follicle was aspired. 

Oestrus occurred in all cows, thus AI was performed. Pregnancy was 

confirmed 30 d. after AI. 15/30 cows with COD showed oestrus in 

the following 18±3 d. after GnRH+Ca/Nx treatment. Conception rate 

was 66% (10/15 cows). Plasma ‚-end concentration increased 

(1.9±1ng/ml) after the 1st administration and decreased up to 

0.5±0.2ng/ml (p

lactating Holstein Friesian cows and six Japanese Black beef cows 

had been ovariectomized before the experiment. The devices were 

inserted into the vagina 2 to 3 weeks after the ovariectomy and 

removed 12 days after the insertion. Blood was collected via the tail 

venepuncture once a day at 8.00 a.m. from the day before insertion 

to 15 days after the insertion. On the day of insertion plasma samples 

were also collected at 0, 1, 2, 4, 6, and 8 h after the insertion. Plasma 

P4 concentrations were determined by ELISA. The experiment was 

repeated in some animals at 8 to 10 d intervals. The plasma P4 

concentrations in cows of the DIB group (n=7) increased to 2.6 ± 1.0 

ng/ml at 1h, and to 4.6 ± 1.4 ng/ml at 2 h and, thereafter, remained 

above 1.0 ng/ml until the 12th day. Likewise, plasma P4 

concentrations in cow of CIDR group (n=7) rose to 1.6 ± 0.6 ng/ml 

at 1h, and to 2.7 ± 1.3 ng/ml at 2 h after the insertion. A high P4 

concentration above 1.0 ng/ml was maintained until the day of 

removal. There was no significant difference in the plasma P4 

profiles between the dairy and beef cows. Thus, the new intravaginal 

device is shown to be effective to increase plasma P4 

concentrations within a few hours after insertion and to maintain the 

elevated levels during a treatment period of 12 d in cows, which is 

well comparable with the effectiveness of CIDR. 

Key words: DIB, CIDR, progesterone device, Holstein Friesian, 

Japanese Black 

255 Control of Oestrus with a Progesterone Intravaginal Device 

(Prid ® ): Comparison of 2 Insertion Durations 

S. Floc’H 1 , F. Deletang 2 , S. Freret 3 , C. Ponsart 4 , D. Remmy 5 

1 CEVA santé Animale, Livestock Business Unit, Libourne, France 

2 Animal Reproduction, La Rochelle, France 

3 INRA, Animal Reproduction, Tours, France, France 

4 UNCEIA, R&D, Maisons-Alfort, France 

5 CEVA Santé Animale, Ruminants, Libourne, France 

The objective of this study was to optimise the efficacy of PRID by 

comparing 2 insertion durations, 7 and 9 days, in term of pregnancy 

rate after a single fixed time artificial insemination. This study was 

conducted during 2 years: firstly in beef cattle (n=748) and 

secondly in dairy ccattle (n=498). In beef cattle only, blood was 

sampled twice before treatment for determination of plasma 

progesterone to check cyclicity: 66.2 % of females were defined as 

cyclic at the beginning of the treatment. Pregnancy was checked by 

transrectal ultrasonography 35 days post insemination or by 

pregnancy Specific Protein B (PSPB) dosage by RIA 45 days post 

insemination or by rectal palpation. Treatment protocol was : PRID 

insertion for 7 or 9 days, PgF2 injection 24h before removal of 

PRID, equine Chorionic Gonadotropin (eCG) was injected at the 

time of removal : from 400 to 600 IU according to breed and 

physiological status. Cows were bred once 56h after PRID removal. 

The global pregnancy rate was 60.1 % (n=749/1246). The treatment 

duration influenced this pregnancy rate as it tended to be higher (p = 

0.056) in the 9 days group (62.8 %) vs. 7 days group (57.5 %). The 

effect of calving rank was observed with a higher pregnancy rate in 

heifers (65 %) than in cows (53.2 %). To compare the 2 groups, 

higher differences in pregnancy rates in the 9 vs. 7 days group were 

observed in heifers in beef cattle (78.5 % vs. 68.5 %) and in cows in 

dairy cattle (42.9 % vs. 33.3 %). The effect of body condition score 

(BCS) was observed only in dairy cows with a tendency (p = 0.09) 

for higher pregnancy rate in animals with a BCS of 3 vs. those with 

a BCS 2.5 or 3.5 (54.7 % vs. 45 % respectively). Further, higher 

pregnancy rates were observed in the 9 vs. 7 days group in dairy 

cows with BCS 2.5 (50 % vs. 41.9 %). Regarding cyclicity in beef 

cattle, pregnancy rate was significantly higher (p = 0.0002) in 

cycling (71.2 %) vs. non cycling animals (57.4 %). The highest 

difference between 9 and 7 days group was observed in non cycling 

females (64.3 % and 50 % resp.). Results of this study showed better 

pregnancy rates with a 9 vs. 7 days insertion of PRID (62.8 % vs. 

57.5 % respectively), it was then concluded that a 9 days insertion 

tends to improve fertility (p = 0.056), more particularly in non 

cycling vs. cycling animals and in dairy cattle in cows vs. heifers and 

in animals with BCS < 3. 

Key words: progesterone, PRID, treatment duration 

256 Effect of Progesterone Supplementation post Conception on 

Embryo Survival Rate in Heifers 

M. Beltman 1 , P. Lonergan 2 , J. Roche 2 , M. Diskin 3 , M. Crowe 2 

1 

University College Dublin, Unit of Herd and Veterinary Public 

Health, Dublin, Ireland 

2 

University College Dublin, Animal and Veterinary Science, Dublin, 


3 

Teagasc, Animal Production Research Centre, Athenry, Ireland 

Progesterone (P4) is essential for establishment and maintenance of 

pregnancy in mammals. The objective of this study was to examine 

the effect of elevating P4 during the first week post conception on 

embryo survival. Oestrus was synchronized in beef cross heifers 

(n=197, ~2-years old) using two injections of prostaglandin F 2a (PG, 

Prosolvin, Intervet Irl.) 9 days apart. The heifers were checked for 

oestrus 4x daily commencing 24h after the second PG and were 

inseminated 12-18 h after oestrus onset (=day 0). Inseminated 

heifers were randomly assigned to 1 of 3 groups: (i) Control, n=69; 

(ii) P4 supplementation using a Controlled Internal Drug Device 

(CIDR 1.9g, Pfizer UK) from days 3 to 6.5, n=64; or (iii) P4 (CIDR) 

from days 4.5 to 8, n=64. Body condition (BCS) and locomotion 

scores (scale of 1-5) were recorded for all animals. Animals with a 

locomotion score 4 (very lame) were excluded. Pregnancy was 

determined at slaughter on day 25. Conceptus length and weight 

were recorded and the corpora lutea (CL) of all pregnant animals 

were weighed. Embryo survival rates were compared by Chi-square 

test and the relationships between BCS, locomotion score, P4 levels 

and embryo survival rates were analysed using regression analysis. 

Supplementation with exogenous P4 increased (P

inseminator were tested in regression models and terms were 

retained in the model if influential. Five days after treatment, the 

proportion of cattle with more than one CL was 72% and 7% 

(P11,000 kg of 

milk/cow/year, and use automatic pedometry systems (recording 

cow steps) for heat detection. A controlled randomized trial was 

conducted from July 2006 until December 2007. Anoestrus cows 

were treated according to each farm protocols. Total of 1450 cows of 

all lactations after artificial insemination (AI) were randomly 

allocated into tow nearly equal groups: US and RP. The RP cows 

were submitted to PD by fetal membrane slip technique between 

days 42-48 after AI, and the US cows were PD between days 26-32 

after AI and reexamined by RP between days 42-48 after AI. Cows 

which were detected on heat between AI and the set PD time in each 

group were reinseminated. Results: Days open were 122 and 123 in 

herd B and 115 and 115 in herd A, in the US and in the RP groups 

respectively, with no statistically significant differences. The overall 

conception rate from first insemination was 42.7 for herd A and 44.3 

for herd B. The groups did not differ significantly in following 

reproductive risk factors: endometritis, ketosis, negative energy 

balance and summer calving. Most of the active open cows were 

detected on heat and reinseminated before reaching the first PD. The 

EED rate in the US group, measured by the drop of pregnancy rate 

between the day 26-32 and the day 42-48 PD, was 5.9% and 6.1% in 

farm A and B respectively. 

Conclusion: There was no difference in days open and therefore no 

reproductive benefit in early ultrasound pregnancy diagnosis in these 

cows herds. As part of the open cows were detected on heat before the 

US PD and the true anestrous cows take longer to conceive, the 

potential 16 days advantage in days open for the US cows was reduced 

to an insignificant difference. 

303 Resynchronization and Early Pregnancy Detection may 

Enhance Reproductive Performance of Lactating Dairy Cows 

A. Niasari-Naslaji 1 , M. Bolourchi 1 , S. Shabani 2 , D. Nikjou 1 , 

A. Moghiseh 1 



2 Bosatn Agro-Inductrial Co, Hashtgerd, Iran 

Early identification of non-pregnant cows using ultrasonography and 

their resynchronization using GnRH-PG protocol has been applied 

recently as a managerial tool to enhance reproductive performance 

of lactating dairy cows. During 8 months study in a well-managed 

dairy farm, 141 healthy lactating dairy cows, without any particular 

reproductive problems, >45 days open, were selected, randomly 

assigned into two group at the time of AI and monitored until they 

became pregnant. Experimental cows were inseminated 12 hrs after 

observing standing estrus using frozen semen. Cows in control group 

(n=67) did not receive any treatment. On Day 20-25 after AI, cows 

in treatment group were scanned ultrasonically (Sonovet 2000, 5-9 


MHz linear rectal transducer) for the characterization of follicle and 

CL on the ovary and given an ovulating dose of GnRH (15 µg 

Luliberin-A, Vetaroline, Aburaraihan, Iran). On Day 27-32 after AI, 

pregnancy was diagnosed using ultrasonography and non-pregnant 

cows received prostaglandin F2 analogue (PG, 500 µg cloprostenol). 

Cows that returned to estrus were re-inseminated 12 hrs after 

observing standing estrus. Pregnancy was rechecked on Day 50-60 

days after AI by rectal examination. Data with discrete nature were 

analyzed using Genmod procedure including logit in the model and 

data with continuous nature were subjected to GLM procedure 

following appropriate transformation. On Day 20-25 after AI, 

ovarian follicle (16.8±0.42; 10-30 mm) and CL (27.8±0.45; 15-35 

mm) were recorded for resynchronized cows. The number of days 

open, the days from AI, followed by resynchronization, to pregnancy 

and total services per conception were significantly higher (P

a better indicator for progesterone synthesis of the luteal gland in 

cyclic cows. For this reason luteal size (LS), luteal blood flow (LBF) 

and plasma progesterone level (P4) were examined during the 

estrous cycle in 14 cyclic Holstein Friesian cows. Examinations 

were carried out on Days 3, 6, 9, 11, 13, 15 and 18 of the estrous 

cycle (0 = day of estrus). LS was quantified by measuring the 

maximum cross-sectional area of the CL on a digitized B-mode 

image and P 4 was determined by EIA. LBF was investigated using 

Power Doppler Mode. The maximum area of colour pixels within the 

Corpus luteum (CL) represented luteal blood flow as a semiquantitative 

parameter for luteal perfusion. During the estrous cycle 

similar patterns of changes in LS, LBF and P 4 occurred (r > 0.90; P 

< 0.0001). The parameters increased between Days 3 and 9 (P 4) and 

11 (LS, LBF), respectively, stayed at constant levels until Days 13 

(LS) and 15 (LBF, P 4), respectively. LS decreased (P

University of Nottingham, Animal Health and Welfare, Nottingham, 


In this presentation the author would like to stimulate thoughts 

regarding the choice of performing a caesarean or fetotomy when 

dealing with a dead foetus during bovine parturition. A review of the 

literature present on this topic will be summarized to show scientific 

evidence on which decision-making in particular situations can be 

based. What is known about the prognosis of an animal after either 

procedure, do we know how each procedure influences future 

performance? The current situation suggests that veterinarians steer 

away from performing fetotomies and prefer to perform a caesarean, 

for a variety of reasons. However, anecdotal evidence suggests these 

reasons may not necessarily be based on correct clinical evaluation 

of the situation but on experience of the veterinarian or use of 

adequate equipment. With veterinarians becoming more specialised 

within their profession the question is raised whether it is important 

for bovine veterinarians to be thoroughly trained to perform both 

procedures adequately. Advantages and disadvantages for both 

techniques will be presented and experiences from the public will be 

sought to stimulate discussion. 

Key words: obstetrics, Caesarean section, fetotomy, dystocia, foetus 

372 Comparison of the Changes in Intrauterine Pressure after 

Treatment with Various Prostaglandins in Early Postpartum 

Cows 

Á. Bajcsy 1 , O. Szenci 1 , G. Van der Weijden 2 , A. Doornenbal 2 , 

J. Bartyik 3 , F. Rezazadeh 1 , K. Szabó-Ari 1 , M. Taverne 2 


Large Animals, Üll[, Hungary 

2 Faculty of Veterinary Medicine, Utrecht University, Department of 

Farm Animal Health, Utrecht, The Netherlands 

3 Enying Agricultural Ltd., Kiscséripuszta, Hungary 

Our previous study in postpartum (pp.) cows without retained 

placenta showed that a single treatment with either oxytocin or 

carbetocin (administered between 14h and 16h pp.) significantly 

increased both contraction frequency (FREQ) and total area under 

the curve (TAUC) during the 1 st post-treatment hour, with a return to 

initial levels during the 3 rd hour (2006,Theriogenology 65: 486-498). 

Because F series prostaglandins are known to stimulate uterine 

contractility in non-pregnant cycling cows, the present study was 

performed to quantify possible uterotonic effects of therapeutical 

(=luteolytic) doses of natural PGF 2a and two synthetic analogues of 

PGF 2a in early pp. cows. 

Thirty nine pluriparous Holstein-Friesian cows were included in a farm 

study. All cows had expelled their fetal membranes within 12h after 

uncomplicated calving. They were randomly attributed to 1 of 4 treatment 

groups (all treatments given as a single im. injection between 14h and 17h 

pp.): Group 1 (n=10): 25mg natural PGF 2a (Enzaprost ® ); Group 2 (n=9): 

500 µg cloprostenol (Estrumate ® ); Group 3 (n=10): 150 µg Dcloprostenol 

(Genestran ® ); Group 4 (n=10): 5ml saline solution. For 

recording intrauterine pressure (IUP), an open tip catheter was 

transcervically introduced into the previously pregnant uterine horn prior 

to treatment and was fixed to a caruncle. The acquired signals were 

digitalized (at 4Hz) and analyzed, using a graphical programming 

software (Labview 5.0). The IUP characteristics during each of the first 

4h, and the 12th and 24th h after treatment were evaluated among the 

groups, using RM MANOVA. 

The treatments did not significantly affect any of the following IUP 

parameters: FREQ, amplitude (AMP), area under the curve (AUC) and 

TAUC. However, duration (DUR) slightly differed among groups 

(P

clear intraluminal mucus, and in 5 cases purulent endometritis was 

found. In contrast, only two uteri were without any histopathological 

findings. Histological alterations, varying in quantity and quality, 

could be detected in 116 uteri: 101 cows showed a periglandular 

fibrosis (“bovine endometrosis”), 84 angiopathies, and 72 an 

endometritis, mostly nonpurulent. In most cases (n=104), several 

alterations occurred simultaneously. Due to sampling artefacts, only 

77 of 98 biopsies could be examined histologically. In 48 of 49 

cases, at least one of both biopsies was evaluable. There is a farranging 

conformity comparing the findings in biopsy with those in 

uterine samples, collected post mortem. These results indicate that 

the majority of sub-/infertile cows without further clinical symptoms 

has endometrial alterations visible in endometrial biopsy. 

Nevertheless, in order to establish the potential diagnostic tool 

“endometrial biopsy” in buiatrics as a prognostically meaningful 

method, further investigations, especially in clinically healthy and 

fertile animals with a detailed documented reproductive status, are 

necessary. 

Key words: cattle, reproduction, endometrial biopsy, infertility, uterus 

375 The Effect of Buserelin and an Opioid Antagonist on 

Pregnancy Rate of Repeat Breeding Holstein Dairy Cows 

V. Fuentes, A. Bernal, J. Olmedo 

Universidad de Guadalajara, Centro Universitario de los Altos, 

Tepatitlan Jalisco Mexico, Mexico 

The objective of this study was to observe the effect of buserelin and 

naloxone in repeat breeder Holstein dairy cows. In a double blind 

study 45 repeat breeder Holstein cows were selected from a large 

dairy cooperative farm of the highlands of the occident of Mexico. 

All cows presented a history of repeat breeding, and further more, 

during palpation the presence of an ovaryan cyst was detected. 

Divided at random in three groups of 15 cows. Group one was 

treated with 15 µg buserelin im on the oestrus day of insemination. 

Group 2 was treated with naloxone, 5 mg im at 12 hour intervals for 

three consecutive days, treatment commenced since one day before 

insemination. Group 3 was used as a control injected with 5 ml im of 

saline solution at 12 hour intervals for three consecutive days, 

injections commenced since one day before estrual insemination. 

Cows were inseminated twice (12 h apart) according to the AM-PM 

rule and pregnancy was confirmed by rectal palpation between 12 

and 18 weecks after the last insemination. Pregnancy rates in 

buserelin treated repeat breeder cows was 56%, while pregancy rate 

in naloxone treated cows was 89% and control cows showed a 20% 

pregancy rate. There was a significant correlation betweein 

progesterone levels and pregancy rate in all groups. It was concluded 

that opioids participate significantly in expression of estrous 

behaviour and ovulation in repeat breeder Holstein cows 

Key words: buserelin, naloxone, dairy cows 

376 The Incidence of Repeat Breeding and Subsequent 

Reproductive Performance in High-producing Dairy Cows 

T. Nakao 1 , Y. Mohammad 1 , K. Koike 1 , C. Yoshida 2 , K. Yamada 3 

1 Yamaguchi University, Department of Veterinary Medicine, 

Yamaguchi, Japan 

2 Niigata University, Facuty of Agriculture, Niigata, Japan 

3 Nemoro NOSAI, Shibetsu Large Animal Clinic, Shibetsu, Japan 

Declining fertility in lactating dairy cows has recently been reported 

worldwide. The underlining causes of the subfertility have been 

known to be the increase in milk yield and resultant negative energy 

balance during an early lactation period. It is assumed that, as the 

consequence of reduced fertility, the incidence of repeat breeding 

has increased in today’s high producing cows. This paper aims to 

show the incidence of repeat breeding in eight commercial dairy 

herds across Japan, which were monitored for ovarian cyclicity and 

reproductive performance by twice weekly milk progesterone assayd 

and regular herd visits during a period from April 2004 to July 2005. 

Of a total of 274 cows, 15.2% cows had not conceived after the third 

AI or the subsequent AI without showing clinical reproductive 

abnormalities (RB cows). The incidence of repeat breeding among 

eight different herds ranged between 6.8 and 20.3%. Two hundred 

and one cows conceived after first, second, or third AI (fertile cows). 

The RB cows, in comparison with the fertile cows, showed a lower 

pregnancy rate within 210 d after calving (38.9% vs. 

94.5%)(P

tissue (8,6 cm 2 ) was twice as large, the TW (3,2 mm) was three times 

as big and the CA (1,8 cm 2 ) was 4.5 times as big compared to 

follicles without luteal tissue. The sensitivity of diagnosing 

luteinized follicles was 61.5% using B-mode sonography and 92.3% 

using Doppler sonography. The specificity was 100% for both 

methods. Seven of the twelve cows were responding to treatment, 

showing an increase of P4 values above 1.0 ng/mL ten days after 

treatment, while in the other 5 cows P4 values stayed at values lower 

than 1.0 ng/mL. The blood flow in responding animals tended to 

increase (P=0.07) 30 minutes after treatment by 59.3%, while in the 

not responding cows no changes could be observed (P=0.50). In 

conclusion, this study demonstrates, transrectal Doppler sonography 

of follicular blood is not useful to predict the persistence of follicles, 

but it is a useful method to differentiate between follicle and luteal 

cysts. The evaluation of follicular blood flow shortly after GnRHinjection 

is not a reliable method for the success of treatment, but 

gives some indications about it. 

Key words: cystic ovarian follicles; Doppler, GnRH, cow 

379 Efficacy and Accuracy of three Oestrus Detection Techniques 

in a Confinement or Pasture Dairy Management System 

J. Mee 1 , M. Palmer 2 , G. Olmos 1 , L. Boyle 1 

1 



2 

University of Edinburgh, Royal (Dick) School of Veterinary Studies, 

Edinburgh, United Kingdom 

The objective of this experiment was to compare the efficacy of 

three methods of oestrus detection [visual observation (VO), tail 

paint (TP) and radiotelemetry-HeatWatch ® (HW)] in two 

management systems [cubicle housing with a total mixed ration 

(HOUSED) and rotational pasture with concentrate supplementation 

(GRASS)]. The 46 randomly allocated and blocked, spring-calving 

Holstein-Friesian cows were monitored by the three oestrus 

detection methods simultaneously from ten days postcalving for nine 

weeks on the same farm. The occurrence of nine selected behaviours 

associated with oestrus was also recorded during the thrice daily 20 

minute visual observation sessions. Thrice weekly milk sampling for 

progesterone analysis (EIA) was used to determine the dates of true 

standing oestrus events (oestrus detection accuracy). Data were 

analysed by proc Frequency, Genmod, Npar1way, Ttest and 

Univariate, as appropriate, in SAS. All three detection methods had 

a higher oestrus detection efficacy in the GRASS (VO 59, TP 65 and 

HW 69%) compared to the HOUSED treatment (VO 20, TP 26 and 

HW 37%) (P

the three herds showed a significant improvement on first service 

and overall conception rates (p

libido in unmotivated males. Similarly, identification of diestrus 

specific chemicals in urine may be applicable to monitor the 

cyclicity. The bull specific compounds are immense beneficial in 

induction of estrus in delayed pubertal or postpartum anestrus 

female and also to enhance estrus intensity in silent estrus 

individual. In the present study, urinary volatile chemical 

compounds were identified in samples from cows at estrus and 

diestrus stage and from sexually active bulls, using gas 

chromatography mass spectrometry (GC-MS). Dichloromethane 

was used for extraction of urine and identification of analytes 

compound was carried out in Perkin Elmer gas chromatograph 

linked to mass spectrometry. The analyses of cow and bull urine 

revealed the presence of total fifteen compounds with molecular 

weights ranging from 114 to 446 Da. GC profile of estrus cow urine 

was specified by the presence of two compounds viz., Octane and 3hexanol 

2,2-dimethyl, whereas, only one compound (decane, 6ethyl, 

2-methyl) was identified from urine of cows at diestrus stage. 

The chromatographic profile of bull urine was demarcated by the 

presence of three specific compounds namely, nonacosane; 1,2, 

benzene dicarboxylic acid- dihexyl ester and heptacosane. The 

present study is a novel attempt to provide valuable chemical 

information on the urinary compounds that might have potential role 

in olfactory communication among the conspecific members. 

However, behavioural assay with synthetic compounds is indicated 

to acquire more information on the olfactory communication and to 

identify the sex or estrus specific marker in this species. 

Key words: urine, chemo-signal, cow, bull, gas chromatography-mass 

spectrometry 

853 The Effect of Season and Parity on Heat-synch Program in 

Dairy Cows 

M. Bolourchi, R. Emadi, A. Niasari-Naslaji, P. Hovareshti 

Faculty of Veterinary Medicine, University of Tehran, Department of 

Clinical Sciences, Faculty of Veterinary Medicine, Tehran, Iran 

This study was conducted to investigate the effect of season and parity 

on Heat-Synch program in dairy cows. Dairy cows (n=230) from two 

dairy farms located in the vicinity of Tehran,with the average daily 

milk of 30 kg and with voluntary waiting period of 50 days were 

assigned into 2 groups (Control and Heat-Synch) during 4 seasons 

considering their parity (uniparous and multiparous). All cows received 

2 luteolytic doses of prostaglandin F 2a analogue (PGF; 500 µg 

cloprostenol, Nasr Company, Iran) on Days 35 and 48 after parturition. 

Cows in the control group (n=148) did not receive any further 

treatment and artificially inseminated with frozen semen 12 hours after 

showing standing estrus. Cows in Heat-Synch program (n=82) received 

an intramuscular injection of 12.5 µg GnRH analogue (Luliberin, 

Aburaihan, Iran; Day 0) followed by an intramuscular injection of PGF 

on Day 7. On Day 8, females in these groups received an intramuscular 

injection of estradiol benzoate (1 mg; Aburaihan, Iran). Estrous 

detection was performed for 48 hours after estradiol injection. Females, 

displayed standing estrus, were inseminated with frozen semen. Those 

females that did not display estrus were inseminated blindly at 48 hours 

after estradiol injection. Pregnancy was diagnosed on Day 35-40 after 

AI via rectal palpation. First service conception rates were analyzed 

using Genmod procedure in SAS including logit statement in the 

model. First Service conception rates were not different among groups 

between the two dairy farms. Therefore data from these dairy farms 

were pooled for further analyses. There was no significant difference 

between two groups regarding different seasons [Control: spring: 

39.53% (17/43); summer: 61.11% (11/18); fall: 57.45% (27/47); 

winter: 55% (22/40); Heat-Synch: spring: 64% (16/25); summer: 

66.67% (6/9); fall: 50% (11/22); winter: 57.69% (15/26); P>0.05]. 

There was also no significant difference between two groups regarding 

their parity [Control: uni-parous: 48.6% (18/37); multi-parous: 53.1% 

(59/111); Heat-Synch: uniparous: 35.7% (5/14); multiparous: 64.2% 

(43/67); P>0.05]. In conclusion, season and parity did not have any 

significant effects on first service conception rates in dairy cows 

enrolled in Heat-Synch program compared to control. 

854 Determination of PAG in the Milk and Blood in Cow’s in 

Comparison to other Examination Methods for Pregnancy 

Diagnosis 

Z. Gajewski 1 , N. Melo de Sousa 2 , A. A Konopka 1 , JF. Beckers 2 , 

R. Thun 3 


1 

Warsaw University of Life Sciences, Departament of Animal 

Reproduction, Warsaw, Poland 

2 

University of Liege, Department Physiology of Reproduction, Liege, 

Belgium 

3 

University of Zurich, Clinic of Reproduction, Zurich, Switzerland 

Introduction: In clinical practice a few pregnancy diagnostic methods, 

which differ in effectiveness and time are used, one of the methods is 

measuring PAG (Pregnancy Associated Glycoprotein) concentration in 

milk and blood. 

Aims and methods: the aim of the present study was to use a new 

developed method for evaluating the PAG in milk and blood and 

compare these results with USG and rectal investigation for 

pregnancy diagnosis in the cows. The experiment was carried out on 

a group of 220 cows of HF breed. The used animals were 3 to 8 years 

old. All animals underwent pregnancy testing: ultrasonography with 

an ALOKA SSD-500 with 5.0 MHz probe, 30-55 days after 

insemination and rectal control after the 55 th day of pregnancy. 

Blood and milk were used for PAG tests. The concentration of two 

PAGs, bPAG and cPAG, was measured in milk and in serum the 

concentration of bPAG was determined. Blood samples were taken 

from the jugular vein (v. jugularis) and 50 ml milk samples were 

taken and stored at -20 °C. 

Results: the profile of bPAG and cPAG concentration in milk 

increased after 28 day of pregnancy and showed a rapid increase close 

to parturition. The same results we observed in the blood concentration 

of bPAG. The drop of bPAG concentration was faster in the milk than 

in the blood. We found a good correlation between the results from the 

blood, milk, rectal palpation and USG pregnancy detection. The 

investigations showed that the concentration bPAG and cPAG in milk 

were not higher than 10 % of the bPAG concentration in the blood. 

Analysis of variance presented the correlation between pregnancy and 

cPAG concentration in milk (P < 0.1), and its higher than with bPAG in 

the blood plasma, what suggests lower correlation. 

Conclusion: our data showed that the RIA method was very precise 

for measuring PAG concentrations in the maternal blood and milk of 

the investigated cows. The results showed the possibility of the use 

of cPAG in milk and bPAG and cPAG in blood as pregnancy test to 

replies the traditional rectal investigation or USG method. 

Key words: PAG, ultrasonography, cows 

855 Efficiency of Different Cloprostenol Doses in the Postpartum 

Period of Zebu (Bos Taurus Indicus) Beef Cows 

C. Fernandes 1,2 , B. Alves 2 , E. Oliveira 2 , J. Viana 3 , A. Figueiredo 1,2 , 

M. Gioso 1 , E. Oba 4 

1 

University of Alfenas, Veterinary Medicine, Alfenas-MG Brazil, 

Brazil 

2 

Biotran, Veterinary Medicine, Alfenas-MG Brazil, Brazil 

3 

Embrapa Gado de Leite, Animal Reproduction, Juiz de Fora - MG - 


4 

UNESP- School of Veterinary Medicine and Animal Sciences, 

Animal Reproduction, Botucatu, Brazil 

Reestablishment of reproductive activity after parturition depends on 

two physiological processes, uterine involution and reestablishment 

of the ovarian luteal cyclic activity. Impaired or delayed uterine 

involution can also affect ovarian activity. Prostaglandins have an 

important function in this process. The prostaglandin F 2a (PGF 2a), 

normally produced during uterine involution, stimulates uterine 

muscular layer activity. The use of PGF 2a synthetic analogous in 

bovine postpartum, however, has been limited. Many studies 

demonstrate a beneficial effect of synthetic prostaglandins in the 

postpartum period of cows, but it is still not clear whether their 

isomers would have a similar effect. The kinetic of cloprostenol 

isomers can be different in the receptors located in the uterus or in 

the corpus luteum. The aim of this study was to compare the effect of 

two different doses of a racemic cloprostenol mixture (D+L- 

Cloprostenol) given in the postpartum period, on reproductive 

performance of Nelore (Bos taurus indicus) cows. The study was 

performed in a farm in Minas Gerais State, Brazil. Beef cows with 

normal parturition were evaluated and randomly distributed into 3 

groups: G1 (n=144), Control Group; G2 (n=145), 0.530mg of D+L- 

Cloprostenol, given IM at 3 to 5 days after parturition, and G3 

(n=145), 1.06 mg of D-Cloprostenol, in the same schedule as in 

group 2. Parameters evaluated were: services per conception (x 2 ), 

days from parturition to first estrus, and days open (Tuckey’s test).

There was no difference in body condition score among groups at 

parturition or during the postpartum period (P>0.05). There was also 

no difference in the number of services per conception. The average 

number of days from parturition to first estrus were 73.38±15.56 a ; 

64.42±17.05 b and 65.34±18.31 b , and the average number of days 

open were 97.34±26.54 a , 86.38±28.81 b and 85.23±30.12 b for groups 

1, 2 and 3, respectively (P

groups according to their diameter : 1 (2-5 mm.), 2 (>5-10 mm.), 3 ( 

>10-15 mm.) and 4 (>15-25 mm.), follicular fluids were aspirated by 

using disposable syringes , and mixed the fluids of the follicles 

which have the same size that collected in the same day, then kept in 

tubes under -20 °C until chemical assay. The results of study have 

shown that the concentration of Sodium decreased significantly in 

small follicles and it increased with the increased size of the follicle 

, but there was no significant difference between the last three sizes 

and the highest concentration of Sodium were noticed in Autumn, 

while the concentration of the Potassium, Calcium and Phosphorus 

increased significantly in the fluids of the small size follicles and 

they decreased in the larger follicles , also the highest concentration 

of Potassium, Calcium were noticed in autumn, and spring for 

Phosphorus. 

860 Lead Toxicity Leads Sperm Dysfunction in Holstein Bulls 

M. Arabi 

Shahrekord University, Biology, Shahrekord, Iran 

Lead (Pb) is a xenobiotic metal with no known essential function in 

cellular growth, proliferation, or signaling. Decades of research 

characterizing the toxicology of Pb have shown it to be a potent 

toxicant in the male reproductive system. Lead exposure occurs 

mainly through the respiratory and gastrointestinal tracts. 

Approximately 30-40 percent of inhaled lead is absorbed into the 

bloodstream. The objective of the present investigation was to study 

the effect of Pb in the concentration range 10-100 µM, on the 

membrane and DNA integrity, viability, and acrosome reaction in 

bull spermatozoa, in vitro. The semen dosages were obtained from 

healthy Holstein bulls. The lipid peroxidation/LPO assay assessed 

the membrane integrity as malondialdehyde/MDA levels. The 

viability test used for showing the percentage of live and dead bull 

sperm cells. The gelatin digestion test (GDT) was also used for 

detection of halos. The single cell gel electrophoresis technique 

detected the percentage of sperm DNA breaks/damages as 

comets.We showed a significant increase in LPO, concentrationdependent, 

indicating deleterious effect of Pb on the sperm 

membrane integrity. There was a negative correlation between LPO 

rate and percentage of viable spermatozoa (r=-0.92, p

H-hCG and H-saline subgroups (41.46% versus 24.81%, p

improve embryonic development are manipulating dairy cattle 

genetics to develop a cow that is more resistant to heat stress. 

867 Effect of Inserting CIDR Device from Day 28 to 35 following 

AI on Pregnancy Rate in Dairy Cows 

N. Shams Esfandabadi 1 , A. Shirazi 1 , P. Mirshokrai 1 , 

H. Ghasemzadeh Nava 2 

1 

Veterinary Faculty of Shahrekord University, Clinical Sciences, 

Shahrekord, Iran 

2 



Objective of the study: Reproductive inefficiency in lactating dairy 

cow is a source of frustration to dairy producer and their consultants 

and it reduces dairy farm profitability. Early embryonic mortality in 

cows returning to estrous before 24 days after insemination is 

approximately 20.5-43.6% and late embryonic mortality in cows 

returning to estrous after 24 days is approximately 8.0-17.5%. It was 

hypothesized that suboptimal progesterone concentrations during the 

late embryo and early fetal period may act to compromise conceptus 

development in dairy cattle. This study was conducted to evaluate the 

effect of inserting CIDR device from day 28 to 35 following AI on 

conception rate in dairy cattle. 

Material and method: Cows entered the study matched by parity, 

milk yield, days of lactation, and number of AI. All cows were 

inseminated at estrous by an on farm technician. Cows that were 

detected in estrous during the first period (days 18-24) were 

reinseminated and recorded as non pregnant to the previous AI. 

Cows which did not show estrous (n=74) were alternately assigned 

into two groups: treated group (n=37) received CIDR (containing 

1.38 gr progesterone, Pharmacia and Upjohn, Mt Wellington, 

Auckland, NZ) on day 28 for 7 days, and control group (n=37) 

received no treatment. They were observed for estrous in the second 

period (days 36-42). Pregnancy was diagnosed by rectal palpation on 

day 45- 50 after AI. The differences in pregnancy proportion 

between treated and control cows were analyzed by using Chi- 

Square and Fisher tests. 

Results: Pregnancy proportion in treated and control groups were 

56.7% and 51.3% respectively. The results demonstrated that there was 

no significant difference between the two groups (P>0.05). The effect 

of this treatment on pregnancy outcome was statistically indifferent 

among lactation, milk yield, days of lactation and number of AI. 

Conclusion: The treatment of cows with CIDR from 28-35 days after 

insemination did not improve conception rates. Further study is needed 

to determine the role of post insemination progesterone supplementation 

on conception rate in cows. 

Key words: dairy cow, CIDR, AI, pregnancy rate 

868 The Effects of GnRH or CIDR Administration on Pregnancy 

Rates in Dairy Cows during Summer Heat Stress 

GH. Moradli 1 , A. Moghaddam 2 , M. Hashemi 3 , M. Tymori 3 , 

M. Kazemi 3 

1 

Islamic Azad University, Saveh-Branch, Department of Clinical 

Science, Saveh, Iran 

2 

Razi University/Faculty of Veterinary Medicine, Department of 

Veterinary Medicine, Kermanshah, Iran 

3 

Islamic Azad University, Saveh-Branch, Department of Animal 

Science, Saveh, Iran 

Dairy cattle experience reproduction difficulties when exposed to 

conditions causing excessive heat stress. Thermal stress in cattle 

results in major decreases in dairy production and reproduction each 

summer. These decreases have been documented in many studies 

and reviews. Common methods to reduce the negative impact of heat 

stress on reproduction are to provide cooling in the form of shades, 

soakers, fans, or evaporative coolers as well as using embryo 

transfer, induction of accessory corpora lutea and crossbreeding. The 

objective of the present study was to evaluate protocols for 

improving pregnancy rates in dairy cows during summer heat 

stress.Estrus cycles of Holstein cows were synchronized using two 

i.m. injection of prostaglandin (PGF2a) 12 d apart, and cows were 

artificially inseminated (AI) 12 h after estrus detection. After AI 

(Day 0), the cows were randomly divided into groups 1 (Control; 

n=30), 2 (GnRH; n=30), 3 (CIDR; n=30) and 4 (CIDR / GnRH; 

n=30). Cows in group 1 did not receive any treatment. On Day 8 


postinsemination, cows in group 2 received GnRH agonist, while 

cows in group 3 treated with CIDR. Group 4 cows were given CIDR 

and GnRH at the same day. CIDR was removed at day 21 after AI. 

Blood samples were taken on days 0, 8, 16 and 21 days after AI. 

Cows were palpated per rectum at 45 days after AI to determine 

pregnancies. Serum progesterone (P4) concentration was higher 

(P

causes of congenital decapitation of spermatozoa, sperm motility in 

the moufflon was reduced, but the flagella were observed to move 

without the heads. Decapitation affected 17 to 57 % of the 

spermatozoa and most frequently it was accompanied by a 

protoplasmic drop, located proximally. Further developmental 

changes, affecting the flagella of the sperms, were observed in the 

moufflon. The mentioned morfological changes led to infertility in 

the moufflon and to a substantial reduction of fertility. After natural 

mating of the moufflon with a superovulated moufflon sheep, the ten 

eggs obtained included four blastocysts, two were degenerated, and 

four were unfertilized. 

(Funded by VEGA 1/3484/06 and AV 4/0009/07) 

Key words: moufflons, spermatozoa, fertility 

871 Preliminary Results on the Prevalence of Cytological 

Endometritis in Dairy Herds in North-east Poland 

W. Baranski, M. Podhalicz, T. Janowski, S. Zdunczyk, 

G. Lukaszewicz 

University Of Warmia And Mazury, Animal Reproduction, Olsztyn, 


It has been suggested that the clinical endometritis is an important 

cause of decreased fertility in Holstein cows. This disorder is 

diagnosed by cytological control of uterine smear, however 

information about its ethiopathogenesis and prevalence is still very 

limited. The aim of our study was to assess the prevalence of 

cytological endometritis (CE). 39 cows from 2 farms (A,B) without 

clinical endometritis were included in this trial. Cows were 

examined rectally and ultrasonographically for diameters of both 

uterine horns and cervix as well as presence of fluid in the uterus in 

fourth and sixth week post partum (pp). Samples for cytological and 

bacteriological examinations were collected from the lumen of a 

bigger horn by cytobrush placed in a stainless steel tube. The smears 

have been stained with Papanicolau staining and evaluated under 

light microscope. CE was diagnosed by increased number of 

polimorphonuclear neutrophils (PMN) in the smear over 18% of 

cells in fourth week pp, whereas in the sixth week the threshold was 

10%. According to the criteria during the first examination, CE was 

found in 39,3% and 81,8% of cows from farms A and B, 

respectively. Two week later significant decrease in the number of 

CE cows was observed, because only 21,4% of the animals from 

farm A and 36,4% from farm B were diagnosed as positive. Bacterial 

infections were not fully correlated to CE because bacteria were 

found in 35,7% of cases at farm A and 27,3% at farm B during the 

first examination and all samples were negative in the second 

examination. By rectal and ultrasonographical examination neither 

ovarian nor uterine abnormalities were observed during that period. 

These preliminary results indicate, that cytological endometritis is a 

common problem in dairy cows at the end of uterine involution. The 

relationship between CE and fertility parameters will also be 

investigated in a further study. 

Key words: cows, cytological endometritis 

872 Urea in Milk and Nutritional and Reproductive State of Dairy 

Cows 

G. Santos 1 , P. Grande 1 , H. Ribeiro 2 , J. Damasceno 2 , C. Alcalde 1 , 

O. Barbosa 1 , J. Horst 3 , F. Santos 1 

1 Universidade Estadual de Maringá, Zootecnia, Maringá, PR, Brazil 

2 Clínica Pioneiros, Ruminantes, Carambeí, PR, Brazil 

3 Associaçio Paranaense dos Criadores de Bovinos da Raça 

Holandesa (APCBRH), Controle Leiteiro, Curitiba, Brazil 

This study aimed at assessing the effects of Farms, parturition order 

and month of parturition on milk production, % fat and protein, 

somatic cell count and milk urea nitrogen (MUN). The effects of 

MUN on the gestation period and the anestrous in the beginning of 

lactation, rate of inseminated cows, parturition; insemination 

interval and the number of inseminations on four dairy Farms were 

also studied. This work was developed on four dairy Farms in Brazil. 

The Farms, nominated A, B, C and D, were under the dairy control. 

The data obtained from each Farm enabled the grouping of cows 

according to parity number and month of parturition to assess milk 

production and quality and concentration of milk urea (MUN). Three 

groups were formed to look at the effect of parity number: Group 1: 

animals of 1 st parity; Group 2: animals of 2 nd parity; Group 3: 

animals of 3 rd or greater parity. There were 35 heifers in group one, 

30 cows in group two, and 55 cows in group three. The data were 

collected from July to December 2002. Animals were blocked in 

four different classes to look at the effect of MUN concentration: 1. 

animals with levels lower than 10.0 mg/dL; 2. animals with levels 

between 10.1 and 13.0 mg/dL; 3. animals with levels between 13.1 

and 15.0 g/dL; and 4. animals with levels higher than 15.0 mg/dL. 

The obtained data were analyzed using the software SAS and the 

means were compared through the Tukey Test at 5%. With the 

exception of the comparison of the MUN concentrations per band 

over the % of artificially inseminated cows (AIR), % of cows in 

estrus, gestation rate and parturition; AI interval, which underwent a 

Kruskal-Wallis non-parametric test. Also, the effects of MUN on 

gestation rate (GR) AIR in early lactation were studied. The 

parameters of production and milk quality (34,86 kg milk/d; 3,0 and 

2.83%, fat and protein) were similar among farms and were not 

influenced by the month of parturition. Farm D presented the highest 

MUN concentration (14.60±0.46 mg/dL). Concentration of MUN, 

percentages of fat and protein and SCC were not influenced by PO. 

Cows with a value of MUN between 10.1 and 13.0 mg/dL had the 

highest AIR and GR between days 55 and 70 postpartum and after 70 

days in milk. These data suggest that MUN concentration is a useful 

parameter to predict the nutritional and reproductive stages of dairy 

cows. 

The project was supported by the CNPq, Brasilia, DF and Fundaço 

Araucária - Curitiba, Paraná 

Key words: dairy cattle, gestation, insemination, MUN 

873 Hysteroscopic Picture of Postparturient Metritis in Cows 

R. Mordak, K. Kubiak, M. Jankowski, J. Nicpon 

Wroclaw University of Environmental and Life Sciences, Animal 


Hysteroscopy and endometrial ablation techniques are quite popular 

in human medicine. In bovine practice hysteroscopy in cows is 

occasionally used. Commonly used methods for detecting uterine 

pathological features in cows are gynecological examinations via 

vaginal or rectal palpation. These methods are usually sufficient for 

an adequate diagnosis and therapy in cows. The aim of the paper was 

the use of a flexible fiberoscope for hysteroscopic examinations in 

cows, manifesting clinical signs of postpartum metritis after placenta 

retention in their early period of uterine involution. Seven Polish 

Holstein-Friesian (black and white) cows after pathological delivery 

were examined using hysteroscopy. Retention of fetal membranes 

was observed in those cows. Retention of fetal membranes were 

manually resolved with a consecutive antibiotic intrauterine 

protection. A week after manual removal of fetal membranes (9- 10 

days post calving) the uterus was clinically examined, including 

hysteroscopy. A fiberoscope, type Olimpus GIF XQ-20 with a 

working length of 1000 mm and outside diameter of 9,8 mm, was 

used. This paper shows eight, selected, the most characteristic 

hysteroscopic pictures of the uterus of cows with metritis, taken 

during clinical examinations (after fetal membrane retention). The 

pictures were recorded respectively through vulvar cleft, than 

cervical canal, uterine body, uterine septum to the horns. They 

involves pathological lesions in endometrium as well as a view of 

pathological discharges. 

874 Pregnancy Rate in Anestrous Bos Taurus/Bos Indicus 

Crossbred Cows Given a CIDR Insert and Estradiol, with or 

without an Injection of Progesterone 

C. Lamothe 1 , F. Montiel 1 , R. Cuicas 2 

1 Universidad Veracruzana, Facultad de Medicina Veterinaria y 

Zootecnia, Reproducción, Veracruz, Mexico 

2 Universidad Autónoma de Yucatán, Facultad de Medicina 

Veterinaria y Zootecnia, Reproducción, Mérida, Mexico 

Objectives of the study: To evaluate the pregnancy rate in postpartum 

anestrous lactating Bos taurus/Bos indicus crossbred cows after 

treatment with 1.9 g of natural progesterone contained in an 

intravaginal insert (CIDR, controlled internal drug-releasing device), 

estradiol and synthetic progesterone (P 4). 

Materials and Methods: On days -9, -6, -3 and 0 (day 0=start of 

treatment), anestrus status was confirmed by transrectal ultrasonography 

and determination of serum P4 concentrations (

in each sample). On day 0, 104 cows were assigned to the following 

treatments: 1) CIDR+ECP+P4 (n=24): one CIDR insert + i.m. 2 mg 

of estradiol cypionate (ECP) and 100 mg of P 4; 2) CIDR+ECP 

(n=22): a CIDR + 2 mg of ECP; 3) CIDR+EB+P4 (n=20): a CIDR + 

i.m. 10 mg of estradiol benzoate (EB) and 100 mg of P 4; 4) 

CIDR+EB (n=21): a CIDR + 10 mg of EB; 5) Control (n=17): i.m. 5 

ml of saline solution as a placebo. The CIDR inserts were removed 

on day 9. The CIDR-treated cows received timed artificial 

insemination (TAI) 48 to 56 h after its removal, and the control cows 

were AI 12 h after detected estrus. Detection of estrus was performed 

twice daily from 24 h after CIDR removal until three days later. 

Results: Pregnancy rate was 58.3% (14/24) for CIDR+ECP+P4, 

63.6% (14/22) for CIDR+ECP, 45% (9/20) for CIDR+EB+P4, 

47.6% (10/21) for CIDR+EB (P>0.05) and 17.6% (3/17) for the 

control group (P

spermatozoon, average value). The same tendency was registered 

both in the case of thermo resistance test during a period of three 

hours of incubation at 37 °C and in the case of acrosomal integrity 

distinguished through the FITC-PNA/PI-staining and examined at 

flow cytometry. Acrosomal integrity was better (p0.05 ). Altogether, from 50 cases of pregnancy, 28 cases (56%) 

were in the right horn and 22 cases (44%) were in the left horn. It is 

concluded that insemination of cattle with left ovarian ovulation may 

increase the rate of female fetuses at pregnancy and parturition. 

However, this effect has not been observed in ewes. 

Key words: sex ratio, sexing, left and right uterine horn pregnancy 

879 Detection of Leptospira spp. in Semen and Vaginal Fluids of 

Goats and Sheep by Polymerase Chain Reaction 

W. Lilenbaum 1 , R. Varges 1 , F. Brandao 1 , A. Cortez 2 , S. Souza 2 , P. 

Brandao 2 , L. Richtzenhain 2 , S. Vasconcellos 2 

1 

Universidade Federal Fluminense, Microbiology, Niterói - RJ - 


1 

Universidade de Sao Paulo, Preventive Medicine and Public Health, 


The purpose of the present study was to evaluate the use of PCR for 

the detection of Leptospira spp. in semen and vaginal fluids of goats 

and sheep. Thirteen goat herds and seven sheep flocks in the state of 

Rio de Janeiro, Brazil were screened for leptospirosis, using 

serologic approaches. In this first step, approximately 20% of the 

adult animals in each herd/flock were randomly selected; overall, 

248 caprine and 292 ovine serum samples were tested by a 

microscopic agglutination test - MAT. From those, three herds and 

three flocks with great proportion of seroreactive animals (>30% in 

each herd/flock) were identified, and 19 goats (16 females and three 

bucks) and 40 sheep (26 ewes and 14 rams) that were seropositive 

(specific anti-Leptospira titres >400, based on MAT), were selected 

for more detailed studies. From those animals, samples of vaginal 

fluids or semen were collected for bacteriological and molecular 

assays. Diluted semen and vaginal fluid samples were centrifuged 

(3000 x g for 10 min) and the supernatant used for both molecular 

and bacteriological assays. Blood samples were also centrifuged 

(1000 x g for 10 min) and examined for Leptospira antibodies by 

MAT. Bacterial DNA was extracted by a phenol and guanidine thiocyanate 

method. The PCR assay for the detection of Leptospira spp. 

is genus-specific and based on protocol that employs the primers 

Lep1 (5’-GGCGGCGCGTCTTAAACATG-3’) and Lep2 (3’- 

TTAGAACGAAGTTACCCCCCTT-5’). For both species of 

animals, the most common reactions were to serovars Hardjo, 

Shermani, and Grippotyphosa. Although leptospires were detected 

by darkfield microscopy in three vaginal fluid samples (from two 

goats and one ewe), pure isolates were not obtained by 

bacteriological culture of semen or vaginal fluids. However, seven 

vaginal fluid samples (four goats and three ewes) and six semen 

samples (all from rams) were positive on PCR. Based on these 

findings, in addition to analogous findings in cattle, we inferred that 

there is potential for venereal transmission of leptospirosis in small 

ruminants. 

Key words: leptospirosis, PCR, reproduction, sheep, goats 

880 Use of 3D-Cell Culture of Bovine MEC in Study of the Role of 

Hormones and Growth Factors in the Formation of 

Differentiated Acinar Structures 

M. Gajewska, M. Kozlowski, P. Jasinski, K. Hajduk, T. Motyl 


Department of Physiological Sciences, Warsaw, Poland 

Mammary gland epithelium is comprised of individual acinar units, 

which are notable for hollow lumen, surrounded by polarized 

epithelial cells. The development and maintenance of this polarized 

structure is critical for the form and function of epithelial 

cells.Mammary epithelial cells supported on a laminin-rich 

extracellular matrix (ECM) form three-dimensional (3D) acinar 

structures that mature to form polarized and functional monolayers 

surrounding a lumen and having the ability to produce milk proteins. 

We have used the 3D laminin-rich ECM cell cultures to study the 

regulation of bovine mammary epithelial cells (MEC) differentiation 

in vitro, on the model of BME-UV1 cell line. The role of hormones 

(prolactin - PRL, growth hormone GH, and progesterone) and 

growth factors (EGF, IGF-I and TGF-1) was examined in the context 

of MEC polarization and functional differentiation. 

Immunofluorescence, confocal microscopy and Western-blotting 

techniques were used in order to determine the expression of chosen 

polarization markers: E-cadherin and ZO-1, and a marker of 

functional differentiation: milk protein L-casein. 

Western-blot analysis showed an increased expression of L-casein in 

bovine MECs cultured in ECM environment in comparison to a 

classical monolayer cell culture. The 3-D cell culture conditions also 

exhibited an increased expression of E-cadherin and ZO-1 proteins. 

Confocal images of immunocytochemical staining revealed that 

proper polarization of cells forming 3D-acinar structures was 

obtained when hormones: PRL, or GH or progesterone were added to 

the medium. However administration of growth factors: IGF, EGF or 

TGF-1 caused a disturbed pattern of E-cadherin and ZO-1 staining, 

which may suggest a failure in proper polarization of acinar 

structures cultured on ECM. 

The results of our study show that all used hormones exhibit a positive 

role in the formation of fully polarized 3D-acinar structures, with a 

hollow lumen and the ability to produce L-casein. The addition of EGF 

or IGF-I to the cell culture led to a partial failure in cell differentiation, 

causing a prolonged cell proliferation and not full lumen clearance, 

while TGF-1 treated cells showed morphology of small disorganized 

spheres. To summarize, our study has shown that the process of acini 

formation by bovine MEC requires the involvement of studied 

hormones, especially PRL and GH. 

Key words: 3D-MEC culture, growth factors, prolactin, GH, 

differentiation 

881 Regulation of Autophagy in Bovine Mammary Epithelial Cells 

A. Sobolewska 1 , M. Gajewska 1 , J. Zarzynska 2 , B. Gajkowska 3 , 

T. Motyl 1 

Reproduction and Biotechnology 197

1 



2 


Department of Food Hygiene and Public Health, Warsaw, Poland 

3 



Mammary gland growth and involution is based on a dynamic 

equilibrium between proliferation and apoptosis of mammary 

epithelial cells (MEC). The main type of cell death responsible for 

bovine mammary gland involution is apoptosis, but also a lot of cells 

exhibit morphological features of autophagy. Autophagy is a highly 

conserved catabolic process responsible for degradation and 

recycling of cellular constituents (long-lived proteins and 

organelles) by lysosomes. The highest intensity of autophagy in 

MEC was observed during dry period, when the gland undergoes 

intensive remodeling and the deprivation of bioactive compounds 

and nutrients occur. Present study has shown the role of mTOR in 

the regulation of autophagy by examined growth factors (IGF-I, 

EGF) and hormones (progesterone and 17‚-estradiol) in bovine 

MEC. In our study we have used a model of bovine mammary gland 

involution in vitro, dependent on FBS deficiency in the culture of 

bovine BME-UV1 cells. Immunofluorescene, electron microscopy, 

laser scanning cytometry, LysoTracker-Green related fluorescence 

and Western-blot were used as methods for analyzing lysosomes 

activation, expression of MAP1LC3, phospho-mTOR and active 

caspase-3 as biochemical markers of autophagy and apoptosis, 

respectively. FBS deficiency induced both autophagy and apoptosis 

with the highest intensity of both processes at the 48 h of MEC 

exposure to the deficiency medium (0.5% FBS). EGF completely 

abolished autophagy induced by FBS deficiency. A similar but less 

pronounced effect of autophagy suppression was observed in case of 

IGF-I. We have shown that inhibition of mTOR by rapamycin (a 

specific inhibitor of mTOR) abrogated suppressive effects of IGF-I 

and EGF on FBS deficiency-induced autophagy, which suggests that 

mTOR is an important link in IGF-I ad EGF-dependent antiautophagic 

pathways. Conversely to IGF-I and EGF, 17‚-estradiol 

and progesterone exerted stimulatory effects on autophagy in bovine 

MEC. At the same time a suppressive effect of both steroids on 

mTOR activation/phosphorylation was observed. In conclusion, 

autophagy in bovine MEC undergoes complex regulation, where the 

principal function belongs to survival pathways dependent on IGF-I 

and EGF, which are involved in the suppression of autophagy, while 

pregnancy steroids show the role of inducers of this process. 

Key words: autophagy, mammary gland, growth factors, sex steroids, 

mTOR 

882 Effects of Serum Calcium (Ca) and Inorganic Phosphorus (Pi) 

Levels during Transition Period on Reproductive 

Performance of Dairy Cows 

F. Di Ciommo 1 , E. Parmigiani 1 , E. Bigliardi 1 , G. Morini 1 , F. Righi 2 , 

F. Di Ianni 1 , L. Ferrari 1 , C. Bresciani 1 , I. Vecchi 1 , A. Di Ciommo 3 

1 

College of Veterinary Science, Animal Health Department, Parma, 

Italy 

2 

College of Veterinary Science, Nutrition, Parma, Italy 

3 

Practitioner, Parma, Italy 

This study was designed to evaluate the influence of Ca and Pi serum 

levels during transition period on reproductive efficiency of dairy 

cows fed two close-up rations Dietary Cation-Anion Difference 

(DCAD). The study was conducted on 43 multiparus Holstein cows 

aging 48 ± 16 months. Data were collected from day -20 to day +20 

from parturition during the winter. Cows were distributed in two 

groups: Group A (n = 20) fed a basic close- up diet added with 

1Kg/d/cow clorine and sulphates anionic salts (DCAD = 5 Meq); 

Group B (n=23) fed a basic close-up diet (DCAD =30 Meq). Blood 

samples were taken from jugular vein at 20, 10 and 2 days before the 

expectated calving day, at calving and 2, 10 and 20 days after 

parturition to evaluate serum Ca and Pi levels. On days 20 and 2 

prior calving also urine samples were collected to estimate pH trend 

in both groups. In group A the urine pH dropped significantly from 

20 to 2 days before calving (from 8.15 ± 0.36 to 6.9 ± 0.51; p < 0.05). 

In group B there were no changes in urinary pH. Ca level during 

transition period was similar in the two groups and reached the 

lowest level at the day of calving day (group A: 6.02 ± 0.98 mg/dl; 

group B 6.1 ± 1.19 mg/dl), while Pi level was significantly higher, 


reaching the highest level at day 2 after calving (group A 6.02 ± 

1.02; group B 5.13 ± 1.1 mg/dl; p < 0,05). Calving-first service 

interval was shorter in Group A than in Group B (96.5 days ± 16.95 

vs 115.17 ± 55.31 respectively). Cows in group A had significantly 

fewer days open than cows in group B (109.2 days ± 30.35 vs 140.09 

± 39.29 respectively; p < 0,05). Services for Conception were lower 

in group A than B (1.5 vs 2.1, respectively). First service-conception 

rate was higher in group A than in group B (50% vs 39.13%, 

respectively). The incidence of retained placenta was lower in group 

A than in group B (15% vs 31.82%, respectively). In this study cows 

treated with anionic showed better reproductive performance, even if 

they had similar total calcium levels as untreated animals. These 

results suggest that the use of anionic salts during close-up period 

can improve reproductive performance of dairy cattle and, according 

to other authors, the level of calcium in the ration should be 

increased proportionally to DCAD reduction. 

883 Increased Frequency of Stillbirth in a Holstein-Friesian Herd: 

Birth Process 

M. Hoedemaker, Y. Gundelach, K. Essmeyer 

University of Veterinary Medicine Hannover, Clinic for Cattle, 

Hannover, Germany 

It was the aim of this study to identify possible causes of stillbirth on 

a large dairy farm in Germany. Four hundred and sixty-three cows 

with 483 calves born were observed continuously starting from the 

time when the animals were moved into the “reproduction barn” 

until completion of parturition. Upon first signs of the forthcoming 

labour, every action of the dam and the barn staff were documented. 

The interval from rupture of the allantois/amnion sac until passage of 

the fetal forehead through the vulva (second stage of labour (SSL)) 

and the duration of final expulsion of the fetus (third stage of labour) 

were recorded. The birth process was classified in five categories 

(1= spontaneously, 2= light extraction, 3= medium extraction, 4= 

severe extraction, 5= caesarean section) and the positions of the fetus 

were documented. Overall, 9.3% of all births (n=43) were classified 

as stillbirth. Frequency of stillborn calves was lower in birth 

category 1, 2, and 3 than in category 4 and 5 (2.3%, 4.8% and 7.4% 

vs. 28.4% and 41.2%, respectively, P

4=severe extraction, 5= caesarean section) and the positions of the 

fetus were documented. After birth, the following data were 

collected from the calves: vitality, sex, weight and various 

measurements of the fetlock joint and head. Calves born dead and 

those that died within the first 24 h after birth, were classified as 

“stillbirth” and were subjected to a pathomorphological, histological 

and microbiological examination. 9.3 % of all births (n=43) were 

classified as stillbirth. Twin births were associated with a higher 

frequency of stillbirth than the birth of single calves (17.7 % vs. 9.0 

%, P

and durable spermatozoa. Within the same season (BS or Non BS) 

significant differences are recorded in ejaculates of different motility 

status (p0.001). This fact clearly suggests that the stability of 

their cell membrane is rather a seasonal attribute than is related to 

motility. 

Key words: ram semen, semen kinematics, seasonality 

888 Expression of Innate Immune Genes in Bovine Peripheral 

Blood Leucocytes (PBL): Relevance to Endometritis in postpartum 

Dairy Cows 

A. Chapwanya 1 , K. Meade 1 , M. Doherty 2 , J. Callanan 2 , J. Mee 3 , 

C. O’Farrelly 1 

1 

Trinity College Dublin, School of Biochemistry and Immunology, 

Dublin, Ireland 

2 



3 

Teagasc, Dairy Production, Fermoy, Ireland 

Infectious bovine endometritis is the most common reproductive 

disease of post-partum cows. In the immediate post-calving period, 

all cows experience an influx of bacteria. Most cows will eliminate 

all bacteria within 6 weeks of calving, but about 20% fail to do so 

resulting in persistence and development of uterine disease. 

However, little is known regarding the role of the uterine innate 

immune response in dairy cows post-partum. We have recently 

described 11 novel antimicrobial peptide (AMPs) genes in the 

bovine genome. This study was designed to examine the expression 

of these and other innate immune genes in PBL from post-partum 

dairy cows. PBL were obtained from 12 dairy cows (healthy and 

endometritic), RNA extracted and cDNA synthesised. Primers were 

designed for selected genes involved in pathogen detection and 

innate immune response to bacterial infections, including Toll-Like 

Receptors (TLRs), cytokines and AMPs. The expression of these 

genes was analysed by quantitative real-time PCR (qRT-PCR). 

Preliminary results from 2 dairy cows show that known AMPs - LAP 

and TAP are up-regulated in PBL from cows with endometritis. 

BNBD4 expression remains unchanged in infection while the 

expression of BNBD5 is reduced. Altered expression of innate 

immune genes in bovine PBL may be a useful prognostic indicator of 

uterine infection. 

Key words: endometritis, AMPs, innate immunity, PBL 

889 Susceptibility of ST, MDBK and BHK Cell Lines to Infection 

with BoHV-1, bPIV3, BRSV and BVDV-1 

R. Jordao 1 , C. Ribeiro 2 , E. De Stefano 2 , C. Del Fava 2 , C. Camargo 2 , 

E. Pituco 2 

1 

Instituto Biológico, Laboratório de Produçao de Imunobiológicos, 


2 

Instituto Biológico, Centro de Pesquisa e Desenvolvimento de 

Sanidade Animal, Sao Paulo, Brazil 

Many viruses are potential contaminants of biological products 

produced in bovine, porcine or hamster cells or manufactured via 

processes using animal-derived raw materials such as bovine serum. 

The susceptibility of cell lines was tested by inoculation with four 

bovine viruses. Cell lines Swine Testicle (ST), Madin Darby Bovine 

Kidney (MDBK) and Baby Hamster Kidney (BHK), were 

experimentally infected with BoHV-1, bPIV-3, BRSV and BVDV-1 

to determine if they were permissive for virus replication. To 

determine whether cells were susceptible to viral infection, each cell 

line was infected with one strain and incubated in 5% CO 2 at 37 °C, 

until cytopathic effect (CPE) was seen, or harvested after four days 

and were serially passaged until 32 days pos-infection (pi) with no 

evidence of CPE, after this period when submitted by PCR and 

ELISA for the presence of virus. ST cells showed evidence of CPE 


to BoHV-1 after 24 h (2nd passage - P2); to PI3 48h (1st passage - 

P1); BRSV 72h (P1) and BVDV-1 48h (P2) pi. MDBK cells showed 

evidence of CPE to BoHV-1 after 24 h (P1); to PI3 18h (P1); BRSV 

96h (P1) and BVDV-1 48h (P1) pi. BHK cells showed evidence of 

CPE to BoHV-1 after 48 h (P1); to PI3 48h (P1); BRSV 72h (P1) pi. 

In contrast, cells do not showed evidence of BVDV-1 infection after 

8th passage, ELISA assay and RT-PCR of experimentally infected 

BHK cells demonstrated that there was no evidence of infectious 

BVDV or contaminating viral genome. These results indicate that 

the cells susceptibility of a species are not completely predictable 

and that many cells derived from other species than cattle can be 

infected with bovine virus, except BHK that is refractory to BVDV 

infections. The presence of BVDV in cell cultures and in fetal bovine 

serum has long been recognized problem not only in laboratories but 

also among vaccine manufacturers. Consideration of the choice of 

cell lines used in the production of vaccines also should be based on 

the assessment of the risks of cross-contamination. In Brazil, a 

vaccine for foot and mouth disease has been prepared in the BHK 

cell line, and it is estimated that more than 395 million doses/year 

have been administered to cattle. Viral contaminations are a serious 

threat for animal cell cultures and may lead to false results in 

research, development, and virus screening, to viral contaminations 

in the biological products derived from the contaminated cultures. 

Key words: cell culture, contaminants, susceptibility, viral contaminant 

890 Effect of Mineral and Vitamin Treatment in Anovulatory 

Anestrous Dairy Cows 

M. Farhoodi, S. Mohamadi 

Islamic Azad University, Science & Research Branch, Faculty of 

Veterinary Medicine, Tehran, Iran 

Introduction: The anestrous syndrome is a common and economically 

important problem of world wide dairy farming. Anovulatory anestrous 

is a multifactorial problem. Treatment options for anestrous cows are 

different and include hormonal and management strategies and an 

efficient treatment method of anestrous in dairy cows are preferred. The 

objective of this study was to evaluate the effect of mineral and vitamin 

therapy in anovulatory anestrous dairy cows. 

Materials and Methods: The study was conducted in a large dairy 

herd of 750 Holstein cows. All cows were examined for their 

reproductive status 30 days postpartum. Cows, which had small 

ovaries without palpable structures in rectal examination ten days 

apart, were assigned as inactive ovaries anoestrous. These cows are 

mostly in their first and second lactation. These anoestrous cows 

were treated either with a single injection of GnRH (Buserelin) and 

PGF 2a (cloprostenol) ten days later (GnRH group, n=89 cows) or 

intramuscular phosphorus and Vitamins AD3E Therapy for three 

days then injection of GnRH in third day and PGF 2a ten days later 

(GnRH plus mineral-vitamin group n=58 cows). Reproduction data 

were collected and signs of estrous (cure symptom) after each 

treatment were recorded by visual observation (three times in day 

heat detection) and confirmed by rectal palpation, examing uterus 

and ovaries and checking mucous discharge. The curing rate was 

calculated in each group and analyzed with chi-square test. 

Results and Conclusions: In GnRH plus mineral-vitamin group 38 

out of 58 cows (65.5%) showed estrous and in GnRH group 51 out of 

89 cows (57.3%) showed estrous after treatment (p

from 3 dairy herds located in the vicinity of Tehran and with days in 

milk ranging from 34 to 44 days, were presynchronized by two 

injections of a PGF 2a analogue 14 days apart with the second 

injection given 12 days before initiating the synchronization 

protocols. Cows were assigned randomly to three treatment groups: 

Group1 (n=56), received conventional Ovsynch (GnRH, d 0; PGF 2a, 

d 7; GnRH 48 hours after PGF 2a and timed artificial insemination 

(TAI) 64 h after PGF 2a). Group 2 (n=53) received Cosynch 

64(GnRH, d 0; PGF 2a, d 7; GnRH and TAI 64 h after PGF 2a). 

Group3 (n=48) received Heatsynch (GnRH, d 0; PGF 2a, d 7; 

Estradiol benzoate 24 h after PGF 2a; TAI 64 h after PGF 2a). 

Pregnancy was diagnosed by palpation per rectum of uterine 

contents on d 40 to 42 after TAI. Results showed that pregnancy 

rates were greater for Cosynch 64 than for Heatsynch and Ovsynch 

protocols (32% vs. 21% and 26%, respectively). However overall 

pregnancy rates were not significantly different between the groups 

(p=0.469). Body condition scores (BCS) ranged from 1.0 to 5.0 on 

breeding day. Interaction between BCS and pregnancy rates revealed 

that pregnant cows had greater BSC than the open ones (2.3±0.05 vs. 

2.8±0.05 p

motility and CM integrity (p=0.869 and p=0.892 respectively, 

Friedman Test), in the three concentrations studied. Amicacine 

sulphate was confirmed as a good option for the control of semen 

microbiota mainly in IVF embryo production. 

This paper was supported by FAPESP: Fundaçio de Amparo a Pesquisa 

do Estado de Sao Paulo, Brazil 

Key words: amicacine sulphate, spermatozoa viability, membrane 

integrity 

895 Efficacy of a Treatment with hCG 4 Days after AI to Reduce 

Pregnancy Losses in Lactating Dairy Cows after 

Synchronized Ovulation 

C. Fischer-Tenhagen 1 , G. Thiele 1 , W. Heuwieser 2 , B. Tenhagen 3 

1 

Tierarztpraxis Thiele, Baruth, Germany 

2 

Freie Universität Berlin, Tierklinik für Fortpflanzung, Berlin, 

Germany 

3 

Bundesinstitut für Risikobewertung, Infection epidemiology and 

zoonoses, Berlin, Germany 

The objective of the study was to investigate, whether a treatment 

with hCG 4 d after AI could reduce pregnancy losses in high yielding 

dairy cows. Pregnancy rates (PR) and pregnancy losses were 

compared in four groups of cows. Cows of one large dairy herd 

presented to the veterinarian in a fixed reproductive management 

protocol were treated with an Ovsynch protocol if no corpus luteum 

(CL) could be palpated per rectum (Group OV). Cows with a CL 

received cloprostenol (0.15 mg) and buserelin (0.01mg) 2 d later and 

timed AI 16 to 20 hrs later (Group PG). Cows in both treatment 

protocols were then assigned to two groups by the final digit of their 

identification number (even vs. odd numbers) to receive 2,500 IU of 

hCG i.v. four days after AI (Groups OV-hCG, OV-Control, PG-hCG 

and PG-Control). Pregnancy diagnosis was carried out 27 d after AI 

via ultrasound and 39 d after AI by rectal palpation. Pregnancy 

losses were defined as cows being pregnant on d 27 but not pregnant 

on d 39 after AI. Pregnancy rate by d 27 did not differ between the 

four treatment groups (35.4, 35.0, 37.0 and 38.0 % for Groups OVhCG, 

OV-Control, PG-hCG and PG-Control, respectively). 

Pregnancy losses between d 27 and d 39 after AI were smaller in 

hCG treated animals in the summer months, but did not differ in 

autumn and spring. Pregnancy rate by d 39 after AI were higher in 

PG than in OV groups, but independent of hCG-treatment. In 

conclusion, treatment with hCG 4 d after AI did not significantly 

increase PR 39 d after AI. A positive effect of hCG on pregnancy 

losses during the summer months warrants further investigation. 

896 The Influence of Inmodulen on PMN and L activity and 

Antioxidative Potential in Blood of Cows 

H. Markiewicz, M. Gehrke, E. Malinowski, M. Kaczmarowski 

National Veterinary Research Institute, Department of Pathophysiology 

of Reproduction and Mammary Gland, Bydgoszcz, Poland 

The aim of the study was to establish of the best term of the second 

injection of Inmodulen and evaluate the influence of its on oxygenic 

metabolism of neutrophils and antioxidative potential in blood of 

cows in the puerperal period.Preliminary examinations were 

conducted on 20 healthy, non pregnant cows between 80 and 100 day 

of lactation. Ten cows received a single i.m. injection of 10 ml of the 

Inmodulen and the remaining 10 cows constituted the control group. 

The blood samples from experimental and control cows were taken 

from the tail vessels on day 0, 7, 14, 21. The level of both 

spontaneous and induced luminodependent chemiluminescence (CL) 

was determined. It was stated that single injection caused a 

significant increase of spontaneous and induced CL on the 7 th and 

14 th day (vs. the “0” day), which decreased on 21 st day after 

injection. The highest CL was detected on day 14, with exception of 

OZ (similar level was on day 7 and 14). On the basis of these results 

it has been suggested that the second application of the preparation 

should be made 14 days after the first application. The basic 

investigations were carried out on 35 cows: 18 experimental 

receiving Inmodulen and 17 controls. Inmodulen was injected on the 

day of parturition and on the 14th day thereafter. The blood was 

sampled once a week during the first 5 weeks of lactation. The level 

of CL, phenotype of subpopulation of T lymphocytes (CD2, CD4, 

CD8) with flow cytometry, the TAS of plasma as well as SOD, GPx, 

and CAT were examined. A significant influence was demonstrated 


of both the preparation and the time after parturition on the increase 

of the value of spontaneous and induced CL. The time of 

examination was important for CD2 lymphocytes, whereas the 

preparation and the time after parturition influenced CD4 

subpopulation. CD8 subpopulation did not change. The time of 

samples collecting had a significant influence only on the activity of 

TAS and SOD unlike GPX whose levels depended on the 

administration of Inmodulen.The level of TAS and SOD 

systematically increased from 1 to 5 week, unlike CAT whose level 

was the same all time. It was stated that Inmodulen i.m. injected on 

the day of parturition and 14 days thereafter, resulted in the 

significant increase of CL, which directly points on an increase of 

bactericidal activity of PMNs. The activation of these cells did not 

influence on the pro-antioxidative equilibrium of plasma and the 

activity of SOD and CAT, however, it increased of GPx activity. 

Key words: cows, PMN, chemiluminescence, antioxidative potential, 

Inmodulen 

897 Endometrial Changes in Postpartum Period in Cows 

I. Sematovica 1 , M. Pilmane 2 , A. Jemeljanovs 1 , D. Ikauniece 1 

1 

Research Institute of Biotechnology and Veterinary Medicine Sigra 

of the Latvia University of Agriculture, Sigulda, Latvia 

2 

Riga Stradins University, Institute of Anatomy and Antropology, 

Riga, Latvia 

The aim of the research was to reveal inflammatory factors, 

neuropeptide-containing innervation, distribution of growth 

stimulating, degradation factors, and apoptosis in the cows’ 

endometrium in postpartum period. Nine multiparious cows after 

normal calving were biopsied twice in the first and the fifth week 

after parturition. Matrix metalloproteinases (MMP2 un MMP9, 

1:100, R&D, England), tumor necrosis factor (TNF, 1:100, Abcam, 

England), interleukin 10 (IL10, 1:400, Abcam, England), vascular 

endothelial growth factor (VEGF, 1:50, DakoCytomation, 

Denmark), nerve growth factor receptors (NGFR p75, 1:150, 

DakoCytomation, Denmark), protein gene product (PGP 9.5, 1:1600, 

DakoCytomation, Denmark) were immunohistochemically detected, 

and the TUNEL method was used for detection of apoptosis (PCD). 

Haematoxylin and eosin staining was performed for each sample. 

Hyperemia, different level of infiltration of tissue with inflammatory 

cells, oedema, detachment of the lining epithelium, and erosive 

surface of mucous membrane in separate samples were observed in 

the first week after parturition. Four weeks postpartum, hyperplasia 

of basal cells, hyperemia, proliferation of endometrial glands, 

perivascular and periglandular infiltration with inflammatory cells, 

as well as lymphatic follicules were found. There were detected a 

significant increase of the number of inflammatory cells, TNF 

amount, and expression of VEGF, NGFR p75. A mild positive 

correlation was observed between the amount of inflammatory cells 

and TNF, PCD and TNF, TNF with NGFR p75, NGFR p75 and PGP 

9.5, VEGF and PCD, VEGF and NGFR p75. A strong positive 

correlation was seen between TNF and VEGF, as well as VEGF with 

NGFR p75. The increased expression of NGFR p75, VEGF and TNF 

seems to correlate with the ischaemia of tissue increased by 

inflammatory action in the cows endometrium from the first up to 

the fifth week postpartum. The latter also seems to stimulate the 

apoptosis and proliferation of nerve fibres in the cows endometrium. 

Key words: uterus, after parturition period, cows 

898 Efficacy of Three Antibiotic Protocols Added to Bull Semen 

Extender for the Inactivation of Leptospira Spp. Serovars 

Hardjo, Wolffi and Grippothyphosa 

M. Genovez 1 , T. Gotti 1 , V. Castro 1 , C. Horbach 2 , F. Viana 3 , 

S. Azevedo 4 

1 Instituto Biológico, Animal Health Center, Sâo Paulo, Brazil 

2 Universidade Estadual Paulista, Campus Botucatu, Animal 

Reproduction and Veterinary Radiology, Botucatu, Brazil 

3 Universidade Federal De Campina Grande, Department for 

Infectious diseases- Epidemiology, Patos, Brazil 

4 sersia Brasil, Animal Reproduction &Ai, Bragança Paulista, Brazil 

Two approaches can be applied to assure the safety of semen in 

AICenter: continuous surveillance of bulls before and after semen 

production and end product checking. The World Organization for 

Animal Health recommends adding at least one antibiotic treatment

to the extender: A-gentamicin (250 µg/mL), tylosin (50 µg/mL), 

lincomycin (150 µg/mL), spectinomycin (300 µg/mL); or Bpenicillin 

(500 UI), streptomycin (500 UI), lincomycin (150 µg/mL), 

spectinomycin (300 µg/mL). Some authors have shown that 

Leptospira spp. remain viable in semen after thawing, in spite of 

antibiotic treatment. Amicacine sulphate has been shown to be 

efficient against ubiquitous microbiota control when added to the 

semen extender, being easier to prepare and cheaper than treatments 

A and B. The present trial aimed at studying the efficacy of three 

antibiotic treatments A, B e C (amicacine sulphate -1,500 µg/mL) 

added to the semen extender for inactivation of Leptospira spp., in 

experimentally contaminated semen. Samples were collected from 8 

bulls (3-7 years old), all negative to leptospirosis, using an artificial 

vagina. Three hundred µL of semen were added to the extender 

treated with A, B or C antibiotic protocols and experimentally 

contaminated with 3.0 µL cultures of avirulent Leptospira spp 

serovars Hardjo (strain Hardjoprajitno) and Wolffi (strain 3705), and 

virulent Grippothyphosa (strain Moskava V), with 10 6 leptospires 

/mL. Samples were incubated at 30°C for 30 min and frozen in liquid 

nitrogen for 48h. Samples were thawed at room temperature, 

incubated at 30°C for 30 min and cultured in EMJH 30oC for 10 

days. PCR was performed using proteinase K extraction and Lig 1 

and Lig 2 primers. Controls without antibiotic were 100% positive in 

PCR, but only 2/8 and 5/8 cultures for avirulent and virulent strains 

were positive, respectively. From the total of samples, 60/96 

(62.5%) were positive in PCR and 17/97 (17.7%) in culture. A, B or 

C treatments significantly reduced contamination when compared to 

the control; treatment A produced a greater reduction than B, but a 

decrease similar to C. Treatment C was more effective against 

Hardoprajitino and Wolffi than against Grippothyphosa. These 

positive results in PCR or bacteriologic examination confirmed the 

risk of leptospirosis transmission by semen when antibiotic 

treatment is the unique sanitary measure available. 

This paper was supported by FAPESP: Fundaçao de Amparo a 

Pesquisa do Estado de Sao Paulo, Brazil 

Key words: bovine semen, antibiotic extender, Leptospira spp 

899 Looking for a Needle in a Haystack - Identification of Bovine 

Mammary Gland Stem Cells 

M. Kozlowski 1 , M. Gajewska 1 , J. Bierla 1 , B. Gajkowska 2 , T. Motyl 1 

1 



2 



At birth, bovine mammary parenchyma consists of a rudimentary 

duct network. Full mammary epithelium development occurs during 

pregnancy and lactation. The cycle of pregnancy-associated 

mammary gland proliferation, differentiation, and involution occurs 

many times during the lifespan of diary cattle. The development and 

subsequent redevelopment of the mammary gland with each 

successive lactation; involution cycle provides a strong indication 

that a self-renewing population exists within the mammary 

parenchyma. The aim of our study is identification of bovine 

mammary gland progenitor cells using morphological, biochemical 

and molecular markers. Mammary glands were obtained from 2 

years old Holstein heifers. Morphological characteristic of cell in 

mammary gland tissue was performed using electron microscopy 

according to Chepko and Smith designation. Isolation of mammary 

gland epithelial cells was carried out by collagenase/hyaluronidase 

digestion assay. Isolated cells were stained with Hoechst and 

analyzed by FACS to define side population (SP). In the present 

morphological studies we identified small light cells (SLC) similar 

to those previously described in mouse mammary gland. Organelles 

of these cells are small and show no structural evidence of 

specialized function. This may be a combined population of stem 

and primary progenitor cells. Hoechst 3342 efflux is one of the 

features of haematopoetic stem cells. Cells with this ability are 

referred to as the side population (SP). We used this assay to look for 

the side population in bovine mammary gland. We managed to 

identify a small fraction of cells (0,48%) that exhibited a SP 

phenotype. The formation of the side population was blocked by 20 

µM verapamil, which was consistent with previous studies showing 

a requirement for AMB transporter family function in the SP 

phenotype. The results of our study confirmed the existence of a 

small population of stem and progenitor cells in the bovine 

mammary gland. Our group is planning to continue the work on 

further characteristic of these cells in order to create a transcriptomic 

profile of the side population found in bovine mammary gland. 

Key words: mammary gland, stem cells, side population 

900 Comparative Pharmacokinetic Study of Progesterone in Cows 

Following Single Administration of Two Intravaginal Devices 

S. Floc’H 1 , A. Lagarde 2 


2 CEVA santé Animale, Project Department, Libourne, France 

The objective of this study was to compare the kinetic profiles of 

progesterone following single administration of two intravaginal 

devices according to a cross over design. 

Six Prim Holstein non lactating ovariectomised cows were included. 

The study has been carried out according to a single dose, 

randomised, two periods in a cross-over design and a wash-out 

period of 7 days was allowed between each treatment period.The test 

items [A = PRID ® from CEVA S.A.; France and B = CIDR ® Eazi 

Breed from INTER AG; New Zealand] has been administered at D0 

of each period as a single intravaginal insertion which is the 

recommended route for use in bovine females. The devices stayed in 

place for 7 days. Blood specimens were collected at the following 

sampling times for each treatment period: just before the treatment, 

1h, 3h, 6h, 9h, 25h, 33h, 49h, 57h, 73h, 81h, 97h, 105h, 121h, 129h, 

145h, 153h, 169h (just before removal device), 170h, 171h post 

administration. Plasma progesterone concentrations were measured 

by RIA at INRA Nouzilly (France) and described by Saumande J. in 

Steroid Biocem. Molec. Biol. 1991.The main pharmacokinetic 

parameters of plasma progesterone for treatment A and treatment B 

were: Cmax of 4.8 ± 0.7 ng/ml and 3.73 ± 0.6 ng/ml respectively, 

and AUCtot of 666.580 ng.h/ml and 576.3 ng.h/ml respectively. No 

statistically significant difference was noticed for AUCtot 

(p=0.33737) while a statistically significant difference was noticed 

for Cmax (p=0.00599) on behalf of treatment A. A large release of 

progesterone at the beginning of the treatment leads most of the time 

to dominant follicle atresy by a sudden and massive inhibition of 

FSH (Follicle Stimulating Hormone) and LH which are necessary for 

follicle growth and maturation. Therefore the start of a new follicular 

wave can occur allowing the presence of a mature dominant follicle 

at the time of device removal. This prevents persistent dominant 

follicles and therefore allows good fertility at induced oestrus. In the 

present study, the kinetic profiles of progesterone were compared 

following single intravaginal administration of test item A (PRID ® ) 

and test item B (CIDR ® ). Items were administered for a treatment 

period of 7 days according to a single dose. It was concluded that a 

statistically significant difference (p = 0.006) was observed on Cmax 

in favour of PRIDv compared to CIDR ® 

Key words: progesterone, intravaginal device, pharmacokinetic 

901 A Comparison of the Sexual Behaviour of Holstein-Friesian 

Cows between Housed and Pasture Environments 

M. Palmer 1 , G. Olmos 2 , L. Boyle 2 , J. Mee 2 

1 

University of Edinburgh, Royal (Dick) School of Veterinary Studies, 


1 



This experiment compared the sexual ethogram of 46 spring-calved 

cows in two management environments [cubicle housing with a 

total mixed ration (HOUSED) and rotational pasture with 

concentrate supplementation (GRASS)]. Cows were blocked and 

randomly allocated to treatment and were monitored by one 

observer from ten days postcalving for nine weeks on the same 

farm. The occurrence of nine behaviours (ano-genital sniffing given 

or received, chin resting given or received, head-to-head butting, 

mounting attempts, evasion of mounting attempts, mounting and 

standing to be mounted) was recorded during the thrice daily (6.00, 

12.00 and 19.00h) 20 minute visual observation sessions. Milk 

sampling for progesterone analysis (EIA) was carried out on 

Mondays, Wednesdays and Fridays to determine the dates of true 

standing oestrus. Data in the 24h before and after standing oestrus, 

(seven peri-oestral observation periods), were analysed by proc 

FREQUENCY, GENMOD, NPAR1WAY, TTEST and 

UNIVARIATE, as appropriate, in SAS. In both treatment groups 

Reproduction and Biotechnology 203

the median number of attempted mounts, ano-genital sniffs given, 

chin resting given or received and head-to-head butting changed 

over the 48h peri-oestral period peaking during the standing oestrus 

observation session (P0.05). During the standing 

oestrus session, GRASS cows stood to be mounted more frequently 

than the HOUSED cows (median, Q1, Q3: 2.5,1,3 and 0.0,0,1, 

respectively, P0.05). These results 

demonstrate that HOUSED cows do not show the same peri-oestral 

increase in mounting behaviour characteristic of GRASS cows. The 

lower libido of HOUSED cows was attributed to slippery underfoot 

conditions, more lameness, the stress of confinement and possible 

nutritional interactions. These results have implications for oestrous 

detection systems which rely on mounting behaviour alone. 

902 The Effect of 2 Commercial GnRH Products at the Same 

Dose on the Pituitary LH Release in Dairy Cows 

F. Deletang 

19 Rue Claude MASSE, Animal Reproduction, La Rochelle, France 

The objective of this study was to evaluate the LH secretion 

response following administration of a 100 mcg dose of two 

different formulations of gonadorelin (Product A : 

Cystoreline ® /Ovarelin ® = gonadorelin diacetate tetrahydrate, CEVA 

S.A., France and product B : Fertagyl ® = gonadorelin acetate, 

JANSSEN, Belgium). Despite having the same active ingredient, 

the two products present two different salt forms and dosage 

recommendations (100 mcg vs. 250 to 500 mcg respectively). Six 

Prim Holstein lactating cows were studied over 2 consecutive 

reproductive cycles. Cows were individually included the day of an 

apparent oestrus, noted D0 of cycle I and were randomly allocated 

to sequenced treatment AB or BA. The two products were 

administered as a single intramuscular injection of 100 mcg of 

active ingredient corresponding to 2ml of product A and 1ml of 

product B. Product A or B was given at D11 of the cycles I and II 

according to a cross over design. Blood was collected for 

determination of LH by ELISA (REPROKIT ® ) at the following 

times : T-1h, T-0.5h and T0h before treatment and T0.25h, T0.5h, 

T0.75h, T1h, T1.25h, T1.5h, T1.75h, T2h, T2.25h, T2.5h, T2.75h, 

T3h, T3.5h, T4h, T5h, T6h after treatment. A maximum peak of 

plasma LH was clearly identified approximately 2 hours post GnRH 

injection. A delay of 6 hours was observed for plasma LH 

concentration to return to basal values as before gonadorelin 

treatment. The main pharmacokinetic parameters observed Cmax 

and AUCtot were computed using the Log Linear Method. 

Differences were observed between A and B with a Cmax of 10.5 ± 

5.9 mcg/l and 7.5 ± 3.3 mcg/l respectively and an AUCtot of 82.1 ± 

74.5 mcg.h/l and 46.9 ± 30.3 mcg.h/l respectively. For treatment of 

cystic cows 100 mcg is the optimal gonadorelin dose, it is the one 

able to trigger a LH release similar to that of a normal LH surge in 

healthy cyclic cows during the luteal phase.Cystoreline ® /Ovarelin ® 

appeared to induce a higher LH peak and a more total LH release 

than the other product, but these differences were not significant (p 

> 0.05). Our results tie up with other comparative studies, Martinez 

et al. (Can. Vet. J. Vol. 44 - 2003) observed a LH peak response of 

6.6 ng/ml after Cystoreline ® and 4.7 ng/ml after Fertagyl ® 5 to 7 

days after ovulation. This suggested that the formulation and the 

salt form directly impact bioavailability of the product. 

Key words: GnRH, formulation, pharmacokinetic 

903 Economic Decision for Time of Cystic Ovarian Disease 

Treatment in Dairy Cows 

M. Farhoodi, K. Valipoor 

Islamic Azad University, Science & Research Branch, Faculty of 

Veterinary Medicine, Tehran, Iran 

Introduction: Cystic ovarian disease (COD) is a major reproductive 

disorder of dairy cattle. COD significantly prolongs the time 

postpartum to first estrus, extends days open and, therefore, the calving 

interval and leads to economic losses. The objectives of the present 

study were to evaluate the profitability of treating cows in the early 

postpartum period. 


Material and Methods: The study was conducted in a large dairy 

herd of 720 Holstein cows in Tehran. In this herd all cows were 

examined 30 days postpartum and cows, which had fluid-filled 

structures greater than 2.5 cm in diameter on one or both ovaries in 

rectal examination were determined as cystic cows. Cystic cows 

were divided in two treatment groups. Some cows were treated at the 

time of diagnosis (day 30 postpartum), (early treated group), (n=86) 

the others 12 days later if they did not recover (late treated group), 

(n=51) with GnRH and further 10 days later with PGF 2a 

(dicloprostenol). In each group, days open, days to first service, 

service per conception, percent of pregnancy before 120 days post 

calving and direct costs including those of the veterinarian, the drugs 

and the labor were estimated. Data were analyzed using Student t 

test. Direct and indirect costs were measured and added together. 

Then difference of economic losses has been estimated in two 

treatment groups. 

Results: Days open and days to first service in the early treated 

group (75.74±43.36 and 53.86±14.26) were better than in the late 

treated group (96.14±42.82 and 65.78±19.12)(P

905 Comparison of Different Techniques to Evaluate Membrane 

and Acrosome Integrity of Bovine Cryopreserved Sperm 

Cells 

J. Carvalho 1,2 , G. Machado 1,2 , M. Franco 1,2 , R. Rumpf 1,2 , 

M. Dode 1,2 

1 Embrapa Recursos Genéticos e Biotecnologia, Laboratório de 

Reproduçio Animal, Brasília, Brazil 

2 Universidade de Brasília, FAV, Brasília, Brazil 

During fertilization process, only spermatozoa with intact plasma 

membrane are able to go through capacitation, to bind to the zone 

pellucida, to undergo acrosome reaction and to fertilize an oocyte. 

Since membrane and acrosome integrity are essential for the initial 

steps of fertilization, their evaluation should be use as a routine 

tests. Stain exclusion methods had been used to assess those 

features. With the development of the staining technology the 

combination of fluorescent stains became widely used, however 

those techniques require fluorescence microscopy, which is not 

always available. The objective of the present study was to compare 

techniques to evaluate membrane and acrosome integrity of bovine 

sperm cells, using or not fluorescent probes. To evaluate membrane 

integrity eosin-nigrosin (E/N) and a combination of 

carboxyfluorescein diacetate and propidiun iodide (CFDA) stains 

were used. For acrossome reaction, dual stain using trypan blue and 

giensa (TBG) and the combination of peanut agglutinin conjugated 

with fluorescein isothiocyanate (PNA-FITC) were performed. 

Frozen semen from four different bulls was used in a three replica 

experiment. In each replicate three straws were thawed and were 

pooled in a centrifuge tube. From that pool, samples were removed 

for evaluation of membrane and acrosome integrity using bright 

field (EN and TBG) and fluorescence (CFDA and PNA-FITC) 

microscopy, in which 200 cells per slide were evaluated. Data were 

compared using a t test. The mean percentage of spermatozoa with 

intact plasma membrane was similar (P>0.05) for the EN and CFDA 

technique, being 53.41 ± 14.57% and 45.39 ± 7.46%, respectively. 

However, a higher (P

909 The effect of “GnRH” and “ PGF2a” Treatment at the Time 

of Artificial Insemination on Conception Rate of 

Synchronized Dairy Cow 

M. Asfia 1 , M. Farhoodi 1 , A. Bahonar 2 

1 Faculty of Veterinary Medicine Islamic Azad University Karaj, 

Animal Reproduction Gynecology & Obstetrics, Tehran, Iran 



Acquiring higher conception rate (CR) via AI is one of the major 

issues in dairy cow reproduction. In this study, the effect of IM 

treatment with GnRH and its combination with PGF 2a at the time of 

AI on CR, during the heat stress period was evaluated in two groups 

of Holstein dairy cows with good BCS (2.5-3.5). Our cows were 

synchronized with a single injection of PG if a corpus luteum was 

confirmed by rectal palpation and ultrasound examination, then AI 

was performed 80 (3 days, n=114) or 96 hrs (4 days, n=103) later for 

cows that had good mucosal discharge and uterine turgidity. Tow 

protocols were administrated at AI:1) 0.01 mg of GnRH (buserelin), 

2) 0.01 mg of GnRH + 500 ag PGF 2a (cloprostenol). Analysis of 

variance was used. In the 1st group (80 hrs AI) we had 46 controls, 

35 treated with “GnRH” and 33 treated with “GnRH+ PGF 2a”.GnRH 

protocol awarded the best CR in compare with “ GnRH+ PG 2a” 

protocol and control group (primiparous CR 50%,56%,45% , 

multiparous CR 45%, 35%, 42% and all cows CR 46%,45%,43%). 

Even in the 2nd group (96 hrs AI) we had 54 controls, 27 treated with 

“GnRH” and 22 treated with “GnRH+ PGF 2a”, the best CR were 

related to GnRH protocol (primiparous CR 45%, 11%, 23%, 

multiparous CR 44%, 38%, 41% and all cows CR 44%, 27%, 33%). 

CR of primiparous cows with 80 hrs AI and “GnRH” treatment was 

better than primiparous cows with 96 hrs AI and “GnRH+PGF 2a” 

treatment (P=0.06). Also CR of 80 hrs AI with “GnRH+PGF 2a” was 

better than 96 hrs inseminated cows with “GnRH+PGF 2a” (P=0.07). 

this study indicates that GnRH and PGF 2a treatment at AI may 

increase CR in 80hrs AI primiparous cows in heat stress situation. It 

is considerable that 80 hrs AI group had higher CR in compare with 

controls which inseminated at heat (46% vs. 35%). 

910 Influence of Plasmatic Membrane Cryoresistence on Post- 

Thawed Spermatic Motility in Frozen Semen of Santa Ines 

Rams 

H. Azevedo 1 , S. Bicudo 2 , M. Maia 3 , D. Sousa 2 , L. Rodello 2 , 

C. Sicherle 2 

1 Embrapa Coastal Tablelands, Aracaju, Brazil 

2 Sao Paulo State University - UNESP, Faculty of Veterinary 

Medicine and Animal Science, Botucatu, Brazil 

3 Embrapa Tropical Semi-Arid, Petrolina, Brazil 

The semen of 25 Santa In’s rams was cooled, frozen, thawed and 

incubated in an oviduct synthetic medium and evaluated as for 

subjective motility (MOT, 0-100%) in moments (MOMENT): fresh 

(FR-S), refrigerated (RE-S), frozen-thawed (FT-S), incubated (IN- 

S). Based on the plasmatic membrane integrity (PMI, X=22.1%) 

measured in FT-S by association of propidium iodide (PI) and Pisum 

sativum agglutinin conjugated with fluorescein isothiocyanate (PSA- 

FITC), rams were grouped in three cryoresistence levels (CRYO): 1: 

inferior (X 10.3%); 2: intermediate (10.3%>X 29.9%); 3: superior 

(X>29.9%). It was used the variance analysis (ANOVA) in a 

completely randomized design and Tukey method to compare the 

averages (P

female.The accuracy of ultrasonographic fetal sex determination was 

confirm after birth of calves. The accuracy of female fetal sexing in 

group A was 87.2% and of male 88.5%, total 87.7%. The accuracy of 

female fetal sexing in group B was 86.3% and of male 89.5%, total 

87.6%. We did not find differences in the accuracy of fetal sex 

determination betwen groups A and B. The better results were in 

male fetal determination. The optimal days for fetal sex 

determination were 55 to 65 days of gestation. After 90 days of 

gestation there were problems associated with obtaining an adequate 

view in some cows. 

Funded by 1/3484/06 and AV 4/0009/07. 

Key words: dairy cow, fetal sex, ultrasound 

913 In vitro Cultivation of Anaplasma Marginale in two Bovine 

Endothelial Cell Lines 

G.S. Luna-Castro 2 , C.A. Vega y Murguía 1 , 

S.D. Rodríguez-Camarillo 1 

1 

Cenid-Pavet, INIFAP, Anaplasmosis, Jiutepec, Morelos., Mexico 

2 

Facultad de Medicina Veterinaria y Zootecnia - UNAM, México 

D.F., Mexico 

Anaplasma marginale, a Gram-negative intracellular bacteria that 

parasitizes bovine erythrocytes, is classified in the order 

Rickettsiales, family Anaplasmataceae, and causes bovine 

anaplasmosis. This disease is characterized by progressive anemia, 

fever, decrease in body weight and death, causing considerable 

economic losses. For its propagation requires mechanical or 

biological vectors. At present, there is not an effective vaccine 

commercially available for Mexican field conditions. It has been 

raised that endothelial cells may play an important role as key 

element in the development cycle of this microorganism within the 

bovine host. The present work attempted to establish the in vitro coculture 

of A. marginale in two bovine endothelial cell lines along its 

typical host cell, the bovine erythrocyte. A. marginale Yucatan 

isolate was multiplied in susceptible splenectomized donors and 

infected erythrocytes (ie) were used as source of infectious material 

for cultures. The cell lines, BCE C-D/1b from cornea and BUVEC 

E 6E 7, of umbilical cord, were used. Cultures were maintained in 24well 

plates. Each 16 mm diameter well contained a treated cover slip 

in the bottom. The initial seeding dose was approximately 1 x 10 3 

cells per mm 2 in 1,000 µl and plates were incubated at 37 °C in a 5% 

CO 2/air, atmosphere. Every 24 hours, 1,000 Ïl media /well were 

exchanged. Once monolayer was formed, initially 1,250 µl of a 50% 

ie suspension were added to each well and periodical media 

exchange continued; eventually, non-infected bovine erythrocytes 

were added to cultures. Giemsa stained smears to monitor infection 

were prepared a time of exchanging media, by sampling sedimented 

cells. For each pass, resuspended cells without medium exchange 

were transferred to a newly formed monolayer in the next well, 

which also contained a treated cover slip and cover slips from 

previous well were recovered for microscopic observation. Cultures 

remained positive during the 151 day trial period, but no differences 

were noticed among cell lines due to great fluctuation in the number 

of observed infected erythrocytes. 

Partially supported by: CONACyT-SAGARPA grant 2004-C01-71 

Key words: bovine, in vitro culture, Anaplasma marginale 

914 Epidemiological Data of Postpartum Dairy Cows with Acute 

Metritis 

B. Huyghe 1 , Sa, de Carlos 1 , P. Cox 1 , J. Swinkels 2 

1 Intervet Schering-Plough, R&D, Angers, France 

2 Intervet Schering-Plough, Int. Marketing, Boxmeer, Netherlands 

Objectives: The use of cefquinome for intra-uterine treatment of 

acute metritis was tested in four field trials between 2001 and 2006 

in France, Hungary, Germany and the Netherlands. The objective of 

these trials was to determine a cefquinome dose adequate for intrauterine 

treatment of acute metritis in cattle and to evaluate the 

efficacy of the determined dose as compared to positive control. 

Data from the anamnesis of animals included in these studies were 

analyzed and are presented here. 

Materials and Methods: A total number of 1060 cows (4-14d 

postpartum) and diagnosed with acute metritis were included in the 

studies. Acute metritis was defined as the presence of red-brown 

watery, foul smelling uterine discharge. Cows recently treated with 

intra-uterine or parenteral antibiotics between calving and the 

diagnoses were excluded. Inclusion data from the cows were 

compiled and descriptive statistic performed. 

Results: Parity distribution among the cows included in the study was 

as follows: 46% primiparous, 22% parity 2, 15% parity 3 and 9% 

parity 4. The distribution of the day of acute metritis diagnosis in 

respect to calving followed a Gauss shaped curve, centered on day 7 

with 70% of the cases included between day 6 and day 10 postpartum. 

Majority of animals (52.5%) experienced normal calving (nonassisted) 

preceding their acute metritis episode, and 74% had no 

retained placenta. General health was not altered in 61% of cases, 

while moderate health alteration (reduced activity, no reduction in 

milk production or feed consumption) was recorded for 36%. This is 

consistent with the rectal temperature measurements recorded during 

these studies, with 89% of cases showing rectal temperature < 39.5°C 

at inclusion. Average rectal temperature at inclusion was slightly 

above normal (38.5°C) at 38.8°C ± 0.55. 

Conclusions: Although assistance at calving and retained placenta are 

known risk factors for acute metritis, the majority of the animals 

diagnosed with acute metritis were normal for these parameters. In the 

described field studies, we have observed that general clinical signs 

(altered general health and fever) associated with acute metritis had 

marginal presence. 

915 A Field Efficacy Study on the use of Cobactan 2,5% in Cows 

for the Treatment of Acute Puerperal Metritis 

E. Niro 1 , J. Swinkels 2 

1 


Unit, Milan, Italy 

2 

Intervet Schering-Plough, Int. Marketing, Boxmeer, Netherlands 

Objectives: The objective of the study was to evaluate the efficacy of 

cefquinome, administered parenterally in the treatment of acute 

puerperal metritis in cows. 

Materials and Methods: A total number of 97 Holstein-Friesian cows 

(

3 

COOPEVOLIA, Centre Zootechnique, Verdun sur le Doubs, France 

4 


5 



6 


Objectives: Estrus synchronization method based on Crestar SO ® 

foresees a single artificial insemination (AI) performed without estrus 

detection at 48h after the removal of norgestomet implant in all animal 

types. The aim of this experiment was to estimate the influence of 

earlier insemination timing on the pregnancy rate (PR) following the 

induced estrus in Charolais heifers. 

Materials and Methods: A total number of 252 Charolais heifers, 

housed in 14 farms were included in the study. The animals were 

assigned to 2 groups: 36h AI and 48h AI, matched by age, body 

condition score and body weight. Estrus synchronization was 

performed according to the following treatment schedules: injection 

of 10mcg buserelin (Receptal ® , Intervet) with simultaneous 

placement of subcutaneous norgestomet implant (Crestar SO ® , 

Intervet) in the ear auricle. In 48hAI group the implant was removed 

after 9 days and 36h before implant removal PGF 2a was 

administered. In 36h AI group norgestomet implant was removed 

after 9.5 days and PGF 2a given at 48h before. All animals were then 

artificially inseminated at 36h or 48h post implant removal for 36h 

AI and 48h AI groups respectively. Pregnancy diagnosis was 

performed with ultrasound at 51.1± 2.3 days post AI. Statistical 

significance was assessed with multivariable analysis of logistic 

regression. 

Results: No statistically significant difference was found in the average 

PR after single AI at the induced estrus for insemination at 36h and 48h 

after Crestar SO ® removal (PR/36h±61.9%; PR/48h-58.8%). Marked 

differences were noted between fertility results reported for individual 

different farms included in the study. 

Conclusions: Hastening of insemination time from 48h after 

norgestomet implant removal to 36h had no negative influence on PR 

achieved after a single AI at the induced heat. This change may offer an 

attractive practical alternative for service in heifers of good body 

condition synchronized with Crestar SO ® . 

917 Influence of Number of Inseminations on Pregnancy Rates 

Achieved at Estrus Induced with Crestar SOR in Female 

Charolais Cattle 

C. Ponsart 1 , B. Grimard 2 , P. Humblot 1 , M. Dewaele 3 , 

M. Driancourt 4 , R. Fournier 5 , M. Ptaszynska 6 , V. Rosso 5 

1 

UNCEIA, Departement R&D, Maisons-Alfort, France 

2 

UMR INRA/ENVA, Departement R&D, Maisons-Alfort, France 

3 

CECNA, Research, Migennes, France 

4 


5 



6 


Objectives: Estrus synchronization method based on Crestar SO ® 

foresees a single artificial insemination (AI) performed without estrus 

detection at 48h after the removal of norgestomet implant in all animal 

types. The aim of this experiment was to estimate the influence of the 

number of inseminations performed (1AI vs. 2 AIs) on pregnancy rates 

in lactating Charolais cows. 

Materials and Methods: A total number of 266 cows (64 

primiparous, 202 multiparous) maintained in 19 farms were included 

in the study. Estrus synchronization and insemination was performed 

between January and March 2006. The cows were divided into 2 

treatment groups matched by parity and body condition score: 1AI 

group (n=132) and 2AI (n=134). The synchronization schedule 

included an injection of 10mcg buserelin (Receptal ® , Intervet) with 

simultaneous insertion of norgestomet implant (Crestar SO®, 

Intervet), followed by an injection of PGF 2a on D7 and implant 

removal 48h later. At implant removal PMSG was given (500- 

600IU, Folligon ® , Intervet). The cows in 1AI group were 

inseminated once at 48h after the implant removal. Group 2AI was 

inseminated twice at 24h and 48h after the implant removal. 

Pregnancy diagnosis was performed with ultrasound at 44.1+/- 6 

days post AI. Statistical significance was assessed with 

multivariable analysis of logistic regression. 

Results: Additional insemination performed at 24h after implant 

removal brought no improvement in pregnancy rates (PR/1AI-66.7% 


vs. PR/2AI-60.5%). In primiparous cows single AI gave significantly 

better results than 2 services (PR/1AI-78.6% vs. PR/2AI-44.4%), while 

in multiparous group no statistical difference was found (PR/1AI- 

63.5% vs. PR/2AI-66.3%). 

Conclusions: The results of this trial confirmed that a single AI 

performed at 48h after the removal of Crestar SO ® implant is sufficient 

to achieve satisfactory pregnancy rates. The negative effect of 2AIs on 

pregnancy rates in primiparous cows remains to be confirmed on a 

larger set of animals. 

918 Comparison of Fertility Results with two Progestin based 

Synchronization Methods in French Conditions 

B. Grimard 1 , C. Ponsart 2 , P. Humblot 2 , C. Gipoulou 3 , 

V. Rosso 4 , R. Fournier 4 , M. Ptaszynska 6 , M. Driancourt 5 

1 Ecole Nationale Veterinaire d’Alfort, UP Zootechnie, Maisons- 

Alfort, France 

2 UNCEIA, UP Zootechnie, Mansoura, France 

3 Clinique Vétérinaire, Clinique, Castres, France 

4 Intervet Schering-Plough, Farm and Production Animals Business 


5 Intervet Schering-Plough, R&D, Angers, France 


Objectives: The aim of this study was to evaluate the efficacy of a new 

Crestar SOv method followed by a single fixed time AI as compared to 

Crestarv and Crestar SO ® methods based on different mechanisms that 

prevent the persistent dominant follicles formation: 

estradiol+norgestomet induce atresia of large follicles present on the 

ovaries at the start of treatment, while the use of buserelin at implant 

insertion synchronizes follicular waves and triggers ovulation of LHdependent 

follicles. 

Materials and Methods: The study included 3 experiments 

conducted between 2002 and 2005 according to the following 

protocol: Study 1: 382 Charolais heifers and cows (cyclicity 51%), 

estrus was synchronized with Crestar ® (Day 0-norgestomet implant 

and injection of 3mg norgestomet+5mg estradiol valerate) or Crestar 

SO ® (Day 0- norgestomet implant +10mcg buserelin (Receptal ® , 

Intervet); Day 8 “PGF 2a”(15mg luprostiol; Prosolvin ® , 

Intervet) Day 10 implant removal + 500-600IU PMSG (Folligon ® , 

Intervet), followed by AI at 48h (heifers in Crestar ® and all females 

in Crestar SO ® group) or 56h (cows in Crestar ® group) after implant 

removal. Pregnancy rate was determined with ultrasound at 30-50d 

after AI. Study 2: in 139 cows (98 Limousine, 41 Blonde 

d’Aquitaine; cyclicity 73%), estrus was synchronized as in the Study 

1, with 500IU PMSG dose. Pregnancy rate was determined with 

PSPB assay at 45d after AI. Study 3: in 327 cows (134 

Montbeliardes, 99 Normandes, 94 Prim’ Holstein), estrus was 

synchronized as in Study 1, with a simple dose of PGF 2a (15mg 

luprostiol; Prosolvin ® , Intervet) 48h before the implant removal and 

400IU PMSG dose. Pregnancy rate was evaluated with ultrasound or 

PSPB assay at 42-48d after AI. 

Results: Pregnancy rates in the three studies were as follows: Study 1: 

PR/Crestar-46% vs. PR/Crestar SO-51%; Study 2: PR/Crestar ® 73%vs. 

PR/Crestar SO-75%; Study 3: PR/Crestar-46% vs. PR/Crestar SOR-44%. 

Conclusions: Fertility after the synchronization of estrus with Crestar ® 

and Crestar SO ® proved to be similar in evaluated breeds and age 

groups. 

919 Effect of the Type of Semen (sexed vs. non-sexed) and Time of 

AI on Pregnancy Rates in Nelore Cows Inseminated at Fixed 

Time 

A. Souza 1 , L. Mundim 2 , P. Venturin 3 , H. Ayres 1 , L. Gimenes 1 , 

J. Sales 1 , C. Martins 1 , P. Baruselli 1 

1 

FMVZ/USP, Departamento de Reproducao Animal, Sâo Paulo, 

Brazil 

2 

Veterinary Clinic, Departamento de Reproducao Animal, Botucatu, 

Brazil 

3 

Fazenda Sao Luis, Gerente Administrativo, Cascavel, Brazil 

Objectives: The objective of this study was to evaluate the effect of the 

reduction in the interval between AI and ovulation on pregnancy rates 

in Nelore cows inseminated at fixed time with either sexed or regular 

(non-sexed) semen. 

Materials and Methods: The experiment was performed in 3 

commercial farms in Brazil. On D0 animals (n=389) received 2 mg

estradiol benzoate (Estrogin ® , Farmavet), a norgestomet ear implant 

(Crestar ® , Intervet) and 0.150mg D-cloprostenol (PGF2a, 

Prelobanv, Intervet) followed on Day 10 (48 h post implant removal) 

with GnRH (100 µg, Fertagyl ® , Intervet). The cows were then 

divided into 4 experimental groups: C54h: AI/regular semen 

(20x106 sperm/dose; n=95) at 54h after Crestar withdrawal; S54h: 

AI/sexed semen (2.1x106 sperm/dose; n=99) at 54h; C60h: 

AI/regular semen (20x106 sperm/dose; n=98) at 60h; S60h: 

AI/sexed semen (2.1x106 sperm/dose; n=97) at 60h. Semen of a 

single Nelore bull was used to inseminate all cows. Sexed semen (Y 

and X) was used in farms 1 and 2, but only X-semen in farm 3. 

Pregnancy rates (PR) were assessed 30d post AI with ultrasound 

(Easi-Scan®, BCF technology). On farm 3 fetal sexing was 

performed 60 days post AI. Comparison of pregnancy rates between 

treatments was performed with proc GLIMMIX do SAS. 

Results: The PR were: C54H 48,4%, S54H 37,4%, C60H 55,1%, 

and S60H 46,4%, with the type of semen clearly affecting pregnancy 

rates (Regular: 51,8% vs. Sexed: 41,8%). However, the results 

achieved with sexed semen at 60h were similar to regular semen at 

either 54 or 60h. There was no effect of time of AI (54hPR-42,8% vs. 

60hPR-50.8%). Fertility was similar in animals inseminated with Xsemen 

(52,0%) and Y-semen (49,0%). For both insemination times, 

the use of sexed semen produced similar proportion of the desired 

sex (S54h: 92,9% vs. S60h: 94,7%). 

Conclusions: Pregnancy rates in postpartum Zebu cattle inseminated 

with sexed semen at a fixed time are acceptable and statistically similar 

to those achieved with regular semen when the timed AI is 60h after 

implant removal. 

920 The First Report of Bovine Amorphus Fetus in a Triplet 

Pregnancy 

H. Nourani 1 , A. Shirazi 2 

1 Department of Pathobiology, School of Veterinary Medicine, 

Shahrekord University, Iran 

2 Research Institute of Animal Embryo Technology, Shahrekord 

University, Shahrekord, Iran 

Amorphus fetus is a rare and incidental finding in bovine placentas 

especially. This anomaly has been reported in bovine twin 

pregnancies. Amorphus fetus also called acardiac monster, 

holoacardius amorphus or acardius amorphus, amorphus globosus 

and fetal mole. These terms are used to describe the acardius 

anomaly when the general body form was not recognizable. A six 

years old cow was referred to Veterinary clinic of Shahrekord 

University due to fetal membranes retention in May 2007. An 

amorphus fetus was expelled following the delivery of two normal 

calves. The amorphus fetus, weighing 30 gram was kidney-shaped 

with two convex and concave surfaces and borders. The convex 

surface is composed of three segments but the other one had two 

parts. The entire mass was covered with hair, which bore the 

characteristic pigmentation of Holstein cattle. Histological 

examination of Amorphus fetus revealed that this mass was covered 

by thin keratinized squamous epithelium and collagenous connective 

tissue, hair follicles and associated sebaceous glands were seen 

beneath the epidermis. In addition to collagenous connective tissue, 

skeletal muscle and adipose tissue were observed in the central part 

of the mass. The purpose of this case report is to describe 

macroscopic and microscopic features of the amorphus fetus and to 

increase the practitioner,s awareness about it. To our knowledge, 

there is no report about occurrence of amorphus fetus along with two 

normal calves in cattle and, for the first time, it is reported in this 

study. 

Key words: Amorphus fetus, Acardius, morphology, calf 

921 Oocyte Production by Ovum Pick up is Influenced by the 

Paternal Lineage of Donor Cow and Season 

R. Calegari 1 , A. Martins Jrv, C. Silva 1 , A. Crespilhov 

1 Sao Paulo State University - UNESP, Clinical Surgery and Animal 

Reproduction, Aracatuba, Brazil 

2 Sao Paulo State University - UNESP, Department of Animal 

Reproduction, Botucatu, Brazil 

Our previous report has shown that the maternal lineage of oocyte 

donor affects the number and quality of recovered oocytes obtained 

by ovum pick up in Nellore cows. The aim of the present study was 

to investigate the effect of paternal lineage of oocyte donor and 

season on oocyte production in an OPU program. Thus, two welldefined 

lineages (Karvadi and Tal Mahal) of Nellore (Bos indicus) 

cows, referred to as KD and TM, respectively, were subjected to 

ultrasound-guided follicular aspirations at random stages of the 

estrous cycle over a period of two years. Follicles were aspirated 

using a real time B-mode scanner (Aloka SSD-500, Japan) and a 5 

MHz transvaginal convex transducer equipped with a needle guide 

attached a single lumen needle. Then, all visible follicles were 

punctured and harvested oocytes were counted, examined 

morphologically and classified into seven categories according to 

the cumulus cell investment and their cytoplasmic appearance. Data 

of the numbers of oocytes retrieved per session were analyzed using 

Tukey’s test with P

923 Production of Bovine Blackleg Vaccine by Fermenter 

R. Pilehchian, M. Moosawi, A. Jabari 

Razi Research Institute, Anaerobic Vaccine Production, Karaj, Iran 

Blackleg is an actual and fatal disease of cattle caused by Clostridium 

chauvouei. The object of this study was to prepare a potent vaccine 

against blackleg of cattle. Attempts were made to prepare and formulate 

the ingredients in order to obtain high yield of Clostridium chauvoei 

organisms in culture medium by fermenter. The medium consisted of 

efficient amount of nitrogen and carbon sources. A large-scale vaccine 

with satisfactory results was obtained in fermenter. More, than 

1,450,000,000 organisms/ml was obtained by the end of incubation 

period. The safety and potency of the prepared vaccine was determined in 

Guinea pigs and Sheep according to British pharmacopoeia (veterinary) 

with satisfactory results. In a preliminary experimental trial, 9 calves were 

divided into 3 groups. 2 groups were immunized by 2 doses of the 

vaccine, with an interval of 15 days, and then challenged in 2 weeks after 

the 2 nd injection. The third group was considered as a non-immunized 

control. According to the results, the vaccine prepared an adequate 

protection against the wild strain of Clostridium chauvouei. Animals of 

the control group were rapidly died after injection of the challenge strain. 

Key words: Bovine Blackleg vaccine, fermenter 

924 Study on Production of Reduced Dose Blackleg Concentrated 

Vaccine 

M. Moosawi, R. Pilehchian, A. Jabari, L. Abdolmohamadi 

Razi Research Institute, Anaerobic Vaccine Production, Karaj, Iran 

Blackleg is generally known to affect cattle but sheep, goats, swine, 

camels, deer and mink are also susceptible. Blackleg in cattle has been 

recognized in Iran. Formalized whole culture vaccine have been 

prepared for immunization of cattle and sheep against the disease at 

Razi Institute using traditional manner and also by fermenter. The main 

object of this study was to prepare and formulate a reduced dose potent 

vaccine for immunization of cattle against blackleg in Iran. There are a 

few references in the literature concerning production of blackleg 

vaccine by fermenter. After an active growth duration, detoxification 

and killing of bacteria was done by adding formaldehyde (37%) at the 

rate of 0.7% and pH of culture adjusted to 7.Aluminum hydroxide gel as 

adjuvant was used for the Clostridium chauvoei vaccine in sufficient 

quantity to give 10 percent gel in final concentration. The concentrated 

vaccine was mixed by slowly stirring for 30 minutes at 20 °C.The 

quality control of the experimental vaccine was determined according to 

the European pharmacoepia Veterinary 5 th edition (2004). Three healthy 

cattle were selected for safety test of the vaccine. Each cattle was 

injected subcutaneously 2ml of the vaccine and observed for ten days. 

Ten healthy guinea pigs (300-400gr) were injected subcutaneously with 

2ml of the vaccine as primary dose, followed by three weeks later a 

second injection of the same quantity of the vaccine as the secondary 

dose. After two weeks the vaccinated and unvaccinated guinea-pigs 

were injected intramuscularly with 4 MLD for vaccinated and 1MLD 

for unvaccinated of 24 hrs virulent culture of Clostridium chauvoei 

respectively. Results shows that the routine blackleg vaccine with 3ml 

dose gave good immunity in injected animals but the new experimental 

concentrated vaccine with reduced dose of 2ml has given highly 

immunity in guinea pigs and cattle. The reduced dose of vaccine could 

be replaced of the routine blackleg vaccine because of long immunity in 

vaccinated animals and also because of economic points of view. 

Key words: Blackleg vaccine, cattle, fermenter, pharmacoepia 

925 Bovine Embryo Development after In vitro Maturation with 

Glycine 

C. Silva, A. Martins Jr, R. Calegari 

Sao Paulo State University - UNESP, Clinic, Surgery and Animal 

Reproduction, Aracatuba SP, Brazil 

The purpose of this study was to investigate the effects of glycine during 

in vitro maturation of bovine oocytes on their subsequent embryonic 

development after IVF. Oocytes were aspirated from follicles (2 to 6 mm) 

of ovaries obtained from slaughtered cows. Oocytes having at least 3-4 

layers of cumulus cells and homogeneous cytoplasm were washed in 

TCM-199 with 2.2 mg/ml sodium bicarbonate, 0.02 mg/ml sodium 

pyruvate, 0.05 mg/ml penicillin, 1 mg/ml PVA, and 2.38 mg/ml HEPES. 


Groups of 15 to 25 oocytes were matured for 24 h in 100 µl drops of TCM 

(Gibco ® , USA) with 0.5 µg/ml FSH (Pluset ® , Spain), 50 Ïg/ml LH 

(Lutropin-V ® , Canada), and 10% FCS (Nutricell, Brazil), supplemented 

with glycine, according to the experimental groups: I (0,67mM; n=144), 

II (10 mM; n=132), and III (100 mM; n=130). For inseminations, frozenthawed 

sperm was obtained by density gradient separation (Percoll). 

Following a centrifugation (700 x g) for 30 min, the resulting pellet was 

washed in TALP by centrifuging 200 x g for 5 min. Afterwards, the pellet 

was re-suspended to 50 µl with TALP plus PHE and heparin. Gametes 

were co-incubated for 20 h for IVF. Then, cumulus cells were removed 

and 15 to 20 presumptive zygotes were cultured in 50 µl drops of culture 

medium (m-SOF) up to day 8 post-insemination (pi). Oocytes and 

embryos were incubated at 39 °C under a humidified atmosphere of 5% 

CO 2 in air. Embryonic development was monitored, and 50% of the 

culture volume was replaced with fresh medium at 72 and 120 h pi. 

Percentages of cleaved oocytes and oocytes reaching morula 

(M)/blastocyst (B), blastocyst/expanded blastocyst (EB), and expanded 

blastocyst/hatched blastocyst (HB) stages were recorded at 72, 144, 168, 

and 192 h pi, respectively. Data of five replicates were analyzed using 

ANOVA and Bonferroni t-test. Differences of P

Conclusion: the results of the present study indicated that the 

incidence of genital tract abnormalities/diseases between the 

slaughtered cows were relatively high (12.6%). Moreover, an 

outstanding number of the pregnant cows were slaughtered since the 

lack of early pregnancy detection. 

927 Bovine Embryo Fertility Rates Comparision in Morula and 

Blastocyst Stage, with the Frozen Embryos Transfer 

Technique in Field Conditions with a Manual Device 

P. Cano-Celada 1 , J. Avila-Garcia 1 , L. Ocampo-Camperosv, 

J. Valencia-Mendez 1 

1 Universidad Nacional Autónoma de México,Facultad de Medicina 

Veterinaria y Zootecnia, Producción Animal:Rumiantes, Mexico, 

DF., Mexico 

2 Universidad Nacional Autónoma de México,Facultad de Medicina 

Veterinaria y Zootecnia, Animal Medicin, México D.F., Mexico 

The goal of this paper was to compare the bovine embryo fertility 

rates in morula and blastocyst stage frozen in field conditions with a 

manual device. The research was carried out in Casa Blanca ranch, in 

Acatzingo, Puebla, Mexico. 387 transferences were made from 

Black swiss donor cows embryos. The embryo collection was made 

by the non surgical method. Immediatly after the morfological 

evaluation, embryos were placed in Phosphate Bufer suftate (PBS) 

with 10% of glycerol, and they were placed individually in 0.25 ml 

strays. An identification was placed in each stray and then we start 

the freezing process, this process consisted in passing from the 

environment temperature to the ethanol bath in the manual freezer at 

a temperature of -6 C to -7 C, after this cristalization induction in 

which they remained for ten minutes, the descent of the temperature 

was made at a 0.5 C/min rate until we reach -35 C, at this 

temperature the strays were submerged In liquid nitrogen, in which 

they were stored until they were used. Only Quality 1 (excellent) 

morulas and blastocyst were frozen. Defrosting was carried out I 

four steps and only the embryos who stayed in the Quality 1 level 

(excellent) were tansfered. Transferece was made through the 

surgical method through an incision in the side ipsilaterally to the 

ovary presenting the corpus luteum.387 embryos were transferred, 

138 were morulas and 249 were blastocyst. Pregnancy diagnosis was 

made 45 days after the transference. The pregnancy rate for morulas 

was of 58.69% and 53.41 for blastocysts there was no stadistical 

significance (Ji square, P 0.05). Total pregnancy rate was 55.29%. 

We conclude that when we transfer embryos maintaining quality 1 

when they are defrosted, there is no difference related with their 

fertility rates when they are transferred. The total pregnancy rate in 

this research, tell us that is profitable the frozen embryo transfer 

technique and that is useful for the bovine genetic improvement. 

928 Effect of Two Sperm Preservation Methods on Conception 

Rate after Fixed-time Artificial Insemination 

A. Crespilho 1 , F. Papa 1 , A. Martins Jr 1 , R. Calegari 1 , 

J. Dell’ Aqua Jr 1 , C. Silvav 

1 Sao Paulo State University, Animal Reproduction and Veterinary 

Radiology, Botucatu-SP, Brazil 

2 Sao Paulo State University - UNESP, Clinical, Surgery and Animal 


Viability of frozen bovine semen is of importance to obtain 

acceptable conception rates (CR). Although much progress has been 

made, several studies concerning sperm fertilizing ability are under 

intensive investigations since there is still considerable variation in 

success rates. This study was conducted to compare the effect of two 

preservation methods on CR after fixed-time artificial insemination 

(FTAI). For this purpose, ejaculate of 1 Nellore (Bos taurus indicus) 

bull was collected by eletroejaculator, polled and divided into two 

equal aliquots. The control (CG) and tested (TG) groups consisted of 

sperm either frozen in Botu-Bov ® (Biotech, Brazil) or cooled in 

BotuBov ® (without glycerol) extender to yield 20 x 10 6 or 5 x 10 6 

spermatozoa/0.5 ml straw, respectively, after appropriate dilution. 

Extended sperm was cooled to 5 °C for 3 h using a refrigerator 

(Mini-Tübe, Germany) and then frozen in liquid N 2 vapor for 20 

min, before final freezing through direct immersion in N 2 (CG) or 

refrigerated and kept at 5 °C in a Botutainer ® (Biotech, Brazil) for 48 

h prior to use (TG). Afterwards, semen samples were utilized for 

FTAI of 147 cows following estrus synchronization. At a random 

day of the estrous cycle (Day 0) all the animals received 2 mg 

estradiol benzoate (Estrogin ® , Farmavet, Brazil) and a progesterone 

intravaginal device (Primer ® , containing 1g of progesterone, 

Tecnopec, Brazil). On Day 6 and 8, 12.5 mg PGF 2a (Lutalyse ® , 

Pfizer, Brazil) was administered and progesterone device was 

removed, respectively. Then, cows received another estradiol 

injection (1 mg) on Day 9 and were inseminated 24 h later. 

Pregnancy was diagnosed by ultrasonography 30 days following 

insemination. Comparison of pregnancy rates between CG and TG 

groups was analyzed using Chi-square test with P

Institute of Physiology and Sanocreatology, Chisinau, Moldova 

Spermogram is the wide used laboratory’s method for researching 

the state of bull’s reproductive system. Sperm or ejaculate is the 

mixture of excretion of men’s genitals secretion products (testicle 

and their appendages, prostate glands, seminal vesicles, 

bulbourethral (Cowper’s) glands, urethra) and consists of the liquid 

from seminal vesicles, liquid from prostate and spermatozoon. 

Collected sperm is putted into test-tube for the centrifugation during 

20 min. with 10 000 r/min. Plasma of sperm in quantity of 1 ml is 

putted into the other test-tube and deposited with the help of 

sulfosalicylic acid (6%). The liquid is mixed and is leaved for 1 hour 

in a refrigerator and after is centrifuged during 30min with 3000 

r/min. The sample has to be stored in place with low temperature. 

Acid from this liquid is evaporated with help of rotary evaporator 

with temperature of 40 °C, adding distilled water periodically till 

time when pH will be 2.2. This process has to be done till the sample 

will be dry out. It is possible to analyze the same without preliminary 

drying out. Sperm is found in the sediment of the sample (3-30mg). 

It is influenced with 1 ml of sulfosalicylic acid (6%), after is 

centrifuged during 30min with 3000 r/min. Acid from this liquid is 

evaporated with help of rotary evaporator with temperature of 40 ºC, 

adding distilled water periodically till time when pH will be 2.2. 

Sediment dissolves in starts lithium buffer (1ml, pH 2.2). Sample is 

stored in refrigerator. The analyze is carried out in standard regime 

of free amino acid’s determination using lithium buffer (pH 2,90; 

2,95; 3,20; 3,80 and 5,00) with the speed of flow 12,0 ml/hour. The 

obtained data were statistically analyzed using Students t-criteria 

method (Mercurieva, 1976). It was revealed that sum of bull’s free 

amino acids in sperm liquid was 1835.4±579.77 micromole/100mg 

but in sperm it was 35.8±17.53micromole/100mg. Analyzing 

obtained data we can make a conclusion that it is corresponding to 

the existing standards and that free amino acids contains in sperm 

liquid more than in sperm. 

931 Definition of Quality Sperm of the Bulls Frozen with High 

Effective Environments at Use of Different Technological 

Modes 

V. Buzan, Gh. Boronciuc, V. Buzan, I. Mereuta 

Institute of Physiology and Sanocreatology, Chisinau, Moldova 

Efficiency of freezing biological objects is defined by components 

of synthetic environments and modes of freezing. These two 

directions are important and require studying at updating or 

improvements of technologies of long storage of sperm of 

agricultural animals. In result the lead experiences it has been proved 

that one of the best-investigated environments is medium lactosesorbite-glycerin-yolk. 

One of the important factors at freezing sperm 

is the stage of cooling. Therefore, in one special experience 

influence of the period of cooling and time of filling of sperm has 

been investigated. For research sperm of bulls of breed Baltata cu 

negru (Holstein) farms containing in conditions was used. Mobility, 

life expectancy and an absolute index have been appreciated with the 

accepted methods. It was used sperm of the bull with mobility of not 

less than 7.0 points and concentration of 0.7 for experiences. With 

medium was 1:5. At crioconservations sperms of the bull in 

polymeric LSTGG environment was used. All experiences repeated 

not less than 5 times. For performance of total problems, experiences 

were carried out under below specified scheme. In experience, 

influence of duration of cooling of sperm is investigated at 2-40. 

Besides it, time of cooling of sperm in a refrigerator before filling 

and after that is investigated. Performance of the given circuit 

promotes definition of optimum time of freezing of sperm of the bull 

and at what height from a surface of liquid nitrogen this 

technological operation demands for realization. Proceeding from 

experiences at use of LSTGG environment for freezing sperm of 

bulls in polymeric the following circuit is offered: 1. Filling 

polymeric sperm at once after it. 2. Cooling a seed material during 3 

hours. Such simplified mode allows receiving quality of the frozen 

sperm, which corresponds to technology requirements. Other series 

of researches has been directed for research of the new environment 

for freezing sperm of bulls. As a result of the lead researches, it is 

possible to draw conclusions: 1. Cooling sperm of bulls is necessary 

to carry out during 3 hours at temperature 2-4. 2. It is possible to 

dilute sperms, to put it in polymeric without its adaptation to new 

conditions. 


932 Effect of Lineage of Oocyte Donors on the Efficiency of OPU 

in Nellore Cows 

A. Martins Jr 1 , R. Calegari 1 , C. Silva 1 , A. Crespilho 2 

1 Sao Paulo State University - UNESP, Clinic, Surgery and Animal 


2 Sao Paulo State University - UNESP, Animal Reproduction and 

Veterinary Radiology, Botucatu-SP, Brazil 

It is well-known that oocyte donors can influence the number of 

recovered oocytes by ultrasound-guided follicular aspiration. 

Thereby, this study was designed to assess the effect of lineage of 

donors on the number of cumulus oocyte-complexes (COCs) 

harvested by ovum pick up (OPU). Fifty-six Nellore (Bos indicus) 

cows derived from two distinct lineages of bulls, named Karvadi (K) 

and Taj Mahal (T) were used. The animals were assigned to four 

groups, according to the maternal (m) or paternal (p) lineage, as 

follows: Kp-Km (n=15), Kp-Tm (n=11), Tp-Km (n=21), and Tp-Tm 

(n= 9). The experiment was conducted over two consecutive years. 

All visible follicles were punctured using a real time B-mode 

scanner (Aloka SSD-500, Japan) and a transvaginal convex 

transducer (5 MHz) to which a needle guide and a suction pump 

were connected. Retrieved COCs were evaluated and classified into 

seven categories (I-VII), graded from the best (I) to the worst (VII; 

degenerated) quality, based on the cumulus cells investment and 

cytoplasmic appearance. The data were transformed into log (x+1) 

before statistical analysis (ANOVA and Tukey’s test) with P

and in group B it is 85.39 days. Furthermore, the mean calving to 

first service interval in group A is 75.9 days and in group B it is 

111.7 days, the mean of days open in group A is 120 days and in 

group B it is 154 days, and the mean calving interval is 401 and 420 

days in group A and B, respectively. Therefore it is concluded that 

the early recognition and treatment of ovarian cysts after calving in 

dairy cows is one of the most important factors influencing the cost 

benefits of a dairy unit. 

Key words: ovarian cyst, cost benefit, dairy cow 

934 Effect of Mineral Supplementation Minerals Level in Seminal 

Plasma of Buffaloes Raised in Amazonia, Brazil 

R. Viana 1 , P. Baruselli 2 , E. Cardoso 3 , C. Araujo 1 , B. Monteiro 1 , 

W. Vale 1 

1 

Universidade Federal Rural da Amazônia, Instituto da Saúde e 

Produçao Animal, Belém, Brazil 

2 

Universidade de Sao Paulo, Faculdade de Medicina Veterinária e 

Zootecnia, Sao paulo, Brazil 

3 

Universidade Federal Fluminense, Faculdade de Veterinária, 

Niterói, Brazil 

The micro-minerals are essential for metabolic reactions of the 

reproductive hormonal functions, for spermatogenesis and the 

process of spermatic maturation. Therefore, it was objectified in this 

study, to determine the nutrition influence in the mineral 

concentrations seminal plasma of buffaloes. Two experiments (A 

and B) had been lead in two farms located in the Eastern Amazonia, 

Brazil. Animals had been kept in pastures of Brachiaria brizantha cv 

Marandu in rotational grazing (GA1-27 buffalos that had consumed 

a mineral mixture for bovines, and GA2-27 buffalos that had 

consumed a mineral supplement for buffaloes), and in continuous 

grazing (GB1-12 buffalos, that had consumed a organic mineral 

mixture for bovines, and GB2-11 buffalos that had consumed a 

mineral supplement for buffalos). After 14 months of mineral 

supplementation, with 22 months of age, had been done 8 semen 

collects, one per week, using electro ejaculator. The semen was 

centrifuged for 3,000 rpm during 20 minutes and the supernatant was 

aliquot and congealed at -20°C. The determination of the P was done 

through the technique of colorimetric and the levels of Ca, Mg, Cu, 

Co, Fe, Zn, Mn done by flame atomic absorption spectrophotometry. 

Exactly presenting equal mineral levels in the grass of both 

the groups, group GA1 that consumed a bigger amount of mineral 

mixture that contained greatest levels of Mn, presented in the 

seminal plasma a smaller concentration of Mn (0,05±0,04 µg/dl) in 

relation to group GA2 (0,09±0,07 µg/dl). In experiment B, was 

verified a smaller availability of Co and Mn, and greatest levels of P 

in the mineral mixture consumed by group GB1 (Mn=0,15±0,20 

g/dl; Co=0,12±0,06 µg/dl and P=4,08±3,12 g/dl) that they had 

presented greatest levels of Mn in the seminal plasma, while in group 

GB2 (Mn=0,05±0,13 µg/dl; Co=0,16±0,06 µµg/dl and 

P=4,70±4,87 g/dl) demonstrated to greatest levels of Co and P. Both 

the groups presented equal consumptions of mineral mixture, and the 

consumed grass had equal mineral levels. In the buffalos raised in 

rotational grazing, it can be verified that the mineral mixture 

influenced in the manganese concentrations of the seminal plasma. 

And in the animals raised in continuous grazing, this influence 

occurred in the levels of phosphorus, manganese and cobalt. 

Key words: buffaloes, reproduction, minerals, semen 

935 Effects of Seminal Plasma Removal on Spermatozoal Motion 

and in vitro Embryo Production 

J. Tanajura Neto, A. Martins Jr, R. Calegari, C. Silva, A. Crespilho 

Sao Paulo State University - UNESP, Clinical, Surgery and Animal 


This study was designed to verify the influence of seminal plasma 

removal on sperm parameters and in vitro embryo production. 

Twenty ejaculates were collected by an artificial vagina from four 

Nellore bulls. Five sperm samples from each of the bull were 

evaluated and used if motility and morphology abnormalities were 

>60% and

2 

University College Dublin, Animal Health and Welfare, Dublin, 


A total of 46 Irish Holstein-Friesian spring-calving cows were 

blocked [lactation number (1 to 8), body condition, body weight, 

calving date, predicted milk yield] and allocated to two management 

environments [cubicle housing with a total mixed ration pre- and 

postpartum (HOUSED) and rotational pasture with grass silage preand 

grass and concentrate postpartum (GRASS)]. The occurrence of 

reproductive disorders [dystocia, retained foetal membranes and 

puerperal metritis, 0-10 days in milk (DIM) and endometritis 

(MetricheckTM) 35-49 DIM], the commencement of luteal activity 

(CLA) (from thrice weekly milk sampling for progesterone analysis, 

EIA) and reproductive performance following a seasonal breeding 

period (98 days), were recorded. Proportions were analysed using 

Fisher’s exact or chi-square while the collated records for 

continuous variables were analysed using Wilcoxon’s or T-test in 

SAS, as appropriate. The raw average milk production for the 

treatments was of 6,186 kg vs. 7,299 kg for GRASS and HOUSED 

cows, respectively. Cows managed in the GRASS environment had a 

lower incidence of dystocia (4.4 vs 17.4%, P=0.08), puerperal 

metritis (17.4 vs 47.8%, P=0.06), endometritis (9.1 vs 31.8%, 

P=0.07) and an earlier CLA (median, and interquintile range: 23, 20- 

31 vs 34, 28-50 days, P=0.09). Overall, reproductive performance 

was suboptimal and did not differ significantly between groups. 

However, it is posited that the lower incidence of uterine disorders 

and earlier CLA of the GRASS cows may have contributed to their 

numerically higher submission rate (82.6 vs 60.9%, P=0.19), shorter 

calving to service (70 vs 75.7 days, P=0.38) and calving to 

conception (102 vs 107.1 days, P=0.29) intervals and higher final 

pregnancy rate (73.9 vs 60.9%, P=0.53). Though this is a 

biologically plausible explanation, limited experimental units may 

have reduced the probability of detecting these numerical 

differences as statistically significant. It is speculated that treatmentinduced 

differences in body condition loss pre- and postpartum and 

differential peripartum immune suppression may have contributed to 

these findings. In conclusion, cows managed in a GRASS 

environment appeared to have an earlier onset of uterine involution 

and luteal activity and this appeared to be associated with better 

reproductive performance however; the limited scale of this study 

precludes robust inference. 

Key words: uterine infection, reproductive performance, pasture vs 

TMR systems 

938 Apoptosis, Placental Maturation and Placental Retention in 

Bos Taurus 

E. Martins 1 , A. Vasconcelos 1 , A. Marques Jr 2 , J. Ferreira Neto 2 , 

J. Nunes 2 , V. Martins 1 , K. Meça 1 

1 

Instituto de Ciencias Biológicas UFMG, General Pathology, Belo 


2 

Escola de Veterinária UFMG, Departamento de Clinica e Cirurgia 

Veterinaria, Belo Horizonte, Brazil 

Apoptosis is an important event for the homeostasis and placental 

maturation. A decreased cellularity in placentome close to 

parturition suggests that placental release is somehow associated 

with that. Here, the role of apoptosis in placental maturation was 

evaluated in 42 Holstein puerperal cows, distributed in 3 treatments: 

parturition to term (Group I, n= 22); parturition to term and retained 

placenta (Group II, n=10) and dexamethasone-induced parturition 5 

days before the predicted time (Group III, n=10). Placentomes were 

collected and processed. Slides stained by Shorr and MGP were 

submitted to morphometry to count caruncular epithelial, 

trophoblast binucleated and apoptotic cells. Apoptosis was also 

approached by the TUNEL, ELISA (to detect free histones), DNA 

electrophoresis (to confirm the internucleosomal fragmentation) and 

transmission electron microscopy. Caruncular cells along 100 µm of 

epithelium did not differ between groups I (3,59±0,59) and II 

(4,28±0,71) (p>0.10). However, group I presented a number of 

caruncular cells smaller than group III (4,75 ± 1,56) (p0.01) than 

in groups II (4,84±10,84) and III (5,02±11,18). ELISA results were 

similar to the morphometric ones, except that groups III and I were 

not different (p

940 The Effect of Linseed Based Inert Fat Supplementation on 

Reproductive and Metabolic Performance in Holstein-Friesian 

Cows 

M. Keresztes 1 , G. Sassi 2 , V. Faigl 1 , M. Kulcsár 1 , H. Fébel 3 , 

M. Mézes 4 , Gy. Gábor 3 , O. Szenci 1,2 , Gy. Huszenicza 1 

1 Szent István University Faculty of Veterinary Science, Reproduction, 


2 Szent István University Faculty of Veterinary Science, Large Animal 

Clinic, Budapest, Hungary 

3 Research Institute for Animal Breeding and Nutrition, Herceghalom, 

Hungary 

4 Szent István University Faculty of Agricultural and Environmental 

Sciences, Department of Animal Nutrition, Gödöll[, Hungary 

Feeding rumen-protected fatty acids to ruminants - particularly if it 

is rich in polyunsaturated fatty acids (PUFA) - can influence 

reproductive activity through a variety of mechanisms. Both n-6 and 

n-3 PUFAs can stimulate the ovarian follicular growth, the luteal 

function and the steroid metabolism of the animals. The objective of 

this work was to determine the effect of a linseed based inert fat 

supplementation rich in linoleic (C18:2, n-6) and linolenic acid 

(C18:3, n-3) on metabolites and hormones in plasma, as well as on 

resumption of ovarian activity during early lactation. Forty-nine 

multiparous (2,73 ± 0,45 BCS) Holstein-Friesian cows were 

assigned in Control (n=25) and fat-supplemented (FS; n=24) groups. 

Beginning 1 day post partum rumen protected fat based on linseed 

oil was added to the TMR diet of the Treated group, throughout 90 

days of the lactation. Metabolizable protein and net energy for 

lactation concentrations were similar in the 2 diets. Blood samples 

were taken on the 1, 2, 5, 10-11 postpartum weeks for quantification 

of ‚OH-butyrate (BHB), non esterified fatty acids (NEFA), urea, 

insulin, insulin-like growth hormone factor I (IGF-I), 3’,3,5triiodtironine 

(T3) and thyroxine (T4) levels. Milk samples were 

taken three times a week from the parturition until the 7 week for 

progesterone (P4) concentration. Average daily milk production in 

the first 180 pp day was higher in the treated group (30.96± 5 kg/d 

vs. 26.66±4.6 kg/d; P

(I.M.) 1 mg of estradiol cipionate to 100 cows for each group 20 

days after I.A. At the time of injection was taken a blood sample in 

order to measure P4 levels. Fertility was calculated in all the cows 

that were inseminated. In all cases, the pregnancy diagnosis was 

carried out for rectal palpation at 40 days after AI. We concludes 

that the injection of 1 mg of estradiol cipionate at 20 days after the 

AI, is effective to diminish the proportion of empty cows to the 

diagnosis during the summer, not affecting the gestation in course 

and that the fertility to the first service of the induced estrus is 

higher in winter. 

Key words: dairy cow, reproduction, infertility 

944 Correlation between the Minerals Levels of Seminal Plasma 

and Semen Quality of Buffaloes Raized in Amazonia, Brazil 

R. Viana 1 , P. Baruselli 2 , E. Cardoso 3 , C. Araujo 1 , B. Monteiro 1 , 

W. Vale 1 

1 

Universidade Federal Rural da Amazônia, Instituto da Saúde e 

Produçao Animal, Belém, Brazil 

2 

Universidade de Sao Paulo, Faculdade de Medicina Veterinária e 

Zootecnia, Sao Paulo, Brazil 

3 

Universidade Federal Fluminense, Faculdade de Veterinária, 

Niterói, Brazil 

This study had as aim to verify the mineral correlations in the 

seminal plasma and the characteristics of the semen of buffaloes. 

The animals were kept in pastures of Brachiaria brizantha cv 

Marandu in rotational grazing (n=54) and in continuous grazing 

(n=23). Eight semen collections were performed (one per week, 

using electro ejaculator) after 14 months of mineral 

supplementation, at 22 months of age. After made morphologic and 

physicist-chemistries evaluations of the spermatozoa, semen was 

centrifuged for 3,000 rpm during 20 minutes and in the supernatant, 

were determined P through colorimetry and the levels of Ca, Mg, Cu, 

Co, Fe, Zn, Mn by flame atomic absorption spectrophotometry. The 

averages of the evaluation of minerals in the seminal plasma had 

been analyzed by the test of Kruskal-Wallis. Also correlations of 

Pearson had been carried between the minerals concentration in the 

seminal plasma and the characteristics of the semen. In the animals 

created in rotational grazing significant correlations of the Cu 

(r=0,24) and Mn (r=-0,28) with the scrotal circumference had been 

observed; of the Co (r=-0,23) with spermatic concentration; of the Fe 

with the gradual motilities (r=-0,24) and with the mass motility (r=- 

0,31); of the Ca with the total of spermatic defects of head (r=0,26); 

of the Mg with the total of spermatic defects of main part (r=-0,19) 

and of tail (r = 0-0,18); and of the P with the gradual motilities 

(r=0.21), with the mass motility (r=0,45) and with the total of 

spermatic defects of head (r=0,26). In the animals created in 

continuous grazing significant correlations of the P with the gradual 

motilities had been observed (r=0,52), with the spermatic vigor 

(r=0,43), with the mass motility (r=0,53), with the spermatic 

concentration (r=0,44), with pH (r=-0,38), with the total of defects of 

intermediate part (r=0,51), with the total of spermatic defects of 

main part (r=0,45) and of tail (r=0,40); and of the Zn with pH (r=- 

0,40). Although some minerals have presented significant 

correlations with some characteristics of the semen of the buffaloes 

created in rotational and continuous grazing, these had been low. 

Therefore, the qualitative and quantitative variation in the physical 

and morphologic concentrations of the semen of buffalos was not 

observed, in function of the mineral concentrations presents in the 

seminal plasma. 

Key words: buffaloes, minerals, seminal plasma, semen 

945 Antibiotic Sensitivity (MIC) of Bacteria Isolated from 

Puerperal Metritis of Dairy Cows 

J. Földi 1 , J. Szabó 2 , B. Huyghe 3 , C. Sa 3 , A. Pécsi 4 , M. Kulcsár 3 , 

P. Cox 3 , J. Swinkels 6 , G. Huszenicza 5 

1 Intervet Hungaria Kft, Budapest, Hungary 

2 Faculty of Medical and Health Science - University Debrecen, 

Clinical Bacteriology, Debrecen, Hungary 

3 Intervet Schering Plough, Angers, France 

4 Faculty of Agricultural Sciences - University Debrecen, Animal 

Health and Physiology, Debrecen, Hungary 

5 Faculty of Veterinary Science, Szent István University, Department 

of Obstetrics, Reproduction and Herd Health, Budapest, Hungary 

6 Intervet Schering Plough, Int. Marketing, Boxmeer, Namibia 


A wide variety of bacteria can be isolated from the uterus of the cows 

with puerperal metritis (PM). However, only some of them i.e. 

Arcanobacterium pyogenes, Escherichia coli and Gram-negative 

anaerobes play significant role in the pathogenesis of the disease. 

Uterine swab samples (n=41) were collected in 5 large scale dairies in 

Hungary in 2004, from cows with PM before treatment to screen the 

bacterial flora and determine the minimal inhibitory concentration 

(MIC) of oxytetracyclin (OTC), amoxicillin, cefapirin and cefquinome 

to the primary uterine pathogens. Isolation and identification of the 

bacteria were performed according to the conventional bacteriological 

techniques. The strains were identified up to species levels using 

appropriate biochemical tests and Bio-Merieux ATB automatic 

identifying system. For the determination of MIC 90 µl amounts from 

broth containing two-fold concentration increments of antimicrobial 

agents were added to 96-well microdilution trays. Each well was 

inoculated with 10 µl bacterial suspension (the inoculum was 1.5 x 10 5 

CFU/ ml). The MIC was read as the lowest concentration of the 

antibiotic at which no growth was recorded.The main isolated 

pathogens were as follows: A. pyogenes (n=31), E. coli (n=32), nonpigmented 

Prevotella strains (P. bivia n=24), black pigmented 

Prevotella spp. (n=9), Bacteroides spp. (n=9) Porphyromonas spp. 

(n=2), F. necrophorum (n=4). High MIC (> 1.0 g/µml) was measured 

against OTC at more than half of E. coli and almost all Gram-negative 

anaerobes (except for the few strains of F. necrophorum). Also about 

25% of A. pyogenes strains were resistant to OTC. All E. coli and all 

Bacteroides spp. had of high MIC to amoxicillin (MIC ≥ 1.0 g/m µl and 

MIC ≥ 8.0 µg/ml, respectively). Other Gram-negative anaerobes 

showed variable MIC, while majority of A. pyogenes strains possessed 

relatively low MIC (≤ 1.0 µg/ml) to amoxicillin. The most favourable in 

vitro sensitivity pattern of all isolated primary uterine pathogens (MIC 

≤ 1.0 µg/ml) was measured for cefquinome, a recent generation 

cephalosporin derivate. A. pyogenes and Gram-negative anaerobes 

showed 100% sensitivity against cefapirin, however, 75% of the E. 

coli strains had MIC ≥ 4.0 µg/ml and the remaining 25% were even 

around 1.0 µg/ml. We conclude, that increasing MIC of OTC and 

amoxicillin against the main uterine pathogens is remarkable, as 

compared to our previous findings. 

946 Effect of Subclinical Metabolic Disorders on the Incidence of 

Stillbirth on a Large Scale Dairy Farm in Hungary 

A. Tirián 1 , E. Brydl 1 , O. Szenci 1 , L. Könyvesv, V. Jurkovich 1 , 

L. Tegzes 1 , K. Ari 1 , Cs. Á. Bajcsy 1 , J. Tibold 2 



Hungary 

2 Agroprodukt Co., Pápa, Hungary 

Objectives of the study: was to investigate the possible effect of 

subclinical metabolic disorders on the incidence of stillbirth in dairy 

cattle on a large scale dairy farm in Hungary. 

Materials and Methods: Altogether 160 multiparous dairy cows were 

sampled three times during the past two years in the periparturient 

period (3 weeks before expected calving, < 1 h after calving, and 25 to 

35 days after calving. Pregnant heifers (n=20) were investigated from 3 

months of gestation until calving once a month, then < 1h after calving 

and 25 to 35 days after calving. Blood parameters checked were as 

follows: Glucose-, Beta-Hidroxi-Butirate-, Free Fatty Acid-, Urea-, 

Total Protein-, Albumin-concentration, AST-activity, Carotene-level, 

Ca-, P- and Mg-concentration. Urine parameters were as follows: pH, 

Net Acid Base Excretion. 

Results: Comparing the data of the cows with normal calving (Group 

1, n=189) and stillbirth (Group 2, n=10) only FFA-concentration prior 

to calving showed a significant difference (0,110, 0,143, p

J. Hajurka, I. Valocky 

University of Veterianary Medicine, Kosice, Slovakia 

During the past fifty years pregnancy rates of lactating dairy cows have 

decreased progressively. First ovulation or luteinization of a dominant 

follicle can be induced with exogenous GnRH, and PGF 2a will shorten 

any luteal phases; thus more ovulations can be introduced before first 

insemination. The objective of this study was to determine the effects 

of these treatments on subsequent reproductive performance in dairy 

cows. After day 10 cows were treated with a synthetic GnRH 

analoque (gonadorelinum, 200 µg; i.m.) if a large follicle (> 10mm) 

was present on the ovaries, or with synthetic PGF 2a (cloprostenolum 

500 µg; i.m.) if a CL was present. The animals were divided into two 

groups with normal (n = 81) or gynaecologically abnormal puerperium 

(retained placenta and/or uterine inflammation; n = 61). Cows in the 

experimental group (EG) tended to have fewer days open (85.5 ± 6.4 

days) than the control group (CG; 113.0 ± 15.8 days; P = 0.0524) in a 

normal puerperium with a similar number of inseminations per 

conception (2.0 ± 0.1 vs. 1.9 ± 0.2; P = 0.1252). Treatment in abnormal 

puerperium had no effect on the days open (103.0 ± 8.7 vs. 118.0 ± 15.9 

days in EG and CG resp; P = 0.3936) nor on the number of 

insemination per conception (2.3 ± 0.2 vs. 2.4 ± 0.3 in EG and CG ; p 

= 0.6711). Supported by the Ministry of Education of the Slovak 

Republic, project AV 4/0009/07 

Key words: dairy cows, reproductive performance 

948 High Hydrostatic Pressure Treatment of Fresh Bull Semen 

Increases the Proportion of Cells Surviving the Freezingthawing 

Procedure in a Hungarian Holstein Friesian Bull 

Population 

G. Kútvölgyi 1 , A. Horváth 2 , I. Oláh 3 , P. Pálinkás 3 , L. Merész 3 , 

M. Molnár 1 , CS. Pribenszky 1 , O. Szenci 2 

1 Cryo-Innovation Ltd., Budapest, Hungary 

2 Szent István University Faculty of Veterinary Science, Large Animal 

Clinic, Üll[, Hungary 

3 Bos-Genetic Ltd., Martonvásár, Hungary 

216 Prevalence of Claw Disorders and the Need for Regular 

Examination of Claws in Dairy Cows from Smallholder Units 

of Peri-urban Nairobi, Kenya 

J. Nguhiu-Mwangi 1 , P. Mbithi 1 , J. Wabacha 1 , P. Mbuthia 2 

1 

University of Nairobi, Department of Clinical Studies, Nairobi, 


2 

University of Nairobi, Veterinary Pathology, Nairobi, Kenya, Kenya 

Coast Republic 

A cross-sectional study was conducted in 32 zero-grazing 

smallholder dairy units in the peri-urban regions of Nairobi, Kenya. 

The objective of the study was to establish types and prevalence of 

claw lesions affecting dairy cows under varied zero-grazing 

conditions. The farms were purposively selected and systematic 

sampling technique employed to select cows in each farm. A total of 

300 cows were selected and examined. They were evaluated for 

lameness with standard locomotion scoring system. All hind limb 

claws of the 300 cows were visually examined for lesions after 

thorough washing and trimming. Trimming was useful as 

a diagnostic and corrective procedure of claw disorders. Of the cows 

examined, 88% had claw lesions and among these, 69% of them had 

subclinical lesions. Prevalence rate of each claw disorder was 

calculated separately out of 300. Some cows had more than one claw 

disorder and some claws had more than one disorder. Total 

prevalence of laminitis was 70.3% with subclinical laminitis at 

49.3% and chronic laminitis at 21%. Other claw lesions probably 

related to laminitis had prevalence rates as follows: sole bruising 

(45%), heel erosion (27.3%), white line separation (18%) and double 

or underrun soles (17%). Sole ulcers had low prevalence of 2.7%. 

Claw deformities occurred concurrently with chronic laminitis and 

Lameness 

Previously, we reported that high hydrostatic pressure (HHP) 

significantly improves post-thaw survival of frozen IVP bovine 

blastocysts, presumably from the induction of shock proteins. We 

also have reported increased post-thaw survival of HHP-treated boar 

and bull semen. We now report further data on the effect of HHP 

treatment on motility of frozen thawed bull semen. HHP treatments 

were executed by a computer-controlled pressurizing device. The 

semen of 20 of the production bulls of Bos Genetic, Hungary has 

been produced with or without (HHP) pre-treatment. After semen 

collection the ejaculate was diluted 1:1 with Bioxcell extender at 34 

°C. Diluted semen was split individually, one half was filled into 

maxistraws (treatment groups) and were closed by plastic balls. The 

other half – control group – was further diluted to reach the final 

concentration of 8x10 7 /ml. Control semen was filled to 0.25ml 

straws at room temperature, and were placed to a cooling chamber at 

5 °C for at least 4 hours. After cold acclimatization semen was 

frozen using standard program and was placed into LN 2. The 

treatment groups were left to cool down to room temperature for 30 

min and were placed to the chamber of the pressurizing device. 30 

MPa pressure was applied for 90 min, at room temperature. After 

treatment semen was further diluted at room temperature to reach 

final concentration of 8x10 7 /mL, and was placed to 5 °C for at least 

2 hours before being frozen. Straws were thawed in a 38 °C water 

bath for 30 s. Progressive motility was assessed by the CASA 

system. Experiments were replicated twice or three times for each 

bull. Paired t-test was used to analyze data. HHP treatment 

significantly increased the post-thaw total and progressive motility 

of the bull population (total motility: 54.8% vs. 62.3%; progressive 

motility: 42.2% vs. 47.6%; control vs. treated, respectively). At 6 of 

the 20 bulls (30%) there was no difference in the motility parameters 

between the groups. At 10 bulls (50%) the HHP treatment has 

increased the number of motile cells with an additional 5-15 per cent, 

while at 4 bulls (20%) the treatment related increase in the motile 

cells was between 15 and 25 %. Field trials are being made to 

confirm results in vivo. 

Trial was supported by Kozma grant OMFB-00504/2007 

had prevalence of 30.3%. They included excessively overgrown, 

flattened, concave and grooved claws. Splayed and corkscrew claws 

had prevalence of 8.7% and 4.7% respectively. Other lesions 

involving the claws included infections, trauma, growths and foreign 

bodies which together had prevalence of 6.4%. Cows with chronic 

laminitis and concurrent severe claw deformities were significantly 

associated with higher locomotion scores (Khi 2 = 74.05, P< 0.05) 

and were severely lame. In all, 80% of the cows were raised on 

varied concrete floors such as over-smooth or pot-holed and some 

with loose stones and most of these had slurry that was left on the 

floor for between 24 and 72 hours. The other 20% of the cows were 

on wet and often muddy earthen floors. In conclusion, the cows in 

the smallholder units in the peri-urban Nairobi are raised under 

conditions that promote development of laminitic claw disorders and 

in which lack of regular trimming promotes claw deformities. It is 

therefore pertinent that regular claw check-up and trimming be 

practiced in these farms to promote claw health and animal welfare. 

217 The use of Infrared Thermography (IRT) in Detecting 

Lameness in Dairy Cows 

C. Gelfert 1 , A. Onmaz 2 , G. Atalan 3 , V. Günes 2 , A. Düzler 4 , 

I. Nur 4 , M. Kibar 3 

1 

University of Veterinary Medicine, Clinic for Ruminants, Vienna, 

Austria 

2 

Erciyes Üniversitesi Veteriner Fakültesi, Iç Hastaliklari Anabilim 

Dali, Kayseri, Turkey 

3 

Erciyes Üniversitesi Veteriner Fakültesi, Department of Surgery, 


4 

Erciyes Üniversitesi Veteriner Fakültesi, Department of Anatomy, 

Kayseri, Uganda 

Lameness 217

Lameness results in major economic losses on dairy farms. Infrared 

thermography (IRT) is a non-invasive technique to detect thermal 

radiation from the surface of any object. The objective of this study 

was to evaluate the capability of IRT to detect relationship to visual 

abnormalities of the hoof of dairy cows. In a dairy farm with a 

sudden increase of acute lameness, all cows underwent clinical and 

orthopaedic examination. The temperature of the skin of the claws 

was estimated by manual palpation. Then infrared images were taken 

from all hoofs of 50 lame cows. To compare the temperature 

measured IRT images were taken from 50 non-lame cows from 

another farm without any acute occurrence of lameness and 50 nonlame 

calves. The measured IRT temperature at the single hoof was 

compared statistically using simple ANOVA. The mean maximum 

IRT-temperature of lame claws was 33.38±1.21 °C and 32.40±1.36 

°C of the healthy claws of the same animal (p

findings radiography revealed in three animals no, in one animal a 

partial, and in four cases a complete osseous ankylosis of the DIJ. 

Conclusion: According to the histological and radiological results 

of this study formation of complete osseous ankylosis after resection 

of the DIJ in dairy cows appears to take considerably longer than two 

years. Thus, it remains questionable if the application of any 

measures for only about three month will contribute substantially to 

the development of ankylosis. However a conclusion about the 

clinical mobility of the DIJ is not possible, because even joints with 

a fibrous connection can be clinically fixed. 

259 Long-term Observations in Dairy Farms with Lameness 

Problems 

K. Muller 1 , T. Eilers 1 , R. Pijl 2 

1 Clinic for Ruminants and Pigs, Faculty of Veterinary Medicine, 

Freie Universitaet, Berlin, Germany 

2 Functional Claw Trimming, Jever, Germany 

Objectives of Study: Lameness forms a major problem on dairy farms 

in Germany causing substantial economical losses and impairment of 

animal well-being. In the present study the effects of the 

implementation of functional claw trimming were examined on four 

dairy farms by long-term observations including monitoring of distinct 

cow signals in two-week intervals within a period of one year. 

Materials and Methods: Four dairy farms, keeping between 60 and 

120 dairy cows and sharing common characteristics with respect to 

management, housing conditions and mean annual milk yield, were 

included in this study. The housing conditions were evaluated by a 

standard protocol, and claw disorders were documented at functional 

claw trimming performed three times per year. In addition, distinct 

cow signals as lameness, body condition, stall standing index, 

rumination index, rumen filling and milk production data were 

documented in intervals. 

Results: None of the farms achieved the recommended number of 

points with respect to housing conditions (109±137 points/178 points 

possible). The farms were deficient in walking space, cubicle comfort 

and floor condition. Following functional claw trimming, the severity 

of lameness decreased significantly. The prevalence of infectious and 

non-infectious claw diseases, however, did not change. 

Conclusions: On all farms similar parameters of cow comfort were 

shown to be deficient. The latter deficiencies have been reported to 

affect claw health before. Functional claw trimming is a useful tool to 

improve lameness in herds, but seems to have hardly any effect on claw 

disease prevalence. On basis of these results we assume that only a 

change in housing conditions, especially in cubicle comfort, will have 

a sustainable effect on claw health. 

260 Associations between Milk (Daily Milk Yield, Percentage of 

Milk Protein, Fat and Fat-protein Ratio) and Occurrence of 

Claw Disorders in Holstein Cows 

O. Penkava, P. Fleischer, S. Slosárková, R. Dvorák 

University of Veterinary and Pharmaceutical Sciences Brno, Clinic of 

Ruminant Diseases, Brno, Czech Republic 

Using data from a separate study we evaluated associations among 

milk yield, milk components and occurrence of claw lesions. A total 

of 65 pluriparous cows from a commercial herd (405 cows, mean 

yield 7.980 kg, fat 3.9%, protein 3.3%) have been studied. They 

were kept in free stall on concrete floor with bedding in boxes and 

fed by TMR. Claw trimming was performed at the start of the study 

in April, at the end in October and continuously in lame cows. At the 

start cows were 93 days post partum (p.p.), on average. Claws were 

examined 5 times at regular intervals. We put into evidence and 

scored ulcers (toe, sole ulcer; white line disease/ a feet: 0-4) + sole 

hemorrhage (a feet: 0-1); horn defects (double sole, vertical fissure, 

horizontal grooves, separation of the side wall/ 0-1) and heel horn 

erosion (0-1). Findings were divided according to days p.p. into 10 

groups (30, 60, 90±15 days, etc.), n=25-36. Findings were coupled 

with data on daily yield, % protein, fat and fat-protein ratio (FP) 

from the nearest monthly performed milk yield test. Associations 

among milk and prevalence of claw disorders were examined using 

data of the nearest and 3 previous tests. We employed Spearman´s 

Rank Correlation Test. Following associations were significant 

(p‹0.05). Ulcers and/or sole hemorrhages at 3 months (M) p.p. were 

positively correlated to % of fat and FP at 2 Mp.p. Likewise, cows 

affected in this way at 4 Mp.p. had higher % of fat and higher FP at 

1 and 2 Mp.p. Similarly, cows with horn defects at 3 Mp.p. had 

higher % of fat and FP 1 and 2 Mp.p. Cows with higher % of fat and 

higher FP (a strongly negative energy balance) at the start of 

lactation may be expected to suffer from higher prevalence of sole 

hemorrhages, ulcers and horn defects during the next M. Cows 

affected by heel horn erosion at 5 resp. 6 Mp.p. had at 2 and 3 resp. 

4 Mp.p. lower FP due to lower % of fat (acidotic load). In cows with 

horn defects at 6 resp. 7 Mp.p. there was lower % of fat, resp. protein 

and higher FP at 5 Mp.p. Cows with horn defects at 8 Mp.p. had 

higher yield and lower % of fat at the respective and previous M. 

They had lower % of protein at 6-8 Mp.p. Cows producing more 

milk low in % of fat and protein suffered more frequently from horn 

defects. Such associations became clear only after 5 M from the 

previous claw trimming, probably due to the rate of horn growth. 

This study was supported by Grant MSM6215712403. 

Key words: claw disorders, horn quality, milk yield, milk composition, 

energy balance 

261 Prevalence and Factors Affecting Lameness in Commercially 

Raised Dairy Cows 

G. Oikonomou 1 , A. Kougioumtzis 1 , A. Tsiaras 2 , G. Arsenos 1 , 

G. Valergakis 1 , G. Banos 1 

1 



2 

Vivartia Avee, Athens, Greece 

Cow lameness is one of the main issues concerning the dairy 

industry today. The objective of this study was to assess prevalence 

and factors affecting lameness, as well as its association with 

involuntary culling.A total of 497 primiparus Holstein cows that 

calved between January 2005 and June 2006 were considered. These 

animals were raised in a large commercial farm in N. Greece and 

housed in 4 free stall barns with concrete flooring. Cow locomotion 

was scored weekly from calving to week 13 of lactation and 

thereafter monthly until the end of lactation, using a 1 to 5 scale. 

Cows were then defined as healthy during the 305 day lactation (all 

locomotion scores = 1) or lame (at least one locomotion score ≥ 2), 

and lameness was treated as a binary trait (0/1). Measurements of 

body condition score, estimated live weight, energy content, and 

blood serum levels of glucose, -hydroxybutyrate and non-esterified 

fatty acids were also obtained for these cows, once two months 

before calving and once again during the first week post calving. 

These measurements, together with factors such as calving season, 

calving year, age at calving and country of origin, were examined for 

their effect on lameness with the use of a mixed linear model. 

Another model was used for the estimation of lameness effect on 

cow reproductive performance; the latter was evaluated with both 

conception and interval traits. 46% of all cows were at least once 

lame and 31.5% were culled during first lactation. 60% of the culled 

cows were lame before culling and lameness was the main reason for 

involuntary culling. Calving season and live weight at first week of 

lactation had a significant impact on the prevalence of lameness 

(P0.05). 

Lameness was also found to have a significant (P

methods; ANOVA (with binary classification of lameness), ANOVA 

with a lameness index (LAMIX) as the variable of interest, and 

ANCOVA (controlling for mean milk production for the first 3 

weeks of lactation and with a binary classification of lameness). 

Data were from one dairy farm located near Ithaca, NY, from the 

calvings that occurred from 1 January 2004 to 1 January 2007. The 

LAMIX was defined as: -1, -2, -3, -4, and -5 = 1, 2, 3, 4, and 5 

months before lameness was detected respectively; 1, 2, 3, 4, and 5 = 

1, 2, 3, 4, and 5 months after diagnosis respectively; and 0 = cows 

that were never lame. With binary lameness classification analyzed 

by repeated measures ANOVA, there was no effect of lameness on 

milk yield. This model was biased because lame cows had higher 

milk yields prior to lameness compared to non-lame cows. When the 

lameness index was used, milk production prior to lameness was 

greater than after lameness. However, point estimates generated for 

lameness index can be inaccurate because of the multicolinearity 

detected between lameness index and week of lactation, and also 

because of the inability of adjusting the LSM for the interaction of 

lameness index and week of lactation. Therefore, we concluded that 

the most appropriate models were the ANCOVA models (both for 

the matched and non-matched retrospective-cohort designs). The 

estimated losses associated with lameness were 314 and 424 kg/cow 

305-d lactation, respectively, for the matched and non-matched 

designs. Furthermore, high milk yield in the beginning of the 

lactation was a risk factor for lameness. 

263 Functional Trimming at the Dry – off Period to Prevent Foot 

Diseases in Confined Dairy Cows in the Tropical Area 

D. Garcia, M. Hahn, D. Pino, I. Vivas, M. Leal, K. Clerc 

Universidad Del Zulia, Medico Quirurgico, Maracaibo, Venezuela 

The aim of the present research was to evaluate the functional 

trimming effect at the dry off period to prevent the hoof diseases in 

confined milking cows. To do that 47 Brown Swiss cows with two or 

more parturition were study at the beginning of the dry off period. 

The cows were divided in two groups in a random way. One group 

received the functional trimming as treatment and the other one 

serve as a control. The cows were monitor to look into the 

appearance of hoof diseases within the first 5 months of the milk 

yield period. The results show that 39% of the hoof had any kind of 

lesions and 61% of the hoof were without lesions, in the same way 

the treated animal group show only 4% of sick animals whereas in 

the control group were 45.4%. A comparison study of both groups 

show a significant statistically differences (p

same protocol. Cattle having a combination of predetermined 

lameness, swelling and lesion scores on one foot for two 

consecutive days were enrolled. Day 0 was the day the cattle met 

the enrollment criteria, were allotted to treatment and administered 

either tulathromycin or saline control according to a generalized 

block design.Treatment success was analyzed using a mixed model 

which included the fixed effect of treatment and the random effects 

of block and residual. Samples were obtained from the lesions prior 

to treatment for anaerobic culture and sensitivity testing. 

Presumptive identification of isolates obtained from the samples 

was based on classical biochemical methods. Final identification of 

the isolates was based on PCR amplification of the 16S ribosomal 

gene. The agar dilution method described by the Clinical and 

Laboratory Standards Institute was employed for determination of 

tulathromycin MICs. Cattle were clinically evaluated on day 7 and 

treatment success was determined based on defined decreases in 

lesion, swelling and lameness scores. The number of animals 

classified as a treatment success for the Nebraska site was 

significantly greater, P

Initially two radiographs were obtained in latero-medial and 

dorsopalmar/dorsoplantar positions. Then the claws were dissected to 

determine the anatomical position of the distal sesamoid 

bone. Morphologic study revealed that, distal sesamoid bone varied 

from elliptical to rectangular shape and had two surfaces and four 

borders. Its articular surface was smaller, concave and divided into two 

unequal parts by a sharp sagittal ridge in which medial part was more 

extensive and deeper. The palmar/ plantar surface was larger, slightly, 

convex and had a blunt sagittal ridge. Distal sesamoid bone together 

with distal and middle phalanges formed the distal interdigital joint 

(coffin joint). Deep digital flexor tendon passed over the palmar/plantar 

surface of the bone and the podatrochlear bursa was located between 

this tendon and distal sesamoid bone. Distal interdigital ligaments had 

2 strong bonds, which were attached proximally to abaxial eminences 

on the proximal ends of the middle phalanges, then crossed the deep 

digital flexor tendon obliquely and reached the interdigital space and 

finally attached to the distal sesamoid of the opposite 

side. Radiographic study showed that the distal sesamoid bone was 

elliptical and its palmar / plantar surface was slightly convex with a 

blunt sagittal ridge. Its distal border lies to the last quarter of distal end 

of middle phalanx and the proximal border lies at middle of 2nd 

phalanges. In the dorsopalmar/dorsoplantar view, the distal sesamoid 

bone was covered width the middle phalanx. This bone in lateral view 

was diagonal and its density was almost similar to the middle phalanx. 

The trabecular patterns of osseous tissue were visible in the lateral 

view. 

Key words: cattle, morphology, radiography, distal sesamoid 

952 Relationship between Lameness, Udder Health and Milk 

Yield in Dairy Cows 

M. Feldmann, K. Maier, M. Hoedemaker 

University of Veterinary Medicine Hannover, Clinic for Cattle, 


It was the aim of this study to investigate the effect of lameness on 

udder health and milk yield in dairy cows. The data were collected 

from 839 cows of a large dairy farm in Saxony during one lactation. 

Locomotion scoring was performed at the time of the regular claw 

trimming sessions. Cases of lameness occurring between those 

sessions were recorded by the claw trimmers. As to milk yield, 

somatic cell count and incidence of mastitis, lame cows and cows 

without lameness were compared regardless of the lameness 

diagnosis. Furthermore, the effect of lameness score (LS; 1 = mild, 2 

= moderate, 3 = severe) was investigated. From a total of 839 cows, 

297 animals were lame at least once during lactation (35.4 %). 305d 

milk yield did not differ between lame and non lame cows. 100-d 

milk yield was lower in lame cows with > 2 lactations than in non 

lame cows with > 2 lactations (3812 ± 617 kg vs. 4041 ± 661 kg; P < 

0.05). However, based on the monthly milk recordings, the milk 

yield close to the time of lameness diagnosis was lower in lame cows 

than in corresponding non lame cows. This effect was observed for 

cows with 1 and 2 lactations during early lactation and for cows with 

> 2 lactations throughout the whole lactation. Furthermore, there 

was no effect of the degree of lameness on cumulative milk yield. 

Mean log somatic cell count (log SCC) throughout lactation and log 

SCC related to the occurrence of lameness did not differ between 

groups. Incidence of clinical mastitis did not differ between lame 

and non lame cows (47.1 % vs. 48.3 %; P > 0.05). The average 

mastitis cases per animal and lactation as well as milk loss due to 

withdrawal time did not differ between lame animals (2.64 cases per 

cow, 20.2 d per cow) and non lame animals (2.67 cases per cow, 19.7 

d per cow). There was no relationship between the degree of 

lameness and the number of mastitis cases per cow. In conclusion, 

our results suggest that lameness only had a minor effect (older cows 

only) on cumulative milk yield, but that the milk yield around the 

time of lameness diagnosis was negatively affected. Furthermore, in 

this study, no relationship between lameness and udder health was 

found. 

Key words: lameness, udder health, milk yield 

953 Performance of a Hoof Care Solution - DeLaval Hoofcare DA 

- used on Different Farms 

X. Goossens 1 , G. Cavalcanti 2 , T. Hemling 2 

1 DeLaval, PP Milk Quality & Animal Health, Gent, Belgium 


2 

DeLaval, PP Milk Quality & Animal Health, Kansas City, United 

States 

Digital dermatitis (DD) is one of the most common causes of 

lameness in dairy cattle throughout the world. Its importance has 

grown exponentially lately, as it has shown its economical losses 

impact due to increased culling rates, high treatment costs, decreased 

milk production and lowered reproductive efficiency. It also 

represents an animal welfare concern for the dairy industry, as this 

contagious disease can cause pain and discomfort. The goal of this 

study was to evaluate the performance of DeLaval Hoofcare DA in 

the control of digital dermatitis on different commercial dairy farms 

in different countries. DeLaval Hoofcare DA is a quaternary 

ammonium-based (quat) product that has similar therapeutic and 

prophylactic properties to copper sulphate and formalin, but without 

the adverse ecological or health hazards of these products. Six trials 

were performed using a split footbath design. On each farm, one of 

the split footbath’s sides contained Hoofcare DA. DeLaval Hoofcare 

DA was compared against CuSO4, formalin, an experimental quatbased 

product, and an experimental non-quat product. The farms 

were located in Belgium, UK, US and Canada. Hooves were scored 

in a 2-4 weeks interval in the milking parlour, during milking. The 

same consistent scoring system was used in all trials, and for each 

farm, the same trained individual performed all hoof evaluations. All 

trials had the same footbathing regiment of once daily after milking, 

with the same product concentration. Nevertheless, the results varied 

greatly among farms. From the range of results, Hoofcare DA 

reduced the number of DD lesions between 7.9% in 12 weeks and 

80% in 6 months, with an average of 36.6% of reduction. All studies 

showed that footbathing with Hoofcare DA improve the overall 

disease condition. Footbathing alone can not be the factor of success 

if other risk factors are not being well managed. These results 

suggest that a control group should be used on footbath studies to 

avoid comparisons of successful cases on one farm with poor results 

on a different farm, due to differences in hoof health management. 

The split footbath design is a good way to perform a trial testing new 

footbath solutions. Keeping digital dermatitis under control should 

be the key objective of footbathing, since the complete elimination 

of the problem, after the disease has affected a herd, is very difficult 

to achieve. 

954 Dispersion and Grade of Sole Hemorrhages in Subclinical 

Laminitis in Cattle 

M. Rasekh 1 , K. Sardari 2 , H. Seifi 2 , A. Raji 3 , M. Mohri 2 , 

H. Moaddab 1 , H. Mohammadi 1 




Department of Clinical Science, Mashhad, Iran 


Department of Basic Veterinary Sciences, Mashhad, Iran 

Laminitis is a diffuse aseptic inflammation of the laminar corium of the 

hoof wall, a process that involves vascular breakdown, hemorrhaging, 

and exudation of the serum from capillary beds. Approximately 62% of 

the total of hoof lesions can be associated with laminitis. Such 

laminitis-related lesions have a greater economic importance than 

infectious diseases. Subclinical laminitis is the most common form of 

laminitis in dairy cows and lesions associated with subclinical laminitis 

are: sole ulcer, double soles, hemorrhages, etc. Inflammation of the 

corium results in hemorrhaging of sole which is clinical sign and sequel 

lesion of subclinical laminitis. In this study, 491 dairy cows of sixteen 

farms near Mashhad, Iran, were examined. The yellow discoloration 

and hemorrhage of the sole were observed lesions associated with 

subclinical laminitis. According to the international standard 

classification of sole, the dispersion of sole hemorrhages was evaluated 

from region 1 to 6. In addition the degree of sole hemorrhages was 

scored with severity and distribution from 0 to 5. Seven hundred two 

among three thousand nine hundred twenty eight digits were affected to 

subclinical laminitis. Data were analyzed by chi-square test using spss 

programme. According to the international standard classification of 

sole, the dispersion of sole hemorrhages were in regions 4,5,6 more 

than 1,2,3. In addition, the most common type of sole hemorrhage was 

grade 4, severe hemorrhage in small area. The severity of hemorrhages 

is related to the intensity and duration of the insult that caused the

vascular disruption. The site of sole hemorrhaging or ulceration is a 

reflection of the severity of the disease process. 

Key words: subclinical laminitis, sole hemorrhage, cattle, Iran 

955 Foot Lameness and Major Contributing Lesions in Lactating 

Cattle and Buffaloes in Ludhiana District of Punjab, India 

S. Randhawa, K. Dua, C. Randhawa, P. Dhaliwal 

Guru Angad Dev Veterinary and Animal Sciences University, 1- 

Department of Clinical Medicine, Ludhiana, India 

An epidemiological study on foot lameness and major contributing 

lesions in lactating cattle and buffaloes on organized and unorganized 

farms was undertaken. Organized farms were categorized as farms with 

proper animal husbandry practices and herd size above 10 animals and 

unorganized as farms in which recommended animal husbandry 

practices are not properly followed and herd size

number of positive faecal samples divided by the number of samples 

taken. Herds with low (LM) and high mortality (HM) were compared 

using Wilcoxon rank-sum test and Chi-square test. The mortality 

risk was 0-2% (median 1%) in LM herds and 6-24% (median 9%) in 

HM herds. The proportion of dead calves that died at young age were 

sigificantly higher in HM herds (p=0.004) than in LM herds. The 

pathogen score was significantly higher in HM herds (p=0.04), 

indicating that infections with multiple pathogens were more 

common in HM herds. Cryptosporidium spp. tended (p=0.06) to 

occur more often in HM herds. Another tendency was that the first 

pen in which calves were housed after the calving pen was cleaned 

more often in LM herds (p=0.09). HM herds treated diarrhoea with 

antibiotics significantly more often (p=0.002). Failure of passive 

transfer (total protein < 55 g/l) occurred significantly more often in 

the HM herds (p=0.02). LM herds started to feed calves with 

concentrates earlier (p=0.005) and tended to offer pelleted 

commercial calf starter more often (p=0.058). There was also a trend 

(p=0.07) that LM herds offered calves roughage earlier in life. The 

results indicate that gastrointestinal disorders may be an important 

cause of calf mortality in large Swedish dairy herds and that 

improved colostral management, hygiene, and feeding routines, and 

a controlled use of antibiotics may reduce the problems. 

Key words: calf mortality, diarrhoea, management 

223 Rapid Assay Evaluation to Detect Different Enteropathogens 

in Calf Faeces 

D. Klein, A. Kern, G. Lapan, V. Benetka, K. Möstl, A. Hassl, W. 

Baumgartner 

University of Veterianary Medicine, Clinic of Ruminants, Vienna, 

Austria 

Objective of study: The different aetiologies of calf diarrhoea 

without specific symptoms make the diagnosis and consequently an 

appropriate treatment and prevention difficult for veterinarians. 

Conventional diagnostic methods to detect the most important 

enteropathogens in diarrhoeic calves require time, experience and 

special laboratory equipment. Therefore rapid assays for detection of 

bovine coronavirus, rotavirus A, Cryptosporidium parvum and 

Escherichia coli F5 have been developed.These rapid 

immunomigration/immunochromatographic assays have been 

evaluated in the field. 

Material and Methods: Faecal samples of diarrhoeic and healthy 

neighbour calves from randomly chosen farms in Austria were 

included in this study. 

Results: Rapid tests for the detection of bovine coronavirus and 

rotavirus showed a high specificity but a relatively low sensitivity 

compared to RT-PCR. Sensitivity and specificity for detection of C. 

parvum were high. All results of the E. coli rapid assay were in 

accordance with bacteriological cultivation. 

Conclusion: The C. parvum and E. coli F5 test showed an excellent 

specificity and sensitivity and can be recommended to the practising 

veterinarian. Because of limited sensitivity the fast tests for the examined 

viruses are useful rather on herd level than in single individuals. 

224 Comparison of Different Strategies for the Therapy of 

Acidosis in Calves with Neonatal Diarrhea 

I. Lorenz 1 , G. Haase 2 , W. Klee 2 

2 University College Dublin, School of Agriculture, Food Science and 


2 University of Munich, Clinic for ruminants, Munich, Germany 

The aim of this prospective study was to determine, if and to which 

extent differences on a clinical and laboratory diagnostic level exist 

between rapid versus slow correction of metabolic acidosis in calves 

with neonatal diarrhea. Among the calves admitted to the Clinic for 

Ruminants between February, 2004, and March, 2005, 82 calves up 

to 3 weeks of age were included in this study. Further selection 

criteria were diarrhea, and metabolic acidosis with a base excess 

between -15 and -26 mmol/l. Exclusion criteria were hypoglycemia, 

hyperkalemia, and illnesses that had an influence on posture or 

behavior. According to the formula bodyweight x base deficit x 0.6 

the calves of both groups were given the respective amount of 

sodium bicarbonate, either within one hour as a 4.2 % solution, 

whereupon 0.9 % sodium chloride was given during the following 23 

hours in volumes corresponding to the estimated fluid loss, or as 


constant drip infusion over 24 hours which contained both the 

calculated amount of bicarbonate as a 1.4 % solution and 0.9 % 

sodium chloride which was estimated on the basis of clinical 

dehydration.At the initial examination and after the treatment by 

infusion on the second day careful attention was paid to posture, 

behavior, sucking reflex, palpebral reflex and tension of the tongue. 

Blood samples were taken upon admission of the animals and after 

completion of the fluid therapy on the second day. Calves in which 

acidosis was corrected rapidly showed statistically significantly 

lower pH, base excesses and HCO 3 - concentrations on the second 

day compared to the calves of the other group; but these variations 

were not clinically relevant. Thus no differences between the two 

groups could be diagnosed by taking into consideration all the 

clinical parameters recorded. Likewise, no differences between 

groups with regard to milk intake after the end of fluid therapy could 

be detected. Especially calves that were quickly compensated were 

often underdosed with HCO 3 - by using the dosage factor 0.6. 

Nevertheless, in practice, it can be recommended to treat calves with 

an initial rapid bicarbonate infusion to insure correction of acidosis, 

and thus amelioration of the general condition. In order to avoid t 

relapses, calves should be treated subsequently with additional 

amounts of bicarbonate or other buffer substances intravenously or 

orally according to dehydration status and sucking behavior. 

Key words: calf diarrhea, metabolic acidosis, therapy, sodium 

bicarbonate 

268 Efficacy of Metaphylactic Florfenicol Therapy during Natural 

Outbreaks of Bovine Respiratory Disease 

B. Catry 1 , L. Duchateau 2 , J. Van de Ven 4 , G. Opsomer 1 , 

F. Haesebrouck 3 , A. De Kruif 1 


Obstetrics, Reproduction and Herd Health, Merelbeke, Belgium 


Physiology and Biometrics, Merelbeke, Belgium 


Pathology, Baceriology and Poultry Diseases, Merelbeke, Belgium 

4 Schering-Plough Animal Health, Brussels, Belgium 

The efficacy of an injectable formulation of florfenicol (Nuflor ® , 

Schering-Plough) as metaphylactic control of naturally occurring 

BRD was evaluated in two double blind randomly controlled field 

studies on two Dutch veal calf herds (A and B). he number of 

animals enrolled in trial A and B were 351 and 344 respectively. 

Cattle in the direct environment of calves with clinical respiratory 

disease were randomly allocated to treatment with 40 mg/kg 

florfenicol, a positive control treatment (12.5 mg/kg tilmicosin p.o. 

BID for 5 consecutive days in herd A, and 12.5 mg/kg doxycycline 

p.o. BID for 5 consecutive days in herd B), or a negative control (1 

placebo saline administration on D0). n both trial A and B, a 

statistical difference (P

269 Evaluation of the Role of Clostridium perfringens Type A in 

Bovine Neonatal Diarrhoea 

C. Manteca 1 , G. Czaplicki 2 , A. Ginter 3 , J. Bughin 4 


2 ARSIA, Animal Health Department, Loncin, Belgium 

3 Bio X, R&D, Jemelle, Belgium 

4 ARSIA, Animal Health Department, Ciney, Belgium 

Objectives of the study: Cl. perfringens type A is today well known to 

induce enterotoxaemia in cattle. This study tries to highlight the 

putative role of alpha toxin produced by Cl. perfringens type A in acute 

enteritis of neonatal calves. To reach this goal, prevalence in diarrheic 

calves, macroscopic and histological lesions and results of an oral 

inoculation have to be described. 

Materials and Methods: Seventy one diarrheic and 249 normal faecal 

samples from BB and Holstein calves in Belgium and in France were 

examined by ELISA for the presence of alpha toxin, rotavirus, 

coronavirus, Cryptosporidium spp. Classic aerobic analysis and 

agglutination tests (F5, F17, CS31A) were performed to check the 

presence of potential pathogenic E. coli 

and BVD presence was checked by IFA. Necropsy and histology cases 

have been performed to precise enteric lesions. Diarrhoea apparition 

after oral inoculation of a calf with a toxigenic Cl. perfringens type A 

strain in liquid media have been compared with symptoms in a calf 

inoculated with broth only. 

Results: Alpha toxin was detected in 7 (10%) diarrheic calves and in 2 

(0,8%) normal calves. In diarrheic animals, pathogenic E. coli (51%), 

Cryptosporidium spp. (32%), Rotavirus (27%), Coronavirus (15%) and 

BVD (3%) were also detected. None Salmonella spp. have been 

isolated. Alpha toxin was detected without any other enteric pathogen 

in 2 cases. In these last cases, necropsy showed mainly an acute jejunoileitis 

with a yellow mucoid content, a hemorrhagic aspect of serosa 

and a local acute and congestive adenitis. Histological lesions seem 

mainly consist in a superficial desquamation of mucosa. Calf 

inoculated with toxigenic Cl. perfringens type A showed a catarrhal 

diarrhoea and uninoculated calf didn’t. 

Conclusions: Alpha toxin could be a frequent aetiology of neonatal 

diarrhea in calves. This pathogenic role of alpha toxin is today 

already well known in swine. In this species, we have showed that 

vaccination against alpha toxin and a specific antibiotherapy is able 

to improve considerably the clinical situation. More studies must be 

carried out to confirm this conclusion and to check efficacy of 

vaccination in cattle. 

Key words: Clostridium perfringens, alpha toxin, enteritis, calf 

270 Neonatal Calf Diarrhoea in Problem Farms Solved in 5 Easy, 

Simple and Cost Effective Steps 

J. Van De Ven 

Schering Plough Animal Health, Brussels, Belgium 

On 27 farms in the Netherlands with neonatal calf diarrhoea a simple 

diagnostic plan was implemented to help pinpoint the mayor infectious 

and management causes. Inclusion criteria for farms was a minimum 

prevalence of 50% diarrhoea among calves of 0-21 days of age. Step 1: 

Anamnesis Step 2: Faecal examination for rota/coronavirus, E.coli K99 

and C.parvum was done using on farm dipsticks (Biox Diagnostics, 

Belgium). Step 3: The transfer of passive immunity was evaluated by 

determining the IgG values in serum (GD, Netherlands) of calves 2-7 

days old (n=5/farm). Step 4: The colostrums quality of cows 

(n=20/farm) was measured cow-side using a colostrum density meter 

(CER Marloie, Belgium). 

Step 5: The effect of vaccination with an E.coli/rota/corona vaccine 

(Rotavec ® corona, Schering-Plough) was investigated by vaccinating 

all cows during a 2,5 month period. 

C. parvum was the most diagnosed pathogen on 51,85% of the 

farms. On 30 % of the farms rota/coronavirus or E. coli or a 

combination of these 3 pathogens was identified and on 50 % of the 

farms one or more of the 3 mentioned pathogens were found in 

combination with C. parvum. On 18,18% of the farms the average 

colostrum quality of all cows was less than 50 g/l. On only 9,09% of 

the farms the average colostrums quality was >100 g/l. The average 

IgG found in serum of calves was 9,62 g/l. On 18% and 61% of the 

farms the average IgG value was < 5 g/l and 40.0°C, and abnormal respiratory pattern, and 

depression), the animal was treated with a single sc injection of 2ml 

Resflor®/15kg bodyweight, providing 40mg/kg florfenicol and 2.2 

mg/kg flunixin. The rectal temperatures of the patients were 

measured again after 3, 6, 12, and 24 hours, and after 3 and 14 days. 

After 1, 3, and 14 days the clinical examination was repeated. The 

intake of milk replacer being offered twice daily was assessed for 4 

consecutive days and the daily weight gain was calculated using the 

individual body weights on day of arrival and 21 days later. 50 

animals fulfilled the inclusion criteria and were treated with 

Resflor ® . 30 animals stayed healthy and were used as negative 

control for the criteria milk replacer intake and weight gain. The 

reduced intake of the milk replacer offered of about 91.5% (SD 22.9) 

in sick animals returned to values as in healthy animals (98% to 

100%) until the subsequent feeding. There was no reduction in daily 

weight gain in treated animals (390g, SD 291) compared to the 

healthy animals (365g, SD 307). A rapid decrease of the initial rectal 

temperature of 40.48 °C (SD 0.35) to 39.39 °C (SD 0.40) after 3 

hours was observed. This temperature stayed between 38.62 °C and 

39.1 °C (SD 0.44 and 0.43) during the observation period. The 

depression observed in all animals at enrolment improved to 8%, 

0%, and 9% of the patients after 1, 3, and 14 days. Respiratory signs 

were observed in all animals at inclusion, and in 10%; 0%, and 9% 

of the animals after 1, 3, and 14 days. It could be demonstrated that 

a combined antibiotic and anti-inflammatory treatment leads to a 

rapid improvement of pyrexia, and a rapid return to normal feed 

intake resulting in non-inferior body weight gains compared to 

healthy animals. 

272 Intranasal Followed by Systemic Vaccination is an Optimal 

Vaccination Schedule for Young Calves against Bovine 

Respiratory Syncytial Virus and Parainfluenza Type 3 Virus 

I. Vangeel, R. Raue 

IgG low IgG High 

Rota Use Colostrum meter Housing Hygiene/ 

E. coli Implementation of Disinfection 

Corona Colostrum bank Temporary vaccination 

Vaccination (Vaccination in high Risk 

periods=Peak 

calving periods) 

Crypto Use Colostrum meter Halofuginone 

Implementation of Day 0–7 after birth 

Colostrum bank once daily 

Halofuginone Hygiene/Disinfection 

Day 0–7 after birth once daily Strict individual housing 

Hygiene/Disinfection 

Strict individual housing 

Both Combination of above Combination of above 

categories 

Calf Physiology, Management and Diseases 225

Pfizer Animal Health, Veterinary Medicine Research and 

Development, Sandwich, United Kingdom 

Outbreaks of bovine respiratory disease most frequently occur in 

young animals. Due to the high prevalence of maternal antibodies 

against Bovine Respiratory Syncytial Virus (BRSV) and 

Parainfluenza Type 3 Virus (PI3V), systemic two-dose vaccination 

of calves below three months of age often induces insufficient 

protection against these viruses. In this study, a vaccination 

schedule comprising intranasal vaccination with a modified live 

BRSV and PI3V vaccine at three weeks of age followed by 

systemic booster vaccinations with modified live vaccine at three 

and four months of age was evaluated in calves with and without 

maternal antibodies. Calves were either vaccinated intranasally 

(Rispoval ® RS+PI3 IntraNasal, Pfizer Ltd) at three weeks of age 

followed by booster vaccinations intramuscularly (Rispoval ® 3, 

Pfizer Ltd) at three and four months of age, vaccinated intranasally 

at three weeks of age only, vaccinated intramuscularly at three and 

four months of age only or not vaccinated at all. The same 

experiment was carried out in calves with and without maternal 

antibodies. Blood samples were collected weekly until four weeks 

after the second booster and BRSV and PI3V serum antibody levels 

measured by ELISA. In calves without maternal antibodies 

intranasal vaccination induced high serum antibody levels to 

BRSV and PI3V. The booster injections were necessary to prevent 

antibody levels for PI3V declining from nine weeks and BRSV 

from 14 weeks after intranasal vaccination as observed in calves 

that did not receive the boosters. Antibody responses to both BRSV 

and PI3V after the first intramuscular vaccination at three months 

of age were higher in calves vaccinated earlier with the intranasal 

vaccine compared with calves that only received their first 

vaccination at three months of age. In calves with maternal 

antibodies there was no increase in BRSV or PI3V serum antibody 

levels after intranasal vaccination. However, after the first booster 

again higher antibody levels were measured in calves earlier 

vaccinated intranasally compared with calves that had not been 

vaccinated before. BRSV and PI3V antibody levels of calves that 

were not given booster vaccinations continued to decline beyond 

three months of age. Based on the serological monitoring carried 

out in this study, it was concluded that intranasal followed by 

systemic vaccination is an optimal vaccination schedule for young 

calves against BRSV and PI3V. 

273 Case Control Study for the Evaluation of Risk Factors 

Associated with Infectious Respiratory Diseases in Dairy 

Calves in Québec 

D. Francoz 1 , G. Fecteau 1 , M. Fortin 2 , AM. Christen 3 , J. Arsenault 1 

1 

Université de Montréal, Clinical Sciences, St-Hyacinthe, Canada 

2 

Laboratoire d’Épimiosurveillance Animale du Québec, Clinical 

Sciences, Saint-Hyacinthe, Canada 

3 

Fédération des Producteurs de Bovins du Québec, Longueil, 

Canada 

A case control study was performed to evaluate the risk factors 

associated with infectious respiratory diseases in dairy calves in 

Québec.Twenty eight herds were selected as control based on 

absence of respiratory diseases in calves during the last 24 months. 

Fourteen herds were selected as case based on the history of 

endemic respiratory problems. All herds were visited twice at 1 

month interval. In all herds, 5 calves of less than 6 month of age 

were randomly selected. Blood samples were taken to evaluate IBR, 

BRSV, PI-3, BVD and Mycoplasma bovis for serology. Nasal swabs 

were preformed during the 1st visit for viral isolation, mycoplasma 

and routine bacterial culture. Milk bulk tank was sampled twice for 

mycoplasma culture. A questionnaire on herd practice was 

submitted to the herd manager and veterinarian. Statistical analysis 

was performed for the following risk factors: viral, bacterial and 

mycoplasma status, herd size, calf feeding practice, calves 

vaccination program and contact between animals. Exact 

conditional logistic regression stratified for the matched pair was 

used. All factors were tested in univariate analysis. Variables with 

P

Materials and Methods: 90 healthy naturally born calves, 45 Nelore and 

45 Limousin, were grouped according to their dam’s parity. Venous blood 

samples were collected from the 90 calves within the first 24 to 36 hours 

of life, and at 15, 30, 60, 90, and 120 days post-natal. 

Samples Results: The parity of the dam had no effect on mean 

values of serum gammaglobulins, serum and plasma proteins, and 

serum activity of gamma-glutamyl transferase (GGT) in calves 

within 24 to 36 hours of life. Although the serum gammaglobulins 

did not differ significantly between breeds, Nelore calves showed 

slightly higher serum and plasma proteins, IgG, significantly higher 

GGT (p

Key words: calf, active immunity, passive immunity, colostrum, 

vaccination 

348 Effects of Gradual Weaning and Transport on Leukocyte 

Antioxidant Enzymes and Plasma Malondialdehyde 

N. Burke 1 , G. Scaglia 2 , H. Boland 2 , W. Swecker, Jr 1 

1 

Virginia-Maryland Regional College of Veterinary Medicine, Large 

Animal Clinical Sciences, Blacksburg, United States 

2 

Virginia Tech, Animal and Poultry Sciences, Blacksburg, United 

States 

Weaning and transport of calves are stressful events associated with 

increased susceptibility to infectious disease and injury. Oxidative 

stress may exacerbate psychological and physiological demands 

brought about by these practices. Management strategies employing 

two-stage weaning can alleviate exhibition of stress-linked 

behaviors. Therefore, a trial was conducted to asses the effects of 

two-stage weaning and subsequent transport on oxidative stress 

markers in calves. Thirty-six crossbred Angus steers (243 ± 21 kg; 

221 ± 20 d) were blocked by weight and allotted to control (C), 

fenceline (FL) and nose clip (NC) groups. On day -7 of the trial FL 

calves were separated from dams by a fence, and NC calves were 

fitted with anti-suckle devices. Control and NC calves remained with 

dams until day 0, at which point all calves were permanently 

removed from dams and transported 172 km. Calves were weighed 

and bled by jugular venipuncture on days -7, 0 (pre-transit), 1 and 

+7. Leukocytes were analyzed for glutathione peroxidase and 

glutathione reductase activity, whereas the concentration of 

malondialdehyde (MDA) and Se was measured in the plasma. Data 

were analyzed using PROC MIXED with the Tukey-Kramer 

adjustment applied post-hoc. No treatment effects were detected for 

blood variables or calf weights. Glutathione reductase activity 

declined (P

Table 2. Ratio of different stomachs in comparison with the total 

stomach volume of dairy calves (%) 

calf age (weeks) ruminoreticulum abomasum omasum 

4 86 13 1 

8 80 17 3 

13 87 8 5 

development considering size, localisation or pathological alterations. 

Measurements are possible without influencing viability of animals. 

352 Medical and Organizational Challenges Associated with the 

Birth of Cloned Calves 

G. Fecteau 1 , AC. Brisville 1 , S. Boysen 1 , S. Buczinski 1 , P. Dorval 1 , 

R. Lefvebre 1 , P. Hélie 2 , A. Desrochers 1 , L. Smith 3 

1 

Université de Montréal, Sciences Cliniques, Saint-Hyacinthe, 

Canada 

2 

Université de Montréal, Pathologie et Microbiologie, Saint- 

Hyacinthe, Canada 

3 

Université de Montréal, Sciences Biomédicales, Saint-Hyacinthe, 

Canada 

Somatic cell nuclear transfer (SCNT) is a reproductive biotechnology 

with a broad spectrum of applications, however the cloning process 

remains relatively inefficient. In cattle, the success rate is about 5 % 

and depends mainly on IVF culture system, cell-line and genotype. 

Due to the high risks of cloned pregnancies, the teaching hospital at 

the Université de Montréal developed a multidisciplinary critical 

care unit to monitor newborn calves. Clinicians and residents, in 

conjunction with specially trained animal health technicians and 

veterinary students provided 24 hour care. A data recording system 

was developed to facilitate monitoring. Between 2004 and 2007, 

twenty-six Holstein cloned calves were delivered by caesarean after 

induction between 270 and 280 days of gestation. Birth weights 

varied from 35 to 82 kg (median 55 kg, mean 56 kg ± 8.9). Clinical 

evaluation included intrauterine foetal assessment, monitoring of the 

cows’ condition and a complete assessment of the newborn calves 

(physical examination, blood count (CBC), serum biochemistry 

profile, thoracic radiograph and arterial and/or venous blood gas 

analysis). Hypoxemia (PaO 2 = 30.4-79.6 mmHg, median = 4 

8mmHg), associated with hypercapnia (PaCO 2 = 55.7-89.3 mmHg, 

median = 82.9mmHg), was commonly observed in the cloned calves. 

Other abnormalities included anaemia, azotemia and elevated body 

temperature. The precise aetiology of the respiratory dysfunction 

remains under investigation. Although many calves suffered from 

respiratory difficulties (22/26, 84.6%), only three died from 

respiratory failure (11.5%). Intranasal oxygen supplementation 

seemed to be an effective means of supporting calves suffering from 

hypoxemia without hypercapnia (success rate of 100%, 13/13). 

When hypoventilation was associated with hypoxia, mechanical 

ventilation appeared to be an interesting option (success rate of 

62,5%, 5/8). Finally, delivered calves had a survival rate of 69,2% 

(18/26) at 3 months of age. With a multidisciplinary approach, the 

teaching hospital successfully monitored and treated several cloned 

calves and developed a standardized protocol to manage high risk 

bovine pregnancies. 


992 Relationship between the Debfee of Dehydration and the 

Balance of Acid-Base Changes in Dehydrated Calves with 

Diarrhoea 

H. Guzelbektes, A. Coskun, I. Sen 

University of Selcuk, Faculty of Medicine, Department of Clinical 

Sciences, Faculty of Veterinary Medicine, Konya, Turkey 

Neonatal calf diarrhoea is a multifactorial disease which despite decades 

of research on the topic remains the most common cause of death in young 

calves. Diarrhea in calves can lead to dehydration, acidemia due to 

primarily to strong ion (metabolic) acidosis, hyperkalemia, and impaired 

cardiovascular and renal function. The objectives of study: The aim of the 

study was to investigate acid-base changes and to determine some serum 

biochemical parameters in dehydrated calves with diarrhoea according to 

the degree of dehydratation. 

Materials and methods: Thirty diarrhoeic dehydrated calves 

submitted to treatment in the university clinic, were used in the study. 

The calves were accessed as suitable for this study, if they were 

moderately or severely dehydrated according to clinical symptoms. All 

sick calves had the usual yellow and watery diarrhoea. Calves with 5% 

to 8% dehydration (moderate) had a weak suckle reflex, dry mucous 

membranes, warm mouth and partly good muscular tone. Calves with 

10% and above dehydration (severe) were unable to stand, and had no 

suckling reflex and cold mouth. 

Results: The mean pH, base excess, HCO 3 - , chloride (only severely 

dehydrated group) and sodium concentration were significantly 

decreased in both moderately and severely dehydrated group. On the 

other hand, potassium, phosphor and anion gap levels were increased 

compared to control group. 

Conclusion: There was relationship between base excess and anion 

gap with degree of clinical dehydration. However, there may not be 

an exact correlation between the degree of dehydration and the 

severity of acidosis. Based on the clinical symptoms, the results of 

this study could be a useful tool under field conditions in estimating 

the base excess in diarrhoeic dehydrated calves, when acidosis 

therapy is needed. 

Key words: calves, diarrhoea, dehydration, acid-base balance 

993 Epidemiology and Identification of Bovine Infectious and 

Tick-Borne Diseases in White Tailed Deer in Northern 

Mexico 

A. Cantu 1 , J. Ortega 2 , Z. Garcia 3 , H. Scott 2 , J. George 5 , J. Mosqueda 3 

1 

INIFAP- CIRNE, Epidemiology, Tampico, Mexico, Mexico 

2 

Texas A&M university kingsville, Epidemiology, kingsville, Texas, 

United States 

3 

INIFAP- CIRNE, parasitology, Mexico State, Mexico 

4 

Texas A&M university kingsville, Wildlife & animal science, 

kingsville, Texas, United States 

5 

USDA-ARS, parasitology, Kerrville, Tx., United States 

6 

INIFAP- CIRNE, Bioscience and Biotechnology, Mexico, Mexico 

7 

INIFAP- CIRNE, Wildlife & animal science, Mexico, Mexico 

The emergence of new and re-emergence of past pathogenic 

infectious diseases such as West Nile virus, brucellosis and tickborne 

diseases, may be linked to wildlife serving as reservoirs or 

vectors for spreading diseases. White-tailed deer (Odocoileus 

virginianus) are an important secondary host for the cattle fever tick 

species Rhipicephalus (Boophilus) annulatus and R. (B.) 

microplus. Due to habitat sympatry, deer may play a significant role 

in the transmission of bovine diseases. This project was designed to 

evaluate the prevalence of diseases and estimate risk factors 

associated with the transmission of infectious agents and tick borne 

diseases. The objectives were to determine the presence of bovine 

Babesia, brucellosis, leptospirosis, bovine viral diarrhea virus 

(BVDV), and infectious bovine rhinotracheitis (IBR) in white-tailed 

deer. A total of 457 whole blood samples were collected from whitetailed 

deer during the spring of 2004. Samples were tested for 

Babesia bovis and Babesia bigemina by nested PCR and indirect 

immunofluorescence antibody test (IFAT). Nineteen (4.2%) and 6 

(1.7%) samples were positive to B. bigemina and B. bovis, 

respectively. The positive blood samples to n-PCR were sequenced 

to demonstrate the specificity of the sequence of B. bovis (accession 

number M38218) and B. bigemina (accession number S45366). The 

results of the serological testing showed that 59.9 % (274) of deer 

sampled were positive to B. bovis and 5.5% (25) to B. bigemina 

antibodies. The prevalence for leptospirosis, IBR, and BVDV was of 

5.6%, 41.1%, and 63.5%, respectively. No positive samples were 

found to brucellosis. This is the first report of the presence of B. 

bovis and B. bigemina in white-tailed deer. This finding may suggest 

that white-tailed deer may act as a reservoir for two bovine Babesia 

species, and may be a source of transmission for new or reemergence 

of bovine diseases to cattle or another wildlife species. 

994 Evaluation of Diarrhoea Treatment with Injectable 

Enrofloxacin (Baytril® 5% Bayer) and Oral Rehydration 

Solution (Glutellac® Bayer) in Newborn Lactating Calves 

V. Cuteri 1 , L. Nisoli 2 , A. Fruganti 1 , A. Romero Tejeda 1 , A. Attili 1 


1 University of Camerino, Veterinary Science, Matelica (MC), Italy 

2 Bayer Health Care, Animal Health Division, Milan, Italy 

Diarrhoea is a common clinical disease in newborn lactating calves, 

causing a high mortality. As fluid and electrolyte loss induced by 

diarrhoea results in dehydration and metabolic acidosis, a major goal 

of treatment is also administration of electrolyte solutions in order to 

restore the intestinal equilibrium. To evaluate therapeutic effects of 

an oral dietary alkaline supplement for rehydration (Glutellac ® - 

Bayer), a multi-centre and randomised clinical study was carried out 

on 79 dehydrated diarrheic calves aged 3-70 days, resulted 

bacteriologically positive to Escherichia coli sensitive to 

Enrofloxacin. All animals were treated with Baytril ® 5% - Bayer 

(enrofloxacin: 0,5 ml/10 kg of b.w./s.c.). 40 calves (group A) 

furthermore received Glutellac ® (50 ml in two litre of water twice 

daily), administered orally for 3 consecutives days, while thirty 9 

calves (group B) didn’t receive oral electrolyte solution. The clinical 

examinations of all calves were performed at the 1 (first day of 

treatment), 2, 3, 4, 5 and 10 days post treatment (PT). The following 

parameters: rectal temperature, general health, faecal consistency, 

dehydration, general conditions at day 5 and other episodes of 

diarrhoea appears into 30 days from the inclusion, were recorded. In 

comparison with group B, a significant rate (95.0%) of calves treated 

with the association of Baytril ® 5% and Glutellac ® showed a shorter 

rehydration time already at 4 th day PT (P=0.0178). At the same time 

(4 th day), the Glutellac ® treatment increased the number of animals 

whit normal general health (87.5% vs 52.6% of group B: P= 0.0007), 

and improved the faecal consistency in the 82.5% of calves in 

comparison to the 50.0% of calves belonging to group B (P=0.0023). 

Besides, it was observed a significant difference about the total 

clinical conditions recorded at 5 days PT between groups; the 

clinical recovery concerned the 95.0% of animals of group A and 

only the 78.9% of calves of group B (P=0.034). Glutellac ® replaced 

fluid, replenishes lost electrolytes, rapidly compensates metabolic 

acidosis and supplies vital energy. Unlike other buffers, the buffer 

substance acetate does not interfere with the casein precipitation in 

calf’s abomasum and with the milk digestion, promoting 

regeneration of the gut epithelium. This study indicated that 

administration of Baytril ® 5% associated with Glutellac ® resulted as 

quicker, practical, economical and effective way for the treatment of 

dehydrated diarrhoeic calves. 

995 Effectiveness of Sodium Acetate in Neonatal Calf Diarrhoea 

I. Zhirkov 

Volgograd State Agricultural Academy, Veterinary Medicine, 

Volgograd, Russian Federation 

Newborn calf may deal with different situations requiring digestive 

adaptations. Usually digestive disorders in neonatal animals result in 

diarrhoea. In Russia often technological stresses (infringements of 

feeding and rearing regime, hygiene, etc.) of dairy breeds cause 100 

% morbidity of neonatal calves. In the alarm phase the sympathoadrenal 

system of organism is excited resulting in blocking of 

parietal cells secretory activity in the abomasum. Colostrum poorly 

processed and inseminated with environmental microflora is 

emptied into duodenum. Specifically, E. coli bacterial numbers are 

increased 5- to 10,000- fold in the small bowel of calves with 

naturally acquired diarrhoea. Our physiological experiments on 

operated calves with 2-shaped duodenal cannulae showed diarrhoeic 

calves to have achlorhydria, abomasal chyme pH 5.0 - 6.0 

(measurements were made every 2 h. during the 24 h.). We also 

reported earlier acetate-ion to be a potent stimulant of abomasal HCl 

secretion. Field trials were performed in the dairy farm “Luch” Ltd. 

286 healthy-born Holstein calves were observed since the birth (BW 

34.1±4.7 kg) up to 30 days. Just after calving neonates were kept 

separately. All calves were randomly divided to three groups: two 

experimental and a control. Diarrhoeic calves of the control group 

(CG) were treated symptomatically using the antibiotics (neomycin 

IM, chloramphenicol and chlortetraciclin PO). Instead of colostrum 

(milk) they received isotonic saline. In case of diarrhoea animals of 

the experimental group I (EG1) were treated according to the 

following scheme: – one meal (just after appearance of diarrhoeic 

symptoms) was missed, instead of colostrum sick calf was fed the 

same volume of saline; – next meal sick calf was given 1L of 3 % 

sodium acetate aqueous solution (SAAS); – sodium acetate was 

added to colostrum (milk) in the dose of 20 g/L for two days. 


Calves of the experimental group II (EG2) were prophylacted the 

incidence of diarrhoea. During 2 - 10 days of life each animal was 

given 1L of 1 % SAAS in an hour after prandial feeding. In the case of 

diarrhoea, calves of the EG2 were treated with 3 % SAAS according to 

the scheme designed for EG1. Results are presented below. 

Group n Morbi- Days Relapses BW in Morbi- Died, 

dity, of of 10 days dity % 

% illness/ diarrhoea old of 

100 / 100 respiratory 

calves calves diseases % 

Control 88 100 622.8 86.4 35.1±6.8 20.5 9.1 

Exper I 88 100 259.1 88.6 36.9±4.2 18.2 5.7 

Exper II 110 74.5 111.8 7.3 37.1±5.8 0 0 

996 Effect of Selenium and Vitamin E Supplementation during 

Late Pregnancy on Serum IgG Concentration in Heifers and 

Serum IgG Concentration and Passive Immunity in their 

Calves 

M. Moeini 1 , E. Mikaeili 1 , H. Karami 1 , A. Mostafaie 2 

1 Razi University, Animal Science, Kermanshah, Iran 

2 Razi University, Microbiology, Kermanshah, Iran 

Fifty Holstein heifers were randomly assigned to one of five 

treatments. Four and two weeks before expected calving, heifers 

were injected intramuscularly 10 ml (T1), 20 ml (T2), 30 ml (T3), 40 

ml (T4) selenium and vitamin E respectively. The control (C) group 

received no supplement.Blood samples were collected from heifers 

four weeks before expected calving and at calving day. Blood 

samples of calves were drowning from the jugular vein at birth and 7 

days of age. White blood cell and differential leukocyte counts were 

measured. Serum concentrations of Se were determined using 

hydride generation Atomic Absorption Spectrophotometery. Serum 

IgG concentrations were measured by Sandwich ELISA method. 

The result indicated that Serum Se concentrations of heifers before 

injection of Se and VE supplement was not differ between 5 

groups(p>0.05), but after calving in T3 and T4 being higher 

(p0.05). The 

changes of mean values of serum IgG did not differ among calves. 

The white blood cell counts were higher in calves from T4 group 

compare to control group (p0.05). 

Key word: selenium, vitamin E, IgG, immunity, heifers, calves 

997 Clinical Case Report of Congenital Abnormalities in Tow 

Holstein Calves with Spina Bifida Occulta, Tail Absence, 

Achondroplasia and Brachygnatia 

F. Zavoshti 1 , M. Dezfouli 2 , M. Tooloei 1 


Clinical Sciences, Tabriz, Iran 

2 University of Tehran, Department of Clinical Sciences, Tehran, Iran 

Objectives of the study: Clinical case report of congenital 

abnormalities in tow Holstein calves in suburbs of Tehran, Iran. 

Materials and methods: Two Holstein calves were examined at 

2005 in a large dairy farm in suburbs of Tehran province, Iran. One 

of calves was a 47 days old and weighting 35.5 Kg at the time of 

examination. Height of calf was 72 cm from whiter to hoof. Signs 

such as tail absence (Hypoplastic coccyx) and spina bifida occulta 

were prominent on lumbosacral area. Presence of skin and hair coat 

at the area didn’t permit neural structure of lumbar vertebrate to be 

exposed. Apart from the clinical signs of the calf the calving was 

normal. Other calf was an 11 month old female calf that was 

examined the same time because of respiratory disorders. In clinical 

evaluation brachygnatia inferior (mandibular) was observed. Further

clinical examination did not show other congenital abnormalities 

such as hair lip and Clift palate. 

Conclusions: In a case we can find more than one congenital 

abnormalities such as case former, also although some congenital 

abnormalities not-life threatening, is usually considered an important 

defect if detected during soundness or respiratory examination such as 

second case. 

Key words: congenital abnormalities, tail absence, spina bifida 

occulta, achondroplasia, brachygnathia, calves 

998 Diarrhea of Neonatal Calves due to Klebsiella (First Report 

from Iran) 

F. Zavoshti 1 , M. Dezfouli 2 , A. Bahonar 3 , M. Rabbani 4 , T. Salehi 4 






of Food Hygiene, Tehran, Iran 


of Microbiology, Tehran, Iran 

Objectives of the study: Clinical evaluation of Klebsiella induced 

diarrhea and frequency of this agent in the diarrhea of calves in Iran. 

Materials and Methods: Procedure was collecting stool sample 

from rectum of the all affected and apparently healthy calves in a 

cross sectional study and using bacteriological standard methods for 

detection of this bacterium. Statistical analysis was SPSS package. 

Results: 31 samples out of 209 diarrheic calves (lees than one month 

age) was examined stool examination that was infected with 

Klebsiella. In this regard, 18 samples (8.6%) were from diarrheic 

calves and 13 samples (15.3%) from control group. Mean of age in 

diarrheic calves was 12.00±3.08 days. In diarrheic group 6 calves 

(33.3%) showed severe diarrhea. All diarrheic calves (100%) 

showed thin and vivacity in general condition. Nine diarrheic calves 

(50%) had yellow feces. 17 of diarrheic calves (94.4%) had poor 

feeding behavior and all diarrheic calves (100%) showed second 

dehydration degree. 

Conclusions: The clinical implication is Klebsiella can be one of 

causative agents in the calves’ diarrhea syndrome in the less than one 

month age. 

Key words: Klebsiella, diarrhea, calf, Iran 

999 Clinical Trial of Different Type of Dried Colostrum Powders 

in Neonatal Holstein Calves to Prevent Diarrhea in Iran 

F. Zavoshti 1 , M. Dezfouli 2 , M. Rabbani 3 , T. Salehi 3 , H. Seifi 4 

1 

Faculty of Veterinary Medicine, University of Tabriz, Department of 


2 

Faculty of Veterinary Medicine, University of Tehran, Department 


3 


of Microbiology, Tehran, Iran 

4 

Ferdowsi University of Mashhad, Department of Clinical Sciences, 

Mashhad, Iran 

Objectives of the study: Transfer of passive immunity in newborn 

calves occurs through the oral consumption and subsequent 

absorption of immunoglobulin (Ig) soon after birth. Prevalence of 

failure of passive transfer (FPT) in calves has been reported to range 

from 11 to 31% in North America. Many attempts have been made to 

artificially augment the ability of calves to attain passive immune 

support, such as stored surplus colostrum, injectable Ig solutions, 

dried colostrum and concentrated milk whey. However, none of 

these methods have proven to provide adequate passive immune 

support. The aim of this study was to test the value of lyophilized 

and spray-dried colostrum as a colostrum supplement on the 

incidence of neonatal calf diarrhea. 

Materials and Methods: One hundred and twenty healthy Holstein 

calves were assigned randomly at birth to one of four groups; each 

group contained 30 calves and the calves were received as follows: 

group A, 85± 5 g of lyophilized colostrum powder dissolved in 3 kg 

of whole milk; group B, 85± 5 g of spray- dried colostrum powder 

dissolved in 3 kg of whole milk; group C, 750 ml frozen and thawed 

colostrum in 3 kg of whole milk, and group D, only 3 kg of whole 

milk without colostrum supplement. Each group was subdivided into 

3 subgroups of 10 calves as follows: calves fed colostrum 

supplement at 24 or 48 or 72 h after birth. All calves fed fresh 

colostum within 6 h after birth at the amount of 5% BW by bottle. 

Results: Calves which were fed lyophilized colostrum supplement had 

a significant lower incidence of diarrhea in comparison to control 

calves. In addition, calves were received lyophilized colostrum 

supplement at 24 h after birth showed less incidence of diarrhea than 

calves fed supplement at 48 and 72. 

Conclusions: According to the results of present study providing 

lyophilized colostrum supplement at 24 h after birth might have been 

preventive effect on calf diarrhea. The spray- dried colostrum 

supplement had not the same effect. 

Key words: calf, diarrhea, colostrum, lyophilized, spray-dried 

1000 The Importance of Acute Phase Protein Measurements in 

the Clinical Diagnosis of Calves Experimentally Induced 

with Lipopolisaccharide (E. coli) Endotoxemia 

A. Coskun, I. Sen 

University of Selcuk, Faculty of Vet. Med., Department of Internal 

Medicine, Konya, Turkey 

The objectives of this study was to determine the clinical changes, 

the importance of disseminant intravasculer coagulopathy and 

concentrations of acute phase proteins (Haptoglobin, fibrinogen and 

Serum Amilod A) in the diagnosis and prognosis of calves that were 

experimentally induced with Lipopolisaccharide (LPS) E. coli 

endotoxemia. 

Materials and methods: a total of eight healthy Holstein calves, 

aging 30 to 45 days old were used in this study. In all calves, the 

endotoxemia were induced by giving 0,1Ïg/kg doses of LPS in 50 ml 

phisyologic saline via intra-venous route in constant infusion during 

30 minutes. Blood samples were collected before (24 hour) and after 

the endotoxemia induction at 0, 0.5 th, 1, 2, 4, 6, 8, 12, 18, 24, 48, 72, 

96, 120, 144, 168, 192, 216 and 240th hours of study. In addition, 

clinical symptoms of endotoxemia were also observed following 

LPS infusion. 

Results: in general, abnormal clinical changes such as anorexa, 

depression, increased respiratory and heart rate were observed 30 

minutes later following LPS infusion. Haemotological examination 

showed that a significant leucopenia, folowed by a leucousytosis and 

decresead T and B-Lyphocytes levels were detected at the first hours of 

study. Abnormal chances were also observed in cougulation 

parameters such as APTT, PT, trombosit and fibrinogen during 

study.The plasma Haptoglobin (Hp) concentrations were increased 

from basal levels (8,13 µg/ml) to 20,84µg/ml levels at 6 th hours 

following LPS infusion. The highest Hp concentration (500,08 µg/ml) 

was found at 36 th hours after LPS infusion. And than, the Hp levels 

were declined to basal levels at 144 th hour of study. First 3 hours 

following LPS infusion, the Serum Amilod A (SAA) concentrations 

have increased from basal levels (12,04 µg/ml) to 30,22 µg/ml levels, 

and the highest concentration (185,27 µg/ml) of SAA was measured at 

24. at hours of study. And than SAA level has declined to basal levels 

at 120 th hours of study. 

Conclusion: based on clinical and laboratory findings, the 

endotoxemia was successfully induced by giving 0,1µg/kg doses LPS 

via intra-venous route in constant infusion. Results suggest that both 

Hp and SAA could be used as a indicator in the diagnosis of calves with 

endotoxemia. But, we could be say that haptoglobin might be a better 

prognostic indicator for calves with endotoxemia compare to SAA. 

This study was supported by both Tubitak and S.U. BAP 

Key words: calf, LPS, haptoglobin, SAA, endotoxemia 

1001 Protective Effect of Pastobov and Alterations of Peripheral 

Blood Leukocyte Subpopulations in Calves Experimentally 

Challenged with Mannheimia haemolytica A1 Leukotoxin 

D. Bednarek 1 , R. Urban-Chmiel 2 , K. Dudek 1 

1 National Veterinary Research Institute, Department of Cattle and 

Sheep Diseases, Pulawy, Poland 

2 Faculty of Veterinary Medicine University of Agriculture, 

Department of Veterinary Prevention, Institute of Infectious and 

Invasive Diseases, Lublin, Poland 

The aim of the study was to evaluate the effect of Mannheimia 

haemolytica A1 leukotoxin (LKT) on clinical status and alterations 

of cellular immune response in calves vaccinated (Group I) and non- 


vaccinated (Group II) with Pastobov (the vaccine manufactured by 

Merial consist of M. haemolytica A1 leukotoxoid). The alternations 

were investigated in peripheral blood leukocyte subpopulations 

using flow cytometry (FCM) in the calves experimentally challenged 

by intravenous administration of 25 µg of M. haemolytica A1 

leukotoxin per animal. The experimental calves (Group I) 21 days 

before the challenge were vaccinated with intramuscular injection of 

Pastobov (2 mls i.m.). The calves were next compared with the 

control animals (Group II - non-vaccinated). The comparing studies 

were performed just before (0) and 1, 2, 3, 4, 5, 6 and 24 hrs after the 

challenge. The following parameters were assayed: white blood cell 

count (WBC), percentage of polymorphonuclear leukocytes 

(PMNL), mid-size leukocytes (MID), i.e. total value of monocytes, 

eosinophils and basophils, total percentage of lymphocytes (LYM) 

and their subsets: CD2 + (T lymphocytes), CD4 + (T helper 

lymphocytes), CD8 + (T suppressor/cytotoxic lymphocytes) and 

WC4 + (B lymphocytes). A part from that the clinical signs were 

recorded during the study such as rectal temperature, heart and 

respiratory rate and general behaviour of the affected animals. The 

obtained results of the study showed, that in peripheral blood WBC 

and the percentages of MID, PMNL, LYM and their some 

subpopulations (CD2 + , CD4 + , CD8 + cells) markedly decreased 

(leukopenia) in both groups of the affected animals especially during 

the first of third hours of the experiment. However, these changes 

were more significant (P

faeces of healthy calves. Of the 37 E. coli isolated from diarrheic 

calves, K99 (18.9%), F41 (18.9%), heat-stable enterotoxin a (STa) 

(18.9%), Shiga toxin1 (Stx1; 13.5% ) and Shiga toxin2 (Stx2; 5.4%) 

and intimin (8.1%) genes were identified by multiplex PCR. Of the 

18 E. coli isolated from healthy calves, K99 (16.6 %) and intimin 

(55.5%) genes were identified by PCR. A total of 15 rotavirus, 11 

coronavirus and 11 E. coli K99 were detected in diarrheic calves by 

the antigen ELISA. 

Conclusion: This study shows that rotavirus, coronavirus, E. coli and 

Enterococcus ssp. were determined to play a role in the etiology of 

diarrhea in the neonatal calves. K99, F41, STa, Stx1 and Stx2 were 

found as the most common virulence gene markers of E. coli strains 

isolated from calves with diarrhea. The multiplex PCR may be useful 

for characterization of E. coli isolated from calves. 

This was supported by University of Selcuk, BAP. 

1005 Efficacy of Oral Rehydration Therapy Solutions Containing 

Sodium Bicarbonate or Sodium Acetate in Treating Calves 

with Naturally Acquired Diarrhoea and Moderate 

Dehydration 

I. Sen 1 , V. Altunok 2 , M. Ok 2 , P. Constable 3 , A. Coskun 1 

1 University of Selcuk, Faculty of Medicine, Clinical Sciences, 

Internal Medicine, Konya, Turkey 

2 University of Selcuk, Faculty of Vet. Med., Clinical Sciences, 

Internal Medicine, Konya, Turkey 

3 Pudue University, School of Veterinary Medicine, Veterinary 

Clinical Sciences, Indiana, United States 

The optimal formulation for an oral rehydration therapy (ORT) to treat 

calves with diarrhoea remains controversial. The objective of this study 

was to determine and compare the effect of four ORT solutions on 

indices of acid-base balance, abomasal emptying rate, and plasma 

volume expansion in cross bred calves with naturally acquired 

diarrhoea and moderate dehydration. 

Material and methods: Twenty calves up to 45 days of age with 5 

to 8% dehydration and mild acidaemia were enrolled in the study. 

Calves were randomly allocated to receive 2 L of one of four 

treatments (n = 5 each) administered at room temperature (20-22 °C) 

by oroesophageal intubation. The 4 treatments were 150 mmol/L 

sodium bicarbonate, 150 mmol/L sodium acetate, 300 mmol/L 

sodium bicarbonate, and 300 mmol/L sodium acetate. All 4 test 

solutions contained acetaminophen and 50 g of glucose 

monohydrate, equivalent to 1.3 g of glucose per kg of body weight. 

Jugular venous blood samples for determination of blood gas and 

serum biochemical parameters and acetaminophen concentration 

were obtained periodically before and after administration of the 

ORT solution. 

Results: The rate of systemic alkalinization, as assessed by change 

in venous measured strong ion difference and plasma bicarbonate 

concentration, was similar for all 4 treatments. Interestingly, jugular 

venous Pco 2 increased over the first 30 to 60 minutes in calves 

administered 300 mmol/L or 150 mmol/L sodium bicarbonate 

solution. For comparison, jugular venous Pco 2 decreased or stayed 

constant over the first 30 to 60 minutes in calves administered 300 

mmol/L or 150 mmol/L sodium acetate solution.150 mmol/L 

solutions were emptied faster from the abomasum than 300 mmol/L 

solutions, with no difference in emptying rate between acetate or 

bicarbonate containing solutions. 

Conclusion: Sodium acetate ORT solutions increase jugular venous 

pH and plasma bicarbonate concentrations at least as fast, and possibly 

faster, than sodium bicarbonate ORT solutions of similar osmolality. 

Although the rate of abomasal emptying was slowed when the ORT 

solution osmolality was increased, the slowing did not appear to have a 

clinically important effect of the rate of systemic alkalinization. Our 

results do not support widely held beliefs that bicarbonate containing 

ORT solutions provide the fastest correction of acid-base abnormalities 

in calves with diarrhoea and moderate dehydration. 

This study was supported by a grant from Tubitak and partly University 

of Selcuk, BAP 

Key words: calves, OES, acetaminophen 

1006 Acid-base Analyses of Venous Blood in Calves with Diarrhoea 

S. Canikli, A. Coskun, H. Guzelbektas, I. Sen 

University of Selcuk, Faculty of Medicine, Clinical sciences, internal 

medicine, Konya, Turkey 

Diarrhea is an important cause of morbidity and mortality in 

unweaned dairy heifer calves in most countries. Diarrhea in calves 

can lead to dehydration, acidemia due to primarily to strong ion 

(metabolic) acidosis, hyperkalemia, and impaired cardiovascular and 

renal function. Metabolic acidosis is well recognised potentially lifethreatening 

consequence of diarrhoea. It is due to the loss of 

bicarbonate into the small intestine and the production of acid in the 

colon. The objective of this study was to determine strong ion 

difference (SID), leukocyte, heamatocrit (Hct) and acid-base 

analysis of venous blood in calves with diarrhea. 

Material and methods: The pH, partial pressure of oxygen (pO 2), 

partial pressure of carbon dioxide (pCO 2), concentration of 

bicarbonate (HCO 3-), base excess (BE) and oxygen saturation (sat- 

O 2), anionic gap (AG), sodium (Na + ), potassium (K + ), cloride (CI – ), 

Strong Ion Differances, Hct, leukocyte and heamoglobin (Hgb) were 

measured in venous blood of 84 calves with diarrhea. After clinical 

examination, jugular venous blood samples were anaerobically 

obtained in a 3-mL plastic syringe that has been previously coated 

internally with sodium heparin. All sick calves had the usually 

yellow and watery diarrhoea. Sick calves had also similar clinical 

symptoms such as a weak suckle reflex, dry mucous membranes, 

unable to stand, and cold mouth. 

Results: The mean leukocyte count of calves with diarrhoea were 

significantly increased. The mean pH (7.16), BE (-12,5 mmol/L) , 

HCO 3- (13.33 mmol/L), Tco 2 (16.5 mmol/L), and oxygen saturation 

(36,28%) were significantly decreased compared to normal 

reference values. The mean levels of potassium (4.7 mmol/L) and 

anion gap (27.07) were also increased in calves with diarrhea. 

Conclusion, we could be said that the loss of bicarbonate in the 

feaces is regarded as cause of acidosis. Therefore, administration of 

sodium bicarbonate via intra venous due to significantly lowered pH 

and bicarbonate would be useful in treatmet of metabolic acidosis of 

calves with diarrhea. 

This study was supported by University of Selcuk, Scientific Research 

Project Coordinating 

Key words: calf, strong ion acidosis, pH, HCO 3 

1007 Detection of Five Agents (Coronavirus, Rotavirus, E. coli, 

Salmonella spp and Cryptosporidium sp) from Calves 

Diarrheic Samples in La Laguna Mexico Region 

J. Contreras, J. Palacios 

Schering Plough Animal Health, Animal Health Division, Mexico 

City, Mexico 

Introduction: Diarrhea is an important cause of mortality in calves and 

results in economic losses to the dairy industry 1, 2. Infected cows 

seldomly show clinical diarrhea, making it difficult to establish the 

carrier-state in the herd. Frequently identified causative pathogens are; 

E. coli, rotavirus most common during 4 to 14 days age, coronavirus 

more prevalent during 4-30 days age Cryptosporidium parvum from 1 

to 4 weeks age and Salmonella typhimurium and Dublin responsible for 

various clinical syndromes. Causative diagnosis is a critical tool in 

treatment and prevention. 

Study Objectives: To detect the presence of enteric pathogens (E. coli, 

Rotavirus, Coronavirus, Salmonella spp and Cryptosporidium sp) in 

calves with clinical diarrhea on 25 dairy operations from La Laguna 

region in Mexico. 

Material and Method: 170 feces samples from calves with diarrhea 

were aseptically collected during November and December 2006. 

Samples were analyzed by ELISA for Rotavirus, Coronavirus, and 

E. coli F5 using micro plates coated with monoclonal antibodies, 

conjugates with a plate reader with a 450 nm (Biovet Diagnostics) 

Samples were incubated in Salmonella enrichment media (Selenite 

broth) and plated on Green-brilliant media. Suspect colonies were 

further identified by biochemical methods. Cryptosporidium sp were 

detected by microscopic direct observation of fecal smears using 

Ziehl-Nielsen modified stain. 

Results: Samples were positive for Cryptosporidium sp, 99 cases 

(58, 2 %), Rotavirus, 54 (31.7 %), E. coli-F5, 33 (19.4 %), 

Coronavirus, 25 (14.7%) and Salmonella spp, 13 (7.65%). Mixed 

infections were detected in most samples with only 4.9% of the 

positive samples having only one antigen. There were no negative 

samples to any antigens most frequent antigen combinations were 

(Table 1). 

Discussion: The study confirms the presence of pathogens described in 

other world areas, from calves with clinical diarrhea, often as mixed 


# Antigens Association in diarrhea samples % 

4 Rotavirus-Coronavirus-E.coli-Cryptosporidium 39.7 

3 Rotavirus-E.coli-Cryptosporidium 14.3 

2 Rotavirus-Cryptosporidium 12.6 

infections. The high prevalence of Cryptosporidium is noteworthy. 

Continued diagnosis remains valuable to guide treatment and 

prevention practices. 

1008 Changes of Lymphocytes Subpopulation and Serum 

Immuno-chemistry Profiles of Colostrum Conferred Korean 

Native Calves during the First 21 th Days Postpartum 

YH. Jung 1 , TY. Hur 1 , SJ. Kang 1 , YJ. Do 1 , MS. Lee 1 , GH. Suh 2 , 

IS. Ryu 1 , DS. Son 1 , CY. Choi 1 , JT. Kim 3 

1 National Institue of Animal Science, Livestock Resources 

Development, Cheonan, Korea, South 

2 Chonnam National University, College of Veterinary Medicine, 

Gwangju, Korea, South 

3 Kangwon National University, School of Veterinary Medicine, 

Chuncheon, Kangwon, Korea, South 

This study was carried out to changes of lymphocytes subpopulation 

and serum immuno-chemistry profiles of colostrum conferred 

Korean native calves during the first 21th days postpartum. A total 

of 65 calves were used and blood samples were collected through the 

cervical vein by serum separator from added vacuum tubes on 1st 

(pre-feeding colostrum), 2 nd , 3 rd , 7 th , 14 th and 21 th day of 

postpartum. The percentages values of CD2 + , CD4 + , CD8 + , CD26 + , 

ACT2, MHC I, MHC II, WCI and B cells were determined using 

monoclonal antibodies to cell surface marker(VMRD, Pullman, 

USA) and FACScanto flow cytometer (Becton Dickinson 

Biosciences, San Jose, USA). Serum was separated from the blood 

immediately after the collection and stored in a -70°C deep-freezer 

until analysis. The proportion of MHC II cell in peripheral blood 

leukocyte (PBL) was significantly higher at 1st than 2nd 

day(p10 6 CFUs per ml), but 5 were not (10 6 CFUs per ml). Sixty-five isolates 

from seven calves were tested for antibiotic sensitivity by the agar 

disc diffusion assay. All were resistant to erythromycin, lincomycin 

and tetracycline and 54 to penicillin G and tylosin, but none was 

resistant to florfenicol. More cases will be studied and the C. 

perfringens isolates will be toxin typed. Further research is also 

needed to identify the aetiology of sudden death of bacteriologically 

negative calves. 

1011 Efficacy of Florfenicol Associated or not with Intravenous 

Fluid Therapy for Treatment of Experimental Salmonellosis 

in Neonatal Calves 

D. Silva, P. Silva, J. Fagliari 

Faculdade de Ciencias Agrarias e Veterinarias/UNESP, 

Departamento de Clinica e Cirurgia Veterinaria, Jaboticabal-SP, 

Brazil

The aim of the study was to evaluate the efficacy of florfenicol 

combined or not with IV fluid therapy for treatment of Salmonella 

dublin-infected calves. Twenty four 10 to 15-day-old Holstein calves 

were randomly allotted in four groups with six calves each: control 

(group 1); calves infected with 10 8 CFU of Salmonella Dublin orally 

and without treatment (group 2); calves infected with 10 8 CFU of S. 

Dublin orally and treated with two intramuscular injections of 20 

mg/kg of florfenicol, apart 48h (group 3); and calves infected with 

10 8 CFU of S. Dublin orally and treated with florfenicol + 60 

ml/kg/day of Ringer lactate solution IV (group 4). Animals were 

submitted to physical examination before inoculation and at every 

24h, during seven days after infection. At the same time, rectal 

swabs and blood samples were collected for S. Dublin isolation and 

for pH and blood electrolytes determination, respectively. Data were 

analyzed by use of ANOVA for repeated measures followed by F 

tests. A value of P

1015 Prevalence of Eimeria Infections in Dairy Calves from the 

state of Minas Gerais, Brazil 

EJ. Facury Filho 1 , M. Moura 1 , P. Soares 2 , PM. Ferreira 1 , 

AU. Carvlho 1 , M. Ferreira 1 

1 Escola de Veterinária da UFMG, Departamento de Clinica e 

Cirurgia Veterinaria, BELO HORIZONTE, Brazil 

2 Bayer SA, Division Animal Health, Sao Paulo, Brazil 

As Brazilian milk production growth rates rise, the importance of 

infectious diseases increase. In this scenario, coccidiosis may play a 

relevant role in calf health. The main objective of this study was to 

evaluate the prevalence of Eimeria infections in dairy calves up to 1 

year of age, in important dairy regions of the State of Minas Gerais, 

the largest dairy belt of Brazil. Nine hundred two feces samples were 

collected in 20 semi-intensive dairy farms from December 2006 to 

January 2007, from newborns to 1-year-old Holstein and Zebu crossbreed 

calves. Eimeria oocysts were found in all the farms, present in 

66% of the samples. The results show that OOPG is higher in 

younger calves up to 60-days-old (2585 oopg), but with lower 

prevalence (35%), while as age progresses, prevalence increases 

(73.18%) and OOPG decreases (324 oopg). Ten Eimeria species 

were identified, and E. zuernii, E. bovis, E. cylindrica, and E. 

ellipsoidalis were the most frequently found. E. zuernii showed 

higher prevalence in younger animals and E. bovis in animals from 6 

to 12-months-old. Eimeria infections can be considered endemic in 

the region studied, with younger calves more intensely affected with 

pathogenic species, which confirms its importance as a calf disease 

in Brazil. 

Key words: eimeriosis, coccidiosis, calves diseases, epidemiology 

1016 Prevalence and Risk Factors for Abomasal Lesions in Veal 

Calves 

C. Baehler 1 , G. Schüpbach 3 , M. Stoffel 2 , A. Steiner 1 , A. Von Rotz 2 

1 

Clinic for Ruminants, Vetsuisse Faculty, University of Bern, Bern, 


2 

Institute of Veterinary Anatomy, Vetsuisse Faculty, University of 

Bern, Bern, Switzerland 

3 

Swiss Federal Veterinary Office, Bern, Switzerland 

Over a period of 6 months, 125 calves were randomly selected at one 

large abattoir in Switzerland. 64 calves natura and 61 conventional 

were included in the study. The abomasa were classified according to 

the presence or absence of lesions in the fundic and pyloric parts. Risk 

factors for lesions of the 2 anatomical regions were analysed 

separately, because the etiology was reported to be different. 

Key words: veal calves, abomasal lesions, prevalence, risk factors 

1017 Influence of Supplemented Yeast-selenium on 

Thermometabolism in Iranian Holstein Suckling Calves 

A. Towhidi 1 , M. Ebrahimi 1 , A. Nik khah 1 , M. Sanjabi 2 

1 University of tehran, Department of Animal Science, Karaj, Iran 

2 IROST, Department of Animal Science, Tehran, Iran 

The Objectives of this study was to determine the effects of 

supplemented milk with Selenomethionine (Yeast-selenium) on 

thermo-metabolism in Holstein calves.10 Iranian Holstein suckling 

calves with 1 month of age were randomly allocated in 2 groups and 

fed unsupplemented milk (control)or supplemented milk with 0.3 

ppm Se-Met/kg in dry matter intake of milk (treated group) during 

winter. The animals were received the treatments and free choice 

starter for 2 months. The calves were weighed every week and 

blood samples were obtained weekly through the jugular vein, and 

plasma were obtained and frozen at -20 °C. Plasma concentrations 

of T3, T4 and T3-Uptake were determined using radioimmunoassay 

kits. Plasma cholesterol and glucose concentrations were 

determined by the Enzymatic, Colorimetric method. Rectal 

temperature was recorded at bleeding time. Environmental 

humidity and temperature were measured every day over the 

experiment. Data were analyzed by using Mixed Model. Treatment 

affected (p

The results obtained in sick animals were compared with results in 19 

healthy animals. The analyses in sick animals showed significantly 

lower mean serum concentrations of P, Alb (P

group showed higher serum lactate concentrations than the control 

group. 

Conclusions: The blood pH evolution suggests that their values were 

influenced by both the addition of malate and the diet fed in each stage 

(growing or finishing). In fact, supplemented animals showed more 

stables values than controls whose values fluctuated, and our data show 

that malate addition did not give any beneficial response in comparison 

with controls in relation to blood buffer bases (HCO 3- and BE). The 

observed fluctuations in blood pCO 2 did not allow us to establish a 

clear trend and, finally, in the present study serum L-lactate remained 

consistently higher in malate-supplemented animals than in controls, 

indicating that the effects of malate might be dependent on the 

characteristics of the diet. 

Key words: malate, acid-base, calves, productive parameters 

1023 Treatment of Systemic Acidosis in Calves by Administration 

of Sodium Bicarbonate with Stomach Tube 

D. Zimmermann, R. Mansfeld 

University of Munich, Clinic for Ruminants, Munich, Germany 

Objectives of study: The objective of this study was to investigate to 

what extend it is possible to treat metabolic acidosis in diarrheic calves 

by intraruminal administration of sodium bicarbonate and if the pH of 

the ruminal fluid influences the resorption of the buffer. 

Materials and Methods: 32 calves up to 4 weeks of age with 

metabolic acidosis caused by diarrhea (base excess: -3 mmol/l to -15 

mmol/l) were divided into 2 groups on the basis of their ruminal 

fluid pH. The 21 calves in the first group showed an acidemia 

without a rumen acidosis, the 11 calves in the second group showed 

both acidemia and rumen acidosis. All calves were completely 

rehydrated with a standard infusion solution before the application 

of the buffer. Thereafter, 50 g sodium bicarbonate dissolved in 500 

ml of water was given to all calves using a stomach tube. Blood 

samples were regularly taken. Blood gas values, hematocrit, D- and 

L-lactate, electrolytes, the pH of the ruminal fluid and the pH of the 

urine and its specific weight were determined. At three measurement 

times the degree of dehydration, respiration and state of health were 

examined and valued by a score system. 

Results: After oral administration of sodium bicarbonate 56.7% of the 

treated calves were cured permanently. This study showed no influence 

of the ruminal fluid pH on the resorption of sodium bicarbonate in the 

rumen. The less a calf was drinking after therapy of acidosis, the more 

probable it was that it relapsed and got an acidosis again. Obvious 

adverse effects of the intraruminal sodium bicarbonate administration 

did not occur in this study. 

Conclusions: 50 g of sodium bicarbonate seems to be an adequate 

dosis for the compensation of a low or medium metabolic acidosis. 

Nevertheless symptoms of overdosage or underdosage must be 

cautiously observed. If the calf is not able to compensate water 

balance, electrolyte balance and energy balance by sufficient 

independent intake of fluid and particularly milk, infusion therapy is 

strongly suggested. 

Key words: calf, acid-base balance, acidosis therapy, buffer, pH of the 

ruminal fluid 

1024 High Incidence of Icterus, Neurological Symptoms and 

Congenital Hepatic Fibrosis in Newborn Calves in a Dairy 

Farm in the Czech Republic 

M. Skoric 1 , P. Fictum 1 , P. Fleischer 2 , O. Langova 2 

1 University of Veterinary and Pharmaceutical Sciences, Faculty of 

Veterinary Medicine, Institute of Pathological Morphology, Brno, 


2 University of Veterinary and Pharmaceutical Sciences, Faculty of 

Veterinary Medicine, Clinic of Ruminant Diseases, Brno, Czech 

Republic 

We recorded increased incidence of liver disease in newborn calves 

in a farm with 400 Siemental cows. These cows were fed silaged 

milled grains of barley and maize of low quality (mouldy maize) 

from autumn 2006. Poor quality clover haylage should not have been 

used for feeding, however, it partially was. There was some increase 

in the somatic cell count in milk and increased occurrence of 

mastitis. At the end of the year one third of calves (fed by natural 

milk) diseased, mostly at the age of 10 days, and 50% of them died 

within next 5 days. Remarkable clinical signs were lethargy, 


inappetence, eventually mild diarrhoea, seizures of convulsions, and 

after several days icterus (but without fever). The main biochemical 

changes in the blood of 12 calves were extremely high levels of total 

bilirubin (21-344; x=111 µmol/l) and AST (3.9-33.8; x=15.0 µkat/l). 

Further findings (lower levels of Ig, total protein, GMT and vitamin 

A) showed in one half of the calves having problems with colostrum 

nutrition, in spite of good technology. In case of inappetence the 

milk was administred by esophageal feeder (till 3-4 days). This 

procedure probably led to rumen acidosis. One shortly hospitalized 

calf had metabolic acidosis, which was caused by severe rumen 

acidosis (pH 4.37). Two animals were subjected to necropsy, in 

another case only liver was examined. We observed pathological 

changes in liver, which were represented by mild hyperaemia and 

structure of liver parenchyma on the cut surface resembling nutmeg. 

Histopathological examination of liver from all three calves showed 

the presence of small islets of hepatocytes separated from each other 

by fibrous tissue with presence of proliferating and abnormally 

branching bile ducts. The hepatocytes showed signs of vacuolar 

degeneration, foci of focal coagulative and haemorrhagic necrosis, 

cholestasis and hyperaemia were present in parenchyma. There was 

round cell, predominantly lymphocytic, infiltrate in the proliferating 

fibrous tissue. We detected marked microscopical lesions morphologically 

corresponding with congenital hepatic fibrosis in examinated 

calves. Examinations for leptospirosis, listeriosis, selenium 

overdosing and nutritional muscle dystrophy were negative. The 

cause of the disease could be chronic intoxication of cows by 

substances diffusing to the fetus and into the milk, e.g. aflatoxins or 

products of protein degradation. 

Supported by the grant MSM6215712403 

Key words: cattle, intoxication, liver, necropsy. 

1025 Prophylactic Measures and Epidemiology of New born Calf 

Diseases in Korea 

IS. Ryu 1 , YJ. Do 2 UH. Kim 1 , TY. Hur 2 , SJ. Kang 2 , CY. Choi 2 , 

YH. Jung 2 , DS. Son 2 , JS. Ko 2 , HR. Han 3 

1 

National Institute of Animal Science, Technology Application 

Division, Suwon, Korea, South 

2 

National Institute of Animal Science, Animal Resources 

Development, Cheonan, Korea, South 

3 

Seoul National University, College of Veterinary Medicine, Seoul, 

Korea, South 

To set up prophylactic measures for new born calf diseases, we 

analyzed the incident rate and the causes using 1118 new born 

calves of 23 farms by epidemiology investigation. As investigation 

results of the cases of diseases in 597 new born calves, the cases of 

diarrhea, pneumonia and septicemia were 318, 49 and 19 cases 

respectively and then diarrhea and broncheitic pneumonia 3 

cases, downer cow was 2 cases and weakness, dermatitis and 

indigestion was a case. In the species, 153 cases were in 229 

newborn calves of Han-woo and 204 cases were in 376 newborn 

calves of Holstein. Therefore the incidence in Holstein was 2times 

of in Han-woo. As a result of the analysis of death causes in 85 

newborn calves, diarrhea, septicemia, pneumonia, BVD, weakness, 

hoove and arthritis were 23, 8, 7, 6, 4, 3 and 1 respectively and 

Akabane disease, indigestion and downer cow were respectively 2. 

As the incidence in Han-woo, below 20kg was 25%, 21~25kg was 

7.9%, 26~30kg was 4.3%, above 31kg was 0%. As a result of blood 

and feces analysis, E. coli 9, Rota virosis 7, Coccidiosis 6, 

Salmonellosis 6, E. coli Coccidiosis 3, E. coli and Rota virosis 3, 

Coccidiosis and Clostridium 3, Clostridium 2, enterotoxemia 1 and 

etc. To prevent WCS (Weak calf syndrome) in pregnant cow, 

6(29%) calves died after Vitamin A, D, E (Vigantol-E ® , Bayer) 5ml 

IM, on the other hand 15 (71%) calves died before that time. As a 

result of the blood chemistry analysis, Ca, Mg, CRE, GGT, GOT, 

TP, BUN, GLU and CHO were in normal range, and ALB, LDH, 

PHO, TBI and TRI were in over range. In these results, death causes 

of new born calves in domestic cow in Korea were mainly diarrhea 

and respiratory disease by bacteria and virus. So it is thought that 

Vigantol-E ® injection before parturition and application 

vaccination program about diarrhea and respiratory disease can be 

help to prevent disease of new born calves and WCS. 

Key words: calf, WCS 

1026 Serum and Acute Phase Proteins in Calves with Respiratory 

Syndrome

Cs. Tothova, O. Nagy, H. Seidel, J. Konvicna, G. Kovac 



Acute phase proteins are a class of proteins that are synthetised 

predominantly in the liver in response to inflammation, tissue 

damage, transport, stress, starvation or exhaustion of organism. The 

increased production of these proteins is more typical for acute 

inflammatory processes, than for chronic conditions. The objective 

of this study was to evaluate the influence of chronic respiratory 

diseases in calves on the concentrations of selected acute phase 

proteins: haptoglobin (Hp) and serum amyloid A (SAA), as well as 

on the concentrations of some variables of protein metabolism: total 

proteins, albumin, urea, creatinine, and total immuno-globulins in 

blood serum. The results obtained in calves affected by chronic 

respiratory diseases we compared with values determined in 

clinically healthy calves. The evaluation was performed in 11 calves 

with chronic respiratory disease and in 11 clinically healthy calves 

of a low-land black spotted breed and its crossbreeds at the age of 4- 

6 months. Hp and SAA were determined by method of enzyme 

linked immunosorbent assay. The determinations of total proteins, 

albumin, urea, and creatinine concentrations were performed on 

automatic biochemical analyser. Total immunoglobulins were 

assessed by spectro-photometric turbidimetric method (zincsulphate 

test). In calves with chronic respiratory disease we recorded 

markedly higher average concentration of Hp than in healthy calves. 

By evaluation of SAA concentrations in the group of calves with 

respiratory problems we observed no significant differences 

compared with the group of clinically healthy calves. The average 

concentration of albumin was significantly lower in diseased 

(P

thymus in calves and heifers with chronic pneumonia for monitoring 

thymus objectively in diseased individuals. 

Materials and methods: The cross-section of cervical thymus in 46 

clinically healthy calves and heifers (normal group) and 8 calves 

with chronic pneumonia (chronic group) was observed by 

ultrasonograph with 3.5 MHz and 5.0 MHz transducer. The weight 

of the thymus of the chronic group was measured and compared with 

the cross section of the cervical thymus. They were 1-100 weeks of 

age in normal group and 5-60 weeks of age in chronic group. 

Results: The cross section of cervical thymuses of normal group 

were increased from born to 31 weeks and made plateau to 45 weeks 

and space decreased to 100 weeks. The mean cross sections of 

cervical thymus at 1-5 weeks and 31-35 weeks were 5±1.4cm 2 and 

22.9±0.4cm 2 , respectively, and were 4-fold increased compared to 

that of birth, and then it decreased to 3.7±0.5 cm 2 at 96-100 weeks 

after birth. The mean cross section of the cervical thymus in chronic 

group at 1-5, 31-35 and 41-46 weeks were 3.1(±1.9) cm 2 , 6.1 cm2, 

and 3.7 cm 2 , respectively and these values were significantly 

lowered, compared to those of normal group. The echogenic of the 

cross section of cervical thymus in normal group at±2 weeks of age, 

37 weeks of age and 87 weeks of age were hypo echoic, moderate 

hyper echoic and hyper echoic. A positive correlation (r=0.74) was 

found between the cross section of thymus and their weights in 

chronic group. The body weight was decreased and prognosis was 

bad in the chronic group. 

Conclusion: The observation of thymus by ultrasonograph is an 

effective tool for estimating the size of thymus in Holstein calves and 

heifers from birth to 100 weeks of age. The cases of chronic pneumonia 

of small size thymus are bad prognosis. The results of this study prove 

to be useful and applicable to monitor the growth of thymus in Holstein 

calves and heifers. 

1030 Probiotics in Prevention of Diarrhea in Calves 

J. Prasek 1 , J. Illek 2 , D. Kumprechtova 2 

1 Veterinary and Pharmaceutical University Brno, Microbiology and 

Immunology, Brno, Czech Republic 



Objective: The aim was to investigate the effects of dietary 

supplementation with LACTIFERM L5 (Enterococcus faecium M74 

(NCIMB 11181) on prevention of diarrhea, health, performance, and 

blood chemistry parameters in dairy calves under field conditions. 

Material and methods: 24 Holstein calves (8 males and 16 

females). Experimental treatments: 1. Control (C), n = 12, 2. 

LACTIFERM (L), n = 12. First 5 days of life the calves received 

colostrum (2 L 2x/day). The L calves received 5g probiotic/calf/day, 

7.2 x 10 9 CFU/1g. Calf starter and water were provided ad libitum. 

Starter consumption was measured daily. The trial lasted 50 days, 

scours occurence was recorded. The calves were weiged on days 1, 

21, 42 and 50, average daily weight gain (ADWG) was calculated. 

Blood samples were collected on days 1 and 50 and 3 times in 

between (5 collections), and analysed for total protein (TP), albumin, 

immunoglobulins (Ig), urea and Ca, Mg, Cu, Zn. 

Results: ADWG for the whole experimental period was 0.61 kg/day 

and 0.58 kg/day in L and C, respectively. The difference was not 

significant. Calf starter consumption per kg weight gain was 

significantly (P

cases of diarrhea recorded. The cumulative incidence rate of diarrhea 

was significantly (p

concentrations of calves with umbilical hernia were determined by 

means of sodium dodecyl sulphate-polyacrylamide gel 

electrophoresis and were compared with time 0. 

Results: Five plasma and peritoneal fluid proteins (ceruloplasmin, 

transferin, albumin, haptoglobin, and acid glycoprotein) were 

increased in calves with umbilical hernia after surgery, compared 

with baseline values. Peritoneal fluid findings: ceruloplasmin 

(201.67 to 530.33%) and transferin (117.28 to 251.85%) showed the 

highest percentage increase in concentration from day 1 to 15. The 

highest percentage increase in concentration of haptoglobin (48.67 

to 400%) and albumin (66,41 to 131,83%) were observed from day 1 

to 7 and were lower than the other two previously mentioned. Acid 

glycoprotein showed the highest percentage increase in 

concentration on day 3 (355.4%). Plasma protein concentrations: 

Haptoglobin was the protein with the highest percentage increase in 

concentration (88.39 to 366.6%) from day 1 to 5. These values were 

higher than those found in the peritoneal fluid during the same 

moments. 

Conclusions: Results indicate that acute phase proteins increase more 

rapidly after the onset of inflammation in the peritoneal fluid than in 

plasma, except for haptoglobin. Because of this, it may be possible to 

use measurements of these proteins to monitor the progression of the 

inflammatory response in the abdominal cavity after surgery. 

Supported by grants from FAPESP (07/51389-4). Authors thank Mr. 

Paulo César da Silva for his invaluable help in the acute phase protein 

quantitations. 

Key words: umbilical hernia, calf, acute phase proteins 

1037 In vitro Evaluation of Curd Formation after Reconstitution 

of Neonatal Calf Oral Rehydration Therapy Solutions in 

Whole Cow’s Milk 

G. Nappert 1 , P. Domps 2 

1 

Lachute Veterinary Hospital, Veterinary Consultancy and Research, 

Lachute, Canada 

2 

CEVA santé Animale, Livestock Business Unit, Libourne, France 

Objective: To compare the in vitro effects on whole cow’s milk curd 

formation of 40 neonatal calf oral rehydration therapy (ORT) solutions 

commercially available in Europe and in Canada. 

Materials and Methods: Homogenized raw milk from a 

commercial dairy farm was used and kept at 30 °C on a magnetic 

stirring module with an electronic contact thermometer. The ORT 

solutions were reconstituted in triplicate in 50 mL of whole cow’s 

milk according to the manufacturer’s directions. The pH and the 

osmolarity of each test solution were measured after 

reconstitution. In vitro milk clotting was performed in triplicate, 

using rennin as the clotting agent. The rennin, extracted from calf 

abomasum, was diluted with double-distilled water to a 

concentration of 12.5 g/L with a proximal activity of 50 U/mg 

protein. The rennin solution (0,1 mL) was added to each test solution 

(50 mL). Measurement of clotting time was determined by a sudden 

variation in the conductivity of the solution within a 10-minute 

period. Objective assessment of the weight of the coagulum was 

measured with a desiccator scale. 

Results: After reconstitution, only four products had osmolarities 

between 400-600 mmol/L. ORT solutions containing large amounts of 

bicarbonate, and/or citrate (>40 mmol/L), presented a significant 

alkaline pH after reconstitution and had negative effects on curd 

formation. Glucose-rich ORT solutions also had significant negative 

effects on milk clotting time. Isotonic oral electrolyte solutions that 

contained mainly acetate and propionate did not interfere with milk 

clotting time and curd formation. 

Conclusions: In accordance with previous studies, the feeding of 

diarrhoeic calves with whole cow’s milk is recommended in 

combination with isotonic metabolisable base ORT solutions. It has 

already been documented that suckling bicarbonate/citrate-rich ORT 

solutions result in sustained abomasal alkalisation. For diarrheic 

calves, this may reduce the digestibility of milk. 

Key words: calves, diarrhea, rehydration, milk, electrolytes 

1038 Study of the Efficacy of Hiprabovis Pneumos in front of a 

Challenge Infection with Mannheimia haemolytica in Young 

Calves 

A. Foix, S. Casademunt, R. March 

Laboratorios Hipra, S.A., Amer (Girona), Spain 


Objective: To demonstrate the protection conferred to calves infected 

with M. haemolytica by a vaccine against Mannheimia haemolytica 

and Histophilus somni. 

Materials And Methods: The vaccine Hiprabovis Pneumos (Laboratorios 

Hipra, S.A.) of M. haemolytica A1 leukotoxoid (450 IEU 

per dose) and H. somni (1x10 10 bact/dose) was tested (group A, 6 

calves), and compared to non-vaccinated (placebo)+ infected (group 

B, 6 calves) and to non-vaccinated + non-infected (group C, 2 calves 

as sentinels). The vaccine and placebo (PBS) were administered 

twice, at the dose of 2 ml, separated 21 days. Groups A and B were 

experimentally infected in day 42 intratracheally with 1x10 10 cfu of 

M. haemolytica in 10 ml per calf. Efficacy was evaluated by 

studying clinical signs (scoring system based on rectal temperature, 

apathy, anorexia, nasal discharge, lung auscultation, conjunctivitis, 

dyspnoea and cough), macro and microscopic lung lesions and lesion 

size. No other treatments were given during the study to these 

animals of 100-150 kg and of 2 months of age. Presence of 

antibodies to M. haemolytica leukotoxoid was an exclusion factor.at 

beginning of the study. Mean and standard deviation were calculated 

for rectal temperatures, clinical signs and lung lesions score at 

different days post-vaccination for each group of calves. A t-test and 

a Mann-Whitney test were used for continuous and discontinuous 

variables respectively (P=0.05). Animals of group C were not 

included in the statistical analysis. 

Results: Clinical differences between infected (A, B) and noninfected 

(C) proves the success of the method of infection. The 

clinical signs score (0 to 3) showed differences between groups A 

and B at day 1 (p=0.06), day 4 (p=0.02) and day 7 (p=0.007). After 

infection 2 calves of group B died (days 1 and 6). Mean rectal 

temperature was different for groups A and B at day 1 post-challenge 

(p=0.02). Percentage of pneumonic pulmonary tissue was: A = 

18.1%; B = 51.2% (A-B, p=0.01); C = 13.9%, and this means a 

reduction of 64.6% for the vaccinated group. The severity of 

pulmonary lesions score (0 to 5) was: A = 1.8; B = 3.8 (A-B 

p=0.009); C = 1.5, and this means a reduction of 52.6% for the 

vaccinated group. 

Conclusions: Hiprabovis Pneumos proved to be effective in front of an 

experimental infection with M. haemolytica as it did reduce the clinical 

symptomatology, rectal temperature, percentage and severity of 

pneumonic tissue. 

1039 Neutrophil Oxidative Burst in Nelore and Limousin Calves 

in the First Four Months of Life 

M. Costa, K. Flaiban, R. Dognani, E. Vettorato, M. Balarin, J. Lisbôa 

Universidade Estadual de Londrina, Londrina, Brazil 

Objectives: To study the oxidative burst of neutrophils in Nelore and 

Limousin calves in Brazil during the first four months of life. 

Materials and Methods: 90 healthy calves (45 Nelore; 45 

Limousin) were randomly selected from two farms in Parana State, 

Brazil. Blood samples were collected at 24 to 36 hours, 15, 30, 60, 

90, and 120 days of life. Neutrophil oxidative burst was determined 

by the non-stimulated (NS-NBT) and stimulated (S-NBT) nitroblue 

tetrazolium test. The variation of positive neutrophils during the 

various ages was analysed by the Krushkal-Wallis statistical test. 

Comparisons between the breeds at different ages were done using 

the Mann-Whitney statistical test. 

Results: The percentages of reactive neutrophils to the NS-NBT test 

were 2.5, 2.0, 2.0, 3.0, 3.0 and 2.0 for Nelore and 2.0, 2.75, 4.0, 4.0, 5.0 

and 5.0 for Limousin calves on the above cited ages. For the S-NBT, 

the values of 6.0, 6.0, 6.5, 8.0, 10.5 and 6.0 for Nelore and 6.0, 8.0, 8.5, 

9.5, 15.5 and 15.0 for Limousin calves were obtained. 

Conclusions: The neutrophil oxidative burst was less effective in 

newborn calves increasing with age in both breeds. There was a 

significant increase in the stimulated test by 90 days of age. 

However, this activity decreased at 120 days in the Nelore calves. 

Such an increase can be due to the immune response against 

hemoparasites, which are normally encountered at this age in calves 

kept under range conditions in tropical countries. Statistical 

differences among the two breeds were observed at 30, 90 and 120 

days in the NS-NBT and at 15, 30, 90 and 120 days in the S-NBT. 

Although, Limousin calves had a higher neutrophil activity, this 

difference had no practical importance in the health of the studied 

calves, probably because of the low challenge observed in animals 

kept on pasture and the good management practices on both farms. 

Younger calves can be considered to be more susceptible to disease

ecause of their lower neutrophil activity. However, other defence 

mechanisms, as passive immunity and higher neutrophil count 

should be considered. The present results suggest that this increase is 

physiologic in cattle. Further studies are necessary to investigate 

neutrophil oxidative burst in sick calves. 

Key words: zebu, taurine, beef calf, NBT 

1040 A Comparison of the Efficacy of Baytril 100 and Bio-Mycin 

200 for the Treatment of Bovine Respiratory Disease in 

Feedlot Cattle 

S. Ives, J. Sears 

Bayer Health Care, USA, United States 

A study was conducted to evaluate the relative efficacy of Baytril 

100 (enrofloxacin) to Bio-Mycin 200 (oxytetracycline) as the initial 

therapy in feedlot cattle considered to have a moderate risk for 

experiencing respiratory disease. Steers and heifers were procured 

from several order buyers, transported to the study site and subjected 

to a common processing regimen, which did not include 

administration of a metaphylactic antimicrobial. Individual animals 

that met enrollment criteria, were treated in pairs, so that one animal 

received Baytril 100 (5cc/100 lb body weight) and the other Bio- 

Mycin 200 (4.5 cc/100 lb body weight). Animals were penned 

according to gender and enrollment treatment, so that at the 

completion of the enrollment period, 473 treatment pair groups (946 

hd) had been placed on study. Cattle were maintained in 14 

enrollment pens (seven per treatment) until cattle were reimplanted, 

at which point, pens were combined according to pen size, 

enrollment treatment and gender, so that three distinct feeding 

groups per treatment were formed. Cattle were kept in their feeding 

groups until harvest, at which point, final group weights and carcass 

data were obtained. Case fatality rates for animals treated with 

Baytril 100 were reduced as compared to animals treated with Biomycin 

200 (26.7% vs. 33.3%, respectively; P=0.02). Differences in 

carcass parameters were not detected between groups, as cattle that 

survived to harvest tended to peform similarly. All cattle enrolled in 

this study had a negative financial return, however, the animals 

treated intially with Baytril 100 had an economic advantage of 

$46.22/head as compared to those treated initially with Bio-mycin 

200. The respiratory case fatality rates experienced were 

substantially higher than expected for these cattle. The primary 

reason was protracted adverse winter weather events occurred during 

the conduct of the study which superimposed additional 

physiological stress at critical points in the receiving period and 

adversely affected these cattle for the remainder of their time in the 

feedlot. The results of this study suggest that Baytril 100 may be 

more cost effective than a lower cost therapy such as Bio-mycin 200 

even in cattle thought to be at a more moderate level of risk for 

experiencing BRD. 

1041 The Dynamic of Fetal Hemoglobin in Calves of Different 

Breeds from Birth to the 180 th Day of Life (age): Influence of 

Breeds 

M. Caribe Ayres 1 , E. Birgel 2 , AC. Macedo 1 , R. Viana 3 , P. Soares 4 , 

E. Birgel Junior 2 

1 Escola de Medicina Veterinária UFBA, Departamento de Patologia 

e Clínicas, Salvador, Brazil 

2 Faculdade de Medicina Veterinária e Zootecnia, Departamento de 

Patologia e Clínicas, Sao Paulo, Brazil 

3 Universidade Federal Rural da Amazônia, Departamento de 

Medicina Veterinaria, Belém, Brazil 

4 Universidade Federla Rural de Pernambuco, Departamento de 

Medicina Veterinaria, Recife, Brazil 

Fetal and adult hemoglobin of various species behave differently in 

their resistance to acids and alkalis, oxygen dissociation curves, 

electrophoretic mobility, ultraviolet absorption and in amino acid 

composition. Observations to determine the status of postnatal 

duration of fetal hemoglobin in the bloodstream have been report in 

humans. The hereditary persistence of fetal hemoglobin (HPFH) is a 

condition characterized by the continue expression of the fetal 

globins in adulthood and this disorders is included as a 

hemoglobinopathies. This research was the undertaken to determine 

how long the fetal hemoglobin of calves persisted postnatally, 

evaluating the influence of breed, in cattle raised in Brazil. For this 

study 38 health calves of different breeds (10 Holstein, 10 Jersey, 10 

Zebu-Holstein crossbred and eight Gir) were used to follow the 

dynamics of the fetal hemoglobin and adult hemoglobin distribution 

and the erythrogram changes. To evaluate these variable periodic 

blood sampling were carried out with EDTA, from birth to the 180th 

day of life. To quantify the fetal hemoglobin and adult hemoglobin a 

densitometer method was used and this determination was preceded 

by electrophoresis separation of the fetal hemoglobin and the adult 

hemoglobin. Red blood cells were washed three times by addition of 

0.9% NaCl solution, centrifugation, and removal the supernatant. 

The washed cells were then hemolyzed with distilled water and this 

solution was frozen at 4 °C and again they were washed by addition 

of chloroform and stroma was removed from the supernatant. The 

hemoglobin solution was stored at – 80 °C. Analyses were made on 

a BioSystem Electrophoreis and the hemoglobin solution was 

banded upon on a acetate cellulose and the strips were colored in 

Amido Black solution. Statistical methods were used and after 

radical transformation of the hemoglobin percents obtained for the 

different groups. At the fist day of life most hemoglobin (87%) was 

fetal. At the 150 th day the Holstein, Jersey and crossed calves had 

only adult hemoglobin while the fetal hemoglobin remained in the 

bloodstream if the Gir calves up to 165 th day. The average of the 

fetal hemoglobin percents in the birth day was highest in the 

Holstein calves. The physiology persistence in the bloodstream in 

cattle was influenced by the breed. 

Key words: hematology, genetics, fetal hemoglobin, calves, 

hemoglobinophaties 

1042 Clinical Classification of Diarrhoeas in Calves 

P. Cano-Celada, M. Mancera 

Universidad Nacional Autónoma de México,Facultad de Medicina 

Veterinaria y Zootecnia, Producción Animal:Rumiantes, Mexico, DF., 

Mexico 

In this paper we will try to show the importance of the clinical 

classification of diarrhoeas, by describing the specific clinical signs, 

depending in how long the calf has been sick and the relation with his 

dehydration rate. Mechanical diarrhoea with a 0 to 4% non apparent 

dehydration, Grade I the new born calf needs 8 to 10% of his live 

weigth of milk or calostrum intake every 24 hours, if we administer 

more milk we provoke a mechanical diarrhoea, because of the 

saturation of nutrient absorption capacity, this provokes an osmotic 

change. Acute 5 to 7% GII present diarrhea to 5 to 7 days, with 

dehydration. Chronic infectious diarrhoea with 7 to 9% dehydration 

rate, G III: In this case diarrhoea persists for 7 to 10 days, with 

dehydration, haemoconcentration, vasoconstriction, metabolic 

acidosis, respiratory alkalosis, bacteraemia, and toxemia with a 40 °C 

fever, tachycardia polipnea, anorexia, depression, very fluid stinky 

diarrhoea, adipsia, dry snout, submerged eyes, pale and dry mucosa, the 

calf is skinny, and the skin is dry and rough and when you pinch it, 

elasticity delays 9 to 15 seconds to return. Severe Diarrhoea and 

dehydration from 10 to 12%, G IV. In this cases diarrhoea and 

dehydration have been present for more than 15 days, it can be synergic 

activity among viruses, bacterias and parasites such as E. coli and 

Rotaviruses, clinical signs became worst in a progressive way until 

they provoke depression, weakness, total anorexia, fever above 41 °C 

because of sepsis and toxemia, tachycardia with arriythmia, polipnea, 

pale mucosa, stinky diarrhoea with blood strips or melaena, very 

concentrated urine, sphincter weakness, the calf lies down and skin 

elasticity when you pinch it returns in 24 to 45 seconds. Serious 

diarrhoea with an over 12% dehydration, G V: This a very serius 

diarrhoea skin remains together when pinched, we can see severe 

dehydration, electrolyte unbalance, immune depression, 

metabolic acidosis, respiratory alkalosis, ketosis, hypothermia, uremia, 

sepsis, toxemia causing meningitis, shock, seizures and dead, nervous 

signs indicate the gravity of the dehydration. Overacute diarrhoea. In 

this cases, very susceptible calves die in 72 hours because of extreme 

pathogenic aetiologycal agents such as superbacterias, in other cases 

the case can be E. coli. 

1043 Diarrhoea Treatment and Hydrotherapy in Calves 

P. Cano-Celada, A. Olguin y bernal, M. Quiroz-martinez 


Veterinaria y Zootecnia, Producción Animal:Rumiantes, Mexico, DF., 

Mexico 


The goal of this paper is to expose the importance of treatment and 

hydrotherapy Mechanical diarrhoea with 0 to 4% dehydration: Stop 

administering milk, correction of the exceeded amount, 

administration of serum orally. Acute diarrhoea with 5 to 7% 

dehydration: substitute milk administration for oral electrolytic 

serum we recommend to start treatment and hydrotherapy as soon as 

possible to avoid serious electrolytic unbalanced, acidosis and 

chronic infection. 10 % of live weight must be administered in oral 

serums. Apply antibiotic treatment orally and parenterally with the 

same substance to avoid antagonisms, intoxication or allergies. 

Parenteral treatment penicillin 22.000 I.U./kg/24 hrs/7 days I.M., or 

oxitetraciclins 10 mg/kg/24 hrs7 days I.M., I.V. Chronic diarrhoea 

with a 7 to 9 % dehydration. Administration of oral and for 

parenteral hydrotherapy use the following formula: body weight 

multiplied by the dehydration rate, divided over one hundred, the 

result is the hydrotherapy volume that must be administered every 24 

hours. In cases of metabolic acidosis with hiponatremia the sodium 

hypertonic alkalinizing solutions are indicated. Administer 

Gentamicin 3 mg/Kg/24 hrs/5 days, I.M., I.V. via, or sulphatrimetoprim 

25 mg kilo/24 hrs/8 days, I.M., I.V. Piroxicam 

administration 1 ml/25 kg/24 hrs deep I.M. mix of 5 ml of sulfatrimetoprim 

+ 5ml of bismute salicilate + 5 ml of caolin and pectin 8 

hours. To avoid Coccidias you should administer toltrazuril. For 

appetite stimulation administer vitamins, brotizolam or CC51 with 

Ciproheptadine. To stimulate the immune system use caseinates. 

Severe diarrhoea with dehydration from 10 to 12% with 

hydrotherapy you get blood volume back to normal levels and that 

starts again the real and hepatic function and the acid-base and 

electrolytic balance, if miction starts in the 60 minutes after the 

administration of hydrotherapy and stimuli response it means that it 

is working and we will avoid the risk of uraemia. Administer 

Ceftiofur 1 mg/Kg/24 hrs and flunixin meglumin 2.2 mg/kg/24 hrs. 

Serious diarrheoa with an over 12% of dehydration In this cases the 

calf shows nervous signs so we need to avoid seizures administering 

Xilacine 0.2 mg kilo I.V. and Hartmann serum also via I.V. via, 

added with 5 grs of sodium bicarbonate to fight acidosis. Overacute 

diarrhoea. You must administer very quickly the treatment, 

enrofloxacin 2.5 mg/kg/24hrs/ 5 days I.M., hydrotherapy must be 

administered by I.V. via 

1044 Bronchoalveolar Lavage in Calves Naturally Affected to 

Respiratory Disease: Cellular, Microbiological and 

Serological Variables 

MR. Mokhber Dezfouli 1 , H. Seifi 2 , A. Araghi 1 , P. Tajik 1 , 

S. Lotfollahzadeh 1 

1 Faculty of Veterinary Medicine, University of Tehran, Clinical 



Clinical Sciences, Mashhad, Iran 

In 1979, Wilkie and Markham introduced bronchalveolar lavage 

(BAL) in live cattle as a safe and repeatable research tool which has 

subsequently been utilized in numerous other studies for investigation 

of cellular and non-cellular immune events in the lower respiratory 

tract of cattle. The objective of this study was to compare various 

components of BAL of healthy calves with those affected with 

respiratory tract disease. Samples obtained by bronchoalveolar lavage 

(BAL) were used to evaluate pulmonary cytology, microbiology and 

serology in twenty calves with respiratory disease and eight healthy 

calves as control. Calves were sampled using BAL on the same day that 

they were designated as either cases or control. To evaluate the changes 

in microbial flora, BAL fluid were cultured on selective media for 

special bacterial and fungal pathogens of the respiratory system. There 

was no significant difference of bacteriological and fungal culture and 

serological results between cases and control calves. There was no 

significant difference of BAL variables on case and control groups. Our 

findings suggest that the elevation of neutrophils in BAL above 10% of 

the total cells may indicate pulmonary disease. In addition it appears 

that the isolation of microorganisms cultured from BAL, os not always 

consistent with the identification of causal agent of the disease. 

Key words: bronchoalveolar lavage, pulmonary disease, blood, calves 

1045 The Impact of Dairy Cows Management during the Dry 

Period on their Offspring’s Performances in Morocco 

H. Guemoune, A. Ilham, L. Maach, S. Alali 




The study aimed to evaluate the impact of management of dairy 

cows during the dry period on growth and health performances of the 

calves and also to assess the growth performance of heifers raised in 

a common rearing unit in keeping with their originated herds. The 

results obtained showed that calves performances depended on the 

management conditions applied to these animals between their birth 

and their delivery age to the rearing unit and also on some factors 

related to the cow during the dry period. Thus, the average daily gain 

(ADG) of the calves that reached 0.69 kg was influenced by the 

mother cow’s body condition score during the dry period and its 

general health status as well as by the total plasmatic protein (TPP) 

of the calves and the delivery age to the common rearing unit 

(p

norovirus may explain some of the discrepancies in detection rates 

from previous studies and shed light on some epidemiological 

aspects of bovine norovirus infections that have until now been 

elusive. 

Key words: Bovine norovirus, SYBR Green 1-based real-time RT- 

PCR, diagnostics, genotype prediction 

1048 Congenital Anophthalmia – Case Report 

K. Madureira, V. Gomes, M. Ramalho 

Faculdades Anhanguera, Medicina Veterinária, Leme, Brazil 

On October 10th, 2006, a 3-day old, Nellore bovine with white hair 

was admitted into the Department of Large Animal Clinical Practice 

and Surgery, at the Veterinary Hospital - Centro Universitário 

Anhanguera/ UNIFIAN - Leme. The animal came from a farm 

located in the countryside of the state of Sao Paulo, where extensive 

breeding was adopted. When admitted in the Hospital, it was 

observed that the animal lacked an eyeball and the tail. At the 

clinical examination, it was observed that the animal was severely 

prostrated, because it had not ingested any collostrum. Clinical 

examination also assessed heart rate (124 beats per minute), 

respiratory rate (52 movements per minute) and temperature (37.0 

°C). the animal was thin, its apparent mucous membranes were 

slightly pale, and clinical examination of the lymphnodes did not 

show any abnormalities. The animal showed an altered position of 

the anus, which was almost horizontal, and caused the contamination 

of the vulva with faces. The absence of the eyeball was assessed by 

palpation, once the animal showed eyelashes and palpebral reflexes. 

The animal was treated by fluid therapy, once it was dehydrated and 

hypoglycaemic. However, it died on the next day, and it was 

necropsied. It was observed that the animal lacked the eyeball, and 

the site was filled with connective tissue. The orbit was normal. The 

animal also showed cleft palate and absence of tail. Anus, rectum, 

vagina and uterus were present and in normal positions. Based on the 

findings of the physical examination and necroscopic examination, it 

was diagnosed that the animal showed congenital anophthalmia, tail 

agenesis and cleft palate. 

Key words: bovine, anophthalmia, agenesis 

1049 Botulism in Dairy Cattle Herd with Atypical Course of 

Disease 

K. Lutnicki, J. Marczuk, L. Kurek 

Faculty of Veterinary Medicine in Lublin, Department of Internal 

Medicine of Farm Animals and Horses, Lublin 

Neurotoxins produced by Clostridium botulinum cause botulism, a 

fatal disease of humans and animals, among which the ruminants and 

horses are most sensitive. Botulism can arise from preformed toxin, 

wound infection or intestinal toxic-infection. Among the great 

amount of exotoxins which are produced by Cl. botulinum only the 

B, C and D types are responsible for cattle morbidity. Botulinum 

toxins in cattle are very difficult to identify because this origin may 

be sensitive to levels below the detection, so in some countries cattle 

is routinely vaccinated against type C and D. Acute botulinum 

intoxication was observed in Holstein-Frezian dairy cow herd of 80 

adult cows, 30 heifers (10-12 months) and 15 calves, which were fed 

a TMR that consisted of haylage, maize silage, green forage. 

Material and Methods: The group containing 15 heifers and 4 dry 

cows (15.2% of whole herd) showed botulism signs, confirmed by 

serologic tests; no calves or cows in lactation were affected. Treatment 

was ineffective. There were haematological and biochemical analyses 

performed. 

Results: The sick animals exhibited only the increase in 

polymorphonuclear granulocytes concentration (mean 58%), which 

correlated with increased plasma AST (mean 311.9 U/l), CPK (mean 

2204.2 U/l) and total bilirubin (mean 8.54 umol/l) concentrations. 

Estimated plasma ion levels (Ca, P, Mg, Na, K) were normal, no 

significant changes were observed in comparison to healthy animals. 

There were high Cu (29.02umol/l) ion levels and low Fe (10.9umol/l) 

concentrations in affected dry dairy cows. 

Conclusions: There is a great body of evidence that lactating dairy 

cows are more resistant and did not develop clinical signs of 

botulism as easily as the other cattle. Despite the well expressed 

typical clinical signs of botulism in affected animals, estimated ions 

concentration of both calcium and sodium in plasma were not 

changed during the first period of illness in comparison to healthy 

ones. Obtained results and bibliographic data indicated that liver 

dysfunction can play the role in pathogenesis of cattle botulism. 

1050 Monitoring Immunoglobulin Status of Newborn Calves 

Using Different Immunoassays 

K. Szabó-Ari 1 , Cs. Bajcsy 1 , I. Kacskovics 2 , I. Mádl 3 , L. Takács 3 , 

O. Szenci 1 


Large Animals, Üllô, Hungary 

2 Eötvös Loránd University, Faculty of Science, Department of 

Physiology, Budapest, Hungary 

3 Agroprodukt Co., Pápa, Hungary 

Ingestion and absorption of colostral immunoglobulins (mainly 

IgG1) in newborn calves are essential for reaching an appropriate 

immune status during the first weeks of life. Failure of passive 

transfer (FPT) occurs when ingestion and absorption of colostral 

IgG1 are inadequate and the serum IgG level is below 10.0 g/L.A 

number of assays has been used to measure IgG concentration in 

colostrum and serum of newborn calves. Colostral IgG 

concentrations can be measured by different methods, such as by 

using a colostrometer, a refractometer, a lateral-flow immunoassay, 

a radial immunodiffusion or an ELISA. The following methods can 

be used for the determination of serum IgG levels in calves: sodiumsulphite 

precipitation test, zinc-sulphate turbidity test, 

glutaraldehyde coagulation test, refractometry, lateral-flow 

immunoassay, latex agglutination test, simple immunoassay, radial 

immunodiffusion, electrophoresis and ELISA. In a field trial, the 

IgG concentrations of the colostrum and of the serum from newborn 

calves were measured using lateral-flow immunoassay kits (MBC 

Midland BioProducts Co, Boone, Iowa, USA). 93.8 % of the 

examined 81 colostrum samples, withdrawn from Holstein-Friesian 

dairy cows immediately after parturition, contained adequate IgG 

concentrations (>50 mg/ml), while 6.2 % had inadequate levels. On 

the other hand, only 69.1 % of the 81 one-day-old newborn calves 

showed a sufficient uptake (>10 g/L serum IgG1), while 30.9 % of 

these calves had FPT, based on the Midland test. IgG concentrations 

were measured and the results were evaluated in other 97 serum 

samples of one-day-old newborn calves, using a single radial 

immunoassay (VMRD, Pullman, WA, USA). Based on the results of 

this method, 28 (28.9 %) of these calves had FPT, while 69 (71.1 %) 

had adequate immunoglobulin uptake. In 14 of the 28 FPT calves, 

diarrhoea or respiratory infection developed within 14 days. From 

the 69 calves that had normal IgG levels, only 12 (17.4 %) showed 

any symptoms of gastrointestinal or respiratory diseases until Day 14 

after birth. The difference between the two groups was statistically 

significant (P < 0.01). In conclusion, the different methods used for 

the IgG measurements gave similar results (30.9 % vs. 28.9 % FPT) 

in two consecutive years, which draws the attention to the 

importance of adequate immunoglobulin uptake and continuous 

control. 

This study was supported by OMFB-173-177/2006. 

Key words: calf, immunoglobulin, immunoassay 


187 Cooling Strategies to Improve Milk Production under Heat 

Stress Conditions 

L. Avendano 1 , D. Alvarez 1 , A. Correa 1 , R. Perez 1 , A. Algandar 1 , 

R. Diaz 1 , P. Robinson 2 , J. Fadel 2 

1 

Universidad Autonoma de Baja California, Animal Physiology, 

Mexicali, Mexico, Mexico 

2 

University of California, Animal Science Department, Davis, United 

States 

The aim of this study was to determine the best cooling strategy 

using spray and fans to improve milk yield and some metabolites in 

lactating Holstein cows under heat stress conditions. The experiment 

was carried out in a commercial dairy herd located in the Mexicali 

valley, Baja California, Mexico. Thirty two multiparous Holstein 

cows were blocked by milk yield and were assigned to one of four 

treatments: 1) control group, with cooling period before milking 

times; 2) in addition to 1, one cooling period at 11:00 h; 3) in 

addition to 1, one cooling period at 23:00 h; and 4) in addition to 1, 

two cooling periods, one at 11:00 h and the other at 23:00 

h. Duration of cooling time before milking was 30 min and for the 

additional cooling period 45 min. The cooling system was 

installed in the roof of the holding pen, before the milking parlor and 

consisted of four fixed fans of 90 cm with a mist head located in the 

center that delivered 28 L/h of water. Cows were milked twice a day 

(05:00 and 17:00 h) and they rotated in all treatments. Period in each 

treatment was 21 d and data were collected on the last week of each 

period. The study included from June 20 th to September 11 th of 2006 

and climatic variables were collected to estimate THI. Response 

variables were milk yield, fat and protein content, milk energy 

output, glucose, cholesterol, and triglycerides. The statistical model 

was a 4 X 4 Latin square that included block, period, treatment, cow 

nested into blocks, and block*treatment interaction. Orthogonal 

contrasts were used to compare treatment means. Average THI 

during the study was 83, with maximum of 88 and minimum of 78. 

The maximum temperature registered was 49 °C. These values are 

considered between moderate and severe heat stress conditions. Milk 

yield (21.12 vs. 19.10 ± 0.23 kg) and milk energy output (13.6 vs. 

12.6 ± 0.28 MCal) were higher (P0.05) in fat or protein content among treatments. However, 

glucose levels were higher (P0.05) among treatments. The 

cooling strategy with the best results was the one with more cooling 

periods (4), and represents an alternative to improved productivity of 

lactating Holstein cows in dry and hot regions. 

Key words: dairy cattle, hot weather, milk yield, Mexico 

188 Stress Responses in Adult Cattle due to Surgical Dehorning 

Using Three Different Types of Anaesthesia 

R. Lepkova, J. Sterc, V. Vecerek 

University of Veterinary and Pharmaceutical Sciences Brno, Brno, 


Objectives: Dehorning adult cattle is a surgical procedure causing 

distress of varying intensities that can be reflected in behavioural 

changes and alterations in plasma cortisol levels. Animal welfare 

considerations necessitate that the dehorning of cattle be done under 

conditions causing as little stress as possible. This study investigated 

the stress response to surgical dehorning in adult cattle, as expressed 

by blood cortisol levels and behaviour changes, using three types of 

anaesthesia. 

Materials and Methods: Stress responses during the dehorning 

process were evaluated in 18 Red Pied cows. The cows were divided 

into 3 groups of six and kept in tie-stall housing. Those in the first 

group were dehorned under general anaesthesia (GA) induced by 

intravenous administration of xylazine and ketamine. The second 

group was dehorned under sedation and local anaesthesia (SLA) 

induced by intramuscular administration of xylazine and local 


Animal Behaviour and Welfare 

anaesthesia with lidocaine. The third group was dehorned under local 

anaesthesia (LA) with lidocaine. Dehorning was performed with a 

foetotomy wire. Blood samples were taken 0.5 h before dehorning to 

determine cortisol levels, and, by means of a central venous catheter 

inserted into the jugular vein, during surgery, at 0; 0.5; 1; 1.5; 2; 2.5; 3; 

3.5; 4; 5; 6; 7 and 8 h post-surgery. Concurrently, occurrence of stress 

behaviours was assessed. Cortisol levels were measured by enzyme 

immunoassay (EIA). 

Results: The lowest mean peak levels of plasma cortisol (82.53 ± 6.04 

nmol l -1 ), the most rapid return of plasma cortisol levels to baseline 

values (1.92 ± 1.11 h), and the lowest occurrence of stress behaviours 

(2.38 - 5.83 %) were noted in the SLA group. The highest mean peak 

levels plasma cortisol (113.86 ± 25.65 nmol l -1 ), the slowest return of 

plasma cortisol levels to baseline values (3.83 ± 2.18 h) and the most 

frequent occurrence of stress behaviours (65.48 ± 28.72%) were 

observed in the LA group. There were significant differences between 

the SLA and LA groups in peak plasma cortisol levels (p = 0.011) and 

in occurrence of stress behaviours (p = 0.003). 

Conclusions: Sedation induced by intramuscular administration of 

xylazine in conjunction with local anaesthesia with lidocaine is 

considered the most suitable method of anaesthesia when dehorning 

adult cattle. Local anaesthesia with lidocaine alone was least suitable. 

189 Current Attitudes of European Veterinary Practitioners 

toward Pain and the Use of Analgesics in Cattle 

J. Huxley 1 , P. Van Dijk 1 , M. Gidekull 1 , R. Guatteo 1 , X. Manteca 1 , 

K. Müller 1 , B. Ranheim 1 , K. Touati 1 , S. De Vliegher 1 , H. Whay 2 

1 

University of Nottingham, School of Veterinary Medicine and 

Science, Nottingham, United Kingdom 

2 

University of Bristol, Clinical Veterinary Science, Bristol, United 

Kingdom 

As a result of their history as a prey species there has been a strong 

evolutionary pressure on cattle to mask signs of pain and its implied 

weakness. Consequently, recognising the signs of pain in this stoical 

species represents a significant challenge for veterinarians and may 

partially explain why the use of analgesics in cattle has lagged behind 

that of small animals and horses.A questionnaire designed to assess the 

attitudes of respondents to pain and the use of analgesics in cattle was 

distributed to 12,764 practitioners in nine European countries 

(Belgium, Denmark, France, Germany, The Netherlands, Norway, 

Spain, Sweden and the UK) between 2004 and 2006. The questionnaire 

collected data on demographics, availability of and attitudes towards 

analgesics, treatment regimes, estimated pain severity for a range of 

conditions and procedures in adult cattle and calves (assuming no 

analgesic agents were used) and their levels of knowledge in the field. 

Questionnaires from 2716 practitioners were returned (21.3%). 

Respondents graduated from one of sixty one Schools between 1957 

and 2006; 82% were male and 18% were female. Forty eight percent of 

respondents considered they had adequate knowledge in the area. The 

proportion of practitioners who stated they never used analgesic agents 

for a procedure or condition ranged from 1% for caesarean section to 

41.3% for dystocia in adult cattle and from 2.9% for umbilical hernia 

surgery to 55.1% following dystocia in calves. Based on the median 

results, mastitis (with clots only) and neck calluses were considered the 

least painful and claw amputation the most painful procedure or 

condition of adult cattle. Similarly the pain associated with dystocia 

was considered the least painful and surgical castration, burdizzo 

castration, distal limb fracture and umbilical hernia surgery the most 

painful procedure or condition of calves. The results generated from 

this study suggest that there are currently two principal factors 

hindering the use of analgesics in cattle: the limited number of 

analgesics (particularly local anaesthetics) licensed for use in food 

animal species in Europe and the belief amongst many practitioners 

that they have an adequate knowledge of pain management despite the 

fact that their use of analgesics in cattle is often limited. The authors 

would like to respectfully acknowledge the input of our late colleague 

Professor Ove Svendson of the Royal Veterinary and Agricultural 

University in Denmark. 

Key words: pain, analgesic, cattle, survey, welfare

190 Improving Welfare and Health of Dairy Cattle through 

Structured design of Husbandry Systems 

I. Van Dixhoorn 1 , P. Groot Koerkamp 2 

1 

Wageningen University, Animal Sciences Group, Lelystad, 


2 

Wageningen University, Farm Technology Group, Wageningen, 


Objective: Current husbandry systems for dairy cows are far from 

optimal. Reduced welfare, health problems and public discussions are 

typical problems encountered in traditional as well as in organic dairy 

production. An ideal husbandry systems should enable cows to express 

their natural behavior (limiting stress) and to maintain a good health 

status and enable the farmer to manage the herd and generate an 

income. The objective of this study was to design a husbandry system 

for a stable family herd of (dry) cows, calves, heifers and possibly a 

bull in which calves are not dehorned, calves can suckle, no external 

group changes occur and the needs of all animals are fulfilled. 

Materials and methods: The structured design method (Siers, 2004) 

was applied and the following steps were taken: Preliminary research: 

1 Analysis of the needs of the actors in the system; 2 System analysis; 

3 Identification of undesirables and set design objective; Problem 

definition: 4 Problem analysis; 5 List qualitative and quantitative 

aspects of the needs (Brief of Requirements); 6 Identification of key 

functions; Formulating solutions and concepts: 7 Find solutions; 8 

Combine solutions into structures and design concepts; 9 Evaluate 

concepts; Detailed design and shaping 

Results: In this paper we will focus on the results of specific steps. 

The brief of requirements focuses on the needs of all individual 

animals in the herd and the farmers‘ need to make the system 

controllable within the rules that are laid down by law. The animal 

needs are based on the 5 freedoms (Brambell) and the EU project 

Welfare Quality ® . A list of 13 key functions in the new husbandry 

system was identified. A selection: Individual cow recognition; • 

Provide and control feed per animal; • Access management; • Milk 

cows; • Enable suckling of calves. 

Four concepts were designed: • Low cost; • Easy manageable; • Most 

animal friendly; • Mix of 1 to 3. 

Elements of the final concept were: A central calf area to which only 

calves have access and are fed. A common space to which all animals 

have access at any time (calves can suckle their mothers here). All other 

animals are fed individually outside the common area, depending on 

lactation stadium. Individual “cow parameters” are measured. 

Conclusions: By using the structured design methodology an 

innovative design for complex dairy farming system can be made. 

Further improvement of the design and testing in practice will be done 

in close interaction with farmers, veterinarians, advisors and other 

stakeholders. 

Key words: welfare, innovative husbandry system, design 

191 Lesions at Medial Side of the Hock in Dairy Cattle 

M. Holzhauer 1 , G. Jousma 2 , JH. Hulsen 3 , PJ. Egberink 4 

1 

Animal Health Service Deventer, Ruminants, Deventer, Netherlands 

2 

Agriculture University, agriculture college, Wageningen, 


3 

Veterinary Practitioner, Veterinary Professional Services, Utrecht, 


4 

Veterinary Practice, Veterinarian, Utrecht, Netherlands 

Hock lesions in dairy cattle are a well known problem in all kind of 

clinical presentations in dairy industry. These lesions are especially 

known originating from the lateral side of the tarsus and varying 

from loss of hair until serious arthritis. These lesions seem to be 

related with the duration of indoor housing, type of bedding and stall 

design. Lesions at the medial side of the hock are a relatively new 

phenomenon in dairy cattle. We hypothesized that the lesions were 

caused by incongruities between udder shape and leg movements. 

Two pilot studies executed in a private practice in The Netherlands 

have investigated prevalence, risk factors and some aspects of 

aetiology of these lesions. In a first pilot study performed in 40 dairy 

herds (75-100% HF), a herd prevalence and a cow prevalence has 

been estimated of 100% and 14 % respectively and clear relation was 

found with parity and milk-production level. Compared to parity 2, 

for cows in parity 1 an OR has been estimated of 0.15 and for cows 

in parity 3 an OR has been estimated of 3.0; for cows with a 

production > 8000 kg, an OR has been estimated of 3.0. The lesions 

have been seen in cows during the second part of lactation and 

significant more frequently in cows with a low body condition 

score. In a 2 nd investigation a case-control study has been performed 

in herds with high and low prevalence. As a diagnostic tool chalk 

markings have been applied on the hind quarters of the udder and 

special attention has been paid to the origin of the litter also. A clear 

relation has been found between the presence of hock lesions at the 

medial side and skin lesions at the hind quarter of the udder and the 

deepness of the udders. Lesions were also significantly more 

frequently present in presence of strong grazing litter in the cow’s 

cubicles. Hock lesions both at the lateral and the medial side are 

related with diminished animal welfare and are potential entrances 

for all kind of bacterial infections with the risk of dissemination. 

Additionally, lesions at the medial side of the hock may also lead to 

secondarily infected udder skin and possible contamination of milk. 

For prevention of hock lesions at the medial side attention should be 

paid to the correct shape of the hind quarter of the udder, the use of 

soft dry bedding materials, prevention of loss of body condition and 

lameness. 

Key words: hock lesions, bedding, animal welfare 

192 Argentine Biotype Holstein and its High Plasticity, Based on 

the Production System 

D. Vacarezza 1 , A. Lopez 1 , M. Galicio 1 , D. Vacarezza 2 , 

E. Martinez 3 , C. Zenobi 4 , J. Lagger 5 , D. Chaves 5 , M. Glauber 5 , 

E. Villar 5 

1 

Facultad de Ciencias Veterinarias, Animal production, Buenos 

Aires, Argentina 

2 

Agriculture Faculty, Animal Welfare, Buenos Aires, Argentina 

3 

Facultad de Ciencias Veterinarias, Animal and Veterinary Science, 

Buenos Aires, Argentina 

4 

Animal Production Insititute, Anatomy, Histology and Ebryology, 


5 

Animal Production Insititute, Animal and Veterinary Science, 


The milky production in the Argentine Republic is obtained from 

Holstein argentine hybrids in a 95%. Having adapted to a pastoral 

system with supplementation, its productions are located in 

intermediate levels between New Zeeland type and those from USA; 

but with more plasticity than both of them. Our research was based 

on an analysis of the different production conditions of the last years 

in our country and compare them with the productions of the 

countries named before. The Argentine Republic suffered an 

economic crisis with a milk price disvalue that reached a 30% of its 

original value, earning the producers 9 dollar cents in 2002, with the 

beginning of a price recovery in 2003, reaching prices of 15-18 

dollar cents in 2004. We had to adapt us to both of this relative price 

conditions in such a brief time. We came to the conclusion that in 

favourable conditions our animals have a better respond than those 

from New Zeeland but lower than those from USA. Either wise, 

when the conditions turn unfavourable the respond of our cattle is 

superior than USA´s but inferior to New Zeeland´s. Several diets 

responds have been analysed, the effect over reproduction and the 

variation of the national production. It´s our belief that each country 

must take advantaged of the cattle that is better adapted to its 

ecologic conditions avoiding coping models. 

Key words: millk production Holstein Argentine system 

225 Survey Results on the Welfare of Dairy Cows in 250 Italian 

Farms 

A. Peli 1 , L. Scagliarini 1 , A. Serraino 2 , F. Giacometti 2 , S. Cinotti 1 , 

P. Famigli-Bergamini 1 

1 

University of Bologna, Veterinary Clinical Department, Bologna, 

Italy 

2 

University of Bologna, Public Health and Animal Science, Bologna, 

Italy 

Here we present the results of a survey performed to evaluate the 

welfare of dairy cows in more than 250 Italian farms (in the northern 

and southern Italy): our aim was to show the strong and the critical 

points regarding animal welfare and to verify the suitability of our 

method to extend the study on a wider number of (1027 other) farms. 

The survey has been conducted using a check list through 

Animal Behavior and Welfare 247

inspections, carried out by specifically trained technicians. The 

check list focused on more than 300 parameters, divided in 5 general 

sections, 6 specific sections for the different categories of animals 

and one section called “indicators”, that focused on the animals 

individual health status. These parameters have been evaluated on 

the basis of specific limits defined in a manual for the technicians 

and were classified as “conforming” or “non-standard (not 

conforming)” referring to the law in force, or as “substandard”, 

“adequate”, “good”, “excellent”, on the strength of the scientific 

data available in the literature. The parameters not objectively 

measurable (about 100) have been classified as “descriptive” and 

thus not used in the analysis. The farms were classified as “small” 

(less than 40 cows), “medium” (from 40 to 79 cows) and “big” (more 

than 80 cows).The parameters included in the analysis were about 

200 but only few of them (

227 Genetic Relationship of Body Energy and Blood Metabolic 

Traits with Reproduction and Reproductive Health in Dairy 

Cows 

G. Oikonomou 1 , G. Arsenos 1 , G. Valergakis 1 , A. Tsiaras 2 , 

D. Zygoyiannisv, G. Banos 1 

1 



2 

Vivartia Avee, Athens, Greece 

The objective of this study was to investigate the genetic relationship of 

various energy balance indicators with first and second lactation 

reproduction and reproductive health of dairy cows. 497 primiparous 

Holstein cows raised on a large commercial farm in Greece were 

included in the study. The energy balance indicators used were body 

condition score (BCS), total body energy content (EC), cumulative 

effective energy balance (CEEB), and blood serum levels of glucose, ‚hydroxybutyrate 

(BHB) and non-esterified fatty acids (NEFA), measured 

throughout first lactation. Insemination, conception and interval traits 

were recorded for the evaluation of cow reproductive performance. 

Incidence of clinical metritis, diagnosed by the farm’s veterinarians, was 

also recorded. Random regression models were used to calculate weekly 

animal breeding values for first lactation BCS, EC, CEEB, glucose, BHB 

and NEFA. Reproductive records of individual cows were then regressed 

on these breeding values to derive estimates of genetic correlations. 

Several significant (P

The origin of Hungarian Grey cattle breed is still controversial. The 

theory of Asiatic origin and the theory of European domestication 

are the two most accepted ones. The domestication is more 

supported by archeozoological findings, but the Asiatic origin is 

supported also by some mtDNA sequence data of previous study of 

Beja-Pereira (2006). Our sampling method has considered the 

history of breed, because it has a serious bottle neck after the World 

War II, and in that time the nucleus herd had only 150 cows. 

According to this genetic drift, it is very important to investigate the 

mitochondrial lines of these founders directly in the nucleus herd, 

and not to take samples from commercial herds without pedigree 

information. 80 mitochondrial D-loop samples of founder lines were 

sequenced between the 15904 and 16308 positions of mitochondrial 

DNA. We have found 37 haplotype, and the most part of the 

sequences (90%) belong to the most common European T3 

haplogroup (Troy 2001). Our results support the domestication 

origin from European wild oxes of Hungarian Grey breed. 

Comparisons of the results with previous studies emphasize the 

importance of sampling practice for phylogenetic studies. Due to the 

maternal inheritance of mtDNA and differences between natural 

populations and breeds a stratified sampling method is needed. From 

the mitochondrial sampling point of view the representative sample 

of a breed must contain as many samples of different mtDNA lines 

of founder cows as possible, and without the complex investigation 

of pedigree the value of sample set is often questionable. The other 

weak point of phylogenetic sampling practice is the small number of 

samples. When the pedigree structure is known the statistically 

acceptable minimum size of sample is depending on the number of 

founders. Extreme low number of studies contains information about 

these sampling techniques. 

Key words: Hungarian Grey, mtDNA, founder, domestication, 

phylogenetics 

231 Growth Performance, Carcass Traits and Meat Quality of 

Young Bulls of Four Dual Purpose Breeds of the Italian Alps 

M. Brscic, M. Colautti, F. Gottardo, B. Contiero, G. Cozzi 

University of Padua, Animal Science, Legnaro, Italy 

In the last decades, specialized dairy cattle breeds have replaced the 

ancient strains of the Italian Alps leading to a decline of the 

landscape and reducing biodiversity. The study aimed to assess the 

aptitude towards meat production of bulls belonging to 3 local 

breeds raised in the Italian Alps: Burlina (BUR), Alpine Grey (AG) 

and Rendena (REN) comparing them to a cosmopolite dual purpose 

breed, the Italian Simmental (IS). A fattening trial used 48 calves, 12 

per breed (initial body weight: 204±6 kg) allotted according to breed 

in 12 straw bedded pens of 4 animals. Bulls were fed ad libitum a 

total mixed ration (DM:63.3%; CP:13.4%DM; NDF:33.5%DM; 

Starch:30.8%DM). Body weights were recorded at start and end of 

the trial in order to calculate average daily gains (ADG). Pen DM 

intake was recorded weekly and feed efficiency (intake/daily gain) 

was calculated. Health status of bulls was monitored throughout the 

fattening period. Slaughtering occurred when bulls reached the 

optimal finishing status. At the slaughterhouse, carcass weight and 

dressing percentage were measured and their conformation 

(SEUROP) and fatness were graded. Weight of head, skin and 

forestomach content were also recorded. Samples of Longissimus 

thoracis, excised from the 7th to the 9th rib after an ageing period of 

11 d under vacuum, were analyzed for chemical composition, 

instrumental colour, cooking losses and shear force. Data were 

submitted to statistical analyses adopting a linear model considering 

the effects of breed and pen(breed). Bulls of IS and BUR showed a 

longer fattening period (301 and 297 d) in comparison to AG and 

REN (275 and 276 d). The IS showed the highest slaughter weight 

(624 kg) and ADG (1399 g/head/d) in comparison to the other breeds 

which had similar values. Feed efficiency resulted similar across 

breeds due to the higher DM intake of IS bulls. Carcass weight was 

significantly higher for IS. On the contrary, there were no 

differences among breeds for dressing percentage likely due to a 

heavier trimmed bone or fat for IS since head, forestomach content 

and skin had the same or lower weights than other breeds. Shear 

force was the only meat quality trait affected by breed, lower for 

BUR (3.17 kg/cm 2 ). In conclusion, compared to cattle breed with a 

good vocation for meat production like IS, the 3 Alpine breeds gave 

satisfying growth and slaughter performance sustaining their dual 


purpose. This should encourage mountain farmers to raise the males 

for meat production as a tool to increase profits. 

Key words: beef cattle, dual purpose breeds, growth performance, 

slaughter traits, meat quality 


1204 Single Nucleotide Polymorphism (SNP) Analysis of Bacterial 

PAMPs Associated with Toll Like Receptors (TLR1, TLR2 

and TLR4) in Autochthonous Central European Cattle 

M. Bhide 1 , R. Mucha 1 , I. Mikula 1 , L. Kisova 1 , M. Novak 2 , I. Mikula 1 

1 University of Veterinary Medicine, Laboratory of Biomedical 

Microbiology and Immunology, Kosice, Slovakia 

3 Institute of Neuroimmunology, Laboratory of Biomedical 

Microbiology and Immunology, Bratislava, Slovakia 

Toll like receptors (TLR1, TLR2 and TLR4) are known to recognize 

the bacterial PAMPs (Pathogen associated membrane proteins), and 

initiate the innate immune signalling. Recently, close linkage 

between mutations/SNPs in the protein coding region of TLRs and 

altered responsiveness to bacterial PAMPs has been described. Here 

we attempted to map SNPs in the coding regions of TLR1, TLR2 and 

TLR4 in autochthonous breeds from central European cattle. 

Population of 559 cattle consisting of Slovak spotted cattle (n=10), 

Polish red (n=64), Pinzgauer (n=247), Slovakian Simmental (n=206) 

and Dark brown Carpathian (n=32) breeds. TLR gene segments were 

amplified by PCR using genomic DNA as template. PCR product 

was then subjected for single strand conformation polymorphism 

analysis to group the samples according to similar SSCP profiles 

(Genotypes). Representative sample from each genotype was 

sequenced and SNPs were validated by SeqScape 2.1 software. Only 

two TLR4 genotypes occurred in the population, and interestingly 

both was in homozygous state. This indicates highly conserved 

nature of TLR4 gene. Although, only three TLR2 genotypes were 

found, two of them were heterozygous. Novel missense SNPs found 

in bovine TLR2 gene were Ile680(Ile, Leu), Glu738(Asn, Ser) and 

Lys754(Lys, Arg) with 0.09, 0.04 and 0.04 frequencies respectively. 

TLR1 was the most heterogeneous in the population with 14 SNPs, 

three silent and six missense distributed in six genotypes. The TLR1 

missense SNPs were Ser216(Ser, Phe), Leu226(Leu, Phe), 

Ser228(Ser, Phe), Ser234(ser, Phe), Thr236(Thr, Ile) and 

His244(Gln, His). Till to date, SNPs like Pro681His and Arg677Trp 

in TLR2 gene, and Asp299Gly in TLR4 gene are reported as SNPs 

that cause increased susceptibility (risk SNPs) of host to 

mycobacterial and streptococcal infections. Many novel mutations 

and SNPs are being investigated in vitro and in vivo to assess their 

effect on disease resistance (linkage studies). SNP mapping 

presented by us in TLR genes, the key genes of innate immunity, 

make benchmarking for the linkage studies. Culling of the animals 

having risk SNPs can be the effective tool in animal breeding to 

increase the natural resistance of heard against selected bacterial 

infections. 

Key words: SNPs, TLR, natural resistance, breeding, PAMPs 

1205 Selection of Mathematical Model for Describing Lactation 

Curve Using Milk Records of a Single Herd of Holstein Cow 

in Iran 

A. Boustan, M. Moradi Shahrebabak, A. Nejati javaremi, 

M. D. Banadaki 

University of Tehran, Animal Science, Karaj, Iran 

Test day milk records of a Holstein dairy herd which were collected 

during years 2004 to 2006 of 142 cow in 1st lactation and 140 cow in 

2st lactation were used in this research. There were real milk yield of 

each cow in each day of lactation in this farm and consequently we 

have real total milk yield of each cow. These yields used for 

comparing functions. Six different mathematical functions include 

Incomplete gama function. Third order legendre polynomial 

function, Polynomial regression function, Inverse polynomial 

function, Wilmink function and Mixed log function were fitted to 

data. Each function was fitted to data of each cow by a program in 

visual basic (monthly record of each cow was used in this stage). 

This program calculated adjusted R-square and 305-day milk yield

(5-310 th day milk yield) for each cow. Functions were compared 

based on adjusted R-square and squared error of estimation. Squared 

error of estimation calculate by squaring difference between milk 

yield that was estimated for each cow by each function and real milk 

yield for this cow. We have an squared error for each cow for each 

function. Functions were ranked by quad test on squared error. This 

test made clear rank and significance of difference between rank of 

functions. Mean of adjusted R-square for each function for each 

lactation were calculated. Based on results of this research 

Incomplete gama, Mixed log, Inverse polynomial, Wilmink, Third 

order legendre polynomial and Polynomial regression function had 

1 st to 6 th rank in 1 st lactation respectively. Incomplete gama, Mixed 

log, Wilmink, Third order legendre polynomial, Inverse polynomial, 

Polynomial regression function had 1 st 

to 6 th 

rank in 2 nd 

lactation respectively. Incomplete gama function had the first rank 

for estimation of milk yield in each lactation but its difference by 

Inverse polynomial function, Wilmink function and Mixed log 

function was not significant in 1 st lactation and its difference by 

Inverse polynomial function, Wilmink function, Mixed log function 

and Third order legendre polynomial function was not significant in 

2 nd lactation. Incomplete gama function also had the highest adjusted 

R-square in both lactation. Based on the results of this research 

incomplete gamma function works better than other functions used 

in this research. 

Key words: Holstein cows, lactation, fitting, squared error, function 

1206 Transcriptomic Pattern of Skeletal Muscle in Meat Breed 

Bulls 

T. Sadkowski 1 , M. Jank 1 , J. Oprzadek 2 , L. Zwierzchowski 2 , 

T. Motyl 1 

1 Warsaw University of Life Sciences - SGGW, Physiological 

Sciences, Warsaw, Poland 

2 Polish Academy of Science, Institute of Genetics and Animal 

Breeding, Jastrzebiec, Poland 

For many years there have been a lot of studies conducted aiming to 

identify the genes and proteins responsible for phenotype of animals 

belonging to specific directions of animal production. The aim of the 

present study was to compare the transcriptomic profile of skeletal 

muscle (m. semitendinosus) of bulls of pure meat performance 

(Limousine, Hereford) as well as bulls of mixed meat-milk 

performance (polish red) with bulls of pure milk performance 

(Holstein-Friesian type black-and-white). The studies were 

performed on bovine cDNA microarrays (the NBFGC EST 

collection; Michigan State University, USA) containing 18263 

cDNA probes representing different bovine tissues in different 

physiological states. In the present study we identified 117 genes for 

limousine breed, 210 genes for Hereford breed and 221 genes for 

polish red breed which expression differed (increased or decreased) 

at least 2-times when compared to Holstein-Friesian breed. We 

identified also 48 genes which expression was correlated for breeds 

of pure meat performance (Limousine and Hereford). These genes 

were accepted as genes responsible for transcriptomic pattern of 

skeletal muscle with fast gain of muscle mass. 

The results of microarray experiments were verified by evaluation of 

expression of four randomly selected genes belonging to 

transcriptomic pattern (TRIP12, c-erbB3, MRPS30, PYCRL) with 

real-time PCR and the result obtained were in accordance with 

results of microarray. Using Panther software we performed 

functional analysis of genes belonging to transcriptomic pattern in 

order to establish their role in particular biological processes and 

molecular function. Total 14 out of 48 genes were involved in 

protein metabolism and modification. With respect to molecular 

function we identified three predominating groups of genes: one 

group of 10 genes which protein products bind with nucleic acids, 

second group of 6 genes which products are regulatory proteins and 

third group of 4 genes for kinases. Although the role of majority of 

genes belonging to transcriptomic pattern as well as their 

interactions are still not clear, the significant difference in their 

expression between skeletal muscle of cattle of pure meat 

performance (Limousine, Hereford) and pure milk performance 

(Holstein-Friesian type black-and-white) indicates their involvement 

in growth, development and metabolic processes in skeletal muscle 

of meat breed bulls. 

Key words: skeletal muscle, transcriptomic pattern, bovine, 

microarray 

1207 Study of Genetic Diversity in Iranian Mouflon Population 

Based on Microsatellite Marker 

S. Ghovvati 1,2 , M. Nassiri 1 , F. Eftekhari-Shahroudi 1 , M. Soltani 1 , 

A. Javadmanesh 1 

1 

Ferdowsi University of Mashhad, Animal and Poultry Science, 

Mashhad, Iran 

2 

Biotechnology Research Institute, North Region, Animal 

Biotechnology Group, Rasht, Iran 

Studying of biodiversity is a basic approach in order to make suitable 

decisions to protect endangered species. For this purpose, 75 biological 

samples were prepared from Tandureh National Park’s mouflon 

population (Khorasan, Iran). DNA was extracted from blood, meat, 

hair and bone samples. Total DNA was extracted from each sample by 

using Wizard SV Genomic DNA Purification System kit (Promega, 

California, USA) based on silica gel columns. The kit was used 

following the manufacturer’s instructions with minor changes to adapt 

the procedure to a high quantity of sample. The quantity and quality of 

extracted DNA were estimated with the spectrophotometer (Biometra, 

Germany). Ten microsatellite loci with their primers were selected, 

based on Food and Agriculture Organization (FAO 2004, United 

Nations) recommendations. PCR amplified products were separated by 

electrophoresis on non-denaturing 8% polyacrylamide gel stained with 

AgNO 3. All studied markers were polymorphic (5 to 8 allele). Totally 

61 allele with a mean number of 6.1 allele per locus were detected. The 

highest number of observed alleles belonged to MAF36 (8 allele) and 

the lowest was for OarFCB304 and OarHH47 (5 allele). Mean 

observed heterozygosity and mean expected heterozygosity were 

0.5842 and 0.7849, respectively. Mean of F is index in this study was 

about 0.2508. The minimum and maximum diversity for (Shannon 

index) were 1.5112 and 1.7231 for OarHH47 and MAF36, 

respectively. Results revealed that inbreeding level in Tandureh’s 

mouflon population is increased dangerously. These findings indicate 

the reduction of genetic diversity in this population that is so grave for 

survival of this species. 

Key words: genetic diversity, Iranian mouflon, Tandureh National 

Park, PCR, microsatellite 

1208 Researches Concerning the Comparison of Milk Yield 

Between Pinzgau Breed Imported from Austria and Pinzgau 

of Transylvania 

I. Gilca 1 , C. Pascal 2 , M. Ivancia 3 , B. Pasarin 4 

1 University of Agricultural Sciences and Veterinary Medicine, 

Animal Husbandry and Dairing, Iasi, Romania 

2 University of Agricultural Sciences and Veterinary Medicine, Sheep 

and Goats Technology, Iasi, Romania 


Animal Breeding, Iasi, Romania 


Animal Husbandry Department, Iasi, Romania 

The main aim of this study was to compare the milk performance 

realized by cattle of Pinzgau breed (PZ) imported from Austria and 

Pinzgau of Transylvania (PZT) - indigenous breed. The analysis was 

performed in two herds at the first and second lactations. In herd 1, 

the imported dairy cattle (P) were compared with contemporaries 

indigenous breed (PZT) calved in the same period. Both breeds were 

kept in the same conditions and with the same feeding. The first 

lactation records were analyzed according to the following linear 

model: Y ij =+ H i + C j + e ij. The second lactation records were 

analyzed according to the following linear model: Y ijk =+ H i + C j + 

J k + e ijk. The difference in milk production between Pinzgau and 

Pinzgau of Transilvania breeds, at first and second lactations, were 

not statistically significant. The Pinzgau breed cattle achieved higher 

fat, protein and lactose percentage at the first lactation (4.22% and 

3.96%, 3.39% and 3.21%, respectively 4.63% and 4.51%) and at the 

second lactation (4.28% and 4.01%, 3.41% and 3.23%, respectively 

4.58% and 4.46%). These differences were statistically highly 

significant (P

University of Warmia and Mazury in Olsztyn, Department of Animal 

Genetics, Olsztyn, Poland 

To determine the actual carrier frequency of the CVM (Compex 

Vertebral Malformations) in a population of Polish Holstein-Friesian 

Black-and-White cattle, a study was undertaken that examined 355 

proven bulls (active within 2001 - 2007) used by 4 domestic artificial 

insemination companies. A total of 69 T/G heterozygotes were 

diagnosed (19,4%), being mostly sons of known carriers of the CVM 

defect. Identification of a polymorphism in a bovine solute carrier 

family 35 member 3 gene, named SLC35A3, was carried out with the 

use of a new PCR-SSCP method (polymerase chain reaction - single 

stranded conformation polymorphism) which, due to ease of use and 

high reliability, can be applied in widespread screening programs. 

All CVM carriers were diagnosed once more by licensed allelespecific 

- PCR method. Complete accordance between the numbers 

275 Phenotypic and Genotypic Identification of Methicillin 

Resistance of Staphylococci Isolated from Mastitic Milk 

Samples 

B. Sareyyupoglu 1 , Z. Cantekin 2 , M. Akan 1 

1 Faculty of Veterinary Medicine, Ankara University, Department of 

Microbiology, Ankara, Turkey 

1 Faculty of Veterinary Medicine, Mustafa Kemal University, 

Department of Microbiology, Hatay, Turkey 

In this study, phenotypic and genotypic identification of methicillin 

resistance of staphylococci isolated from mastitic milk samples 

collected from 3 different districts (Polatli, Çubuk and Haymana) of 

Ankara were investigated. A total of 83 isolates, distribution of 

which was, 22, 29, and 32 from Polatli, Çubuk and Haymana, 

respectively, were used in the study. Isolates were identified with 

conventional identification tests. Kirby-Bauer disc diffusion method 

using oxacillin (1 µg) discs were used for the phenotypic 

identification of methicillin resistance. Furthermore, susceptibilities 

of isolates to amoxycillin+clavulanic acid (30 µg), amoxycillin (10 

µg), ampicillin (10 µg), and penicillin (10 µg) were also investigated 

in the study. For the genotypic determination of methicillin 

resistance, a multiplex-PCR technique co-amplifying the specific 

fragment of 16S rDNA in staphylococci, mecA gene encoding the 

methicillin resistance, and femA gene discriminating S. aureus from 

other staphylococci were used. Moreover, for the determination of 

beta lactamase activity, nitrocefin sticks and a PCR technique 

detecting blaZ gene were used. Following the tests, distribution of 

oxacillin resistance were detected 31.8%, 58.6% and 0%, 

respectively in isolates from Polatli, Çubuk and Haymana, while, 

mecA gene was detected 31.8%, 31% and 3.1% in these districts. 

BlaZ gene was detected 59.1% in Polatli isolates, 41.4% in Çubuk 

isolates and 59.4% in Haymana isolates. MecA gene was detected in 

17 (20.5%) out of 83 isolates, while 10 (58.8%) of these were 

determined to be methicillin-resistant S. aureus (MRSA). As a 

conclusion, multiplex-PCR technique was found to be a fast, specific 

and reliable technique for the simultaneous detection of methicillin 

resistant isolates and the discrimination of S. aureus isolates from 

other staphylococci. 

Key words: mastitis, methicillin resistance, multiplex-PCR, 

staphylococci 

276 Extended Biofilm Susceptibility Assay for Staphylococcus 

aureus Bovine Mastitis Isolates: Evidence for Association 

between Agr-type and Biofilm Susceptibility. 

M. Melchior 1,4 , M. Van Osch 1 , T. Lam 2 , W. Gaastra 3 , 

J. Fink-Gremmels 1 

1 

Utrecht University, Department of Pharmacology and Toxicology, 

Utrecht, Netherlands 

2 

GD Animal Health Service, Udder Health Service, Deventer, 



of CVM carriers genotyped by both methods was observed. Due to 

high breeding value of selected CVM carrier bulls’ - breeders 

hesitate whether to inseminate cows with them or not. In “milk 

productive farms” the risk to produce potential CVM homozygotes 

is very low if mating will take into account CVM status of their 

ancestors, already available in breeding documents. Special attention 

should be paid in “breeding herds” producing elite bulls and dams. 

Mating these animals should be assisted with continuous diagnostics 

to remove gaps in CVM status of pedigree and active education 

carried out by inseminators and A.I. dealers. Only CVM-free young 

bulls are allowed to enter progeny testing program in A.I. station. 

This strategy should reduce the incidence of CVM carriers in very 

short time. Closer international integration of national associations 

of Holstein cattle breeders and coordination of genetic defects 

discovery programs is necessary to decrease the number of carriers 

and affected animals at a faster rate. 

Good Veterinary Practice (Antibiotic Resistance) 

3 

Utrecht University, Infectious Diseases and Immunology, Utrecht, 


4 

Central Veterinary Institute, Bacteriology and TSE's, Lelystad, 


Staphylococcus aureus is one of the most prevalent causes of bovine 

mastitis. The antimicrobial treatment of this disease is currently 

based on antimicrobial susceptibility tests according to CLSI 

standards. However, various studies have shown that there is a 

discrepancy between the results of this standard susceptibility test 

and the actual cure rate of the applied antimicrobial treatment. 

Increasing evidence suggests that biofilm formation by S. aureus is 

associated with this problem. Previous data obtained with a limited 

number of strains revealed that the extended biofilm antimicrobial 

susceptibility assay allows differentiation between strains, which 

cannot be derived from a standard susceptibility test or from a 24hour 

biofilm susceptibility test (Melchior, et al., 2007). The 

objective of this study was to test a representative collection of S. 

aureus bovine mastitis strains in the extended biofilm susceptibility 

assay. Based on the results from a previous study with the same 

collection of strains the effects of Agr-type (Accessory Gene 

Regulator gene) and the presence of IS257 (Insertional Sequence 

257) in Agr-type II strains on the biofilm susceptibility were also 

studied. The Agr locus of S. aureus controls the expression of most 

of the virulence factors it represses the transcription of a number of 

cell wall-associated proteins, and activates several exoproteins 

during the post exponential phase (Bronner, et al., 2004). The IS257 

gene has been related to biofilm formation in vitro (Cramton, et al., 

1999) and was found earlier in 50% of the Agr-type II strains.The 

results of this study revealed differences in biofilm susceptibility 

between Agr-type I, III and IV strains and Agr-type II strains which 

were also dependent on the antimicrobial used. The presence of 

IS257 in Agr-type II strains has a marked effect on the in vitro 

biofilm density and on antimicrobial susceptibility of these strains 

growing in biofilm. The decreasing antimicrobial efficacy with older 

biofilms and the increasing efficacy with longer duration of 

antimicrobial challenge were shown. These data explain the better 

therapy results for penicillin susceptible strains in vivo obtained in 

several epidemiological studies (Sol, et al., 1997, Taponen, et al., 

2003). The data presented here also offer an explanation for the 

higher efficacy of early antimicrobial treatment and for treatments of 

longer duration in bovine mastitis S. aureus infections described in 

the literature (Sol, et al., 2000). 

Key words: mastitis, antimicrobial susceptibility, bacterial biofilm, 

Staphylococcus aureus, Agr- typing 

277 Longitudinal Study on Antimicrobial Resistance Patterns of 

Salmonella spp. and E. coli obtained from Dairy Cattle in 

Colorado, USA 

A. Villarroel 1 , D. Dargatz 2 , M. Salman 3 , S. Ladely 4 , 

P. Fedorka-Cray 4

1 

Oregon State University, College of Veterinary Medicine, Corvallis, 

United States 

2 

USDA-APHIS-VS-CEAH, Fort Collins, United States 

3 

Colorado State University, Animal Population and Health Institute, 

Fort Collins, United States 

4 

USDA-ARS, Athens, United States 

Objective of study: The objective of this study was to describe 

antimicrobial resistance patterns of Salmonella spp. and non-type 

specific Escherichia coli isolated from the same animals and same 

environmental areas over time. 

Materials and Methods: Sampling occurred at a local dairy over a 12 

month period, at 8 week intervals. Samples were collected from animal 

feces (cows and calves), feed, drinking water, hospital milk, colostrum 

and alleyway flush water. 

Results: Salmonella serotypes recovered from environmental and fecal 

samples collected from the same groups of animals differed. 

Salmonella serotypes were different between calves and cows. In 

addition, antimicrobial resistance patterns of both Salmonella and E. 

coli were different. 

Conclusions: Based on our data, we conclude that under field 

conditions Salmonella and E. coli may not exchange genetic material as 

readily as previously suggested. These results may be partially due to 

effective biosecurity procedures on this particular dairy, avoiding 

cross-contamination between cow and calf-areas. 

Key words: antimicrobial resistance, dairy cattle 

278 Subconjunctival (SC) Injection of Penicillin-Streptomycin in 

Keratoconjunctivitis of Calves and Adult Cattle - an Efficient, 

Economic Approach 

E. Bani Hassan 1 , T. Taghipour Bazargani 1 , R. Rahchamani 1 , 

H. Sharifi 2 , A. Hosseinzadeh 3 , I. Ashrafi Tamay 1 , Sh. Moaddab 1 , 

E. Torki 1 

1 



2 


of Food Hygiene, Tehran, Iran 

3 

Urmia University, Department of Veterinary Microbiology, Urmia, 

Iran 

Objectives: To determine the efficacy of IC-therapy and comparing 

this method to other common therapies. 

Materials and methods: The study was carried out as a pilot in two 

dairy farms and after approval of the efficacy of subconjunctival 

therapy we used this method to as the routine treatment for 

conjunctivitis and keratoconjunctivitis. At the first stage we showed 

that the single SC injection of penicillin-streptomycin, showed 

completely superior results compared to other methods including 

systemic antibiotic therapies with oxitetracycline, oxitetracycline plus 

tylosine, florefenicole, and even systemic injection of penicillinstreptomycin 

(p=0.0001).In the next step we choose the method as the 

usual treatment for the disease in both calves and adult cattle. This 

resulted in deletion of the problem of blindness and growth retardation 

due to keratoconjunctivitis (P


1225 The Effect of Herbal Medicine on Cow Dramatic and Vagina 

Infection Treatment 

A. Ahadi 1 , M. Sanjabi 2 , M. Moeini 3 

1 IROST, Animal Science, Karaj, Tehran, Iran 

2 IROST, Animal Science Department, Karaj, Tehran, Iran 

3 Razi University, Animal Science Department, Kermanshah, Iran 

The role of medicinal plants is considered recently in animal treatment. 

Iran has been endowed with a rich flora variety of plant and herbal due 

to its varied geographical and climatic condition. A large number of 

indigenous medicinal plants have been reported to possess woundhealing 

properties. Studies have shown that herbal plants such as 

Matricaria chamomilla; Achillla millefolium have anti inflammatory, 

anti bacterial and anti fungal activity in herbal medicines (Atta 1998, 

Dixit 1998). RHAM is an indigenous preparation, whose clinical 

effects on wound healing have been studied. In this study Ethanolic 

extract of plants of Maticaria chamomilla, Achilla, Rosa 

damascenamill and lawsonia (RHAM) were prepared and different 

combination gels were formulated. 54 infected dairy cows were 

selected from 4 herds in suburb of Tehran. The infected cows were 

treated by using (RHAM) during three months treatment. More than 

79% of the infected cows indicated a reliable recovery and the RHAM 

shows the potential therapeutic effect on infections. It can be concluded 

that the effect of RHAM on healing of skin, wound injuries and vagina 

infections was noticeable, quick and positive. 

Key words: herbal plants, medicines, dairy cows, vagina infections 

1227 Genotyping and Biofilm Formation in Different Growth 

Media of Staphylococcus aureus Bovine Mastitis Isolates: 

Evidence for Lack of Penicillin-resistance in Agr-type II 

Strains 

M. Melchior 1,3 , M. van Osch 1 , R. Graat 1 , E. Van Duijkeren 2 , 

M. Mevius 3 , M. Nielen 4 , W. Gaastra 4 , J. Fink-Gremmels 1 

1 Utrecht University, Veterinary Pharmacy and Toxicology, Utrecht, 

The The Netherlands 

2 

Utrecht University, Infectious Diseases and Immunology, Utrecht, 

The The Netherlands 

3 

Central Veterinary Institute, Bacteriology and TSE's, Lelystad, The 

The Netherlands 

4 Utrecht University, Farm Animal Health, Utrecht, The The 


The increasing evidence regarding the role of biofilm formation in 

bovine Staphylococcus aureus mastitis infections led to further 

investigations on biofilm formation in standard growth medium and 

bovine milk whey. For a collection of 99 recently isolated and 

historical S. aureus strains, the biofilm forming ability in both growth 

media was correlated with the presence of the ica-, bap-, and IS257 

genes. These genes have previously been found to be correlated with 

biofilm formation in human S. aureus isolates. All strains were 

genotyped with respect to Agr-type and Agr-subtype, and the presence 

of the resistance genes blaZ and smr by PCR. The prevalence of Agr 

types and investigated genes and the correlations hereof with the results 

of the biofilm assays were statistically evaluated. The data showed a 

very strong correlation between Agr-type I and penicillin-resistance in 

the bovine S. aureus mastitis strains. None of the Agr-type II strains 

were found to contain penicillin resistance genes. The Agr-type also 

had a significant effect on biofilm formation, however contrary to 

human isolates there was no significant effect for ica- and IS257 genes 

on biofilm formation. The bap gene was not found in any of the 

investigated strains. The presence of both biofilm related genes and 

resistance genes showed a high correlation with the Agr-type of the 

strains. These results indicate that the most prevalent Agr-types in S. 

aureus bovine mastitis, Agr-type I and II, might be regarded as 

different subspecies. The extremely high correlation between Agr- type 

II and penicillin susceptibility strongly suggests that these strains are 

not perceptive for the blaZ genes. 

Key words: bovine mastitis, penicillin susceptibility, bacterial biofilm 

1228 Comparison of Antimicrobial Resistance Patterns of 

Salmonella spp. and E. coli Obtained Using the Same Fecal 

Samples from Dairy Cattle in Colorado, USA 



P. Fedorka-Cray 4 

1 Oregon State University, College of Veterinary Medicine, Corvallis, 

United States 

2 USDA-APHIS-VS-CEAH, College of Veterinary Medicine, Fort 

Collins, United States 

3 Colorado State University, Animal Population and Health Institute, 


4 USDA-ARS, Athens, United States 

Objective of study: The objective of this study was to compare 

antimicrobial resistance patterns of Salmonella spp. to those of nontype 

specific Escherichia coli isolated from the same cattle. 

Materials and methods: Fecal samples were collected from calves 

and cows at a local dairy over a 12 month period, at 8 week intervals. 

Each fecal sample was cultured in specific media for the isolation of 

Salmonella spp. and non-type specific Escherichia coli. 

Results: Of a total of 1,665 fecal samples collected, only 43 had 

concurrent isolation of Salmonella spp. and E. coli. Of these, 14 

samples had both a resistant Salmonella spp. isolate and a resistant E. 

coli isolate. None of these isolates had the same resistance phenotype. 

Conclusions: Based on our data, we conclude that under field 

conditions Salmonella and E. coli may not exchange genetic material as 

readily as previously suggested based on in vitro studies. 

Key words: antimicrobial resistance, dairy cattle 

1229 Factors Associated with Isolation of Antimicrobial Resistant 

Salmonella spp. and E. coli from Dairy Cattle in Colorado, 

USA 


P. Fedorka-Cray 4 

1 


United States 

2 

USDA-APHIS-VS-CEAH, Fort Collins, United States 

3 

Colorado State University, Animal Population and Health Institute, 


4 

USDA-ARS, Athens, United States 

Objective of study: The objective of this study was to evaluate 

possible factors associated with isolation of antimicrobial resistant 

Salmonella spp. and non-type specific Escherichia coli from fecal 

samples from dairy cattle (calves and cows). 

Materials and methods: Fecal samples were collected from calves 

and cows at a local dairy over a 12 month period, at 8 week intervals. 

Each fecal sample was cultured in specific media for the isolation of 

Salmonella spp. and non-type specific Escherichia coli. Factors 

evaluated included age, housing, location, production level, 

antimicrobial treatment and sampling event (month of the year). 

Multivariate hierarchical multilevel analysis was performed to evaluate 

factors, forcing antimicrobial treatment into the model. 

Results: The factor most strongly associated with isolation of resistant 

Salmonella spp. in calves was housing in individual hutches (after 

adjusting for age). Treatment within 3 weeks of sampling and young 

age were the two factors most strongly associated with isolation of 

resistant E. coli in calves. Lactation number and medium to high milk 

production were the two factors most strongly associated with isolation 

of resistant Salmonella spp. in adult cows. Treatment with 

antimicrobials was the factor most strongly associated with isolation of 

resistant E. coli in adult dairy cows. 

Conclusions: Based on our data, we conclude that antimicrobial 

resistance in Salmonella and E. coli isolated from dairy cattle is a 

multifactorial condition influenced by previous antimicrobial treatment, 

management factors and inherent characteristics of the cattle (e.g. age). 

Key words: anitmicrobial resistance, dairy cattle 

1230 Evaluation of Antimicrobial Effects of 4 Medicinal Plants 

against Salmonella typhimurium and Comparision them 

with Common Antibiotics in Veterinary Medicine 

T. Zahraei Salehi 1 , M. Dakhili 2 

1 

Faculty of Veterinary Medicine, University of Tehran, Microbiology, 

Tehran, Iran 

2 

Faculty of Medical Science, Microbiology, Tehran, Iran 

Background: Thyme (Zataria multiflora Bois.), Penny voyel 

(Mentha pulegium L) Origanum (Origanum vulgare L), Fennel

(Foeniculum vulgare Miller) are an Iranian traditional spice which 

are widely used in the country. Following the recent efforts to look 

for healthy remedies with antimicrobial potential, the effects 

of essential oils of these medicinal plants were investigated on 

Salmonella typhimurium. 

Objectives: Chemical composition and antimicrobial effects of 

essential oils from plants were evaluated on Salmonella typhimurium 

and compared with five antibiotics enrofloxacin, chloramphenicle, 

erytromycin, oxytetracyclin and flumequin. 

Methods: The essential oils of the leavs and stems from each plants 

obtained by hydrodistillation by clevenger and analyzed by GC/MS. 

Results: The results of this study showed that Thyme oil more effective 

compared with flumequin, oxytetracyclin and erythromycin (P

multiple fracture (n=1, 1.3%). The animals were treated 

conservatively with a walking cast bandage (n=40, 55.5%), 

intramedullary pinning (n= 12, 16.6%) plate fixation (n= 1, 

1.3%)amputation (n=3, 4.1%). Sixteen owners (18%) did not appliy 

for a treatment. 

Conclusions: Accurate application must necessary for concerning 

fracture prophylaxis in newborn and young calves during parturition, 

especially proper practice of forced extraction. Succes of fracture 

healing depend on timing and proper selection of treatment. This 

study shows that intramedullary pinning fixation give a satisfactory 

results of calves with fracture. But, final decision of the 

intramedullary pinning is depend on patient condition and fracture 

types. 

Key words: calves, fracture, intramedullary pinning 

310 Evaluation of the Treatment of Septic Arthritis by 

Arthrotomy in Calves 

A. Sartelet, K. Touati 

University of Liege, Clinical Department of Food Animals, Liege, 

Belgium 

The objectives of this study are to evaluate the effectiveness of the 

treatment of chronic septic arthritis of the tarsus, stifle or elbow 

joints by arthrotomy in calves. This study included 29 Belgian blue 

calves suffering from chronic septic arthritis (CSA) of the elbow, 

the stifle or the tarsus referred to the Clinic for ruminants of Liege. 

For the arthrotomy, after an induction (xylazine), an intravenous 

general anaesthesia (triple drip) was used associated with a high 

epidural anaesthesia for the fore limbs. A single incision on the 

lateral side of the joint was performed except for the tarsal joint 

where a second incision on the medial side was also performed. The 

joint capsule and the cartilage were curetted. The joint was flushed 

with 2 l of chlorhexidine diluted solution. The incision was sutured 

except for the tarsus. Penicillin (20.000 UI/kg, IM, SID for 15 

days), gentamicine (6 mg/kg, IV, SID for 5 days), flunixine (2.2 

mg/kg, IV, SID) and cefalexine (5 ml, SID, intra-articular) were 

administered. Telephone follow up 3 months after surgery was 

performed. All the calves had been previously treated for 10-13 

days with antibiotics and anti-inflammatory drugs in farm, without 

success. Thirteen tarsal septic arthritis including 10 with signs of 

osteomyelitis, 11 of the stifle including 6 with osteomyelitis and 5 

of the elbow including 3 with osteomyelitis were diagnosed. Eight 

calves suffering from tarsal CSA were treated by arthrotomy and 5 

had a good surgical outcome. Five calves were euthanatized for 

economic reasons. Seven calves suffering from CSA of the stifle 

were treated by arthrotomy and the others were treated medically. 

Four had a good surgical outcome. Two calves suffering from CSA 

of the elbow were treated surgically with success, 1 was 

euthanatized and 2 were treated medically. CSA in calves represents 

25.2 % of the consultations for lameness in cattle. CSA of the stifle, 

elbow and tarsus represent 31 % of all the cases of arthritis. Eleven 

of the 17 (65 %) treated calves had a good surgical outcome. These 

animals were fattened or used for breeding. Arthrodesis being 

unrealizable on these joints, the arthrotomy seems to be a 

reasonable option for calves suffering from CSA when medical 

treatment and joint lavage were ineffective and given the poor 

prognosis of the disease. 

Key words: calves, lameness, arthritis, arthrotomy 

341 Chronic Septic Arthritis of the Carpus: Surgical Approach 

K. Touati, A. Sartelet 


Belgium 

The objective of this study was to evaluate the effectiveness of 

arthrodesis in surgical treatment of carpal septic chronic arthritis in 

calves. This study included Belgian Blue calves presented to the 

University of Liege for lameness. The diagnosis was based on 

general and orthopaedic exams and confirmed by radiography and/or 

ultrasonography. For arthrodesis, sedation with xylazine was 

performed followed by a loco regional intravenous anaesthesia with 

a tourniquet. A dorsal circular incision from the lateral to the medial 

side of the joint was performed. Tendons, ligaments and synovial 

membrane were incised. The 3 articular levels of the carpus were 

opened. All the cartilage and the damaged subchondral bone were 


curetted. The joint was flushed with a chlorhexidine diluted solution. 

A cannula was placed in the joint before suturing the skin with 

mattress sutures using a synthetic absorbable suture material. A cast 

including the elbow joint was performed. Penicillin, gentamicine, 

flunixine and cefalexine (5 ml, SID, intra-articular) were 

administered. The cast was removed 5 days postoperatively and the 

surgical site was checked. If evolution was good, a new cast was 

placed and the calf was discharged. The cast was changed again 3 

weeks after the first recheck and a new one was placed for another 3 

weeks. If evolution was not satisfactory, the decision to continue the 

treatment or not was taken with the farmer. Telephone follow up was 

performed 3 months after surgery. Septic chronic arthritis was 

diagnosed in 97 calves. The mean age SD was 2 1.2 months and the 

mean body weight SD was 74 ± 29 kg. Twenty nine septic chronic 

arthritis of the carpus, 21 of the fetlocks, 13 of the tarsus, 11 of the 

stifle, 5 of the elbow, 3 of the hip, 2 of the intervertebral joint, 1 of 

the distal interphalangeal joint, 1 epiphysitis and 11 polyarthritis 

were diagnosed. Twenty two arthrodesis of the carpus were 

performed of which 15 had a good surgical outcome. Ten calves 

were fattened and 5 were used for breeding. Arthritis of the carpus 

represented 30 % of all cases of arthritis in calves in this study. 

Seventy percent of the cases of carpal arthrodesis had a good 

surgical outcome. Arthrodesis is the best treatment for arthritis of the 

carpus when medical treatment and joint lavage are ineffective. 

Arthrodesis improves prognosis and decreases economic losses 

related to this disease. 

Key words: cattle, calves, arthritis, arthrodesis 

342 Inflammatory Response after Surgical Correction of Left 

Displaced Abomasum using Different Techniques 

T. Wittek 1 , A. Grosche 2 , M. Fürll 1 

1 University of Leipzig, Medizinische Tierklinik, Leipzig, Germany 

2 University of Florida, Department of Large Animal Clinical 

Sciences, Gainesville, United States 

Objective: Left displacement of the abomasum (LDA) is a 

frequently occurring condition in lactating dairy cows. Different 

techniques have been developed for surgical correction of LDA that 

do (abomasopexy) or do not (omentopexy) require perforation of the 

abomasal wall. The objective of this study was to compare the 

inflammatory response within the peritoneal cavity and in the leukon 

between three surgical methods. 

Material and Methods: The study comprised 45 lactating dairy cows 

with LDA which were alternately allocated into 3 groups (n=15). The 

surgical techniques were laparotomy and omentopexy in right flank 

(group RFO), left flank laparotomy and right ventral omentopexy 

(group LFO) and laparoscopic abomasopexy (group LA). Only the 

laparoscopic method requires perforation of the abomasal wall. A 

physical examination was performed daily. Blood and peritoneal fluid 

samples were obtained before and for 3 consecutive days after surgery. 

White and red blood cell count, and biochemical analysis (total protein, 

albumin, glucose, urea, creatinine, cholesterol, fibrinogen 

concentration; AST, CK, ALP, LDH activities) were measured in 

blood and peritoneal fluid. 

Results: Cows after laparoscopic abomasopexy had higher post 

operative food intake and increased rate of rumen contractions in 

comparison to cows undergoing laparotomy. Number of leukocytes 

in peritoneal fluid increased after surgery in all groups (RFO: from 

3.3 ± 1.8 to 17.4 ± 13.8 G/L; LFO: from 2.9 ± 1.2 to 14.7 ± 10.0 

U/L; LA: from 3.3 ± 2.0 to 38.9 ± 23.6 G/L). The leukocyte 

concentration in peritoneal fluid was significantly higher after 

laparoscopy than laparotomy. Serum CK activity increased 

significantly after surgery in cows after laparotomy (RFO: from 370 

± 228 to 633 ± 470 U/L; LFO: from 487 ± 294 to 752 ± 325 U/L) but 

not after laparoscopy (LA: from 432 ± 301 to 280 ± 187 U/L). CK 

and LDH activities in peritoneal fluid were also significantly 

increased only after laparotomy. 

Conclusions: Surgical correction of LDA by laparoscopic 

abomasopexy produced a faster rate of clinical convalescence (based 

on post operative food intake and rumen motility). Laparoscopy did not 

result in skeletal tissue damage, compared to laparotomy (based on 

changes in CK and LDH activity). The increased peritoneal fluid 

leukocyte concentration after laparoscopy is most likely due to 

perforation of the abomasal wall. 

Key words: left abomasal displacement, laparotomy, laparoscopy, 

peritonela fluid, inflammation

343 Effect of Displacement of the Abomasum on Performance in 

Dairy Cattle after Laparoscopic Correction 

R. Jorritsma 1 , B. Westerlaan 3 , M. Bierma 2 , K. Frankena 2 

1 Utrecht University, Farm Animal Health, Utrecht, The Netherlands 

2 Wageningen University and Research Centre, Quantitative 

Veterinary Epidemiology, Wageningen, The Netherlands 

3 Veterinary Practice Oosterwolde, Oosterwolde, The Netherlands 

The aim of the study was to investigate the effect of laparoscopically 

corrected left displacement of the abomasum (LDA) on milk 

production and survival in Holstein-Friesian dairy cattle. The study 

consisted of 91 cows diagnosed with LDA and 195 control cows 

matched for herd, parity, and calving date. At the time of surgery the 

cow’s parity and latest calving date were registered. Data on 

insemination, milk production and culling was collected from 

various sources. Ninety percent of the LDA treatments were 

performed within 4 weeks after calving. The risk of being culled 

during the total observational period of at least 3 year was 1.5 times 

as high for LDA cows compared to their matched herd mates (p = 

0.009). The risk of being culled in the current lactation was higher 

for LDA cows (hazard ratio 1.8, p = 0.013). However, risk of being 

culled after the next calving following LDA correction was similar 

for both groups. For the lactation in which LDA correction was 

performed, there was no difference in 305 day milk production 

between LDA (n = 80) and matched cows (n=182). However, mean 

calving to first service and mean calving interval during the same 

lactation was prolonged for LDA cows (115 vs 98 days and 451 vs 

418 days respectively). 

344 Generalised Peritonitis in Cows due to Perforation of an 

Abomasal Ulcer into the Abdominal Cavity 

A. Lorch, Ch. Heizer, T. Zauscher, G. Rademacher 

University of Munich, Clinic for Ruminants, Oberschleissheim, 

Germany 

History, clinical findings and course of disease as well as surgery 

and/or necropsy findings in 13 cows with a confirmed perforation of 

an abomasal ulcer into the free abdominal cavity are described. The 

data were collected in a prospective study involving 29 cows 

admitted to our clinic between August, 2004, and October, 2005, that 

fulfilled at least five out of six of the following selection criteria 

worked out in a prior retrospective study. 1) age over two years, 2) 

eight weeks post partum, 3) no or only mild signs of colic, 4) 

abnormal posture, 5) abnormal shape of the abdomen, 6) increased 

tension of the abdominal wall. 

All cows with a generalised peritonitis due to perforation of an 

abomasal ulcer into the abdominal cavity (n=13) were detected with 

these criteria. All patients except one were Simmental cattle. The 

majority of them was over four years old (69 %). 10 out of 13 had 

calved in the previous two weeks. Most of the cows had been sick 

two or three days before a sudden deterioration prior to admission. 

All cows showed a comparatively uniform clinical picture. The 

general condition was reduced in all patients, mostly severely so. All 

of them were severely dehydrated. The abdomen was distended in all 

cases and, as a significant clinical sign, the paralumbar fossae were 

symmetrically flattened to varying degrees (once even protruding). 

A so called double wall phenomenon was felt in the left paralumbar 

fossa in all 12 cases that were examined for this sign. The tension of 

the abdominal wall was increased in all cows. Percussion and 

swinging auscultation produced rather dull ping and splashing 

sounds, respectively, of low intensity. The liver could not be 

detected by percussion. Rectal examination showed a reduced or not 

existing vacuum in the abdomen. Pain provocation was positive in 

11 out of 13 cases. Many times ultrasonography and tap of the 

abdominal cavity added to the preliminary diagnosis. The main 

surgery and/or necropsy findings included free gas and increased 

abdominal fluids, greenish and moist masses of fibrin close to the 

abomasum. The perforation was always located in the corpus region 

of the abomasum. The main differential diagnosis in the other cows 

with fulfilled selection criteria was peritonitis of other origin (e.g. 

foreign body, perforated ulcer of the colon). The differences in 

location of the perforated abomasal ulcer and pathway of perforation 

in cows and calves are discussed. 

Key words: cattle, cow, generalised peritonitis, perforating abomasal 

ulcer 

345 Intestinal Intussusceptions in Cattle: a Retrospective Study 

(22 Cases) 

A. Sartelet, H. Guyot, S. Vandeputte, K. Touati 


Belgium 

The objective of this study is to determine the prevalence and the 

prognosis after surgical treatment of intestinal intussusceptions (I.I.) 

in cattle. This study included 22 Belgian blue cattle, 11 calves (C) 

and 11 adults (A) referred to the clinic for ruminants of the 

University of Liege on 14 months. A clinical exam, a biochemistry 

profile, a haematology profile and a trans-abdominal 

ultrasonography were performed. A celiotomy on dorsal recumbency 

on the midline under intravenous general anaesthesia for the calves 

and a right flank celiotomy on standing animal for the adults were 

performed. When manual reduction was inefficient, an enterectomy 

was performed. Fluidotherapy, Penicillin (IM, SID for 10 days), 

gentamicine (IV, SID for 5 to 10 days), flunixine (IV, SID for 4 

days) were administered post-operatively. In all cases, they were 

referred for decreased in appetite and scant fecal material. In all 

cases, abdominal distension was observed. In 10 cases (3 C/7 A), 

signs of abdominal pain were observed. Fecal material was scant in 

4 cattle (1 C/3 A), absent in 13 (7 C/6 A) and contained blood in 5 (3 

C/2 A). Biochemistry and haematology revealed a hypochloremic 

metabolic alkalosis in 16 (5 C/11 A) cases and acidosis in 5 calves. 

In 6 cases, ultrasonography allowed diagnosis. There were 13 

jejunojejunal (4 C/9 A), 4 ileocecal (3 C/1 A), 4 ileoileal (4 C/0 A) 

and 1 colocolic (0 C/1 A) intussusceptions. Sixteen (7 C/9 A) cattle 

were treated by enterectomy, 3 (1 C/2 A) by manual reduction, 2 

were euthanatized during the celiotomy (2 C/0 A) and one died 

before the surgery (1 C/0 A). Five calves died during the 

hospitalisation and 3 were euthanatized. Ten adults recovered well 

and were discharged 4 to 8 days after surgery. One adult was 

euthanatized. All necropsies revealed fibrinous peritonitis and 

complete intestinal ileus. Unlike other studies, our study showed that 

I.I. represented 20 % of the surgical intestinal obstructions during 

this period. The season had no impact in our study. 

Clinicopathological findings were not specific. Ultrasonography 

may be a useful diagnostic tool. No aetiology could be determined 

from this study. Similar to other studies, I.I. occurred most 

commonly (60%) in the jejunal portion. The overall success rate 

after surgery was 50 %. This study also revealed that I.I. has a better 

prognosis for adults (91 % success rate) than calves (9 % success 

rate). 

Key words: cattle, intestinal obstruction, intussusception, enterectomy 

346 Abomasal end-to-end Anastomosis as a Treatment for an 

Extensive Abomasal Hernia and Fistula in a Cow 

G. Vertenten 1 , J. Declercq 1 , A. Martens 1 , L. Devisscher 1 , 

S. Torfs 2 , G. Van Loon 2 , F. Gasthuys 1 


Surgery and Anaesthesiology of Domestic Animals, Merelbeke, 

Belgium 


Internal Medicine and Clinical Biology of Large Animals, 


A 37-month old Holstein-Friesian cow was referred to the clinic with 

a bleeding abdominal hernia. Eleven months earlier,a left 

paralumbar fossa omentopexy had been performed as treatment of a 

left displaced abomasum. The cow had recovered well and calved 

normally. A few days after calving, the omentopexy suture got 

infected and was removed. Afterwards a fleshy mass protruded 

through the skin where the suture removal had been performed. 

General examination revealed no signs of dehydration or anaemia. 

Potassium and calcium blood concentrations were low (2.3 and 

0.7mmol/l respectively) while venous base excess was high (26 

mmol/l). A large fleshy mass (15 cm diameter) with superficial 

necrosis and diffuse bleeding protruded through a hernial ring 

cranial to the umbilicus. An additional fistula was present towards a 

cavity filled with ingesta. The pH of the fluid was below 5, 

suggestive for an abomasal fistulation. Ultrasound of the hernia and 

protruding mass only revealed oedema with acoustic shadowing 

while abdominal ultrasound showed dilated small intestines. Surgery 

was performed in the anaesthetized cow in the dorsal recumbent 

Surgery, Anaesthesia, Pain Management and Diagnostic Imaging 257

position. A fusiform skin incision was created around the protruding 

mass and the hernial ring and abdominal wall were exposed. The 

hernial ring was excised and the protruding mass was identified as 

the abomasum. A large part of the abomasal body (including the 

curvatura major and minor) was involved in the hernia. After 

removal of all necrotic tissue, the abomasum was reconstructed 

using an end-to-end anastomosis. The surgical procedure was 

accompanied with a severe haemorrhage inducing a decrease of the 

packed cell volume from 24 to 17%. Recovery from anaesthesia was 

uneventful. An intensive intravenous supportive therapy was 

installed afterwards (lactated ringer solution supplemented with 

glucose, calcium and potassium chloride). Apart from some oedema 

around the abdominal wound, the cow had fully recovered after 10 

days and was dismissed from the clinic. The subsequent lactation 

was uneventful. 

Abomasal fistulation has rarely been described as a complication of 

omentopexy. This fistulation might be caused by inadvertent 

penetration of the abomasal lumen during a standard omentopexy 

procedure. Eventually, abomasal ulcerations can be induced by the 

chronic inflammation following the omentopexy, creating a 

fistulation through the abdominal wall. The combination of an 

abomasal fistula and an abdominal hernia can end in a complete 

herniation of the abomasum, requiring an end-to-end anastomosis. 

424 Laparoscopy-assisted Biopsy of Mesenteric Lymph Nodes in 

Calves 

T. Seeger1 , P. Berisha1 , K. Koehler2 , M. Buelte3 , K. Doll1 1 University of Giessen, Clinic of Ruminants, Giessen, Germany 

2 University of Giessen, Institute for Veterinary Pathology, Giessen, 

Germany 

3 University of Giessen, Institute of Veterinary Food Science, Giessen, 

Germany 

Objectives of study: Examination of mesenteric lymph nodes is still 

the “gold-standard” for diagnosis of infections with Mycobcterium 

avium ssp. paratuberculosis in cattle. Results of other studies have 

shown that this bacterium is also detectable in mesenteric lymph nodes 

of young animals. Aim of this study was to validate a laparoscopic 

method for the biopsy of these lymph nodes in calves. 

Material and Methods: The study was performed on 15 Holstein 

calves. All calves were between 2 and 4 weeks of age. During the 3 

week study period the animals were housed in the clinic. To identify 

complications after laparoscopic surgery clinical examinations were 

performed twice daily and blood samples were taken on several 

study days. At the end of the study period all calves were euthanised 

and a necropsy was performed at the Institute for Veterinary 

Pathology. The laparoscopy-assisted biopsy was performed on day 7 

of the study. Before surgery all calves were treated with antibiotics 

and antiphlogistics for one time. Detomidine (0.05 mg/kg im) and 

ketamine (6 mg/kg iv) were admistered for anesthesia. The calves 

were fixed on an operating table in right lateral recumbency. The left 

abdominal wall was prepared for aseptic surgery. Three stab 

incisions of the skin were performed and a Verres needle was 

inserted into the abdomen to create a pneumoperitoneum. Three 

magnetic valve trokars were inserted such that a rigid laparoscope 

and 2 forceps could be introduced into the abdominal cavity. After 

visualisation and fixation of the cecum the ileocecal lymph node was 

fixated and a biopsy was taken with a special spoon forceps. Same 

procedure was performed for biopsy of one lymph node adjacent to 

the caudal part of the jejunum. At the end of surgery the 3 skin 

incisions were closed by a simple suture. 

Results: No complications due to the anesthesia or the 

pneumoperitoneum were observed. The average operation time was 

26.2 minutes with a range from 18 to 37 minutes. The biopsy of a 

jejunal lymph node was successful in all calves. In one calf only the 

biopsy of the ileocecal lymph node was impossible. No major 

postoperative complications were observed. During the necropsies no 

signs of peritonitis or wound complications were detected. 

Conclusions: The results of this study show that the described 

laparoscopic method for biopsy of mesenteric lymph nodes in calves is 

well suited to obtain these important specimens for paratuberculosis 

diagnosis. 

Key words: laparoscopy, lymph node biopsy, paratuberculosis, calf 

425 Two-dimensional and M-mode Echocardiographic 

Parameters Measured in Fifteen Healthy Calves 


N. Cesbron 1 , V. Charot 2 , A. Dorizon 2 

1 National Veterinary School of Nantes, Unité de Médecine des 

Animaux d’Elevage, Nantes, France 

2 National Veterinary School of Nantes, Unité d’Imagerie Médicale, 

nantes, France 

Objectives of study: Within ultrasonography procedures, 

echocardiography has not been studied as extensively in cattle as in 

pets. Among all the studies dealing with echocardiography in cattle, 

most of them were mainly focused either (i) on examinations of cardiac 

affections (like endocarditis, pericarditis or congenital abnormalities) 

or (ii) in healthy adult cattle. Few studies have been made in calves. 

Therefore, the aim of this study was to obtain reference values for the 

echocardiographic parameters in healthy calves using both twodimensional 

real-time and M-Mode echography. 

Materials and methods: To reach this goal, fifteen Holstein 

clinically healthy calves were used. Their ages ranged from 2 weeks 

to 1 month. The examinations were made while the calves were 

standing using a 2.5 MHz convex transducer. The heart was 

examined ultrasonographically on the right and then the left side 

with the transducer in four positions: caudal long, caudal short and 

cranial long axes on the right side; caudal long axe on the left side. 

The variables were measured in a short axis view of the heart and 

both two-dimensional (2D) and cursor- directed time-motion (M) 

modes were used. Measurements, at systole (s) and diastole (d), of 

the left ventricular internal diameter (respectively LVDs and LVDd), 

and of the interventricular septum thickness (respectively IVSs and 

IVSd) were obtained according to the previous described procedure. 

The left atrial diameter (LAD), the aortic root diameter (AOD) and 

their ratio [LAD/AO] were described. 

Results: For the ventricular internal diameter at systole and diastole, 

the following results (in mm) were obtained with respectively [LVDd: 

44.57 ± 6.53], [LVDs: 30.07 ± 5.82]. For the interventricular septum 

thickness, we observed at systole and diastole (in mm) [LVSd: 11.90 ± 

2.33], [IVSs: 16.64 ± 2.19]. Lastly, concerning the left atrial and the 

aortic diameters, we observed (in mm) [AOD: 31.18 ± 3.02], [LAD: 

34.31 ± 4.26] leading to a ratio [LAD/AO: 1.11 ± 0.17].These results 

are slightly lower than those reported by a previous study performed by 

Amory et al. on older calves (> 2 months). 

Conclusion: The echocardiographic reference values obtained on these 

healthy calves aged from less than one month were similar to those 

classically reported for dogs of similar body weight (large breed size). 

Key words: echocardiography, calves, M-mode and two-dimensional 

echocardiography 

426 Measurements of Dimensions of Holstein-Friesian Calves 

Using Computed Tomography in order to Compare Different 

Obstetrical Traction Methods 

C. Heun 1 , G. Tsousis 1 , M. Becker 1 , J. Rieder 2 , H. Bollwein 1 

1 University of Veterinary Medicine Hannover, Clinic for Cattle, 


2 University of Veterinary Medicine Hannover, Small Animal Clinic, 


The aim of this study was to measure the difference in calves’ 

dimensions resulting from different obstetrical methods. 20 stillborn 

calves (41.1 +/- 3.7 kg) were measured objectively by computed 

tomography. The calves were placed stretched on their right side 

simulating the simultaneous traction with both front legs on the same 

level (0D), the alternate traction with 5 cm difference between the level 

of the legs (5D) and the alternate traction with 10 cm difference 

between the level of the legs (10D). All measurements were carried out 

with special software. For the statistical analysis repeated measures 

ANOVA was used. For each traction the maximum cross-sectional area 

on the level of the forehead (ma-F) and on the elbows, the biggest width 

of the shoulders on the level of the tuberculum majus (width-tm) and 

the perimeter on the level of the forehead (peri-F) were measured. The 

ma-F reduced from traction 0D to 5D with a mean difference of 2.9 ± 

4.1 cm (p=0.005), from 5D to 10D with 8.8 ± 5.8 cm (p

(p

Conclusions: It is the first presentation of clinical results of half 

circular external fixator by a new, own conception, which was used 

in sheep for the treatment of tibia fractures, and one of the 

conclusions are that this apparatus can be effectively and safely 

employed in sheep. However, due to a small number of clinical cases 

concerning treatment of the long bone fracture with these halfcircular 

rings external fixators, further studies are needed. 

959 Management of Left Abomasal Displacement in Iraqi Buffalo 

M. Al-Badrany 

College of Veterinary Medicine, Mosul University, Department of 

Surgery, Mosul, Iraq 

Twenty five cases of abomasal displacement in Iraqi buffaloes were 

examined in the privately own veterinary clincof the author during 

the period between February 2006 to June 2007). Corrective surgery 

was performed, and 98% of these recovered completely. Surgery 

was, in most cases, performed on the same day as the diagnosis of 

abomasal displacement was made. The displacements occurred at all 

times of the year: 86% of cases involved buffaloes in third or 

subsequent lactation 95% were diagnosed within the period 4-64 

days after calving, and 70% of the buffaloes were treated for other 

illness with disturbance of digestion during the week immediately 

prior to the demonstration of the displacement. Further, 25% ketosis 

was diagnosed at the same time with the displacement. Etiology and 

pathogenesis are discussed 

960 Management of Some Bovine Claw Affections by Honey and 

Some Medicinal Plant Extracts 

M. Ali 

Faculty of Veterinary Medicine, Assiut University, Veterinary 

Surgery, Assiut, Egypt 

The investigation conducted 93 cows suffering from different claw 

affections out of 965 lactating Friesian cows. Sole ulcers, interdigital 

dermatitis, interdigital mecrobacillosis and digital dermatitis 

resempled 16.1, 32.2, 19.1 and 33.3 %, respectively. Aerobic 

baterial aetiology (101 isollates) was Staph. aureus, Strept. pyogens 

and Corynebacterium pyogens as 37.6, 1109 and 50.5 % 

respectively. Anaerobic ones (11.5 isolates) were Fusobacterium 

necrophorum, Bacteroides nododus, Peptostreptococcus anarobius 

and Clostredium sp. resmpling 62.6, 19.1, 14.8 and 3.5 % 

respectively. Antimicrobial resistance assessment revealed that all 

isolated bacterial strains were multidrug resistant where 174 of them 

(80.5%) Showed quintuple resistance pattern. Ten pencentage of 

Thymus vulgaris extract inhibited all isolated bacteria, while both 

Marticaria chamomella and Origanum vulgare showed minimum 

inhibitory concentrations as 10 and 15% for aerobic and anaerobic 

bacterial isolates respectively. The different claw affections were 

managed separately using four different materials; lotion and 

ointments of the medicinal plant extracts mentioned above as well as 

honey as it is. The achieved healing processes were judged by foot 

scores (pain detection, lameness and lesion dimensions retraction). It 

was concluded that alternative medical approach is highly 

recommended especially when the aetiology is multidrug resistant 

and the obtained results declared the superiority of honey use 

followed by Thymus vulgaris, Marticaria chamomella and 

Origanum vulgare, respectively. 

Key words: management, bovine claw, honey 

961 Foreign Body Syndrome in Cattle of Rajasthan, India 

T. Gahlot, N. Arora 

College of Veterinary and Animal Science, Veterinary Surgery & 

Radiology, Bikaner, India 

The present report is based upon a clinical study of 50 cows of age 

group from 3-8 years suffering from foreign body syndrome. These 

animals were dull and depressed and had depraved appetite or 

complete inappetance, tympany of mild to severe nature, impaction 

of rumen and scanty and mucous stained faeces. The temperature, 

pulse and respiration of these animals were in normal range except in 

state of tympany. The ruminal movements were decreased and 

reticular sounds were either absent or less than normal. In cases of 

impaction the left flank appeared hard. The aspiration of ruminal 


fluid by a percutaneous paracentesis of rumen revealed pH to be 

between 7.5-9. The left flank laparorumenotomy was performed 

under local infiltration anaesthesia using 2% lidocaine 

hydrochloride. In cases of impaction of rumen the ruminal contents 

were in form of enmeshed single mass composed of polythene bags, 

ropes, leather shoes, rags, etc. There was sand, pebbles, iron 

particles, glass pieces and bones as sediment in rumen. The thorough 

searching of reticulum revealed potential (wire, needle, nail etc) and 

non potential (nut, bolt etc) foreign body. In other cases, few non 

potential foreign bodies composed of rags, polythene ropes, sand 

pebbles etc were recovered from the rumen and 2-5 potential foreign 

bodies composed of broken iron pieces, nails, wires, etc were 

recovered from reticulum. In these cases the rumen liquor was 

present in adequate quantities. However, in 12 of these cases 

adhesions of reticulum with peritoneum were found which was 

suggestive of the fact that foreign body migration has occurred. The 

rumen pH was adjusted by administering vinegar or acetic acid into 

the rumen. In cases where rumen cud or liquor was either absent or 

scarce the additional 5 litre of fresh rumen liquor recovered from 

freshly slaughtered cattle was administered orally for 3 days. In 

cases of impaction animals started rumination after 1 week but in 

other cases the rumination was restored on 3 rd -4 th day post 

operatively. All these cases were supplemented with a mineral and 

vitamin mixture in the concentrate and out of 50 cows having 

impaction 3 died after 3-5 weeks of operation. The post mortem of 

these animals revealed severe purulent peritonitis while the cases of 

impaction had a simultaneous impaction of omasum and ulceration 

at abomasal mucosa. 

Key words: cow, foreign body syndrome, India 

962 First Field Experiences with the Use of a Monofilament 

Synthetic Absorbable Suture Material (Monoswift ® ) for 

Uterine Surgery 

J. Coghe 

Veterinary Practice VDN, Roeselare, Belgium 

Objectives of study: A monofilament synthetic absorbable suture 

(Monoswift ® EP5/USP2, CP Medical) was used during caesarean 

sections in cattle for uterine closure. A follow-up of complications 

during and after surgery as well as fertility was done. 

Materials and methods: Under field conditions, 42 Belgian White and 

Blue cattle (heifers n=28, 2 nd parity n=13, 3 rd parity n=1) had a 

caesarean section done by one veterinarian. The uterus was closed with 

Monoswift ® by a double suture: a first perforating continuous suture 

followed by a cushing type suture. Complications during and after 

surgery were registered. A rectal examination was performed on 15 

cows one and two months after surgery. Finally, a follow-up was done 

of fertility by checking on the number of cows that were pregnant 

afterwards and by checking their calving interval. 

Results: Manipulation of the monofilament was excellent and slight 

elasticity assured tightness of the suture when the uterus regresses. 

Retentio secundinarum happened in 14% of the cows and they were 

treated with antibiotics. None of the cows showed signs of peritonitis 

after surgery. Rectal examination one month after surgery permitted to 

palpate the knot on uterine surface in 33% of the cows. No suture 

material was palpated two months after surgery. Two cows had a small 

local adhesion of the uterus at rectal examination two months after 

surgery. Thirty-five cows were inseminated afterwards and 91% were 

confirmed pregnant. Four of them had an abortion. Mean calving 

interval was 387.6 ± 45.9 days. 

Conclusions: Caesarean sections are performed daily by veterinary 

practitioners. Uterine suture is definitely the key factor in a good 

outcome of this surgery. Therefore, a good surgical technique in 

combination with the best surgical material is required. This 

experiment in field conditions showed that Monoswift ® possesses all 

the characteristics for a tight uterine suture without any negative effect 

on fertility afterwards. Absorption is faster than other monofilament 

synthetic absorbable sutures and makes this material the preferred one 

for uterine suture. 

Key words: uterine suture - monofilament 

963 Effects of Mephentermine and Norepinephrine 

Administration Following Thiopentone Induced Circulatory 

Insufficiency in Buffaloes 

P. Peshin, J. Singh, R. Tyagi, A. Kumar, S. Singh

Chaudhary Charan Singh Haryana Agricultural University, College 

of Veterinary Sciences, Veterinary Surgery and Radiology, Hisar, 

India 

The study was conducted in ten buffalo calves, 1-2 years old, 

weighing 65 to 120 kg, randomly divided into two groups of five 

animals each. Circulatory insufficiency (CI) was induced with 

thiopentone sodium (5%), and then in one group I, mephentermine 

(0.5 mg/kg, I.V.) and in group II, norepinephrine (2.0 mg 

norepinephrine in 500 ml normal saline solution equivalent to 4.0 

µg/ml, I.V. infusion) was administered for treatment. Various 

parameters investigated were: rectal temperature (RT), haemoglobin 

(Hb), plasma glucose, plasma creatinine, total plasma proteins, heart 

rate (HR), systolic pressure (SP), diastolic pressure (DP), mean 

arterial pressure (MAP), central venous pressure (CVP), 

electrocardiogram(ECG) electroencephalogram (EEG), arterial 

carbon dioxide tension (PaCO 2), arterial oxygen tension (PaO 2) and 

venous oxygen tension (PvO 2).Restoration of corneal, palpebral 

reflexes and papillary reflex began within 15 minutes of 

mephentermine administration. The MAP was significantly elevated 

(21.06±1.191 kPa) five minutes after mephentermine as compared to 

5.99±0.794 kPa after CI and remained in the normal range 

(21.50±0.571 kPa). Following mephentermine administration highly 

elevated CVP (3.26±0.314kPa) was recorded during CI fell below 

the base line up to 15 minutes (0.76±0.027 kPa) and thereafter, 

remained in the normal range. Arterial O2 tension remained elevated 

(22.53±1.402 kPa) throughout the observation period. In the later 

stages of the experiment, there was mild decrease in the RT 

(36.7±0.29°C). Moderately elevated blood glucose levels during 

circulatory insufficiency (5.08±0.48 mmol/l) was further elevated by 

mephentermine administration (6.27±0.497 mmol/l). Corneal and 

palpebral reflexes started returning by 45 to 60 minutes after the start 

of infusion of norepinephrine. Cardiovascular action of the 

norepinephrine started 1.19±0.28 minute after its initiation and near 

normal range of the arterial pressure could be achieved by 4.81±0.94 

minutes (15.27±0.853 kPa). The elevated CVP (3.43±0.495 kPa) 

during CI also decreased (2.04±0.197 kPa) after start of 

norepinephrine, but effect was slow as values remained significantly 

elevated than base line up to 15 minutes. Arterial O 2 tension 

remained elevated. The study revealed that mephentermine 

controlled the thiopentone induced CI effectively. 

Key words: mephentermine, norepinephrine, buffaloes calves 

964 Normal Cross - Section Anatomy of the Bovine Digit: 

Comparison of Computed Tomography and Limb Anatomy 

H. Mohammadi 1 , A. Raji 1 , K. Sardari 1 , M. Heidarpour Bami 1 , 

B. Khoramian 1 , Z. Saberi 1 , M. Rasekh 1 , M. Mazandarani 1 , 

E. Shahroozian 1 , H. Staji 1 

1 University of Tehran, Clinical Science, Tehran, Iran 

2 Ferdowsi University of Mashhad, Faculty of Veterinary Medicine, 

Mashhad, Iran 

3 Ferdowsi University of Mashhad, Faculty of Veterinary Medicine, 

Clinical Science, Mashhad, Iran 

Diagnosis and treatment of digital lesion in dairy cattle is important 

because of ample financial injuries. The bovine digit is a complex 

structure with many joints, ligament and tendons. Radiography (R) 

and Utrasonography (U) have a limited value in the evaluation of 

soft tissue. Computed tomography (CT) in small animal, is 

frequently used to evaluate the foot. The purpose of this research is 

provide a collection of normal cross-section anatomy of the bovine 

digit using by CT. CT was conducted on two hind limbs, taken from 

normal adult cows. CT was peformed using Siemens (ARTX MOD 

E1) at 50 mA and 130kv. The limbs were presented by freezing for 

several days at -18°C then the limbs sectioned in three planets 

transverse, sagital and dorsoplantar in 1- cm slices with an electric 

handsow. The average CT- normal of the digits was 103. Identifiable 

anatomic structure was labeled on the line drawing of the limb 

sections and on the corresponding CT images. Images produced 

good delineation between cortex, medulla, corium and wall of the 

hoof. On CT image bone appeared hyper dense while tendons, 

ligament and wall of the hoof had similar shades of grey. Synovial 

fluid and blood vessels were hypo-dense. The dewclaws had a mixed 

opacity below the proximal phalanx. Average diameters of the deep 

digital flexor, superficial digital flexor, proximal sesamoidal bone, 

distal sesamoidal bone and phalanx were measured in all CT images. 

Since CT images are able to distinguish structures well, therefore it 

can use in distinction and effective treatment method of digit lesion 

in cow and the other animals. 

965 Affections of the Salivary Ducts in Buffaloes 

N. Misk 1 , M. Semieka 1 , T. Misk 2 , A. Ahmed 1 

1 Faculty of Veterinary Medicine, Assiut University, Veterinary 


2 Alexandria University, Veterinary Surgery, Alexandria, Egypt 

The aim of the present study was to determine different affections of 

the salivary ducts in buffaloes with special reference to diagnosis 

and treatment. The present work was carried out on a total number of 

35 buffaloes suffering form different affections of the salivary ducts. 

The majority of cases were collected during the periodical visits to 

veterinary clinics in 168 villages belong to 14 provinces in Egypt 

during the period from 1994 to 2007. Some cases were recorded in 

the veterinary teaching hospital of the faculty of veterinary medicine 

Assiut University. The recorded affections of the salivary ducts in 

buffaloes include; ectasia of the parotid duct (18), parotid duct 

fistula (14) and sialocele (3). Each case was subjected to full study 

including case history, clinical examination, diagnosis and 

treatment, whenever possible. Exploratory puncture and radiography 

were used for confirmation of diagnosis in some cases. Intraoral 

marsupialization was performed for treatment of parotid duct ectasia 

with highly encouraging results. Salivary fistula was corrected by 

one of two successful techniques; the first by reconstruction of the 

parotid duct using polyethylene tube to preserve the function of the 

gland and the second by ligation of the parotid duct just caudal to the 

fistual opening with nullifying the glands function. Sialoceles were 

corrected by removal of the mandibular salivary gland of the 

affected side in two animals only. The third animal was not operated 

by owner’s request. Successful results were obtained in one case 

only. The other case failed due to wrong side extirpation of the 

gland. In conclusion we can state that salivary duct affections in 

buffaloes were ectasia of the parotid duct, salivary fistula and 

sialocele. Treatment was available with successful results and good 

prognosis in ectasia of the parotid duct and salivary fistula and 

guarded in sialoceles. 

966 Body Surface Cysts in Some Farm Animals 

N. Misk 1 , T. Misk 2 , M. Semieka 1 , A. Ahmed 1 



2 Faculty of Veterinary Medicine Alexandria University, Veterinary 

Surgery, Alexandria, Egypt 

The aim of the present study was to determine different kinds of 

cysts present on the body surface of some farm animals with special 

reference to diagnosis and treatment. The present work was carried 

out on a total number of 94 animals of different species 

(Buffaloes=5, Cattle=18, Sheep=5, Goat=34, Camel=13, Horse=5 

and Donkey=14) suffering from different kinds of cysts on the body 

surface. All cases were collected during periodical visits to 

veterinary clinics in 168 villages at 14 provinces in Egypt during the 

period from 1994 to 2007. Diagnosis was established depending on 

case history, location, clinical signs, exploratory puncture and 

histopathological examination of excised cysts. Different kinds of 

cysts on the body surface were diagnosed namely; Dermoid (36), 

Branchial (7), Dentigerous (2), Tassel or Wattle (32), Thyroid (15) 

and Interdigital (2). Aspiration and injection of medicaments were 

performed for treatment of thyroid cysts; incision and evacuation 

were applied for treatment of dermoid cysts while surgical excision 

was done for many cysts such as branchial, dentigerous, tassel, 

interdigital and some dermoids. The results of treatment were 

encouraging without postoperative complications. In conclusion we 

can state that different varieties of cysts were observed on body 

surface in some farm animals and nearly all of them can be managed. 

The work was supplied by many colored photographs. 

Key words: cysts, body surface 

967 Predilection Seats of Body Surface Abscesses in Relation to 

the Way of Infection in Some Domestic Animals 

N. Misk 1 , M. Semieka 1 , T. Misk 2 






The aim of the present study was to determine the most common 

predilection seats of body surface abscesses in relation to the way of 

entrance of infection in some domestic animals. The present study 

was carried out on a total number of 469 animals (Buffaloes=81, 

Cattle=140, Sheep=141, Goats=48, Donkeys=36, Horses=9 and 

Camel=14) suffering from presence of abscesses at different 

localities on body surface. All cases were managed during the 

periodical visits to veterinary clinics in 168 villages at 14 provinces 

in Egypt during the period from 1994 to 2007. Diagnosis was 

established depending on case history, clinical signs and exploratory 

puncture. Surgical treatment was performed in all cases. The 

predilection seats of the abscesses were recorded. Results indicated 

that abscesses were detected at the parotid region (77), mandibular 

region (75), subconjunctival (19), cheek region (27), ears (5), neck 

(28), thoracic limbs (43), thoracic wall behind the elbow (18), 

ventral abdominal wall (35), umbilicus (39), pelvic limbs (60), 

gluteal region (24), perineal region (7), tail (9) and vulval lips (3). 

The predilection seats of abscesses were compared with the possible 

way of entrance of infection in all cases. Results indicated that the 

ways of entrance of infection may be; breach on the surface of the 

skin or mucous membranes, migrating infected foreign bodies from 

the digestive tract, using of non-sterilized needle for intramuscular 

injection, punctured or penetrating wounds by infected foreign 

bodies and blood and lymphatic borne infection in the course of 

some infectious diseases. 

Key words: abscess; body surface 

968 Hernias in Some Farm Animals 

N. Misk 1 , T. Misk 2 , M. Semieka 1 



2 Faculty of Veterinary medicine Alexandria University, Veterinary 


The aim of the present study was to determine different types of 

hernias affecting some farm animals with special reference to 

diagnosis and possible surgical treatment. The present work was 

carried on 566 animals (Buffaloes=214, Cattle=136, Sheep=99, 

Goats=47, Donkeys=58 and Horses=12) suffering from different 

types of hernias (umbilical = 149, abdominal= 209, inguinal=7, 

scrotal=12 and diaphragmatic = 189). Cases were recorded during 

periodical visits to veterinary clinics of 168 villages belong to 14 

provinces in Egypt during the period from 1994 to 2007. Results 

revealed that the incidence of different types of hernias in different 

domestic animals were highly variable. The most common hernia 

encountered in buffaloes was diaphragmatic hernia (173 out of 214). 

In the contrary umbilical hernias were recorded in small number in 

buffaloes (37 out of 214). In cattle the most common type of hernias 

was umbilical hernia (92 out of 136) while diaphragmatic hernias 

were recorded in a very small number as compared to buffaloes (16 

out of 136). In donkeys the most common hernias were abdominal 

(52 out of 58) and the same in sheep and goat the abdominal hernias 

were more common than other types (84 out of 99 and 37 out of 47 

respectively). In horses a little number of abdominal and umbilical 

hernias was recorded. Diagnosis was established depending on case 

history, clinical examination, radiography and exploratory puncture. 

All types of hernias were corrected surgically with successful results 

except animals suffering from diaphragmatic hernias which 

considered unreliable for surgical treatment. 

969 Non-septic Chemical Peritonitis in a Calf 

A. Meimandi, A. Raayat Jahromi 

School of Veterinary Medicine, Shiraz University, Shiraz, Iran 

Peritonitis is a regional or total inflammation of visceral and parietal 

surfaces of abdominal cavity. Primary response of peritoneum to 

stimulations is transudate secretion. A male three months Holstein 

calf was referred to the veterinary teaching hospital of School of 

Veterinary Medicine of Shiraz University. According to the owner 

statements, the calf was oliguric and umbilical region had swelled 


with little secretions since one month ago. In clinical observations; 

heart rate, respiratory rate and rectal temperature were normal and 

umbilicus was too rigid with a palpable mass but no secretion. 

Followed premedication with xylazine and local anesthesia, a 

midline incision was made on umbilicus. Subcutaneous tissues were 

fibrous and discolored to dark brown, which were removed 

completely. In order to more exploration, after opening of abdominal 

cavity, a lot of cream color fluid with a moderate odor was exited. In 

exploration of abdominal cavity, all visceral organs including 

intestines, omentum, abomasums, liver and mesentery were adhered 

together, so that their dissociation was impossible and all were 

covered by a fibrous layer. It seems that the calf has had a patent 

urachus from his birth, so that urine had been leaked to abdominal 

cavity, too, and it has been closed later. Urine was caused a chronic, 

diffused, non-septic (sterile) chemical peritonitis. 

Key words: calf, peritonotis, non-septic 

970 Clinical and Radiographic Evaluation of the Cefquinome 

Efficiency in the Treatment of Cows with Severely Footrot 

Associated to Septic Arthritis in Distal Interphalangeal Joint 

C. Anhesini1 , R. Tiveron3 , A. Silva2 , R. Vinne4 , C. Rodrigues1,2 1 Faculty of Veterinary Medicine and Zootechnic - UNESP, Surgery 

and Anaesthesiology Veterinary, Botucatu, Brazil 

2 School of Veterinary Medicine - UNESP, Clinic, Surgery and 

Animal Reproduction, Aracatuba, Brazil 

3 Veterinary Practitioner, Large Animal Clinic, Aracatuba, Brazil 

4 Intervet of Brazil Veterinary, Large Animal Clinic, Sao Paulo, 

Brazil 

The footrot and septic arthritis are high prevalence diseases in the 

bovine locomotor system, causing great economical losses. The footrot 

is an acute or sub acute necrotic infection in the skin of the claw, 

causing intense lameness and abrupt fall in the production. The septic 

arthritis of distal interphalangeal joint is the main sequel in the cases of 

severe foot diseases. This is an inflammation of the synovial membrane 

and surfaces as a result of the infectious process, characterized by 

lameness, local pain, heat and articular edema. Digital amputation is 

often used large infective process. However this method results in 

premature withdrawal, mutilation and animal devaluation. The 

antibiotic therapy requests application of susceptibility principles 

pharmacokinetics and pharmacodynamics of these therapeutic agents. 

The objective of this study was to evaluate the efficiency of the 

cefquinome, associated to local surgical procedures, in the treatment of 

cow with severe footrot and septic arthritis of distal interphalangeal 

joint. 10 dairy cattle were used in this study with indication of digit 

amputation due severe footrot and septic arthritis in distal 

interphalangeal joint. The animals were treated during 21 days with 1 

mg/kg sid of cefquinome (Cobactan ® ) subcutaneous and local 

treatment with trimming and curettage on the infected digit. The 

efficiency of the therapy was clinically evaluated before, during and 

after treatment through pictures, lameness degree and analysis of the 

characteristics of joint aspects and healing by radiographic exam. 

Every animals presented significant improvement in locomotion score, 

lesion signals and radiographic aspect of the joint, after the period of 21 

days of treatment. The radiographic exam after treatment showed 

absence of infective characters in the treated joint. In conclusion 

cefquinome associated to surgical treatment from cows with severe 

footrot and septic arthritis of distal interphalangeal joint presented 

excellent improvement of clinical status allowing animals to return the 

productive activity without digit amputation. This study demonstrated 

the possibility of success using a preservative treatment in cattle that 

already suffered other types of interventions without success. This way 

cefquinome (Cobactan ® ) was a valuable antibiotic therapy in infectious 

disease from bovine distal limb. 

Key words: footrot, cefquinome, dairy cattle 

971 Subcutaneous Bursitis in Buffaloes and Cattle 

N. Misk 1 , M. Semieka 1 , T. Misk 2 





The aim of the present work was to study different varieties of 

subcutaneous bursitis with special reference to their location,

diagnosis and possible treatments. Bursitis can be true when 

inflammation develops in congenital bursae and acquired when 

affects subcutaneous bursae. Subcutaneous bursitis is a common 

condition seen in buffaloes and cattle. The present study was carried 

out on 166 buffaloes and cattle affected with different forms of 

subcutaneous bursitis. All cases were examined during periodical 

visits to veterinary clinics of 168 villages belong to 14 provinces in 

Egypt during the period from 1994 to 2007. Species, sex, age and 

clinical signs for each case were recorded. Exploratory puncture was 

preformed for confirmation of diagnosis. Medical treatment was 

advised for cases of precarpal bursitis in standing position while 

surgical excision was recommended for peristernal and olecranon 

bursitis in recumbent position. Results revealed that olecranon 

bursitis was diagnosed in 63 buffaloes and 3 cattle. All animals were 

female adult at age group between 2-13 years. In the contrary, 

precarpal bursitis was recorded in 24 cattle and 6 buffaloes. All were 

adult female animals with age group ranged between 2-9 years. 

Presternal bursitis was diagnosed in 60 buffalo calves and 10 cattle 

calves and affected male more than females. All were young calves 

of age group between 2 months up to two years. The clinical signs of 

subcutaneous bursitis in all forms were highly diagnostic and 

exploratory puncture was highly confirmative. Surgery appeared to 

be the ideal treatment for cases of presternal and olecranon bursitis 

while medical treatment of precarpal bursitis by evacuation of the 

contents and several injections of anti- inflammatory drugs were 

more practical. In conclusion we can state that three forms of 

subcutaneous bursitis were diagnosed in buffaloes and cattle 

namely; olecranon, presternal, and precarbal. Subcutaneous bursitis 

can be corrected medically and surgically with encouraging results. 

972 Effect of Caustic Paste Disbudding on Cortisol and Behaviour 

of Dairy Calves Treated with Regional Anaesthetic 

G. Stilwell1 , R. Campos de Carvalho2 , M. Lima1 , D. Broom3 1 Faculdade de Medicina Veterinaria-Universidade Tecnica, Centro 

de Investigaçio Interdisciplinar em Sanidade Animal (CIISA), 

Lisboa, Portugal 

2 VetMondego, Departamento de Clinica, Montemor-o-velho, 

Portugal 

3 University of Cambridge, Centre for Animal Welfare and 

Anthrozoology, Cambridge, United Kingdom 

Caustic paste (sodium and calcium hydroxide) is a disbudding 

method frequently used on young dairy female calves. Very few 

studies have looked at the pain caused by chemical burns in cattle. 

The present study used plasma cortisol and four pain-related 

behaviours (head shaking, ear flicking, head rubbing, transition from 

lying) to assess pain for the first 50 minutes after caustic paste 

disbudding. Regional anaesthesia was done on the cornual nerve 

mid-way between eye and base of horn bud. Thirty five female 

Holstein calves (mean age 22 ± 4 days) were randomly allocated to 

four groups: 1) PD Caustic paste disbudded 5 min. after saline 

injection (n=7); 2) PDA Paste disbudded 5 min. after 2% lidocaine 

injection (n=10); 3) PDAF Paste disbudded 5 minutes after iv 

injection of 3 ml of flunixin-meglumine (1.4 mg/Kg) and 2% 

lidocaine (n=10); 4) SD: sham disbudded 5 min. after saline 

injection (n=8). Blood was collected at 5 min. before (base-line 

values) and 10, 30 and 50 minutes after disbudding. Behaviour was 

recorded for 10 min periods, just before each blood sampling. At 30 

and 50 minutes after disbudding the PD group showed a higher 

cortisol level compared to base-line (p=0.028) and to all other 

groups (p

of 101.55±4.94 mmHg. Afterwards the mean arterial pressure 

decreased. There was significant increase in the heart rate (80±3 per 

minute) at 2 minute of diazepam administration when compared with 

the base value of 57 ± 2 beats per minute. There was no appreciable 

change in the values of CVP at any stage of observation. The 

findings revealed that diazepam is a good muscle relaxant with 

minimal cardiopulmonary effects. 

Key words: Diazepam, cow calf, heart rate, MAP, CVP 

975 Surgical Removal of an Intraocular Malignant Melanoma in a 

Romanian Tsigai Ewe 

F. Beteg, A. Muste, N. Mates, L. Oana 

University of Agricultural Sciences and Veterinary Medicine, 

Department Surgery, Cluj-Napoca, Romania 

Melanomas are common in dogs, cats, and horses. They are observed 

occasionally as congenital defects in pigs, goats and calves. 

Melanomas usually are cutaneous neoplasms but can occur wherever 

melanocyte clusters are found. Eye and mouth are common sites, but 

meninges, bone, aorta, etc. are all reported sites. Although tumors 

within the globe may or may not be visible, their effect on ocular 

function can be dramatic, with visual impairments, discoloration, 

discomfort, or a shape changing of the ocular globe and 

exophthalmia. The globe tumors may be small and are often not 

apparent by the external exams of the enucleated globe. This paper 

describes the clinical, surgical and histopathological aspects 

of intraocular malignant melanoma on a Romanian Tsigai ewe. By 

our information and knowledge , intraocular melanoma has not been 

previously reported in a Romanian Tsigai.Tumoral growing was 

observed protruding out of the left eye at a two years old ewe of 

Romanian Tsigai. Clinical examination of the tumor was found to be 

partial infected and to cover the entire ocular surface. The eye was 

enucleated, and sampled to histopathological exams, and diagnosed 

as an intraocular malignant melanoma. The surgical removal 

by lateral enucleation was successful, because local excision was not 

amenable, with no sign of reoccurrence or metastasis nine months 

after surgical treatment. The success of the treatment is supposed to 

be due to the total removal of the tumor before to the development of 

metastasis. 

Key words: intraocular, malignant, melanoma, tumor 

976 Hemodynamic and Hemogasometric Effects of Subarachnoid 

Administration of Ropivacaine in Sheep undergoing 

Laparotomy 

R. Godoy, L. Gouvea, C. Leite, R. Almeida, FH. Ximenes, 

L. Gontijo, C. Pereira, R. Ferreira II, JR. Borges, A. Teixeira Neto 

Universidade de Brasília, FAV, Brasília-DF Brazi 

Objectives of study: The objective of the present study was to evaluate 

the sensitive and motor block effects of ropivacaine in subarachnoid 

space of sheep, also correlating cardiac, respiratory and 

hemogasometric alterations. 

Materials and methods: Six young healthy sheep weighing 

between 47 ± 20 Kg were used. Parameters assessed were heart rate 

(HR), respiratory rate (RR), systolic arterial pressure (SAP), 

sensitive response (SR) and motor response (MR). Arterial blood for 

hemogasometric analysis was also collected. The sensitive response 

was assessed at time zero and after the end of the procedure using a 

needle. During the procedure, surgical stimuli was used to test the 

sensibility. The HR was assessed using cardiac monitor and the SAP 

was measured with a non-invasive method. All the parameters were 

assessed before the administration of the drug (T0) and at 15, 30, 45, 

60, 90 and 120 minutes after. The spinal needle was introducted on 

subarachnoid space between L7 and S1 vertebras. The ropivacaine in 

the dose of 1,5 mg/Kg and volume completed to 10 mL using 

distillated water was administred slowly. The head was maintained 

higher than the body. About 15 minutes later, the animal was 

positioned on its right lateral recumbency and the flank laparotomy 

started. The parameters were assessed until 120 minutes post 

injection and after it, the animal was moved to the recovery room 

and observed until it could stand up. 

Results: There was no statically significant variation of the parameters 

pH, pCO 2, HCO 3, SAP and HR. The RR showed difference at times 30 

to 60 minutes decreasing from the baseline rate. The blockade was 

effective from the tail to the scapular region. 


Conclusions: The experiment showed the hemodynamic stability of 

ropivacaine when used through subarachnoid route. The RR 

diminution may be attributed to an increased RR at time zero, due to 

manipulation stress and direct depression of the respiratory center. 

The sensitive response was annulled from 15 to 180 minutes after 

administration. The motor response, in other hand, had a lesslonging 

blockade, being present again after 90 minutes. This is a 

very interesting characteristic of the drug because it promotes 

immediate post operative analgesia, without the inconvenience of 

the animal being recumbent. Ropivacaine showed many advantages 

over other local anaesthetics when used through subarachnoid route 

in sheep. 

Support: FINATEC 

Key words: anaesthesy, sensitive blockade, motor blockade, flank 

laparotomy, ruminant 

977 Clinical and Radiographic Finding of Lumbar Fracture in a 

Four–Year Old Cow 

A. Ghadrdan-Mashhadi, A. Ghadiri 



Vertebral fractures are common causes of spinal cord injury in large 

animals. Fractures of vertebral column in ruminants may result from 

trauma or abnormal bone mineralization. Spontaneous fracture of 

vertebrae, that are weakened by developmental defects or spinal 

abscesses also common problems in calves. Fractures of the 

lumbosacral spine of cattle are commonly caused by slipping in 

cemented areas. Many of the spinal fracture occur during mounting 

by animals that are exhibiting estral behavior. The clinical signs of 

spinal fractures are variable and depend on the site of the traumatic 

lesion, the severity of the spinal cord compression, and the 

involvement of specific anatomic tracts. A locally bred non-pregnant 

three years old cow was admitted to the veterinary teaching hospital 

of Shahid Chamran (Ahvaz) university with a history of difficulty in 

rising, progressive weakness of the pelvic limbs and ataxia. The 

duration of the disease was two months. The animal kept on grass, 

straw and bran. In clinical examination, the cow was depressed and 

its rectal temperature, heart rate and respiratory rate were 39.3 °C, 

84/min. and 64/min, respectively. On inspection, the lumbar region 

was swollen and on palpation was painful. Left lateral radiography 

was performed. Grid was not used and the exposure factors were 

70KVP and 25mAs. There was a shortening of fourth lumbar 

vertebra with a chip fracture in the ventral part of the body of it. 

According to clinical and radiographic finding, spinal cord 

compression was diagnosed and because the treatment of this disease 

in farm animal is usually not possible, slaughter recommended. 

978 Calf Umbilical Disease: Ultrasound Approach. Field 

Experience 

G. Gnemmi, C. Maraboli 

BOVINEVET, Italy 

Aims: The umbilical calf pathologies are a frustrate diagnosis for the 

veterinary clinician: the size of the calf and the abdomen tension, 

often don’t permit an easy palpation of the umbilicus area, 

consequently the diagnosis of each umbilical tumefaction may be 

mistake, also by an experienced clinician. Aim of this trial was to 

analyze the possible use of an ultrasound unit, normally used in 

bovine reproduction, for the diagnosis of calf umbilical diseases, in 

field condition. 

Material and Methods: The trial has been done during the period Jan. 

2000-Dec. 2007 and it included 206 cases, in 35 farms. We had: 80 

urachal abscess, 12 pervious uracho, 66 hernias (28 complicated), 48 

umbelical vein abscess. The ultrasound examination was performed 

using a 5-7.5 MHz linear probe (Agroscan ® ECM France) and a 5 MHz 

sector probe (L 14 ® ECM France).The area from the sternum to the 

udder was clipped from the hair and a good quality ultrasound gel was 

applied to the skin area.The calves were scanned in standing position,in 

dorsal and in left lateral recumbency. 

Results: Umbilical hernia: has been possible to see if a complicated 

hernia was incarcerated with abomasal or intestinal handle. Urachal 

abscess: has been possible to see the size of the abscess: the abscess 

will appear as granular echoic material inside a dark space (fluid), 

surrounded by a echoic wall, positioned from the apex of the bladder

to the umbelicus. The arteries normally should atrophy after one 

week of age, but in some cases (8 cases) of artery abscessation we 

found two echoic tubular structures caudal to the apex of the bladder. 

Pervious uracho: a fluid-filled sac (anechoic structure with a thin 

wall) with a cord running from the apex of the bladder to the 

umbilicus. Umbelical vein abscess: the abscess is positioned 

between the umbilicus and the liver; the ultrasound exam is 

fundamental in these cases for to determinate the size of the abscess 

and the liver damage, in particularly the ultrasound exam help the 

surgeon to decide if a marsupialization in necessary. 

Conclusion: 

Key words: umbilical, disease, field, ultrasound, approach 

979 General Anesthetic (Propionylpromazine and Xilazine 

Hydrochloride) Usage in Penis Deviation Surgery for 

Marker’s Bulls 

F. Hidalgo y Teran 

Facultad de Medicina Veterinaria y Zootecnia – Unam, Dept. de 

Produccion: Rumiantes, Mexico 

When field testing Bayva1450, 40 years back, the manufacturer told 

us this product was powerful enough to put bovine to sleep. Clearly 

we needed to learn how to use it in bovine practice. At that time we 

used Propionylpromazine as preanaesthetic and sedative, together 

with high amounts of local anaesthetic. They told us could be used 

for minor surgery, and could do without Propionylpromazine 

altogether. However, when we used, we found it had no sedative 

effect; it only provoked chemical arresting and muscular relaxation. 

After recovery, animals were irritated & aggressive. Effect duration 

began at 20 min. and for no more than 45 min. by IM application. 

Propionylpromazine effects began 60 min. after IM application, 

being long lasting with good sedative effect for 7 to 20 hrs. Dosage 

employed was as follows: Propionylpromazine: intramuscular shot, 

2 ml per 100 kg of body weight Xilazine: intramuscular, 1.5 ml per 

each 100 kg body weight In order to assure sought results, maximum 

dosage had to be used, for both drugs. Usually, in the field we were 

able to rig a makeshift operation table out of two iron containers and 

a door. The animal’s head was left hanging free because excessive 

ptialism, and also because ordinarily some ruminal content comes 

out of its mouth. The area around the penis is shaven, as well as 

another previously selected spot. Asepsis is achieved by means of 

10% iodine solution in these parts, and for the surgeon and assistant 

hands as well. Zinc sulfate ointment was used for after-surgery 

protection. An atropine sulfide shot is administered after surgery is 

finished; water is spilt over the animal’s head, which greatly 

contributes to its prompt recovery. Soon the animal wake up and is 

able to walk out of the place by its own means. On these grounds, I 

have concluded that using Propionylpromazine as preanaesthetic, 

and Xilazine as general anaesthetic, excellent anaesthetic effects 

were obtained, effective for 2 hours beginning after 20 min after 

injection of Xilazine; this lapse of time is more than enough for penis 

surgery with having to resort to local anaesthetics, with excellent 

overall results. 

980 Control of Vaginal and Rectal Prolapses by 

Pneumoperitoneum in Sheep - Cases report 

M. Ferreira 1 , V. Gheller 2 , L. Oliveira 2 , I. Borges 1 , G. Macedo Junior 1 

1 Escola de Veterinária da UFMG, Departamento de Zootecnia, Belo 


2 Escola de Veterinária da UFMG, Departamento de Clinica e 

Cirurgia Veterinaria, , Belo Horizonte, Brazil 

Vaginal prolapse is the eversion of the internal structures of the 

female reproductive system as an intussusception-like folding of the 

vaginal floor just cranial to the vestibulovaginal junction. The 

discomfort caused by eversion, associated with irritation and 

swelling of the exposed mucous, results in the increase of the extent 

of prolapse. The consequences can be fatal, depending on the extent 

of involvement of the urinary tract, with narrowing of the urethra and 

bladder rupture. Vaginal prolapse commonly occurs in cattle and 

sheep, usually in adult females during the last months of pregnancy. 

The predisposing factors include increased intra-abdominal pressure 

associated with an enlarged uterus, intra-abdominal fat, increased 

concentrations of circulating estrogen and relaxin during the final 

stage of pregnancy causing relaxation of the pelvic structures and 

perineum, hypocalcemia, high prolificacy, and obesity. Rectal 

prolapse is a common condition that affects sheep, pigs, and cattle. 

The prolapse always results from an increase in intrapelvic pressure 

and therefore accompanies conditions such as tenesmus, diarrhea, 

frequent coughing, obesity, vaginal prolapse, and vaginal irritation. 

Particularly in sheep, excessively short tail docking predisposes to 

rectal prolapse. Amputation of the tail at the sacrococcygeal junction 

results in marked atrophy of the muscles of the tail, which permits 

excessive movement of the rectum to occur. For the reduction of 

prolapse, both vaginal, or rectal, usually proper cleaning and 

reduction of local swelling are done, followed by the reversal of the 

structure and containment of the muscles by Bühner suture around 

the perineum to ensure that another episode does not occur until the 

predicted date of delivery. An alternative technique to control the 

tenesmus and decrease the incidence of recurrence is the permission 

of air entrance within the peritoneal cavity. The technique of 

pneumoperitoneum was used to replace the surgical procedure in 

two confined crossbred LacaunexSanta Inns sheep, which showed 

vaginal and rectal prolapses in the last month of pregnancy. After the 

reduction of prolapses, a 40gauge needle was introduced in the right 

flank of the animals, 2cm below the transverse process and caudal to 

the last rib, allowing the entry of air into the peritoneal cavity and 

reversal of the internal pressure. The animals carry the gestation to 

term, both twin deliveries, without recurrences. 

981 Atlas of Bovine Soft Tissue Endoscopy. New Learning Tool 

for Veterinary Students and Bovine Practitioners 

I. Schlup 1 , M. Wegmüller 2 , M. Stoffel 3 , A. Steiner 1 

1 University of Berne, Clinic for Ruminants, Bern, Switzerland 

2 University of Berne, Virology, Bern, Switzerland 

3 University of Berne, Anatomy and Embryology, Bern, Switzerland 

Introduction: Over the past years, endoscopic examinations in 

veterinary medicine became increasingly important and markedly 

improved the range of clinical diagnostic procedures. Endoscopic 

techniques such as theloscopy, tracheoscopy, rhinoscopy, 

oesophagoscopy, cystoscopy and laparoscopy are of special interest in 

bovine medicine. 

Objectives: In order to make our vast experience in soft tissue 

endoscopy of cattle available to a wider audience, we decided to work 

up our case collection into an electronic atlas. 

Results: At the Clinic for Ruminants of the Vetsuisse-Faculty of 

Berne, Switzerland, an average of 15 endoscopies are performed per 

month, 90% of which are of diagnostic significance. Our 

documentation is based on clinical cases of a three years period (2004- 

2007) and contains more than 60 sequences of pathological findings 

and about 30 sequences (250 screen shots) of physiological situations 

for comparison and some representative examples of endoscopically 

guided surgical interventions. Frequently diagnosed pathologies 

include teat obstructions, retropharyngeal processes and affections of 

the upper respiratory and the urinary tracts. Endoscopically guided 

surgical interventions include theloscopic removal of distal teat 

obstructions and fixation of the abomasum. The electronic atlas is 

based on the popular flash-format, which is suitable for web 

distribution as well as for distribution on DVD. For development we 

used Adobe Flex 2.0, image and video editing was done with Adobe 

Fireworks 8.0 and Adobe Premiere Elements 3.0, respectively. All 

endoscopic examinations were performed with instruments produced 

by Karl Storz Endoskope, Germany (http://www.karlstorz.com). Swiss 

federal animal protection law was strictly respected at all times. 

Conclusions: The electronic atlas represents a suitable learning tool of 

bovine soft tissue endoscopy and endoscopically guided surgical 

interventions for veterinary students and practising veterinarians, 

allowing visual insight in a variety of disease processes. The full 

version will be available on DVD by the end of 2008. A Demo version 

is currently accessible at http://www.wiederkaeuerklinik.unibe.ch/ 

lenya/wiederkaeuerklinik/live/lehre/elektronischelehrmittel.html 

Key words: bovine endoscopy, electronic atlas, theloscopy, 

laparoscopy, oesophagoscopy 

982 Orthopedic Treatment in Ruminants 

E. Tellez, I. Castro 


Veterinaria y Zootecnia, 2nd Department of Large Animals Clinic, 

Mexico City, Mexico 


In Mexico, there is a wide variety of breeds of bovines, goats, sheep 

and wild deer, that are either indigenous to the area or imported from 

other countries. They are used for the production of meat, milk, for 

hunting ranches , in zoos and circuses. Due to injuries and trauma in 

the fields, during transportation, fighting and other causes, these 

animals suffer various types of fractures of long bones such as 

femur, fibula, radius, and also of the metatarsi and metacarpi .The 

Faculty of Veterinary Medicine of the National University of 

Mexico recieves cases from submitted by veterinary practitioners 

from all over the country and also directly from zoos or from animal 

owners everywhere in the country. In the last few years we have 

been asked to reduce fractures using implants such as plates, screws, 

circlage, intramedular pins. In this poster, we present 12 cases 

of traumatized animals, 11 of which were treated with implants and 

one had to be amputated due to the severity of the injury. The group 

included five deer, a wild goat, two bovines (bos taurus) and one bos 

indicus,and three hampshire sheep. The procedures were successful 

in all the deer and sheep and also in one bos taurus, while in the bos 

indicus and one bos taurus of approximately 800 kg and the wild 

goat, the procedure was unsuccessful. The male bulls twisted the 

plates as soon as they stood on their legs and the wild goat died of 

causes unrelated to the surgery. Two of the sheep and the female bos 

taurus were implanted with plates and screws and had a good 

evolution. In the same way, the deer whose fractures were treated 

with Steinman pins had a good recovery. The failures in the bulls 

were due to the excess weight. The Steinman pin worked well in 

animals whose weight ranged between 35 and 40 kg and the plates 

were satisfactory in two sheep that weighed 35 kg and a female bos 

taurus that weighed 225 kg. The experience obtained with these 

cases makes us conclude that plates should not be used in animals 

weighing over 250 kg and that the Steinman pins are very 

satisfactory for use in small domestic or wild ruminants f. e. the 

white tailed deer, goats, sheep and dama fallow deer. 

983 Comparative Efficacy of Flunixine Meglumine in a Tissue 

Cage Model of Inflammation 

O. Roy 1 , P. Cloet 2 , E. Bénizeau 1 , M. Catala 1 , A. Weingarten 3 , 

C. Maisonneuve 3 , D. Sweeney 3 


2 Schering Plough Vétérinaire, Levallois Perret, France 

3 Schering Plough Animal Health, Summit, United States 

The efficacy of Finadyne ® injectable was evaluated for treatment of 

induced inflammation of the subcutis in cattle. It was assessed by 

comparing in a cross-over designed trial the effect of Finadyne ® on 

exudate PGE 2 concentration to that of Metacam ® and 0.9% NaCl 

solution. Twelve healthy calves (183.3 ± 39.1 kg) were included in 

the study and randomly assigned to three treatment sequences. 

Sterile hollow perforated polyethylene balls were surgically 

embedded in the subcutis at four distinct sites per animal two weeks 

prior to D0. On D0, D21, and D42, an aseptic inflammation of the 

subcutis was induced by injecting in three balls per animal 0.5 ml of 

a 2% carrageenan solution. Treatment immediately followed. 0.5 ml 

of exudate was collected prior to challenge, and 2, 4, 8, 12, 24, 36, 

and 48 hours after. Cages were emptied after each collection. 

Exudate PGE 2 concentrations were measured using a validated 

ELISA kit. Each animal received all three items as treatment 

sequentially on D0, D21, and D42. PGE 2 pre-challenge 

concentrations were near 0. PGE 2 concentration levels over time 

displayed a post-challenge sharp increase, peaked 2 hours after 

treatment and gradually decrease, returning to baseline values within 

48 hours. Curves were similar over all three periods.PGE 2 

concentrations when treated with Finadyne ® or Metacam ® were 

consistently lower than those measured when treated with NaCl. 

Percent of inhibition was highest at +4 hours after treatment. 

Differences (absolute values) with the placebo were significant 

throughout the post-treatment period (up to 48 hours) for Finadyne ® 

and at +2, 4, 8, and 12 hours after treatment with Metacam ® . 

Differences in pre-dose concentrations among calves were 

accounted for by dividing post-dose PGE 2 concentrations by predose 

PGE 2 concentrations for each calf. Mean PGE 2 concentrations 

in the NaCl group were significantly higher compared to those in the 

Metacam ® group at 2 and 4 hour, and close to significance at 8, 12, 

24 and 36 hours. They were significantly higher than those in the 

Finadyne ® group throughout the study. At 36 and 48 hours after 

dosing, the mean PGE 2 concentration in the Metacam ® group was 


significantly higher than in the Finadyne ® group. This study shows 

that both items inhibit cyclo-oxygenase with a more potent and 

residual inhibitory activity of Finadyne ® compared to Metacam ® . 

984 A Retrospective Study on 157 Cases of Left Displacement of 

Abomasum (LDA) in Some Dairy Farms in IRAN 

F. Adibhashemi 1 , S. Bokaie 2 

1 Faculty of Veterinary Medicine, University of Tehran, Clinical 

Sciences Department, Tehran, Iran 

2 Faculty of Veterinary Medicine, University of Tehran, Food hygiene 

and Quality Control Department, Tehran, Iran 

Displacement of the abomasum is probably the most common 

surgical condition of bovine patient. Left displacement of abomasum 

is a multi factorial syndrome by many caves. the results and finding 

of studies (cause, number of parturition) may be different in different 

geographic location.The study were done on 157Dairy cattle with 

LDA in some dairy farms in Tehran .The goal of this study is to find 

the answer for some questions (age, sex, number of parturition ) .The 

results showed that 96.5% of cases were in female and 3.2% in male 

sex. The other results were as follows: 2.6% of cases were heifers 

25% in first parturition, 7.2% in 2th parturition 35.5% in 3th 

parturition, 25% in 4 th parturition 4.6% in 5 th parturition 1.3% of 

cases were in before parturition 0.7% in first week after parturition 

2.07% in 2 weeks after parturition 50% in 3 weeks after parturition 

10.5% in 4 th weeks after parturition 15.8% in 5 th weeks after 

parturition 8.06% of cases recovered after surgery completely 5.3% 

were culled for some complications 13.7% of cases were culled for 

accompany diseases . 

Key words: left displacement of abomasum, cattle 

985 Comparison of Intranasal Versus Intravenous Lidocaine 

Application during Rhinotracheobronchoscopy in Cattle 

A. Dadak 1 , S. Franz 2 

1 Institute for Pharmacology and Toxicology, Vienna, Austria 

2 Clinic for Ruminants, Vienna, Austria 

Objectives: Research was done on the efficacy and safety of lidocaine 

during endoscopic procedures of the upper respiratory tract in cattle. 

Patients and Method: Experiments were performed in healthy non 

lactating cows. Eight minutes after intranasal (IN) or intravenous (IV) 

application of lidocaine (1.5 mg/kg), both given over a period of 5 min, 

rhinotracheobronchoscopy was performed. Experiments were 

videotaped for subsequent analysis of behavioural changes. Mean 

invasive arterial blood pressure (MABP) and heart rate (HR) were 

analysed at specified time points. Blood samples were taken up to 60 

min in order to measure serum lidocaine concentrations. 

Results: Preliminary data show that lidocaine did not lead to 

significant cardiovascular reactions in cattle, neither after IN nor 

after IV administration. In the present study absorption of lidocaine 

was generally rapid after IN application. During the procedure of 

rhinotracheobronchoscopy, HR and MABP increased slightly in all 

groups (lidocaine IN, IV and the control groups). Lidocaine 

significantly improved the animals’ tolerance during rhinoscopy, 

especially when given IN. Insertion of the endoscope into the trachea 

was eased in both, the lidocaine IN and the lidocaine IV group. 

Discussion and Conclusion: We conclude from our results that 

lidocaine is a safe and effective analgesic option for endoscopic 

procedures of the upper respiratory tract when given at a dosage of 

1.5 mg/kg. On the basis of our results we favour intranasal over 

intravenous application of lidocaine for rhinotracheobronchoscopy 

in cattle. 

Key words: cattle, endoscopy, lidocaine, application 

986 Study of Toxicological Effects of Epidural Neostigmine 

Methyl Sulfate in the Cow 

S. Zolhavarieh 1 , S. Zolhavarieh 2 

1 Junior School of Veterinary Medicine, Bu-Ali Sina University, 

Clinical Sciences, Hamedan, Iran 

2 Hamadan University of Medical Sciences, Clinical Sciences, 

Hamedan, Iran 

Background: The spinal delivery of the cholinesterase inhibitor 

neostigmine yields analgesia in rats and augments the analgesic

effects of alpha 2 agonists in sheep. To assess its activity in cow, 

preclinical toxicology studies to define its safety were required in 

this species. 

Methods: For this study, female cows were prepared following rigid 

aseptic precautions epidural injections. Cows epidural received daily 

injections of 0.025 ± 0.040 mg/kg neostigmine (n = 6/group) for 4 days 

and were observed for general behavior and analgesic effect via pick of 

perineal region with a needle and second group received normal saline 

at the same volume of neostigmine. 

Results: In cows, neostigmine produced a dose-dependent increase 

analgesia in needle test, and no tolerance was observed. Mild tremor 

was observed but was not debilitating. A mild inflammatory reaction to 

the needle in the site of injection was observed in both groups. In cows, 

neostigmine produced bradycardia and an increase in muscle tone. 

Cerebrospinal fluid protein, specific gravity, and glucose were elevated 

in neostigmine group. No group differences in general condition were 

observed. 

Conclusion: These results suggest that, there is no evidence of toxicity 

that can be attributed to the neostigmine. 

Key words: epidural, neostigmine, toxicology, cow 

987 Preanesthetic Butorphanol Improving Analgesia during 

Laparoscopy for Follicular Aspiration in Goats 

M. Cordeiro, W. Vicente, D. Dias, M. Oliveira, P. Di Filippo 

Universidade Estadual Paulista Júlio de Mesquita Filho, Animal 

Reproduction, Jaboticabal-SP, Brazil 

The aim of this study was to evaluate the influence of butorphanol in 

she-goats anesthetized with total intravenous anesthesia undergoing 

laparoscopy for follicular aspiration. Twenty female adult goats 

(aged 12-36 months, weighing 24 ± 6 kg) received a combination of 

xylazine (0,05mg/kg, IM) and butorphanol (0,5mg/Kg, IM) like 

preanesthetic medication. General anesthesia was induced with 

ketamine (2 mg/kg, IV) and maintained with a Total Intravenous 

Anesthesia (TIVA) composed by guaifenesin (50mg/mL), xylazine 

(0,1mg/mL) and ketamine (1mg/mL) diluted in 5% dextrose/0,9% 

saline solution (GXK). Endotracheal intubation was performed and 

animals were connected to circular breathing circuit during 

spontaneous ventilation (O2 flow rate: 10mL/kg/min). The GXK 

infusion rate started at 2mL/kg/h and was later adjusted according to 

clinical evaluation of anaesthetic depth. Heart rate (HR), respiratory 

rate (RR) and analgesia were evaluated at the following moments: 

T0 (immediately after induction); T1 (immediately after beginning 

of TIVA’s infusion); T2 (immediately after abdominal insufflation 

with CO2); T3 (15 min after T2); T4 (15 min after abdominal 

decompression and drugs infusion end). All animals showed 

analgesia and immobilization allowing the accomplishment of the 

procedure. It’s impossible to determine an exact TIVAs infusion rate 

because of individual parameters variations during the surgery. We 

could observe a RR increase at T2 and T3, when compared with T1. 

The RR increase was following by a decrease in the respiratory 

amplitude, however HR had not changed. The RR increase was 

induced by diaphragmatic compression due to Trendelenburg 

position and due to abdominal CO2 distention, both fundamental 

procedures to surgical management. The results suggest that the use 

of butorphanol in pre-anaesthetic medication was efficient to 

improve the analgesia during TIVA’s infusion allowing the 

follicular aspiration by laparoscopy in this specie. We concluded that 

this anaesthetic protocol is practical, effective, safe and can 

undoubtedly be recommended for surgical procedures that requires 

general anaesthesia in goats. 

Key words: TIVA, butorphanol, laparoscopy, goat, pre-anaesthetic 

medication 

988 Abomasal Impaction due to Sand Ingestion in a Dairy Cow 

J. Peiró 1 , F. Feitosa 1 , L. Mendes 1 , F. Cadioli 1 , G. Soares 1 , 

M. Araújo 2 , P. Teodoro 2 

1 Sao Paulo State University - Aracatuba Campus, DCCRA, 

Aracatuba, Brazil 

2 Veterinary Teaching Hospital, Aracatuba SP, Brazil 

Objectives: To report a case of an abomasal sandy impaction in a dairy 

cow. 

Case description: One severe case of sand impaction occurred in a 

small dairy herd. Diet was fed as concentrate exclusively (sorghum 

silage) on the ground. There was no pasture available in the property 

due to the dry season. A Holstein cow, 5 years old, had decreased 

food intake, low body score, ´papple-shaped´ abdomen, signs of 

rumenal tympany, and dark feces. The cow was referred to the 

Veterinary Teaching Hospital. The orogastric tube was passed to 

eliminate ruminal gases, unsuccessfully, and the ruminal pH was 8. 

Physical exam revealed increased heart and respiratory rates, and 

low rectal temperature. Congested mucous membranes, dehydration, 

and incomplete ruminal movements in 5 minutes were observed. The 

metal detector did not detect the presence of metallic materials. 

Right flank laparotomy was performed, and jejunal volvulus and 

sand impaction of the rumen and abomasum were diagnosed. After 

volvulus correction, the animal received antibiotic treatment. The 

cow showed abdominal distension 4 hours after surgery and a rumen 

fistula, using a plastic syringe, was produced to eliminate the gas. 

Five days after surgery, the animal showed abdominal pain, bilateral 

abdominal wall distension, dark feces, remained in sternal 

recumbency, and died 24h later. 

Conclusions: Due to the carbohydrate-rich and fiber-poor diet, the 

animal developed ruminitis and food fermentation in different parts 

of the gastrointestinal tract. This maybe was responsible for the 

small intestinal volvulus. The clinical picture deteriorated fast due to 

the reluctance of the owner to refer the animal to the Veterinary 

Hospital because he believed it was a simple case of ruminal 

tympany. 

Key words: sand impaction; bovine 

989 Quantifying Some Causes of Visceral Pain in Cattle 

P. Scott 

University of Edinburgh, 2- Department of Large Animals Clinic, 


Evidence of pain is an important clinical feature of many 

inflammatory conditions involving various thoracic and abdominal 

organs in cattle particularly when the pleurae and peritoneum are 

involved but the extent to which expression, pulse, respiratory rate 

and effort, rumen motility, stance, gait, and production parameters 

reflect the severity of the condition remains largely 

unquantified. Diagnosis by further increasing pain (withers pinch 

test, ballotment) is still considered the most reliable indicator for 

traumatic reticulitis. Ultrasonography has greatly increased the 

practitioner’s ability to confirm early peritoneal reaction, pleurisy 

and pericarditis. A quick and simplified protocol for 

confirming/quantifying these diseases is required, thereafter 

treatment strategies and provision of analgesia can be objectively 

evaluated. Methodology Diagnostic quality images are readily 

obtained using a 5.0 MHz sector transducer connected to a real-time, 

B-mode ultrasound machine but a linear scanner will suffice in most 

situations. 

Results: Peritonitis of the anterior abdomen involving the reticulum, 

the classical wire scenario, is readily identified with initial 

accumulation of peritoneal exudate, followed by fibrinous adhesions, 

and abscess formation in some cases. Marked intestinal distension is 

a feature of focal peritonitis involving the small intestine. Diagnostic 

procedures such as abdominocentesis can be useful but do not 

indicate the extent of inflammation/infection.Pericardial and pleural 

effusion/infection are readily distinguished during ultrasonographic 

examination. Fibrinous pleurisy appears as a hypoechoic layer 

separating the pleurae and has been diagnosed in 

pasteurellosis/shipping fever. Necrotising pneumonia, inhalation 

pneumonia and pleural abscess show obvious pain but cannot easily 

be differentiated from endocarditis and anterior abdominal infections 

on clinical examination alone. Cattle with chronic suppurative 

pulmonary disease show less obvious pain but can be diagnosed by 

demonstrating marked disruption of the visceral pleura with 

hypoechoic areas (often consolidation), extending up to 8 cms often 

with a lobular distribution. 

Conclusion: A single injection of a soluble corticosteroid, such as 

dexamethasone, is preferred to NSAID for inflammatory pain. 

990 Intracardial Pressure in Relation to the Venous Pulse and 

Venous Tension in Healthy and Heart Compromised Cows 

S. Eisenberg, A. De Vos, A. Grootendorst, L. Van Leengoed 

Faculty of Veterinary Medicine, Farm Animal Health, Utrecht, 



Cardiovascular disorders are commonly found in dairy cattle. Clinical 

examination of the circulatory system to qualify heart function will 

reveal increased venous tension and presence of the jugular venous 

pulse. Both, jugular vein and mammary vein are manually examined to 

determine the central venous pressure (CVP). In patients with 

progressive decrease of cardiac capacity CVP is often increased, and 

leads to palpable veins and a jugular venous pulse that reaches the arch 

of the lower jaw and a pulse that can be counted at the mammary vein. 

The aim of this study was to examine the relatedness between results 

from clinical examination and results from invasive measurement of 

intracardial and pulmonal pressures. Eleven cows were both clinically 

examined and their blood pressure was measured invasively by Swan- 

Ganz. Procedures were carried out in 3 clinically healthy cows (1), 2 

adult cows with a clinically diagnosed ventricular septal defect (VSD); 

2), 3 cows with a clinically diagnosed endocarditis (3) and 3 cows with 

a clinically diagnosed pericarditis (4). Diagnosis of group 3 and 4 were 

confirmed by post-mortem examination. Venous tension (jugular vein) 

was increased in all cows of group 3 and in all cows of group 4. A 

pulsing mammary vein was palpable in 1 cow of group 3 and 2 cows of 

group 4. None of the cows of group 1 and of group 2 showed any signs 

of elevated CVP. The intracardial pressure curve of 1 cow of group 3 

showed an unusual form of the jugular pulse, an increase of the c-wave, 

which could be interpreted as a leakage of the RAV valves. The 

intracardial pressure curves of cows of group 3 were unusual. One curve 

showed increased a-waves of the jugular pulse giving the impression of 

a stenosis of the RAV valve, the other two showed an wedge curve with 

increased waves which might be du to leakage of the LAV valves. The 

post-mortem of group 3 revealed cardiomypathy in one case, 

endocarditis of the aortic and pulmonic valves in one cow and 

endocarditis of all valves in the third cow. Post-mortem results of group 

4 confirmed pericarditis and no macroscopic lesions of the heart 

valves. In this study cows with pulsing mammary veins had an elevated 

CVP except one cow with a VSD. A pressure curve of the right atrium 

316 Fungal and Mycotoxins Contamination in Food, Feed and 

Milk Samples from Selected South African Rural Areas: a 

Really Health Issue? 

M. Mulunda, M. Faneli, M. Dutton 

University of Johannesburg, Food, Envirronmental and Health 

Research Group, Johannesburg, South Africa 

Mycotoxins are secondary metabolites of fungi, which may 

contaminate feed and food at all stages of the food chain. Their 

global occurrence is considered as an important risk factor for 

human and animal health, as up to 25% of the world crop production 

may be contaminated with mycotoxins. The main objective of this 

study was to evaluate food and feed quality with respect to fungi and 

their mycotoxins in selected rural areas of South Africa. In this 

study, 100 samples of animal feed, 150 of staple foods (Maize, corn, 

porridge, nuts) and 50 samples of fresh milk obtained from cattle and 

goats were analysed for fungi and mycotoxins contamination. All 

feed and food samples were cultured and sub cultured on Ohio 

Agricultural Experimental Station agar (OAESA) or Potato Dextrose 

Agar (PDA), Czapek 20 (CY20S), Czapek (CZ) and Malt extract 

agar (MEA) under aseptic conditions and incubated at 28 °C for 4 to 

7 days for fungi isolation and identification. The multi mycotoxin 

extraction method, the thin layer chromatography (TLC), VICAM 

immunoaffinity column, high performance liquid chromatography 

(HPLC), gas chromatography mass spectrophotometry and the 

ELIZA immuno assay kits were used for mycotoxin determination 

and quantification and confirmation. Aspergillus niger, A. clavatus, 

A. flavus, A. fumigatus, Fusarium verticillioides, F. graminerium 

and F. proliferatum were the most prevalent contaminant fungi in 

100 % of analysed samples. Fumonisin B 1 and B 2 (0.4 - 800 µg/kg), 

aflatoxin B 1 (0.13-61ppb), zearalenone (0.001-11ppm) and 

deoxynivalenol (0.2-11ppm) were found, respectively, in 68%; 76%; 

78% and 74% of analysed samples. The methods used for mycotoxin 

analysis were found to be reliable as most of the results received by 

various kind of methods were found to be in good agreement. 


Food Safety 

with a higher c-top, probably caused by leakage of the RAV valves, was 

found in only one animal with a venous pulse of the mammary vein. 

Key words: central venous pressure, cardiac failure, cattle 

991 Ear Trimming in a Neonate Lamb due to Pyogranulomatosis 

A. Raayat Jahromi, A. Meimandi Parizi, A. Khodakaram Tafti 

School of Veterinary Medicine, Shiraz University, Shiraz, Iran, 

Clinical Sciences, Shiraz, Iran 

Ear surgeries are uncommon in ruminants, because of drugs are 

effective enough in most ear injuries; and only in rare conditions 

surgery is needed. Ear brief injuries in ruminants whit no 

complications will not have a serious consequent; but infections may 

interfere with auditory abilities. A neonate lamb with a very big and 

thickened ear was referred to Veterinary teaching hospital of School 

of Veterinary Medicine of Shiraz University. A small superficial 

wound had occurred in convex surface of ear two weeks ago and 

only disinfection of the wound with no other treatments was done. 

After some days, depression, anorexia and head shaking were 

observed followed by more enlargements in affected ear. In clinical 

examination, the ear was too swelled so that the head was deviated to 

the same side. Its external surface was keratinized and the lesion was 

extended to near the external ear canal. In clinical examination; heart 

rate and respiratory rate were normal but rectal temperature revealed 

a moderate fever. Under general anaesthesia, the ear was fully 

trimmed and the edges were sutured by simple interrupted of 0 nylon 

and the external ear canal left open. In microbiologic and pathologic 

studies, Staphyloccus aureus (coagulase positive) was isolated from 

the mass and pyogranulomatous dermatitis was reported. Penicillin 

and flunexin meglumin were administered for 3 days postoperative. 

There was no complications in 2 months postoperative follow up. 

Key words: ear trimming, lamb, pyogranulomatosis 

Aflatoxin M 1 was detected at 25.0-108.4 ng/l in 100% of milk 

samples on ELIZA and 80% on VICAM test. Traces of fumonisin B 1 

(0.1-0.2ng) were detected in 2 % of milk samples tested. Finally a 

cytotoxicity essays were carried on lymphocytes using fumonisins 

and aflatoxins standards and extracts with positive dose response 

toxicity results. The results obtained from this study might be among 

factors contributing to health problems faced by animals and human 

population due to chronic exposure to these contaminated feed or 

food characterised by immunosupression, abortion, infertility, 

mycosis, liver and oesophageal cancer inboth animals and human. 

Key words: mycotoxin, fungi, cytotoxicity, health 

317 Virulence-, and PFGE Profiles of Verotoxigenic and 

Enteropathogenic Escherichia coli O157 Strains in Hungarian 

Cattle 

I. Tóth 1 , G. Kardos 2 , H. Schmidt 3 , J. Pászti 4 , B. Nagy 1 

1 

Veterinary Medical Research Institute of the Hungarian Academy of 

Sciences, Enteric Bacteriology and Foodborn Zoonoses, Budapest, 

Hungary 

2 

National Veterinary Institute, Microbiology, Debrecen, Hungary 

3 

University of Hohenheim, Food Microbiology, Stuttgart, Germany 

4 

National Center of Epidemiology, Phage and Molecular Typing, 


Vetoroxin producing Escherichia coli (VTEC), of ruminant origin 

have been implicated in epidemic or sporadic cases of 

enterohaemorrhagic diseases (EHEC), and haemolytic uremic 

syndrome (HUS) of man in several countries. In Hungary, only 

sporadic cases have been identified from mainly unknown sources 

due to the predominant E. coli serotype O157:H7. In this study we 

wanted to reveal the nature and frequency of bovine sources of 

possible human infections by a cross sectional microbiological study 

of one large dairy herd and of 4 abbatoirs in Hungary. For this 

purpose faecal and raw milk samples of the large dairy farm and 

rectal and colon samples of cattle from the abattoirs were tested,

using immuno-magnetic and pre-selective methods on samples from 

a total of 542 cattle. 

Results: 41 E. coli O157 (plus, 13 non-O157 VTEC) strains were 

isolated (1% of cattle). 

The O157 strains formed the following 3 pathogroups: 

- EPEC (enteropathogenic E. coli) O157:H7 (22 strains) (eae + , stx - ),- 

EHEC (enterohaemorrhagic E. coli) O157:H7 (11 strains), (eae + , stx + ), 

- atypical O157 (8 strains), (eae - , stx - ), characterized by genes of the 

cytotoxin CDT-V.The E. coli O157 strains frequently harboured long 

polar fimbrial (lpf O113 and lpf R141) genes and pathogenicity island OI- 

122 genes (including efa, sen and pagC). The fyuA, marker gene for 

high pathogenicity island (HPI) was detected in some strains. Both 

O157:H7 EPEC and EHEC possessed - type of intimin (eae) gene and 

- type of translocated intimin receptor (tir) gene (characteristic for 

human O157:H7 EHEC strains). 

Pulsed field gel-electrophoresis (PFGE) revealed three clusters of 

bovine O157:H7 strains: 

- O157:H7 EPEC strains from large intestine of slaughtered cattle 

(Cluster I), 

- O157:H7 EHEC strains of the above slaughtered cattle (Cluster II), 

- O157 atypical strains from raw milk samples of the dairy farm 

(Cluster III) 

This grouping was partly confirmed by multi locus sequence typing 

(MLST) performed on five housekeeping genes. 

Conclusions: it seems that O157:H7 strains are not infrequent in the 

Hungarian cattle population. They represent EPEC and EHEC, 

possessing virulence genes with zoonotic potential, although human 

infection due to bovine EPEC has not been described yet. Our data 

indicate that beside EHEC, the bovine O157:H7 EPEC and atypical 

O157 E. coli strains deserve more attention in the future. 

318 Comparison of Conventional Method and Microspheresbased 

Assays for the Quantification of Kanamycin in Milk 

L. Chen 1 , Y. Jin 2 , M. Zou 2 , X. Cui 1 , X. Hu 1 , B. Han 1 

1 

China agricultural University, College of Veterinary Medicine, 

Beijing, China 

2 

Chinese Academy of Inspection and Quarantine, Institute of 

Inspection, Beijing, China 

Several techniques had been developed to quantify antibiotics in cow 

milk; however, all of them possessed one or more significant 

limitations and usually measured only one analyte at a time. 

Competitive ELISA, which showed great correlation coefficient over 

the range from 1ng/ml to 100ng/ml, was the most conventional assay 

for quantifying low-molecular-weight compounds like antibiotic. 

Immunochromatographic strip test, which could detect samples with 

naked-eye, was more convenient but had a high detected limitation. 

HPLC (high performance liquid chromatography) was more 

reproducible, whose precision of within-day assay showed a 0.5 to 

4.0% coefficient of variation (n=5) when the kanamycin concentration 

ranged from 1x10 4 

to 7x10 4 ng/ml. Although CE (capillary 

electrophoresis) analysis could guarantee a good linearity in the 

concentration range 400-5000 ng/ml with a detection limit of 

100ng/ml, its intra-day and inter-day RSD (relative standard deviation) 

were 3.4 and 11.2%, respectively, and its recoverability was as low as 

approximately 60%. Currently, we used a novel multiplexed method 

improved on the existing assay to measure kanamycin whose 

concentration was from 0.1ng/ml to 3 ng/ml, and its intra-day and interday 

RSD could reach to 3.4 and 4.1%, respectively. In this method, 

antigens were covalently bound to carboxyl-modified microspheres, 

which could be distinguished by their distinct proportions of red and 

orange fluorescent dyes, and then were captured by monoclonal 

antibody against kanamycin in milk samples. The main advantages of 

such a method lay in its high efficiency, good biocompatibility, 

excellent selectivity as well as ultra-low sample consumption, and it 

showed a great potential for multiplex assay in the near future. 

Key words: kanamycin, flow-cytometry, cow milk, ELISA, CE, 

HPLC, multiplex assay 

319 Quantitative Risk Assessment Model of Human Salmonellosis 

linked to the Consumption of Camembert Cheese made from 

Raw Milk 

A. Fares, A. Dayhum, M. Sanaa, O. Cerf, Y. Millemann 

Ecole Vétérinaire d'Alfort, Unité Epidémiologie et Analyse de Risque, 

Maisons-Alfort, France 

Salmonellae are one of the most important causes of foodborne 

illness associated with raw dairy products. The assessment of the 

real risk associated with the consumption of these products is needed 

and the most appropriate method to achieve this goal is the risk 

analysis process, which links pathogens in food to the public health 

problem. The main aim of this work was the quantitative assessment 

of the risk of human salmonellosis linked to the consumption of 

Camembert cheese made from raw milk cheese. A data gap that is 

routinely identified in risk assessment is the lack of quantitative data 

on pathogens contaminated food; therefore, we developed a rapid, 

sensitive and reliable method for the quantification of Salmonella in 

artificially contaminated milk samples. The method combined the 

principles of most-probable-number (MPN) method with a real-time 

PCR assay. With this developed assay (MPN-real-time PCR) low 

levels of Salmonella (1-5 CFU/mL) in milk could be enumerated 

after 8 h of non-selective enrichment in buffered peptone water. This 

developed quantification assay was applied to naturally 

contaminated bulk tank milk samples collected from dairy farms 

located in western France. Eight (2.6%) of 299 bulk tank milk 

samples were found positive, with estimated MPN values ranging 

from 3.7 to 79.2 MPN/mL of milk. In the assessment of potential 

risks associated with Salmonella in raw milk and raw milk products 

it was necessary to examine the ability of Salmonella to grow in 

milk. Primary and secondary models, describing mathematically the 

growth of two Salmonella strains (S. typhimurium and S. tontevideo) 

in milk under constant temperatures during different incubation 

periods, were developed. The primary logistic-with-delay model was 

used to describe Salmonella strains growth as a function of time. The 

specific growth rates of S. typhimurium and S. montevideo varied 

according to serotype and temperature. Both the primary and 

secondary models fitted growth data well with a high-pseudo R 2 

(0.97-99). Finally, a quantitative risk assessment of human 

salmonellosis linked to the consumption of Camembert cheese made 

from raw milk is presented based on the results of the previously 

mentioned objectives of this thesis. Different distributions were 

assumed for parameters of the model and a Monte Carlo simulation 

was used to model the process and to quantify the risk associated 

with the consumption of 25 g serving of cheese. 

Key words: salmonella, raw milk, risk analysis 

320 The Seasonal Changes of Contamination Rate of Ovine Meats 

to Salmonellae Bacteria Serotypes in Kermanshah 

Dr. Bahiraie, Dr. Chale chale, Dr. Pouanmehr 

Razi University,Faculty of Veterinary Medicine, Kermanshah, Iran 

Objectives of study: The survey of contamination rate of ovine meats 

(meats of sheep) in Kermanshah city. The comparisons of 

contamination rate between two types of meat which delivered to 

customers include absolute and grinded meat. The study and 

comparison of differences of contamination rate during 4seasons of 

year. The salmonellae serotyping in contaminated meats. The survey of 

Serotypes Prevalence in each season. 

Material and Methods: Bacteriological examination was carried out 

on two sample groups of ovine meat (500 absolute meat and 500 

grinded samples) .The samples were collected from meat markets of 

city during four season of year (2005). Each was 50 grams of meats 

which prepared to customers. The samples were were examined in 

regarding to routine bacteriological examinations. Stereotyping by test 

of Fast agglutination on slid and were confirmed by tube tests. Data 

analysied in SPSS .Serotypes of salmonellae based on Kaffman-white 

schema were identified. 

Results: Salmonellae were isolated 31 out of 500 ovine absolute 

meat samples (A.M.S) (%6.2) and 43 out of 500 ovine grinded meat 

samples (G.M.S) (%8.6) totally during the year. The highest 

contamination rate for A.M.S was with 13 cases in summer, 9 

cases in fall, 7 cases in spring and 2 case in winter, and for G.M.S 

samples was 17 cases in summer, 12 cases in fall, 10 cases in spring 

and 4 case in winter, respectively. Stereotyping showed that: in two 

groups of contaminated samples the S.typhimurium was the most 

contaminator serotype The rates of contaminator serotypes for 

absolute samples (A.B.S) were S. typhimurium (28.7 %) and then S. 

enteriditis and for G.M.S were S. typhimurium (34.3 %) and then S. 

enteriditis (22.4 %), respectively. S. paratyphi B, was found in 

G.M.S 2 cases in fall and 1 case in summer. Distribution of the 

serotypes of salmonellae in grinded samples G.M.S was more than 

absolute meat samples. 

Food Safety 269

Conclusion: The total contamination rate of salmonellae is variable 

during seasonal changes. The grinded meats (8.6%) were more 

contaminated as compared with absolute meats (6.2%). The S. 

typhimurium was the most contaminator serotype of salmonellae in 

ovine meatsThe presence of S. paratyphi B serotype in grinded meats 

might be explained by spread of contaminations from human 

vectors. 

Key words: meat hygiene, microbiology, salmonella bacteria 

321 Vaccination of Cattle with E. coli O157:H7 Type III Secretion 

Proteins as a Pre-slaughter Intervention Method to Reduce E. 

coli O157:H7 Prevalence 

D. Rogan 1 , D. Smith 2 , R. Moxley 2 , A. Potter 3 , J. Yome 1 

1 

Bioniche Animal Health Canada, Research & Development, 

Belleville, Ontario, Canada, Canada 

2 

University of Nebraska-Lincoln, Research & Development, Lincoln, 

Nebraska, United States 

3 

Vaccine & Infectious Disease Organization, Director and CEO, 

Saskatoon, SK, Canada 

Enterohaemorrhagic Escherichia coli O157:H7 (EHEC) is an 

important pathogen of humans, causing severe diarrhea and 

hemolytic uremic syndrome (HUS). In North America, EHEC 

serotype O157:H7 is most prevalent, causing 85-95% of HUS cases. 

Cattle are an important resevoir of E. coli O157:H7 and as such, are 

a major source of infection in humans, through direct contact or the 

consumption of contaminated meat, water, or produce. E. coli 

O157:H7 infection of cattle requires type III secreted proteins 

(TTSP) which enable the bacteria to colonize the intestinal mucosa. 

Bioniche Animal Health has developed an E. coli O157:H7 TTSP 

vaccine that contains the TTSP proteins EspA, EspB and Tir. The 

efficacy of this vaccine has been evaluated in cattle using natural 

exposure and controlled challenge studies. To assess the effects of 

vaccination on the shedding of E. coli O157:H7, calves were 

vaccinated with three doses and challenged with E. coli O157:H7 14 

days later. There was a 2.28 log 10 reduction in the magnitude of 

shedding in vaccinated animals compared to controls. In another 

study, with a three dose vaccination and natural exposure to E. coli 

O157:H7, vaccinated animals were less likely to be colonized by the 

bacteria in the mucosa of the terminal rectum at harvest (OR=0.014, 

p

Campylobacter spp. at each sampling point tested was 1.7%, 3.3% 

and 0.0% while the incidence of Salmonella spp. at each sampling 

point was 0.0%, 5% and 1.7%, respectively. The results indicate 

that very few cattle carcasses in Shahrekord appear to carry any of 

the pathogens sought. 

Key words: campylobacter, salmonella, cattle 

1212 Study on Infestation of Cattle Carcasses to Sarcocyst spp in 

Slaughtered Cows in Shahrekord by Impression Method 

M. Boniadian 1 , B. Meshki 2 

1 

Veterinary Faculty of Shahrekord University, Food Hygiene, 


2 

Facutly of Veterinary Medicine, University of Tehran, Department 

of Parasitology, Tehran, Iran 

To determine the macroscopic and microscopic cysts of Sarcocyst 300 

carcasses (138 male & 162 female) were inspected and samples were 

collected from muscles of heart, esophagus, diaphragm, triceps and 

shoulder, Daub smear were prepared and after staining were studied 

with microscope to find the microscopic cyst of parasite. X 2 test was 

used to find the relation between the rate of infestation and age, sex and 

different muscles. 

Results: Gross examinations declare in none of the samples, but 

daub smear method showed microscopic cysts in 91% of the 

samples. There was no significant difference between male and 

female for microscopic cysts (P>0.05). Also in the cases with 

microscopic cyst, no significant difference was observed in different 

ages (P>0.05). The results showed that there is significant difference 

among different muscles with microscopic cysts (P

Material and methods: The trial included 18 Czech-Pied bulls fed 

a diet with a high proportion of concentrated feed and 18 Czech-Pied 

bulls fed a diet based on maize silage, used as a control group. The 

trial was launched after the weaning of calves. During the fattening 

period, live weight and average daily weight gain were monitored. 

The bulls were slaughtered at the live weight 550 - 600 kg, mean age 

at the slaughter was 473 days for the high-grain diet group and 474 

days for the control group. The carcasses were classified to SEUROP 

quality grades, and net weight gain and carcass yield were 

calculated. Samples of m. longissimus pars thoracis were withdrawn 

from five bulls in each group to examine selected meat quality 

parameters. 

Results: In the period from weaning to slaughter, the high-grain diet 

bulls and the control bulls achieved average daily weight gain of 

1.29 kg and 1.21 kg, respectively. Differences between the groups 

were not significant. The high-grain diet group showed a higher 

average carcass weight and higher net weight gain, differences 

between the groups were not significant, too. As to meat quality 

parameters under study, a significant difference was found only in 

meat lightness (L*), with the mean value in the high-grain diet group 

being significantly (P < 0.01) lower than in the control group. Other 

meat quality parameters did not show any significant differences 

between the groups. 

Conclusions: In this study, the high-grain diet gave similar 

performance as the maize silage based diet in fattening bulls. The highgrain 

diet group and control group showed comparable average daily 

weight gain and selected carcass and meat quality parameters. 

Key words: concentrated feed, weight gain, carcass weight, m. 

longissimus 

1217 Quantification of Salmonella in French Cattle Fecal Samples 

at Slaughter House by MPN Real Time PCR 

A. Dayhum, A. Fares, M. Sanaa, O. Cerf, Y. Millemann 

Ecole Vétérinaire d'Alfort, Unité Epidémiologie et Analyse de Risque, 

Maisons-Alfort, France 

As a part of our effort in quantitative risk analysis of food-borne 

diseases, the objective of this study was to develop rapid and reliable 

protocols for detection and quantification of Salmonella in fecal 

samples at slaughter house. First, for the detection of Salmonella in 

artificially and naturally contaminated fecal samples, SYBR Green I 

real-time PCR assay was used, where quantification of Salmonella 

was achieved by combining this assay with most-probable-number 

(MPN) method (MPN-real-time PCR). To develop or test this 

protocol for detecting and enumerating of Salmonella in artificially 

contaminated fecal samples, a Salmonella enterica serotype 

Typhimurium DT104 strain was inoculated into fecal samples at 

different levels of contamination. Data on artificially contaminated 

fecal samples indicated that both detection and quantification 

protocols were able to detect and enumerate as few as 1 CFU/mL of 

fecal after 8-h of a single non-selective pre-enrichment step in 

buffered peptone water. All MPN estimates corresponded well to 

inoculum levels. The protocol was then applied to naturally 

contaminated fecal samples. A total of 296 fecal and 26 

environmental samples were aseptically collected from 

slaughterhouse located in Meaux, France weekly during February to 

March 2006 (an average of 40 samples per visit). Salmonella was 

positive from 9.12% (27/296), 34.62% (9/26) fecal and 

environmental samples respectively, with estimated MPN values or 

counts of Salmonella ranging from 1400 MPN/g), while all the other 21 animals were less than 80 

MPN/g (from them 13 animals 0.05) Also the most important cause 

of microbial contamination and high microbial load of 

hamburgers was belong to contamination of meat and carcasses in 

slaughterhouses due to poor and unhygienic handling 

and undesirable sanitary situation during production and distribution 

of meat. 

Key words: hamburger, microbial quality, Iran 

1219 Comparative Survey on Hygienic Quality (Coliform , 

Escherichia coli and Staphylococcus aureus) of Industrial 

Butters with Using Standard Methods and Impedance - 

Splitting Method 

A. Fazlara 1 , A. Khataminia 2 

1 

Shahid Chamran University, Food Hygiene and Quality Control 

Department, Ahvaz, Iran 

2 

Veterinary Organization, Ministry of Jihad-e-Keshavarzi, Ahvaz, 

Iran 

This study was conducted to compare impedance - splitting method 

and conventional standard (reference) methods to evaluate microbial 

quality of produced pasteurized butters in Ahvaz area in Khuzestan 

province of Iran. In this study, 90 sample of pasteurized butter from 

different batches were collected during warm and cold seasons (from 

October 2006 to September 2007) from market places in Ahvaz city 

and tested for coliforms, Escherichia coli and Staphylococcus aureus 

by both methods. The standard methods were based on references 

and standard institute and industrial investigation of Iran’s 

recommendation and the impedance method was done by measuring 

the variation of electrical resistance of broth media (M-value) and 

electrodes (E-value) for detection of coliform, Escherichia coli and 

Staphylococcus aureus respectively. According to the results, there 

was no contamination to coliforms and Escherichia coli in 

pasteurized butters and the both methods were totally correlated and 

the specifity of impedance method was 100 percent. Also 5 (5.5%) 

and 3 (3.3%) of all samples were contaminated to Staphylococcus 

aureus with standard reference method and impedance – splitting 

method respectively , and the sensitivity of impedance method 

was calculated equal to 60 percent. The difference of methods was 

not statistically significant (P>0.05). Comparing the results in warm

and cold seasons showed that there was no contamination to 

investigated microorganisms in all 45 samples of cold season. Also 

except 5 (11.1%) butter samples which were detected as 

contaminated to Staphylococcus aureus by standard method , there 

was not any contamination to coliform and Escherichia coli among 

the 45 samples of warm season . The results showed that the 

difference of contamination rate to Staphylococcus aureus between 

cold and warm season was statistically significant (P60% of bovine strains resistant to 4-6 

antimicrobials, contrasting to human strains showing more frequent 

multidrug resistance (MDR). None of the bovine strains carried class 

1 integrons, while most (8/13) human strains did. The genes grm 

(responsible for rRNA methylase), and the genes aac-Ib, and aadB 

(encoding aminoglycoside modifying enzymes), conferring 

gentamycin resistance occurred in both groups. However, less 

frequently, and without phenotypic expression in bovine strains 

which was in contrast to human strains. Virulence determinants of the 

type III secretion system (TTSS) equally occurred in both (bovine 

and human) groups of strains. The dominant O-types were O4 (more 

typical for human) and O6 (more typical for animal) strains. Pyocine 

types, and phage types did not indicate a clear cut difference between 

the two pools. However, PFGE analysis distinguished the genotypes 

of representative bovine and human strains. Our results indicated that 

strains of Pseudomonas aeruginosa from healthy cattle have some 

traits common with human clinical strains. This may be interpreted 

only as an indirect but not a direct human health risk. It seems that the 

human clinical isolates might have evolved through several gene 

transfers (likely in a hospital environment) in order to their present 

combined pathogenic and MDR potential. 

Key words: Pseudumona aeruginosa, virulence, antibiotic resistance, 

pulse field electrophoresis, bovine and human strains 

as a rickettsia, but recently reclassified with the genus Mycoplasma 

on the basis of 16S rRNA analysis. M. wenyonii is an epicellular 

haemoparasite that attaches to red blood cells (RBC) and causes 

RBC deformity and damage, but may also be found in plasma 

unattached erythrocytes. The resulting disease, traditionally called 

bovine eperythrozoonosis, has been reported and is considered a 

problem of feeder cow, where it manifests as a parasitaemia and 

anaemia with infected cattle rarely dying but on occasions showing 

acute clinical signs. The aim of this study was performed by slide 

blood film method and polymerase chain reaction (PCR) for 

Parasitology 273

detection of M. wenyonii infection. An investigation of M. wenyonii 

was confirmed in shorthorn cows on Shandong province. Slide 

method test was carried out for the detection of Eperythrozoon in 

412 shorthorn cows from six different herds. Ninety three of the 412 

cows (22.6%) showed Eperythrozoon positive reactions, and the 

test-positive Eperythrozoon were then conformed by PCR analysis. 

81(88.2%) of the 93 Eperythrozoon positive samples revealed 

positive. Furthermore, the positive rates for M. wenyonii infection 

were 6.3% (26) in the 0-2-year-old group, 4.1% (17) in the 2-4-yearold 

group, 4.6% (19) in the 4-6-year-old group, and 4.6% (19) in the 

> 6-year-old group by PCR analysis. With regard to the age element, 

younger cows also showed a higher prevalence of Mycoplasma 

wenyonii infection than the adults in the current study, because 

statistical analysis indicated that significant difference was observed 

in

Infection of ewes by Toxoplasma gondii has serious consequences 

for sheep production, inducing abortion if the infection is acquired 

before 50 days of gestation, or fetal loss between 70 and 90 days. 

Herbivores could be used as sentinels of environmental 

contamination by T. gondii oocysts. Colima, a warm and humid state 

in the west of Mexico, presents favorable climatic conditions for 

transmission of this parasite and harbors both domestic and wild 

cats, capable of dispersing T. gondii. The aim of this study was to 

determine the frequency of specific antibodies in sheep from the 

coastal regions (South), mountains (Center and North) and Pacific 

Island of Socorro where the feline is domestic but feral. Serum 

samples from 420 sheep were tested by a previously standardized 

indirect ELISA. The frequency of infection was estimated and the 

area of origin, the age and the sex were analyzed as risk factors of 

toxoplasmosis by means of Odds ratio (95% Confidence Interval). 

Statistical significance was tested by Chi-square or Fisher’s exact 

tests. Comparison of absorbance values among groups was 

performed by one-way-ANOVA. The global frequency of anti-T. 

gondii antibodies was 26.9%. The highest levels were found in the 

coastal zone (34.3%), and the lowest on the island (15.9%), with 

older animals found to be more frequently positive and with higher 

absorbance values; all these results presented statistical significance 

(P

390 Study on Attenuation Mechanisms in Iranian Theileria 

annulata Macroschizont-infected Cell Lines 

G. Habibi 1 , S. Bozorgi 1 , K. Esmaeil-Nia 1 , R. Hashemi-Fesharki 1 , 

N. Bordbar 2 

1 Razi Institute, Protozoology, Karaj, Iran 

2 Razi Institute, Quality Control, Karaj, Iran 

Theileria annulata, the causative agent of theileriosis is tick-born 

parasitic protozoa and is highly pathogenic for cattle. Attenuated 

vaccines are an important means of controlling Theileria annulata 

infection of cattle. Production is by prolonged cultivation of 

macroschizont-infected cells. The sporozoites of Theileria annulata 

invade bovine MHC II cells, where they differentiate into schizonts. The 

latter can immortalize and induce fundamental changes in their host 

cells. The mechanisms underlying this transformation are not 

understood. In this study, three theileria annulata infected cell line were 

studied, “Sa” vaccine strain and two other theileria annulata-infected 

cell lines “C1 and C2”. The purposes of this work were to analyze that 

prolonged in vitro culture of Theileria-infected cell lines results in their 

attenuation and this process is associated with alterations in both host 

and parasite gene expression. The results demonstrated that, the infected 

cell lines show increased specific transcripts for bovine MMP9 in low 

passage cultures, but it decreased in attenuated cell lines (Sa vaccine 

strain and high passage of C1 and C2 cell lines). In addition the level of 

transcription for pro-inflammatory cytokines (IL-1 and TNF) and 

Tams1 are studied and discussed in this study. In conclusion, the results 

of this work clearly showed that the level of MMP9 transcripts are in 

contrast with the amounts of Tams1 mRNAs in T. annulata schizont 

infected cell lines for virulence and attenuation 

respectively. Understanding the mechanisms of virulence and 

attenuation of infected cell line by using molecular biology methods and 

in vivo animal experiments could help to increase our knowledge about 

attenuation mechanisms and preparing and identifying appropriate cell 

lines in order to develop the new Theileria annulata vaccine cell lines. 

Key words: Theileria annulata, matrix-metalloproteinase attenuation 

391 Evaluation of Skin DTH Theilerin Test as an Index of 

Immunity against Theileriosis after Vaccination 

K. Esmaeilnia 1 , M. Khani 2 , G. Habibi 1 , R. Hashemi-Fesharki 1 


2 Private Praxis, Karaj, Iran 

Bovine theileriosis due to Theileria annulata is the most important 

parasitic disease of cattle and an obstacle in development of animal 

husbandry economy in Iran. Vaccination against theileriosis is one of 

the most effective routes of control and prevention of the disease. 

Theileria annulata vaccine has been produced in Razi Vaccine & 

Serum Research Institute for a long period in Iran and vaccinated cattle 

show good resistance to the disease for at least 1 year. But due to many 

factors , the amount of production is not sufficient for the need of the 

country , so , evaluation of the period of protective immunity can play 

an important role in the effectiveness of this strategy in control of 

theileriosis. For this purpose dairy cows of 7 conventional farms in 

endemic region of Bouin-Zahra (Ghazvin province) were covered. Two 

methods including Theilerin (Delayed type Hypersensitivity) and 

ELISA tests were used as the indices for evaluation of cellular and 

humoral immunity , and according to a programmed time table , we 

tried to evaluate the protection in the periods of 1, 2 and 3 years after 

vaccination. The data were analyzed statistically and according to the 

results, the Theilerin skin test could be used as an index for detection of 

the level of protection in vaccinated cattle, and the sensitivity and 

specificity of this method were calculated as 88 and 31.5 %, 

respectively, in comparison with ELISA. But because of variety and 

the loss of solidity in the results the serological test was not reliable for 

this purpose, and must be replaced by another method for the future 

surveys. 

Key words: DTH test, Theileriosis, vaccination 

392 Laboratory Evaluation of Three Strains of the 

Entomopathogenic Fungus Metarhizium Anisopliae for 

Controlling Hyalomma Anatolicum Anatolicum and 

Haemaphysalis Punctata 

M. Tavassoli 1 , A. Ownag 2 , R. Meamari 1 , S. Rahmani 1 , 

K. Mardani 3 , T. Butt 3 


1 Faculty of Veterinary Medicine, Department of Pathobiology, 

Urmia, Iran 

2 Faculty of Veterinary Medicine, Department of Microbiology, 

Urmia, Iran 

3 Faculty of Veterinary Medicine, Department of Food Hygiene, 

Urmia, Iran 

Objectives of study: Hyalomma anatolicum anatolicum and 

Haemaphysalis punctata are widely distributed and harmful 

ectoparasites of livestock and vector of disease agents. The aim of this 

study was to evaluate 3 strains of M. anisopliae against H. anatolicum 

anatolicum and H. punctata. 

Materials and Methods: Three strains of M. anisopliae strains were 

used in this study. Engorged female of H. anatolicum anatolicum and 

H. punctata were collected from the naturally infested sheep and after 

sterilized in 70% ethanol transferred to Petri dishes and incubated at 

28° C and 80% RH. After laying eggs the emerged larvae’s were used 

in bioassays. The treatments were conducted by immersing larval stage 

(at least 20 larvae/Petri dish) in the spore suspension. All treated and 

untreated ticks were observed by day interval up to 18 day to detect 

dead ticks and signs of mycosis. Three replicates were made for each 

suspension. 

Results: The larval stages of H. anatolicum anatolicum and H. 

punctata were susceptible to three strains of M. anisopliae, but the 

degree of virulence of fungal strains varied considerably (p

p-nitroanilide, Ná-CBZ-L-Lysine p-Nitrophenyl Ester and 

Hemoglobin for cytoplasmic proteins and cell membrane bound 

proteins. Then, enzyme like trypsin, enzyme like chymotrypsin, 

carboxypeptidase B, cathepsin C, cathepsin B and cathepsin D were 

detected using above substrates respectively. Our results show that 

enzyme like trypsin is a membrane bound protein, but 

carboxypeptidase B, cathepsin C, cathepsin B and cathepsin D 

included in the cytoplasm of mid-gut cells . There is no enzyme like 

chymotrypsin in mid-gut cells of tick. 

Key words: Hyalomma, tick, protease, detection, assay 

429 Current Progress in the Development of Fascioliosis Vaccines 

with the Possibility of Cross Protection against 

Schistosomiasis 

A. Derbala, E. Abdel Rahman 

National Research Center, parasitology, Giza, Egypt 

Fascioliosis is an important trematode infection of herbivoera 

worldwide with increasing evidence of prevalence as a disease of 

humans.Vaccination studies with purifed native and recombinant 

Fasciola antigens suggest that this approach is a realistic goal and its 

future prospect is optimistic. Among the major potential vaccine 

candidate are fatty acid binding protein(FABP), cysteine(cathepsin) 

proteases. Haemoglobin, leucine aminopeptidase and a saposin-likr 

protein. In addition to protective effects with significnt worm burden 

reduction, some vaccine candidates also have anti-fecundity, antipathology 

and anti-embryonation effects.In the case of Fasciola 

hepatica FABP, cross- reaction and cross-protection against 

Schistosoma mansoni is an important feature. The possiblity of 

developing multipurpose vaccine for both veterinary and human use is 

a fundemental significance in terms of real progress towards effective 

immunoprophylaxsis against helminthiasis. 

Key words: Fascioliosis, Schistosomiasis, vaccine, human, animal 

430 Gastro-intestinal Nematodes Prevalence in Dairy and Meat 

Grazing Calves on Tropical Climate in Mexico 

L. Encalada 1 , M. Lopez 2 , P. Mendoza 2 , E. Liebano 2 

1 Universidad Autonoma de Campeche, Escuela Superior de Ciencias 

Agropecuarias, Escarcega, Campeche, Mexico, Mexico 

2 INIFAP, CENIDPAVET, Jiutepec, Morelos, Mexico, Mexico 

The main aims of this work was determinate the prevalence and level 

of gastrointestinal nematodes infection on grazing calves in a 

tropical climate localized in Escarcega Campeche, district of 

Mexico. Five-hundred eight Bos indicus and Bos taurus cross-breed 

calves were selected and divided in two groups (I and II), according 

to the age. The first one was studied since they were born to three 

months of age and the second one was studied from four to seven 

months-age. All calves were grazing for 12 months under tropical 

climate following all husbandry procedure, of the farm. 

Parasitological samples were analyzed each month and number of 

eggs per gram was determinate by McMaster technique. ANOVA 

analyses and the statistical test Holm-Sidak were used. Then, all 

results were analyzed following three main criteria: low infection 

when the number of eggs per gram (epg) was from 50 to 500; mild 

infection when the number of epg was from 550-1000 and high 

infection when the number of epg was >1050. The prevalence results 

showed statistical differences (P< 0.01) between groups I and II of 

(62.31%) a and (80.15%) b , respectively. On the other hand, the level 

of infection showed statistical differences (P< 0.01) determined by 

epg was around (61.40%) a for almost all the evaluated periods, 

considering this result as low level of infection, followed by 

(23.90%) b of middle infection and (14.69%) b to those which showed 

high level of gastro-intestinal epg. In addition, genus such as 

Cooperia spp, Haemonchus spp and Oesophagostomus sp were 

identified through the 12 months evaluated, although, Cooperia spp 

appeared to be the most frequent genus in calves. However, no 

association was observed between the prevalence and level in 

infection of gastrointestinal nematodes and a specific annual season 

(rain, north wind and dry seasons). In conclusion, most of the 

naturally infected calves acquired low level of infection comparing 

with the 14.69% which were severely affected by gastrointestinal 

nematodes. Also, the main nematode genus identified in Escarcega, 

Campeche as tropical region was Cooperia spp. 

Key words: calves, gastro-intestinal nematodes, tropical region 

431 Using Dot-ELISA for Detection of Experimental Babesia ovis 

Infection in Lambs 

K. Esmaeilnia 1 , R. Hashemi-Fesharki 1 , S. Nouri Ale-Agha 2 


2 Private Praxis, Tehran, Iran 

Four Splenectomized and 16 intact lambs were divided into 5 

groups, and 4 groups were inoculated with 10 7 and 10 8 B. ovis 

infected RBCs, and a group remained intact as negative controls, 

and were kept under daily observations for clinical signs of 

babesiosis including fever and hemoglobinuria. When fever 

occurred, peripheral blood smears prepared daily and after Giemsa 

staining , examined microscopically and blood sampling performed 

for serum preparation and preservation. Fever and parasitaemia, 

both occurred at 5 dpi. Parasitaemia reached to a peak of 17% at 9 

dpi and then gradually decreased. All of the splenectomized lambs, 

and also 4 lambs of other infected groups were dead during the first 

20 days of experiments, but the others survived and the experiment 

continued by them. First occurance of anti Babesia antibodies was 

at 16 days post inoculation, when the parasitemia had felt down to 

less than 1%, but the antibodies were distinguishable even after 1 

year post inoculation by this method. Somehow the IFA only 

detected these antibodies up to 9 months post inoculation. 

According to the results, Dot-ELISA is a suitable method for field 

studies on the prevalence of babesiosis and is more sensitive in 

comparison with IFA. 

Key words: Babesia ovis, Dot-ELISA, experimental infection 

432 Prevalence Study of Eimeria Species in Swedish Cattle 

A. Forslid1 , J. Dahl2 , D. Christensson3 1 Veterinary Practice, Veterinary Professional Services, Uppsala, 

Sweden 

2 Bayer Health Care, Animal Health Division, Copenhagen, Denmark 

3 National Veterinary Institute, Parasitology, Uppsala, Sweden 

Objectives of Study: No previous prevalence studies of pathogen 

Eimeria species (E. bovis and E. zuernii) in cattle have been done in 

Sweden. A study was performed in Swedish cattle herds to assess the 

prevalence of pathogen Eimeria species. 

Materials and Methods: 99 cattle farms spread all over Sweden 

were chosen for the study. Herd size varied from 30 - 980 cows. 

The study was conducted during the stable period of 2006. 541 

calves between 3 weeks and 9 months of age were sampled. The 

calves were held in groups after weaning and the samples were 

taken 3 weeks after regrouping. Faeces was collected from the 

rectum. Animals that had been treated with anticoccidials were 

excluded from the study. The laboratory examined the samples for 

presence of oocysts of Eimeria and if found positive, the oocysts 

were further speciated. A statistical evaluation was performed to 

assess, whether the observed herd prevalence was compatible with 

the assumption, that all herds were positive for either E. bovis or E. 

zuernii. 

Results: Between 1 and 12 samples were taken from 99 farms. In 

total 541 calves were sampled. 23 % of all individual samples were 

positive for either E. bovis or E. zuernii. 18 % of all calves were 

positive for E. bovis and 9 % were positive for E. zuernii. 61 % of all 

the farms had at least one individual with E. bovis and/or E. zuernii 

(55% E. bovis and 35% E. zuernii). The number of positive farms 

was dependent on the number of samples per farm. 52 % of herds 

sampled with 5 or fewer samples were positive for pathogenic 

Eimeria, while 81 % of herds sampled with more than 5 samples 

were positive. Statistical tests showed that the hypothesis, that all 

herds were positive for pathogenic Eimeria could not be 

rejected. 32% of all the farms had calves with diarrhoea at the day of 

sampling. 38 calves had diarrhoea. Calves with high levels of E. 

zuernii had a significantly higher risk of having diarrhoea at the visit. 

Calves in herds, where other calves where positive for E. bovis had 

higher risk of having diarrhoea.There was no significant difference 

in age between positive or negative calves. 

Conclusions: The study showed a high prevalence of E bovis and E 

zuernii. No significant differences were found between different parts 

of the country. The farm prevalence depended very much on the 

number of samples per farm. It is possible, that all herds are infected 

with pathogenic Eimeria species. 

Key words: Coccidia, Eimeria, cattle, diarrhoea, prevalence 

Parasitology 277


1152 Detection of Theileria Anuulata in Blood Samples of Native 

Cows by PCR and it’s Comprasion with Smear Prepration 

Method 

H. Azizi, B. Shiran, F. Salehi, Z. Sadeghi 

Veterinary Faculty of Shahrekord University, Parasitology, 


Objectives of study: In order to comparison on detection of Theileria 

annulata by PCR in blood samples from carrier native cattle’s with 

smear preparation method, in March and April 2007, 140 native cattle 

above one-year referring to slughter-hous of Najafabad was randomly 

sampled. 

Materials and Methods: For detection of parasite, blood samples 

from vein of ear were smeared and fixed with methanol, then stained 

with Gimsa dye and by light microscope with 100 x lens were 

observed. Also PCR blood samples (9 ml each) collected in tubes, 

containing 1 ml of 0.1 M, (3.2%) buffered citrate solution. 

Results: From 140 samples, 56% (40%) samples were positive and 84 

(60%) samples were negative by PCR method, whereas from 140 

samples, 12 (8.75%) samples were positive and 128 (91.43%) samples 

were negative by blood smear method. All of the samples that were 

positive by blood smear were also positive by PCR. 

Conclussion: Comparison between two methods (smear and PCR 

preparation) on detection of Theileria annulata carrier native cattles by 

M.C Namar static test showed that there is significant difference 

between these two methods. With regarded to achieved results, in this 

survey, sensitivity of PCR technique in detection of Theileria annulata 

carriers was greater than smear preparation method and superiority of 

PCR method has been demonstrated. 

1153 Study on Incidence of Blood Parasites in Slaughtered 

Camels in Tehran Abattoir 

Sh. Ranjbar-Bahadori, A. Mirzai 

Islamic Azad University, Garmsar Branch, Parasitology, Garmsar, 

Iran 

With regard to presence of one hundred-twenty thousand camels in 

different regions of Iran and using its production, also transmition of 

zoonotic diseases to other animals and even human, this study seems 

to be significant. One hundred blood samples from slaughtered 

camels in Tehran abettor were prepared to three types: with 

anticoagulant for centrifuging and isolation of Trypanosoma spp 

from their Buffy coats; with formaldehyde 2% for modified knott 

method and isolation of microfilaria; as blood slide for staining by 

gimsa for presence of protozoa. Results showed that the most 

common blood parasites in studied camels was Trypanosoma evansi 

which was seen in 6% of them. Also 4% of animals were infected to 

Anaplasma marginale and 2% of them infected to Babesia spp and in 

1% of samples were observed microfilaria of parasitic helminthes. 

Statistical analysis did not show any significant relationship between 

infection and some factors included: age and sex (P>0.05). 

Regarding the transmit ion of observed blood parasites in studied 

camels by vectors including ticks and flies, control of them can be 

effective in prevention of above-mentioned parasites. 

Key words: blood parasites, Camel, Tehran, Iran 

1154 Prevalence of Coccidia in French Young Dairy Calves in 

Vendée 

G. Denis 1 , A. Valognes 2 , A. Richard 3 

1 Cavac, La Roche Sur Yon, France 

2 Laboratoire Vétérinaire Départemental de l’Allier, Moulins, France 

3 Alpharma, Animal Health, Verrieres Le Buisson, France 

Coccidia have an underestimated economic importance in 

ruminants, and as such, there is little understanding of the extent of 

oocyst shedding. The aim of the following surveys was to gain a 

better knowledge of oocysts shedding by dairy calves during 

weaning. Fecal samples were taken from 5 to 6 calves from 4 to 10 

weeks of age across 15 dairy farms (with an average of 50 dairy 

cows) where calves which were reared in groups on straw litter did 

not show any clinical signs of coccidiosis. The laboratory used an 


iodomercurate flotation method. Identification of the different 

species of coccidia was carried out by examining the oocysts. One 

survey took place in Vendée (West France) in a dairy averaging 50 

dairy cows. In that survey, conducted in December, 2007, a total of 

76 dairy calves were tested on 15 farms. Fifty-five calves out of 76 

were positive (72 %). Only one farm with 5 fecal samples was 

negative. Eimeria bovis was observed in all farms except one, while 

Eimeria zuernii was found in 10 farms out of 15. Other species found 

included E. subspherica in 6 farms, E. alabamensis in 11 farms, E. 

subspherica in 6 farms and E. auburnensis in 5 farms. Of the 76 

samples, 21 were negative, 14 calves shed less than 100 oocysts per 

gram, 20 from 100 to 1,000 oocysts, 19 from 1,000 to 10,000, 2 from 

10,000 to 100,000 and 1 sample revealed 250,000 oocysts. None of 

these calves exhibited any signs of diaorrhea, and, as a result, none 

of them received an anticoccidial tretment. In 2002, in a survey 

carried out in Brittany, the 18 farms assessed were all positive (96 

heifers) with 8 % of the dairy calves shedding between 1,000 and 

10,000 oocysts per gram. Eimeria bovis was found on 18 farms and 

in 55 % of the calves, followed by E. alabamensis, subspherica and 

finally zuernii and auburnensis. Surveys on suckling calves were 

carried out in 2006 on 10 farms in the Charolais region using groups 

of 7 calves, aged 4 to 7 weeks. All farms were positive and the 

oocyst count was significant: 7 calves out of 70 shed between 1,000 

and 10,000 oocysts per gram of faeces, 14 shed between 10,000 and 

100,000 and 2 more than 100,000. Eimeria bovis was observed in 44 

% of the calves and Eimeria zuernii in 30 % of those sampled. This 

2007 dairy calf survey re-affirms that coccidia are present on the 

majority of farms rearing calves. 

1155 Prevalence of Stable Coccidiosis in Calves with a History of 

the Disease in the Benelux 

D. Gevaert 

Bayer SA-NV, Animal Health Division, Brussels, Belgium 

Objectives of the study: Stable coccidiosis in calves is caused by the 

coccidians Eimeria bovis or Eimeria zuernii. They are increasingly 

recognized as important protozoal pathogens in cattle. The prevalence 

of stable coccidiosis on farms has been determined under field 

conditions using faeces examinations. 

Materials and Methods: Veterinarians from Belgium, the 

Netherlands and Luxemburg submitted between May 1 st and 

November 30 th 2007 feces samples from 350 herds for screening to the 

Belgian or Dutch Animal Health State Laboratories. All participating 

herds had calves suffering from clinical coccidiosis (diarrhoea, growth 

retardation) or subclinical coccidiosis. The veterinarian collected 

individually rectally taken faeces samples of 10 gram usually from 5 up 

to 10 calves. The faeces were examined using a flotation method using 

a sugar or salt solution. The oocysts were identified as E. bovis, E. 

zuernii and E. alabamensis. All other Eimeria oocysts were registered 

as non pathogenic Eimeria spp. Upon the detection of oocysts, the calf 

was scored as positive for that species. 

Results: On 247 herds (71%) the history of stable coccidiosis was 

confirmed by detecting oocysts of E. bovis. E. zuernii also causing 

stable coccidiosis was found on 163 (46%) of the herds. On 268 herds 

(77%), oocysts of either E. bovis or E. zuernii were detected. Pasture 

coccidiosis caused by E. alabamensis was found on 186 (53%) of the 

herds whereas the non pathogenic Eimeria spp. were found on 239 

(68%) of the herds. Mixed infestations of stable and pasture coccidiosis 

were found on 167 herds (48%). 

Conclusions: These results show that in 77% of herds with signs of 

stable coccidiosis, the presence of the parasite could be confirmed. This 

number corresponds well to the 76% found in piglets for isosporosis in 

the Benelux, using a comparable methodology. It shows that the history 

of coccidiosis in a herd can be confirmed under field conditions by 

faeces examination following a standard sampling, analysis and 

identification protocol. 

Key words: Eimeria, diagnosis, prevalence 

1156 Prevalence of Sheep Liver Trematodes in the Northwest 

Region of Iran 

M. Movassagh Ghazani 1 , M. Valilou 2 

1 Islamic Azad University,Shabestar Branch, Food Hygiene, 

Shabestar, Iran 

2 Islamic Azad University,Shabestar Branch, Veterinary Pathology, 

Shabestar, Iran

The trematodes Fasciola hepatica and Dicrocoelium dendriticum are 

recognized as the most important helminthic parasite of sheep, 

causing economic losses to ranchers due to the reduction of 

production of milk and meat, condemnation of parasitized livers, 

abortions, increased mortality, and the expense of control measures. 

In Iran, the main enzootic area of sheep liver trematodes lies in the 

North of Iran, but Fasciola spp and D. dendriticum also occur in the 

Northwest region of Iran. This study was carried out to determine the 

prevalence of liver fluke infections in sheep in the northwest region 

of Iran (municipal slaughterhouse of Tabriz). For the study a total of 

140 sheep livers were collected (Systematic Random Sampling). The 

prevalence rates of Fasciola hepatica and Dicrocoelium dendriticum 

were 8.57% and 20% in sheep respectively. 

Key words: prevalence, Fasciola hepatica, Dicrocoelium dendriticum, 

sheep 

1157 Effects of Decoquinate on Bulls Growth during the First 3 

Months of Fattening: Field Trial on Charolais 

A. Richard 1 , G. Peyrin 2 , R. Jobert 3 , A. Valognes 4 

1 ALPHARMA, Animal Health Division, Verrieres Le Buisson, France 

2 Lycee Agricole Marmilhat, Clermont Ferrand, France 

3 Socaviac, Villefranche d’Allier, France 

4 LDA 03, Moulins, France 

Subclinical coccidiosis is always underestimated, it is responsible 

for poor growth. Preventing subclinical coccidiosis by group 

medication is much easier than individual treatment. Furthermore, 

daily contamination is more controlled by a daily treatment. The 

objective of this trial was to evaluate the impact of decoquinate in 

the feed on growth and feed conversion of bulls entering fattening 

units after transportation stress. Decoquinate is a non-antibiotic 

molecule active on certain protozoa: coccidia, toxoplasma, 

cryptosporidia, and neospora. It works very early in the coccidia life 

cycle at the beginning of the small intestine preventing intestinal 

damage. Decoquinate was granted a zero-withdrawal period in 

several EU countries following its classification in Annex II for 

MRL. The experiment was carried out on 117 decoquinate treated 

bulls and 58 control of 9 months of age weighing 376 kg, sourced 

from 129 different farms in the center of France. Decoquinate was 

delivered in the feed at 0,5 mg / kg BW for 28 days. Bulls were 

weighed at the end of the decoquinate treatment:11/11/2007 and 56 

days later. Oocyst sampling for count and identification was 

performed on 10 bulls in each group at the beginning of the treatment 

and at the end. The total weight gain after decoquinate treatment was 

39 kg for the decoquinate group and 34.3 kg for the control group. 56 

days after the end of the treatment, the total gain from entering the 

fattening unit was 144.8 kg for the decoquinate group and 138 kg for 

the control group: i.e., a 6,7 kg increase. The final weight were 

significantly different (P < 0,05). The feed conversion ratio for the 

duration of the treatment was 6.89 for the decoquinate group and 

7,39 for the control. For the entire feeding period, feed conversion 

was 5.35 for the decoquinate group and 5,51 for the control. It was 

noted that the decoquinate group consumed the daily ratio much 

more rapidly and had a better appearance. Oocyst levels were low in 

both groups (< 420) at the beginning of the treatment with a few E. 

bovis and zuernii recorded at the end of the 28-day period

digestion. It works only on Apicomplexa protozoans (coccidia, 

cryptosporidia, etc.). The target dosage must be always expressed as 

individual daily dosage and never as ration inclusion rate (ppm). The 

correct dose is 0,5 mg / kg BW / day with a 28 days duration 

minimum. In light of ration variation and in order to more readily 

achieve the target decoquinate dosage of 0,5 mg / kg BW a 

supplementary, pelleted, decoquinate-containing feed was designed. 

A 1,000 ppm inclusion rate was chosen so that each gram of the 

supplement delivered 1mg decoquinate. Therefore each gram of 

supplement mixed in the forage ration medicated 2 kg BW. The 

objective of this trial was to evaluate the effect of the decoquinate 

1000 ppm feed on growth and carcass weight of bulls entering 

fattening units .The experiment, conducted in fattening units, began 

in January 2006. 34 Limousin bulls, 8,5 months of age, weighing 253 

kg were utilized. 17 head received decoquinate and 17 head served 

as controls. Supplement containing decoquinate (1,000 ppm) was 

incorporated in the feed to deliver decoquinate at a rate of 1g / 2 kg 

BW/head/day for 28 days. There was no evidence of clinical 

coccidiosis during the trial. Forty-one days from study initiation, the 

decoquinate and control groups weighed 329 kg and 316 kg 

respectively. At 103 days on feed, decoquinate-fed bulls weighed 

418 kg versus 404 kg for controls. Duration of the fattening period 

was 360 days for the treated group and 381 days for the control. 

Carcass weights were 680 kg and 668 kg for the decoquinate and 

control groups respectively. This study suggests the use of 

decoquinate at a daily dosage of 1 g/2kg BW/head/day in a 

supplementary feed (1000 ppm) will improve live weight gain and 

carcass weight of bulls during the fattening period. 

1161 Effects of Imidocarb Dipropionate Use on Tripanosomosis 

by Trypanosoma vivax in Sheep 

R. Lira, F. Bezerra, J. Carvalho, H. Alves, C. Rodrigues, J. Araújo, J. 

Batista 

Universidade Federal Rural do Semi-Árido, Cincias Animais, 

Mossoró, Brazil 

The effects of imidocarb dipropionate use on tripanosomosis by 

Trypanosoma vivax in sheep were studied. Four male sheep without 

definite race were inoculated intravenously with 1ml of blood 

containing 1,25x105 trypomastigotes of T. vivax, and other 4 sheep 

were used as control. After infection, the presence of trypanosomes 

in the blood and temperature were recorded daily for a 60-day 

experiment period. Blood samples were obtained for hematological 

analysis at 5-day intervals. Fifteen days after infection, period in 

which the animals were in the acute phase and high parasitaemia, the 

treatment was made using imidocarb dipropionate (Imizol, 0,025 

ml/kg) via deep intramuscular injection. T. vivax was observed 

constantly in the blood of the inoculated sheep from 4 days after 

inoculation. The higher parasitaemia occurred between 7 and 15 

days after inoculation. Two weeks before the treatment, infected 

sheep showed anemia and fever. All animals were negative for T. 

vivax infection without signs of reinfection, from the first day 

immediately after treatment. Infected animals still presented after 

the treatment normal rectal temperature, not significantly different 

from the control group (P>0,05). The same occurred with the 

hematocrit average values that raised significantly and did not differ 

from the control group animals. The use of imidocarb dipropionate 

in the tripanosomosis by T. vivax treatment has presented 

effectiveness in promoting the disappearance of clinical signs of the 

disease and parasitaemia. 

Key words: sheep, Trypanosoma vivax, imidocarb dipropionate 

1162 Epidemiology of Neospora caninum in Dairy Cattle in 

Northeast Iran 

A. Sadrebazzaz, M. Farhoodi, F. Vahedi 

Razi Institute, Parasitology, Mashhad, Iran 

Prevalence of anti-Neospora caninum Ig G (subclass I and subclass II) 

was measured in serum samples randomly collected from 1084 dairy 

cattle from four cities (Mashhad, Neyshaboor, Torbate-heydarieh, 

Gonabad) of Northeast Iran. The presence of anti-N. caninum 

antibodies was determined by indirect immunofluorescence - antibody 

test (VMRD Inc., Pullman,WA 99163, USA) with a cut-off value of 

1:200. We randomly compared a few negative and positive samples by 

another test (Iscom Elisa). In four cities, of the 1084 samples, 178 


(16.42%) were positive. Distribution of N. caninum in four cities 

indicated range of seropositivity from 15.18% to 27.71%. The 

percentage of seropositive aborted cattle was 20.23%. The results 

revealed the presence of N. caninum infection in the herds investigated, 

drawing attention to its role as a potential cause of abortion in cattle in 

Iran. 

Key words: Neospora caninum, epidemiology, cattle, Iran 

1163 Evaluation of Anemia and Leucocytes Indexes in Cattle 

Theileriosis 

A. Ramin, S. Asri-Rezaie, M. Hemati 

Urmia University, Animal Science Department, Urmia, Iran 

Objectives: 1) the frequency of anemia in theileriosis, 2) the 

parameters influences and severity and type of anemia 3) changes in 

leucocytes. 

Procedure: 91 cows with theileriosis and 19 healthy cows as control 

group were selected. Theileriosis was confirmed by preferal blood 

smear or schizontes in LN. A 5 ml blood with EDTA was taken from 

each cow. PCV (%), Hb (mg/dl), WBC (ml), RBC (x106/ml), Ptn (g/dl) 

and absolute lymphocyte, neutrophil, eosiniphil counts (/ml), MCV 

(fl), MCH (Pg), MCHC (g/dl) and parasitemia were assessed by the 

current methods. 

Results: The mean for parameters in theileriosis were 22.1, 6.8, 

7075, 4, 6.4, 3898, 2746, 136, 55.5, 16.7 and 30.5, respectively. 

Mean concentrations for PCV, Hb, RBC, eosinophils and MCHC 

were significantly (P24, 20-23.99, 12-19.99 and 

Scotland, cryptosporidiosis appears in young calves of 5 to 15 days 

of age, with a putrid-odor mucoid diarrhea. The differential 

diagnosis is based on the presence of oocysts revealed at the lab. 

Clinical cryptosporidiosis generally appears in beef or dairy herds at 

the end of the calving period when the pressure of infection is at its 

maximum. Curative treatments in calves are few, difficult to use and 

often provide poor results. A systematic preventive treatment in 

suckling calves is also difficult to carry out owing to the constraint 

of treating them preventively once a day during 7 consecutive days. 

Moreover, weight loss is often observed. The pregnant Simmental 

cows were treated with decoquinate at the rate of 1.25 mg/kg 

BW/day over 28 days prior to calving and for 7 days following it. In 

these field trials, 120 cows were first treated during two consecutive 

pregnancies. In these farms, clinical cryptosporidiosis disappeared 

almost totally. The dosage must be strictly adhered to and the body 

weight of pregnant cows must not be underestimated (760kg in 

Simmental). Also it is important to treat before and after calving, 

cows must be grouped according to their calving date, a cow with 

delayed calving must be treated till after parturition. Our field trial 

confirms other trials already run in France and Spain on Beef cows 

and ewes. Treating cows or ewes with decoquinate 4 weeks before 

and 1 week after parturition controls cryptosporidiosis in newborn 

ruminants. 

1165 Evaluation of the Persistent Efficacy of CYDECTIN 

(Moxidectin) 10% LA Injectable against Psoroptes ovis in 

Naturally Infested Cattle in Belgium 

B. Losson1 , R. Hanosset1 , M. Lekimme1 , B. Mignon1 , E. Deroover2 1 University of Liege, Parasitology, Liege, Belgium 

2 Fort Dodge Animal Health, Animal Health Department, Brussels, 

Belgium 

The aim of the study was to evaluate the persistent efficacy of 

Cydectin (moxidectin) LA 10% injectable (Cyd. LA) in a herd of 

Belgian White and Blue cattle naturally infested with Psoroptes ovis. 

The animals were weighed, clinically and parasitologically 

examined and divided into two groups of 17 (Group 1- all positive 

for P. ovis) and 23 (Group 2 - 20 positive for P. ovis mite) animals. 

On day 0, 13 Group 1 animals and 19 Group 2 animals were given a 

subcutaneous injection of Cyd. LA at a dosage of 1.0 mg/kg bw (1 ml 

/100kg). In each group, 4 animals remained as untreated controls. 

Mite counts were performed on days 0, 28, 35, 42, 49, 56, 63, 70 and 

77 PT. Mite counts were transformed into mite scores and means 

were calculated for each group. In both pens an unexpected result 

was achieved as all experimental animals were found to be negative 

for live P. ovis mites on day 28 PT. All animals remained negative 

for live mites until day 42. On day 46 PT, two seeder animals were 

removed from Group 1 and replaced by one clinically affected and 

one clinically and parasitologically positive animal. This was not 

possible in Group 2. In Group 1, all animals remained negative for 

live mites until the end of the trial on day 77 despite the presence of 

an actively infested seeder animal; the second seeder animal was 

found positive for live mites on day 70. In Group 2, all animals 

remained negative for live P. ovis mites until the end of the trial. 

This trial indicates that Cyd. LA at a dosage of 1.0 mg/kg bw was 

highly efficient against a natural infestation with P. ovis in growing 

BWB cattle and was very well tolerated. The fact that all seeder 

animals became negative for live P. ovis mites is difficult to explain. 

Self cure can be observed but this is unlikely in all control animals. 

The high dosage linked with the use of LA moxidectin could be 

responsible for the excretion of fairly high amounts of the active 

ingredient via the urine, faces and skin secretions. The seeder 

animals could be exposed to fairly high doses of moxidectin. 

Nevertheless, in Group 1, 2 new seeders animals were introduced; 

this challenge was maintained until the end of the experiment (day 

77). All skin scrapings remained negative for live mites. In 

conclusion, Cyd. LA at a dosage of 1.0 mg/kg bw provided under 

field conditions a full protection for at least 77 days in a group of 

animals under a P. ovis challenge which was continuous from day 46 

onwards. 

Key words: Psoroptes ovis, Cydectin LA, lasting effect, cattle 

1166 Liver Copper Status in Sheep with Fascioliosis 

G. Kojouri 1 , J. Shojai 2 , I. Amani 3 

1 Shahrekord University, Clinical Sciences, Shahrekord, Iran 

2 Islamic Azad University of Shahrekord, Graduated Veterinary 

Student, Shahrekord, Iran 

3 School of Veterinary Medicine, Shahrekord University, Student, 


Fascioliosis is a disease that affects sheep, goats, cattle and many other 

species. The flukes also can migrate into the bile ducts or through 

hepatic tissue, leaving large anaerobic tracts and may produce anemia, 

hypoalbuminaemia, increasing hepatic enzyme and decreasing plasma 

ascorbic acid concentration. Liver has a major role on energy and 

protein metabolism and many important enzymes synthesis too. To this 

respect and to determine the role of parasite on copper status and also 

the role of this mineral on parasite survival, liver tissue samples were 

taken from 35 healthy and 35 infected slaughtered sheep. Copper 

concentration was measured by atomic absorption spectroscopy and 

data were statistically analyzed by ANOVA at the level of P

determined. In addition the surface of the carcase as well as the 

inside of the skin were inspected “for the detection of Hypoderma 

Larva”. In Fereydonshahr´s villages the survey was made house by 

house and in each cow after recording the sex and age, search for 

grub detection was done carefully. The species of all recovered 

Hypoderma larva were ascertained. 

Results: From the examined 294 goats in the abattoir 51.3% were 

positive. The infestation rate in three studied seasons, summer, 

autumn and winter were 39.7%, 63.8% and 60%, respectively. The 

highest (72.3%) and the lowest (35.4%) infestation rate were 

observed in December and September, respectively. Statistical 

analysis of data showed that only the differences between age groups 

of the goats were significant (P

2 AFSSA, Laboratoire d’Etudes Caprines, Niort, France 

Objective of study: Oxyclozanide is known as efficacious against the 

rumen fluke in cattle. However there are very little data about its 

efficacy in goats. The aim of this study was to check the efficacy of 

oxyclozanide in experimentally infected goats by Calicophoron 

daubneyi, both at its immature stage and adult stage. 

Material and methods: Animals included were 25 free of helminth 

dairy goats Metacercariae were obtained after artificial infection of 

Galba truncatula by miracidiums of C. daubneyi. Each goat was 

infected by 250 metacercariae on day 0. Goats were allocated in 4 

groups:-group 1: untreated control group- group 2: oxyclozanide 

(Douvistome, CEVA) at the dose of 22.5 mg/kg BW on day 10 post 

infection (PI) (immature stage: worms in small intestine) - group 3: 

oxyclozanide at 15 mg/kg BW on day 131 PI (adult stage)- group 4: 

oxyclozanide at 22.5 mg/kg BW on day 131 PI Animals were 

checked for adverse reactions following treatment. They were 

necropsied at day 138 PI. Paramphistomes were removed from the 

rumen and counted. Efficacy of the treatment was determined using 

the formula (C-T)/C x 100 (C = arithmetic mean of adult worm 

counts of the untreated control group, T = arithmetic mean of adult 

worm counts of oxyclozanide treated group). 

Results: The treatment at immature stage (group 2) with oxyclozanide 

at 22.5 mg/kg BW induced a transient diarrhea on all treated goats 

lasting less than 48 hours. No diarrhea was observed with any dosage 

applied at adult stage. The absence of diarrhea when oxyclozanide was 

applied on adult stage let us think that it might be the destruction of 

immature stages in bowel mucosae which might induce 

diarrhea.Individual establishment rates of metacercariae in the control 

group were very variable, ranging from 0 to 34% (mean = 12%).Worm 

reduction was 82% in group 2, 96% in group 3 and 96% in group 4 

compared to untreated control group. The number of worms was 

significantly lower in the 2 groups treated at adult stage than in the 

untreated control group (p

Africa and Europe, which causes morbidity and loss of productivity 

in indigenous cattle and lethal disease in imported high-grade cattle 

and crossbred stock. Samples were collected from 75 Theileria 

affected cross-bred Holstein cattle in different ages and both sexes 

and 30 healthy cross-bred Holstein cattle as control group. Serum 

proteins fractions were evaluated by cellulose acetate 

electrophoresis. The results indicated a significant difference 

between the values of total proteins of affected (37.3±18.4 g/L) and 

healthy (63.2±13.1 g/L) cattle (P< 0.005). Albumin in healthy cattle 

was 48.24±7.32 g/L and in affected cattle was 36.72±5.82 g/L. 

Globulins were increased (15.31±3.45 g/L to 23.2±4.79 g/L and 

11.9±2.91 g/L to 17.03±5.1 g/L, respectively). No difference was 

observed in globulin between the two groups. A positive correlation 

between age and serum proteins and a negative correlation between 

parasitemia rate and serum proteins was demonstrated in both 

groups. No significant differences due to sex were observed in serum 

protein concentration between two groups. 

1177 Detection of Neospora caninum DNA by PCR Analysis in 

Different Tissues of Bovine Aborted Foetuses in Infected 

Beef and Dairy Herds at Variuos Seroprevalence Level 

M. Gennero 

Istituto Zooprofilattico Sperimentale del Piemonte Liguria e Valle 

d’Aosta, Torino, Italy 

Neospora caninum is a Coccidian protozooan that in bovine species 

represent one of the most frequently diagnosed causes of abortion, 

both in dairy and beef cattle. Positivities to N. caninum has been 

signalled world-wide and also in Italy. We evaluated the presence 

of N. caninum DNA in aborted fetuses by PCR analysis in herds of 

our region in order to correlate positivity with herd seroprevalence. 

PCR analysis with Np6 and Np21 primers was carried out 71 aborted 

fetuses tissues -liver, muscle, NCS, kidney and heart- collected from 

34 selected herds (18 beef cattle herds and 16 dairy) with a prior 

history of abortion problems and seropositivity for N. caninum. 

Tested samples were considered positive when an amplified 337 bp 

specific product was present. From 21 of 34 farms of our study we 

collected blood from animals in order to test sera for N. caninum 

specific antibodies. Sera were tested with a N. caninum commercial 

ELISA-kit (HerdCheck Anti-Neospora; IDEXX laboratories). All 

tested herds had at least one positive animal to ELISA. The ELISA 

overall seroprevalence was 43%. Within-herd prevalences vary from 

5% up to 97,2%. In beef cattle we found a significantly higher 

seroprevalence than in dairies herds: 52,6% (182/243) and 36,7% 

(194/446) respectively (c 2 =62,56 and p=0,00000). Twenty seven out 

of 71 foetuses (38%) analysed by PCR were positive. Considering as 

positive foetus one with at least one PCR positive organ we found 19 

positive CNS on 27 tested (70,4%); 14 positive muscles on 

24 (58,3%) and 12 kidneys on 24 (50%). Our percentage of 38% of 

PCR positive fetuses is one of the highest descripted in similar 

surveys. Beef cattle resulted more seropositive at parturition than 

dairies ones, but in this aptitude herd we found a higher prevalence 

of positive aborted fetuses at PCR analysis. This suggests that dairy 

cattle aborted more frequently respect to beef one. This study 

confirms the importance of N. caninum as an important cause of 

abortion and that DNA detection trough PCR analysis is confirmed 

to be a valid tool for diagnosis of N. caninum infection in aborted 

fetuses. 

1178 Bovine Dyctiocaulosis: an Approach to Prevalence and Risk 

Factors in the Province of Cundinamarca (Colombia) 

O. Oliver1 , S. Barrera2 , R. Castaneda3 1 Universidad Nacional de Colombia, Animal Health, Bogota, 

Colombia 

2 Private Veterinary Practice, Bogota, Colombia 

3 Ponticificia Universidad Javeriana Colombia, Microbiología, 

Bogota, Colombia 

Dyctiocaulus viviparus causes verminuos bronchitis. It is a parasitic 

disease of the lower respiratory tract with a worldwide distribution. 

It affects mainly young animals during pasture time and is 

characterized by tracheo-bronchitis and bronchopneumonia 

depending on the animal’s immune status. This disease is observed 

in many different areas and altitudes in Colombia affecting dairy, 

beef and dual purpose cattle. Limited epidemiological studies have 


shown, using the Baermann’s technique, a prevalence between 2% 

to 43%. The objectives of this study were to determine prevalence 

and risk factors of bovine dyctiocaulosis in the province of 

Cundinamarca (Colombia) using for diagnosis the Baermann’s 

technique. A survey was done during twelve months in 22 dairy and 

dual purpose cattle farms that accepted to participate in the study. 

The statistical sample size was estimated to be 173 claves. In small 

farms all calves were sampled, in the large ones 20 calves were 

randomly sampled. The calves were 1 to 8 months old and not 

dewormed at least a month previously to sampling. The fecal 

samples were taken directly from the rectum and were evaluated 

within 24 hours using the Baermann’s technique and its sensitivity 

was improved using 30g of feces. Feces were also evaluated for 

gastrointestinal parasites using the concentration-flotation with a 

saturated solution of sodium chloride and sucrose. The interviews 

were done by one researcher. The information was to characterize 

the farm regarding size, management, feeding and health programs. 

Statistical analysis included a x 2 , Fisher exact test, Odds ratio (OR) 

and binary logistic regression (p

T. Arantes 2 , C. Buzzulini 1 , W. Lopes 1 , R. Mendonca 1 , 

C. Sakamoto 1 , R. Lima 1 , G. Oliverira 1 , A. Costa 1 

1 CPPAR/FCAV/UNESP, Animal Health Research Center, 

Jaboticabal, Brazil 

2 Intervet Schering-Plough, Desenvovimento, Sâo Paulo, Brazil 

Introduction: The aim of this study was to determine the efficacy of a 

pour on formulation containing 12% chlorpyrifos base against flies and 

ticks in naturally infested cattle. 

Materials and Methods: The study was carried out as four 

experiments. In the Exp. 1, 2 and 3, focusing on the activity against 

Haematobia irritans, 45 naturally infested animals were selected on 

the basis of the entire body hornfly count performed on D-2 and D-1 

in respect to the treatment day. The animals were treated with a 12% 

chlorpyrifos base pour on formulation (Expert, pour-on, Intervet) or 

12.5% diazinon (Neocidol, pour-on, Novartis) according to the 

producers recommendations. In each treated animal the fly count 

was performed on D3, D7 and subsequently on weekly interval post 

treatment until no therapeutic effect of the products was observed. In 

the Exp. 4, focusing on the activity against Ripicephalus (Boophilus) 

microplus, 30 animals of Holstein-Zebu cross were selected on the 

basis of three consecutive counts of female R. microplus (5.4-8.0mm 

in length) present on the left side of the animal, performed on D-3, 

D-2 and D-3 in respect to the treatment day. The animals were 

treated with a 12% chlorpyrifos base pour on formulation (Expert, 

pour-on, Intervet) or 1% fipronil (Topline, pour-on, Merial) 

according to the producers recommendations. The post-treatment 

tick count was carried out on D1, D3, D7 and subsequently on 

weekly basis until D35 post treatment. 

Results: Formulation containing 12% chlorpyrifos base showed 

higher efficacy against H. irritans than 12.5% diazinon, reaching 

100% efficacy on D3 post administration. The duration of activity 

against R. microplus of 12% chlorpyrifos pour on proved to be 

shorter than of 1% fipronil, however no difference in the efficacy for 

these two products was demonstrated until D14 post treatment. The 

efficacy of 12% chlorpyrifos against ticks was above 95% between 

D7 and D21 post administration. 

Conclusions: The results of this study indicated high efficacy of a pour 

on formulation containing 12% chlorpyrifos base against H. irritans 

flies and R. microplus ticks in naturally infested cattle. 

1181 Efficacy of 50%Chlorpyrifos + 50%Amitraz Combination 

against R. microplus in Experimentally and Naturally 

Infected Cattle 

C. Buzzulini 1 , R. Mendonca 1 , C. Sakamoto 1 , R. Lima 1 , 

W. Lopes 1 , W. Garcia 1 , G. Oliverira 1 , T. Arantes 2 , A. Costa 1 




Introduction: The aim of this study was to evaluate the efficacy of a 

formulation containing 50% chlorpyrifos + 50% amitraz against ticks 

Ripicephalus (Boophilus) microplus in experimentally and naturally 

infected cattle. 

Materials and Methods: The study was conducted as two separate 

experiments. In the Experiment 1, after the acclimatization period, 

the animals (n=10) were infected with 5,000 larvae of R. microplus, 

aged 14-21 days. The animals were then divided into two 

experimental groups (n=5, each) on the basis of the number of 

female parasites collected on D-3, -2 and -1 in respect to the 

treatment date. The animals were then treated with a combination of 

50% chlorpyrifos+50% amitraz (Amiphós, Intervet) according to 

producer’s recommendations. In the Experiment 2, twenty animals 

of Holstein breed naturally infected with the R. microplus ticks were 

divided into two groups using the count of R. microplus females 

(4.5-8.0mm in length) present on the left side of the animal on D -3, 

D-2 and D-1 in respect to the treatment date. Animals in Exp. 2 were 

treated similarly to those in Exp.1. The efficacy was measured 

through the assessment of the average number of R. microplus on the 

animals in treated and control group according to procedure 

standardized by World Association for the Advancement of 

Veterinary Parasitology (WAAVP). 

Results: In the Exp.1 efficacy above 90% was demonstrated for the 

tested formulation between D3 and D35 post treatment. At each check 

time point until D35 the efficacy of the product proved to be above 

95%. The efficacy rapidly decreased as from D35 (

Results: On D0, the animals included in both experiments showed 

no statistical difference in their initial body weight and weight gains. 

On D90 in the Exp.1, animals treated with the combination of 2.25% 

ivermectin + 1.25%abamectin showed higher gain in body weight 

than those treated with moxidectin (35.6kg vs. 31.3kg). Similarly, on 

D100 in the Exp.2, in the animals treated with 2.25% ivermectin + 

1.25% abamectin the weight gain was higher on average by 7.45kg 

than in animals treated with 10% moxidectin. In the same 

experiment on D100 no statistically significant difference was found 

in the weight gains between the group medicated with moxidectin 

and the non medicated control group. 

Conclusions: Treatment of pasture-kept Nelore calves with a 

combination of 2.25% ivermectin and 1.25% moxidectin proved to 

have positive effect on the body weight gain of medicated animals as 

compared to a non treated control group. Moreover this positive 

effect was statistically higher than the effect of 10% moxidectin 

(p>0.05). 

1184 Comparative Evaluation of Anthelmintic Action of Different 

Long Acting Avermectins in Calves and Their Effect on 

Body Weight 

W. Lopes 1 , C. Buzzulini 1 , R. Mendonca 1 , C. Sakamoto 1 , R. Lima 1 , 

T. Arantes 2 , G. Oliverira 1 , A. Costa 1 




Introduction: The aim of this study was to evaluate the efficacy of 

three long acting formulations containing 2.25% ivermectin + 1.25% 

abamectin (Solution 3,5%, LA, Intervet), 3.15% ivermectin (Ivomec ® 

Gold, Merial) and 3.5% ivermectin (Megamectin, 3.5%, Novartis) in 

the control of gastrointestinal nematodes and the effect of the treatment 

on the weight gain in calves. 

Materials and Methods: The study was performed as two separate 

experiments with 104 calves. The animals used in the study showed 

no difference in respect to the parasite load (Exp. 1) and body weight 

(Exp. 2). Exp.1: On Day 0 the animals (n=24, 6 per group) were 

treated with either of the three long acting formulations: 2.25% 

ivermectin+1.25% abamectin (Solution 3,5%, LA, Intervet), 3.15% 

ivermectin (Ivomec, Gold, Merial) or 3.5% ivermectin (Megamectin, 

3.5%, Novartis) administered subcutaneously according to 

producers recommendations. On D14 post treatment all calves were 

euthanized and necropsy was performed to assess the parasites load. 

Exp. 2: In order to monitor the changes in the weight gain in treated 

animals all calves (n=20) were individually weighted on D-30, D0 

and D60. On D0 the animals were treated as in the Exp. 2. In each 

evaluation period weight gain was recorded for each individual 

animal. 

Results: In the Exp. 1 the following nematode species were identified: 

H. placei, C. punctata, C. spatulata, C. pectinata, O. radiatum and T. 

discolor. The treatment with 2.25% ivermectin+1.25% abamectin 

showed higher anthelmintic efficacy than the remaining two 

formulations. Evaluation of body weight gains in Exp. 2 demonstrated 

the overall weight gain differences of 10.0kg, 14.5kg and 21.15kg for 

3.15% ivermectin, 3.5% ivermectin and 2.25%ivermectin+1.25% 

abamectin combination above the control group, respectively. 

Conclusions: Long acting formulation containing 2.25% ivermectin 

and 1.25% abamectin was demonstrated to have higher anthelmintic 

efficacy than 3.15% ivermectin or 3.5% ivermectin and lead to 

higher weight gain in treated calves. All treatments groups showed 

significant weight gain in comparison to non treated control animals. 

1185 Anthelmintic Activity and Effect on Body Weight of a 2.25% 

Ivermectin + 1.25% Abamectin Combination in Comparison 

to other Endectocides in Calves 

R. Mendonca 1 , C. Sakamoto 1 , R. Lima 1 , W. Lopes 1 , C. Buzzulini 1 , 

H. Silva 1 , G. Oliverira 1 , T. Arantes 2 , A. Costa 1 




Introduction: The aim of this study was to compare the 

anthelmintic efficacy and effect on body weight of a formulation 

containing 2.25 %ivermectin + 1.25% abamectin and other 

endectocides in cattle. 


Materials and Methods: The study was conducted in Brazil as two 

experiments with a total number of 84 calves naturally infected with 

gastrointestinal parasites. In the Exp. 1, the animals (n=24, 6 per 

group) were treated with three long acting formulations: 2.25% 

ivermectin + 1.25% abamectin (Solution 3.5% LA, Intervet), 3.15% 

ivermectin (Ivomec, Gold, Merial) and 3.5% ivermectin 

(Megamectin 3.5%, Novartis) according to the producers 

recommendations. On D14 post treatment all the calves were 

euthanized and the parasitic load was assessed at necropsy. In the 

Exp. 2, the animals (n=60, 15 per group) were administered either 

with 2.25% ivermectin + 1.25% abamectin (Solution 3.5% LA, 

Intervet) or 1% moxidectin (Cydectin 1%, Fort Dodge) or 1% 

doramectin (Dectomax 1%, Pfizer). Individual body weight 

measures were conducted for all animals on D-30, D0 (treatment 

day), D60 and D120 in respect to the time of treatment. 

Results: In the Exp. 1 no statistically significant difference was 

found between the calves treated with three tested products in 

respect to the presence of: H. similis, C. pectinata, T. axei, T. 

colubribormis, O. radiatum, O. ostertagii and O. lyrata. The 

formulation containing 2.25% ivermectin + 1.25% abamectin was 

however statistically more effective than the other two products 

against H. placei and C. punctata, the two most prevalent species in 

Brazil. In the Exp.2 calves treated with the product containing 2.25% 

ivermectin + 1.25% abamectin showed statistically significant 

advantage as compared to the animals that received 1% moxidectin 

(weight gain difference 12.4kg) and 1% doramectin (weight gain 

difference 12.54kg). 

Conclusions: The results of the study indicated that formulation of 

2.25% ivermectin + 1.25% abamectin showed significantly higher 

activity against H. placei and C. punctata than 3.15% or 3.5% 

ivermectin. Moreover medication with the first formulation resulted in 

superior gains in body weight as compared to 1% moxidectin or 1% 

doramectin. 

1186 Efficacy of an Avermectin Association against 

Gastrointestinal Helminths Resistant to Ivermectin and 

Doramectin in Cattle 

V. Rangel 1 , R. Leite 1 , G. Almada 2 , T. Arantes 3 , J. Costa 1 , 

E. Santos 4 , P. Oliveira 1 

1 UFMG, Dept. Escola de Veterinária, Belo Horizonte, Brazil 

2 UVV, Dept. Bioagrárias, Vila Velha, Brazil 



Introduction: Anthelmintic resistance is becoming a serious 

problem in cattle world wide, especially in countries like Brazil. The 

aim of this study was to assess the efficacy of an avermectin 

association against gastrointestinal helminths resistant to ivermectin 

and doramectin. 

Materials and Methods: The study was conducted in Brazil on a 

beef cattle farm with reported lack of efficacy for ivermectin and 

doramectin. Fecal egg reduction test confirmed low efficacy of these 

substances, but not of abamectin. Egg samples were used to produce 

larvae for controlled infection. Calves (n=54) were infected with 

larvae of H. placei, C. punctata and O. radiatum (dose: 11.000, 

11.000 and 48.000 larvae respectively). After 46 days, 48 calves 

were selected and evenly distributed into 8 groups. Four treatments 

were used, 2 groups per treatment: T1 - Control group, untreated, T2 

- doramectin 1% (Dectomax, Pfizer), T3 - ivermectin 3,15% 

(Ivomec, Gold, Merial) and T4 - ivermectin 2.25% + abamectin 

1.25% (Solution 3.5% LA, Intervet). The products were 

administered according to the manufacturers recommendations. 

Necropsies were conducted to determine the efficacy of the 

treatments. In order to evaluate the persistence of efficacy, 4 

experimental groups were evaluated at 14d and other 4 groups at 28d 

after treatment. The study was carried out according to the 

recommendations of the World Association for the Advancement of 

Veterinary Parasitology (WAAVP). 

Results: The efficacy of the treatments at 14d post administration 

and calculated as geometric mean was T2: 73%, 74% and 0%; T3: 

24%, 37% and 0%; and T4: 67%, 96% and 86%, against H. placei, C. 

punctata and O. radiatum, respectively. The 28d efficacy was T2: 

38%, 79% and 16%; T3: 44%, 65% and 0%; and T4: 90%, 97% and 

94%, against H. placei, C. punctata and O. radiatum, respectively. 

Conclusions: The results demonstrated that strains of H. placei, C. 

punctata and O. radiatum resistant to ivermectin and doramectin

emained susceptible to the combination of 2.25% ivermectin and 

1.25% abamectin. 

1187 Effect of Biochemicla Blood Serum (ALT, AST, ALP, LDH, 

CA, P, CL) in Theileriosis Disease 

A. Mohagheghzadeh 1 , B. Fallahi 2 

1 Islamic Azad University, Faculty of Veterinary Medicine, Veterinary 

Department, Kahnooj Branch, Kerman, Iran 

1 Veterinary Clinic, Shiraz, Iran 

Theileriosis in cattle is caused by protozoon named as theileria 

annulata and the symptoms of diseases are by body temprature 

anemia which leads to death. In this study the seruma was separated 

from the blood of those 20 cases which were confirmed as infected 

by using blood smear test and lymphnode trephining and clinical 

test. Again blood samples were taken from 20 clinically comfirmed 

health case and seruma was separated,then the rate of following 

biochemical factors in the seruma was measured the result show that 

the activity of afew seruma enzymes such as alt,ast,alp,ldh increase 

in sick case in comparison with healthy ones (p0.05) 

1188 The Study of Culicoides Spp. as Probable Vectors of 

Bluetongue Virus to Ruminants in Western Azarbaijan, 

Eastern Aza., Ardbil and Zanjan Provinces of Iran 

M. Abdigoudarzi 1 , R. Kargar 1 , Z. Ghamari 2 , B. Razmarai 1 

1 Razi Institute, Dept. Parasitology, Karaj, Tehran, Iran 

2 Iran Veterinary Organization, Clinical Sciences Department, 

Ormoz, Iran 

Bluetongue is a viral disease that is transmitted by Hematophagus 

Culicoides spp. Domestic and wild animals are at risk, but acute 

symptoms could be seen just in sheep. A serological survey on 

Iranian sheep has been done in 1988 and twenty six different 

culicoides spp. have been reported in 1968 from different 

Geographical places of Iran. North- western provinces of Iran 

(Eastern Azarbaijan, western Azar., Ardbil and Zanjan) have been 

selected for collection of culicoides specimens. Specimens have 

been collected during night by a light trap. Collected specimens were 

put in a refrigerator, then transported in cold vessels to the 

laboratory. After primary identification of culicoides spp., some of 

them were sent for molecular study and injection to susceptible 

sheep. Samples from twenty five different regions have been 

collected during summer 2004 to 2006. Culicoides spp. From ten 

different regions were sent to reference laboratories in Germany and 

Poland. Finally, nine different culicoides species were verified. 

Molecular studies failed for virus confirmation after treatment of 

two different samples, and there were no viral growth and disease 

confirmation after injection of extract from pooled culicoides spp. to 

sheep. Additional field studies and more collections of different 

Culicoides spp. is needed for final confirmation of the role of 

Culicoides spp. as Bluetongue vectors in Iran. 

1189 A Method for DNA Isolation for Polymerase Chain Reaction 

Detection of Ornitobilharzia turkestanicum 

(Schistosomatidae) in Infected Snails 

GH. Motamedi 1 , A. Ghoreshi 2 , H. Dalimi 1 , H. Paykari 1 , 

R. Salehitabar 2 , GH. Karimi 1 , M. Abdigoudarzi 1 

1 Razi Institute, Parasitology, Karaj, Iran 

2 Institute of Agricultural Biology and Biotechnology (IBBA), 

Department of biology and genetics, Tehran, Iran 

Schistosome blood flukes parasitize mammals and birds. 

Ornithobilharzia turkestanicum (OT) as a member of 

Schistosomatidae family can infect domestic and wild 

ruminants.Since intermediate hosts of these species may be found in 

the same freshwater habitat and have morphologically similar 

cercaria. Accordingly better means are needed to take them apart. In 

the present study, a polymerase chain reaction (PCR) assay has been 

developed, for detection of OT DNA in infected animals. Specific 

primers were designed based on published OT genome sequences. 

The DNA was extracted from adult parasites and PCR was optimized 

subsequently. A 724 bp DNA fragment of the correct size was 

amplified using DNA from a single worm. PCR product was cloned 

in a pTZ57R/T vector and sequenced. Specificity of the PCR product 

was confirmed by sequencing results. All soft tissues of snails, were 

used for DNA extraction From parasite nested-PCR has been used by 

one pair of new primers. And amplification using this primers pair 

derived a 449 bp product in infected snail. Identification of an 

infected snail in pooled with aliquots from several uninfected snails 

could increase the feasibility of using nested-PCR assay using for 

detection of the hidden infection in a colony of snails.The results 

demonstrate that PCR and nested-PCR assays represent an excellent 

alternative or additional tools for the detection of OT in infected 

samples. 

Key words: plathyhelminthes, Ornithobilharzia turkestanicum, PCR, 

nested-PCR 

1190 Detection of Echinococcus granulosus in the Stray Dogs: 

Coprodiagnosis ELISA as an Alternative to Necropsy 

Gh. Motamedi 1 , M. Syavashi 2 , H. Dalimi 1 , M. Mohammadi 1 , 

S. Solymani 1 

1 Razi Institute, Parasitology, Karaj, Iran 

2 Pasteur Institute, Parasitology, Tehran, Iran 

Cystic echinococcosis has been recognized as major public health 

problem in sheeprising regions of the world. Different studies have 

shown that echinococcosis is main health problem among human and 

domestic livestock in several provinces of Iran. The development of 

sensitive and specific diagnostic techniques for detection of 

echinococcosis in humans, dogs and livestock is important for 

carrying out field studies and control programs. Necropsy of dogs 

and examination of the small intestine for the detection of intestinal 

infections with Echinoccoccus granulosus, is laborious and ethically 

questionable procedure and is not suitable for mass screening. 

Currently the most practical approach for diagnosis of E. g. in dogs 

is the method which by that antigens of parasite in faecal samples by 

ELISA using antibodies against E/S antigens from proglottids, adult 

somatic and protoscolex products are detected. This method 

improves both diagnostic specificity and sensitivity and is showed 

usfulness for epidemiological studies. Fertile sheep hydatid cyst was 

collected from sheep at slaughterhouse during meat inspection. Two 

dogs were infected orally by eating fertile hydatid cysts.Fecal 

samples were collected during periods of 1998-2003 from 83 dogs, 

shot by gun or dead by accident. Somatic (worm) antigen was 

prepared from non gravid adult E. g. worms obtained from 2 

experimentally infected dogs. ELISA for detection of parasite 

proglottis somatic products (So-Ag)in faecal samples(coproantigen) 

were performed. The presence of E.g. were recorded in 36 (43%) of 

intestine contents after Laboratory examination. The worms burdens 

were between 33-4322 E.g. The specificity and sensitivity were 80% 

and 83% respectively. Negative and positive predictive value were 

85% and 76% respectively. 

Key word: Echinucoccus granulosus, echinococcosis, ELISA, 

coproantigen 

1191 Homeopathy in the Control of Natural Infestation of 

Boophilus microplus in Cattle 

M. Arenales1 , N. Silva2 , J. Moletta2 , A. Minho2 , L. Filippsen2 1 Instituto Brasileiro De Homeopatia Veterinaria, Research, Sâo 

Paulo, Brazil 

2 Instituto Agronômico do Paraná, Animal Health, Ponta Grossa, 

Brazil 

The use of homeopathic and isotherapeutic medicines promises to be 

an alternative for the control of ticks, because they do not have reports 

of resistance of pathogens to the homeopathic therapy, and do not 

generate toxic residues to the environment. This work had as objective 

to test the use of the homeopathic product FACTOR C&MC * , for the 

control of B. microplus in heifers of the Puruna breed (breed developed 

by the Instituto Agronomico do Paraná), at their experimental farm in 

Ponta Grossa, State of Paraná, Brazil. 

Material And Methods: Twenty seven heifers aging 12 to 14 months 

were assigned to three groups for the evaluation of the homeopathic 

treatment: GI as not treated control, GII as control treated with Amitraz 

and GIII treated with FACTOR C&MC (isotherapeutic). All animals 

were maintained under field conditions of pasture and supplemented 

with 1.5 kg of concentrated (corn and soybeans) once a day, with the 

Parasitology 287

addition of mineral salt, during 12 months. Group GI did not receive 

any treatment against ticks. The animals of group GII were treated four 

times (dips) with Amitraz (1 liter per 1000 liters of water), on strategic 

days (during warmer times of the year), with a minimum interval of 21 

days between treatments. Group GIII received the homeopathic 

product FACTOR C&MC, previously mixed with common mineral 

salt (400 g in 25 kg). The amount of 100 g/day of that mixture was 

supplied mixed to the concentrate, corresponding to the dose of 1.6 g of 

product/animal/day. The weight of the animals was evaluated monthly 

and the counting of engorged females of B. microplus, was performed 

every 14 days. 

Results: The animals of the group treated with the homeopathic 

product presented a reduction of 50.5 % in the average number of 

engorged females counted during the experimental period, in relation 

to the not treated control (P < 0.05). There was no significant difference 

between the groups treated either with Amitraz or the homeopathic 

product (P > 0.05). 

Conclussion: The use of the homeopathic product showed to be an 

equally efficient alternative for the B. microplus control, mainly when 

targeting for ORGANIC livestock Production. 

* Trade mark of Arenales Flora & Fauna Homeopatia Veterinaria 

Key words: organic, homeopathy, ticks, residues, resistance 

1192 Adjuvant Effect of LPS and Propionibacterium acnes 

Combined with Anthelminthic during Experimental 

Nematode Infection in Sheep 

P. Denapoli 1 , S. Chen 1 , C. Garcia 1 , F. Toniazzi 1 , J. Mussalem 2 , 

C. Squaiella 2 , I. Longo-Maugeri 2 , R. Shaw 3 , L. Abel 1 

1 Sao Paulo University - UNIP, Departamento de Medicina 

Veterinaria, Sâo Paulo, Brazil 

2 UNIFESP Universidade Federala de Sao Paulo, Departamento de 

Medicina Veterinaria, Sao Paulo, Brazil 

3 AgResearch Limited, Wallaceville Animal Research Centre, Upper 

Hutt, New Zealand 

Introduction: Among the problems that affect sheep breeding, 

gastrointestinal helminthiasis is of primary concern. The emergence of 

anthelmintic resistance in sheep has led to seek new therapeutic 

alternatives to control helminthiasis in sheep. 

Objectives of study: The purpose of this study was to determine the 

effectiveness of an immunostimulant combined with anthelmintic on 

experimental helminthiasis through IgE analysis, T cell proliferation 

assay, haematological examinations and fecal egg counts (FEC). 

Materials and Methods: Crossbred 12-month-old male sheep were 

grouped into 4 groups: I- animals treated with LPS and 

Propionibacterium acnes and anthelminthic; II- animals treated with 

Propionibacterium acnes and anthelminthic; III- animals treated with 

only anthelmintic; and IV- animals treated with a sterile saline solution 

0.9%, as untreated controls. All the animals received 10,000 infective 

larvae (L3) orally on day 0 and were treated when the fecal egg counts 

peaked at 35 days; that is, the treatment was administered on the 35 th , 

63 rd and 67 th days after infection. Fecal and blood samples were 

collected weekly for fecal egg counts (FEC), IgE antibodies detection, 

haematological examinations and T cell proliferation assay. 

Results: T lymphocytes stimulation indexes in the presence of Con-A 

were significantly higher on the 48 th and 63 rd in the animals from group 

II. During the treatment, the animals from group I and II showed higher 

IgE antibody levels in serum than other groups (p

and acute angiocolitis also hemorrhagic-necrotic paths, a previous 

aggressions expressed by chronic angiocolitis and hepatitis. These 

modifications are characteristic for the traumatic hepatitis produced 

by Cysticercus tenuicollis (cysticercosa hepatitis) during the 

hepatical migration, extremely aggressive, metacestod identified 

along with other parasitical structures disposed in the hepatical 

tissue.Multiple and combineted parasitic aggression against body of 

Black Goat species (Rupicapra rupicapra) was complex, and for a 

long time, determining a local reactivity that was finally outnumber 

by their action on parasitic species. 

Key words: black goat, parasitic aggression, reactivity 

1195 Comparison of Oral Drench vs. Top-feeding as Anthelmintic 

Treatment in Fattening Bulls 

P. Houffschmitt 1 , Y. Lagalisse 1 , C. Mage 2 

1 Intervet Schering-Plough, Livestock Business Unit, Angers, France 

2 Animal Health Consultant, Private Veterinary Practice, Pierre- 

Buffiere, France 

Objectives: Fenbendazole (FBZ) as a 10% w/v suspension (Panacur) 

must be delivered per os. It can be achieved either by oral 

administration with a drenching gun or by superficial application on the 

feed (also called “top-feeding”). This monocentric randomised and 

controlled field study with natural nematode infestation compares the 

efficacy of both ways to administer FBZ, through the classical Faecal 

Egg Count (FEC) Reduction Test (FECRT). 

Methods: Charolais bulls (n=93) aged 7 to 9 months (mean body 

weight 166 kg), were faecal sampled on D-10. They had grazed with 

their dams and had received no anthelmintic treatment before. Bulls 

were housed and randomly allocated to 3 groups on D0, equilibrated 

for their D-10 FEC. All were fed daily with concentrate at the neck 

yoke. Group A (n=31) and Group B (n=31) were weighed individually 

on D0 and received orally 7.5 mg/kg FBZ either with a drenching gun 

or by superficial application on the concentrate feed. They were kept in 

confinement for several hours to ensure total consumption of the 

supplemented feed. Group C (n=31) received no treatment (control 

group). All 3 groups were kept in the same building and fed the same 

regimen. All animals were faecal sampled on D7. Faecal samples were 

analyzed at LDA Limoges (modified Mac Master method, sensitivity 

of 15 eggs per gram (EPG)). 

Results: Eggs found on D-10 were mainly of gastro-intestinal 

nematodes (GIN), 65% of which belonged to Cooperia sp and 35% 

Ostertagia sp. A few bulls expelled Eimeria sp cysts (n=19), 

Nematodirus sp (n=6) or Trichuris sp eggs (n=15). On D-10, the 

difference of nematode FEC between groups was not significant 

(p

digestion could be improved with improved fodder use. The footprint 

of our cows is actually very small if you consider that they are 

vegetarians and even allowing for a 12 times greater affect of methane 

on global warming than carbon dioxide, the only cost to the 

environment is the delivery of the products such as meat and dairy to 

the folks in the city. 


1231 Variations of the Antioxidant Systems in Blood of the 

Lactating Dairy Cows Exposed to Solar Radiation 

D. Pusta 

USAMV Cluj-Napoca, Animal Welfare, Cluj-Napoca, Romania 

The objective of the study is to assess whether hot season affects the 

antioxidant enzymes level (superoxide dismutases - SOD, 

glutathione peroxidase and catalase) in blood of lactating dairy 

cows. 

The study was made in the period May - August 2006, on a group of 20 

Romanian Simmental dairy cows, during the 3-rd and 4-th lactation, 

clinically healthy, kept on pasture during the day and in stable during 

the night. The determination of SOD was made by Nitro Blue 

Tetrazolium Method (NBT Method), the catalase was determined using 

photometric method with K 2Cr 2O 7 and the glutathione peroxidase 

activity was determined by guaiacol method. There were also measured 

the values of the main meteorological parameters, such as: air 

temperature, humidity and solar radiation, provided by an Adcon 

Telemetry Station. Using the measured air temperatures and humidity 

there were calculated the Temperature Humidity Indexes (THI), by 

classical methods. The mean daily THI was 73.8 ± 2.3 in August 

compared to 63.2 ± 4.6 in May. The results of the experiment showed 

that during the hot summer days, when THI is higher than 72, which is 

considered to be the superior limit of thermal comfort in cows, the 

oxidative stress appears in lactating milking cows. This oxidative stress 

is mainly manifested by the increasing of SOD with 115% (p

manufacture, roughage production, electricity utilization, 

transportation and organic waste handling were included in the 

system boundaries. Two scenarios, the system with biogas 

production and the one in which manure was applied to plant 

directly were compared. Global warming impact was offset in the 

scenario that produced biogas. Although direct application of 

manure as fertilizer could avoid the impact from chemical fertilizer 

production, offset was not be able to achieve. Organic waste 

treatment by an anaerobic digestion and recovery of energy in the 

form of biogas were proved to be useful for global warming 

mitigation from beef meat production in the country. 

Key words: LCA, Thailand, beef, meat, global warming 

1267 A Case Report from Congenital Abnormality in a Lamb 

with Brachygnathia and Cleft Palate 

Rezazadeh F. 1* , Hamali H. 1 , Ashrafi Halan J. 1 

1 Department of Clinical Sciences, Faculty of Veterinary Medicine, 

University of Tabriz, Tabriz- Iran. 

2 Department of Pathobiology, Faculty of Veterinary Medicine, 

University of Tabriz, Tabriz- Iran. 

Abstract 

A mail lamb was born in mixed sheep and goat (with 650 head) flock in 

the suburb of Tabriz (Hervy) on January 11, 2007. The breed of this 

lamb was Gezel and showed brachygnathia and Cleft palate. Lambing 

was associated with dystocia. At physical examination, lamb was 2 

days old. The dam had not anomalies in previous lambing (according to 

client speech). Macroscopic examination presents the case had not 

palate (hard and soft) and its length from nostril to commissure of the 

lip measured 11 cm. In maxilla there was an opening with 1.5 cm in 

width and 8 cm in length. The mandible bone had not any connection 

with temporal bones joint in both sides. Analysis of urine and blood 

samples did not present any significant variation expect a slight 

lymphopenia (2580/µl). Feeding of this flock was on pasture and 

occasionally some wheat silage, bread and bran. It is suspected that 


1197 The Effect of Subclinical Experimental Infection with 

Haemonchus Contortus on Carcass Quality of Karadi 

Lambs 

N. Jakhsi, M. Rekani 

College of Veterinary Medicine, University of Dohuk, Dept. of Animal 

production,, Dohuk, Iraq 

The current study was conducted to investigate the effect of sub 

clinical infection with Haemonchus contortus on physical properties 

of karadi lambs carcasses. Atotal of 14 karadi lambs 5-7 months old 

were used in this study. The lambs were divided into two groups. 

The first group (7 lambs) was considered as a control and remained 

uninfected .The second group (7 lambs) were infected with 500 

infective larvae (L3) of Haemonchus contortus Kg -1 body weight. 

This study continued until 1 th weeks post-infection & at the end of 

the study all lambs were slaughtered worm burden carcass quality 

were estimated. Data were statistically analyzed by spss program 

using T-test to determine the differences between control and 

infected group. The result revealed that there was differences in 

mean live weight at slaughter, empty live weight, hot carcass 

weight, chilled carcass weight and dressing proportion were 

(52.71),(48.64); (47.99),(43.78); (27.65),(24.62) ; (26.99),(24.03) 

and (51.16%), (49.08%) between control and infected group 

respectively but these differences statistically was not significant 

However there was slight differences in rib-eye muscle area 

(Longissimus dorsi) and fat thickness (mm) between control & 

infected group (13.94),(13.41) and (7.15),(5.32) mm respectively but 

these differences statistically not significant. No significant 

difference were found in physical composition of carcasses in leg, 

fore shank and three ribs (9,10,11) area between control & infected 

multiple genes cause congenital anomaly in skeletal system and to 

distinguish a single gene is very difficult especially when the clinical 

case is very few for the study. Teratogenic factor such as toxic plant is 

one of major etiologies in congenital anomalies that always should be 

considered. 

Key words: brachygnathia, congenital anomalies, cleft palate, lamb. 

1268 Seroprevalence of Q Fever in Selected Populations of Dairy 

Cattle in Turkey 

Gazyagci S. 1 , Aktas M. S. 2 , Kilic S. 3 , Babur C. 3 , Celebi B. 3 , Duru S. Y. 1 

1 University of Kirikkale, Department of Internal Medicine, Kirikkale, 

Turkey 

2 University of Ataturk, Faculty of Veterinary, Department of Internal 

Medicine, Erzurum, Turkey 

3 National Research Center, Epidemiology and public health, Ankara, 

Turkey 


Turkey 


Turkey 

6 University of Kirikkale, Department of Internal Medicine, Kirikkale, 

Turkey 

Animal Hygiene, Housing and Biosecurity 

Coxiella burnetii causes Q fever in humans and animals (Coxiellosis) 

worldwide. The prevalence of Coxiella burnetii antibodies in cattle has 

been investigated. 322 blood samples were collected from cattle. The 

serum was separated 24 h after sampling and stored at –20°C until 

tested. The presence of IgG antibodies to Coxiella burnetii was 

measured by indirect immunofluorescent antibody technique (IFA). 

IFAT was performed according to the instructions of the manufacturer. 

Serial twofold dilutions of positive sera were tested to determine end 

titres. Cattle with 1:32 phase II antibodies were considered as positive. 

40 out of 322 bovine serum samples (12,4%) were seropositive in the 

IFA test. 

Key words: cattle, IFA test, Q fever 

groups. Generally the results indicate that there was slight effect of 

subclinical infection with H. contortus on the productive 

Performance of Karadi lambs. The study also revealed that there was 

variation in resistance and resilience of lambs or there susceptibility 

to parasitic infection. 

1198 Airborne Dust and Endotoxin in Dairy Cows Stable 

M. Vucemilo 1 , K. Matkovic 1 , B. Vinkovic 2 , V. Vucevac Bajt 3 , 

S. Radovic 4 , R. Brezak 2 , M. Benic 5 

1 

Veterinary Faculty Zagreb, Department of Animal Hygiene, 

Environment and Ethology, Zagreb, Croatia 

2 

Croatian Veterinary Institute, Department of Ecology, Zagreb, 

Croatia 

3 

Veterinary Faculty Zagreb, Department of Humanities, Zagreb, 

Croatia 

4 

Ministry of Environmental Protection, Physical Planning and 

Construction, Environmental Protection, Zagreb, Croatia 

5 

Adria group, Adria servis, Zagreb, Croatia 

Airborne dust and endotoxins in animal houses are significant 

pollutants with a potential health risk for animals and humans 

working in these facilities. Study assessing the level of airborne 

contamination in dairy cows stable. The content of airborne dust and 

endotoxin was determined. The study was conducted in a dairy 

stable, dimensions 17.00x14.00x2.5 m. During the study, there were 

25 Simmental lactating cows and 5 calves in the 

stable. Measurements were done in the morning (before feeding and 

milking) and in the evening, once a week, during two winter months. 

Measurements were performed in the stable, in the area of animal 

stay along the feedlot. Air temperature (°C), relative humidity (%) 

and airflow rate (m/s) were determined by use of a TESTO device 

(Testo Inc., Germany). Dust was sampled onto filters (Whatman 

International Ltd., Maidstone, UK) by use of an SKC pump (SKC 

Animal Hygiene, Housing and Biosecurity 291

Ltd., Blandford Forum, UK). The airflow rate was 4.0 L/min. Filters 

were weighed before and after sampling in a controlled laboratory at 

air temperature of 22 °C and relative humidity of 45% (5%). Air 

samples were stored at -20 °C for 2-3 months until endotoxin 

analysis. Endotoxin was analyzed using the end-point Limulus 

amebocyte lysate method (LAL).The measured parameter values 

were processed by use of Microsoft Excel and Statistica 6 software. 

The analysis included descriptive statistics and statistical 

significance set at 1% (p

395 Development of a Program for Disaster Situations and 

Animal Health Emergencies in the X th Region of Chile 

G. Monti, G. Ramirez, M. Amtmann 

Universidad Austral de Chile, Institute Preventive Veterinary 

Medicine, Valdivia, Chile 

Objectives of study: It aimed to aware farmers, practitioners and 

authorities on the need and strategic importance of develop for animals, 

a comprehensive emergency plan or program for disaster situations 

(natural or man-made) or animal health emergencies and to mitigate 

physical and economical consequences of calamities though disaster 

preparedness. 

Materials and Methods: The team consists in a Veterinary 

Epidemiologist, a Rural development and technology transfer specialist 

and a Public Officer specialized in Public Health. The target audience 

was farmers, practitioners, governmental authorities, veterinary 

students and NGO aid-organizations of the X th region of Chile. The 

team tackled the problem by using an approach known as social 

marketing (defined asIt consist in participative discussion through 

creation of mechanisms that integrate all participants (including social 

organizations) for the strategic design of a preparedness plan, with 

indicative documents that facilitate the processes that incorporate risk 

or uncertainty in the planning of such plan. Expert eliciting techniques 

were also used. 

Results: Several meetings with different stakeholders (farmers, 

practitioners, governmental authorities, military and police forces, 

veterinary students and NGO aid-organizations) took place. 

Farmer’s perception of risk was focused on different issues in 

comparison with governmental authorities.Chilean society is 

undergoing changes in its attitude toward animals. Two of the most 

important changes are the urban’s society’s increasing acceptance of 

companion animals as family members and the increasing 

productivity of the animal production units in rural areas. Concurrent 

with this change, there is a growing appreciation of the value of the 

human-animal bond, improved food safety and sustainability aspects 

(conservation of the environment, land use and quality of life. 

Veterinary disaster preparedness interfaces with all of these issues 

because they all can become threatened and disrupted in disasters. 

Conclusions: The project was constructed on the existing knowledge 

development of mitigating disaster focused in a cluster of 

multidisciplinary team that together contribute towards achieving a 

common problem-oriented objective. 

Key words: preparedness, disasters, community 

396 Flunixin Meglumine (Finadyne ® , Banamine ® ) in Pain 

Management of Cattle: Pharmacology and Applications 

A. Skidmore 1 , J. Roder 1 , J. Van De Ven 2 , P. Cloet 3 , A. Montgomery 4 

1 

Schering Plough Animal Health, Livestock Technical Services, 

Canyon, United States 

2 

Schering Plough Animal Health, Brussels, Belgium 

3 

Schering Plough Animal Health, Levallois-Perret Cedex, France 

4 

Schering Plough Animal Health, Harefield, United Kingdom 

Flunixin meglumine (Finadyne ® , Banamine ® ) is a potent, nonsteroidal, 

non-narcotic analgesic with anti-inflammatory, antiendotoxic 

and anti-pyretic properties for use in cattle. Veterinary 

products containing flunixin are widely registered for cattle and 

widely used by veterinarians. In cattle, flunixin (Finadyne ® 

Solution) is indicated for the control of acute inflammation 

associated with respiratory disease. It has also been shown to have 

some benefit in the treatment of experimental acute bovine 

pulmonary emphysema (Fog Fever). Flunixin also may be used as 

adjunctive therapy in the treatment of acute mastitis in dairy 

cows. The pharmacology of flunixin is important to understand prior 

to therapeutic use in pain management. Flunixin is a weak acid 

(pka=5.82) and is highly protein bond to plasma proteins in cattle (> 

99%). After intravenous administration, flunixin (2.2 mg/kg) had a 

peak plasma concentration of 16.16 ± 5.28 mcg/ml. The unbound 

(free) flunixin is widely distributed throughout the body and exhibits 

a long elimination half life (6.87 ± 0.49 hours). In horses, flunixin 

persists within inflamed tissues and is responsible for anti- 

Miscellaneous 

inflammatory properties well beyond the time of detectable plasma 

concentrations. In calves, flunixin administration (2.2 mg/kg i.v.) 

resulted in an area under the curve (AUC) of 11.8 µg / h mL -1 and 

27.6 µg / h mL -1 in the plasma and acute inflammatory exudate, 

respectively. This gives an exudate to plasma ratio of 2.3 for flunixin 

indicating greater penetration of the drug in the inflammatory 

tissues. Flunixin has shown to significantly improve the pain, edema 

and temperature (signs of local inflammation) in endotoxin-induced 

mastitis of dairy cattle. Flunixin treatment (1 mg/kg) was also 

associated with reduction in depression in cows with endotoxininduced 

mastitis. In another study, flunixin treatment (2.2 mg/kg) 

with an intramammary antibiotic in dairy cows with mastitis reduced 

the sensitivity to pain. In registration studies in the USA for a 

mastitis claim, flunixin (Banamine ® ) showed a significant reduction 

in pain (p=0.0199), swelling (p=0.017) and pyrexia (p

cattle ocular squamous cell carcinoma also react with equine sarcoid 

and bovine papilloma cell, implying that they may have a common 

immuno - stimulating therapy on "restricted neoplasm cell" grounds. 

Whether or not cancer cells induce immunity it is probably 

determined in part by whether they also cause inflammation. Hence 

if a metastasizing tumor doesn't invade lymphoid organs it may be 

ignored by immunity system. Conversely, on a basis of cancer cells 

possess antigen embedded on cell wall, in a cow suffering from 

cancer eye treated by other's cancer eye cells as antigens the 

immunity system destroys its own tumor cells as cell mediated 

response. Therefore the immunity system may not make any 

difference between them, as it occurs in allograft, and yet when a 

flogosis starts at the same time or later the treatment makes hasten 

curative responses. 

399 Artificial Induction of Lactation as a Remedy for Infertility 

and Stray Cow Menace 

S. Kumari, R. Paul, M. Sharma, S. Vaid 

Veterinary Polyclinic Bhuntar at Mohal, Department of Animal 

Husbandry Himachal Pradesh, Kullu, India 

In India stray cow menace is increasing day by day. Farmers do not 

want to keep the cows that become repeat breeders and cease giving 

milk. As cow slaughter and their use for meat purpose is not an 

option in India, these cows finally are left stray on the roads. These 

stray animals again pose many kind of risks to the people in the form 

of accidents, zoonoses etc. To reduce this problem 30 farmers were 

motivated to either adopt such stray cows from the roads or not to 

leave the animals stray if they had animal with such problem.Then 

these animals were induced into artificial lactation so as to make 

them useful. The artificial induction was carried out using a 

combination of Estrogen and Progesterone injections for 9 days, 

followed by an injection of Dexamethasone for five days. The 

animals started lactating within 14-15 days of treatment. With the 

priming of reproductive tract with these hormones, the cows that 

were without any anatomical or structural abnormalities, also 

conceived and delivered calves. In brief, it can be concluded that 

artificial induction of lactation can be used as a remedy to reduce 

stray cow menace, make infertile cows useful and to treat the repeat 

breeder cows (with structurally normal reproductive tracts) that are 

refractory to other treatments. 

400 Associations between Lung Lesions and Grade and Estimated 

Daily Live Weight Gain in Bull Beef at Slaughter 

P. Williams 1 , L. Green 2 

1 

Schering Plough Animal Health, Livestock Business Unit, Uxbridge, 


2 

University of Warwick, Biological Sciences, Warwick, United 

Kingdom 

A total of 645 bull beef from 15 units were examined at slaughter 

and their lungs scored for presence and severity of consolidation and 

pleurisy. Data on dead weight and carcase grade were also collected. 

Pleurisy and consolidation were correlated within cattle and both 

were associated with reduced estimated daily live weight gain 

(EDLWG) once producer variability had been accounted for in a 

hierarchical model. Consolidation had a stronger statistical 

association with EDLWG and cattle with 3, 4, 5 or 6 consolidated 

lobes had a significant reduction in EDLWG of 72, 113, 123 and 202 

grams respectively. Cattle had a maximum of four lobes with >50% 

of each lobe consolidated and those with 3 or 4 lobes with >50% 

affected had a reduced EDLWG or 141 and 194 grams respectively. 

Carcases with a higher grade were associated with a significantly 

faster EDLWG and had fewer consolidated lobes per carcase. We 

suggest that, if these associations are causal, these results highlight 

the potential cost of sub-clinical lung damage on both growth and 

carcase quality. These results may usefully be given to farmers to 

encourage changes in management to reduce lung damage in live 

cattle. 

Key words: abattoir data, cattle, lung consolidation, live weight gain, 

carcase quality 

401 Relevance of Sulphamethoxazole/Trimethoprim Susceptibility 

Ratio (19:1 or 5:1) for Bovine Mastitis Staphylococci Field 

Isolates in vitro Testing 


C. Vilela 1 , S. Nunes 1 , C. Carneiro 1 , M. Dargent-Figueiredo 2 , 

L. Cavaco 1 , M. Oliveira 1 , R. Bexiga 1 

1 Faculdade de Medicina Veterinaria de Lisboa, Centro de 

Investigaçio Interdisciplinar em Sanidade Animal (CIISA), Lisboa, 

Portugal 

2 Bayer Health Care, ANIMAL HEALTH, Lisboa, Portugal 

Antimicrobial susceptibility testing, used to identify resistance 

patterns for different bovine mastitis pathogens, is essential to issue 

recommendations for treatment at an individual or herd level. 

Sulphamethoxazole/trimethoprim (SXT) has been used for mastitis 

therapy in parenteral or intra-mammary form. This combination 

could constitute an alternative for the treatment of methicillinresistant 

S. aureus and methicillin-resistant coagulase negative 

staphylococci. Current guidelines for susceptibility testing of SXT 

issued by CLSI, using the disk diffusion method or the 

microdillution test, recommend testing a 19:1 ratio of the 2 

antimicrobials. This is based on the 19:1 human plasma 

concentrations attained when a 5:1 ratio of these antimicrobials is 

administered parenterally. However, when intramammary 

administration is performed, a 5:1 ratio is attained in the udder. We 

aimed at evaluating the minimum inhibitory concentrations of SXT 

tested at the CLSI recommended 19:1 and at 5:1.Thirty S. aureus and 

31 S. epidermidis field isolates from bovine intrammamary 

subclinical infections were used in this study. MIC values of SXT 

were determined by broth microdillution, following the CLSI (2002) 

guidelines for veterinary susceptibility testing, with a modification 

of the drug ratio for 5:1. Reference strain S. aureus ATCC 29213 

was used. MIC was considered the lowest dilution inhibiting visible 

bacterial growth. MIC values for a SXT ratio of 5:1 were compared 

to the results with the ratio of 19:1.All the S. aureus isolates tested 

were susceptible to SXT at a ratio of 19:1, while 9 of the 31 S. 

epidermidis isolates under study were not susceptible to this 

association. When testing the 5:1 ratio, 28 S. aureus and 29 S. 

epidermidis isolates showed a lower MIC value. Interpretation 

criteria for the 5:1 ratio are not available, therefore we cannot infer 

on the susceptibility. The observed difference in MIC values 

suggests that susceptibility testing results will be influenced by the 

SXT ratio used in the in vitro test. Regarding intramammary 

pathogens, a 19:1 ratio is adequate when parenteral administration is 

concerned. However, for the same pathogens, a 5:1 ratio may likely 

to reflect the drug concentration when the intramammary route is 

used. Therefore, interpretation criteria for the 5:1 ratio are required, 

in order to allow for susceptibility testing under these circumstances. 

Key words: SXT ratio, bovine mastitis, staphylococci, MIC 

402 Comparing the Effect of an Application of a Long-acting Dry 

Cow Antibiotic for Drying off with the Combined Application 

of a Long-acting Dry Cow Antibiotic and an Internal Teat 

Seal on Udder Health of Dairy Cows State up to 100 days 

Postcalving 

K. Mütze 1 , W. Wolter 2 , K. Failing 3 , B. Kloppert 2 , M. Zschöck 2 , 

H. Bernhardt 1 

1 Institut für Landtechnik Justus Liebig Universität Giessen, Giessen, 

Germany 

2 Landesbetrieb Hessiches Landeslabor, Abteilung Veterinärmedizin, 

Giessen, Germany 

3 Arbeitsgruppe Biomathematik und Datenverarbeitung des 

Fachbereichs Veterinärmedizin der Justus-Liebig-Universität 

Giessen, Giessen, Germany 

The objectives of this study were to clarify whether the application 

of an internal teat seal (OrbeSeal ® , Pfizer/ Karlsruhe) in 

combination with a long-acting intramammary dry cow antibiotic 

offers an additional protection against new intramammary 

infections (IMI) during the dry period and has influence on IMI 

status, clinical mastitis events and number of somatic cells up to 

100 days postcalving compared with a single application of a longacting 

intramammary dry cow antibiotic. A field study was carried 

out from July 2006 until March 2007 with 152 cows of 12 Hessian 

dairy farms. Before the final milking on the day of dry off duplicate 

quarter milk samples have been taken for cytobacteriologic 

analysis. The cows of each farm were parted into 2 groups by 

chance. After milking all 4 quarters of the cows of group AB were 

infused with long-acting dry cow antibiotic. Cows of the second 

group AB+OS additionally to the long-acting antibiotic the internal

teat seal OrbeSeal ® was applied. For reasons of statistic evaluation 

we chose pairs of one treated quarter (AB+OS) and one control 

quarter (AB) each, which had been similar according to their 

previous state (cell content, bacteriologic state) before dry off. On 

the whole we evaluated data of 136 cows. The results of the 

California Mastitis Test between 1 and 5 days postcalving were 

better for group AB+OS (57 positive reactions) than for group AB 

(85 positive findings) (p=0,0004).During the period 6-14 days, as 

well as 35-56 days after calving quarter milk samples were 

examined cytobacteriologically. During both terms we proved 

numerically tending less pathogens in group AB+OS than in group 

AB. There have been statistically significant differences in the 

proof of Corynebacteria spp. with p=0,0471 during the term 6-14 

days as well as with p=0,0161 during the term 35-56 days and 

concerned the aesculine positive streptococci (S. uberis) during the 

term 35-56 days postcalving (p=0,0161). Clinical mastitis events 

occurred in 15 quarters of the AB+OS group and in 25 quarters of 

the AB group between calving and 100 th day of lactation 

(p=0,0257). In comparison with the in dairy farms commonly used 

dry cow antibiotics we could reduce in the frame of this study by the 

additional application of a teat seal to the long-acting antibiotic the 

part of quarters infected by environmental mastitis pathogens 

(mainly S. uberis) as well as the rate of clinical mastitis events 

during early lactation. 

Key words: bovine mastitis, dry cow therapie, teat sealer, udder health 

403 Effect of Cefalonium on Intramammary Infections and on 

Milk Production of Heifers before their First Calving 

L. Jimenez 1 , R. Timon 1 , J. Marco 2 

1 

Servet Talavera, Dairy Milk Quality Centre, Talavera de la Reina, 

Spain 

2 

Public Health Laboratory, Health Department, País Vasco, Spain 

Objectives: To demonstrate the efficacy of Cepravin V.S. in heifers 

studying: 

Prevalence of intramammary infections (IMI) 

– Milk production 

– Somatic cell count (SCC) 

Materials and Methods: The randomized study included 158 

Holstein heifers: 78 as a control group (CON) and 80 in the treatment 

group (CEP).Samples were collected from each quarter, according to 

National Mastitis Council (NMC), 60 days before calving (BC 

sample), on day 7 post calving (d7 pc sample), and on day 21 post 

calving (d21 pc sample). On days 50 and 100 (d50 and d100 

pc sample, respectively) a pooled sample was collected. 

Microbiological testing was performed according to NMC.Heifers 

from the CEP group received Cepravin ® V.S. (cefalonium, 250mg) 

after collecting the BC sample.Milk production and SCC was 

assessed on a monthly basis. 

Results: The prevalence of IMI per group was not significant /Table 1). 

Table 1. IMI prevalence (%) 

Group/ BC d7d pc d21d pc d50 pc d100 pc 

Time (%) (%) (%) (%) (%) 

CEP 75,6 52,1 55,7 42 47 

CON 65 56,6 57,3 40 51,5 

N 158 149 145 138 140 

The cure rate on day 7 was higher in the CEP group (p0,01), with lower arithmetic means for the CEP group (min-max: 

100.000 -224.000 vs. 146.000 -348.000 cells/ml respectively for CEP 

and CON). 

Table 3. Daily milk production (l; lactation standardized at 305 days) 

Month 1 2 3 4 5 6 7 8 9 10 

CEP 30,9 36,2 38,6 37,9 36,5 35,1 32,0 30,4 27,6 26,4 

CON 30,2 34,4 35,0 34,5 33,6 31,5 28,7 25,9 24,3 22,8 

Concerning dairy performance, the CEP group produced 911 litres 

more than the CON group (10.057 vs. 9.146 l; p= 0,0015; Table 3). 

Conclusions: Cepravin shows a preventive and curative effect for IMI 

caused by CNNSS. A 911 l (10.057 vs. 9.146 l; p= 0,0015) increase of 

milk production of treated heifers was found. These results support the 

advisability and profitability of systematic treatment of dairy heifers 60 

days before calving with Cepravin VS. 

Key words: cefalonium, heifer, milk production, CNS 

404 Widespread Distribution of pS194-mediated Streptomycin 

Resistance in Staphylococci of Farm Animal Origin 

J. Bjorland 1 , T. Mork 2 , S. Waage 3 , M. Sunde 2 

1 

Norwegian School of Veterinary Science, Production Animal 

Clinical Sciences, Sandnes, Norway 

2 

National Veterinary Institute, Department of Animal Health, Oslo, 

Norway 

3 

Norwegian School of Veterinary Science, Production Animal 

Clinical Sciences, Oslo, Norway 

Plasmids carrying antibiotic resistance determinants are considered to 

be key elements in the evolution of antibiotic resistance among 

bacteria. The aminoglycoside streptomycin is commonly used in 

combination with penicillin to treat mastitis in farm animals. The 

objective of the study was to elucidate the role of plasmids in 

connection with streptomycin resistance in staphylococci. During 

1997-1998, 99 isolates of streptomycin-resistant Staphylococcus 

aureus and 23 isolates of coagulase-negative staphylococci (CoNS) 

were collected. All CoNS isolates and 89 S. aureus isolates originated 

from mastitic milk from dairy cows in different areas of Norway, three 

S. aureus isolates were from two dairy goats and one sow with mastitis, 

and two S. aureus isolates were from food products based on 

unpasteurized milk. Included were also two S. aureus isolates from 

Sweden and three from Finland, all being associated with bovine 

mastitis. StaphZym was used for species determination of CoNS. 

Genotyping of S. aureus isolates was carried out using pulsed-field gel 

electrophoresis (PFGE). Eighty-two S. aureus isolates (82.9 %), 

including one Swedish, two Finnish and those from food, and 20 CoNS 

isolates (86.9 %) contained a plasmid with a particular endonuclease 

restriction profile. The sequence of the plasmid (4399 bp) is 99% 

identical to the previously reported pS194 (4397 bp). It consists of four 

open reading frames including the aadE gene encoding an adenylyltransferase 

required for streptomycin resistance. The CoNS species 

were identified as S. chromogenes (6 isolates), S. simulans (6 isolates), 

S. epidermidis (4 isolates), S. sciuri (1 isolate), and Staphylococcus sp. 

(3 isolates). PFGE typing of 58 randomly selected pS194-containing S. 

aureus isolates, representing 44 dairy herds, revealed 39 different 

PFGE types. S. aureus isolates of indistinguishable PFGE types were 

found in the sow and two dairy cows, and in a dairy cow and a goat. In 

conclusion, streptomycin resistance in staphylococci causing mastitis 

in dairy herds in Norway is to a large extent provided by the plasmid 

pS194, which is present in several S. aureus genotypes and CoNS 

species. Certain pS194-containing S. aureus strains are able to infect 

both different domestic ruminants and sows. 

405 Pharmacokinetics and PK/PD Integration of Danofloxacin in 

Lactating Dairy Cows with Subclinical Mastitis caused by 

Staphylococcus aureus 

M. Lucas, L. Marchetti, A. Lambertini, J. Errecalde, N. Mestorino 

Faculty of Veterinary, UNLP, Pharmacology, La Plata, Argentina 

The objective of this study was to investigate danofloxacin (DNF) 

pharmacokinetic (PK) profile in lactating dairy cows with 

subclinical S. aureus mastitis, and calculate the corresponding 

Miscellaneous 295

pharmacokinetic/pharmacodynamic (PK/PD) parameters. Six 

Holstein lactating cows with subclinical mastitis in at least one 

mammary quarter were selected. DNF 18% (Advocin ® ,Pfizer Inc, 

Argentina) was administered subcutaneously (10 mg/kg single 

dose). Blood and quarter milk samples were collected at different 

times until 96h after drug administration. Milk and plasma DNF 

concentrations were measured by HPLC with fluorescence 

detection.The MIC 90 was determined in 51 S. aureus strains to 

calculate PK/PD parameters. PK parameters were determined by 

non-compartmental analysis. Each mammary quarter was considered 

independently (n=24), since our purpose was to compare the 

findings in mastitic quarters (n=8) against healthy ones (n=16) and 

in quarters of high-producing (HP) cows (n=12) against lowproducing 

(LP) cows (n=12). The PK/PD parameters calculated were 

AUC 0-24h/MIC 90 (area under the concentration vs time until 

24h/MIC for 90% of the isolates), AUC 0-∞/MIC 90 (AUC until 

infinity/MIC 90) and T>MIC 90 (time during which the concentrations 

exceed the MIC 90).DNF penetration into milk was fast. DNF 

concentrations determined in milk (C max=2.7±0.8 µg/mL) were 

higher than those found in plasma (C max=1.4±0.2 µg/mL) and milk 

bioavailability (F milk) was 4.2.C max in plasma occurred 1.2h postadministration, 

while C max in milk occurred 6h post-administration. 

T1/2 in milk (8.5h) was longer than T1/2 in plasma (4.4h). The PK 

parameters shows that milk concentrations decrease slower than 

plasma ones. According to the level of production, DNF was 

eliminated slower from quarters of LP cows. Concomitantly, the 

mean residence time (MRT) in milk from quarters of LP cows was 

longer than the MRT obtained in milk from quarters of HP cows. The 

C max and the C max/MIC 90 resulted larger in milk from quarters of LP 

cows. According to health status, it was observed that the MRT was 

longer in mastitic quarters. The level of production and the quarter 

health status could modify the PK and PK/PD parameters of DNF in 

milk of lactating cows treated parenterally. Consequently, we 

consider that therapeutic protocols should be adjusted according to 

susceptibility and the MIC of the bacteria involved and the level of 

production of the animal to be treated. 

Key words: danofloxacin, pharmacokinetic/pharmacodynamic, milk, 

mastitis, cows 

406 Epidemiology of Staphyloccoccus aureus Intramammary 

Infection in Dairy Cows in Slovenia 

O. Podpecan 1 , P. Zrimsek 2 , M. Pate 3 , M. Ocepek 3 , I. Zdovc 3 , 

A. Pengov 3 

1 

Savinian Veterinary Policlinic, Ruminants, alec, Slovenia 

2 

Veterinary faculty Ljubljana, Clinic for Reproduction and Horses, 

Ljubljana, Slovenia 

3 

Veterinary faculty Ljubljana, Institute for Microbiology and 

Parasitology, Ljubljana, Slovenia 

The main objective of the study was to determine factors associated 

with epidemiology of S. aureus intrammamary infections in dairy 

cows. A total of 763 dairy cows and 2978 udder quarters from 42 

herds were included in the study. Lactation number, body condition 

score and somatic cell counts (SCC) were calculated. Bacteriological 

determination of isolates and genotyping of S. aureus were 

performed using the Api-Staph test and pulsed-field gel 

electrophoresis (PFGE). Antibiotic resistance rates of S. aureus 

isolates were determined using the agar disk diffusion test. 

Statistical comparison of the results was performed using Kruskal- 

Wallis One Way Analysis of Variance on Ranks, Dunn´s test and 

Mann-Whitney-U test. The prevalence of S. aureus intramammary 

infection on a herd level was 33.7%. SCC in milk from udder 

quarters infected with S. aureus were significantly higher, 

irrespective of the lactation number, comparing with bacteriological 

negative udder quarters (P

Key words: semen kinematics, semen, fertility, CASA 

409 Effect of Antioxidant Addition on in vitro Semen Quality and 

in vivo Fertility of Frozen-Thawed Bovine Semen 

M. Griga 1 , H. Nehring 2 , C. Leiding 1 

1 Artificial Insemination Center BVN, Animal Reproduction and A.I, 

Neustadt Aisch, Germany 

2 Institut für Fortpflanzung landwirtschaftlicher Nutztiere, Animal 

Reproduction and A.I, Berlin, Germany 

Reactive oxygen species (ROS) play a significant role in many of the 

sperm physiological processes and they also trigger many 

pathological processes. Favourable effects of antioxidant molecules 

on sperm motility have been reported in the literature. The objective 

of this study was to examine the influence of antioxidant molecules, 

content in to the Oxyfree ® Premix (IMV, Technologies, France), on 

the motility and fertility when added to bulls semen diluted with a 

egg yolk extender (Triladyl ® , Minitueb, Germany). Four ejaculate 

from each six Fleckvieh bulls were split into four fractions. Two 

fractions were diluted to a final concentration of 14 x 106 

sperm/straw and another two fracton of 8 x 10 6 sperm/straw. One 

fraction with 14 and one with 8 x10 6 sperm/straw were diluted with 

extender containing Oxyfree. Frozen-thawed semen were evaluated 

for motility by a CASA system (SpermVision ® , Minitueb, Germany) 

30 and 120 minutes after thawed and acrosomal integrity and 

spermatic membrane integrity by staining with fluorescein 

isothiocyanate-conjugated Arachis hypogea agglutinin (FITC-PNA) 

in combination with propidium iodide and analysed by flow 

cytometer (Beckman & Coulter, USA). ROS was determined by 

flow cytometer by staining with BODIPY 581/591 , 

dihydrorhodamine 123 (DHR) and dichlorofluorescin diacetate 

(DCFH), 15 minutes, 2 hours and 4 hours post-thaw and also 15 

minutes with oxidative stress by adding of butyl hydroxide and 

hydrogen peroxide. Post-thawed semen evaluation schowed that 

motility (mean ± SD) was increased (p

Bovine Immunodeficiency Virus (BIV) is a Lentivirus, which can 

cause varieties of infectious complications due to immune system 

dysfunctions and induce remarkable economic losses to dairy cattle 

industry. Considering the probability that BIV infection can be 

worldwide distributed and giving attention to the fact that till now 

not only any epidemiological survey but also any case study have not 

been done, therefore this is the first investigation about BIV 

infection in Iran. In this study 197 Holstein cattle slaughtered in 

Meysam abattoir in Tehran province were surveyed. The 

investigation began from the October 2005 and ended in March 

2006. The purpose of the survey was to determine the infection rate 

of BIV and the effects of the infection on the clinical and 

pathological parameters. Nested- PCR was used in the diagnosis of 

the infection.The results of the study showed that total infection rate 

was 16.2% (32/197). The animals with four pair’s permanent teeth 

had the highest (26.8%) infection rate (0.01

Objective of study: Currently it is assumed, that the lung of newborn 

calves expands and starts gas exchange quite shortly after birth. Up to 

now, it was not shown at what time during the first weeks of life, 

different lung areas of bovine neonates show a maximum gas exchange 

rate. Therefore the lung development of bovine neonates within the first 

three weeks post natal was examined using CT. 

Material and methods: For the ongoing examinations of the first three 

weeks post natum 15 healthy Holstein Frisian calves, born without any 

complications, were chosen. Their vitality was checked with the 

APGAR-Score (>7.5) and blood-gas-analysis.The lung function was 

measured segmental in a spiral CT -PQ 2000- of the brand Picker 

International. In order to examine the thorax area, the animals were 

slightly sedated (Xylazinhydrochloride, Impfstoffwerke Dessau, 0.09 

mg/kg body weight). The Region of interest (ROI) was chosen in a way 

that lung tissue without larger supply-structures (such as vessels) was 

measured (Roi sqnn 12): dorsal left and right quadrant, ventral left and 

right quadrant; from cranial to caudal direction. The CT-measuring 

times were: 0.5-1 h, 6 h, 12 h p. n., 1 st , 2 nd and 3 rd week p. n.The 

statistical analysis was made with SPSS 14.0 using variance analysis 

and Pearson’s correlation coefficient. 

Results: 1. Immediately p. n. the two dorsal lung areas are ventilated, 

with a delay of about 12 hours also the ventral ones (p < 0.001). 2.The 

expansion of the lung-alveoli increases from cranial to caudal. The time 

until the cranial lung areas are completely ventilated was two weeks, 

for the caudal areas three weeks. 3. In the cranial area, the left half of 

the lung is better ventilated than the right. (1 st hour p. n. until 2 nd week 

post natum p < 0.02 to 0.001) whereas the right caudal lung area 

participates more intensively after the 12 th hour post natum until the 

third week post natum in the increasing gas exchange than the left area 

(p < 0.05 to 0.001). 

Conclusions: A complete gas exchange activity within the bovine lung 

is only given with the beginning of the second week post natum. It was 

shown that there are significant differences in the development of the 

stability of lung alveoli comparing cranial and caudal lung segments as 

well as dorsal and ventral lung areas. 

Key words: neonatal calves, lung ventilation, lung computer 

tomography 

440 Effects of Meloxicam Therapy on the Behaviour and Welfare 

of Dairy Calves with Neonatal Calf Diarrhea Complex 

C. Todd 1 , S. Millman 1,2 , T. Duffield 1 , D. McKnight 3 , K. Leslie 1 

1 University of Guelph, Population Medicine, Guelph, Canada 

2 Iowa State University, Population Medicine, Ames, United States 

3 University of Guelph, Kemptville Campus, Kemptville, Canada 

Infectious disease poses a major welfare challenge for neonatal dairy 

calves. Opportunities for supportive therapy which may alleviate 

animal suffering during disease states need to be identified and 

formally evaluated. The objective of this study was to examine 

whether meloxicam (Metacam ® 20 mg/mL solution for injection for 

cattle, pigs and horses, Boehringer Ingelheim) is an effective 

supportive therapy for the alleviation of sickness and pain-related 

behaviours in cases of neonatal calf diarrhea complex. For this 

double-blind controlled trial, 62 Holstein bull calves were purchased 

at birth and transported to a calf research facility. At the naturally 

occurring onset of diarrhea, the calves were enrolled in the study and 

randomly assigned to receive a single subcutaneous injection of 

meloxicam (0.5 mg/kg BW) or an equal volume of placebo solution. 

Starter ration and milk intakes were recorded daily for each calf until 

56 days of age. Following onset of diarrhea, calf feeding behaviour, 

lying posture, standing posture and activity were evaluated over a 

five day observation period. During the study, 56 calves presented 

with clinical signs of diarrhea and were subsequently treated with 

meloxicam (n=28) or placebo (n=28). Meloxicam-treated calves 

began consuming starter ration significantly earlier (p1/50 (Pomona 11/24 vs. 23/23, D 54% CI 28-80%, P

significantly lower in groups vaccinated at 1 month (P

University of Tehran, Animal Science Department, Karaj, Tehran, 

Iran 

We evaluated the effects of Monesin and propylene glycol on blood 

metabolites, rumen NH 3 and rumen pH of 16 primiparous and 

multiparous Holstein lactating cows (60-30 DIM, Milk production 

33±3 Kg/day) between September and November of 2007. Cows 

were used in a completely randomized design with 4 treatments 

(additives) and 4 replicates (cows) and were fed a balanced total 

mixed ration (60% concentrate and 40% forages) with or without 

additives. Cows in group I were fed TMR without additive (control), 

Cows in group II were fed TMR with 335 mg/day monensin, Cows 

in group III were fed TMR with 400 ml/day propylene glycol and 

Cows in group IV were fed TMR with 335 mg/day Monensin and 

400 ml/day propylene glycol. Data were analyzed using a mixed 

model for repeated measurement. Cows treated with Monensin 

and/or propylene glycol had significantly more concentrations of 

plasma glucose, (51.92, 56.15, 56.92 and 58.56 mg/dl for groups I, 

II, III and IV respectively; P

5.00+0.67 and 4.94+0.15 mEq/l in the blood plasma of yaks in the 

three trimesters of pregnancy, respectively. 

1062 Relationship between Hepatotoxic Compounds and 

Photosensitization in Cattle 

J. Seixas 1 , P. Peixoto 2 , C. Pinto 3 

1 

Universidade Federal Rural do Rio de Janeiro, Curso de Pos 

Graduacao em Ciencias Veterinarias, Seropédica, Rio de Janeiro, 

Brazil 

2 

Universidade Federal Rural do Rio de Janeiro, Instituto de 

Zootecnia, Seropédica, Rio de Janeiro, Brazil 

3 

Serviço de Desenvolvimento Agrário de Sao Miguel, Ponta Delgada, 

Portugal 

Most photosensitization in cattle is caused by phylloerythrin 

retention consequential to liver damage caused by hepatotoxic 

compounds. Injury to hepatic canalicular membranes and subsequent 

cholestasis occurs in Lantana spp poisoning, although there is also 

significant parenchimal lesions. Primary hepatocellular injury is 

caused by the consumption of Myoporum laetum. Photosensitization 

caused by sporidesmin from Pithomyces chartarum is characterized 

mainly by marked fibrosis biliary obstruction may be caused by the 

presence of birefringents crystals. The consumption of plants 

containing saponins causes icterus which has, as one of the main 

characteristics, crystal-associated cholangiopathy. Poisoning by 

saponins is developed after consumption of Tribulus terrestris, 

Panicum spp, Narthecium ossifragum and Brachiaria spp. This work 

describes the main hepatotoxins causing cattle photosensitization 

and relates them to their pathogenetic effects. The pentacyclic 

triterpenoids (present in Lantana spp) cause intrahepatic cholestasis 

with the inhibition of bile secretion. The metabolism of essential oils 

of furanosesquiterpenes (M. laetum) cause hepatic necrosis. Saponin 

poisoning normally occurs with hepatic biliary crystal obstruction. 

Injury to the biliary ducts caused by the sporidesmin is characterized 

by severe pericholangitis caused by the generation of oxygen free 

radicals and resulting in fibrosis. There is no scientific consensus 

about T. terrestris and N. ossifragum poisoning concerning hepatic 

target lesions: hepatocellular or biliary system. Brachiaria spp 

poisoning should be better included in the group of primary liver 

parenchymal damaging plants, because of although it occur, in a 

small percentage of cases, deposition of crystals birefringents in the 

biliary ducts, those lesions are, in general, enough located. On the 

other hand, ultramicroscopy reveals marked deposition of crystals 

inside hepatocytes, with development of degenerative alterations. It 

is also very characteristic the formation of numerous foam cells, 

general under form of foci (crystals and remainders of necrotic 

hepatocytes inside the macrophages) (Lemos et al., 1997; Driemeier 

et al., 1998; Gomar et al., 2005; Seixas & Peixoto, 2007). Those 

characteristics indicate that the phylloerythrin retention in case of 

Brachiaria spp poisoning is doing in the hepatic parenchyma and not 

in the biliary system. 

Key words: photosensitization, hepatotoxic compounds, saponins, 

sporidesmin, Brachiaria spp 

1063 Urinary Bladder Rupture in Consequence of Urolithiasis in 

Nelore Cattle 

R. Godoy, L. Gouvea, C. Leite, C. Pereira, FH. Ximenes, 

L. Gontijo, R. Ferreira II, JR. Borges 


Objective: The case report is of Nelore cattle, 28 months, male, 

presenting obstructive urolithiasis and urinary bladder rupture, 

examined at the School Hospital of Large Animals at the University of 

Brasília. 

Material and method the animal had anuria, tachycardia, 

dehydration, abdominal pain and azotemia. It was on exhibition for 

24 days, being fed with hay, silage and concentrate. Trans-rectal 

palpation showed a filled bladder, soon after that the animal expelled 

a calculi. The urine had reddish colour, density 1,010, pH 7.0, 

protein (++), haemoglobin (++++) and macroscopically a lot of 

smaller stones could be visualized. The treatment during 

hospitalization was of continuous intravascular fluid, vitamin C 

(10mg/Kg - TID - 3 days), enrofloxacin (5mg/Kg - SID - 5 days), 

flunixim meglumine (1.1 mg / kg - SID - 2 days), oral-ruminal 

hydration and restricted diet of grass. The improvement was evident 


by the obtention of normal colored urine in the end of the second day 

of treatment. The animal was released with guidance of restriction of 

continuous concentrate feed as treatment. Forty days after, it 

returned to the Veterinary Hospital showing symptoms worsened to 

his previous hospitalization, again with a history of beind fed with 

concentrate. The animal was apathetic with dirty nostrils, reluctant 

to move, tachycardiac, with severe dehydration, tachypnea, 

abdominal pain and ruminal. Paracentesis revealed large quantities 

of fluid with odor resembling urine. The presence of a greater 

concentration of creatinine in the peritoneal fluid (42.5 mg / dL) 

compared to its serum concentration (10.5 mg / dL) proved the fact 

of it being urine. Exploratory laparotomy was performed and 

confirmed the rupture of bladder. 

Results: after analysis of laboratory tests, clinical evaluation and 

laparotomy, euthanasia was performed. At necropsy, macroscopically 

confirmed the bladder rupture and found to be calculi of 2.5 x1, 0cm the 

cause of obstruction in urethra near the glans. 

Conclusion: It was concluded that treatment done during the first 

hospitalization was effective, however, animals with a predisposition 

to the formation of calculi that continue to receive food rich in 

concentrate and are subjected to constant stress, may have recurrent 

obstructions. Comparison of the levels of creatinine, in the serum and 

peritoneum, proved to be an important tool in the diagnosis of the 

presence of urine in the abdominal cavity. 

Key words: calculi, urethra obstruction, creatinine 

1064 Carcass pH and Temperature of Supply Animals with 

Organic Chromium, of Nellore and F1 Brangus x Nellore 

Cattle on Pasture System 

A. Jorge 1 , A. Polizel Neto 1 , P. Moreira 2 , A. Ramos 1 , J. Souza 3 , 

R. Pinheiro 1 , C. Francisco 1 

1 UNESP- School of Veterinary Medicine and Animal Sciences, 

Animal production, Botucatu, Brazil 

2 UFMT, Animal Production, Sinop, Brazil 

3 UFPR, Animal Science Department, Palotina, Brazil 

Chromium works as component integral and biologically that 

potential the insulin action in the cells, stimulating there of keep the 

glycogenic and this affecting post mortem pH. The aim of this study 

was evaluate the effect of the organic chromium supplementation in 

the carcass pH curve and freezing, of the two racial groups of steers, 

Nellore e F1 Brangus x Nellore, on pasture system. Were used two 

groups composed 18 animals each (9 Nellore e 9 F 1 Brangus x 

Nellore), submitted to two experimental treatments, with and 

without organic chromium supplementation. The animals were 

distributed in the treatments at 210 days until at 600 days of age, 

when the animals were slaughed, and gauged the pH and the 

temperature at 2, 10 and 24 hours after animals died. Not was 

detected racial group, treatment and interaction influence to any 

characteristics evaluated. The conclusion was that the organic 

chromium supplementation to beef cattle, on pasture system, not 

influences the carcass pH curve and temperature. 

Key words: bovine, chromium, meat, minerals, supplementation 

1065 Evaluation of Carcass Alteration, by Ultrasound Measures, 

of Nellore and F1 Brangus x Nellore Steers Supply with 

Organic Chromium in Pasture System 



1 UNESP School of Veterinary Medicine and Animal Sciences, Animal 

Science Department, Botucatu, Brazil 

2 UFMT, Animal Science Department, Sinop, Brazil 


As chromium is related to insulin, supplementing with bio available 

organic molecules containing chromium could be quite important for 

improving muscular growth and carcass quality. Thus the objective of 

this experiment was to evaluate the carcass alterations, by ultrasound 

measures of the Nellore and F 1 Brangus x Nellore steers supply with 

organic chromium, in pasture system. Were used two groups with 18 

animals each (9 Nellore and 9 F 1 Brangus x Nellore), submitted to two 

experimental treatments: with and without organic chromium 

supplementation. Were measures, in two experimental moments, at 450 

(I) and 600 (II) days of age, the rib eye area (REA_I and REA_II), in 

cm 2 ; the sub cutaneus fat thickness at the back and at the rump, in mm,

y ultrasound measures; and the gains, calculated by difference 

between I and II, of the rib eye area (GREA), of the subcutaneous fat 

thickness at the back and at the rump; and took the lives weight, in kg. 

No differences were detected between racial groups for any variable, 

nor was any treatment x breed interaction detected; however was 

observed difference only REA_II and GREA, between treatment. 

Conclusion that organic chromium supplementation improved rib eye 

area gain, and did not affect the subcutaneous fat deposition. 

Key words: bovine, carcass, growth, rib eye area, subcutaneous fat 

1066 Different Supplementation Frequency of the Steer in 

Pasture, on Rain Season: Ruminal Parameters 



1 UNESP School of Veterinary Medicine and Animal Sciences, Animal 

Science Department, Botucatu, Brazil 

2 UFMT, Animal Science Department, Sinop, Brazil 


The aim of this study was to determine ruminal pH and quantify ruminal 

ammonia concentration in different frequency supplementation of the 

steer in good pasture, on rain season. Were used four Zebu animals, 

castrated, and rumen fistuladed, distributed in latin square 4x4, 

composed by four frequency and four experimental period. The 

supplements were offered in quantity equivalent 0.2 % body weight per 

day (1.4 % BW/week), in different weekly frequency, at 10:00: control 

(only mineral supplement), two times a week, three times a week and 

daily. Were collected 50 mL of the ruminal liquid and determined the 

pH and quantified the ammonia concentration in three times after 

supply, at 0,4 e 24 hours. Not was observed frequency influence on 

ruminal pH, and note minor value four hours after supply among the 

times studied. About the ammonia concentration among the times was 

observed greater value four hours after supply; and among the 

treatments was found greater values to supply animals, independently of 

the frequency. The different supplementation frequency on pasture not 

influenced the ruminal pH and ammonia concentration. 

Key words: ammonia, bovine, pasture, rumen, ruminal pH 

1067 Increased Oxidative Stress in Liver Biopsies from Cattle 

with Hepatic Dysfunction 

M. Abd Ellah 1 , K. Okada 2 , M. Goryo 3 , J. Yasuda 2 

1 


Assiut, Egypt 

2 

Iwate University, Animal Medicine, Morioka, Japan 

3 

Iwate University, Vetrinary Pathology, Morioka, Japan 

The goal of the present study was to measure hepatic antioxidants 

enzyme activities in bovine liver biopsies from cows with 

experimentally induced hepatic dysfunction. Seven cows were 

subjected to study; five cows were injected intraperitoneally with 

DL-ethionine (12.5 mg/kg B.W.) and the remaining two cows were 

injected with normal saline (0.9% NaCl) and kept as control. 

Ultrasonography guided liver biopsy specimens were collected at 0, 

4, 7 and 10 days after injection. Liver biopsies were washed twice in 

cold 0.9% NaCl, and then homogenized in 20 volumes (per weight 

tissues) of cold buffer (50mM TRIS-HCL, PH 7.5) and the 

antioxidants parameters were measured in the supernatant fluid. No 

histopatological changes were observed in the control group. On the 

other hand, liver of cows treated with ethionine showed different 

degrees of cell swelling and glycogen degeneration in days 4, 7 and 

10. Biochemical analysis of the hepatic tissues revealed significant 

increases in hepatic total glutathione level in days 7 (p

potable water and food, avoiding stress, Chapter 7 obligations with 

society. To compete version of the ethics code consultation you can 

go to www.ammveb.net 

Key words: bioethic 

1070 Pollution et Syndrome des Corps Étrangers 

M. Tlidjane, Z. Rouabah, N. Alloui, B. Safsaf 

Batna University, Department of Veterinary Medicine, Algeria 

This study has been archived in three areas of the wilaya of Batna. 

The first one is located near a great domestic waste discharge at open 

sky (Azab).The second area consisted of medium waste discharge 

(El-Madher) and the third one without a fixed public waste discharge 

(Sériana). Three types of investigation were performed. The first one 

concerned the degree of pollution of pasture fields. It showed that 

pollution is very high in the immediate nearby of the discharge and 

this pollution had a direct effect on the animal health. The second 

survey at slaughter-house showed that animals that had ingested the 

most non-biodegradable foreign bodies come from breedings located 

near great or average waste discharge. The survey on the frequency 

of the foreign body syndrome showed a clear relation-ship between 

the frequency and the degree of pasture field pollution. This finding 

reveals that non-protected waste discharges constitute the main 

source of pasture fields pollution and have a direct impact on animal 

health. 

1071 Genetic Relationships between Linear Type Traits, Somatic 

Cell Score and Longevity in Holstein Cows of Iran 

M. Bakhtiarizadeh, M. Moradi Shahr Babak, A. Pakdel, 

M. Motlagh Khodaie 

University of Tehran, Faculty of Agronomy and Animal Science, 

Karaj, Iran 

The objective of the present study was to estimate the genetic 

parameter for 13 linear type traits, somatic cell count and longevity 

in Holstein population of Iran.Two set data including 3000 (for SCS) 

and 12226 (for longevity) first lactation records on cows distributed 

across 219 and 1500 herd-year-season groups respectively. SAS 

software and GLM proc were used to fit the model. Genetic 

parameters were estimated by restricted maximum likelihood with 

an animal model and ASREML software. Heritability estimates for 

the type traits varied from 0.033 (foot angle) to 0.29 (stature).genetic 

correlation among type traits ranged from 0.01 (suspensory ligament 

and Body Depth) to 0.83 (stature and rump length). Genetic 

correlation among type traits and longevity (Functional Length 

Productive Life) ranged from 0.41 (udder depth) to -0.5 (chest with) 

and among type traits and SCS ranged from 0.85(rear udder width) 

to -0.6 (fore udder attachment). Most of the genetic correlations 

among type traits (except foot angle, fore udder attachment, rear 

udder height, suspensory ligament and fore teat placement) with 

longevity and SCS had a reciprocal sign or in other words cows with 

more longevity had a lower SCS and cows with lower longevity had 

a more SCS. In result more attention should pay to these traits in 

breeding programs for better animal welfare and profit.Negative 

genetic correlation among foot angle, udder depth, fore udder 

attachment and rear leg side view with SCS and also positive genetic 

correlation among suspensory ligament, udder depth, fore teat 

placement and rear leg side view with longevity indicate that these 

traits can be used within a selection index in national progenytesting 

program. In general results of this study indicated that 

genetically taller, wider, deeper with bigger rump length cows have 

a more SCS and lower longevity. 

Key words: type traits, SCS, longevity 

1072 Effect of Cadmium Ion on Peroxidase Activity of Serum in 

Cow: (An in vitro study) 

H. Tayefi-Nasrabadi 1 , Gh. Moghaddam 2 , Z. Rajabi 3 , Gh. Dehghan 4 

1 Faculty of Veterinary Medicine, University of Tabriz, Basic 


2 Faculty of Agriculture, University of Tabriz, Animal Sciences, 

Tabriz, Iran 

3 

Faculty of Veterinary Medicine, University of Tabriz, Clinical 


4 

University of Tabriz, Faculty of Veterinary Medicine, Tabriz, Iran 


Cadmium is a widely distributed metallic pollutant of our 

environment, which can be absorbed into biological systems through 

direct uptake as well as by accumulation in food chains. It has toxic 

effects on all living systems, whether human, animal, plant or 

bacterium. Peroxidases are important detoxifying enzymes system 

serving to rid cells of excess H 2O 2 under normal and stress 

conditions, including contamination by toxic levels of heavy metals. 

However, extreme stress conditions may affect the activity of the 

detoxification enzymes themselves. For clarifying some possible 

mechanism of cadmium toxicity, the effect of increasing amounts of 

Cd 2+ ion on peroxidase activity was investigated in vitro in serum of 

cow. The H 2O 2-mediated oxidation of o-dianisidine was used to 

assess the peroxidase activity. Results show that after preincubation 

of serum with 0.25-100 mM Cd 2+ concentration for 5 minutes, 

peroxidase activity was inhibited compared to the control and 

decreased rapidly with increasing metal concentrations. The enzyme 

was completely inhibited after 5 min preincubation in 100 mM Cd 2+ . 

When the preincubation of serum and Cd 2+ was prolonged to 0.5, 1 

and 24 hours, the enzymatic activity decreased more rapidly with 

increasing metal concentration, and the enzyme was completely 

inhibited at lower metal concentrations (at 50 mM Cd 2+ after 30-min 

preincubation, at 30 mM Cd 2+ after 60-min preincubation and at 5 

mM Cd 2+ after 24-h). By considering of long biological half-life of 

cadmium in body of animals, it suggested that the damage caused by 

exposure to heavy metals is often not only dose-dependent, but also 

time-dependent. Even though detoxifying enzymes may not show 

any effect after brief exposure to low concentrations of heavy 

metals, prolonged incubation will affect the enzymatic activity, 

leading eventually to complete inactivation. 

Key words: cadmium, time dependent, peroxidase activity, serum, 

cow 

1073 In situ Dry Matter Degradation Kinetics of Treated and 

Untreated Sainfoin (Onobrychis Viciifolia) 

H. Khalilvandi, K. RezaYazdi, M. Dehghan-Banadaky, 

N. Vahdani, H. Khazanehei 

University of Tehran, Animal Science Department, Karaj, Tehran, 

Iran 

Sainfoin is tanniferous legume forage with CT concentration of 2.5 

to 7.7 % of dry matter. In order to investigation of different 

chemicals to improving degradation characteristics of Sainfoin, an In 

situ Experiment carried out using 3 ruminally fistulated Holstein 

cows. Samples of forage were chapped 3-5 cm length, and 

then treated with NaoH (0.05 M) and Sodium Bicarbonate (0.1 M) 

solutions with forage to reagent volume ratio of 1:4 (W/V). 

Treatments carried out in 25 ° C temperature, for 20 min, with hand 

shaking. 5 % solution of PEG (6000 MW) was sprayed to forage 

with 1:1 ratio of 1:4 (W/V). Treated forages then expose to 40 ° C 

temperature in a forced air oven, for 48 hour. All forage samples 

grind to pass 2 mm screen size (Wiley mill). 5 g of forage samples 

were weighed into nylon bags (10_20 cm) with 53 µm pore size. 

Duplicates were incubating for 4, 8, 12, 24, 48, 72 and 96h in ventral 

rumen. PEG treated forage has greater rapidly soluble fraction (a), 

PEG treatment can not increase potentially degradable fraction (b), 

but NaoH solution was done. Effective degradability in different 

rumen dilution rates were high for PEG treated Sainfoin compared to 

others. 

Treatment a (%) b(%) C(h -1 ) K=0.03 K=0.05 K=0.08 

(h -1 ) (h -1 ) (h -1 ) 

PEG 47.7 a 32.8 ab 0.076 a 69.03 a 64.53 a 60.06 a 

NaHCO 3 37.4 b 30.17 b 0.076 a 56.53 c 52.26 c 48.06 c 

NaoH 35.9 c 37.0 a 0.08 a 61.2b 56.5 b 51.73 b 

Control 33.4 d 36.35 a 0.066 a 57.2b c 54b c 49.9b c 

Treatment Rumen incubation time(h) 

4 8 12 24 48 72 96 

Control 40.71 b 49.37 b 51.52 b 62 c 67.43 b 69.25 bc 70.22 bc 

NaHCO 3 39.03 b 43.8 c 50.06 b 62.92 c 66.4 b 67.04 c 67.13 c 

NaoH 42.64 b 50.13 b 53.94 b 68.86 b 70.6 b 72.50 b 73.08 b 

PEG 50.36 a 56.88 a 62.16 a 75.56 a 79.34 a 80.01 a 80.15 a

Means within a column with different superscripts differ. (p

equal distributions at 08:00 and 16:00 hours. This diet met energy 

and protein requirements according to CNCPS standards. On the last 

day of each period, blood samples were taken at 5h after feeding 

from the jugular vein of each sheep, alternating sides at each 

collection. The blood was collected in a 10-ml Vacutainer tube 

(contain heparin), and blood pH and CO 2 were analyzed within 1h. 

Blood samples centrifuged at 3000 xg for 20 min and plasma was 

used to analyze glucose and lactate dehyrogenase (LDH). DFM 

treatments had no effect on blood variables (Table 1).Table 

1.Characteristics of blood parameters in sheep fed treatments 

Item 1 a 2 b 3 c 4 d SE e 

CO 2 43.8 44.23 44.15 45.18 0.29 

pH 7.3 7.32 7.29 7.34 0.02 

LDH 378.44 381.16 395.29 401.32 39.6 

Glucose 81.41 80.13 82.81 81.1 3.1 

a control contain alfalfa hay as Source of roughage, b control contain corn silage 

as Source of roughage, c diet 1 with DFM, d diet 2 with DFM, e standard error. 

1078 Effects of Alfalfa Hay Particle Size and Cottonseed Hulls as 

Nonforage Fiber Source on Performance of Holstein Dairy 

Cows 

H. Abdi benemar, K. Rezayazdi, A. Nikkhah 

University Of Tehran, Animal Science Department, Karaj, Tehran, 

Iran 

Introduction: Ration particle size has been observed to affect DMI, 

milk fat and protein content and milk yield. Nonforage fiber sources 

(NFFS) possess a large amount of NDF that can be used as a forage 

substitute based on their price and availability. Cottonseed hulls (CSH), 

a by-product of cotton processing, contain a large proportion of NDF 

and associated lignin, and have been considered a useful NFFS. It was 

suggested that physical characteristics of dietary forage can interact 

with nonforage fiber sources. The objectives of this study were to 

determine effects of CSH as nonforage fiber sources and to investigate 

interaction between CSH as NFFS and alfalfa particle size on DMI, 

milk fat and protein content and milk yield. 

Materials and methods: Twelve Holstein dairy cows in 

midlactation were assigned to a change-over design with three 

periods and 2_2 factorial arrangement of treatments. During each of 

28-d periods, cows were offered one of four total mixed rations that 

differed in alfalfa hay particle length (long and short) and cottonseed 

hulls substituted for AH (0 and 9.63% DM. Cows were housed in tie 

stalls and fed twice daily at 0800 and 1400 h for adlibitum 

consumption. Milk yield was recorded twice weekly and milk 

samples analyzed for fat and protein content. Data were analyzed 

using the mixed model procedure of SAS (1998) with model effects 

for period, alfalfa hay particle length (APL), fiber source (FS) and 

two-way interaction of APL by FS as fixed effects and cow as 

random effect and effects of factors were declared at P < 0.05. 

Results: Dry matter intake was not affected by either APL or 

substitution of CSH. Milk yield, 4% FCM, and percent of milk fat and 

milk protein were not affected by either APS or substitution of CSH 

Table 1. Effects of alfalfa hay particle size (APS) and fiber source (FS) 

on milk production, composition and dry matter intake 

Treatments Effects 

LGNH LGH SHNH SHH SEM APL FS I 1 

Milk yield, Kg/d 26.57 28.46 28.00 28.22 1.17 0.48 0.21 0.32 

4% FCM, Kg/d 25.16 26.64 26.35 26.52 1.23 0.54 0.35 0.46 

Fat. % 3.24 3.16 3.20 3.17 0.16 0.88 0.70 0.97 

Protein, % 3.10 3.03 3.13 3.26 0.12 0.14 0.72 0.25 

PeNDF, % 2 26.29 32.21 24.29 30.17 2.03 * ** 0.98 

DMI, Kg/d 23.48 25.11 24.94 25.89 1.17 0.48 0.21 0.42 

Conclusion: The results of this study showed that cottonseed hulls can 

be substituted partly for alfalfa hay when forage sources are expensive 

or its stores are limiting. 

Key words: particle size, alfalfa hay, cottonseed hulls, dairy cows 

1079 Concentration of Conjugated Linoleic Acid (CLA) and other 

Fatty Acids in Parmigiano-Reggiano Cheese 


A. Formigoni, N. Brogna, F. Merendi, A. Mordenti, G. Biagi 

University of Bologna, DIMORFIPA, Ozzano emilia, Italy 

Conjugated linoleic acid (CLA) is a nutrient that can exert several 

beneficial effects on the health of consumers and is mainly found in 

meat and dairy products from ruminants. In order to characterize the 

lipid fraction of Parmigiano-Reggiano cheese, 255 samples of 

cheese were analyzed. Samples differed for geographical origin 

(different provinces constituting the Parmigiano-Reggiano cheese 

area of production), altitude of the cheese factory location (plain, 

hills or mountains; P, H or M), and seasoning time (< 22 months, 

between 22 and 30 months, or > 30 months). All samples were 

analyzed for their lipid content and fatty acids composition. Total 

lipid content of cheese was not influenced by any of the variables 

considered and averaged 24.4 %. Content of all fatty acids was not 

significantly affected by seasoning time. In particular, content of 

CLA averaged 0.32 mg/100 mg of fat. The altitude of the cheese 

factory location had a significant influence on the concentration of 

several fatty acids. Cheese samples coming from factories located on 

mountains showed significantly (P < 0.05) higher concentrations of 

C18:0 (stearic acid; 6.1 vs 5.5 and 5.3 mg/100 mg of fat for M, H and 

P, respectively), C18:1 cis (oleic acid; 13.0 vs 11.5 and 11.5 mg/100 

mg of fat for M, H and P, respectively), C18:1 trans (vaccenic acid; 

1.04 vs 0.89 and 0.76 mg/100 mg of fat for M, H and P, 

respectively), and CLA (0.37 vs 0.32 and 0.28 mg/100 mg of fat for 

M, H and P, respectively). Similarly, the concentration of some fatty 

acids was influenced by the province of origin, but this effect seemed 

to be mainly related to the different presence of cheese factories 

located on mountains in each province. The present results show that 

the area of production of Parmigiano-Reggiano cheese significantly 

influences its lipid fraction. In particular, cheese from factories 

located on mountains show a higher content of CLA and vaccenic 

acid, probably due to a higher utilization of green forages which are 

very rich in linoleic and ·-linolenic acid, both precursors of CLA. 

Key words: dairy cow, Parmigiano-Reggiano cheese, conjugated 

linoleic acid, fatty acids 

1235 Adrenal Weight and Morphology in Different Ages of Cow 

A. Mohammadpour 

Ferdowsi University of Mashhad, Faculty of Veterinary Medicine, 

Department of Anatomical Sciences, Mashhad, Iran 

Eighty pair of adrenal glands in Holstein breed cow was 

histomorphometrically studied in four different age groups included: 6 

month to 9 month, 9 - 12 month, 12 - 15 month and 15 - 18 month ages. 

After collecting the glands and weighting them, some parameters such as: 

length, wide and thickness of left and right adrenal were measured by 

calliper device. For histological studies, after tissue preparation and 

staining with H&E, cortical and medullar zones were measured. At 

results, data between left and right adrenal glands in the four ages were 

analyzed and compared using ANOVA and t-student tests. In Holstein 

cow the left adrenal gland was been C shape and right adrenal gland was 

been V shape. There was significant difference between dimensions of left 

and right adrenal glands. The mean weight, length, wide and thickness of 

left adrenal gland were greater than right in four ages. In comparison 

between different age groups there was a significant difference in all of 

parameters between ages 6 to 9 month and 15 - 18 month (P

cows mainly depends upon roughage, straw etc. that provide mostly 

carbohydrate. Fodders like cereal grains, rice polish etc. are highly 

expensive and beyond the reach of marginal farmers, so they are 

unable to supply these to their cows and their cows suffer from 

protein, mineral and vitamin deficiency. Iodine is necessary for 

thyroxin production that has a positive effect on milk yield. 

Considering it this experiment was conducted to find out a cheapest 

way of increasing milk yield. A total of 20 indigenous dairy cows of 

about 3-5 years old, weighing from 120-190 kg were selected based 

upon history at the day of parturition and were randomly divided into 

4 groups. Group A acted as control, and Group B, C and D were 

treated with non-iodide salt at 100 mg/kg bd. wt. PO daily once, 

iodide salt at 100 mg/kg bd. wt. PO daily once and Lugol’s iodine at 

10 mg/kg bd. wt. IV twice a month, respectively. All the cows were 

maintained in a similar fashion. Milk yield were recorded daily for 5 

months and all the data were analyzed statistically at the end of 

experiment. Significant (p

electrochemical properties of the analyte under analysis. Recent studies 

have demonstrated the capacity of these devices to detect and quantify 

micro- and submicro molar concentrations of both redox mediators (id. 

Metal-organic compounds), commonly used in electrochemical 

biosensors, and other molecules. The applications of these devices in 

Veterinary Medicine are: quantitative determination of drugs, steroids, 

pathogenic bacteria, toxins, Genetically Modified Organisms, 

Polychlorinated biphenils, and pesticides. In this study the copper ion 

(Cu 2+ ), has been determined as scientific literature reports both high 

serum Cu concentrations in molluscicide polluted water intoxication in 

bovine and ovine species, and low serum concentrations in calves with 

neuromuscular problems. An electrochemical nano-structured 

biosensor, with both high ratio Signal/Noise (S/N), and very high 

selectivity, has been utilised for determination of Cu 2+ in a water 

matrix. 

Materials and methods: A nano-structured electrochemical 

biosensor, ultra sensitive and highly accurate with a wide spectrum of 

application, was planned and developed in 2005-2007 years by Nano- 

SIMS s.r.l. by Nano Imprinting Lithography (NIL) on polymeric 

substrate, and utilised as a working electrode in electro analytical 

technique of cyclic voltammetery. This biosensor was used for ultra 

trace quantitative determination of Cu 2+ in water matrix has been 

developed. 

Results: This method, using a specific Cu biosensor, has allowed the 

determination of a very low Cu concentration of 1,2 x 10 7 ions/nL in 

water matrix. 

Conclusions: The development of this technology allows both the 

specific chemical programming of the biosensor, to assure the very 

high selectivity and specificity of the analyte to be determined, as 

well as the use of the biosensor in low cost devices for field 

applications. It is important to highlight the usage of this biosensor 

both regarding quality and safety of food and mass screening, and for 

the ultra trace quantitative determination of numerous other 

analytes, and for notable potential applications in Veterinary 

Medicine which will be studied in detail, in the near future, 

particularly in bovine and ovine species. 

Key words: nanotechnology, biosensor, copper, bovine and ovine 

species. 

1242 Diagnostic Values of Haptoglobin and Serum Amyloid a 

Concentrations in Bovine Heart Diseases 

A. Rezakhani, S. Nazifi, A. Moaddeli, M. Zarifi 

School of Veterinary Medicine, Shiraz University, Clinical Sciences, 

Shiraz, Iran 

Congenital and acquired heart diseases have been reported in cattle. 

Acquired cardiac problems can be manifested as murmurs or 

arrhythmias. The prevalence of cardiac murmurs and arrhythmias 

among clinically healthy dairy cattle has been under investigation in 

our school for the past 5 years. The prevalence of valvular cysts 

which can produce cardiac murmurs has also been reported in our 

institution. Various methods of diagnostic tools have been used for 

diagnosis of heart diseases in animals. This study was carried out to 

find a possible relationship between cardiac murmurs and the 

concentration of acute phase proteins (haptoglobin and serum 

amyloid A) in dairy cattle population. Two hundred dairy cattle of 

different ages from four dairy farms were examined by auscultation 

on the base of method used by Rezakhani and Zarifi (2007). 77 out 

of these 200 cattle had various types of cardiac murmurs. Cases of 

pericarditis and endocarditis referred to our school were taken into 

this study for comparison. Twenty normal cattle without any sign of 

heart disease and cardiac murmurs were taken as a control group. 

Blood samples were taken from all cattle with cardiac murmurs, 

from cases referred to school teaching hospital and from the control 

group for determination of haptoglobin and serum amyloid A plus 

CBC. Statistical analysis carried out on the findings of this study 

showed that there was significant difference in the concentration of 

these two acute phase proteins in endocarditis and pericarditis group 

as compared with the control group and the clinically normal cattle 

with cardiac murmurs (P 0.05). The findings also showed that 

there was significant difference in CBC in cases with enocarditis and 

pericarditis as compared with group having cardiac murmurs and the 

control group (P< 0.05). This study showed that acute phase proteins 


measurement can not be used for the differentiation of functional 

from pathological murmurs. 

Key words: heart diseases, murmurs, cattle, haptoglobin, serum 

amyloid A 

1243 Auditory Evoked Response in the Clinically Normal Cows: 

Middle Latency Components 

A. Pomianowski, Z. Kuleta, M. Kwiatkowska 



Objectives of study: Our objective was to record the middle latency 

components (10 to 60 ms) of the auditory-evoked response in clinically 

normal mature dairy cows. 

Materials and methods: To detect the middle latency response 

(MLR), a series of suprathreshold tone pips was presented monoaurally 

to nonanesthetized cows (n = 6 mixed breed) at a stimulus frequency 1 

kHz. Stimulus intensity tone pips were presented at 80 dB hearing level 

(nHL), introduced through insert earphone tranducer. Contralateral 

masking noise at 50 dB nHL was used. Filtered electroencephalic 

activity during the first 100 ms after each tone pip stimulation was 

recorded and averaged digitally for 2000 consecutive stimuli. 

Results: The response pattern in the cows was repeatable polyphasic 

configuration characterized by 4 major waves. Peaks of the waves were 

labelled, using a method developed in human beings. Mean latency 

ranges for the positive (P) or negative (N) peaks were 11,2 to 12,6 ms 

(Na), 17,5 to 18,3 ms (Pa), 27,1 to 28,8 ms (Nb) and 43,4 to 45,4 ms 

(Pb). 

Conclusions: Configuration of used parameters allows to detect MLR 

in cows. They reflect the functional integrity of the thalamus-cortical 

region and primary auditory cortex. Measurement of peak latencies 

provides information to supplement the results of standard neurologic 

examination and can be useful tool for monitoring anaesthesia and 

analgesia. 

Key words: middle latency responses, cows 

1244 A Simplified Method of Estimation of Plasma Fibrinogen 

Concentration without Heat Precipitation 

A. Roussel 1 , J. Osterstock 1 , A. Bouchant 1 , E. Calzamiglia 1 , 

K. Russell 2 

1 

Texas A&M University, Large Animal Clinical Sciences, College 

Station, United States 

2 

Texas A&M University, Veterinary Pathobiology, College Station, 

United States 

Plasma fibrinogen concentration (FC) determined by heat 

precipitation (HP) has been used to support the diagnosis of 

inflammatory disease in cattle. This method requires a heated water 

bath which is not universally available in practice. Our objective was 

to evaluate an alternative method of FC measurement which does not 

require HP. We studied 75 cattle (sick and clinically normal) 

admitted to the Texas Veterinary Medical Center. Blood samples 

were collected into plain vacuum tubes and vacuum tubes containing 

EDTA. Blood from the EDTA-containing tubes was centrifuged for 

4 minutes in 2 micro-hematocrit tubes within 60 minutes of 

collection, and the plasma protein concentration was estimated from 

one of the tubes using light refractometry. The second microhematocrit 

tube was placed in a water bath heated to 56° C for 5 

minutes to precipitate the fibrinogen and then centrifuged. The 

serum protein concentration on this sample was estimated using 

refractometry. The difference in the protein concentration between 

the two micro-hematocrit tubes is the FC determined by HP. After 1 

hour at ambient indoor temperature, the plain tube was centrifuged, 

and serum protein concentration was estimated using refractometry. 

The FC was then estimated by subtracting the serum protein 

concentration from the plasma protein concentration. Summary 

statistics were calculated and means were compared by method 

using a paired t-test and by operator using a t-test. The FC was also 

compared between the 2 methods using linear mixed-effects models. 

Mean FC from HP was 441 mg/dl (S.D. 142.0 mg/dl) and mean FC 

from ambient temperature clotting was 412 mg/dl (S.D. 155.4 

mg/dl). The difference between the FC measured by HP and ambient 

temperature clotting was 0 for 22 cattle, 100 mg/dl for 42 cattle and 

200 mg/dl for 11 cattle. Comparison of results obtained using HP 

between different operators revealed a significant difference

etween mean results for the 2 operators in this study (P = 0.003). 

Results from the mixed-effects model indicated that results of the 

two methods were not significantly different (P = 0.66). However, a 

significant effect associated with operator (P = 0.04) and a 

significant method-by-operator interaction (P = 0.02) were detected. 

The results of this model demonstrate that estimation of FC using HP 

and the ambient temperature clotting yields similar results, but 

differences in the results obtained by different operators may affect 

results of the HP method. 

1245 Seasonal and Regional Differences of Iodine Concentration 

in Raw Cow, Sheep, and Goat Milk in Slovakia 

I. Paulikova, H. Seidel, O. Nagy, G. Kovac 

University of Veterinary Medicine, Kosice, Slovakia 

Aim of this work was to map iodine content in raw milk of dairy 

cows, sheep, and goats regardless breed, production, and 

reproduction cycle and to compare milk iodine levels in various 

Slovakian regions, as well milk iodine concentrations during winter 

and summer feeding periods. The samples were taken manually after 

washing of quarters and halves, resp., without use of iodine 

disinfectant. The animals from 32 farms (dairy cows - 23, sheep - 7, 

goats - 2) were divided into four groups according to the regions – 

Western, Middle, Eastern, and Northern Slovakia. We compared 

also milk iodine concentrations recorded during the summer (1 May 

- 31 October) and winter (1 November - 30 April) feeding periods. 

The milk iodine concentrations were determined by photometric 

method for analysis of trace iodine based on catalytic reaction NO 2 - 

/SCN - . Milk iodine concentrations were determined in 457 samples 

of raw milk from dairy cows, 79 samples of sheep, and 17 samples of 

goat milk collected in various Slovakian regions from 2002 to 2007. 

Among all the 457 samples of bovine milk, iodine content below 50 

g.l -1 was recorded in 113 samples (29.94 %); 294 samples (64.33 %) 

ranged between 50 and 200 g.l -1 ; 19 samples (4.16 %) from 200 to 

500 g.l -1 ; 17 samples (3.72 %) between 500 and 1000 g.l -1 , and 13 

samples (2.85 %) showed milk iodine concentrations over 1000 g.l - 

1 . Concentrations in the Slovakian regions showed the highest values 

in the Western, then Middle and Eastern Slovakia, and the lowest 

values in the Northern Slovakia (p < 0.05, p < 0.01). However, the 

results showed large variability within both regions and farms. In 

sheep and goat milk samples, we found milk iodine concentrations 

below 80 g.l -1 in 49 sheep (62 %) and in 6 goats below 60 g.l -1 (37.5 

%), which are indicative of iodine deficiency. When comparing 

seasonal differences, sheep and goat milk was higher in the iodine 

content during the winter feeding period, however, in dairy cows we 

recorded opposite ratio. Except goat milk (p < 0.01) these difference 

were not significant. 

Key words: iodine, raw milk, cattle, sheep, goat 

1246 Family Dairy Production Process in Mexico - a Case Study 

R. J. Jiménez 1 , F. P. Alonso 1 , L. H. García 2 , V. O. Espinosa 1 , 

A. P. Alonso 1 , L. P. Brunett 3 , J. G. Meléndez 1 , J. T. Cruz 1 

1 

Facultad de Medicina Veterinaria y Zootecnia - UNAM, 

Departamento de Economía, Administración y Desarrollo Rural, 

Mexico City, Mexico 

2 


Producción Agrícola y Animal, Mexico City, Mexico 

3 

UAEM, Facultad de Medicina Veterinaria y Zootecnia, Mexico 

State, Mexico 

The aim of this project was to describe the family dairy process 

production in order to find proposals to improve the production of 

this system. The project was made in 20-production units in four 

communities of Maravatio, Michocan, Mexico. Through 

questionnaires, semi-structured interviews and participative 

observation was gotten information about the productive process. It 

was made a frequencies study and the activities of the process were 

described. Results obtained showed that cow ownership was 

influenced mainly by Holstein-Friesian cross. They have herds less 

to 20 cows that are maintained in semi-extensive system in summer 

and feed-lot system in winter. In technology, all of them have 

preventive medicine schedules; 14 producers performed artificial 

insemination, and only 7 producers have mechanical milking. In the 

feeding mainly are used holding crop residues, maize, sorghum, 

commercial balanced food and grazing. Family labour is used on 

production process in 95%. Four main activities were identified: 

facilities cleaning, milking, milk delivery and feeding. It was 

observed improper techniques in: 1) the manual milking that have a 

negative influence on the quality product; 2) the feeding, due to the 

inefficient use of resources; and 3) the marketing because of the 

entrepreneurs who reduced the producers’ income. However, some 

producers have adapted technology which let them to improve their 

system. The competitiveness and permanence in a global market is 

based on the quality product and the efficient use of sources. 

Furthermore, the Maravatio’s family diary production has both 

weakness, if it does not improve, it would be out of commercial 

market. 

This work was supported by Program for technological Research and 

Innovation Projects of the National autonomous University of Mexico 

(PAPIIT IN304307). 

Key words: family dairy, Mexico, process production 

1247 Effect of Three Different Phytotherapics on Bovine Skin 

Wound Healing Process 

L. Lipinski 1 , A. Wouk 2 , N. Lemos da Silva 3 , D. Perotto 3 , R. Ollhoff 4 

1 Universidade Federal do Paraná, Veterinary Clinical Sciences, 

Curitiba, Brazil 

2 Universidade Federal do Paraná, Veterinary Clinical Sciences, 

Curitiba, Brazil 

3 Instituto Agronômico do Paraná, Ponta Grossa, Brazil 

4 Pontifícia Universidade Católica do Paraná, Ciencia Animal, Sao 

José dos Pinhais, Brazil 

Objectives: The present study aimed to compare the effect of the 

bark of Schinus therebentifolius, the sawdust of Tabebuia 

avellanedae and leaves and branches of Caesaria sylvestris, all 

native brazilian trees, on the wound healing process of the bovine 

skin by second intention. 

Material and methods: The skin of 11 Purun heifers was surgically 

removed with a 2cm - punch on 4 locations on both sides of the lombar 

region, on a parallel line 8 cm from the vertebral column and treated 

topically daily for 17 days with the decoctus of the three plants. One 

lesion on each side remained as control and was rinsed with saline 

solution. One row was randomly chosen to be used for the 

histopathological excisions. The lesion was macroscopically evaluated 

at 5 times and the microscopic aspect (inflammatory response, young 

and mature granulation tissue) was evaluated twice (day 6 and 16). The 

centripetal cicatrization was measured drawing the border line of the 

lesion on a transparent sheet and calculating the area beneath. All 

animals were kept in individual stables with a previous 10 days 

adaption and received the same balanced feed. The macroscopic aspect 

was judged always by the same person who applied 3 different degrees 

(1 = bad, 2 = regular, 3 = good), considering local hemorrhage, 

presence of coagula, crusts, granulation tissue, epithelialization, 

presence of exudate. 

Results and discussion: Lesions treated with Tabebuia were better 

evaluated probably due to his anti-inflammatory properties, although 

histopathology with all treatments showed no influence over 

inflammation. Lesions treated with Caesaria initially did not differ 

from Schinus, but could not be differentiated subsequently from the 

control. Lesions treated with Schinus had a more dry and irregular 

aspect, probably due to the effect of tannines. The cicatrization area did 

not differ at any time from the control, but Tabebuia treated lesions 

showed a statistically measured tendency of faster improvement. The 

histopathological examination revealed a positive effect of Tabebuia 

and Schinus over fibroplasia. 

Conclusion: The phytoterapic extracted from Tabebuia avellanedae 

and Schinus therebentifolius improved cicatrization whereas Caesaria 

sylvestris showed a beneficial effect only at the beginning of the 

healing process. Phytoterapics could be used to promote the healing 

process as an alternative to disinfectants i.e. in organic farming 

systems. 

Key words: phytotherapy, wound, healing, bovine 

1248 Selenium Profile in Blood and Hair from Cattle, 

Environmentally Exposed to Selenium around Isfahan 

Ceramic Industry, Iran 

M. Pourjafar 1 , H. Sateri 2 

1 Shahrekord University, Clinical Sciences, Shahrekord, Iran 

2 Islamic Azad University of Shahrekord, Shahrekord, Iran 

Miscellaneous 309

Selenium was discovered as an essential nutrient in 1957, though it 

was not discovered what role it played in the body until 1973. There 

is more data on the toxicity of Se in cattle. Selenium may be stored 

in liver, kidneys, blood and hair chronically. This survey aimed at 

acquaintance the amount of dairy farms selenium pollution around 

the Isfahan ceramic industry. At first, location of the dairy farms 

around Isfahan ceramic industry was identified. Then dairy farms 

that were in distant less than 2.5km identified and divided in three 

groups. Group A: farms located in radial zone under 1 Km. Group B: 

farms located in radial zone of 1 to 2 Km. Group C: farms located in 

radial zone of 2-3 Km. Group D (control farms) were in nonpolluting 

places of Isfahan. From cows in each group blood and hair samples 

were collected. The selenium content of samples was measured by 

Atomic Absorption Spectrometer. The cows which were located in 

1-2 Km group to ceramic industry had higher blood and hair 

selenium content and statistically significant differences exist 

between other both groups and control group(P

eport a case of a hepatocellular carcinoma in a 10 years old cow, 

Tabapu’ breed in Brazil. Gross pathology of the liver revealed a 

solitary protruded tumor above the surface of an entire lobe and 

metastasis to the visceral peritoneum of rumen, reticulum, omasum, 

abomasum, mesentery and lymph nodes. Tumors were friable and 

yellowish in color. Histologically, the intrahepatic tumor had a solid 

pattern composed by atypical hepatocytes surrounded by a fibrous 

capsule, and the mass compressed normal hepatic parenchyma. 

Metastasis in the visceral peritoneum and mesentery showed a 

pseudoglandular pattern. The tumors revealed intense cellular and 

nuclear anaplasia. These gross and histopathological findings are 

consistent with the diagnosis of hepatocellular carcinoma, which so 

far was not described in Tabapu’ cows. 

Sponsored by: FAPESP - Brazil 

Key words: hepatocellular carcinoma, bovine, gross lesions, 

histopathology 

1253 Phonocardiographical Investigations of Heart Murmurs in 

Cattle 

A. Reisse, M. Metzner, G. Rademacher, C. Sauter-Louis, W. Klee 

University of Munich, Clinic for Ruminants, Munich, Germany 

In cattle medicine, heart murmurs are usually detected by heart 

auscultation. The aim of the present study was to examine the 

feasibility of describing heart murmurs using phonocardiography. 

The objective of the study is to establish an objective description of 

cardiac murmurs using phonocardiography. 20 Simmental cows, 

patients of the Clinic for Ruminants, University Munich, selected 

randomly were examined. The cows received a standardised 

examination including heart auscultation. Commonly, cardiac 

murmurs are divided into six degrees. Depending on the results of 

the heart auscultation, the animals were classified 

accordingly. Phonocardiographic records were taken for each animal 

at the third and fourth intercostal spaces (ICS) at a dorsal and ventral 

location on each side.For each location three consecutive, equal 

cardiac actions (c.a.) were selected in a diagram of 50 and 100 Hz. 

Cut-offs for each location, each frequency (50 and 100 Hz) and for 

the negative and positive amplitudes had been established in a pilot 

project. These cut-off values were used to distinguish between 

background noise and potential heart murmurs. A simultaneously 

recorded electrocardiogram was used as a time control to divide c.a. 

into systole and diastole. Both periods were divided thereafter into 

equal intervals of 10 (50 Hz) and 20 (100 Hz). The heights of the 

amplitudes at each of these points were checked against the chosen 

cut-offs. The area under the curve (AUC) was calculated for each 

diagram. Data were analysed in conjunction with the clinical 

findings, the grade of the cardiac murmur, and laboratory 

examination parameters. Preliminary results, there were differences 

in the frequency of the time points, at which the amplitudes were 

outside the set limits, between animals with faint cardiac murmurs 

(degree 1 or 2) and animals with severe cardiac murmurs (degree 5 

or 6). Animals with faint murmurs showed fewer amplitudes outside 

the limits in the systole in the 3. ICS on the left side than animals 

with severe murmurs (p = 0.021 at 50 Hz, p = 0.014 at 100 Hz). 

These differences were also visible in the AUC between these two 

groups. At 50 Hz the AUC for the systole was significantly smaller 

in animals with faint murmurs than in animals with severe murmurs 

(p = 0.037) at the 3. ICS on the left side. In further studies, a scheme 

for the analysis of echocardiography will be developed, which will 

allow an objective assessment of heart sounds and murmurs in cattle. 

1254 Evaluation of the Immunogenic Capacity of a Clostridium 

chauvoei Recombinant Protein for Prevention of Blackleg 

L. Ontiveros, L. Hernandez, J. Lopez, V. Tenorio 

INIFAP, Bacteriologia, México DF, Mexico 

Diseases in livestock caused by Clostridium spp are very important 

in Mexico. There are not vaccines of good quality against these 

infections; for this reason several outbreaks have occurred in recent 

years. The objective of this work was to study the immunogenic 

capacity of a recombinant protein of 156 kDa of Clostridium 

chauvoei that has shown 80% protection against this disease in 

guinea pigs. This immunogenic protein was cloned in the expression 

vector pBluescript and was used to immunize Clostridium chauvoei 

free bovines that were kept in an endemic area. Three experimental 

groups were studied. In group 1, 30 bovines were vaccinated by 

subcutaneous route with one dose of 350 ug/animal of the 

recombinant protein of 156 kDa. In group 2, 30 bovines were 

vaccinated with the same concentration of this protein plus 

aluminium hydroxide as adjuvant. Group 3 was vaccinated with a 

commercial bacterin by intramuscular route with a dose of 

5ml/animal. In each group 5 animals were inoculated with saline 

solution and remained as controls without vaccination. Blood 

samples were obtained each month during six months. Serum 

samples were analyzed by agglutination test and Western blotting. 

The recombinant protein of 156 kDa was recognized by serum 

samples from all the animals in groups 1 and 2. Only 2 animals from 

group 3 recognize this protein. During the time of the experiment 

any cases of this disease were observed. However, longer time or 

stress conditions that would favour occurrence of the disease would 

be required to confirm if this immunogen is also protective in 

bovines 

Key words: Clostridium chauvoei, blackleg, bovine 

1255 Inhibition of VPIB-3 Virus for Hemagglutinant Fraction 

Isolated from Crotalid Venom 

A. Alvarado 1 , Y. León 2 , O. De Paz 1 , C. Gonzalez 3 , E. Hernández 3 , 

A. Aguilar 2 

1 INIFAP, Virology, México DF, Mexico 

2 Instituto Nacional de Pediatría, Immunology, México DF, Mexico 

3 Facultad de Medicina Veterinaria y Zootecnia-UNAM, Department 

of Pathology, México DF, Mexico 

An hemagglutinant fraction antiviral activity of crotalid venom 

against Parainfluenza virus (VPIB-3) was evaluated from crotalid 

serpents; Agkistrodon piscivorus, Agkistrodon contortix and 

Crotalus scutulatus. Agkistrodon piscivorus venom showed the 

greatest hemagglutinant capacity with 64 units. The nonlethal 

maximum permissible IM dose of the three venoms was 1.75 mg/30g 

in mouse. Agkistrodon piscivorus and Agkistrodon contortix caused 

massive edema and sanguineous infiltration in muscle; microscopic 

lesions were acute miositis in various severitu degrees with the three 

different venoms. Loosening of monolayer or agglutination in 

MDBK cells was observed with each one of venoms in doses higher 

than 0.006, 0.012 and 0.195 mg/millilitre, respectively. The highest 

hemagglutinant activity fraction was isolated from Agkistrodon 

piscivorus venom bye HPLC-RP. Mice IM inoculation with this 

fraction (4.25 µg/0.2 millilitre), caused no macroscopic or 

microscopic damage and MDBK cells metabolism was no altered 

with concentrations bellow 1.062 µg/millilitre. Also the 

hamagglutinant fraction prevented the infection with VPIB-3, and 

kept between 63.34 and 84.66% the cellular viability. On the other 

hand, it was observed (electron microscope), that hemagglutinant 

fraction destroyed the viral particles. Finally, a common protein 

(20.99 kDa) to Agkistrodon piscivorus venom and VPIB-3 was 

recognized by an anti-venom serum. The results show that the 

purification of the hamagglutinant fraction of the venom, eliminates 

the damage caused by the others components, making possible their 

use like antiviral compound. 

Key words: crotalid fraction, VPIB-3, lectins, hemagglutinin 

1256 Effect of Diet Supplementation with Aromatic Plants in 

Performance and Meat Quality of Fattening Cattle 

G. Arsenos, P. Fortomaris, A. Yannakopoulos, A. Tserveni-Gousi 

Aristotle University of Thessaloniki, Animal Production, Thessaloniki, 

Greece 

The objectives were to assess the extent to which the performance of 

fattening cattle, as well as the quality of the meat produced, could be 

affected by the incorporation in the diet of aromatic plants for a 

period of two months prior to slaughter. A total of 32 entire male 

Limusine bulls were used. At the commencement of the experiment 

their average body weight (BW) was 522±21.7 kg (mean SD) and 

they were allocated into two groups (n=16); A (aromatic) and C 

(control), balanced for BW. They were kept in feedlot pens and 

offered a basal ration comprised of concentrates and straw both 

offered ad libitum. Two months before the designated day of 

slaughter a mixture of aromatic plants was incorporated in the ration 

of group A, at a level of 8%, whereas group C was offered the basal 

ration. Feed intake was calculated from refusals measured daily and 

Miscellaneous 311

growth by measuring BW at 15-day intervals. All animals were 

slaughtered at the same day in an EU registered slaughterhouse. 

Their carcasses were assessed for conformation and scored 

according to the EU standard scheme. After obtaining hot carcass 

weight the carcasses were kept for six hours at room temperature 

before refrigeration at +1C for 24 h. Subsequently, the carcasses 

were re-weighed (cold carcass weight) and the ultimate pH (pHu) 

measured in the longissimus dorsi. Thereafter, eight (8) carcasses 

from each group were randomly selected for comprehensive meat 

quality measurements: (i) CIE colour space was used to measure 

meat colour in the muscle longissimus dorsi, (i) meat moisture, total 

fat, and protein content and (iii) fatty acid composition of muscle 

tissue. The results showed that there were not significant differences 

in the BW of cattle at slaughter (631 vs 612, s.e.d 13.2 for groups A 

and C, respectively. However, there were significant differences 

(P

correlations between Occurrence of ocular squamous cell carcinoma 

and sex and any clinical problems in the studied animals. Cattle with 

unpigmented or partly pigmented eyelids were at most risk. 

Key words: bovine ocular squamous cell carcinoma, Holstein cattle, 

Iran 

1261 Serum Cardiac Troponin I Concentrations as a Diagnostic 

Aid of Pericarditis in Cattle 

H. Guyot 1 , A. Sartelet 1 , K. Leidinger 2 , S. Vandeputte 1 , F. Rollin 1 , 

H. Amory 1 , C. Sandersen 1 

1 

University of Liege - Faculty of Veterinary Medicine, Department of 

Clinical Sciences, Liege, Belgium 

2 

Bicontrol, Department of Clinical Sciences, Ingelheim am Rhein, 

Germany 

Cardiac troponin I (cTnI), a specific marker of myocardial damage, 

is widely used in the man and of increasing importance in veterinary 

medicine. It is not commonly used in cattle practice although it 

might be useful as a diagnostic aid of pericarditis. Pericarditis is a 

common complication of traumatic reticuloperitonitis in cattle but 

clinical signs are non-specific and diagnostic aids are not readily 

available in field conditions. The aim of the study is to test the 

usefulness of cTnI as a blood marker of pericarditis in cattle. 

Nineteen cows of various breeds with clinical signs of pericarditis 

were compared to a control group composed of 8 age and breedmatched 

cows. Serum cTnI levels were measured in all cows by a 

direct chemiluminescence sandwich-immunoassay using one 

polyclonal goat-anti cTnI and two monoclonal mouse anti-TnI antibodies 

(ADVIA Centaur TnI-Ultra-Test, Biocontrol, Ingelheim, 

Germany). Pericarditis was confirmed by typical echocardiographic 

images and/or post-mortem examination. Statistical analysis was 

performed using non-parametric Mann-Whitney Wilcoxon test. 

Cardiac TnI levels of control group cows were below 0.04 ng/mL. 

Five of the pericarditis-affected cows had cTnI levels of 0.04 ng/mL 

or below. Fourteen pericarditis-affected cows had serum cTnI levels 

from 0.06 to 5.10 ng/mL. The sensitivity of the optimal cut-off value 

of 0.04 ng/mL was 74% (95% C.I., 54% to 93%), specificity 100% 

(95% C.I., 100% to 100%), positive predictive value 100% (95% 

C.I., 100% to 100%), negative predictive value 62% (95% C.I., 35% 

to 88%). The cTnI blood concentration was significantly higher 

(p35 L), the VWP was 

maximum 90 days and cows were synchronized using the PGF2· or 

the OvSynch-protocol. After excluding the slaughtered animals and 

those, who did not become pregnant during the time of the 

experiment, 76 (G: A) and 83 (G: B) animals were evaluated. 

Conception rate of the first service (CR) was 31.6% in G: A and 

38.6% in G: B. At the same time, in SG II and IV it was 14.8% and 

45.8% for G:A and 42.8% and 50% for G: B. According to the 

applied different VWP, the duration until first AI was longer for 

G:B (94.9 days) than for G:A (88.5 days), while the calving to 

conception interval (CCI) became shorter for G: B (156.4 days) than 

for G: A (168.2 days). Also less services per conception and total 

number of inseminations were needed for G: B (2.58 and 212) than 

for G: A (2.98 and 220), respectively, although G: B required more 

costs for hormonal treatments. In general, there was a 12-day 

difference in the CCI, but none of the results showed any significant 

differences. As conclusion, US turned out to be a useful tool in 

reproductive management of large dairy herds because by using it, 

we can increase the accuracy of diagnoses. These results may 

suggest that by setting up individual VWP-s based on the actual 

daily milk yield and using different synchronization protocols, CCI 

can be reduced and less services are required. However, further 

studies are necessary to confirm the benefit of extended VWP. 

1265 Reduced Risk of BCV and BRSV Infections in Swedish 

Organic Dairy Herds 

M. R. Bidokhti, M. Travén, N. Fall, U. Emanuelson, S. Alenius 

Swedish University of Agricultural Sciences, Division of Ruminant 

Medicine and Veterinary Epidemiology, Department of Clinical 

Science, Uppsala, Sweden 

Bovine coronavirus (BCV) and bovine respiratory syncytial virus 

(BRSV) infections are spread in cattle herds worldwide. When 

introduced in to a susceptible herd, both infections cause respiratory 

disease. BCV is also associated with diarrhea in both calves and 

adult cattle. The purpose of this study was to investigate the 

antibody prevalence of BCV and BRSV in dairy herds in an area of 

Sweden. Specific aims were to compare the prevalence of these 

infections in organic and conventional herds and to study the 

Miscellaneous 313

influence of different risk factors for seropositivity. 699 serum 

samples, taken from 20 conventional and 20 organic dairy herds in 

south eastern Sweden on two sampling occasions with one year 

interval, were tested by ELISA for presence of antibodies to BCV 

and BRSV. On individual level, the seroprevalence on both 

occasions varied between 82-86% to BCV and 79-82% to BRSV. 

Analyzing the data on herd level revealed that the conventional 

herds had a significantly higher mean seroprevalence to BCV and 

BRSV than the organic (P

32 Factors Affecting Reproductive Performance in Israeli Dairy 

Herds 

N. Galon 

Hachaklait, Caesarea Ind. Park, Israel 

Objective: The objectives of this study were to portray the 

reproductive parameters of monitored Israeli dairy herds and their 

rates. To describe Hachaklait herd-health system and to show how 

these can be used jointly to improve herd performance. 

Materials and Methods: Israel has ~100,000 Holstein cows kept on 

~1,000 highly intensive zero-grazing farms. 70% of farms are in the 

herd book and 50% of them use automatic individual cow identification 

systems. Intensive herd medicine is practiced in most herds by 

Hachaklait Veterinary Services Ltd. Regional vets visit farms weekly 

and check cows routinely several times per lactation. The cows’ data is 

collected from farm management software programs nationwide and is 

processed into a herd-health report. This report monitors for each farm 

the rates of the main calving, reproduction and milk production 

parameters. The report compares the farm’s specific performance 

profile with its previous reports and with over 100 other farms. A 

causal analysis, using logistic and linear regression multifactoial 

models, explains the effects and interactions of various risk factors 

involved on a specific farm. 

Results: Mean herd’s rates of calving traits known to affect 

reproduction in primiparous and multiparous cows respectively in 

2006 were: stillborn 7.1% and 5.9%, twins 0.8% and 6.5%, retained 

placenta 9.0% and 13.1%, endometritis 43.3% and 27.2%, ketosis 

13.8% and 17.8%. Other risk factors found to affect reproduction 

were negative energy balance, body condition score, dry period 

length, summer calving, high milk production and somatic cell 

count. The mean herd’s rates in 2006 of reproductive parameters 

commonly used in Israel were: waiting period length 99.0 and 91.7 

days, undetected heat 34.7% and 29.2%, inactive ovaries 11.0% and 

10.6%, cows not inseminated at 150 DIM 12.9% and 17.8%, cows 

open at 150 DIM 36.9% and 42.3%, days open 121.5 and 121.8 days, 

and first AI conception rate 42.6% and 33.4% in primiparous and in 

multiparous cows respectively. Different risk factors are found to 

have significant effects on different farms. 

Conclusion: Intensive veterinary service combined with detailed 

individual cow data enable the production of informative herd report, 

comparison between herds and years, and analyzing the reproductive 

efficiency of each dairy farm. 

33 Declining Fertility in Dairy Cows in Japan and Effort to 

Improve the Fertility 

T. Nakao 

Faculty of Agriculture, Yamaguchi University, Department of 

Veterinary Clinical Sciences, Yamaguchi, Japan 

Fertility of Holstein-Friesian dairy cows has been declining at a rate 

of 1 % annually since 1993 in Japan where 1.6 million dairy cattle 

are being raised and average milk yield per cow per lactation is about 

9,800 kg. First AI conception rate in lactating cows was above 54% 

until 1993, decreased to 43 % in 2003, and has still been declining 

thereafter. As a consequence, the average calving interval extended 

from 410 days to 440 days during the same period. Meanwhile, 

fertility levels in dairy heifers, beef cows and heifers after AI or ET 

have not significantly decreased. In dairy cows increase in milk yield 

and resultant negative energy balance during early lactation period 

might have caused an increased incidence of abnormal resumption of 

ovarian cycles postpartum, which may adversely affect subsequent 

fertility. Our nationwide field investigation based on milk 

progesterone measurements in 344 cows at 11 commercial dairy 

herds across Japan showed that 58.4% of the cows had abnormal 

resumption of ovarian cycles postpartum; 36.9% had delayed first 

ovulation, 10.2% had prolonged luteal phase, 11.3% had other types 

of abnormality such as cessation of the cyclicity. Abnormal ovarian 

cycles postpartum significantly decreased subsequent reproductive 

efficiency. Nutritional management of cows during transition period 

to reduce peri-parturient complications and to maximize dry matter 

intake is the basic strategy to reduce delayed first postpartum 

ovulation. Vitamin and mineral supplement is often used during this 

period for prevention of retention of fetal membranes and 

hypocalcemia. Further studies are needed to establish practical 

methods for the diagnosis and treatment of prolonged luteal phase. 

Different approach is needed to decrease prolonged luteal phase. 

Synchronization of estrus and ovulation with timed AI is widely 

applied. In conclusion nutritional management of cows to reduce 

abnormal resumption of ovarian cycles postpartum and use of novel 

reproductive technology to improve heat detection rate are keys to 

improve fertility of dairy cows in Japan. 

Key words: dairy cows, fertility, postpartum ovarian dysfunction, 

reproductive performance 

64 Reproduction of Dairy Cows in New Zealand 

N. Williamson 

Massey University, Institute of Veterinary Medicine and Animal 

Science, Palmerston North, New Zealand 

Dairy production in New Zealand differs from most places. It has 

seasonally concentrated calving patterns to use pasture efficiently and 

cows are grazed outside on pasture all year. This environment limits 

production of cows and leads to specific challenges in reproductive 

management and novel approaches to overcome them. The highly 

seasonal nature of dairy farming also produces some different measures 

of reproductive performance that apply to herds where the planned start 

of calving date and planned start of mating date are specified, such as 

submission and calving rates. Fertility in New Zealand has declined in 

association with the introduction of US Holstein genetics and this may 

be due to reduced luteal function or increased clearing of progesterone 

in dioestrus. As a result, New Zealand farmers as a whole appear to 

have limited their use of Holstein genetics. 

Key words: dairy cow, reproduction, fertility, reproductive indices, 

Holstein, anoestrus, conception rates 

65 Dairy Herd Production and Reproduction in Quebec and 

Canada 

E. Bouchard, D. Du Tremblay 

Université de Montréal, Faculté de médecine vétérinaire, 3200 

Sicotte, Saint-Hyacinthe (Quebec), Canada 

In Canada, the value of products shipped by dairy processors 

amounted to nearly 11.5 billion CAD in 2005, a figure that 

represents 15.9% of the value of all products in the food and 

beverage industry. Canadian milk producers are constantly striving 

to improve their efficiency and thus maintain the viability of their 

enterprise. It is therefore critical that veterinarians develop 

specialised expertise in all areas of bovine health that have an impact 

on the performance of their clients’ herds. In most cases, the area 

with the greatest impact is reproduction. Monitoring the overall herd 

situation and improvements in reproductive performance over time 

is an essential part of veterinary practice. This article presents the 

reproductive statistics and trends observed in Quebec, the province 

with the highest milk production of the 10 Canadian provinces. 

Key words: dairy cow, reproduction, fertility, reproductive 

performance, reproductive indices 

66 Reproduction Performances and Risk Factors in Dairy and 

Beef Herds in Belgium 

Ch. Hanzen 1 , L. Théron 1 , J. Sterkendries 1 , Y. Laurent 2 

1 University of Liege - Faculty of Veterinary Medicine, Department of 

Theriogenology, Liege, Belgium 

1 Animal Production Consulting s.a., WAVRE, Belgium 

As in many different countries, the average number of animals per 

dairy or beef farm has increased in Belgium. According to these 

changes, the work of the veterinarian has to be modified. Individual 

diagnosis is the first condition to make an herd diagnosis. In that 

purpose, it’s necessary to have. a good strategy to collect and analyse 

the data. Since several years, we develop such preventive approach 

of reproduction problems by improving a computerized tool. In 

Belgium, 60 % of cows are beef cows (Belgian Blue breed) and 40 % 

are dairy cows (mainly Holstein breed) producing on average 8.000 

liters by lactation. Both are managed in an intensive fashion. 

Typically, both breeds have a whole year-round distribution of 

calvings, the highest percentage appearing between August and 

December. Age at first calving is higher in beef (28 months) than in 

dairy (26 months) cattle. Waiting, reproduction period and number 

of days open are quite similar in both speculations. Reproduction 

Reprodiction Workshop 315

performances of primiparous cows are a little worse than 

reproduction performances of multiparous cows. Fertility is similar 

in beef and dairy cows. During the last 9 years, interval between 

birth and first calving has increased in beef cows. The calving 

interval has decreased in beef cows and increased in dairy cows. 

Fertility of beef cows has increased. No changes in fertility have 

been observed in dairy cows. Quite normally, Belgian breed is 

characterized by a huge number of caesarean section (85 %). The 

frequency of the different infertility risks factors are rather low. 

Key words: reproduction, dairy cows, beef cows, Belgium 

67 A Wide Range of Tools to Improve Reproductive 

Performances in Dairy Cows 

C. Ponsart 1 , B. Frappat 2 , P. Le Mezec 2 , S. Freret 1 , H. Seegers 3 , 

P. Paccard 2 , P. Humblot 1 

1 UNCEIA, Département R et D, Maisons-Alfort, France 

2 Institut de l’Elevage, Département R et D, PARIS, France 

3 UMR INRA-ENVN, Gestion de la Santé Animale, Nantes, France 

A research and development programme conducted by UNCEIA and 

its partners has been proposed to face the decrease in reproductive 

performances observed in high producing dairy cows. Since 2002 

several tools related either to genetics and environmental/management 

effects on fertility were developed in an attempt to stop this decrease or 

even improve female fertility. An improved model for genetic 

evaluation of fertility based on data issued from the National data basis 

(SNIG) has been implemented in 2007 by INRA, and in addition, a 

survey studied the possible use of pregnancy results to complete the 

data used for indexation. As a complement to those actions related to 

quantitative genetics, research programs aiming to define genetic 

markers of fertility susceptible to be used in Marker Assisted Selection 

(MAS) have been initiated. Different programmes were conducted for 

QTL refine mapping and to identify genes differentially expressed 

according to different phenotypes and/or environmental conditions. A 

second part was dedicated to farming practices. Several field trials led 

to identify predictors of cyclicity disorders and to describe oestrus 

detection characteristics (frequency of oestrus signs and signs used to 

call AI technicians) and insemination practices / risks factors (for 

instance type of restraint) associated with lower success of 

reproduction. Finally, tools were developed to facilitate fertility 

surveys/follow up by AI cooperatives and for teaching technicians and 

farmers and in order to adopt farming practices related to better 

reproduction results. A handbook summarizing information and 

corresponding advices in the field of bovine reproduction and dedicated 

to AI technicians and farmers has been produced. A method aiming to 

diagnose and solve reproductive problems in dairy herds and session 

for farmer education based on exchanges between participants on 

management practices were proposed. The first results originating from 

last years genetic and phenotypic evaluation tend to show positive 

trends for fertility. From preliminary observations issued from a subset 

of herds positive results in terms of reproductive performances have 

been obtained too after application of recommended measures. The 

combined use of this wide range of tools should contribute to optimize 

reproductive performances in dairy herds 

68 Reproduction in Australian Dairy Herds 

R. Rheinberger 

Highlands Veterinary Group, Mittagong, Australia 

The Australian dairy industry has faced its own crisis with the returns 

from dairying diminishing with the reduced milk prices and the drought 

increasing the cost of production. Dairying spreads from the tropical 

north to the temperate south and all points in between. 

The industry invested in a huge research project, covering 40,000 cows 

and 200 commercial dairies. The outcome of the extension project as a 

consequence of the results obtained has put a new emphasis on 

standardising fertility reporting in Australian herds. The InCalf 

extension project and the Fertility Focus Report have added a new 

benchmark to the dairy industry in Australia. 

Key words: Australian dairy industry, fertility 

110 ß-Carotene in Dairy Cattle: Metabolism in the Ovary and 

Status Assessment 

J. Schweigert 


Institute of Nutritional Science, University of Potsdam, Potsdam, 

Germany 

Feeding trials in cattle as well as in-vitro experiments point to a 

specific importance of ß-carotene (ßC) in enhancing reproductive 

performance. Based on recent results from studies using cellular 

and molecular approaches, the importance of ßC as a local precursor 

of retinoids appears most probable. A conclusion based on local 

changes in levels of retinoids as well as their specific binding 

proteins and on the recent observation that the enzyme converting 

ßC into vitamin A is expressed especially in reproductive tissues in 

a coordinated manner. Follicular fluid serves as nutritional source 

for the developing oocyte and its steroid composition is an indicator 

of follicle quality. In cattle, retinol levels increase with follicle 

maturation. This increase is associated with a higher conversion rate 

of ßC into vitamin A in follicles of good quality. This indicates the 

importance of ß-carotene as a local vitamin A precursor in dairy 

cows. Furthermore, essential binding proteins are expressed in the 

ovary structures indicating a regulated supplementation to ovary 

cells. A selective accumulation of ßC has been ascribed to a specific 

function in the corpus luteum. ßC content of the corpus luteum is 

dependent on dietary supplementation and stage of development. 

ßC is converted into vitamin A and the conversion rate is highest in 

the mid-luteal phase when progesterone synthesis is greatest. This 

again supports the importance of ßC as local vitamin A precursor as 

well in the corpus luteum. An adequate supply of ßC to target 

tissues is therefore essential for dairy cows. To obtain an efficent 

supply of the ovary with ßC, sufficiently high plasma levels are 

necessary. To initiate or to optimise ßC supplementation in cattle, 

ßC status has to be determined. Different methods are available to 

determine ßC plasma levels which are either less reliable or 

expensive and time consuming. Recently, a novel cow-side assay, 

the iCheck() and iEx() system (BioAnalyt GmbH, Germany), 

has been introduced. With this novel test system blood levels of ßC 

can easily be assessed using whole blood within a few minutes at 

cow-side achieving a comparable quality as with highly 

sophisticated time consuming and expensive laboratory method. 

111 ß-Carotene in Dairy Cattle: Endocrinological and Metabolic 

Aspects Related to Fertility in the Peripartual Period 

K. Kida 

Field Center of Animal Science and Agriculture, Obihiro University, 

Obihiro, Hokkaido, Japan 

Recently due to increase in milk yield, calving interval become 

longer that is a common problem in commercial dairy cattle. One of 

the important reasons of prolonged calving interval is increased 

negative energy balance that cause delay in ovarian resumption after 

parturition. On the other hand, cows that ovulate within 3 weeks 

after calving showed normal ovarian cycle earlier and calving to first 

AI intervals shorter as well as pregnancy rate better, than 

anovulatory cows. Therefore earlier ovulation after parturition 

guarantees better reproductive performance. Cows that ovulate 

within 3 weeks after parturition show higher IGF-1 concentration 

than anovulatory cows. This means that ovulatory cows were 

managed well in dry period. However cows that have fast increase in 

daily milk production show lower ovulation rate in spite of the same 

total milk production as ovulatory cows. This indicates that fast 

increase in milk yield deteriorate negative energy balance. During 

the peri-parturient period, metabolic status changes rapidly due to 

DMI depression, fetal growth and colostrum synthesis. At the same 

time, b-carotene concentration decreases due to secretion in 

colostrum and decreasing DMI, and shows nadir 1-2 weeks after 

calving. This change in b-carotene level is similar to change of 

energy balance during peri-parturient period. It is well-known that bcarotene 

can prevent mastitis or retained placenta and have direct 

effect on development of ovarian function. Recently we found that 

early ovulated cows show higher b-carotene concentration than 

anovulatory cows in dry period. In this research, milk production of 

anovulatory cows was the same with ovulatory cows, however, SNF 

and milk protein levels were lower and they showed anemia and 

higher GGT in early lactation period. These conditions are usually 

derived from relatively prolonged malnutrition condition and 

chronic sickness; therefore these abnormalities originate from dry 

period. Namely unhealthy condition in dry period, makes cow’s 

metabolic profile abnormal and low b-carotene concentration

simultaneously, and these delay the first ovulation after calving. In 

conclusion, to ovulate early after calving, cows should be given 

better management in dry period, such as to avoid low DMI. For 

monitoring nutritional and health status, b-carotene is a good 

indicator. 

112 ß-Carotene Deficiency in Dairy Cattle: Reasons and 

Prevention 

W. Schliffka 

Dairy Production Research Centre, Germany 

An adequate supply of ß-Carotene is of prime importance for dairy 

cows. ß-Carotene is not only the natural precursor of vitamin A for 

ruminant animals; it has also a very important function as a fertility 

vitamin in these species. Access to pasture or good quality grass 

silage should guarantee an adequate supply of ß-Carotene to dairy 

cows. However, modern rations for high yielding dairy cows are 

often based on total mixed rations (TMR). TMRs in early lactation 

frequently contain more than 50% maize silage and cows usually 

have little or no access to pasture. Maize silage has a negligible ß- 

Carotene concentration. The capacity of grass products to 

compensate for the low ß-Carotene content in maize based rations is 

often overestimated. DSM survey results from 1999-2007 show that 

the ß-Carotene content of grass silage is highly variable and 

unpredictable. This is very much related to the crop and conservation 

technique. Moreover, the oxidative losses of ß-Carotene during 

harvest, wilting and ensiling can be considerable. Compared to the 

pre-ensiled fresh grass, ß-Carotene losses can easily exceed 50% 

during storage and decreases further once the silo is opened. The 

correlation between the ß-Carotene concentration of the diet and the 

ß-Carotene concentration in plasma of dairy cows is very strong. The 

optimum level of ß-Carotene in the blood should exceed 3.5 µg/ml. 

Recent surveys of blood plasma samples in intensive dairy herds in 

selected countries worldwide showed that the majority of the cows 

do not exceed ß-Carotene levels of 2.5 µg/ml plasma. These findings 

confirm that the ß-Carotene concentration in dairy cow rations is 

often insufficient to fulfill the requirements in early lactation. An 

inadequate supply of ß-Carotene leads to a dramatic decline in 

reproductive performance. The following are typical symptoms, 

weak or silent heat, delayed ovulation, low fertility rate, increased 

occurrence of cystic ovaries, early embryonic losses and retained 

placenta. Reproductive disorders result in losses of up to 150 _ per 

cow per year. They play a key role for early culling in dairy herds. 

Inadequate dietary ß-Carotene can be easily rectified with 

supplementary ß-Carotene added to minerals or compound feed. If 

blood levels are < 1.5 µg/ml the cow is deficient and 

supplementation of 500 mg ß-Carotene/cow/day is recommended. 

Levels between 1.5 – 3.5 µg/ml are classified marginal and require at 

least the supplementation of 300 mg ß-Carotene/cow/day. The 

supplementation should start at the beginning of the dry period to 

support ovum development and heat intensity but also for optimum 

fortification of the colostrum. Supplementation should be continued 

until the next pregnancy is confirmed. 

113 Factors Related to the Reproductive Performance of Dairy 

Herds in Chile 

L. Poo 

Private, Clínica de Rumiantes, Osorno, Chile 

Chile is a very long country and the climate is very different between 

north and south. The main milk production areas are in the south with a 

production of more than 70% of the total country milk production. 

Basically there are two models of milk production. One is based on the 

use of cows with high volume of individual productions which are 

frequently fed with high amount of concentrated feed. In Chile several 

aspects including management, nutrition and genetics are responsible 

for the reproductive level in cows and this topic is discussed. 

Reproductive parameters more frequently used in Chile are discussed 

and the conclusion is that the reproductive performance of dairy herds 

in the southern Chile is not affected by serious problems but it must be 

improved in factors such as heat detection, age at first calving, 

nutrition, artificial insemination use, as well as record keeping and use 

of ultrasonographic techniques. 

Key words: Pregnancy rate, conception rate, heat detection, open days, 

voluntary waiting period 

114 Factors Responsible for Increased Calving Intervals in Crossbred 

Cows in India 

A. Nanda, J. Singh 

Guru Angad Dev Veterinary & Animal Sciences, Animal 

Reproduction, Gynaecology & Obstetrics, Ludhiana, India 

India made tremendous progress in dairy production through 

crossbreeding of indigenous low yielding cows with high yielder 

exotic breeds, especially HF, over the last two decades. Compared to 

indigenous cows, the crossbreds mature early and calve first at the 

age of 31.1 vs 44±2.41 months, and have shorter Inter Calving 

Intervals (ICI; 437±28.9 vs 517±53.8 days). However, the crossbred 

generations from F2 and beyond have increased reproductive 

problems and their ICI increased to 451±34.3 days or even longer. 

Conditions like repeat breeding syndrome (RBS; 20-39%), anestrus 

(14-48%) and cystic ovaries (30%) have been increasing. Cows with 

>75% exotic blood have lower uterine immunity than natives and 

hence higher incidence of endometritis. Insufficient pre- and 

postpartum nutrition and unhygienic, unscientific conventional 

animal husbandry practices also favored uterine infections. Stress of 

high milk production, seasonal extremes, and lameness, add to 

hormonal abrasions and infertility. Modulation of uterine immunity 

in crossbred cows with microbial endometritis has been proposed as 

an alternate to conventional treatment with antibiotics / antiseptics 

etc. Single intrauterine infusion of 100 g E. coli lipopolysaccharide 

(LPS) in 30 ml normal saline or of 500 mg Oyster Glycogen in 50 ml 

normal saline induced an influx of neutrophils into the uterine lumen 

within 6h. These cleared uterine infections in more than 75% repeat 

breeding endometritic cows through enhanced phagocytosis and 

substantially improved their fertility (CR 45 - 66.6% vs 12.6% in 

untreated controls). General immunomodulation of such cows 

through parenteral administration of 10 ml Inmodulen once or twice 

(25 mg inactivated cells of Propionibacterium granulosum and 2.0 

mg E. coli LPS per 100 ml) also been proposed. Management of 

fertility in repeat breeding cows with prolonged estrus, estrus with 

suprabasal circulatory P4 and luteal insufficiency could be better 

made through either (i) stimulating LH release with exogenous 

GnRH or (ii) by early termination of luteal phase using 

PGF2·. Although good quality fodder/balanced feeding is 

indispensable, supplementation with urea molasses multinutrient 

blocks feeding or parenteral soybean oil emulsions proved beneficial 

in improving fertility and attaining normal ICI. A comprehensive 

approach, however, would need to be involved to deal with infertility 

in a holistic way in crossbred cows in India. 

Key words: Reproduction, Crossbred Dairy Cow, India 

115 Prevalence, Causes, Treatment and Prophylaxis of Cow 

Infertility in Poland 

E. Malinowski 

National Veterinary Research Institute, Department of 

Pathophysiology of Reproduction and Mammary Gland, Pulawy, 


The efficiency of reproduction is the best indicator of the adequate 

management, feeding, housing, welfare and good health status of 

dairy cows. Diseases of the genital tract, metabolic disorders, 

mastitis, and lameness are the main causes of subfertility or 

infertility in cows. Incidences of the genital tract diseases 

significantly ranged among different herds. Embryo mortality and 

abortions were noted from 0 to 11.4%, dystocia from 1.2 to 19.4%, 

retained fetal membranes from 3.4 to 31.2%, puerperal metritis from 

0 to 40.6%, clinical endometritis from 2.8 to 34.7%, ovarian cysts 

from 0 to 50%, ovarian afunction from 1.4 to 50% and silent heat 

from 1.6 to 49% of cows. Other diseases such as clinical mastitis 

ranged from 13 to 72%, clinical ketosis (0-14.5%), lameness (0- 

45.3%) and displaced abomasum from 0 to 4% of cows in particular 

herds. Fertility indicators: conception rate (26-63%), artificial 

insemination index (1.6-3.5), calving-pregnancy period (88-178 

days), and culling rate (2.1-12.5%) also differed among the 

examined herds. A deficiency or excess of energy and protein, and 

deficiency of antioxidants were the main feeding factors that 

negatively affected the fertility. Reproductive disorders were also 

connected with BVD-MD and IBR-IPV. Infections of the uterus 

were caused by Arc. pyogenes, E. coli and other coliforms, 

Streptococcus spp. Staphylococcus spp., Bacterioides spp. and other 

Reprodiction Workshop 317

anaerobes. Aerobic bacteria isolated from clinical 

metritis/endometritis cases were highly susceptible to 

amoxicillin/clavulanic acid. In treatment of infertility in cows, 

hormones and antibiotic products are mostly applied. The usefulness 

for treatment and prophylaxis of antibacterial compounds, 

hormones, antioxidants or immunomodulators and combinations of 

this medicine was tested by different scientific teams. Apart from the 

influence of the genital tract diseases, metabolic disorders and 

mastitis on the reproduction efficiency in cows, examinations were 

also conducted that concerned the regulation of the corpus luteum 

function, effects of xenobiotics on the genital tract, molecular basis 

and maternal recognition of the pregnancy and early embryo 

mortality. 

Key words: cow, infertility, diseases, treatment, prophylaxis 

116 Factors of Non-infectious Nature Affecting Early Foetal Loss 

in High Producing Dairy Herds in North-Eastern Spain 

F. López-Gatius 1 , O. Szenci 2 , G. Bech-Sábat 1 , I. García-Ispierto 3 , 

B. Serranod 4 , P. Santolaria 5 , J. Yániz 5 

1 

University of Lleida, Department of Animal Production, Lleida, 

Spain 

2 

Szent István University Faculty of Veterinary Science, Department 

of Animal Production, Budapest, Hungary 

3 

Autonomous University of Barcelona, Anatomy and Embryology, 

Barcelona, Spain 

4 

C.I.T.A., Gobierno de Aragón, Department of Animal Production, 

Zaragoza, Spain 

5 

University of Zaragoza, Department of Animal Production, Huesca, 

Spain 

Following a positive pregnancy diagnosis, early foetal loss is 

becoming the commonest complication of gestation in high 

producing dairy herds in North-Eastern Spain. We present here data 

derived from 14 studies and one abstract published in peer-reviewed 

journals during the period 1996 to 2008 that analysed the effects of 

factors of a non-infectious nature affecting early foetal loss in high 

producing dairy herds. Results included a total of 15,525 

pregnancies in well-managed, commercial, Holstein-Friesian high 

producing dairy herds over the period 1987 to 2007. Since the effects 

of different factors practically did not vary throughout the years, data 

from the different studies were pooled and the relative risk of every 

individual factor on foetal loss is described. Points to be highlighted 

should include that strong risk factors for pregnancy loss were parity 

(lactating cows versus heifers), semen-providing bull, warm season 

and twin pregnancies, whereas the presence of an additional corpus 

luteum was identified as a strong positive factor favouring the 

maintenance of gestation. Progesterone supplementation had the 

potential to reduce the incidence of pregnancy loss. From a practical 

point of view, assessment of normal development of gestation on 

days 60 and 90 after insemination is suggested. 

Key words: early foetal loss, milk production, dairy cows 

117 Performance Parameters and the Predicted Economic Return 

of Jersey Holstein-Friesian Crossbred Cows in Australian 

Pasture-based Herds 

M. Pyman, K. Macmillan, G. Anderson 

University of Melbourne, Department of Veterinary Sciences, 

Werribee, Australia 

This study compared the reproductive performance and milk 

production of Holstein-Friesian cows to that of Jersey Holstein- 

Friesian crossbred cows in 14 Australian herds. An economic model 

using the study results was developed to determine the profitability of 

crossbreeding in pasture-based, seasonally calving herds. Data were 

collected over a single lactation from 3,347 cows comprising 2,373 

Holstein-Friesian (FFFF), 669 Jersey Holstein-Friesian first cross 

(JJFF) and 305 backcross (FFJF) cows. First service conception rates to 

artificial insemination were based on 2,179 FFFF and 894 crossbred 

cows. Reproductive performance of the crossbred cows was superior 

to the FFFF cows for first service conception rate (54.7% vs. 47.0%, 

p=0.0002), pregnancy rate after 14 weeks of breeding (83.7% vs. 

77.0%, p

measures of fertility in German dairy herds. Raw milk production 

and composition, reproductive events and lactation number (LN) 

data from nearly 40,000 individual cow lactations were collected 

from the Verden and Bavaria Calculation Centers of German 

Breeding Test Organizations. A total of 28,620 completed lactations 

with all required data were used for analysis. Individual cow 

reproductive performance was characterized by time to first service 

(TFS), days open (DO), calving interval (CI), services per 

conception (SPC) and conception risk (CR). Parameters for 

lactational performance included milk per day (MPD), lactation 

yield (MY), milk fat (MF), milk protein (MP) and calculated milk 

fat-to-protein ratio (FPR). Milk parameters were summarized by 

days in milk groups of ≤40 (MPD40, MF40, FPR40), 41-to-80 

(MPD80, MF80, FPR80) and >105 (MPD105, MF105, FPR105). 

Production parameters were stratified by quartiles for analysis. 

Interrelationships between production and reproductive parameters 

were analyzed using ANOVA and logistic regression models. Higher 

milk yield, irrespective of lactation stage, resulted in longer (P

P4 12 to 24 hours before calving is the best and most objective 

indication of parturition. A validated progesterone rapid blood test 

(PRBT) was applied to support the findings of the clinical examination 

in indifferent cases. This test detects an active Corpus luteum during 

cycling and is validated for cows. 

Material and Methods: A parturition score (PS) was established using 

external obstetric examination. Parameters considered were: relaxation 

of the broad pelvic ligaments, relaxation of the tail, filling of the teats 

and oedema of the vulva. A purchasable PRBT was used in cases the 

clinical score gave no clear result. 

Results: An association was found between the parturition score and 

the time of parturition. Two cut points for the parturition score were 

identified. We could exclude parturition within the next 24h in all cases 

if the animals were below the upper cut point. Animals with values 

above the upper cut point were found to deliver their calf within 24h in 

all cases. If the score was between the two cut points we deployed the 

PRBT as an additional mean to predict calving. The results show a 

sensitivity of 89.42% and a specificity of 96.15% of the PRBT to 

identify the drop of P4 before calving. The positive predictive value of 

this test is found to be 73.53% and the negative predictive value 98.7%. 

Conclusions: We have established a validated method to predict the 

time of parturition under field conditions. With this new tool 

veterinarians are able to predict the day of parturition and thus have 

better options to improve the health of the cows and/or the calf. 

The detailed composition of the score, its application and the results 

will be presented. 

Key words: calving prediction; progesterone 

154 Prediction of Calving on the Basis of Cattle's Precolostrum 

G. Pohn 1 , O. Szenci 2 , Zs. Csapó-Kiss 1 , Zs. Mándoki 1 , L. Faragó 1 , 

M. Tesszler 1 , J. Csapó 1 

1 

University of Kaposvár, Faculty of Animal Science, Kaposvár, 

Hungary 

2 

Szent István University Faculty of Veterinary Science, Clinic for 

Large Animals, Üllô, Hungary 

In the last 3 years we determined the composition of the precolostrum 

from the day 20 before calving of 55 Holstein-Friesian cattle for drymatter, 

ash content, potassium, sodium, calcium, phosphorus, zinc, iron, 

copper and manganese content, amino acid composition and fatty acid 

composition of the precolostrum. Dry-matter content of the precolostrum 

was around 35% on the days 8-12 before calving, and decreased till the 

day of calving to 25-28%, crude ash content increased with 0.6% on the 

day 12 before calving to 1.2% measured on the day of calving. Calcium 

content increased from 500 mg/kg to 1,500 mg/kg, while phosphorus 

content from 250 mg/kg to 1,400 mg/kg till the day of calving. Sodium 

content varied between 1,400 and 1,800 mg/kg, and then decreased to 

1,100 mg/kg on the day of calving. Potassium content increased during 

day 2 after calving to 1,000 mg/kg, magnesium content varied between 

180 and 220 mg/kg, with its maximum on the day of calving with 230 

mg/kg. Copper content decreased from 0.27 mg/kg measured on day 12 

to calving to 0.10 mg/kg directly at calving, whereas copper hardly 

showed changes in the function of time before calving. Changes in the 

zinc, iron and manganese content of the precolostrum before calving 

show very large dispersion, however the tendency of the change is not 

clear. With maximum reliability, sodium and potassium can ensure the 

forecast of calving; so when their concentrations show nearly identical 

values it indicates a calving occurring soon. The joint increase of calcium 

and magnesium can also provide information on the forecast of calving 

as both macro elements reach the highest value on the day of calving. 

Comparing the test stripes, measuring the macro elements, with the 

results of the atomic absorption measurements, it can be assessed that 

their sensitivity is not sufficient to monitor the changes in the minerals of 

the precolostrum. Consequently, test stripes are unsuitable for the 

forecast of the date of calving. Examining the amino acids and fatty 

acids, we could not find any change that could inform us about the 

expected date of calving. In conclusion, none of the measured parameters 

can clearly indicate the expected date of calving. It appears that it is not 

possible to forecast calving with the use of the quick tests which are used 

on farms, with the required reliability. 

This study was supported by OMFB 173/177/179-2006 

155 Parturition Induction in Dairy Cattle to Decrease Losses in 

Reproductive Performance due to Potentially Large Calves 

A. Villarroel 1 , V. Lane 2 


1 


United States 

2 

University of California, Population Health & Reproduction, Davis, 

United States 

Objective of study: The objective of this study was to determine 

whether parturition induction in dairy cattle past average due date 

would decrease the risk of stillbirth without increasing the risk of 

retained placenta. 

Materials and Methods: Dairy cattle on a single farm that reached 282 

days of gestation were administered a single dose of 0.1 mg/kg of 

dexamethasone IM. Data on the incidence of stillbirths, retained 

placenta and milk production were compared to cows that reached 282 

days of gestation and were not induced during the year following the 

study. 

Results: There was no difference in the incidence of stillbirths and 

dystocia in induced and non-induced cows. Incidence of retained 

placenta tended to be lower in induced cows than non-induced cows, 

although small sample size precluded from statistical significance. 

There was no apparent difference in milk production between induced 

and non-induced cows. Reproduction in the following lactation was 

better in induced cows than non-induced cows. Culling due to 

reproductive reasons was significantly lower in induced cows 

compared to non-induced cows. 

Conclusions: Our results show that parturition induction in cows past 

due date can increase herd profitability when compared to allowing 

cows an extended gestation period. Induction should be considered as a 

management tool in dairies to decrease future reproductive problems in 

cows that could have an extended gestation period. 

Key words: parturition induction, stillbirths, retained placenta 

156 Calf Mortality in Norwegian Dairy Herds 

SM. Gulliksen 1,2 , KI. Lie 3 , O. Osteras 2 

1 

TINE Norwegian Dairies, Dairy Production, Aas, Norway 

2 

Norwegian School of Veterinary Science, Department of Production 

Animal Clinical Sciences, Oslo, Norway 

3 

Norwegian School of Veterinary Science, Department of Pathology, 

Oslo, Norway 

The aims of the present study were to estimate the mortality rate in 

calves and young stock up to 365 days of age in Norwegian dairy 

herds, and to identify associations between calf mortality and 

possible risk factors on both calf- and herd level. Approximately 90 

% of all Norwegian dairy farmers are registered members of the 

Norwegian Dairy Herd Recording System (NDHRS). In the current 

study, information on 341.712 calvings in 15.050 Norwegian dairy 

herds in 2005 was extracted from this system. The overall calf 

mortality rate during the first 6 months of life, in herds registered in 

the NDHRS was found to be 6.3 %, including abortion and stillbirth. 

First parity cows were found to have lower risk of abortions, but 

higher risk of stillbirth compared to older cows. Calvings during 

autumn months (September - November) were found to be beneficial 

both in respect to the risk of abortion and stillbirth, while calves born 

between June and October was found to have the lowest risk of death 

during the first month of life. Dystocia and occurrence of congenital 

malformations were found to be by far the most important factors 

influencing the risk of stillbirth and death during the first week, 

describing 14.4 and 1.68 % of the variation, respectively. 

Occurrence of arthritis was the most important disease influencing 

the risk of death in calves during the first month of life, although 

diarrhoea and respiratory disease were the most frequently reported 

diseases in this age group. Respiratory disease was found to be the 

most important disease influencing the risk of death in calves 

between 31 and 180 days of age, as well as for calves of more than 

180 days of age. The calf mortality rates in all age groups were found 

to increase with increasing herd size. Besides herd size, the 

occurrence of respiratory disease, arthritis or gastritis or enteritis 

were the only significant factors found to influence the risk of death 

in calves older than 180 days of age. A significant cluster effect 

within herd was found in all age groups. As dairy cattle farming in 

Norway moves towards increasing herd sizes and new housing 

systems, subsequent new challenges when it comes to calf 

management must be prioritized to avoid increasing morbidity and 

mortality rates. 

157 Stillbirth in Heifers: Experiences in the Netherlands

J. Muskens 

Animal Health Service Deventer, Deventer, Netherlands 

Stillbirth in heifers was an important issue for many Dutch dairy 

farmers during the last two decades. A pilot study of 10 dairy herds 

with at least 20% of stillbirth of heifers was performed. The study 

consisted of extensive post-mortem examinations of one stillborn calf 

per herd, blood and urine testing of pregnant young stock at 0.5-2 

months before expected calving, and a questionnaire about parturition, 

nutrition and management around calving. Nine herds were certified 

leptospirose free; one herd was treated with antibiotics. One calf was 

BVDV positive at post mortem examination. In none of the calves there 

were indications of infections with Neospora caninum or Salmonella 

spp., or of a deficiency of iodine. The number of herds with decreased 

202 Mycoplasma Diseases in the Bovine: Diagnosis, Prevention 

and More 

L. Fox 1 , C. Schneider 2 , L. Stipkovich 3 

1 Washington State University, Veterinary Clinical Sciences, Pullman, 

United States 

2 University of Idaho, Moscow, ID, Animal & Veterinary Sciences, 

Moscow, United States 

3 Veterinary Medical Research Institute of the Hungarian Academy of 

Sciences, Veterinary Medicine Research Institute, Budapest, 

Hungary 

The Mycoplamsa sp. that cause mastitis are simple, cell wall-less 

bacteria that can colonize and cause diseases in other extramammary 

sites in the bovine. Prevalence of mycoplasma mastitis 

appears to be increasing in many locations throughout the world. 

Traditionally Mycoplasma sp. mastitis has been considered 

contagious pathogens that are transmitted in a fashion that is typical 

of other contagious mastitis pathogens (S. aureus and Str. 

agalactiae). Such transmission is believed to occur mostly during 

milking time, and that introduction of mycoplasma mastitis can 

occur through the importation of infected cattle from outside the 

dairy. Studies will be reviewed that have indicated that transmission 

of Mycoplasma sp. can occur internally. From the extra-mammary 

nidus of colonization, Mycoplasma sp. can be transmitted to the 

mammary gland, or vice versa. New prevention methods would have 

to be developed to counter this mode of transmission. The 

established contagious mastitis control practices of employing strict 

milking time hygiene that includes post-milking teat asepsis, and of 

identification of the infected animals and isolation of those animals 

by separation and culling, has been successful and is currently the 

method advocated to control mycoplasma mastitis outbreaks. The 

fault in this system is that although it controls the outbreaks, it might 

not be as effective at preventing the outbreaks from occurring. In 

addition to mastitis, Mycoplasma sp. bacteria have been implicated 

in a variety of disease conditions in cattle including 

bronchopneumonia, septic arthritis, reproductive disorders and otitis 

media. The overall annual impact of this etiologic agent on U.S beef 

and dairy industries has been estimated to be as high as $100 million. 

Calf hood infections may play a role in the transmission of 

Mycoplasma sp. in particular, more virulent species such as 

Mycoplasma bovis on dairies. Basic mechanistic and applied 

Mycoplasma sp. research holds promise for elucidating the 

underlying etiology and risk factors associated with calf hood ear 

infections. Related areas of research in the U. S. concern the role of 

Mycoplasma ovipneumoniae in acute, severe, fatal outbreaks of 

respiratory disease in lambs occurring in indigenous native sheep 

populations (Ovis canadensis). Emerging evidence indicates that 

Mycoplasma sp. may be acting as primary pathogens in these disease 

outbreaks. 

203 Mycoplasma Bovis Infection of Calves 

L. Stipkovits 1 , S. Szathmary 2 

Mycoplasma Workshop 

blood contents of GSH-Px or copper, or decreased urine contents of 

magnesium in young stock was 2, 3 and 2. Five herds had at least one 

animal with GSH-Px contents higher than 600 U/g Hb. Five farmers 

reported that the afterbirth was delivered within one hour of calving; 

four farmers reported that most heifers had retention of the afterbirth. 

Five farmers reported an increased mean body condition score of the 

pregnant young stock. In 2006 187 stillborn calves were examined post 

mortem at the Animal Health Service in Deventer. Of these, 87 calves 

had no abnormalities. Signs of asphyxia were diagnosed in 73 calves. 

Other diagnoses included congenital defects (n = 8), infections with 

BVDV (n = 8) or Arcanobacterium pyogenes (n = 4). Based on these 

results, other Dutch field experiences and literature, a possible 

approach to dairy farms with an increased numbers of stillborn calves 

in heifers will be discussed. 

1 Vet.Med.Res.Inst., Mycoplasma, Budapest, Hungary 

2 Galen Bio Inc., Mycoplasma, Carlsbad, United States 

In cattle, beside Mycoplasma mycoides subspecies mycoides SC, 

being present in African and Asian countries, M. bovis is the most 

pathogenic species. The infection can be introduced in previously 

negative herd by clinically healthy infected animals, by various tools 

used in barns, by milk from infected cows and by semen. If the 

infected calves are introduced in the groups of negative calves 

clinical symptoms of respiratory diseases start to develop. Serouspurulent 

nasal discharge can be observed after 4-7 days, increased 

rectal temperature, frequent cough, difficulty in breathing and 

lethargy develops later after mixing the animals. Over 20% of the 

animals in the group can develop mono- or polyarthritis, or otitis, or 

meningitis. Weight gain of the diseased calves is decreased 

significantly. Mortality due to pneumonia in such infected herd, if 

treatment is not performed, can reach 25-30% of calves during first 

40 days. Later on animals became asymptomatic, but they carry M. 

bovis and can be a source of infection for the other animals. These 

asymptomatic animals will still show delayed growth. Body weight 

at age 8-10 month is about 100 kg less than the uninfected 

animals. Infection with M. bovis through semen results in abortion, 

early birth of weak calves, with signs of pneumonia and arthritis. 

This can be recorded in few days old calves. About 50% of the 

infected cows have retention of embryonic membranes after calving 

and develop parenchymatosus mastitis with very high increase in 

somatic cell count along with significant decrease in milk 

production. Some of the cows also show arthritis. If the milk from 

cows suffering from M. bovis mastitis is fed young calves, 

respiratory symptoms develop in 3-6 days. If the infection is 

introduced through healthy-looking, but infected heifers or cows, 

symptoms of the disease (respiratory disease, abortion or mastitis) 

develops gradually in relatively fewer animals. Later on the disease 

spreads more rapidly. Post mortem examination of young calves died 

due to M. bovis infection, inflammation of the trachea, pleuritis and 

catarrhal pneumonia of various extensions, enlarged peribronchial 

lymph nodes, sometimes inflammation of kidney can be observed. In 

the arthritic joint, accumulation of serous-fibrinous exudates, 

erosion of the synovial membrane can be detected. Mycoplasmas can 

be cultured from several organs. 

Key words: cattle, Mycoplasma bovis, isolation, pathology 

204 Prevalence of Mycoplasma bovis Infection in French Dairy 

Cattle 

MA. Arcangioli 1 , MA. Botrel 2 , M. Chazel 2 , E. Sellal 3 , P. Bézille 1 , 

F. Poumarat 2 , D. Le Grand 1 

1 

Ecole Vétérinaire, Ruminant Mycoplasmosis, Marcy l'Etoile, Lyon, 

France 

2 

AFSSA, Ruminant Mycoplasmosis, Lyon, France 

3 

LSI, Diagnostic, Lissieu, France 

Mycoplasma bovis (Mb) is widely described as a causal agent of 

pneumonia in calves and mastitis in dairy cows. Very little information 

Mycoplasma Workshop 321

exists about Mb prevalence in dairy herds in France. A national 

serological survey previously showed that in French beef cattle an 

average of 7% of animals and 28 to 90% of the herds, according to the 

department, have been infected by Mb. Two recent studies were 

conducted to try to determine the prevalence of Mb udder infection in 

dairy cattle. The first one was implemented with the aim of estimating 

the Mb prevalence in the 1522 herds of a milk cooperative covering six 

southeastern French departments. The number of herds to be tested 

(n=345) was calculated based on an expected Mb infection prevalence 

rate of 2% (by reference to US data). The second one was conducted 

over a period of 12 months, based on 280 dairy herds coming from the 

same region, in order to estimate the prevalence of Mb in clinical 

mastitis. Each month, only one clinical mastitis milk sample was 

allowed to be collected from the different herds. Mb search was carried 

out by i) culture and then identification by dot immunobinding and ii) 

PCR directly performed on samples (kit LSI-Lissieu-France). 

Occurrence for M. canadense, M. bovigenitalium and M. alkalescens 

was also tested. The results demonstrated that the prevalence for Mb 

infection in these dairy herds was lower than the 2% expected. Both 

surveys were preliminary studies and will need to be supported by 

further investigations in order to confirm our results in other areas of 

France with different dairy farming systems. 

Key words: Mycoplasma bovis, milk, dairy herds, prevalence 

205 Epidemiological Surveillance of Ruminant Mycoplamosis in 

France 

F. Poumarat 1 , MA. Arcangioli 2 , M. Chazel 1 , D. Le Grand 2 , 

P. Gaurivaud 1 , F. Tardy 1 , D. Calavas 1 

1 AFSSA, Ruminant Mycoplasmosis, Lyon, France 

2 Ecole Vétérinaire, Ruminant Mycoplasmosis, Marcy l'Etoile, Lyon, 

France 

A network of diagnosis laboratories coordinated by the French Food 

Safety Agency (AFSSA) allows a continuous epidemiological 

surveillance of ruminant mycoplasmosis throughout French territory. 

The aims are: i) to identify mycoplasmas isolated from the ruminants; 

ii) to follow the evolution of these mycoplasmosis and to detect 

emergence of new species or variants; iii) to identify any re-emergence 

of Contagious Bovine Peripneumoniae (CCBP); iv) to share scientific 

information and technical knowledge relative to the ruminant 

mycoplasmosis; v) to collect and keep representative collection of 

isolates. Mycoplasmas are first isolated from field samples in 

veterinary diagnostic laboratories and then send to AFSSA Lyon. 

Species identification is performed by dot immunobinding assay, and 

PCR is used to confirm if necessary. Then, for unidentified strains, 

conclusion is obtained by sequencing of 16S RNA genes and 

comparison to data bases. Between 2003 and 2007, about 2000 

mycoplasmas isolates from animals originating from more than 70 out 

208 Prevalence of Salmonella as Cause of Abortions in Cattle in 

Southwest Ireland and Protective Effect of Vaccinations 

J. Crilly 1 , E. Power 2 , C. Sanchez-Miguel 2 , D. Dufe 3 , B. Makoschey 4 

1 Dairy Production Research Centre, Cork, Ireland 

2 Regional Veterinary Laboratory, Cork, Ireland 

3 Intervet-Schering-Plough, Regulatory Affairs, Boxmeer, Netherlands 


Objectives: The purpose of this study was to estimate the prevalence 

rates of Salmonella Dublin (S. Dublin) as the cause of bovine abortions 

in southwest Ireland and to quantify the protective effects of S. dublin 

vaccination under field conditions. 

Materials and Methods: Pre-existing data on submissions from 

aborted and stillborn calves to the Cork Regional Veterinary 

Laboratory from 1989 to 2006 were used in the analysis. The 

material was tested for Salmonella spp by standard bacteriological 

culture. By linking the vaccination history of individual submissions 

to the post-mortem bacteriological findings, it was possible to assess 


Abortions Workshop 

of 96 French departments were identified. 1100 from cattle were 

analysed over the period. 95% came from bronchopneumonia affecting 

mainly young animals (87%) and 5% from arthritis and mastitis. 

Mycoplasma (M.) bovis was the most commonly identified pathogen 

(43%), singly or in mixed culture (20%). Otherwise M. bovirhinis and 

M. arginini were regularly found respectively (23% and 13%) but are 

thought to be only opportunistic. M. bovigenitalium, M. canadense, A. 

laidlawii and M. mycoides subsp. mycoides biotype Large Colony were 

sporadically encountered. Few isolates of M. canis and M. alkalescens 

that has been suggested to be cause of respiratory disease, arthritis and 

mastitis in cattle have been isolated but these mycoplasmas has not 

shown in France the epizootic increasing that has been reported 

recently in UK. No strain of M. mycoides subsp. mycoides biotype 

Small Colony, the agent of CBPP, was detected either in cattle or in 

small ruminants. These results underscore the importance in France of 

M. bovis in cattle pneumonia. But M. bovis mastitis incidence seems to 

be low as confirmed by other targeted surveys, only 10 outbreaks have 

been identified for 10 years. 

Key words: ruminants, mycoplasma, epidemiology, network, 

surveillance 

206 Prevalence and Control of Mycoplasma bovis in Europe 

R. Nicholas 1 , E. Raedelli 2 , M. Luini 3 , G. Loria 4 , R. Ayling 1 , 

E. Scanziani 2 

1 Veterinary Laboratories Agency, SEB, Addlestone, United Kingdom 

2 University of Milan, Milan, Italy 

3 Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia 

Romagna Brescia, Milan, Italy 



6 University of Milan, Milan, Italy 

Mycoplasma bovis is a major cause of disease in both growing and 

adult cattle. Pneumonia, mastitis, arthritis and otitis have all been 

associated with mycoplasma infection, none of which respond well 

to chemotherapy. In the UK serological screening of pneumonic 

herds indicates that M. bovis is associated with approximately a 

quarter of all outbreaks in calves; this mycoplasma is also the most 

commonly isolated pathogen from pneumonic lungs. Since its first 

introduction into Ireland in the mid 1990s, M. bovis has also become 

a frequent isolate from pneumonic lungs. In a recent study of cattle 

rearing regions in Northern Italy, M. bovis was the most common 

bacterial pathogen involved in bovine pneumonia; all veal calves and 

three quarters of all beef cattle were seropositive to M. bovis. A 

commercial vaccine still appears a long way from availability in 

Europe but autogenous vaccines, which are being used increasingly, 

may help to bridge this gap in the short term. 

Key words: Mycoplasma bovis, pneumonia, autogenous vaccine 

the benefit of S. dublin vaccinations under field conditions. All 

vaccination were performed with inactivated S. Dublin vaccines 

(Intervet International, The Netherlands). More than 16,000 

submissions from cases of bovine abortions were made over the 18 

year study period. 

Results: Apart from a small number of cases, which were unsuitable 

for bacteriological examination due to advanced autolysis, culture 

results were available for S. Dublin for most of the submissions. Of 

all samples examined for S. Dublin, 9.9% were culture positive. 

About 25% of the submissions were from cows vaccinated against S. 

Dublin, 25% cows not vaccinated against this pathogen and 50% 

with unknown vaccination status. The level of vaccination against S. 

Dublin clearly increased during the study period. The time series of 

abortions caused by S. Dublin showed evidence of two components, 

namely a background prevalence with periods where levels increase 

two fold or more. 

Conclusions: In submissions from vaccinated cows, the relative risk of 

being S. Dublin culture positive was consistently lower than for 

submissions from unvaccinated animals or animals with unknown

vaccination status (average 1989-2007: 7% for vaccinated animals 

versus 12% for unvaccinated animals). The benefit of salmonella 

vaccination appeared greater from 1996 onwards. The relative risk of 

being S. Dublin culture positive was only 5% for submissions from 

vaccinated animals after 1995 compared to 13% for submissions from 

unvaccinated animals in the same period. 

209 Sero-Epidemiology of Brucellosis in Goats at Rakh Khaire 

Wala Farm District Layyah, Pakistan 

M. Arshad 1 , M. Munir 1 , Z. Abbas 2 

1 

University of Agriculture, Department of Microbiology, Faisalabad- 

38040,, Pakistan 

2 

University of Agricuture, Department of Parasitology, Faisalabad, 

Pakistan 

Introduction: Brucellosis is responsible for considerable economic 

losses due to abortion, low milk production and infertility in goats. It 

also has a zoonotic importance that plays a significant role in the 

national economy and the public health of many parts of the world 

particularly in developing countries. Keeping in view the economic and 

zoonotic importance of this disease, the project had been planned to 

study the sero-prevalence of brucellosis in goats at Rakh Khaire Wala 

Farm, District Layyah, Pakistan. 

Materials And Methods: A total of five hundred (500) serum 

samples were collected randomly from goat breeds (Teddy, Pak 

239 The Present and the Future of IBR Control 

P. Franken 

Animal Health Service Deventer, Cattle Health Service, Deventer, 


The development of a marker vaccine against IBR, has opened the 

way for the eradication of IBR also in high prevalence and high 

density area's. A number of European countries have started or will 

start eradication programmes based on the use of this vaccine. 

Actually there is a lot of knowledge and experience about control 

and eradication of IBR in different epidemiological situations.An 

overview will be given of the present status in Europe and the needs 

for international trade of animals and genetic material will be stated. 

Key words: IBR, marker vaccine, control, future 

240 Infectious Bovine Rhinotracheitis and the Epidemiological 

Role of the other Ruminant Species 

J. Thiry, B. Muylkens, E. Thiry 

University of Liege - Faculty of Veterinary Medicine, Department of 

Infectious and Parasitic Diseases - Virology and Viral Diseases, 

Liege, Belgium 

See in the Supplement I. 

241 Pitfalls in IBR Diagnosis in the Field 

M. Beer, P. Koenig 

Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, 

Greifswald-Insel Riems, Germany 

BHV-1 (IBR/IPV) control is based on marker vaccination and/or 

removal of infected animals which is mainly depending on the initial 

BHV-1 sero-prevalences. A field-infected animal has to be considered 

over its total lifespan as a latently infected carrier and as a potential 

source of infection. Therefore, BHV-1 diagnostics is crucial and often 

confronted with exceptional demands and requirements. For the 

sensitive and specific detection of BHV-1 antibodies, three different 

types of marker-independent tests are available: glycoprotein B (gB)blocking 

ELISAs, indirect ELISA tests, and the serum neutralisation 

test (NT). However, the gB-blocking assays represent the most 

IBR Workshop 

Angora, Beetal and Daira Din Panah) maintained at Rakh Khaire 

Wala Farm District Layyah. All the sera samples were first examined 

through Rose Bengal Plate Test (RBPT) and the samples found 

positive through RBPT were further confirmed through Serum 

Agglutination Test (SAT). 

Results: The overall prevalence of brucellosis was found to be 11% 

and 9.8% through Rose Bengal Plate Test and Serum Agglutination 

Test, respectively. Overall the females showed maximum prevalence of 

brucellosis in all breeds by RBPT (11.34%) and SAT (10.18%). Teddy 

goats showed the highest prevalence of brucellosis followed by Pak 

Angora, Beetal and Daira Din Panah. 

Discussion: This was a large scale screening of sera and in such cases 

the RBPT is an excellent tool and is the recommended method. This 

test has been widely employed in many successful Nationals 

Brucellosis Eradication Programmes. Based on the results, it can be 

concluded that the high prevalence of brucellosis is present in study 

area. Treatment of infected sheep and goats with antibiotics should be 

discouraged because the antibiotics may appear in the human food 

chain and this would be disastrous for the milk industry. Instead, efforts 

should be directed towards controlling and eradicating brucellosis from 

small ruminants by adopting integrated control strategies. Serological 

testing and slaughter of the animals that react positively with Brucella 

antigens successfully eradicated brucellosis in several countries. 

Vaccination against Br. melitensis may also be helpful in reducing the 

number of infected flocks and ultimately permit brucellosis control. 

Key words: goat, brucellosis, Pakistan 

sensitive test systems. In non-vaccinated animals, one ELISA system is 

commonly used as a screening test. In doubtful cases, a second ELISA 

principle can be employed as a confirmatory test. In contrast, BHV-1control 

according to the marker concept is based on vaccination with 

glycoprotein E (gE)-deleted marker vaccines. The marker-diagnostics 

is used to confirm the absence of gE-specific antibodies in "BHV-1 

(gE-antibody) free" animals and to detect gE-antibodies in cattle 

infected with wild type virus. Therefore, determination of absence or 

presence of antibodies directed against gE is the only tool to 

discriminate marker-vaccinated from wild type virus infected animals. 

In addition, gE-blocking ELISAs are at the moment the only available 

test system for the detection of gE-specific antibodies. The possibilities 

and limitations of the diagnostic systems for BHV-1 will be presented 

and discussed, e.g. problems of the different sensitivity levels of 

conventional and gE-marker tests, the failure of testing milk samples 

for gE-antibodies as well as several phenomena like the “vaccinationphenomenon” 

and the “fresh sample-phenomenon”. It will be 

demonstrated that the choice of the appropriate test system and the 

establishment of an internal batch control are prerequisites of the 

optimal performance of BHV-1-ELISA systems. In addition, the 

reasons for the occurrence of so-called “pseudo-vaccinees” - animals 

that are not marker vaccinated but react clearly positive for BHV-1 and 

negative for gE - will be analysed. Finally, the lack of confirmatory 

marker tests must be compensated so far by evaluation of the 

epidemiological plausibility of the test results and by emphasising 

diagnosis on the herd level. 

Key words: IBR, BHV-1, diagnostics, marker tests 

242 Prevalence and Risk Factors for Bovine Herpesvirus 1 

Infection in Cattle Herds from Parana State, Brazil 

J. Dias, A. Alfieri, J. Ferreira Neto, E. Muller, A. Alfieri 



The Brazilian cattle herd is constituted for approximately 205 million 

heads. The Paraná state, located in southern region of Brazil, is 

characterized by a developed livestock of beef and dairy cattle, with 

approximately 10.5 million heads. Throughout Brazil, the bovine 

herpesvirus (BoHV) infection is related to reproductive problems. 

Vulvovanigitis and meningoencephalitis are other clinical outcomes 

observed in infected herds. Serological surveys have showed a high 

Abortion – IBR Workshop 323

frequency of animals with BoHV antibodies around the country. This 

virus has also been isolated from young and adult animals with 

different clinical signs. The aim of this study was to evaluate the 

prevalence and risk factors involved with BoHV-1 infection in 

unvaccinated adult females of herds from Paraná state. The sampling 

was carried out in two stages. Firstly, the primary units were defined 

by an aleatory selection of a pre-established number of herds. In 

addition, from the primary units, in an aleatory way, it was sampled a 

pre-established number of animals (secondary units), representing the 

sanitary state of the herd (Herdacc ® version 3 software, University of 

Guelph). The calculation of the number of herds was determined by 

the degree of confidence of the result (5%), by the level of precision, 

and for the value of the expected prevalence (50%). Blood samples 

were collected from 14.083 females up to 24 months from 2018 

unvaccinated herds. Serum samples were tested for antibodies against 

BoHV-1 by using an indirect ELISA. The epidemiological 

questionnaire was applied to all the selected farms. One thousand 

four hundred and eighty one out of the 2.018 herds were positive for 

BoHV-1, representing a positive herds prevalence of 71.3% [69.3- 

73.3%]. Herds of beef cattle (OR= 1.58; IC: 1.12-2.23), natural 

service (OR=1.48; IC: 1.02-2.14), purchase animals (OR=1.90; IC: 

1.52-2.37), the rent of pasture (OR=2.24; IC: 1.51-3.33), existence of 

calving pens (OR=1.56; IC: 1.20-2.03) and history of abortion in the 

last 12 months (OR=1,45; IC: 1.08-1.95) were identified as risk 

factors for BoHV-1 infection in the multivariate logistic regression. 

These results indicate that BoHV-1 infection is widespread in the 

Paraná state and that factors related to herd characteristics and 

management are associated with the infection. 

Brazil Financial support: CNPq, CAPES, FINEP and FAP/PR 

Key words: BoHV-1, epidemiology 

243 Bovine Herpesviruses in Argentina: Present and Future 

S. Romera 1 , M. Vena 2 , A. Sadir 1 

1 Instituto Nacional de Tecnología Agropecuaria-INTA, Instituto de 

Virología, Buenos Aires, Argentina 

2 Biogenesis Bago S.A., Reserch & Development, Garin, Argentina 

Both BoHV-1 and BoHV-5 co-circulate in Argentina. Field strains 

of BoHV-1 and BoHV-5 were isolated from dairy and beef cattle 

herds. This fact may complicate future BoHV-1 control 

programs. BoHV-1 causes infectious bovine rhinotracheitis (IBR) 

and is reported around the world. BoHV-5 is a neurovirulent virus 

that causes fatal meningoencephalitis in young cattle, and is 

commonly reported in South American countries but only 

sporadically in other continents. The seroprevalence of BoHV-1 in 

Argentina ranges from 25% to 85%, serological results have been 

obtained by seroneutralization assays. Cross-reaction response 

between BoHV-1 and BoHV-5 was reported, then the true 

prevalence of BoHV-1 and BoHV-5 remains unclear. From 1981 up 

to now, fatal cases of neurological disease in calves associated with 

BoHV infection have been reported in the central region of 

Argentina. Up to now, BoHV-5 Argentinian field isolates were 

characterized by clinical presentation and restriction endonuclease 

analysis (REA) performed on virus isolation. Vaccination is a 

common strategy of control. In Argentina the vaccination against 

BoHV-1 is not mandatory and only inactivated vaccines are 

accepted. Although a control and eradication program has not yet 

been implemented, INTA’s Virology Institute has developed a 

marker vaccine to be used in future control programs. The 

development consisted of a gE deleted strain of BoHV-1, which was 

obtained by removing the complete ORF of the gene encoding 

glycoprotein E. Bovines were vaccinated with single and combined 

inactivated versions and live attenuated version along with bovines 

vaccinated with a commercial vaccine with wildtype BoHV-1. The 

marker vaccine induced serum and mucosal antibodies and allowed 

the differentiation between infected and vaccinated animals. In all 

the cases, vaccinated groups were protected against challenge with 

wt Los Angeles strain. On the other hand animals vaccinated with 

BoHV-1 gE- and challenged with BoHV-5 Argentinian strain 

(A663) were protected. Safety and Efficacy tests were satisfactory 

for both inactivated and attenuated live vaccines. In addition 

discriminating infected from vaccinated animals was possible by 

means of a commercial gE- ELISA test. It is concluded that INTA’s 

gE- marker vaccine is a safe and potent immunogen against BoHV-1 

and potencially against BoHV-5 and may be a useful tool for 

eradication programs in bovine herds. 


Key words: BoHV-1; BoHV-5, IBR, meningoencephalitis, marker 

vaccine 

244 IBR in France: from Voluntary Holding Certification to 

Eradication 

H. Petit 

FNGDS, PARIS, France 

In 1996, was created in France the “Association for Animal Health 

Certification” (ACERSA), in order to provide official certification to 

voluntary holdings, regarding diseases that are not under national 

regulations. The actual disease it was created for was IBR, mainly 

for export reasons. Members of ACERSA are breeders - represented 

by the GDS (animal health groups) -, veterinary surgeons 

associations and others such as laboratories, etc. The association is 

quality based and officially recognised by the French authorities.As 

for IBR, the main holding qualification is “IBR free”, based on 

regular testing of animals within the herd (either on bulk tank milk or 

blood serum), in association with adequate biosecurity measures 

regarding animals purchased, herds gatherings, etc. Since 1997, the 

number of “IBR free” holdings regularly progressed up to more than 

50 % in 2007.In addition to this certification system, the GDS 

national federation decided in 2004 to apply to the French authorities 

for a national regulation, in order to make IBR control compulsory. 

In November 2006, a law was adopted, specifying that all cattle 

holdings in France should be controlled at least once a year (using 

either milk or blood testing). All cattle purchased also have to be 

tested, and any positive animal has to be vaccinated within 2 months 

with proper life long boosters.This regulation enhanced to calculate 

national herd prevalence in 2007: 11 %. This proves that over 10 

years certification allowed a considerable improvement of the 

epidemiological situation of IBR in France: back to the 90’s, 

regional herd prevalence between 40 and 60 % (or even more) was 

not rare! But there are still important differences between regions, 

some like Brittany now being nearly free, whilst in other areas 

prevalence rates between 25 and 50 % still can be found. Generally 

speaking, regions with important dairy production show better 

situations than predominant beef production areas. On average, less 

than 2 % of the French cattle are vaccinated against IBR.Regarding 

E.U. regulations, France now plans to apply for inscription to annex 

I of decision 2004/558/CE, in order to obtain additional guarantees 

against IBR, and have its “free holding” status recognised by the 

European authorities. These measures, in addition to the fully 

operational French national system, should allow fulfilling the GDS 

medium term objective: BHV1 eradication. 

245 An Update on IBR Control in Italy 

S. Cavirani 

University of Parma, Animal Health, Parma, Italy 

BoHV-1 infection is widespread among cattle reared in Italy. 

Serological data on 1,650 dairy herds located in different Italian 

areas showed a seroprevalence to BoHV-1 of 46%. Diagnostic 

activity on 225 respiratory disease outbreaks occurring in dairy 

cattle in the period 2003-2007 demonstrated the involvement of 

BoHV-1 in 12% of the cases. On the basis of the clinical score, the 

most severe IBR outbreaks have been observed during the summer 

season. Despite a wide consciousness regarding the losses caused by 

BoHV-1 infection, only some areas of Italy are involved in official 

eradication programs. In particular, the province of Bolzano is 

officially free; compulsory eradication programs are in progress in 

the provinces of Trento, Belluno and in the Friuli region; voluntary 

eradication programs involve Lombardia, Veneto, Piemonte and 

Lazio regions. The main points of the eradication programs are: only 

dairy herds involved; certification of “free herd” status; use of 

marker vaccine only; herd monitoring (gE antibody detection on 

serum samples or bulk milk); gE positive animals sold out of the 

region. The use of vaccine is promoted by authorities independently 

form the herd status. At present the epidemiological role of beef 

cattle and buffaloes herds is of concern. Data regarding the 

vaccination procedure to IBR in Italy has been collected by a 

questionnaire carried out on 6,450 dairy herds during the period 

2002-2007. Overall vaccination involved 48% of the herds. In the 

areas without eradication program, traditional combined vaccines 

are widely applied. However, the trend shows an increase of marker

vaccine, mainly live vaccine. Due to the lack of combined vaccine 

containing IBR marker, the use of IBR marker vaccine 

simultaneously or concurrently with other vaccines is applied, 

mainly in large dairy herds.During the period 2000-2007, the follow 

up of the eradication has been performed in 232 dairy herds. The 

herds applied vaccination with marker vaccines without culling of 

seropositive animals and a mean replacement ratio of 30%. On the 

whole 88 herds achieved the eradication. The mean time to reach the 

eradication ranged from 3 years and 8 months to 6 years and 10 

months depending from the herd size and the starting 

seroprevalence. During the period of observation, in 32 (14%) of 232 

herds we have detected seroconversion to gE but in only 1 case the 

seroconversion was associated to clinical signs related to BoHV-1 

infection. 

246 Epidemiology and Eradication of the BHV1-Infection by the 

Example of Selected Farms in Lower Saxony 

J. Teuffert, K. Wiedl, F. Conraths 

Friedrich-Loeffler-Instutute, Department of Epizootiology, Berlin, 

Germany 

Objectives of study: A case-control study was conducted over two 

years (February 2004 - February 2006) in five selected administrative 

districts (Landkreise) in Lower Saxony to determine potential risk 

factors for the eradication of BHV1. 

Materials and Methods: Fourty-two control farms (cattle negative for 

BHV1) and 43 farms with an ongoing BHV1-eradication program 

(positive for BHV1; case farms) were chosen and studied on the basis 

of defined criteria. Information and data were collected regarding farm 

management, the BHV1 control and, if applicable, the BHV1 

immunisation strategy. Moreover, data on milk production and fertility 

were analysed for individual animals as well as herds. For this purpose, 

the BHV1 test protocols and vaccination lists that had been collected 

from various farms by the local veterinary authorities were compared. 

Finally, the farms were visited and the farmers interviewed using a 

standard questionnaire. 

Results: The statistical analysis revealed significant differences 

between the farm categories for 19 of the operational and 

management variables. In the evaluation of the data collected on 

individual infected animals, the distribution of age classes at the 

time of the first positive test result and the duration the infected 

animals remained on the farms until slaughter proved to be 

significantly different between case and control farms. In particular, 

the analysis of the vaccination strategy showed that the following 

factors differed between BHV1-negative and positive farms: the 

number of animals with proper basic immunisation, the distribution 

of the number of vaccinations administered in given intervals, the 

proportion of infected but unvaccinated animals, and the time lapse 

between the date of the first BHV1-positive test results and the 

measures taken. The differences in milk production and fertility 

between case and control farms, however, were not significantly 

different. Also, no significant difference was found in the 

performance of BHV1-positive and BHV1-negative cattle. 

Conclusions: In summary, the results of this study lead to 

recommendations on vaccination, care and feeding, transportation, 

trade and documentation in cattle farms to improve BHV1 control. 

Key words: BHV-1, epidemiology, risk factors, case control study 

247 Evaluation of the Probability of Transmission of a Live 

BoHV-1 Marker Vaccine to Sentinels after Intranasal or 

Intramuscular Application 

B. Makoschey 1 , J. Patel 2 , M. Beer 3 

1 

Intervet-Schering-Plough, Int. Marketing, Boxmeer, Netherlands 

2 

JAS Biologicals Ltd, Int. Marketing, Cambridge, United Kingdom 

3 

Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, Insel 

Riems, Germany 

Objectives: Live marker vaccines against bovine herpesvirus type 1 

(BoHV-1) infections are widely applied. The probability of 

transmission vaccine virus to sentinels after intranasal or intramuscular 

application should be determined. 

Materials and Methods: Two studies have been performed to 

investigate the transmission of a live BoHV-1marker vaccine (Bovilis ® 

IBR marker live, Intervet) virus after combined intramuscular (IM) and 

intranasal (IN) vaccination. A group of five (1 st study) or six (2 nd study) 

BoHV-1 negative calves was vaccinated with a 10x overdose of the 

vaccine IN and IM. The animals were housed together with 5 (1 st 

study) or 3 (2 nd study) unvaccinated calves during 2 weeks. Nasal swab 

samples were taken daily and tested for BoHV-1 vaccine virus. In a 3 rd 

study, it was established, whether animals vaccinated IM with the same 

vaccine become viremic and / or excrete vaccine virus with nasal 

discharge. Five animals were vaccinated once with an overdose via the 

IM route. Nasal swab samples were taken daily for 11 days. Blood 

samples were taken three times a week during the first two weeks and 

then once a week until four weeks after vaccination. The nasal swab 

samples and the blood samples were tested for BoHV-1 in a virus 

infectivity assay. In addition, a polymerase chain reaction (PCR) 

specific for BoHV-1 DNA was performed on the blood samples. 

BoHV-1 neutralising antibody titers were determined in the sera taken 

prior to the vaccination and four weeks after immunisation. 

Results: In the 1 st study, 1/5 in contact animals excreted a minimal 

amount of vaccine virus during 1 day and developed a BoHV-1 

antibody response at very low titer. All remaining sentinels in the 1 st 

and 2 nd study did not excrete vaccine virus and remained seronegative. 

In the 3 rd study, all animals were successfully vaccinated as judged by 

the development of BoHV -1 neutralising antibodies. However, all 

nasal swab samples were tested negative for vaccine virus, and all 

blood samples were found negative for BoHV-1 virus and BoHV -1 

specific DNA. 

Conclusions: It can be concluded that transmission of the vaccine virus 

to sentinels is very limited after intranasal vaccination and that no 

vaccine virus was excreted with nasal discharge after IM vaccination, 

nor could viremia of vaccine virus be detected. It is recommended to 

apply the tested BoHV-1 marker live vaccine by the IM route in 

situations where excretion of vaccine virus is undesirable. 

248 Evolution of the Net Reproductive Ratio of Bovine Herpes 

Virus Type 1 Infection in Hyperimmunized Dairy Herds: a 

Longitudinal Study in the Field 

M. Dispas 1 , R. Soares Magalhaes 2 , M. Pearce 3 , A. Dupont 1 , 

E. Thiry 4 , P. Kerkhofs 1 

1 

Veterinary & Agrochemical Research Centre, Virology, Brussels, 

Belgium 

2 

Royal Veterinary College, Epidemiology, London, United Kingdom 

3 


4 

Faculty of Veterinary Medecine-University of Liege, Virology, 


BoHV-1 control programmes have existed in Europe for 10 years. 

When seroprevalence is high, hyperimmunization of cattle is usually 

recommended to maximise virological protection. Marker vaccine 

usage has been promoted to allow discrimination of vaccinated cattle 

from cattle infected by a wild virus. Experimental studies show that 

effective immunity can be obtained by administration of live then 

inactivated vaccines (P1) but use of this protocol has in the field has 

not been reported. A 28-month longitudinal field study was 

conducted to compare the efficacy of protocol P1 with sole use of 

inactivated vaccine (P2), and the usual vaccination protocol defined 

as non intervention group (NIG). The study comprised 6 crosssectional 

serosurveys of all cattle on farms enrolled in P1 (n=10), P2 

(n=10) and NIG (n=16). New BoHV-1 infections were defined as gE 

seroconversion. Vaccinations and blood sampling followed the same 

schedule, providing 5 chronologically sequential time blocks: from 

winter 1, summer 1 and so on to winter 3. Mean net reproductive 

ratio (R) was calculated for each farm over the whole study and 

within each time block to quantify transmission of BoHV-1 in partly 

susceptible populations. Two methods were used: martingale 

estimation (R mart) and a linear mixed model using a log link and 

Poisson error distribution (R glm). Nine farms in P1, 8 in P2 and 13 in 

the NIG contributed to the calculation of R values. The mean R marts 

were, respectively, estimated to be 0.49, 0.61 and 0.70. Only the 

R marts for P1 and P2 did not include the value 1 in their confidence 

interval. In these groups BoHV-1 infection would fade out, but 

would persist in NIG. Mean R glms were smaller when infectiousness 

was presumed to be 3 days (respectively 0.23; 0.22 and 0.28 for P1, 

P2 and NIG) compared with 7 days (0.55; 0 .52 and 0.64) or 10 days 

(0.78; 0.74 and 0.92). In each group and respectively for the 5 time 

blocks, within each group, seasonal changes were observed: P1, 

0.64; 0.00; 0.37; 1.06 and 0.53; P2, 0.70; 0.77; 0.34; 0.73 and 0.41; 

NIG, 0.70; 0.61; 0.88; 0.49 and 0.93. Only P1 and P2 showed R mart 

values significantly smaller than 1, but not consistently. These 

IBR Workshop 325

esults, supported by a decrease in gE seroprevalence observed in the 

hyperimmunized herds (P1 and P2) and the decreasing number of 

herds affected by BoHV-1 infections show that both these protocols 

are helpful in BoHV-1 control and are more efficient than the usual 

vaccination protocols applied in the field. 

Key words: BoHV-1, hyperimmunization, gE marker vaccine, net 

reprodcutive ratio 

249 Evolution of the Net Reproductive Ratio of Bovine Herpes 

Virus Type 1 Infection in Hyperimmunized Dairy-beef Mixed 

Herds: a Longitudinal Study in the Field 

M. Dispas 1 , R. Soares Magalhaes 2 , M. Pearce 3 , A. Dupont 1 , 

P. Kerkhofs 1 , E. Thiry 4 

1 

Veterinary & Agrochemical Research Centre, Virology, Brussels, 

Belgium 

2 

Royal Veterinary College, Epidemiology, London, United Kingdom 

3 


4 

Faculty of Veterinary Medecine-University of Liege, Virology, 


BoHV-1 control programmes have existed in Europe for 10 years. 

When seroprevalence is high, hyperimmunization of cattle is 

usually recommended to maximise virological protection. Marker 

vaccine usage has been promoted to allow discrimination of 

vaccinated cattle from cattle infected by a wild virus. Experimental 

studies show that effective immunity can be obtained by 

administration of live then inactivated vaccines (P1) but use of this 

protocol has in the field has not been reported. A 28-month 

longitudinal field study was conducted to compare the efficacy of 

protocol P1 with sole use of inactivated vaccine (P2), and the usual 

vaccination protocol defined as non intervention group (NIG). The 

study comprised 6 cross-sectional serosurveys of all cattle on farms 

enrolled in P1 (n=10), P2 (n=10) and NIG (n=16). New BoHV-1 

infections were defined as gE seroconversion. Vaccinations and 

blood sampling followed the same schedule, providing 5 

chronologically sequential time blocks: from winter 1, summer 1 

and so on to winter 3. Mean net reproductive ratio (R) was 

calculated for each farm over the whole study and within each time 

block to quantify transmission of BoHV-1 in partly susceptible 

populations. Two methods were used: martingale estimation (R mart) 

and a linear mixed model using a log link and Poisson error 

distribution (R glm). Ten farms in P1, 11 in P2 and 15 in NIG 

contributed to the calculation of R values. Mean R marts were, 

respectively, 0.79, 0.44 and 0.68. There was no significant 

difference between groups. R mart values for P2 and NIG did not 

include the value 1 in their confidence interval.Mean R glms were 

smaller when infectiousness was presumed to be 3 days 

(respectively 0.14; 0.06 and 0.34 for P1, P2 and NIG) compared 

with 7 days (0.32; 0 .13 and 0.79) or 10 days (0.45; 0.19 and 1.13). 

In each group, respectively, for the 5 time blocks, seasonal changes 

were observed: P1 (1.02, 0.46, 0.84, 0.74 and 1.05); P2 (0.45, 0.66, 

0.38, 0.21 and 0.16); NIG(0.67, 0.63, 0.58, 0.87 and 0.64). Only P1 

and P2 had R mart values significantly smaller than 1, but not 

consistently. With the exception of S1, P2 always had the lowest 

R mart value.These results, supported by a decrease in gE 

seroprevalence observed in the hyperimmunized herds (P1 and P2) 

and the reduced number of herds affected by BoHV-1 infections 

show that both these protocols are helpful in BoHV-1 control and 

are more efficient than the usual vaccination protocols applied in 

the field. 

Key words: BoHV-1, gE marker vaccine, net reproductive ratio, beef, 

dairy 

250 Modelling the Impact of Vaccination on Bovine Herpesvirus 1 

Eradication on Dairy Farms 

L. Matthews 1 , M. Pearce 2 , I. Vangeel 2 

1 

University of Glasgow Veterinary Faculty, Glasgow, United 

Kingdom 

2 

Pfizer Animal Health, Sandwich, United Kingdom 

Many European countries have embarked on the eradication of 

bovine herpesvirus 1 (BoHV-1) from their national cattle herds and 

are using BoHV-1 vaccines in their programmes. Our aim was to 

investigate the impact of BoHV-1 vaccination on the prevalence and 

persistence of BoHV-1 on dairy farms, including hypervaccination 


i.e. primovaccination of young animals followed by 6-monthly 

booster vaccinations. We developed a stochastic, individual animalbased 

model of the herd and management structure and animal 

turnover of a typical dairy farm. Animals were categorised into four 

core management groups: milking cows; dry or calving cows; 

replacement heifers; and calves. The model was used to simulate 

changes in the prevalence of BoHV-1 infection and the persistence 

of BoHV-1 infection on a farm under three vaccination schedules: no 

vaccination; hypervaccination; and hypervaccination of all cattle. 

The impact of these vaccination schedules was examined under 

different scenarios defined by the following parameters: initial 

BoHV-1 prevalence, viral reactivation rate in latently infected cattle; 

the typical lifespan and length of time spent by an animal on the 

farm; herd structure and management; management group size; 

mixing of animals including movement of animals between the 

management groups and the importation of replacement animals, and 

the probability that imported cattle are infected. Estimates of the 

basic reproduction ratio obtained from data collected in field studies 

were also used. Several key findings have been demonstrated. 

Outcomes can be quite variable, even for the same initial 

parameterisation, and the model highlighted the sensitivity of 

outcome to key parameters for which there is still uncertainty such as 

transmission rate, duration of immunity after infection and 

reinfection, the risk of vertical transmission, rate of reactivation and 

the relative infectiousness of animals after reactivation. Reactivation 

can result in protracted outbreaks, which is exacerbated as herds 

become larger, and the number of animals in a group is an important 

determinant of time to extinction. Vaccination can have a substantial 

impact on the expected time to extinction of BoHV-1 in a herd. Even 

with highly efficacious vaccines, minimal extinction times of 4 to 5 

years can be expected because of time taken to remove latently 

infected animals under usual management practices. The model 

demonstrates that hypervaccination is more effective than just 

vaccinating adult cattle. 

Key words: IBR, BoHV-1, modelling, vaccination, eradication 


1054 Seroprevalence of Bovine herpesvirus 1 (BHV-1) infection 

among Estonian dairy cattle 

K. Raaperi 1 , A. Viltrop 1 , T. Orro 1 , K. Must 2 , K. Mähar 2 , I. Nurmoja 2 

1 Estonian University of Life Sciences, Institute of Veterinary 

Medicine and Animal Science, Tartu, Estonia 

2 Estonian Veterinary and Food Laboratory, Tartu, Estonia 

Objectives: BHV-1 infection is endemic in Estonian cattle. In national 

IBR survey based on herd bulk milk samples in 2004, 16.4% of herds 

were BHV-1 antibody (ab) positive. The objectives of our study were 

to estimate the herd prevalence and within herd prevalence of the 

infection in Estonian dairy cattle. The age profile of the infection in 

heifers was investigated and the sensitivity of the bulk tank milk 

(BTM) ELISA test to detect infected herds was estimated 

Material and methods: 77 dairy herds were randomly selected 

considering their BHV-1 status (positive or negative) and size. 

Representative sample of cows and heifers from 6 months old were 

tested from blood with a commercial ELISA. A BTM sample from 

every milk tank of the herd was tested. 1205 BTM samples from the 

laboratory of the Animal Recording Centre (ARC) were also tested for 

BHV-1 ab-s. 

Results: 19% of BTM samples collected from ARC were positive to 

BHV-1. The sensitivity of the ELISA test to detect infected herds using 

BTM samples was estimated as 70% and the specificity 100% and the 

true prevalence was calculated as 27% (95% CI 25.8-28.4). Assuming 

the same sensitivity and specificity of the test in 2004 survey, the 

increase in national herd prevalence was statistically significant. 

Within herd prevalence in infected herds of the study population was 

on average 43.5% (range 1-100%; median 30%). The average within 

herd prevalence increased with herd size. Prevalence in herds with 20- 

99 cows was 2-63% (av. 19.7%, med. 5%). In herds with more than 400 

cows, prevalence range was 1-91% (av. 54.4%, med. 46.5%). In 80.5% 

of infected herds the prevalence was 40% and in 19.5% of infected 

herds the prevalence in heifers was 60%. The prevalence in heifers 

increased with age (OR between one month of age was 1.12 (95% CI 

1.05-1.2; p

Conclusions: Herd prevalence of BHV-1 infection has increased 

compared to 2004. Within herd prevalence increased with herd size. 

Based on the prevalence in heifers the infected herds can be divided 

into two groups: low and high prevalence herds. In most herds the 

spread of the infection is slow and young stock becomes infected 

291 On-farm Teaching of Veterinary Students - Experiences and 

Perspectives 

I. Klaas, R. Jorgensen, C. Enevoldsen 

University of Copenhagen, Faculty of Life Sciences, Department of 

Large Animal Sciences, Frederiksberg, Denmark 

Rapidly increasing herd sizes in Denmark increase the demand for 

veterinarians, who are competent to apply a systematic approach and 

have solid knowledge on cow-level, herd-level and business matters. 

At the same time, most of the veterinary students grow up in cities 

with little or no contact to cattle and dairy production systems and 

probably little interest in becoming a cattle veterinarian. Both the 

industry’s demand and the students’ background emphasize the 

increased need for adapting the clinical teaching to meet the 

demands of modern dairy production. With on-farm teaching using 

the farm as a case the students can learn and apply their knowledge 

under real life conditions (contextual learning). We established a two 

weeks training course for veterinary students 1-2 years before 

graduation and after basic training in hospitals. There are three farm 

visits in typical commercial dairy farms with herd sizes ranging from 

50 to 260 cows. The aim of the course is that the students achieve 

competencies necessary for their authorization as veterinarians 

(diagnosing and treating diseases of cattle). Further, they have to 

achieve the basic competencies required to work according to 

officially approved herd health advisory contracts. From September 

2006 to May 2007 six groups of students passed the courses. Typical 

elements and assignments of the farm visits are: description of the 

system (housing, feeding, and management of different age groups), 

clinical examinations of acute patients, systematic clinical 

examination of fresh cows, cows to be dried off, calves, a random 

sample of milking cows and description of milking procedures. In 

interviews with the owner or farm manager the students train to 

communicate and gain knowledge on different types of management. 

All theoretical exercises are based on farm visits and aim to give an 

assessment of the health and welfare status of the herd. The 

assessment consists of the students’ results from the systemic 

clinical examinations, interpretation of herd data and management. 

Course evaluation and practical experiences show that farm-based 

teaching is an important and effective tool to improve and gain 

knowledge in a short period of time. Furthermore, on-farm teaching 

increases the interest of students in production animals, which could 

be important to ensure that enough cattle veterinarians can be 

recruited for cattle practice in the future. 

292 Recent Developments in Bovine Veterinary Education in 

Berlin with Students' Eyes 

K. Heenemann, C. Weber, K. Muller 

Clinic for Ruminants and Pigs, Faculty of Veterinary Medicine, Freie 

Universitaet, Berlin, Germany 

The veterinary education in Germany should be completed in five 

years and six months. It is divided into a pre-clinical and a clinical 

part. Contents and structure of the veterinary programme at the Freie 

Universität Berlin are regulated by the German Tierärztliche 

Approbationsverordnung (TAppV). The education in buiatrics starts 

in the 3 rd year with courses on clinical examination techniques. 

Lectures and clinical case demonstrations are held in the course of 

organ-centred modules. At the Clinic for Ruminants and Pigs most 

of the lectures include Blended Learning, increasing the active 

participation of the students. As the students are able to prepare for 

the lectures and clinical case demonstrations using the E-Learning 

Education Workshop 

through extended time period. The ELISA test used in the study was 

not efficient to discover herds with low and moderate prevalence 

(

international drivers of change. Those involved in veterinary 

education need to respond to these changes and be capable of 

anticipating and meeting the critical demands and needs of the 

agricultural sector and the wider community for a safe food supply in 

the 21 st century. Future success will be based on developing a 

critical mass of veterinarians with special skills, knowledge and 

training to effectively participate in herd health programmes; 

veterinarians who will be involved in the provision and certification 

of a safe food supply in the context of sustainable agriculture. 

294 Veterinary Education in the Netherlands: Ready for the 

Future 

B. Hietberg 

Utrecht University, Farm Animal Health, Utrecht, Netherlands 

In the Netherlands, there is one Faculty of Veterinary Medicine 

(FVM). The FVM is accredited by AVMA/CVMA. Each year 225 

students enrol either by weighted lottery or by a selection procedure. 

For the Farm Animal Health (FAH) track especially 50 to 70 

students are selected each year. The first 4 years aimed a more 

theoretical knowledge and insight into healthy and diseased animals. 

In this phase 25% of these 4 years is spent on FAH. This is educated 

by spending extra time on FAH specific literature, exercises and 

exams. The last 2 years consist of clinical rotations and research. The 

first 30 weeks are uniform clinical rotations and exist of clinical and 

extramural training at all departments. Furthermore, every student 

performs a scientific research project for at least 3 months. The last 

year of the study is solely spent on FAH education. This consists of 

teaching in the clinic, the ambulatory practice of the faculty, 

extension practice and externship. Much attention in this track is 

paid to the food-producing-chains, herd health and farm production 

management, Veterinary Public Health and animal welfare. In my 

personal experience, there is a good interaction between studying the 

theory and getting practical experience. The strengths of the current 

curriculum are; the scientific approach, the physical facilities, the 

good possibilities for personal development in the veterinary world 

and the involvement of students in the evaluation and development 

of education. There is a wide variety of additional activities 

organized by many student societies. The student develops a good 

professional conduct, and a strong social and communicative skill. 

There is a good interaction between the students, the teacher and 

other faculty members. Due to the continuous growing quantity of 

new and easy available knowledge I believe the ready knowledge of 

the student is sometimes disadvantaged. In our curriculum, teachers 

are expected to be multifunctional, they have to teach inclusive 

assessment of professional, social and communicative skills of the 

student, perform research, and do clinical work. There are relatively 

few teachers who are master of all these disciplines. In conclusion 

the FVM in Utrecht is a great place to learn FAH with lots of 

possibilities. The FAH track is broad and aimed at the future. The 

FAH student is educated to be able to manage with emergency work, 

be an advisor to the farm owners and to control the Veterinary Public 

Health. 

295 Using Rich Media for Teaching Veterinary Obstetrics 

J. Scherzer, M. Buchanan, J. Moore, S. White 

University of Georgia, College of Veterinary Medicine, Athens, 

United States 

Objectives: Teaching of veterinary obstetrics is restricted to 

descriptions of birth and dystocia in text or, at best, in two dimensional 

graphical presentations. The creation of three-dimensional, interactive, 

digital animations will help students not only understand but also 

visualize processes during normal birth and dystocia. 

Materials and Methods: We have created rich-content, three 

dimensional animations that were designed to help students 

understand the process of normal birth and dystocia. This technology 

has been used successfully in our College of Veterinary Medicine to 

depict gastrointestinal diseases of the horse, and complex 

intracellular processes. The latter of which was cited as having a 

significant positive effect on students' understanding of intracellular 

pathways (1). The veterinary obstetrics project incorporates both 

linear animations and QuickTime VR objects. The animations are 

used to demonstrate the condition of normal birth as well as dystocia 

and the appropriate treatment. The QuickTime VR objects will be 


used to demonstrate the conditions as a static object that the students 

can manipulate and examine from all angles. These materials have 

been used in instruction of 4th year veterinary students. Students 

reported whether these materials enhanced their understanding of 

parturition and dystocia. 

Results: All students exposed to the three dimensional teaching 

materials reported a greater understanding of parturition and dystocia. 

Conclusions: Three dimensional animations of normal parturition and 

dystocia enhanced students' understanding of these processes, enabling 

them to more confidently assist in parturition and treat clinical cases of 

dystocia 

Key words: veterinary, obstetrics, three dimensional, animations 

296 The New Curriculum - Veterinary Medicine with Tracking in 

Food Animal Medicine - at the Vetsuisse-Faculty of the 

University of Berne, Switzerland 

L. Balet, K. Reinmann 

University of Berne, Food Animal Medicine, Berne, Switzerland 

Introduction: In Switzerland, the “new” curriculum has started in 

2004 and is offered at both Vetsuisse-faculties of the Universities of 

Bern and Zurich. This presentation covers Bern mainly, and focuses on 

the 4 th and 5 th year. 

Materials and Methods: Information has been collected from the 

office of student affairs and the EAEVE self evaluation report 2007. 

Results: Veterinary studies take a minimum of five years, but before 

starting, candidates have to pass an aptitude test for medical studies. 

During the 1 st year, the students learn the basic knowledge in biology 

and medicine. During the 2 nd and 3 rd years, the topics are divided in 

organ-focused and non-organ-focused units. In each organ-focused 

unit, students learn anatomy, physiology, special pathology, and 

clinical disciplines of the various species (pets, horses, food animals) 

of the respective organ system. For the two final years, the students 

choose among one of the following tracks: small animals, food 

animals, horses, VPH, research, or para-clinical disciplines. In 2007, 

50% (22) of the Bern students have chosen the “food animal” track. 

The 4 th year is structured as follows: 11 weeks of indepth VPH and 

general clinical topics for all students, 9 weeks of indepth training in 

the respective track, and 8 weeks of symptom-oriented casemanagement 

of all 3 clinical tracks for all students. Modes of 

learning may include a combination of literature studies, teamwork 

problem solving, hands-on practical training, computer based 

learning and attending few review lectures. During the 5 th year, 

students are on clinical rotation, including two 4-week-practical 

externships and 11 weeks of indepth training in the institute or clinic 

of their thematic priority. Students in the food animal track, spend 

their clinical training in the swine clinic, the ruminant clinic and the 

division of herd health management. The ratio of food animals 

students to food animal cases in Bern is currently 1:122. The clinical 

rotation often allows a 1 student to 1 instructor ratio in clinical 

training. 

Discussion: The result of this new curriculum is that students are 

specialized to some degree already at the end of their studies. They 

acquire practical skills and indepth knowledge in their track of choice, 

but this is at the expense of other clinical fields. This may impede 

young veterinary graduates to find a job in a field other than that of their 

specialisation. 

297 Future changes in the Supply of Veterinary Services could 

affect the Welfare of Livestock in Urban Areas of the United 

Kingdom 

S. Borsberry 

Veterinary Practitioner, Private Veterinary Practice, Warwick, 


Approximately 2,900 dairy cows, 1,700 beef cows, their followers, 

25,000 breeding ewes and their lambs are serviced by a large mixed 

practice in the West Midlands.There are 5 large animal veterinary 

surgeons are employed solely to treat equines and livestock. A two 

year period (1 st January 2006 to 31 st December 2007) of large animal 

visits was analysed. There was a total of 12,054 visits of which 9245 

and 2809 related to equine and farm animals respectively.Out of 

hours visits are included in the visits of which 735 related to equines 

and 298 to production animals. Further analysis shows that the 

majority of the farm visits and procedures were performed by one

clinician (B).Of the farm visits B carried out 92% (2576 of 2809), 

90% (121of 135) bovine obstetrical procedures and all (n3) of 

operations for right sided displacement and torsion of 

the abomasum. All health plans and routine fertility visits to both 

dairy and beef cows were carried out by B. B has worked in this 

practice for 37 years, is due to retire and the farm clients are 

concerned as to the future supply of veterinary services. It is unlikely 

that the existing clinicians will gain the qualifications of B or that on 

retirement the practice will attract a clinician with similar experience 

and qualifications. The practice has a relatively small farm animal 

client base compared to many farm orientated UK practices, for 

example, an eight person practice in Cheshire looks after 30,000 

dairy cows and an eight person practice in Yorkshire looks after 

9000 dairy cows and 9000 beef cows.Equine emergencies tended to 

be limited to wounds, limb fractures and colic. The majority of 

wounds were dealt with by one of the clinicians; surgical colics were 

transported to a neighbouring horse hospital for further treatment 

and most limb fractures were euthanased. Bovine emergencies are 

dealt with on farm. This urban practice is not dissimilar to other UK 

practices with relatively small livestock bases. It is likely that such 

practices will cease to service the livestock industry and will be 

attended by specialist farm veterinary surgeons from a greater 

distance which may well compromise the welfare of individual 

animals requiring prompt attention. 

Key words: welfare, bovine obstetrics, veterinary services 

298 Teaching Beef Cow-Calf Production Medicine to Veterinary 

Students at Purdue University, USA 

W. Hilton 

Purdue University, School of Veterinary Medicine, Veterinary 

Clinical Sciences, West Lafayette, United States 

Purdue University began teaching Beef Production Medicine in 1999 

with an elective, lecture course in the second semester of the third of 

four years of veterinary school. The course is divided into the 

subjects of: Health, Records, Fertility, Marketing, Genetics, 

Nutrition and Environment. Actual herd histories are presented to 

the students who work in groups in a problem-based learning setting 

during class to develop a list of questions they would ask the client. 

The instructor provides answers to these questions and the students 

then formulate possible solutions to the owner’s concerns. During 

the fourth year of school, the students get an opportunity to do farm 

visits and actually see these farms that they have discussed in class. 

This way they can actually experience the entire business and see if 

their suggestions are workable options for the owner. The student 

feedback on the lecture course has been very positive and they rate 

the actual on the farm experience as critical to their long term 

retention of this subject. 

Key words: beef production medicine, teaching, problem-based 

learning 

299 Training in Reproduction within the Framework of the 

"Intensive Study Munich". Results of an Evaluation by 

Students of Veterinary Medicine 

E. Zeiler 1 , R. Martin 1 , J. Friker 2 , R. Mansfeld 1 , H. Zerbe 1 

1 University of Munich, Clinic for Ruminants, Munich, Germany 

2 University of Munich, Institute for Anatomy, Munich, Germany 

The "Intensive Study Munich" has been started at the Veterinary 

Faculty of the UM as a clinical rotations programme during the 

summer semester in 2005. It is provided for students of the 8th and 

9th semester to improve the clinical training programme within the 

veterinary curriculum. Passing the curriculum of the reproduction 

unit in the Clinic for Ruminants by the students takes 3.5 weeks 

concerning a clinical or an ambulatory/herd health management 

division. Participants are integrated in all daily routines of the 

clinical work getting lots of opportunities to improve their practical 

skills. In the ambulatory/herd health management division students 

visit dairy farms accompanied by a veterinarian. Additional to the 

clinical work there are practical-oriented lessons in andrology, 

obstretics, gynecology and herd health management. The efficiency 

and acceptance of training units concerning reproduction medicine 

and production medicine/herd health were evaluated by 211 

students, who had passed the training units of the Clinic for 

Ruminants. 30 random chosen evaluation sheets were analysed each 

from the clinical division and the ambulatory/herd health 

management division of the clinic. The evaluation sheets, providing 

grades from “1” (very good) to “5” (inadequate), included aspects of 

organisation of the training units, well being of the students as team 

members, effectiveness of lessons, support by teaching stuff and 

learning success. All aspects were mostly evaluated as “very good” 

or “good”. Overall learning success was evaluated as “very good” by 

61.3% and 60.0% of the participants of the clinical and 

ambulatory/herd health management division, resp. The better result 

of evaluation of the well being of the students as team members and 

the support by teaching stuff by the students in the ambulatory/herd 

health management division caused in an intensive collaboration 

between students and teaching veterinarians. By driving the whole 

day from farm to farm they grow to a team. On the other hand the 

visits of farms are often limited in time. In addition the farmers are 

present during the therapies causing restriction of the practical 

engagement of the students. So the training of practical skills in the 

clinical division was evaluated as more intensive than in the 

ambulatory/herd health management division. The results of this 

evaluation show that the new form of clinical education is very well 

accepted by the students. 


1051 Changes in Daily Gain of Weight with the Inclusion of 

Leaves of Agave Blue Tequilana Weber in the Food of the 

Beef Bullocks 

I. Ruiz, Y. Martínez, J. Orozco 

Universidad de Guadalajara, Biological Sciences, Tepatitlan Jalisco 

Mexico, Mexico 

The harvest of the agave head, which is used to obtain the tequila 

drink, produces the leaves (also called “pencas” which are left on the 

field to rot without any benefit for the agave producer who has also 

ruminants in the farm. The present trial was undertaken to assess the 

apparent digestibility of total mixed ration containing three levels of 

agave leaves, using bullocks (n = 9) bearing a harness to collect the 

feces. The experiment lasted 21 days of which the last 5 were to 

assess intake and feces production and sampling. The data obtained 

were analyzed (SAS 2005) as a randomly designed trial for three 

agave leaves levels (0, 20, 40%). As the agave leaves were increased 

in the feed the nutrients intake was augmented (P = 0.058), however 

the daily gain of weight was unchanged (P > 0.05). On the other 

hand, the digestibility of most of the nutrients was increased as the 

leaves of the agave plant were augmented (P < 0.05). Based on the 

findings, it could be concluded that the agave leaves can be used to 

up 40% in the beef bullocks. 

1052 Gynaecology, Obstetrics and Artificial Insemination 

Phantom 

SM. Zolhavarieh, HR. Zahiri 

Junior School of Veterinary Medicine, Bu-Ali Sina University, 

Clinical Sciences, Hamedan, Iran 

Introduction: The phantom of cow is a simulator developed to train 

veterinary students to palpate the bovine reproductive tract, to 

perform fertility examinations, to diagnose pregnancy and to perfom 

artificial insemination (AI) in the cow. The phantom uses natural 

organ, which allows student to touch the real condition. When being 

trained with this phantom, the student palpates real parts of the 

bovine reproductive tract. The teacher provides instruction and 

feedback while following the student's actions inside the phantom 

through of specific window. The procedures are difficult to teach 

and to learn using traditional methods and students need plenty of 

opportunities to practice to develop the required skills. When 

palpating cows hand movements are not visible, which means the 

student cannot copy what the teacher did and it is also difficult to 

provide effective guidance. The phantom has been developed as a 

supplement to existing training methods, which the natural organ is 

positioned inside a fibreglass model of the rear-half of a cow. 

Validating the phantom: Research work has been undertaken by 

writers to validate the phantom. It was important to establish that 

skills developed during phantom training transferred to the real task. 

Education Workshop 329

The performance of two groups of novice veterinary students was 

compared. One group of students were trained with the phantom, the 

other group received only traditional training. The results showed 

that the performance of the phantom trained group was significantly 

better than the other group. Therefore, the phantom provides an 

effective supplement to traditional training for the fundamental skills 

students need to learn: locating the uterus in the cow, full fertility 

examinations, diagnosing pregnancy or even putting a scanner in the 

right place. 

Key words: phantom, teaching, gynaecology, reproduction 

1053 Distance Education Programs for Specialization in Bovine 

Production 

E. Tellez, MA. Quiroz 

Universidad Nacional Autónoma de México, Facultad de Medicina 

Veterinaria y Zootecnia, Cattle Health Service, México D.F., Mexico 

The modern distance education programs started in the United 

Kingdom in the decade of the sixties, in the XX th century. In the 

following decade, similar studies were initiated in Spain and also in 

Mexico. Dr. Pablo González Casanova was dean of University when 

the Distance Education Program was instituted in the Universidad 

Nacional Autónoma de México, in the year of 1972. This distance 

education program offered by the university provided courses for 

college degrees. However, the Facultad de Medicina Veterinaria y 

330 Introduction to Veterinary Genetics 

C. Droegemueller 

University of Berne, Institute of Farm Animal Genetics, Bern, 


The huge spectrum of genetic phenotypes in cattle covers anomalies, 

disorders, diseases and also morphological traits. A detailed clinical 

and pathological description and examination of the phenotype is the 

most important prerequisite before initiating genetic studies. After 

the recognition of a new genetic phenotype it is recommended to get 

an idea about the possible mode of inheritance, e.g. simple 

monogenic or more complex polygenic, by pedigree based 

segregation analysis. Once there is evidence for a genetic component 

an appropriate number of DNA samples of cases and control is 

required to perform either targeted candidate gene or genome wide 

analyses. 

331 Defects of the Skeletal System: Congenital Spinal 

Malformations 

J. Agerholm 


Veterinary Pathobiology, Frederiksberg, Denmark 

Skeletal malformations are among the most commonly recognised 

congenital malformations in cattle and many of these have been 

reported in the spine. Spinal malformations may occur as part of a 

generalized bone disease or may only affect the spine. Some spinal 

defects are a part of a syndrome or the spinal lesion may induce 

secondary lesions in other tissue. Arthrogryposis is a common 

lesion associated with malformation of the spine. Chondrodysplasia 

and osteogenesis imperfecta are two generalized bone defects 

affecting the vertebrae. Several types of chondrodysplasia exist and 

these have traditionally been designated on the basis of their 

morphology or breed affiliation. However, recent progress in 

molecular biology has started to solve the riddle of 

chondrodysplasia thus forming the basis for a more appropriate 

terminology. Osteogenesis imperfecta is an inherited collagen 

defect associated with decreased strength of bones and other tissues 

having type I collagen as an important structural part. Although 

spinal lesions are a part of this disorder, lesions in other parts of the 


Genetics Diseases Workshop 

Zootecnia offered only specialization courses. This study system in 

Mexico was created due to the students´ demand for places in the 

university. However, the Facultad de Medicina Veterinaria decided 

to only implement the specialization courses which were integrated 

by veterinarians, pedagogues, educators, psychologists, designers 

and professionals in communication, mainly specialists on 

television and cinematography. These groups started the 

preparation of all the materials required for the courses, the 

programmed texts and the didactic guides required for the subjects. 

These latter materials were supported with slides and super 8 and 16 

mm films which would eventually be replaced with computer 

technology material. The teaching-learning binome is integrated by 

the students and veterinarians and their teachers which are called 

tutors. The teaching-learning activity takes place outside the 

university campus since the students are practicing veterinarians in 

their communities and have other responsibilities to their jobs and 

their families that make it impossible for them to go to 

specialization courses in the universities located in the big cities. 

Therefore, the tutors travel every other week to meet with the 

students in different states of the country. Study groups are formed 

and the students meet with the tutor. At the moment, there are six 

study groups in towns located in strategic points where the students 

have easy access. In the last fifteen years, more than 600 

professionals have concluded their specialization courses and 

received their diploma which is a much greater number of graduates 

by the traditional university courses. 

skeleton are generally more obvious. Several disorders associated 

with widespread vertebral malformation have been reported. The 

Complex Vertebral Malformation (CVM) syndrome has occurred in 

Holsteins worldwide. CVM is due to a single base mutation in the 

SLC35A3 gene, which is essential for normal segmentation of the 

spine. Recently, the brachyspina syndrome has been identified in 

the Holstein breed. The spine is abnormal short due to widespread 

malsegmentation and the growth is severely retarded (body weight 

around 10 kg). Affected calves also have internal organ defects as 

i.e. renal dysplasia and they are usually stillborn. Several local 

spinal defects have been reported but their inheritance has in most 

cases remained unsolved. Deviations from the normal course of the 

spine (i.e. scoliosis, kyphosis, lordosis, and torticollis) are rather 

common. Fusion of vertebrae might affect all levels of the spine, but 

atlanto-occipital fusion is the most frequently reported type. 

Absence of vertebrae is mostly reported as absence of lumbosacral 

vertebrae (Perosomus elumbus) or absence of caudal vertebrae 

causing shortening or absence of the tail. A specific syndrome 

exists in which absence of caudal vertebrae is associated with 

genital, urinary and alimentary defects. Absence of vertebrae might 

not be complete. Abnormal development of the vertebral arch is 

seen in spina bifida. 

332 Defects of the Skeletal System: Update on Syndactyly in 

Cattle 

A. Eggen 1 , S. Floriot 1 , A. Duchesne 1 , A. Ducos 2 

1 

INRA, Laboratoire de Génétique Biochimique et de Cytogénétique, 

Jouy-en-Josas, France 

2 

INRA, Station d'Amelioration Génétique des Animaux, Centre de 

Toulouse, Castanet-Tolosan, France 

Syndactyly in Holstein cattle is an autosomal recessive abnormality 

characterized by the fusion of the functional digits. This disorder has 

been previously mapped to the telomeric part of bovine chromosome 

15. We recently fine-mapped the syndactyly locus in Holstein cattle 

to a 3.5-Mb critical interval using a comparative mapping approach 

and an extended pedigree generated by embryo transfer. We reported 

genetic evidence for the exclusion of two genes previously suggested 

as candidates (EXT2 and ALX4) and described the identification of 

a doublet mutation in complete linkage disequilibrium with 

syndactyly in one gene of the critical interval: LRP4. Finally, based

on recent discoveries concerning the mouse mutants dan and mdig 

and a mouse knockout for Lrp4, we present solid evidence that the 

subsequent substitution in LRP4 exon 33 is a strong candidate causal 

mutation for syndactyly in Holstein cattle. 

333 Inherited Defects of the Brown Breed: Genetic Diseases of the 

Central Nervous System 

K. Doll 

University of Giessen, Clinic for Ruminants and Pigs, Giessen, 

Germany 

Bovine progressive degenerative myeloencephalopathy (PDME) 

or weaver syndrome, an autosomal recessively inherited 

neurological disease, was first observed in American Brown Swiss 

in 1973. Clinical signs usually begin at 6 months to 2 years. The 

progressive bilateral hindlimb weakness and proprioceptive deficits 

cause difficulty in rising and a weaving, hypermetric gait, and finally 

recumbency.Pathological findings include axonal degeneration in 

the white matter of the thoracal spinal cord, and myelin degradation. 

Additionally nerve fibers of the peripheral nervous system exhibit 

comparable mitochondrial alterations and identical alterations are 

encountered in skeletal muscles. Weaver syndrome can be 

considered as a primary axonopathy, and a mitochondria-related 

encephalomyolo-/myopathy, respectively. Bovine spinal muscular 

atrophy (SMA), an inheritable autosomal recessive neurological 

disease, is characterized by degeneration and loss of motor neurons 

of the spinal cord and motor nuclei of the brain stem, and 

denervation and neurogenic atrophy of the striated muscle. Electron 

microscopy reveals accumulation of neurofilaments and 

mitochondria in affected neurons. SMA was first described in 1988 

in calves of the Red Danish breed x American Brown Swiss. The 

progressive weakness with respiratory failure finally causes death in 

the affected animals. The disease shares many features with 

Werdnig-Hoffmann disease in man and the spinal muscular 

atrophies of other animals. Bovine spinal dysmyelination (SDM), 

an autosomal recessive neurological disorder, was first reported in 

new-born brown Swiss x Braunvieh calves in 1993 and in calves of 

the Red Danish Dairy breed x American Brown Swiss (1994). It is 

characterized by congenital lateral recumbency, opisthotonus, 

extension of the limbs, normal to increased reflexes, and normal 

alertness. At histological examination dysmyelination occurres in 

spinal tracts, affecting the ascending gracile funiculus, the ascending 

dorsolateral spinocerebellar tract, and the mainly descending 

sulcomarginal tract. Deficient myelin production, loss of myelin, 

consecutive axonal degenerations, and astrogliosis within these 

tracts are the histological characteristics. This primary myelination 

disorder differs from other hereditary neurological diseases in 

Brown Swiss and Braunvieh cattle, i.e. the weaver-syndrome and the 

spinal muscular atrophy. 

334 Inherited Defects of the Brown Breed: Arachnomelia 

A. Gentile 1 , S. Testoni 2 

1 


Italy 

2 

University of Padova, Department of Veterinary Clinical Sciences, 

Padova, Italy 

Arachnomelia (“spider-legs”) is a congenital abnormality of the 

skeletal system giving the animal a spidery look, and reported both 

in Brown and Simmenthal calves.The most important pathologic 

findings are: facial deformities (i.e. brachygnatia inferior and 

concave rounding of the dorsal profile of the maxilla), bone 

dolichostenomelia, angular deformities in the distal part of the hind 

legs, muscular atrophy and cardiac malformations. The bones of the 

legs appear to be more fragile than normal and spontaneous fracture 

during calving may injure the dam. 

335 Inherited Defects of the Brown Breed: Genetic Aspects 


University of Berne, Institute of Animal Breeding, Bern, Switzerland 

Four independent different genetic diseases were observed in Brown 

Swiss cattle. For all phenotypes an autosomal monogenic recessive 

inheritance has been described. In 1993 the first genetic disease in 

cattle, progressive degenerative myeloencephalopathy (Weaver 

disease), which is also associated with increased milk production, 

was genetically mapped to bovine chromosome 13. In 2001 the 

spinal dysmyelination (SDM) was mapped in crossbred American 

Brown Swiss cattle to bovine chromosome 11. In 2007 the first 

causative mutation for a genetic defect in German Brown cattle was 

reported for the neurodegenerative disease spinal muscular atrophy 

(SMA). The bovine FVT1 gene, encoding 3-ketodihydrosphingosine 

reductase, which catalyzes a crucial step in the glycosphingolipid 

metabolism, showed a perfectly SMA associated missense mutation. 

In 2008 a microsatellite marker based whole genome scan revealed 

significant linkage to markers on bovine chromosome 5 for the 

mutation causing arachnomelia or spider legs in Swiss and Italian 

Brown cattle. To select against these genetic defects either linked 

microsatellite markers (Weaver, SDM and arachnomelia) or the 

determination of the causative mutation itself (SMA) can be used to 

identify carriers. These molecular findings allow efficient selection 

against the four most frequently genetic diseases in Brown cattle. 

336 Bovine Leucocyte Adhesion Deficiency: History and Control. 

A Retrospective Survey 

H. Nagahata 

Rakuno Gakuen University, Department of Animal Health, Rakuno, 

Japan 

Bovine leucocyte adhesion deficiency (BLAD) in Holstein cattle is 

an autosomal recessive congenital disease characterized by recurrent 

bacterial infections, delayed wound healing and stunted growth, and 

is also associated with persistent marked neutrophilia. The 

molecular basis of BLAD is a single point mutation (adenine to 

guanine) at position 383 of the CD18 gene, which caused an 

asparatic acid to glycine substitution at amino acid 128 (D128G) in 

the adhesion molecule CD18. Neutrophils from BLAD cattle have 

impaired expression of the beta-2 integrin (CD11a,b,c/CD18) of the 

leucocyte adhesion molecule. Abnormalities in a wide spectrum of 

adherence dependent functions of leucocytes have been 

characterized. Cattle affected with BLAD have severe ulcers on oral 

mucous membranes, severe periodontitis, loss of teeth, chronic 

pneumonia and recurrent or chronic diarrhea. Affected cattle die at 

an early age due to the infectious complications. Holstein bulls, 

including carrier sires that had a mutant BLAD gene in heterozygote 

were controlled from dairy cattle. The control of BLAD in Holstein 

cattle by publishing the genotypes and avoiding the mating between 

BLAD carriers was found to be successful. The overview of the 

BLAD in Holstein cattle will be presented. 

365 Spastic Paresis and Spastic Syndrome: a Never Ending Story 

A. Gentile 


Italy 

Despite the fact that Spastic Paresis and Spastic Syndrome have been 

known for many decades, many of their pathogenic and etiological 

mechanisms remain unknown. Spastic Paresis is clinically 

characterized by a hyperextension of the rear limbs (“straight hock”) 

due to a contraction of the muscles which form the Achilles tendon. 

Signs of Spastic Paresis appear usually at the age of 3 to 8 months. 

Affected animal has a stiff gait and moves without normal hock 

flexion. In the most severe cases the leg is held so that the foot just 

touches the ground with the toe or remains completely raised from 

the ground and extended backwards. Spastic Paresis is considered to 

be caused by a spastic contraction of the antigravitational, foot 

extensor muscle group, especially the gastrocnemius and the 

superficial digital flexor. The contraction of the quadriceps femoris 

characterizes the recently described atypical form of Spastic Paresis 

of the femoral quadriceps, observed firstly in Belgian White Blue 

calves but experienced also in Romagnola animals. The muscular 

spasticity should be attributed to a hyperactivity of the myotatic 

reflex (“stretch reflex”). The dysfunction lies primarily in the 

gamma-pathway. Spastic Syndrome (“crampiness”, “Krämpfigkeit”) 

is a chronic condition that occurs in adult cattle. It is a particular 

problem in mature bulls maintained in artificial insemination centres 

but can affect also cows in a recurrent or in a progressive form. The 

disease is characterized by intermittent bilateral spasm of skeletal 

muscles of the pelvic girdle, including the muscles of the rump. Each 

Genetics Diseases Workshop 331

spasm is accompanied by kyphosis, which is often terminated by 

tremor of the hindquarters. During the attack one hind leg may be lift 

laterally in partial flexion. The literature is lacking in complete 

research on Spastic Syndrome and the few reports which included 

morphological examinations failed to find significant outcomes and 

concluded that the syndrome is a type of idiopathic or true muscle 

cramps. 

366 Genetic Diseases of the Dexter Breed: Chondrodysplasia and 

Hydrops Fetalis with Pulmonary Hypoplasia 

I. Tammen, J. Cavanagh, P. Windsor, F. Nicholas, H. Raadsma 

Faculty of Veterinary Science - University of Sydney, ReproGen, 

Sydney, Australia 

A form of dwarfism, commonly known as bulldog dwarfism, has been 

reported in Dexter cattle since the 19th century and presents one of the 

earliest single-locus gene disorders described in animals. Affected 

foetuses display extreme disproportionate dwarfism due to 

chondrodysplasia and are aborted or die at birth, whereas heterozygotes 

present with a milder form of disproportionate dwarfism - the “shortlegged” 

phenotype. We have identified two mutations cosegregating 

with the disease in the bovine aggrecan gene (ACAN): a common 4-bp 

insertion in exon 11 and a rarer transition in exon 1. Commercial DNA 

testing is now available to control this disorder. 

367 Genetic Diseases of Japanese Breeds 

T. Kunieda 

Okayama University, Graduate School of Natural Science and 

Technology, Okayama, Japan 

In the breeding of domestic animals, selection of economically desired 

traits has been the most important consideration for the improvement of 

animals, but exclusion of negative factors in animal production, such as 

genes for hereditary diseases, is also required for the genetic 

improvement of domestic animals. The incidences of many recessive 

hereditary diseases have been reported in Japanese beef cattle breeds 

and these diseases have caused serious problems for breeding of the 

beef cattle. These diseases include Chediak-Higashi syndrome, renal 

tubular dysplasia, chondrodysplastic dwarfism, factor XI deficiency, 

and congenital eye abnormality. We have identified the mutations 

responsible for these diseases by positional or functional cloning 

method and established the genotyping systems which can be used for 

identification of the carriers of these diseases. The allelic frequencies of 

the disease alleles and incidences of these diseases in the population of 

the cattle have remarkably reduced by application of these genotyping 

systems. 

368 Surveillance Programmes for Genetic Recessives: the 

Experience of Danish Bovine Genetic Disease Programme 

J. Agerholm 

University of Copenhagen, Faculty of Life Sciences, Department of 


The Danish bovine genetic disease programme was established in 

1989 by the breeding associations, the Faculty of Life Sciences and 

the Danish Veterinary and Food Administration. The aim was to 

establish a system that could handle suspected cases of 

inherited defects and to create a surveillance programme. The 

system was founded on research in pathology to ensure a high degree 

of diagnostic reliability and it should be able to handle cases 

independently of the breeding associations.The programme was 

initially faced by problems created by spinal muscular atrophy 

(SMA) in Danish Red cattle. The prevalence of SMA seemed to be 

very high. The first step was to differentiate between SMA and other 

disorders, which automatically led to the second step of identifying 

heterozygous sires. Once heterozygous males had been identified, 

these could be taken out of service thus reducing the prevalence of 

SMA. During the surveillance of SMA, cases were submitted with 

other clinical signs, i.e. congenital recumbency. Histopathology 

demonstrated myelin defects in the spinal cord, while genealogic 

examinations showed a familial pattern. The lesions corresponded to 

Spinal Dysmyelination, a disorder already identified by German 

researchers. The prevalence was reduced during the following years 

by identification and culling of carriers.The bovine leukocyte 


adhesion deficiency (BLAD) was identified in Denmark based on 

reports from USA and Japan. The disorder had remained undetected. 

The prevalence of BLAD was reduced to around zero within less that 

a year, as the molecular basis for the disorder was known and 

genotyping tests were available.The Complex Vertebral 

Malformation (CVM) syndrome was identified in Denmark in 1999. 

Identification of carriers was initially based on progeny 

examination, but was rapidly replaced by genotyping thus making a 

rapid decrease in prevalence possible. The strength of the Danish 

surveillance programme is the solid core of researchers dealing with 

inherited disorders, the awareness among veterinarians and cattle 

advisers and the high level of pedigree registrations in Danish cattle. 

The weakness of the system is the farmers’ reluctance to report 

malformed calves while still available for examination and the 

difficulties associated with dispatching cases for laboratory 

examination. The surveillance focuses on disorders detectable by the 

breeder or veterinarian, mostly disorders in the skin, skeletal or 

neuromuscular systems. 

369 Surveillance Programmes for Genetic Recessives: the 

Experience of the French Bovine Genetic Disease 

Observatory 

A. Eggen 1 , L. Manciaux 2 , S. Floriot 1 , A. Malafosse 3 , A. Ducos 4 

1 

INRA, Laboratoire de Génétique Biochimique et de Cytogénétique, 

Jouy en Josas, France 

2 

CEIA du Doubts et du Territoire de Belfort, Laboraoire de 

Génétique Biochimique et de Cytogénétique, Roulans, France 

3 

UNCEIA, PARIS, France 

4 

INRA, Station d'Amelioration Génétique des Animaux, Centre de 

Toulouse, Castanet-Tolosan, France 

The so called “French Bovine Genetic Disease Observatory” aims at an 

early detection of hereditary defects in cattle populations. It was built 

up in 2002 by the INRA Department of Animal Genetics and all 

representative professional organizations involved in cattle production 

and breeding. A standardized sheet allowing the declaration and 

description of congenital defects has been widely diffused. More than 

2000 individual declarations have been received to date, i.e. between 

350 and 550 per year. Eighty percent of these declarations concerned 

the Holstein Friesian breed. Only two particular defects were 

recurrently reported. 

370 The World-database of the Genetic Diseases of the Brown 

Breed 

L. Casanova 

European Brown Swiss Federation, Zug, Switzerland 

Genetic defects can only be fought by means of breeding, if they are 

recognized and announced in the field. The diagnosis of the defect as 

well as the ancestry of the affected animals are to be confirmed by 

suitable laboratory procedures. Recognized carriers of genetic defects 

have to be marked afterwards. This assures the information of the 

breeders. Since bulls are marketed world-wide today, it is 

indispensable that the information on genetic defects is interlaced 

internationally. In the year 2002, the European Brown Swiss 

Federation decided to develop an international data base for genetic 

defects in the Brown Swiss breed. This data base is operated by the 

Swiss Brown Cattle Breeders' Federation and contains the current 

information on the genetic defect status of male animals. 

410 The Point of View of the Geneticists: Genetic Analysis of 

Congenital Anomalies in Cattle in the Age of the Genome 

Sequence 



In the past the molecular genetic analysis of congenital anomalies in 

cattle was hindered by limited knowledge of the bovine genome. 

Therefore there were only few examples of successful molecular 

exploration of particularly monogenic inherited bovine disorders. 

With the decision to sequence the bovine genome project the 

situation changed in the year 2005 and today we have more or less 

excellent genomic resources for studying genetic conditions in 

cattle. Recently an appropriate mapping tool (SNP chip) for whole

genome association mapping in cattle became available. These 

outstanding bovine genome resources together with the well 

characterised relationship between the human and cattle genomes 

currently facilitate analyses of well known simple diseases as well as 

more complex inherited traits in cattle. 

411 The Point of View of the Geneticists: Online-based Approach 

to the Genetic Diseases of Cattle 

I. Tammen, F. Nicholas 

Faculty of Veterinary Science, University of Sydney, ReproGen, 

Sydney, Australia 

The increasing information on inherited diseases in animals can make 

it difficult for veterinarians, animal breeders and scientist to keep up to 

date with emerging inherited disorders. Online Mendelian Inheritance 

in Animals (OMIA: http://omia.angis.org.au/) is a database created by 

one of us (FWN) to summarise all the available information on 

inherited disorders and traits in more than 135 animal species (other 

than human and mouse, which have their own resources). There are 

currently 369 entries for cattle, including 37 disorders for which a 

molecular explanation (and hence DNA testing) is available. The 

database contains textual information and references, as well as links to 

relevant PubMed and Gene records at the National Center for 

Biotechology Information (NCBI), and is cross-referenced with the 

equivalent human database OMIM. By providing remote curator 

access, the database provides opportunities for experts in the field to 

enter additional information. 

412 Approach to the Genetic Diseases of a Service Company: 

Swissgenetics 

A. Pozzatti 

Swissgenetics, Zollikofen, Switzerland 

The following points summarize the monitoring system and the 

procedures carried out at different levels by the Swissgenetics in order 

to avoid the spreading of recessives in cattle population improved by 

its sires. 

Breeding Organizations: 

- Survey and analysis of information collected from progeny testing 

- Observing the frequency of hereditary defects in the different breeds 

and reporting 

- Keeping a list of the known hereditary defects and establishing a 

chart of the carrier-bulls for each breed Business 

Process Genetics: 

- Collecting data from the Breeding Associations 

- Checking the male veal calves, before purchase, for hereditary 

defects, as far as a test is available 

- Keeping a list of the known hereditary defects and establishing a 

chart of the hereditary defects of Swissgenetics bulls 

- Applying for measures 

- Converting the measures Business 

Process Production: 

- Clinical and eventually cytogenetic exam of the bulls 

- Technical support in keeping the list of hereditary defects 

- Up-dating of the documents 

413 The so Called Paunch Calf Syndrome in Romagnola Calves 

A. Gentile 1 , M. Rossi 1 , C. Droegemueller 2 

1 


Italy 

2 


“Paunch Calf” is the term used to indicate a congenital defect 

observed recently in Romagnola and Chianina calves.Calves show 

enlarged and floating abdomen and facial deformities characterized 

by shortened and flattened face and in some cases by enlarged head. 

A disproportionate shortness of the limbs (rhizomelia) is seldom 

observed. More frequent is the presence of cleft palate and the lack 

of an accessory hoof. At necropsy all animals show marked 

subcutaneous oedema, especially in the ventral part of the abdominal 

wall. Different quantities of ascitic fluid are constantly present in the 

abdominal cavity. Trasudate in pleural and pericardial cavity can 

also be present. Another characterizing aspect is the moderate to 

severe diffuse hepatic fibrosis, associated to the presence of hepatic 

cysts, containing serous or reddish fluid.Microscopic examination of 

the liver reveals an extensive distortion of lobular architecture with 

widespread fibrosis in periportal areas and around centrolobular 

veins. In some lobules the fibrosis is extended to perisinusoidal 

spaces. Cardiac malformations (atrial and/or interventricular septal 

defect, patent ductus arteriosus) constitute frequent accompanying 

findings. 

414 Congenital Muscular Dystony in the Belgian Blue Breed 

F. Rollin, S. Vandeputte, H. Guyot, K. Touati, D. Desmecht, 

C. Charlier, M. Georges 

University of Liege - Faculty of Veterinary Medicine, Liege, Belgium 

Over the last decades, two different forms of Congenital Muscular 

Dystony (CMD) were observed with an increasing incidence (0.1- 

0.2%) in Belgian Blue (BB) calves. CMD1 calves present since birth 

various degrees of dysphagia and locomotor troubles. In most cases, 

they are unable to get up without help. Once standing up, some 

hesitate to move in a voluntary way and are quickly tired whereas 

others walk with more ease. When they are stimulated or pushed, 

they become stiff and fall without flexing the legs, like a plank. 

When suckling, they show swallowing difficulties associated with a 

stertorous cough and tachypnoea. They usually die within a few 

weeks as a result of recumbency and respiratory complications, but a 

few calves adapt to their handicap and survive in spite of obvious 

growth retardation. The first symptoms of CMD2 are generally 

detected on the occasion of the C-section, when the calf still is in the 

uterus. They consist of severe episodes of myoclonus of long 

duration triggered off by tactile or acoustic stimulation. These calves 

die within a few hours of birth if fits of apnea are not prevented by 

IM diazepam administration. Clinically, CMD2 is clearly 

reminiscent of congenital myoclonus described in Hereford cattle. 

Genealogical analysis suggested that both CMD1 and 2 were 

recessive inherited defects. Recently, the genes and causative 

mutations of these defects were located on the chromosome BTA25 

and 29, respectively for the CMD1 and 2 (Charlier et al., 2008: 

http://www.nature.com/ naturegenetics).For CMD1, DNA 

sequencing revealed a missense mutation (C1676T) leading to a 

R559C substitution in exon 14 of ATP2A1, which encodes a fasttwitch 

skeletal-muscle Ca++ ATPase. ATP 2A1 (also known as 

SERCA1) is responsible for pumping Ca++ from the cytosol back 

into the sarcoplasmic reticulum, thereby inducing muscle relaxation. 

The mutation regarding CMD2 concerns SLC6A5 that encodes the 

Na+/Cl- dependent glycine transporter (GlyT2), so disrupting 

presynaptic uptake of glycine by GlyT2. The corresponding human 

disorders for CMD1 and 2 are Brody myopathy and hyperekplexia, 

respectively. All BB A.I. sires were genotyped for both defects and 

it was found that 11.4% were carriers of CMD1, 4.6% of CMD2 and 

0.6% of both. 

415 Congenital Pseudo-myotonia in Chianina Cattle 

S. Testoni 1 , A. Gentile 2 , P. Boni 3 

1 

University of Padova, Department of Veterinary Clinical Sciences, 

Padova, Italy 

2 

University of Bologna, Department of Veterinary Clinical Sciences, 

Bologna, Italy 

3 

Bovine Practitioner, Bovine Practitioner, Perugia, Italy 

The authors present a new muscle function disorder observed in 

genetically related Chianina animals. The main clinical feature is an 

exercise-induced muscle contracture. When animals are stimulated 

to move faster, the muscles immediately become stiff and temporary 

“freeze up”, inducing a stiff gait. Scuffing the toe and bunny hopping 

are other progressive exercise-induced findings in the hind legs, 

whereas goose-stepping and circumduction (swinging outwards) are 

additional alterations in the front legs. Animals also experience 

general muscular stiffening when startled, associated with a 

retraction of the bulbi and prominence of the third eyelid. During the 

startle-elicited episodes consciousness is preserved. At a slow pace, 

the animals do not show any stiffness, even when walking for a long 

time; no sign of weakness is noted. Symptoms are evident since birth 

and remain practically unchanged for life. Biochemistry usually 

shows slightly increased levels of CK, LDH and AST. L-Lactate is 

practically normal at rest and after a crisis-free slow paced walk, but 

increases immediately after stronger exercise accompanied by the 

Genetics Diseases Workshop 333

occurrence of a crisis of stiffness. EMG investigations at rest and 

during exercise do not show any pathological findings. In the 

experience of the authors, histopathological examination of the 

semimembranosus muscle show signs of rhabdomyolysis; dark giant 

fibers with large areas of necrosis can be observed in type 2 muscle 

fibers. Metabolic muscular disturbances are excluded by 

hystochemical techniques, such as SDH associated to COX. Using 

the anti-neonatal antibody it is possible to observe regenerating 

fibers in the periphery of the rhabdomyolytic areas. On the basis of 

the clinical findings and of the results obtained by 

electromyographic and histological examinations, the authors 

suggest the term “Congenital Pseudo-Myotonia”: as opposed to true 

myotonia, in fact, the electromyographic examination does not show 

any alteration, not even during the phase of the muscle contraction. 

Similarly to the so-called “Brody's disease” of the human medicine, 

there are evidences that attribute the condition to an exerciseinduced 

impairment of muscle relaxation due to a defect of calcium 

reuptake. Immunoblotting analysis (carried out in two affected 

animals) indicates a remarkable reduction in SERCA1 (Sarco- 

Endoplasmic Reticulum Ca2+ATPase 1) content in muscles. 

416 Congenital Erythropoietic Porphyria in Holstein Cattle 

J. Agerholm 1 , C. Jorgensen 1 , C. Bendixen 2 , M. Fredholm 1 

1 Faculty of Life Sciences, University of Copenhagen, Department of 


2 Faculty of Agricultural Sciences, University of Aarhus, Department 

of Genetics and Biotechnology, Tjele, Denmark 

Objectives of study: Congenital erythropoietic porphyria (CEP) is 

an autosomal recessive inborn error of haem biosynthesis and is 

associated with accumulation of porphyrins in several tissues. It is a 

severe disease in Holstein cattle leading to photosensitization and 

subsequent necrosis of unpigmented areas of the skin due to the 

photodynamic properties of porphyrins. It is important to restrict the 

prevalence of this disorder in cattle, i.e. by culling of carriers. 

Mutations in the uroporphyrinogen III synthase gene (UROS) cause 

CEP in man. Direct measurements of UROS activity in cattle have 

shown a much lower activity in affected animals than in non-affected 

animals making UROS a candidate gene. Taking the outset in these 

observations a study was performed to identify the molecular cause 

of CEP in Danish Holsteins. 

Materials and Methods: Five cases of CEP observed during a period 

of 18 years were included. Muscle tissue was available from two cases 

and DNA was available from 13 animals related to affected animals. 

Blood samples were collected from 10 unrelated individuals of the 

Holstein breed. DNA was extracted from the muscle and blood 

samples.Based on sequence information from the assembled bovine 

genome, primers were designed for sequencing the exons of the bovine 

UROS gene. Sequence information was generated from the two 

affected individuals and the unrelated individuals. The sequences were 

aligned and manually curated for SNPs. 

Results: Pedigree analyses demonstrated that the five cases were 

genetically related to a common ancestor and occurred in a pattern 

consistent with autosomal recessive inheritance.Sequencing of the 

UROS exons including flanking regions resulted in the identification 

of three intronic SNPs. Both affected individuals were homozygous 

for two of these SNPs, which were not identified in any of the 

unrelated non-affected individuals. The segregation of the SNPs will 

be analysed in putative heterozygous animals to confirm that a 

mutation in UROS is the underlying cause of CEP in Danish 

Holsteins. 

Conclusions: CEP occurs in Danish Holstein as an autosomal 

recessively inherited defect. The disorder is most likely caused by a 

mutation in UROS. 

417 Crooked Tail and Muscular Hypertrophy Syndrome, an 

Emerging Inherited Defect in Belgian Blue Breed 


A. Sartelet 1 , M. Dive 2 , K. Touati 1 , F. Rollin 1 , M. Georges 3 , 

C. Charlier 3 

1 

University of Liege, Clinical department of food animals, Liege, 

Belgium 

2 

Practitioner, Ciney, Belgium 

3 

University of Liege, Unit of Animal Genomics, GIGA-R & Faculty of 

Veterinary Medicine, Liege, Belgium 

Intense selection for desired production traits in livestock often 

results in an increase of the rate of inbreeding. The use of artificial 

insemination, combined with a restricted number of fashionable 

sires, reinforces the problem. This causes frequent emergence of 

inherited defects which can affect such a high proportion of the 

population that they become a major economic problem. About one 

year ago, cattle practitioners became very concerned by an emerging 

defect in Belgian Blue cattle, a breed well known for its extreme 

muscularity. In many farms, three to four affected calves were born 

by hundred births. Such a high incidence forced us to perform a 

detailed clinical study to characterize the defect and try to determine 

its precise aetiology. In a matter of months, fifty affected calves were 

clinically examined, blood samples collected and their pedigrees 

scrutinized. Pedigree analysis was in agreement with an autosomal 

recessive mode of inheritance of the defect. The main clinical signs 

were extreme muscularity, growth retardation, postural abnormality, 

shortened head and crooked tail, allowing us to name the syndrome 

as “crooked tail and muscular hypertrophy syndrome” (CTMHS). In 

addition, a few cases exhibited scoliosis, joint rigidity, and spastic 

paresis of the hind limbs. Expression of these last signs was variable. 

Even if the defect was not lethal, euthanasia was sometimes 

necessary and economic loss due to growth retardation and clinical 

complications was important. Genome scans were performed using 

DNA samples from affected individuals and controls using a custom 

made 60K Illumina SNP (Single Nucleotide Polymorphism) 

panel. A unique identical-by-descent (IBD) chromosomal segment 

of 2.42 Mb, shared by all affected individuals, was mapped on 

bovine chromosome 19, proving the hereditary aetiology of this new 

syndrome. Candidate genes in this interval are currently analyzed in 

order to identify the causative mutation to be able to develop a 

diagnostic test. This will allow immediate eradication of the defect 

from the Belgian blue population and avoid the associated economic 

losses. 

Key words: cattle, Belgian Blue, emergence, inherited defect 

418 Ichthyosis Fetalis: an Emerging Lethal Condition 

Endangering the Belgian White and Red Cattle Breed 

P. Cornillie 1 , S. Cools 2 , L. Vandaele 2 , A. De Kruif 2 , P. Simoens 1 

1 Ghent University, Department of Morphology, Merelbeke, Belgium 

2 Ghent University, Department of Reproduction, Obstetrics and Herd 

Health, Merelbeke, Belgium 

In the last few years, several cases of ichthyosis fetalis have been 

reported in the Belgian White and Red cattle breed. This congenital 

skin disorder is characterised by the presence of thick, hyperkeratotic 

scaly plates separated by deep reddened fissures all over the body of the 

affected calves which usually die soon after delivery. In most cases, 

alopecia, ectropion and eclabium are additional typical findings. Based 

on pedigree analysis, autosomal recessive inheritance of this condition 

is suggested. By analogy with human harlequin ichthyosis, a disorder 

morphologically greatly resembling bovine ichthyosis fetalis, it is 

highly suspected that a defective lipid transport in keratinocytes caused 

by severe mutations at the level of the ABCA12 gene is involved in the 

pathomechanism of the disorder. Emergence of this condition in a 

population that is already compromised by inbreeding seriously affects 

the prospects for the preservation of this relatively small and regionally 

confined herd, especially as some bulls that are most commonly used 

for breeding purposes have been identified as carriers of the defective 

allele.

Notices 

Notices 335

Notices 

336 XXV. Jubilee World Buiatrics Congress 2008

World Association for Buiatrics 

PRESIDENT 

W. Baumgartner, Austria 

VICE-PRESIDENTS 

R. Ugarte, Uruguay 

B. Rheinberger, Australia 

SECRETARY GENERAL 

O. Szenci, Hungary 

MEMBERS OF THE COMMITTEE 

E. Bouchard, Canada 

J. Cannas Da Silva, Portugal 

R.G. Eddy, United Kingdom 

A. El-Sebaie, Egypt 

M. Garcia, Brasil 

H. R. Han, South Korea 

N. Jessen, Denmark 

T. Lam, the Netherlands 

K. Müller, Germany 

T. Nakao, Japan 

H. Navetat, France 

OBSERVER 

M. G. Riddell, USA 

HONORARY PRESIDENTS 

H. E. Amstutz, USA 

H. J. Breukuink, the Netherlands 

P. D. Videla, Argentina 

HONORARY VICE-PRESIDENT 

R. Dubois, Brasil 

HONORARY SECRETARY 

P. Stöber, Germany 

P. Lekeux, Belgium 

HONORARY MEMBERS 

C. G. Dirksen, Germany 

K. Hamana, Japan 

A. D. Weaver, United Kingdom 

W. Hofmann, Germany 

Hungarian Association for Buiatrics 

PRESIDENT 

Prof. Dr. Szenci Ottó 

SECRETARY 

Prof. Dr. Brydl Endre 

MEMBERS 

Dr. Abonyi Tamás 

Dr. Bartyik János 

Gulyás Tibor 

Dr. Kálmán Imre 

Dr. Kocsis László 

Dr. Lehoczky János 

Dr. Mészáros Gyula 

Sztakó István 

Dr. Sümeghy László 

HONORARY MEMBERS 

Prof. Dr. Dr. h.c Kovács Ferenc 

Prof. Dr. Dr. h.c. Németh Ferenc 

Dr. Vajdovich Károly 

List of the Biennale World Buiatrics 

Congresses 

Hannover (Germany) 1960 

Vienna (Austria) 1962 

Copenhagen (Denmark) 1964 

Zurich (Switzerland) 1966 

Opatija (Yugoslavia) 1968 

London (UK) 1970 

Philadelphia PA (USA) 1972 

Milan (Italy) 1974 

Paris (France) 1976 

Mexico City (Mexico) 1978 

Tel Aviv (Israel) 1980 

Amsterdam (The Netherlands) 1982 

Durban (South Africa) 1984 

Dublin (Ireland) 1986 

Palma de Mallorca (Spain) 1988 

San Salvador da Bahia (Brazil) 1990 

St Paul MN (USA) 1992 

Bologna (Italy) 1994 

Edinburgh (UK) 1996 

Sydney (Australia) 1998 

Punta del Este (Uruguay) 2000 

Hannover (Germany) 2002 

Quebec City (Canada) 2004 

Nice (France) 2006

Oral and Poster Abstracts

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