Biotechnology in Animal Husbandry - Institut za Stočarstvo

UDC636 Print ISSN 1450-9156
Online ISSN 2217-7140
VOL 28, 3
Founder and publisher
INSTITUTE FOR
ANIMAL HUSBANDRY
11080 Belgrade-Zemun
Belgrade 2012
BIOTECHNOLOGY
IN ANIMAL HUSBANDRY
CONTENTS
Z. Pavlovski, Z. Škrbiæ, M. Lukiæ, D. Vitoroviæ, S. Liliæ, V. Petrièeviæ
SHELL QUALITY – EVERLASTING PROBLEM IN THE TODAY POULTRY
SCIENCE...................................................................................................................
N. Mincheva, M. Lalev, M. Oblakova, P. Hristakieva, I. Ivanova
EFFECT OF FEATHERING ALLELES (K/K + ) ON LAYING PERFORMANCE,
HATCHABILITY PARAMETERS AND SOME BODY MEASUREMENTS IN
TWO LINES OF WHITE PLYMOUTH ROCK HENS............................................
N. Tolimir, L. Periæ, N. Miloševiæ, M. Ðukiæ – Stojèiæ, R. Jovanoviæ, V.
Bogdanoviæ
THE EFFECT OF PHASE NUTRITION DURING STARTER PERIOD ON
PRODUCTION PERFORMANCES AND NITROGEN CONTENT IN FECES OF
BROILERS OF DIFFERENT GENOTYPES...........................................................
M. Ðukiæ Stojèiæ, N. Miloševiæ, L. Periæ, I. Jajiæ, N. Tolimir
EGG QUALITY OF JAPANESE QUAIL IN SERBIA (Coturnix coturnix japonica)
E.N.Uchewa,P.N.Onu..........................................................................................
THE EFFECT OF FEED WETTING AND FERMENTATION ON THE
PERFORMANCE OF BROILER CHICK.......................................................................
M. Sahraei, H. Janmmohamadi, A. Taghizadeh, G. Ali Moghadam, S. A. Rafat
ESTIMATION OF THE RELATIVE BIOAVAILABILITY OF SEVERAL ZINC
SOURCES FOR BROILERS WHEN FED A CONVENTIONAL DIET.................
V. Vidoviæ, D. Lukaè, M. Stupar, V. Višnjiæ, J. Krnjaiæ
HERITABILITY AND REPEATABILITY ESTIMATES OF REPRODUCTION
TRAITS IN PUREBRED PIGS.................................................................................
È. Radoviæ, M. Petroviæ, N. Parunoviæ, N. Brkiæ, B. �ivkoviæ, M. Gogiæ, N.
Stanišiæ
THE EFFECT OF GENOTYPE AND YEAR ON TRAITS OF PERFORMANCE
TESTED GILTS.........................................................................................................
M. Popovac, D. Radojkoviæ, M. Petroviæ, M. Mijatoviæ, M. Gogiæ, D. Stanojeviæ,
N. Stanišiæ
HERITABILITY AND CONNECTIONS OF SOW FERTILITY TRAITS.............
Ð. Okanoviæ, D. Ivanov, D. Paliæ, A. Mandiæ, N. Iliæ
MEAT FATTY ACID PROFILE OF PIGS FED LINSEED ENRICHED DIET......
R. Ðedoviæ , V. Bogdanoviæ, G. Trifunoviæ, M. Petroviæ, M. M. Petroviæ, D.
Stanojeviæ
THE EFFECT OF THE LEVEL OF MILK YIELD ON THE REPRODUCTION
TRAITS IN BLACK AND WHITE COWS..............................................................
D. Nikšiæ, V. Panteliæ, D. Ostojiæ-Andriæ, P. Perišiæ, M. Petrièeviæ, R. Ðedoviæ, M.
Lazareviæ
RESULTS OF THE BIOLOGICAL TEST OF SIMMENTAL BULLS IN
CENTRAL SERBIA..................................................................................................
R.T. Gudaj, E. Brydl, J. Lehoczky, I. Komlósi
STUDY OF ANIMAL WELFARE STATUS IN DAIRY COW HERDS IN
HUNGARY................................................................................................................
M.P.Petroviæ, V. Caro Petroviæ, D. Ru�iæ Musliæ, Z. Ilic, B. Miloseviæ,, J.Stojkoviæ,
N. Maksimoviæ
SOME IMPORTANT FACTORS AFFECTING FERTILITY IN SHEEP................
D.Ru�iæ-Musliæ,M.P.Petroviæ,M.M.Petroviæ,Z.Bijeliæ,V.Panteliæ,P.Perišiæ,
V. Caro-Petroviæ
THE EFFECT OF THE SYSTEM OF CROSSING ON FATTENING
PARAMETERS OF WEANED LAMBS.................................................................
393
405
415
425
433
441
455
463
469
477
487
497
509
517
529

Z. Iliæ, J. Stojkoviæ, D. Ru�iæ Musliæ, V. Caro Petroviæ, M. P. Petroviæ, R. Djokoviæ,
V. S. Kurèubiæ
THE INFLUENCE OF BIOLOGICALLY ACTIVE SUPPLEMENT "BIORIL"ON
PERFORMANCE OF FATTENING LAMBS............................................................
J. Stojkoviæ, Z. Iliæ, S. Æiriæ, B. Ristanoviæ, M. P. Petroviæ, V. Caro Petroviæ, V.
Kuræubiæ
EFFICENCY OF ZEOLITE BASIS PREPARATION IN FATTENING LAMBS
DIET.............................................................................................................................
M. Joksimoviæ Todoroviæ, V. Davidoviæ, Lj. Sretenoviæ
THE EFFECT OF DIET SELENIUM SUPPLEMENT ON MEAT QUALITY.........
D. Trboviæ, D. Vraniæ, J. Djinovic-Stojanoviæ, V. Matekalo-Sverak, V. Djordjeviæ,
J. Babiæ, D. Spiriæ, R. Petronijeviæ, A Spiriæ
FATTY ACID PROFILE IN RAINBOW TROUT (ONCORHYNCHUS MYKISS)
AS INFLUENCED BY DIET......................................................................................
H. F. Abou-Shaara, A. A. Al-Ghamdi
STUDIES ON wINGS SYMMETRY AND HONEY BEE RACES
DISCRIMINATION BY USING STANDARD AND GEOMETRIC
MORPHOMETRICs....................................................................................................
S. Jankoviæ, S. Rakiæ, , J. Ikanoviæ, J. Kuzevski, Lj. �ivanoviæ, �. Lakiæ
CORRELATION COEFFICIENTS OF MORPHOLOGICAL-PRODUCTIVE
TRAITS OF SPECIES OF SORGHUM GENUS ......................................................
S. Rakiæ, S. Jankoviæ, M. Demin , D. Bucalo, M. Maslovariæ
QUALITY AND CONDITION OF WHEAT GRAIN (Triticum spp.) DURING
STORAGE....................................................................................................................
A. Akinfemi
UPGRADING OF SUGARCANE BAGASE BY SOLID STATE
FERMENTATION WITH PLEUROTUS SAJOR-CAJU AND PLEUROTUS
FLORIDA AND THE IMPACT ON THE CHEMICAL COMPOSITION AND IN
VITRO DIGESTIBILITY............................................................................................
Ya. Kozhouharov, V. Lingorski
INFLUENCE OF MINERAL FERTILIZATION ON SOME BIOLOGICAL AND
PRODUCTIVE INDICATORS OF NATURAL MEADOW OF AGROSTIS
CAPILLARIS-FESTUCA FALLAX TYPE IN THE RHODOPE MOUNTAINS
(SOUTHERN BULGARIA).......................................................................................
S. Ðedovic, M. Vukša , M. M. Petroviæ, J. Bojkovski, I. Pavloviæ, G. Jokiæ, B.
Stojniæ
CONTROL OF BROWN RAT (Rattus norvegicus) ON A DAIRY FARM IN
SERBIA........................................................................................................................
537
545
553
563
575
585
595
603
613
623

Journal for the Improvement of Animal Husbandry
UDC636 Print ISSN 1450-9156
Online ISSN 2217-7140
BIOTECHNOLOGY
IN ANIMAL HUSBANDRY
Belgrade - Zemun 2012

Biotechnology in Animal Husbandry 28 (3), p 393-633, 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636
Editorial Council
Prof. Dr Milica Petrović, president
Prof. Dr Lidija Perić, full prof.
Prof. Dr Vojislav Pavlović, full prof.
Dr. Zoran Lugić, science advisor
Editor’ s Office
Prof. Dr. Martin Wähner, Germany
Dr. Branislav Živković, Serbia
Dr. Marin Todorov, Bulgaria
Dr. Milan M. Petrović, Serbia
Prof. Dr. Kazutaka Umetsu, Japan
Prof. Dr. Dragan Glamočić, Serbia
Prof. Dr. Vigilijus Jukna, Lithuania
Dr. Elena Kistanova, Bulgaria
Prof. Dr Zlatko Skalicki, full prof.
Dr Miroslav Blagojević
Dr Branka Vidić, science advisor
Prof. Dr. Wladyslaw Migdal, Poland
Prof. Dr. Colin Whitehead, United Kingdom
Dr. Branislav Bobček, Slovak Republic
Prof. Dr. Sandra Edwards, United Kingdom
Dr. Vojislav Mihailović, Serbia
Prof. Dr. Giacomo Biagi, Italy
Prof. Dr. Stelios Deligeorgis, Greece
Prof. Dr. Hasan Ulker, Turkey
Dr. Catalin Dragomir, Romania
On behalf of publisher
Miloš Lukić, PhD, Research Fellow, Director of the Institute for Animal Husbandry, Belgrade-Zemun, Serbia
Editor in Chief
Zlatica Pavlovski, PhD, Science Advisor, Institute for Animal Husbandry, Belgrade-Zemun, Serbia
Deputy Editor in Chief
Zorica Tomić, PhD, Science Advisor, Institute for Animal Husbandry, Belgrade-Zemun, Serbia
Editor
Miloš Lukić, Ph.D, Research Fellow, Institute for Animal Husbandry, Belgrade-Zemun, Serbia
Section Editors
Genetics and breeding
Milan P. Petrović, Ph.D, science advisor
Reproduction and management
Miroslav Žujović, Ph.D, science advisor
Nutrition and physiology of domestic animals
Ljljana Sretenović, Ph.D, science advisor
Language editor
Olga Devečerski, grad. prof.
Food safety, technology and quality of animal
products
Stevica Aleksić, Ph.D, science advisor
Sustainability of feed production and ecology
Zorica Bijelić, Ph.D, research fellow
Alternative production in livestock
Zdenka Škrbić, Ph.D, senior research fellow
Address of the Editor’s office
Institute for Animal Husbandry, Autoput 16, P. Box 23, 11080 Belgrade-Zemun, Republic of Serbia
Tel. 381 11 2691 611, 2670 121; Fax 381 11 2670 164; e-mail: biotechnology.izs@gmail.com; www.istocar.bg.ac.rs
Biotechnology in Animal Husbandry is covered by Agricultural Information Services (AGRIS) -Bibliographic
coverage of abstracts; Electronic Journal Access Project by Colorado Altiance Research Libraries -Colorado,
Denver; USA; Matica Srpska Library -Referal Center; National Library of Serbia; University Library "Svetozar
Markovic", Belgrade, Serbia
According to CEON bibliometrical analysis citation in SCI index 212, in ISI 9, impact factor (2 and 5) of
journal in 2007: 0,667 and 0,467, - M24 category
Annual subscription: for individuals -500 RSD, for organizations 1200 RSD, -foreign subscriptions 20 EUR. Bank
account Institut za stočarstvo, Beograd-Zemun 105-1073-11 Aik banka Niš Filijala Beograd.
Journal is published in four issues annually, circulation 100 copies.
The publication of this journal is sponsored by the Ministry of Education and Science of the Republic of Serbia.
Printed: "Mladost birošped", Novi Beograd, St. Bulevar AVNOJ-a 12, tel. 381 11 2601-506

Biotechnology in Animal Husbandry 28 (3), p, 393-404, 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 637.05
DOI: 10.2298/BAH1203393P
SHELL QUALITY – EVERLASTING PROBLEM IN THE
TODAY POULTRY SCIENCE
Z. Pavlovski 1 , Z. Škrbić 1 , M. Lukić 1 , D. Vitorović 2 , S. Lilić 3 ,
V. Petričević 1
1 Institute for Animal Husbandry, Autoput 16, P.Box 23, 11080 Belgrade, Republic of Serbia
2 Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade, Republic of Serbia
3 Institute of Meat Hygiene and Technology, Kaćanskog 13, Beograd, Republic of Serbia
Corresponding author: zlaticapav@yahoo.com
Review paper
Abstract: Shell breakage (6-20%) is of major concern for its economic
consequences and safety of egg consumption. A great deal of effort in the fields of
nutrition, genetics and enviromental conditions have been carried out to improve
eggshell quality but understanding the formation of the eggshell fabric and the
origin of shell defects is perhaps the key to solving eggshell problems. The aim of
this review is to summarise new information on eggshell problems. The aim of this
review is to summarise new infotmation on eggshell formation and update the
nutritional unfavourable enviromental conditions.
Keywords: Egg production, shell quality, problem, possible solutions
Introduction
The quality of the egg shell is still a major concern of many participants in
the poultry production chain, producers of table eggs and hatching egg producers.
The successful development of a chicken embryo depends on the quality of the egg
shell, i.e. its robustness for the purpose of protection, protection from infection and
water loss from the egg, and most importantly, it is a source of calcium for the
formation of the skeleton of the embryo (Lavelin et al., 2000). On the other hand,
in the commercial production and marketing, table eggs are exposed to many
shocks affecting percentage of broken and cracked eggs and thus directly affecting
the economic losses of the producer. In addition, there is growing concern about
the safety (hygiene compliance) of eggs, as the egg shell is the first barrier against
bacterial penetration of the egg and therefore must be free of defects. Ban on cage
system of housing hens in the EU, also increases the risk of infection from

394
Z. Pavlovski et al.
Salmonella, etc., so in that sense, the eggshell quality is very important (Nys,
2001).
Shell breakage is still a major concern of egg producers, because 80 to
90% of the eggs deficiencies relates to eggshell. Defects include eggs without shell
that are rarely observed and not counted in the number of fresh laid eggs, and the
eggs that acquired defects in body before oviposition which may be partially and
completely removed. The defective eggs include eggs irregular shape, eggs with
partially thin shell and eggs with a rugged shell (Bell, 1998). Shell eggs differ
depending on the chicken breed or hybrid, flock age, season, oviposition and
between chickens in the same flock.
Egg shell is a highly organized mineral structure built of spherical calcite
crystals deposited on the outer surface of the membrane around egg white. Crystals
of calcium carbonate (in the form of calcite) are formed by crystallization from
supersaturated solution of secretion from the distal fallopian tube - uterus. Layers
take about 20 hours to form a shell, which shows the huge requirements for
continuous supply of required quantities of calcium. Every interruption in calcium
supply to uterus leads to a decline in the quality of the shell and further economic
losses.
Harms et al. (1996) report that approximately 6-8% of the total egg production is
not usable/markatable due to poor quality of shells, and Roland (1988) points out
that 12-13% of eggs are lost on the way from the producer to the consumer for the
same reasons. The ability of eggshell to withstand the impact of the external force
is the strength of the shell (Hamilton, 1982). Many factors influence the quality of
the shell, and its strength, such as genetic basis, layer hen age, nutrition,
environmental conditions, disease, etc. (Washbourne, 1982; Vitorović et al., 1995,
Roberts and Nolan, 1997; Daghir 2004, Zhang et al., 2005, Pavlovski et al., 2012).
Eggs of better quality of egg shell are laid in the afternoon period compared to the
morning (Pavlovski and Vitorović, 1996; Škrbić et al., 1998; Tumova et al., 2007).
The main source of calcium in the hens diet is the finely ground limestone, chalk.
Shell formation takes place during the night and then layers consume less or not at
all food and calcium needs are not adequately met. The problem of the quality of
the egg shell over last 30 years, and it is still current, is subject of numerous studies
since broken egg shell depends not only on the strength of the shell, but also on
shell thickness and strength of "attack" on it.
In this paper, we focus on the formation of the egg shell, shell quality
problems and possible solutions for improving the quality of the shell.
Forming of the eggshell
Eggshell is formed in the uterus, expanded section of fallopian tubes called
the shell gland. After fertilization of the egg in the infundibulum and secretion of

Shell quality – everlasting problem in ...
egg white in the magnum, the egg enters the isthmus (narrowing of the tubes) 2 to
3 h after ovulation. In the isthmus, granular cells secrete various components of the
shell membranes such as type X collagen. In utero, from the supersaturated
solution of calcium carbonate (about 5 to 6 g) the crystals are formated that are
deposited in layers on the surface of the thin outer membrane protein. The quality
of the shell formed to a large extent depends on the size and shape of crystals on
the one hand and on the other on the interaction of calcium carbonate and organic
matrix.
A large number of matrix macromolecules have only recently been discovered and
identified. In the uterine fluid and organic shell secretions three types of proteins
have been discovered: egg white proteins (lysozyme, ovotransferrin, ovalbumin),
osteopontin, which was first identified in bone and uterine proteins that are unique
to the process of forming of shell: ovocleidin 17, 116; ovocalyxin 32 (Nys et al.,
2000). Egg white proteins are located in the membranes of the egg. They can
participate in the fight against microorganisms because of their microbial activity,
and lysozyme and ovotransferrin can affect the size and morphology of calcite
crystals when the crystals grow in vitro in the presence of these proteins. It is
assumed that greater protein participation in the eggshell affects the morphology of
the shell and the quality. It is known that the shell quality in older hens is worse
and shell breakage increased. The decline in shell thickness with the age of the
hens is related to the inability to reduce shell weight. Preliminary research indicates
that in the egg shell from older hens there are more matrix proteins. Further
research in this area should confirm the hypothesis that matrix proteins play a role
in the formation of the shell. Feeding hens remains of utmost importance, because
it is the only supplier of inorganic substances necessary for the formation of the
shell.
Eggshell defects
Common defects (deficiencies) of eggshell that may result in the economic
loss in the production of eggs (Egg Quality A Practical Approach, Roche) are:
large cracks, thin line fissures, rough cracks, eggs of deformed shape, equilateral
egg, eggs covered with cracks, pimply shells.
Measuring of the quality
Research literature abounds with research on the effects of nutrition, light
regime, population density, genetic bases and similar on the quality of the shell, but
what is the quality of the shell and how is it measured?
Of the physical properties of the eggshell the following traits are measured in order
to improve the quality of the shell:
395

396
Z. Pavlovski et al.
EGGSHELL WEIGHT, SHELL DEFORMATION, BREAKING FORCE
EGG LENGTH / WIDTH = EGG SHAPE INDEX
TOTAL SHELL THICKNESS
THICKNESS OF THE MAMMILARY LAYER
EFFECTIVE THICKNESS
FRACTURE TOUGHNESS
SPECIFIC WEIGHT
The right choice of the measure brings us to the real conclusion of the factors
influencing the quality of the eggshell and other aspects of the quality of the
product.
Possible solutions
Control of egg mass (shell). Egg mass is still of paramount importance for
the maintenance of good eggshell quality. Egg masses can be reduced by a lower
total amount of protein in the diet (19/15mg/day) or methionine levels (500/200 mg
/ day), but it can adversely affect the level of production. Supplementing 0.25 or
0.5% of amido ethyl-sulfonic acid in powder form for laying hens decreases the
egg mass and has no effect on production traits, but there is no information whether
there is any effect on the share of eggshell. The energy level can not be
manipulated, but there is abundant information in the literature describing the
influence of the fat on the egg mass. Adding 4% of fat affects the egg mass at the
start of production (Grobas, 1999), but at the end of production, adding fat should
be avoided, so the mass of the eggs would not be increased. Vegetable oils are rich
in unsaturated fatty acids and 0.81% linoleic acid (Harms et al., 2000) positively
affect egg mass. Using more saturated fatty acids (palm oil) and adding linoleic
acid in quantities that meet the daily needs of normal hens physiological function
(0.8%), are alternatives with which to control the increase in weight with age of
laying hens, especially when egg mass reduction does not reflect the change in
mass of the eggshell.
Calcium in the diet of chickens. In rearing chickens (8-16 weeks of age) the
calcium requirements range from 0.9 to 1,1%. Excessive doses of calcium in food
for longer period is not recommended because it can cause depression in the
growth of hens and kidney damage and increased mortality (Hamilton and Cipera,
1981). The most important period in the rearing is when the young hens become
laying hens. Calcium levels need to be increased two weeks before the laying starts
to facilitate the development of medullary bone, and more importantly to avoid that
hen lays the first egg without the addition of calcium in the diet that could result in
negative calcium balance that is difficult to correct later. Delays in the introduction
of a greater share of calcium has a negative effect on the quality of the egg shell of
the first laid egg and throughout the laying period has a negative impact on the

Shell quality – everlasting problem in ...
quality of the shell (Roland, 2000). Today, hens mature sexually much earlier and
therefore produce their first egg before they move into a facility for hens which
should coincide with the introduction of a meal for layers. Therefore, it is
recommended to add 2.5 to 3.5% calcium in the diet prior to beginning of laying,
that is, from 14 to 16 weeks of age of young hens.
Daily consumption of calcium by layers of is 2.2 g hence, the calcium
requirements are 4g. If the level of calcium in the diet is below 3%, it increases the
mortality and reduces the production of eggs, which has been confirmed in their
research by Keshavarz (1998 a, b) and Chandramoni et al. (1998). Optimal quality
of eggshell was determined in eggs from hens fed with a diet containing more than
3.5% calcium in layers of brown shell eggs (Vitorović et al., 1995; Safaa et al,
2008) and 4 to 4.5 g per day (Scott et al., 1999). Selection of hens for large number
of eggs is associated with a daily production of eggs, where more eggs are laid in
the morning or before the light is turned on (80% of the new-laid eggs 11 hours
subsequent to turning off of the lights, i.e. 3h after lights are turned on). Thus,
eating in the morning can not ensure a normal supply of calcium to form a shell
that is over 1.5 h before oviposition. Introducing the midnight feeding of hens
improves synchronization between calcium intake and the formation of shells and
shell quality, especially when you introduce the intermittent light (2 h) at night and
only when the line for feeding of hens is on.
The particle size of calcium. The main source of calcium in laying hens
feed is the finely ground limestone, chalk. Eggshell formation takes place mainly at
night, when hens consume less or do not consume food at all, and calcium needs
were not being met sufficiently. On of the main factors influencing the quality of
the eggshell is particle size of calcium and this problem has been present in the last
30 years, and is the subject of numerous studies. In order to overcome it,
replacement of a part of chalk in the mixture is recommended, with marble of
larger particles (Guinot and Nys 1991; Roberts and Nolan, 1997; Pavlovski et al.,
2000; Pavlovski et al., 2006; Amer et al., 2007; Pavlovski et al., 2008). Larger
marble particles slowly dissolve is in the digestive tract and calcium becomes
available during the evening and night periods. In contrast, finely ground limestone
dissolves rapidly and is used in few hours.
Phosphorus. Numerous studies show that in the rearing period, different
levels (0.2 to 0.3%) do not affect the quality of the first egg or bone mineralization.
Shell quality decreases with increased level of accessible phosphorus in the diet. In
the warm period of the year, the need for phosphorus in layers increases and if it is
below 0.25% mortality of hens and shell breakage increase. Recommended
quantity is 350 mg/day/hen to maintain skeleton of hens. The low level of
phosphorus in feed reduces the need for calcium leading to bone problems and
poor quality of eggshell. This can be partially improved by adding large particles of
marble (Nys 1995).
397

398
Z. Pavlovski et al.
Magnesium. Magnesium deficiency (less than 0.21%) influence number of
laid eggs and the shell quality. Food for laying hens should contain 0.16%, which
is four times more than the needs of laying hens (0.4%) (Vogt et al., 1984). In fact,
there is no set amount of magnesium, which is added to the diet because plant
nutrients such as bran, sunflower, rapeseed, contain enough Mg and therefore there
is no information whether, by adding of these to food, the quality of the eggshell
can be improved or worsen, except levels greater than 0.77% (Enginering, Arttech
and Leeson, 1983). In any case, the excess Mg in the diet (greater than 0.6%)
affects the increased consumption of water, which can lead to an increase in the
number of dirty eggs.
Microelements. The main trace elements to form eggshell are manganese
and copper. The lack of Cu in the diet affects negatively the biochemical and
mechanical properties of the eggshell membrane and thus leads to deformation of
egg shape (Chowdury, 1990). Mn deficit in the diet for laying hens (below 7mg)
causes thinner shell, partly due to the deterioration of see-through spots due to
worsening of the ultra shell structure in the mammalian layer and reduction of the
concentration of the polysaccharides which are precursors of matrix proteins. In the
literature there are no data that the Zn deficit in food affects the formation of the
shell, but it is known that the amount of Zn below 10mg/kg reduces the number of
fresh laid eggs. The Zn deficit in the same amount decreases the weight of hens,
egg production, the number of newly hatched chicks, hens plumage forming, but
there is no negative impact on the quality of the eggshell.
Food for laying hens containing 30mg/kg Zn, 20mg/kg Mn and 6mg/kg Cu
is not sufficient to meet the needs of hens, therefore other sources mentioned of
microelements must be provided (Zamani et al., 2005), including Fe and Co. The
deficit of these elements affects the reduction of shell mass. Abdllah et al. (1994)
consider this to be due to Mn deficiency. The amount of Mn 70-100mg/kg provides
good quality eggshell strength and thickness (Faria et al., 1999).
The content of Cu in the amount of 70-140 mg/kg reduces the eggshell
thickness. Attempts to improve the quality of the shell by adding boron (100 mg /
kg), vanadium (20 mg/kg) or fluoride (6-20 mg/kg) did not provide convincing
results. The metals such as nickel (100-500mg/kg) , chromium (500-2000mg/kg) or
lead (20-100mg/kg) reduce the mass of eggshell (Meluzzi et al., 1996).
Selenium supplementation to the diets of hens up to 0.8 mg/kg in order to
produce functional foods has no negative effect on the quality of the eggshell
(Pavlović et al., 2010).
It can be said that of all the trace elements, Mn has the leading role in
improving the strength of the shell. If it is added to hen feed in the amount of 60-
60-10 mg / kg, Mn, Zn and Cu the breaking force is increased, but not deformation
and the eggshell mass (Mabe et al., 2003).
Electrolytes. The Na (less than 0.1%) and chloride deficit (less than 0.11%)
in the diet for hens adversely affect egg production and shell quality, and on the

Shell quality – everlasting problem in ...
other hand any excess chloride in food (0.75 to 0.8%) has a detrimental effect on
the quality of the shell (Gezen et al., 2005). The presence of NaCl in water supply
for hens (2 g/l) affects the reduction in eggshell quality (Chen and Balnave, 2001).
In regard to the new system of housing of hens in accordance with EU regulations,
hens are able to get directly in touch with the waste which increases the number of
dirty eggs. In this case, if the food contains 1.6 to 5.5 g / kg of Na and 2.3 to 7.5 g
/ kg K, it results in increased water consumption and large amounts of waste and
as a consequence more dirty eggs.
Other nutrients. It is known that aluminosilicates, vitamins C and E
improve eggshell quality. Vitamin D3 in the amount of 400 IU increases the
number of eggs and improves shell quality (Whitehead, 1996). Seven (2008)
reported the positive effects of added vitamin C on the mass and egg shell
thickness in heat stress conditions, contrary to the results of research Supić et al.
(1997).
Numerous studies concerne the use of zeoilite in poultry nutrition. Sodium
aluminosilicate (zeolite, 0.75 or 1.5%) improves egg specific mass, and this was
confirmed in 77% of the 35 papers presented (Nys, 2001), especially when the Ca
level in the diet was 2.75%. There is definitely a positive effect on the quality of
the shell, and yet there are still concerns about its use in poultry nutrition and a
positive effect on the quality of eggshell. Absorption of 10 to 25% aluminum and
40% silicon in the intestines of hens originating from the sodium zeolite limits the
use of this additives (Roland et al., 1993).
Acknowledgment
Research was part of the project TR 31033, financed by the Ministry of
Education, Science and Technological Development of the Republic of
Serbia.
Ljuska – aktuelni problem u živinarstvu
Z. Pavlovski, Z. Škrbić, M. Lukić, D. Vitorović, S. Lilić, V. Petričević
Rezime
Lom ljuske (6-20%) problem je od velikog značaja sa stanovišta
ekonomskih posledica i bezbednosti jaja u ishrani. U oblasti ishrane, genetike i
uslova životne sredine se čine veliki napori kako bi se poboljšao kvalitet ljuske
jajeta, ali razumevanje procesa nastanka ljuske i porekla defekata ljuske predstavlja
399

400
Z. Pavlovski et al.
ključ za razumevanje ovog problema. Cilj ovog preglednog rada je da se rezimiraju
nova saznanja i informacije o pitanjima ljuske jajet, kao i ažuriraju novi nepovoljni
prehrambeni i uslovi životne sredine.
References
ABDALLAH, A. G., R. H. HARMS, H. R. WILSON, AND O. EL-
HUSSEINI (1994): Effect of removing trace minerals from the diet of hens
laying eggs with heavy or light shell weight. Poultry Science 73:295–301.
AMERAH A.M., RAVINDRAN V., LENTLE R.G., THOMAS D.G.
(2007): Feed particle size: Implications on the digestion and performance of
poultry. World's Poultry Science Journal, 63, 439-456.
ATTECH J. O., LEESON S. (1983): Influence of increasing dietary calcium
and magnesium levels on performance, mineral metabolism, and egg
mineral content of laying hens. Poultry Science 62: 1261-1268.
BELL D. (1998): Egg shell quality: its impact on production, processing and
marketing economics. Proceedings of Alltech's Fourteenth Annual
Symposium. Biotechnology in the Feed Industry, 447-467.
CHANDRAMONI, S., JADHAO B., SINHAR.P. (1998): Effects of dietary
calcium and phosphorus concentrations on retention of these nutrients by
caged layers. British Poultry Science 39:544–548.
CHEN J., BALNAVE D. (2001): The Influence of Drinking Water
Containing Sodium Chloride on Performance and Eggshell Quality of a
Modern, Colored Layer Strain. Poultry Science 80: 91-94.
CHOWDHURY S. D. (1990): Shell membrane system in relation to
lathyrogen toxicity and copper deficiency. World’s Poultry Science Journal
46:153–169
COUTS A.J., WILSON G.C. (2007): Optimum Egg Quality: A Practical
Approach, pp 64.
DAGHIR N.J. (2004): Nutritonal strategies to reduce heat stress in laying
hens. XXII World's Congress, Istanbul, Turkey, Book of abstracts, 289 (CD
fulltext).
FARIA D. E., JUNQUEIRAO.M., SAKOMURA N.K., SANTANA A.E.
(1999): Effect of different levels of manganese and phosphorus on the
performance and eggshell quality of laying hens. Revista Sociedade
Brasileria Zootecnia 28:105–112.

Shell quality – everlasting problem in ...
GEZEN S.S., EREN M., DENIZ G. (2005): The effect of different dietary
electrolyte balances on eggshell quality in laying hens. Revue Medecine
Veterinarie 156, 10, 491-497.
GROBAS S., MENDEZ J., DE BLAS C., MATEOS G.G. (1999):Laying
Hen Productivity as Affected by Energy, Supplemental Fat, and Linoleic
Acid Concentration of the Diet. Poultry Science 78:1542–1551.
GUINOTE F., NYS Y. (1991): Effects of particle size and origin of calcium
sources on eggshell quality and bone mineralization in egg laying hens.
Poultry Science 70, 583-592.
HAMILTON R.M.G., CIPERA J.D. (1981): Effects of Dietary Calcium Levels
During the Brooding, Rearing, and Early Laying Period on Feed Intake, Egg
Production, and Shell Quality of White Leghorn Hens. Poultry Science 60: 349-
357.
HAMILTON M. (1982): Methods and factors that affect the measurement of egg
shell quality. Poultry Science 61: 2022-2039.
HARMS D., DOUGLASR., SLOAN R. (1996): Midnight feeding of commercial
laying hens can improve eggshell quality. Journal of Applied Poultry Research 5: 1-5.
HARMS, R. H., RUSSELL G.B., SLOAN D.R. (2000): Performance of four
strains of commercial layers with major changes in dietary energy. Journal
of Applied Poultry Research 9:535–541.
KESHAVARZ K. (1998a): Further Investigations on the Effect of Dietary
Manipulation of Protein, Phosphorus, and Calcium for Reducing Their
Daily Requirement for Laying Hens 1. Poultry Science 77:1333–1346.
KESHAVARZ K. (1998b): Investigation on the possibility of reducing
protein, phosphorus, and calcium requirements of laying hens by
manipulation of time of access to these nutrients. Poultry Science 77:1320–
1332
LAVELIN I., MEIRI N., PINES M. (2000): New insight in eggshell
formation. Poultry Science 79: 1014-1017.
MABE I., RAPP C., BAIN MM., NYS Y. (2003): Supplementation of a
corn-soybean meal diet with manganese, copper, and zinc from organic or
inorganic sources improves eggshell quality in aged laying hens. Poultry
Science 8 2:1903-1913.
MELUZZI A., SIMONCINI F., SIRRI F., VANDI L., GIORDANI G.
(1996): Feeding hens diets supplemented with heavy metals (chromium,
nickel and lead). Archiv Geflügelkunde 60(3): 119-125.
NYS Y. (1995): Influence of nutritional factors on eggshell quality at high
environmental temperature. Proceedings VI European Symposium on the
Quality of Eggs and Egg Products, Zaragoza, Spain, 209-220.
401

402
Z. Pavlovski et al.
NYS Y., GAUTRON Ј., MC KEE M.D., GARCIA-RUIZ Ј.М., HINCKE
М.Т. (2000): Biochemical and functional characterization of eggshell matrix
proteins in hens. XXI World's Poultry Congress, Montreal, Canada (CD
Proceedings).
NYS, Y. (2001) Composition and nutritional value of the hen’s egg. Proceedings
9 th European Congress on the Quality of Egg and Egg Products, Kusadasi, Turkey,
325-341.
PAVLOVIĆ Z., MILETIĆ I., JOKIĆ Ž., PAVLOVSKI Z., ŠKRBIĆ Z.,
ŠOBAJIĆ S. (2010): The Effect of Level and Source of Dietary Selenium
Supplementation on Eggshell Quality. Biological Trace Element Research,
133 (2): 197-202
PAVLOVSKI Z., LUKIĆ M., ŠKRBIĆ Z., VITOROVIĆ D., CMILJANIĆ
R. (2006): Effects of additional Ca feeding on eggshell strenght in older
hens. XII European Poultry Conference, Verona, Italy. World’s Poultry
Science Journal. Book of abstracts, vol.62, 175 (CD Proceedings).
PAVLOVSKI Z., VITOROVIĆ D. (1996): Direktan metod za odredjivanje
čvrstoće ljuske jaja. Nauka u živinarstvu, 3-4, 171-175.
PAVLOVSKI Z., VITOROVIĆ D., ŠKRBIĆ Z., VRAČAR S. (2000):
Influence of limestone particle size in diets for hens and oviposition time on
eggshell quality. Acta Veterinaria, Vol. 50, No. 1, 37-42.
PAVLOVSKI Z., LUKIĆ M., ŠKRBIĆ Z., BLAGOJEVIĆ M. (2008):
Efficiency Of The Use Of Large Size Marble Particles In Conditions Of
Three Week Calcium Deficit In Young Layer Hens. 1 st Mediterranean
Summit of WPSA, Porto Carras, Greece. Book of Proceedings, 852-857.
PAVLOVSKI Z., ŠKRBIĆ Z., LUKIĆ M. (2012): Autochthonous Breed of
Chicken in Serbia: Research or Development. Proceedings of XV
International Feed Technology Symposium, Novi Sad, Republic of Serbia,
October 3-5, in press.
ROBERTS J. R., NOLAN J.V. (1997): Egg and egg shell quality in five
strains of laying hen and the effect of calcium source and hen age.
Proceedings of the VII European Symposium on the Quality of Eggs and
Egg Products, Poznan, 38-44.
ROLAND D. A., SR., BRYANT M.M., ROLAND D.A., JR., SELF J.
(1993): Econometric Nutrition IV: Maximizing profits in second cycle
commercial Leghorns (phase 1) by optimizing feeding methods, total sulfur
amino acid (TSAA) intake and environmental temperature (ET). Poultry
Science 72 (Suppl. 1): 53.
ROLAND D.A. (1988): Eggshell problems: Estimates of incidence and
economic impact. Poultry Science 67, 1801-1803.

Shell quality – everlasting problem in ...
ROLAND D.A. (2000): Nutrition and feeding for optimum egg shell
quality. XXI World’s Poultry Congress, Montreal, Canada (CD
Proceedings).
SAFAA H.M., SERRANO M.P., VALENCIA D.G., FRIKHA M.,
JIMÉNEZ-MORENO E., MATEOS G.G. (2008): Productive Performance
and Egg Quality of Brown Egg-Laying Hens in the Late Phase of
Production as Influenced by Level and Source of Calcium in the Diet.
Poultry Science 87:2043-2051.
SCOTT T. A., KAMPEN R., SILVERSIDES F.G. (1999): The effect of
phosphorus, phytase enzyme, and calcium on the performance of layers fed
corn-based diets. Poultry Science 78:1742–1749.
SEVEN P.T. (2008): The Effects of Dietary Turkish Propolis and Vitamin C
on Performance, Digestibility, Egg Production and Egg Quality in Laying
Hens under Different Environmental Temperatures. Asian-Australian
Journal of Animal Science 21: 1164-1170.
SUPIĆ B., CMILJANIĆ R., SAVIĆ S., MILOŠEVIĆ N., KOČIŠ I.,
JAKOBČIĆ Z. (1997): Uticaj vitamina C na proizvodnju i kvalitet ljuske
jaja za konzum. I Jugoslovenski medjunarodni kongres o stočarstvu.
Biotehnologija u stočarstvu 3-4, 177-186.
ŠKRBIĆ Z., PAVLOVSKI Z., HOPIĆ S., VRAČAR S., LUKIĆ M. (1998.):
Uticaj vremena ovipozicije i sprata baterije na kvalitet ljuske jaja. Nauka u
živinarstvu, 1-2, 207-210.
TUMOVA E., ZITA L., HUBENY M., SKRIVAN M., LEDVINKA Z.
(2007): The effect of oviposition time and genotype on egg quality
characteristics in egg type hens. Czech Journal of Animal Science, 52, 26–30.
VITOROVIĆ D., PAVLOVSKI Z., NIKOLOVSKI J., DJURDJEVIĆ Z.,
TODOROVIĆ M. (1995): Kvalitet ljuske i dalje aktuelan problem
savremenog živinarstva. IV Medjunarodni simpozijum "Novi pravci razvoja
stočarstva", Beograd, Biotehnologija u stočarstvu 3-6, 301-306.
VOGT X., DEWAR W.A., SAUVEUR B., SIMONS P.C.M. (1984):
Mineral requirements and recommendations for adult birds. World's Poultry
Science Journal 40, 183.
WASHBOURN K. (1982): Incidence, cause and prevention of egg shell breakage
in commercial production. Poulttry Science 61: 2005-2012.
WHITEHEAD C.C. (1996): Nutrition and bone disorders in poultry.
Proceedings of XX World’s Poultry Congress, New Delhi, Vol. II, 161-171.
403

404
Z. Pavlovski et al.
ZAMANI A., RAHMANI H.R., POURREZA H.R. (2005): Eggshell quality
is improved by excessive dietary zinc and manganese. Proceedings of 15 th
European Symposium on Poultry Nutrition, Balatonfured, Hungary, 520-522.
ZHANG L.C., NING Z.H., XU G.Y., HOU Z.C., YANG N. (2005):
Heritabilities and genetic and phenotypic correlations of egg quality traits in
brown-egg dwarf layers. Poultry Science 84: 1209-1213.
Received 14 April 2012; accepted for publication 21 August 2012

Biotechnology in Animal Husbandry 28 (3), p 405-414 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.034
DOI: 10.2298/BAH1203405M
EFFECT OF FEATHERING ALLELES (K/K + ) ON LAYING
PERFORMANCE, HATCHABILITY PARAMETERS AND
SOME BODY MEASUREMENTS IN TWO LINES OF
WHITE PLYMOUTH ROCK HENS
N. Mincheva, M. Lalev, M. Oblakova, P. Hristakieva, I. Ivanova
Agricultural Institute - Stara Zagora, Bulgaria
Corresponding author:nmincheva@abv.bg
Original scientific paper
Abstract: The study aimed to investigate the effects of sex – linked
feathering alleles on laying performance, hatchability parameters and body
measurements in hens from two White Plymouth Rock lines (line L and line K)
used as maternal lines in broiler production. Four groups of 18-week-old hens were
formed, two of each line, with genotype K/W (slow feathering) and k + /W (rapid
feathering) respectively. Groups of line L included 72 hens divided into 6 boxes
with 1 rooster per 12 hens, whereas line K groups comprised 96 hens of each
genotype, housed in 8 boxes with one rooster per 12 hens, totally 192 birds.The K
locus alleles had no significant effect on egg production traits (p>0.05). The
presence of slow feathering allele resulted in lower fertility and hatchability of set
eggs in both studied lines (p

406
N. Mincheva et al.
sexing is more efficient and less stressful (Harris et al., 1984,; Wilson et al., 2007).
For this purpose, roosters with k + /k + genotype are mated to K/W hens, resulting in
slow feathering of one-day-old male chicks and rapid feathering in female one-dayold
chicks. The autosexing allows a rapid identification of the sex of chicks without
any special training or sexoscopes, with minimum inaccuracy.
The utilisation of the dominant K allele (slow feathering) in the maternal
line for production of autosexing chicks distinguished by feathering rate could, in
the belief of some researchers, be linked to an unfavourable effect on sexual
maturity (Verma and Singh, 1983; Harris et al., 1984; O’Sullivan et al., 1991;
Ning et al., 2005; Abd El-Rahman, 2006), egg production (Lowe and Garwood,
1981; Saleh et al., 1987; Havenstein et al., 1989; Abd El-Rahman. 2006; Wilson et
al., 2007; Durmus et. al, 2010), livability (Bacon et al., 1988; Alsobayel and Al –
Muhlem, 1997), feed conversion (Белоречков & Алиуи, 1986) and reproduction
(O’Sullivan et al., 1991), although others did not show any differences between
slow- and rapid feathering hens in these parameters (Dunnington and Siegel, 1986;
Ning and Chen; 1988; Prijono and Smith, 1994; Холодков, 2004; Younis and
Galal, 2006; Ledvinka et al., 2011).
In the view of Chambers et al. (1993) the contradictory results suggest an
interaction of the K allele with other genes from the genome. For instance, Bacon
et al. (1988) established a close relationship between the K slow feathering gene
and ev 21 gene in a White Leghorn line and this way explained the lower egg
productivity and livability in the offspring of birds with the slow feathering
genotype. On the other side, Dunnington and Siegel (1986) outline as a possible
cause for contradictory results the effect of K gene on some biological factors such
as heat insulation properties of vane feathers, and the genetic origin of studied
populations.
Our observations have demonstrated the presence of a slow feathering K
gene in two White Plymouth Rock lines. One of the main prerequisites of using
marker K locus genes is the lack of negative impact on production traits of hybrids,
and therefore, the knowledge on effects of K/k + alleles on productivity is essential
for breeding programmes.
Taking into consideration the contradictory results, we aimed to establish
the effect of sex-related alleles for feathering rate on some production traits,
morphological and incubation traits of eggs and exterior traits in hens from these
two lines.
Materials and methods
The experiment was performed in the selection base of the Poultry
Breeding Unit at the Institute of Agriculture – Stara Zagora in 2009–2010.

Effect of feathering alleles (k/K + ) on laying performance ...
Two original White Plymouth Rock lines of chickens were used as maternal forms
for broiler production – line L and line K.
In the progeny of these lines, the sex of day-old chicks was determined
with sexoscope, the feathering rate was evaluated according to the wing feathering
speed, and the chicks were wing - banded. At the age of 10 days their tail feathers
were checked. Chicks without tail feathers were slow feathering. For line L, the
frequency of slow and rapid feathering genotypes in female chicks was 79.70% and
20.30%, respectively. In female chicks from line K, the rapid feathering genotype
was encountered at a frequency of 69.40%, whereas the slow feathering genotype –
at 30.60%. Chicks from each line and genotype were housed uniformly, on a deep
wood shavings litter according to the technology requirements implemented in the
selection base until the age of 18 weeks. At that time, four groups were formed –
two of each line with genotypes K/W (slow feathering) and k + /W (rapid
feathering). Groups of line L included 72 hens divided into 6 boxes with 1 rooster
per 12 hens, whereas line K groups comprised 96 hens of each genotype, housed in
8 boxes with one rooster per 12 hens, totally 192 birds.
The hens were housed until 48 weeks of age on deep litter with uniform
conditions with regard to density, feeding and drinking widths. Restricted feeding
was practiced with weekly rations according to the age and egg production. The
compound feed contained: metabolizable energy 1810.005 kcal/kg, crude protein –
16.012%, crude fat – 6.836%, crude fibers – 5.889%, lysine 0.75%, methionine
0.38%, calcium 3.2%, phosphorus 0.81%
During the experiment, the following traits were evaluated:
• Age of sexual maturity (days) – age when 50% of egg production
was attained in the different groups;
• Egg production – daily from egg lay beginning to 48 weeks of age.
On the basis of these data, the hen-day and hen-housed egg production
percentages were established;
• Average egg weight – by weighing eggs laid by each group for the
day at 2-week interval between 32 and 48 weeks of age
• Morphological traits of eggs – 10% of the daily egg yield during 3
consecutive days at 40 weeks of age was analysed. The individual egg
weight, shape index, albumen index, yolk index, Hough units, yolk
colour by the La Roche scale and eggshell thickness were determined.
• Incubation traits of eggs – they were investigated at 48 weeks of age
by collection of eggs over 7 days. The fertility was evaluated as the
relative share of fertilized to eggs set in the incubators; the hatchability
of eggs set (HS) – the relative share of hatched chickens from eggs set;
the hatchability of fertile eggs (HF) – the relative share of hatched
chickens from fertilised eggs, embryonic death rate – for the
407

408
N. Mincheva et al.
incubation intervals 0–6 days, 7–18 days and 19–21 days – as relative
proportions of dead embryos from fertilised eggs.
• Live body weight and exterior traits at 36 weeks of age – the live
body weight was determined by weighing on a balance with precision
of 5 g. The following body parts were measured with a centimetre
tape: body length (between the last cervical vertebra and the tail root),
breast circumference (between the last cervical vertebra behind and
under the wings to the anterior end of the keel bone ridge); keel bone
length (between the anterior and posterior ends of keel bone ridge),
thigh, drumstick and metatarsus lengths (between the end points of
respective bones); the chest depth between the last cervical vertebra
and the anterior keel bone border) – with compasses.
Data were statistically processed by lines and by genotypes by means of
ANOVA/MANOVA and LSD post hoc test with Statistica 8 software (StatSoft,
2009). Percentage data were arcsine transformed prior to be analysed.
Results and discussion
The analysis of results in table 1 showed that the age of sexual maturity in
hens from line L did not differ significantly between both genotypes as also
reported by Holodkov (2004), Ledvinka et al. (2011). Тhis tendency was not valid
for line K, where the beginning of egg lay in the group with rapid feathering
genotype k + /W occurred 6 days earlier (p

Effect of feathering alleles (k/K + ) on laying performance ...
The hen-housed and hen-day egg production, the feed expenditure per one
egg produced and the livability during the period of lay were not influenced by the
feathering rate genotype. With this regard, our results confirmed the conclusions of
Prijono and Smith (1994), Holodkov (2004) and Younis and Galal (2006).
Table 2 presents the data from ANOVA statistical analyses about the effect
of K locus genotype on morphological traits of eggs. The feathering genotype had a
significant impact on the shape index (p

410
N. Mincheva et al.
pronounced effect of K locus alleles on fertility, early embryonic death and
hatchability of fertile eggs, but unlike our results, the fertility, early and late
embryonic death rates were higher in slow feathering birds, whereas the
hatchability of fertile eggs – in rapid feathering birds. The hatchability of incubated
eggs was similar. On the other hand, Alsobayel and Al – Muhlem (1997) did not
report any significant differences with regard to fertility rate, hatchability and
embryonic death rate between slow and rapid feathering Baladi chickens.
Table 3. Hatchability traits of hens as affected by feathering genotype
Traits
Line
K L
Feathering genotype
k + /W K/W k + /W K/W
Significance
P - value
Fertility (%) 92,88a 86,73b 92,19a 85,84b 0,022
Early embryonic
0,42 0,00 0,68 1,40 0,631
mortality (%)
Middle period
6,29 3,75 5,73 6,76 0,701
embryonic mortality (%)
Late embryonic
3,61 8,28 5,52 4,56 0,302
mortality (%)
Hatchability
92,65 92,98 94,66 89,94 0,364
/fertile eggs/, %
Hatchability
87,50a 80,77b 88,79a 81,01b 0,044
/set eggs/, %
a–b statistically significant differences (p

Effect of feathering alleles (k/K + ) on laying performance ...
Table 4. Body weight and body measurements of hens at 36 weeks of age
Traits
Line
K L
Feathering genotype
k + /W K/W k + /W K/W
411
Significance
P - value
Body weight (g) 3874,7a 3546,20b 3327,90c 3377,14c 0,000
Body length (cm) 21,30a 21,17a 19,90b 20,70b 0,346
Breast circumference (cm) 39,83 38,30 39,70 39,07 0,080
Breast depth (cm) 12,67a 12,60a 12,90a 11,93b 0,081
Keel length (cm) 14,03a 14,30a 13,37c 14,23a,b 0,050
Thigh length (cm) 14,03a 14,23a 13,20b 14,30a 0,003
Drumstick length (cm) 15,90a 16,20a 14,40b 14,83b 0,055
Metatarsus length (cm) 8,43d 9,03c 9,60b 10,13a 0,000
a–d statistically significant differences (p0.05).
The presence of slow feathering allele resulted in lower fertility and hatchability of
incubated eggs in both studied lines (p

412
N. Mincheva et al.
proizvodnji brojlera. Četiri grupe kokoši nosilja uzrasta 18 nedelja, dve grupe po
liniji, genotipova K/W (sporo operjavanje) i k + /W (brzo operjavanje), respektivno.
Grupe linije L su sadržavale 72 kokoši podeljene u 6 bokseva sa 1 petlom na 12
kokoši, dok su grupe linije K imale 96 kokoši svakog genotipa, smeštene u 8
boksova sa jednim petlom na 12 kokoši, ukupno 192 grla.
K locus alela nije imao značajan uticaj na osobine proizviodnje jaja (p>
0,05). Prisustvo alela za sporo operjavanje je rezultiralo u slabijoj plodnosti i
procentu izleženih jaja u obe ispitivane linije (p

Effect of feathering alleles (k/K + ) on laying performance ...
Production and Hatchability Parameters. Asian Journal of Animal and Veterinary
Advances 5 (1): 66-71.
HARRIS, D. L., V. A. GARWOOD, P. C. LOWE, P. Y. HESTER, L. B.
CRITTENDEN, A. M. FADLY. (1984): Influence of sex-linked feathering
phenotypes of parents and progeny upon lymphoid leukosis virus infection status
and egg production. Poult. Sci. 63:401–413.
HARRIS, D. L., V. A. GARWOOD, P. C. LOWE, P. Y. HESTER, L.
B.CRITTENDEN, FADLY, A. M. (1984): Influence of sex-linked feathering
phenotypes of parents and progeny upon lymphoid leukosis virus infection status
and egg production. Poult. Sci. 63:401–413.
HAVENSTEIN, G.B., V. D. TOELLE, R. H. TOWNER, A. EMSLEY. (1989):
Effects of genetic strain, slow verus rapid - feathering maternal genotype and cage
density on the performance of Single Comb White Leghorns. Poultry Sci.68 : 596-
607.
HOLODKOV, V. (2004): Productivity of meat type hens carrying of the K and k
genes Scientific and practical experience in poultry breeding (VNITP) 1. Sergiev
Posad, pp. 4-5
HUTT, F.B. (1949): Genetics of fowl. McGraw, Hill Book Company, Inc. New
York
LEDVINKA, Z, L. ZITA, M. HUBENY, E. TUMOVA, M. TYLLER, P.
DOBROVOLNY, M. HRUSCA. (2011): Effect of genotype, age of hens and K/k
allele on egg shell quality. Czech. J. Anim. Sci., 56 (5): 242-249
LOWE, P. C. and V. A. GARWOOD. (1981): Independent effects of K and k +
alleles and maternal origin on mortality and performance of crossbred
chickens.Poult. Sci. 60: 1123 – 1126
NING, G. J. and S. Q.CHEN. (1988): The influence of genotype on the
performance of layers. Chinese Journal of Animal Science No. 3 pp. 3-5
NING, Z. H., W. ZONG, P. DANFANG, H. ZHUOCHENG. (2005): Effect of
avian leukosis and sex-linked feather gene on performance of White Leghorn hens.
Chinese Journal of Animal Science Vol. 41 No. 10 pp. 23-25
O’SULLIVAN, N. P., E. A. DUNNINGTON, E. J. SMITH, W. B. GROSS, P. B.
SIEGEL. (1991): Performance of early and late feathering broiler breeder females
with different feeding regimens. Br. Poult. Sci. 32:981–995.
PRIJONO, S. N. and W. K., SMITH. (1994): The effect of divergent selection for
the performance of broiler chickens. Sustainable animal production and the
environment. Proceedings of the 7th AAAP Animal Science Congress, Bali,
Indonesia, 11-16 July, 1994. Volume 2: pp 197-198
SALEH, K., FARGHALY, M. ,ALI M. M.. (1987): The effect of feathering rate and
housing system on some economic traits of layers. Archiv. Geflugelk. 51: 153 – 157.
SEREBROVSKY, A. S. (1922): Crossing-over involving three sexlinked genes in
chickens. Am. Nat. 56:571–572.
413

414
N. Mincheva et al.
VERMA, S. K. and P. SINGH. (1983): Effect of slow and rapid – feathering genes
on economic traits in WL x RIR crosses. Indian J. Anim. Sci. 53: 105 – 106
WILSON, H.R., J.P. JACOP, F. B. MATHER. (2007): Method of sexing day – old
chicks. University of Florida, Cooperative Extension Service, USA
YOUNIS, H. H. and A.GALAL. (2006): Impact of dwarf (dw), rapid feathering
(k + ) and naked neck (Na) genes on growth and egg production parameters of laying
hen chickens. Egyptian Poultry Science Journal, Vol. 26 No. 1 pp. 17-38
Received 14 August 2012; accepted for publication 21 August 2012

Biotechnology in Animal Husbandry 28 (3), p 415-424 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.085.8
DOI: 10.2298/BAH1203415T
THE EFFECT OF PHASE NUTRITION DURING
STARTER PERIOD ON PRODUCTION PERFORMANCES
AND NITROGEN CONTENT IN FECES OF BROILERS
OF DIFFERENT GENOTYPES
N. Tolimir 1 , L. Perić 2 , N. Milošević 2 , M. Đukić – Stojčić 2 , R. Jovanović 1 ,
V. Bogdanović 3
1 ,
Institute of Science Application in Agriculture Bulevar Despota Stefana 68b, 11000, Belgrade,
Republic of Serbia
2
Faculty of agriculture, University of Novi Sad, Trg Dositeja Obradovića 8, 21000, Novi Sad,
Republic of Serbia
3
Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade, Republic of Serbia
Corresponding author: e-mail: ntolimir@ipn.co.rs
Original scientific paper
Abstract: Objective of the research was to investigate the effect of phase
nutrition, i.e. different mixtures used in broiler nutrition with phase reduction of
protein cotnent during starter period on production peformances of fattening
chickens of diferent genotypes and on nitrogen content in faeces. Differences
between groups (treatments) were in the type of mixtures use din starter period, and
according to the following program: T1 (control group) – nutrition consisted of
mixtures with 23% of proteins in duration from day 1 to 21; T2 – from day 1 to 14,
mixture containing 23% of protein was used, and from day 14 to 21, mixture with
20% of proteins; T3 – from day 1 to 7, mixture was used containing 23% of protein
and from day 7 to 21, mixture with 20% of proteins; T4 – nutrition with mixture
containing 20% of proteins in duration from day 1 to 21. During the trial period
(from day 1 to 21) production parameters were followed. Nitrogen content was
determined in a collective faeces sample. Results showed that chickens of different
genotypes expressed different sensitivity to protein restriction. Comparing the trial
groups with the control in the third week, in chickens of Ross 308 provenience
significantly lower body weight was registered only in chickens of T4 group with
the highest protein reduction. In Cobb 500 chickens, significantly lower body
weight was determined in T3 and T4 trial groups. In case of both hybrids, in T2
group, slightly lower body weight was established, but without statistically
significant difference compared to the control. Feed conversion in both genotypes
was the highest in T3 and T4 treatments. The program of phase nutrition resulted in
decrease of the nitrogen content in faeces. Based on obtained data it can be
concluded that applied nutrition treatments with drastic protein reduction during
starter period in both genotypes had negative effect on production performances.

416
N. Tolimir al.
Therefore, the composition and dynamics of mixture changes must be taken into
consideration when formulating the program of phase nutrition, in order to achieve
both goals – decrease of protein content in feed with simultaneous diminished
nitrogen excretion and achieving good results.
Key words: broiler, genotype, phase nutrition, production traits, nitrogen
Introduction
In regard to broiler fattening periods, the early period is given special attention,
and nutrition during the starter period is considered critical for realization of
optimum production performances, therefore is the topic of research by numerous
authors (Baker and Han, 1994; Gomes et al., 2006). Studies show that the protein
and amino acid requirements of broilers change with their age, also that the use of
one diet during longer period of time would result either in nutrient suficit or
deficit during most of the growth period. In this regard, Belyavin, (1999) suggests
that during the growing period more different diets are used, i.e. that nutrition of
broilers is based on programs of multiphase nutrition. Studies show that mixtures
with reduced protein content have no effect on body weight and feed intake of
broilers, i.e. their impact on economical efficiency of broiler fattening is significant
(Warren and Emmert, 2000; Saleh et al., 1996). Numerous programs of phase
nutrition have been developed in the last decade which are based on different
frequencies of changes of certain mixtures, i.e. starter, grower and finisher in
broiler nutrition. Main objective of phase nutrition programs is to determine the
optimum time proportion in nutrition using basic mixtures, in regard to production
performances and economical efficiency of production, but often also the
ecological aspect of these programs is considered (Roush et al., 2004; Saleh et al.,
2004). Also, it can be observed that programs include certain combinations of
minimum three to twenty mixtures of different nutritional composition (Buteri et
al., 2009; Tolimir et al., 2010), with aim to determine the optimal program.
Economical impact of substitution starter with grower mixture in early period is
based on the difference in cost of starter and finisher diets. In addition to the
economical impact of the phase nutrition program, the studies include also the
ecological aspect, considering that it was established that nitrogen excretion is
directly correlated to protein content in mixtures used in nutrition (Bregendahl et
al., 2002). However, study results (Ferguson et al., 1997) indicate that drastic
reduction of crude proteins during the first three weeks can significantly increase
the need for feed in broilers, therefore it is not recommended, although reduction of
crude proteins resulted in decrease of nitrogen emission. The same author
(Ferguson et al., 1998) points out that feed manipulation can have impact on

The effect of phase nutrition during ...
reduction of nitrogen in manure, but at the same time production performances are
maintained at the acceptable level.
Objective of the present paper is to investigate the effect of the nutrition
program which includes several phases, i.e. the effect of different mixtures used in
broiler nutrition, where protein content is reeduced in phases – in the first fattening
phase, on production traits of fattening chickens of different genotypes and
nitrogen content in faeces.
Material and Methods
Trial was carried out on Experimental farm of the Faculty of Agriculture, Novi
Sad in Temerin. The study included 1200 individually marked chickens of Ross
308 and Cobb 500 proveniences. Chickens were housed in 16 boxes, 75 chickens
in each box. Within each genotype, four groups – treatments were formed, with
two repetitions for each treatment. For every treatment 150 chickens per treatment
were studied.
Standard fattening technology was applied, in duration of 42 days. Chicken
nutrition was ad libitum. Difference between groups (treatments) was in the type of
mixture used in starter period, and according to the following program of phase
nutrition: T1 (control group) – nutrition consisted of mixtures with 23% of proteins
in duration from day 1 to 21; T2 – from day 1 to 14, mixture containing 23% of
protein was used, and from day 14 to 21, mixture with 20% of proteins; T3 – from
day 1 to 7, mixture was used containing 23% of protein and from day 7 to 21,
mixture with 20% of proteins; T4 – nutrition with mixture containing 20% of
proteins in duration from day 1 to 21.
Control measuring of all individually marked chickens was done using
technical scale on the first day and in weekly intervals from week 1 to 3. Feed
conversion was calculated based on data on feed intake and gain of chickens at the
level of every chicken box, and all dead chickens were also weighed and included
in the calculation.
Chemical analysis was carried out on the collective faeces sample, as well as
determination of nitrogen content. Collective faeces sample was formed by
collecting faeces from trial boxes for studied treatments. Faeces were collected
using a wooden box with wire basis on which a nylon layer was placed, and where
chickens were kept for certain period of time necessary for taking of samples.
Nitrogen content in faeces was analyzed in the Laboratory for quality of animal
feed of the Faculty of Agriculture in Novi Sad, according to the method 7 of the
Rulebook on methods for taking of samples and methods of physical, chemical and
microbiological analysis of animal feed (1987).
The program STATISTIKA, ANOVA MANOVA and LSD post-hoc test were
used in data processing.
417

418
Results and Discussion
N. Tolimir al.
Based on obtained data, it was established that applied program of phase
nutrition during starter period, in both investigated proveniences, resulted in
expression of impact on production performances, i.e. body weight and feed
conversion.
In Table 1, the weekly evaluation of the significance of differences of major
factors on body weight of Cobb 500 and Ross 308 chickens is presented. Hybrid
was determined to be significant only in the first week, which can be associated
with the research by Petričevič et al. (2011), whereas the feed as variability factor
had statistically significant effect on body weight in all trial weeks. Interaction
between factors hybrid and feed was not recorded in the third week.
By observing the data obtained in different nutrition treatments, it is apparent
that hybrids reacted differently on applied nutrition programs. In case of
provenience Cobb 500 significant difference between the control – T1 compared to
chickens of group T3 which consumed starter mixture only in the first 7 days and
T4 group which did not consume starter diet during first three weeks, was
established. In Ross 308 chickens, statistically significant difference was
determined only between T1 and T4 groups, which can indicate that chickens of
this provenience were more tolerant to shortening of the nutrition with starter
mixture.
Table 1. Evaluation of significane of differences in body weight, per weeks
Week
Variation
measure
Average body weight (g)
Cobb 500 Ross 308 Factor
T1 T2 T3 T4 T1 T2 T3 T4
1. x 144
Sd
a
147
18,47
a
143
17,18
a
118
19,76
b
159
11,25
a
161
17,92
a
156
18,84
a
126
19,29
b
** ** *
16,04
2. x 333
Sd
a
333
58,82
a
315
42,07
b
262
40,98
c
336
38,99
a
346
51,71
a
315
54,05
b
283
52,07
c
NS ** **
43,09
3. x 642
Sd
a
621
116,54
a
614
87,12
b
530
85,87
c
642
92,08
a
638
93,28
a
623
104,57
a
554
100,71
b
NS ** NS
94,54
a-b Values within the row for each hybrid without same letter in superscript are statistically significantly different
(P

The effect of phase nutrition during ...
during the investigation of the impact of nutrition program was alo reported by
Mandrigal et al. (1994), Nikolova et al. (2007). However, in the paper by Saki et
al. (2010) in the investigation of three programs of nutrition during starter period in
two genotypes, no significant interactions between hybrids and nutrition programs
were established.
In case of proveniences Cobb 500 and Ross 308, the lowest, insignificant
difference in final body weight was registered by comparing T1 with T2 group,
which consumed starter mixture during the period of two weeks. This result induces
the question if chickens need starter mixtures longer than 14 days, during starter
period. Obtained results are in accordance with research by Saleha et al. (1997) who
evaluated the time proportion in nutrition using starter, grower and finisher mixtures
in fattening chickens in fattening period of 42 days. This author indicated the
significance of target body weight when planning the nutrition program in sense of
duration of certain nutrition phases and provided recommendations for duration of
use of certain mixtures depending on whether the chickens are fattened to 1kg, 2kg
or 3kg of final weight.
Obtained results can be associated with studies by Saleh et al.( 1996); Warren
and Emmert (2000); Pope and Emert (2001) who pointed out the need for reevaluation
of the traditional programs of nutrition and duration of each nutrition
phase, based on their research of the effect of phase nutrition programs and different
time proportions for starter, grower and finisher mixtures, and also taking into
consideration the genetic improvements in broilers and reduced time for realization
of body weight, as well as more expressed need for optimization of production from
economical aspect.
Table 2 presents the data on feed conversion during the first fattening phase,
for proveniences Cobb 500 and Ross 308.
During starter period (from 1 to 21 days), in chickens of both proveniences, by
comparing values obtained for feed conversion, i.e. by comparing control T1 group
to trial groups, it was observed that chickens of groups T3 and T4, with the greatest
protein reduction, realized less favourable conversion. Also, in both hybrids, feed
conversion values for T1 control group and T2 group with the mildest reduction of
protein content were similar.
Table 2. Feed conversion in chickens according to fattening phases
Period
Feed conversion
Cobb 500 Ross 308
(days) T1 T2 T3 T4 T1 T2 T3 T4
1-21 1,643 1,658 1,897 2,233 1,648 1,659 1,888 1,942
In the analysis of obtained data on feed conversion it was established that the
most favourable feed conversion was in T1 group. Obtained results are in
concordance with results obtained by Saki et al. (2010), in the research of different
419

420
N. Tolimir al.
nutrition programs. However, in the paper by Saleha et al. (1997) in phase nutrition
of broilers, no significant differences in feed conversion were detected between
NCR program and two trial programs in which chickens consumed starter mixture
to the age of 7 and 14 days.
Content of dry matter and nitrogen in faeces of Cobb 500 and Ross 308
chickens is presented according to nutrition treatments and weeks during the starter
period, i.e. from week 1 to 3 (Table 3). Also, dynamics in changing of mixtures
used in nutrition of studied genotypes depending on the nutrition program, is
presented.
By analyzing data presented in Table 3, it can be observed that content of dry
matter in chicken faeces varied depending on the feed consumed by chickens. In
both investigated proveniences, dry matter content in faeces was mainly higher
when grower mixture was consumed.
Nitrogen content in faeces, in chickens of Cobb 500 and Ross 308, was the
highest in control T1 group which consumed during all three weeks mixtures
containing 23% of crude proteins, i.e. group without protein reduction. Also, it can
be observed that the drastic reduction of protein in mixture used in group T4 in
both studied genotypes resulted in reduced nitrogen content in faeces, which is
desirable from the ecological aspect. However, it should be pointed out that this
program of nutrition had adverse effect on growth rate of chickens and feed
conversion ratio, which makes it unacceptable from the aspect of production
performances.
Table 3. Dry matter and nitrogen content in faeces
Week
1.
2.
3.
Trait
T1 starter
T2 starter
Dry matter and nitrogen content in faeces (%)
Cobb 500 Ross 308
T3 starter
T4 grower
T1 starter
T2 starter
T3 starter
T4 grower
Dry mat. 19,68 20,28 19,92 22,88 18,85 19,06 19,24 22,02
Moisture 80,32 79,72 80,08 77,12 81,15 80,94 80,76 77,98
N in DM 5,59 5,35 5,28 4,01 4,39 5,15 4,78 3,86
T1 starter
T2
starter
T3
grower
T4
grower
T1
starter
T2
starter
T3
grower
T4
grower
Dry mat. 18,68 19,95 20,53 19,78 16,67 18,26 20,09 19,46
Moisture 81,32 80,05 79,47 80,22 83,33 81,74 79,91 80,54
N in DM 5,01 4,98 4,02 3,88 4,94 5,10 4,18 3,98
T1 starter
T2
grower
T3
grower
T4
grower
T1
starter
T2
grower
T3
grower
T4
grower
Dry mat. 18,24 21,83 19,98 21,19 17,62 21,86 20,90 21,68
Moisture 81,76 78,17 80,02 78,81 82,38 78,14 79,10 78,32
N in DM 5,23 4,50 4,25 4,67 5,05 4,38 4,10 4,56
Similar results were obtained by Ferguson et al. (1997), in their study of the
effect of nutrition with different content of crude proteins on growth, concentration

The effect of phase nutrition during ...
of ammonia and manure composition. According to the results of said authors,
drastic reduction of protein content during starter period increased significantly the
need for feed, therefore it is not recommended, even though the reduction of
protein induced reduced nitrogen emission.
Obtained results can be associated with the research in which the correlation
between the protein content of mixtures used in broiler nutrition and nitrogen
excretion was confirmed (Delezie et al., 2009; Namroud et al., 2008). Linear trend
in increase of ammonia emission with the increase of crude protein content in diets
was also established by Elwingwe and Svensson (1996). This indicates the need for
further investigation of the nutrition program with comprehensive analyses in sense
of economical and ecological aspects.
Conclusion
Phase nutrition included application of nutrition program with different
frequency of changing starter, grower and finisher mixtures used in nutrition of
Cobb 500 and Ross 308 chickens, with aim to determine the optimum for
production in the economical and environmental sense.
Drastic reduction of proteins in T4 nutrition program resulted in significant
decrease of body weight and unfavourable feed conversion in both studied hybrids.
Chickens of Cobb 500 provenience were more sensitive to protein reduction. At the
same time, drastic reduction of proteins in T4 nutrition program, in both studied
genotypes, resulted in decrease of nitrogen content in faeces, which is desirable
from ecological aspect. However, negative impact of this program on production
performances makes it unacceptable. In proveniences Cobb 500 and Ross 308, the
least, insignificant difference in body weight and similar feed conversion were
established by comparing the control to T2 group, which consumed starter mixture
for the period of two weeks, indicating the need for further research of this
nutrition program.
Based on obtained results, it can be concluded that, composition and dynamics
of changes of mixtures when formulating the program of phase nutrition must be
taken into consideration in order to achieve both goals – decrease the protein
content in feed, with diminished excretion of nitrogen in faeces and realizing good
production results at the same time.
Acknowledgment:
Research was financed by the Ministry of Education, Science and Technological
Development Republic of Serbia, project TR-31033.
421

422
N. Tolimir al.
Uticaj fazne ishrane u starter periodu na proizvodne
performanse i sadržaj azota u fecesu brojlera različitih
genotipova
N. Tolimir, L. Perić, N. Milošević, M. Đukić – Stojčić, R. Jovanović, V. Bogdanović
Rezime
Cilj istraživanja je da se ispita uticaj fazne ishrane, odnosno različitih smeša za
ishranu brojlera u kojima je sadržaj proteina smanjen fazno u starter periodu na
proizvodne osobine tovnih pilića različitog genotipa i sadržaj azota u fecesu.
Razlika između grupa (tretmana) bila je u tipu smeša u starter periodu, a prema
sledećem programu: T1 (kontrolna grupa) - ishrana smešom sa 23% proteina u
trajanju od 1. do 21. dana; T2 - ishrana od 1. do 14. dana smešom sa 23% proteina i
od 14. do 21. dana sa 20% proteina; T3 - ishrana od 1. do 7. dana smešom sa 23%
proteina i od 7. do 21. dana sa 20% proteina; T4 - ishrana smešom sa 20% proteina
u trajanju od 1. do 21. dana. U oglednom periodu (od 1. do 21. dana) praćeni su
proizvodni parametri. Sadržaj azota u fecesu određen je na zbirnom uzorku.
Rezultati su pokazali da su pilići različitih genotipova ispoljili različitu
osetljivost na restrikciju proteina. Poređenjem oglednih grupa sa kontrolnom u
trećoj nedelji, kod pilića provenijence Ross 308 konstatovano je da je signifikantno
manju telesnu masu ostvarila samo T4 grupa sa najstrožijom redukcijom proteina.
Kod provenijence Cobb 500 signifikantno manja telesna masa utvrđena je za T3 i
T4 oglednu grupu. Kod oba hibrida za T2 oglednu grupu utvrđena je nešto manja
telesna masa, ali bez statistički značajne razlike u poređenju sa kontrolnom.
Konverziju hrane kod oba genotipa imala je najveće vrednosti za T4 i T3 tretman.
Programi fazne ishrane rezultirali su smanjenjem sadržaja azota u fecesu.
Na osnovu dobijenih rezultata može se zaključiti da su primenjeni tretmani
ishrane sa drastičnom restrikcijom proteina u starter periodu kod oba genotipa
imali negativan uticaj na proizvodne performanse. Zbog toga se prilikom
formulisanja programa fazne ishrane strogo mora voditi računa o sastavu i dinamici
promene smeša, a kako bi se postigla oba cilja – smanjenje sadržaja proteina u
hrani uz smanjenu ekskreciju azota i postizanje dobrih proizvodnih rezultata.
References
BAKER D.H., HAN Y. (1994): Ideal amino acid profile for chicks during the first
three weeks post hatching. Poultry Science 73: 1441-1447.

The effect of phase nutrition during ...
BELYAVIN C.G. (1999): Nutrition management of broiler programs. Animal
Nutrition. Nottingham University Press, Nottingham, Recent Adv UK. Pages 93-
105
BREGENDAHL K, SELL J.L., ZIMMERMAN D.R. (2002): Effect of low-protein
diets on growth performance and body composition of broiler chicks. Poultry
Science 81: 1156-1167.
BUTERI C.B., TAVERNARI F. DE C., LELIS G.R, ROSTAGNO H.S., ALBINO
L.F.T. (2009): Effects of different nutritional plans on broiler performance. Revista
Brasileira de Ciencia Avicola 11 no 4.
DELLEZIE E., MAERTENS L., HUYGHEBAERT G., LIPPENS M. (2009): Can
choice feeding improve performances and N-retention of broilers compared to a
standard three-phase feeding schedule. British Poultry Science, Sep 50 (5): 573 -
582.
ELWINGER K., SVENSSON L. (1996): Effect of dietary protein content litterand
drinker type on ammonia emission from broiler houses. Journal of Agricultural
Engineering Research 64: 197-208.
FERGUSON N.S., GATES R.S., TARABA J.L., CANTOR A.H, PESCATORE
A.J., FORD M.J, BURNHAM D.J. (1997): The effect of dietary crude protein on
growth, ammonia concentration, and litter composition in broilers. Poultry Science
77, 1481-1487.
FERGUSON N.S., GATES R.S., TARABA J.L., CANTOR A.H., PESCATORE
A.J., STRAW M.L., FORD M.J., BURNHAM DJ. (1998): The effect of dietary
protein and phosphorus on ammonia concentration and litter composition in
broilers. Poultry Science, Vol. 77 (8): 1085-1093
GOMES G.A, ARAUJO L.F., PREZZI J.A., BARBOSA L.C.G.S., SAVIETTO D.
AND FILHO CREN A.W. (2006): Period of feeding a pre starter diet on
performance up to 42 days of broilers of different body weights at housing. book of
Abstrats, XII European Poultry Conference, Italy, 298-299.
MADRIGAL S.A., WATKINS S.E., WALDROUP P.W. (1994): Feeding
Programs Designed to Modify Early Growth Rates in Male Broilers Grown to 56
Days of Age. Journal of Applied Poultry Research 3:319-326.
NAMROUD N.F., SHIVAZAD M., ZAGHARI M. (2008): Effects of Fortifying
Low Crude Protein Diet with Crystalline Amino Acids on Performance, Blood,
Ammonia level, and Excreta Characteristics of Broiler chicks. Poultry Science 87:
2250-2258.
NIKOLOVA N., PAVLOVSKI Z., MILOŠEVIĆ N., PERIĆ L (2007): Uticaj
različitog nivoa energije i proteina u hrani na proizvodne parametre brojlerskih
pilića dva genotipa. Biotechnology in Animal Husbandry, Vol. 23, br. 5-6-1, str.
551-557.
PETRIČEVIĆ V., PAVLOVSKI Z., ŠKRBIĆ Z., LUKIĆ M. (2011): The effect of
genotype on production and slaughter properties of broiler chickens. Biotechnology
in Animal Husbandry 2011 Volume 27, Issue 2, Pages: 171-181.
423

424
N. Tolimir al.
POPE T., EMMERT J.L. (2001): Phase –feeding supports maximum growth
performance of broiler chicks from forty –three to seventy-one days of age. Poultry
Science 80 (3): 345-52.
PRAVILNIK O METODAMA UZIMANJA UZORAKA I METODAMA
FIZIČKIH, HEMIJSKIH I MIKROBIOLOŠKIH ANALIZA STOČNE HRANE
(1987): Službeni list SFRJ, broj 15/87.
REZAEI, A. TEIMOURI J. POURREZA, H. SAYYAHZADEH and
P.W.WALDROUP (2006): Effect of diet dilution in the starter period on
performance and carcass characteristics of broiler chicks. Journal Central European
Agriculture, Volume 7, No. 1 (63-70).
SAKI A.A., MOMENI M., TABATABAEI M.M., AHMADI A., RAHMATI
M.M.H., HEMATI MATIN H.R., JANJAN A. (2010): Effect of Feeding Programs
on Broilers Cobb and Arbor Acres plus Performance. International Journal of
Poultry Science 9 (8): 795-800,
SALEH E.A., WATKINS S.E., WALDROUP A.L., WALDROUP P.W. (2004):
Effect of Dietary Nutrient Density on Performance and Carcass Quality of Male
Broilers Grown for Further Processing. International Journal of Poultry Science 3
(1): 1-10.
SALEH E.A., WATKINS S.E., WALDROUP P.W. (1997): Changing Time of
Feeding Starter, Grower and Finisher Diets for Broilers. 2. Birds grown to 2,2 kg.
Journal of Applied Poultry Research 6: 64 73.
SALEH, E.A., WARKINS S.E., AND WALDROUP AND W.P.(1996): Changing
time of feeding starter, grower, and finisher diets for broilers.1. Birds grown to
1kg. J.Appl.Poult.Res. 5:269-275
TOLIMIR N, PERIĆ L, MILOŠEVIĆ N, BOGDANOVIĆ V (2010): The effect of
multiphase nutrition on production performances of broilers. Biotechnology in
animal husbandry, Vol.26, 1-2, 83-91.
WARREN W.A. AND EMMERT J.L (2000): Efficacy of phase-feeding in
supporting growth performance of broiler chuck during the starter and finisher
phases. Poult.Sci. 79:764-770.
Received 16 May 2012; accepted for publication 21 July 2012

Biotechnology in Animal Husbandry 28 (3), p 425-431 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 637.05
DOI: 10.2298/BAH1203425D
EGG QUALITY OF JAPANESE QUAIL IN SERBIA
(Coturnix coturnix japonica)
M. Đukić Stojčić 1 , N. Milošević 1 , L. Perić 1 , I. Jajić 1 , N. Tolimir 2
1
Faculty of Agriculture, University of Novi Sad, Trg Dositeja Obradovića 8, 21000, Novi Sad,
Republic of Serbia
2
Institute for Science Application an Agriculture, Bulevar despota Stefana 68b, 11000, Belgrade,
Republic of Serbia
Corresponding autor: mirjana.djukicstojcic@stocarstvo.edu.rs
Original scientific paper
Abstract: In the last years Japanese quail (Conturnix coturnix japonica) is
becoming more popular as a source of meat and eggs. Information on egg quality
characteristics has been limited mostly to chicken eggs. The aim of this paper is to
enhance the knowledge on the quality of quail eggs. In this study external and
internal quality traits of quail eggs from three different commercial farms will be
presented. For egg mass, shape index, shell thickness, shell mass no significant
difference was found between farms. For yolk colour, yolk and albumen mass,
statistically significant differences were found between the three farms. The
difference in shell breaking strength between farms A and B was not significant
and significant differences were found between eggs from farm C. Yolk, albumen
and shell percentage were in the same relation as the mass of these parameters. The
worst albumen quality was recorded in eggs from farm A. The parameters yolk
colour, Haugh Unit and egg proportions (albumen, yolk and shell) do not differ
between chicken and quail eggs. On the other hand, parameters which differ are
egg mass (about five times smaller at quail eggs), shape index, shell breaking
strength and shell thickness, which was to be expected. The external and internal
egg quality traits of quail eggs from three farms in Serbia do not differ from the
results of quality traits from other countries. On the other hand, this investigation
contributes the development of science, because it includes some parameters,
which have so far not been published in literature by other researchers from this
area.
Key words: eggs, quail, internal and external quality traits
Introduction
In the last years Japanese quail (Conturnix coturnix japonica) is important as a
laboratory animal, due to its easy maintenance, early sexual maturity, shorter

426
M. Đukić Stojčić et al.
generation interval, high rate of egg production, but Japanese quail is also
becoming more popular as a source of meat and eggs (Punya Kumari, 2008). Eggs
of most species of birds may have similarities in nutritional composition and
potential food usage, however, information on egg quality characteristics and
utilization of egg for food and other purposes have been limited mostly to chicken
eggs. Chicken egg has been very well studied for its quality as well as for its
composition, however such information are not so abundantly documented in other
poultry species (Dudusola, 2010). Among many quality characteristics, external
factors such as cleanness, freshness, egg weight and shell quality are important for
consumer´s acceptability of shell eggs, and interior characteristics such as yolk
index, albumen index, proportions of egg components and chemical composition
are also important for egg production industry (Song, 2000). Information on egg
quality characteristics has been limited mostly to chicken eggs.
Because of the growing interest in consumption of quail eggs in our country, and
due to the lack of recent investigations in this direction, the aim of this paper is to
enhance the knowledge on the quality of quail eggs, and to show the quality of
quail eggs in our surrounding. In this study external and internal quality traits of
quail eggs from three different commercial farms will be presented.
Materials and methods
The experimental material comprised eggs of Japanese quail (Coturnix
coturnix japonica) of laying type in their first year of production (between 25 and
30 weeks of age), derived from three commercial farms near Novi Sad. The quails
in all three farms are kept in battery-cages under same conditions. The quails were
fed ad libitum with the same complete commercial diets which can be found on the
market, which is the reason why these three farms were chosen. The investigation
of egg quality traits was carried out in the Department of Animal Science of the
Faculty of Agriculture in Novi Sad. Examination of egg quality parameters was
carried out on the random sample of 20 eggs per producer, one day after they have
been collected from the farms. The following egg quality traits (external and
internal) were assessed:
Egg weight (g), yolk weight (g), albumen weight (g), and shell weight (g) were
measured with analytic scale with 0.01 g accuracy.
To determine the proportions of egg parts, each egg was carefully broken and
shell separated. Eggs shell (not dried) was weighed and the relative weight
calculated by relating the shell weight to the weight of the egg. An egg separator
was used to separate the yolk from the albumen. Relative yolk weight was
calculated in percentages by relating the yolk weight measured to the nearest gram
to the whole weight of that particular egg and multiplied by 100. The albumen
weight was calculated by subtracting the yolk and shell weights from the whole

Egg quality of japanese quail ...
egg weight. The albumen weight relative to the individual egg weight was
calculated.
Yolk index (%) was calculated according to the formula: Yolk index = yolk
height (mm) x 100% / yolk width (mm).
Haugh units were calculated according to the formula (Haugh, 1937): HU =
100 log (H +7.57 -1,7 *M 0,37 ), H – average thick white height (mm), W – egg
weight (g).
Shell breaking force was measured by an Egg Force Reader (Orka Food
Technology Ltd, Israel). The stand of the device was modified for measuring quail
eggs.
Egg yolk colour was determined according to Roche yolk colour fan
(Vuilleumier, 1969).
Eggshell thickness (mm) was measured together with shell membranes at the
equatorial part of the egg using a micrometer screw.
Based on obtained data, statistical analysis was done using ANOVA and
Duncan post-hoc test (STATISTICA 8, Stat Soft Inc, 2007).
Results and Discussion
Table 1 presents results of egg quality traits. No significant differences were
found in the egg weight and shape index between eggs derived from three
producers.
The difference in shell breaking strength between farms A (1.72 kg) and B
(1.63 kg) was not significant. However, a significant difference was found between
eggs from farm C (1.05 kg) compared to eggs from farms A and B (P

428
M. Đukić Stojčić et al.
Table 1. Average values and standard deviation of external and internal egg quality traits of
quail and average values of chicken eggs quality traits
Producer
Quail eggs Hen eggs *
Parameters
A B C
X Sd X Sd X Sd
Egg weight (g) 11,52 1,03 12,30 0,59 12,18 1,00 55 -- 70
Shape index (%) 77,37 2,43 77,51 3,79 75,11 2,89 61 -- 86
Breaking strenght (kg) 1,72 a
0,33 1,63 a
0,38 1,05 b
0,50 4 --5
Shell thickness (mm) 0,196 1,780 0,201 1,610 0,186 1,350 0,32 - 0,35
Shell weight (g) 1,73 0,20 1,80 0,26 1,63 0,17 8 - 12 % from egg
Yolk weight (g) 3,72 b
Albumen weight (g) 6,07 c
Yolk colour (Roche) 13,6 a
Haugh Unit 83,65 b
Egg proportions (%)
Yolk 30,00 b
0,42 3,42 b
0,67 7,08 a
0,97 7,6 c
3,27 86,1 a
3,34 27,71 c
Albumen 55,15 b 4,37 57,67 a
0,49 4,15 a
0,38 6,4 b
2,32 10,9 b
3,02 87,93 a
2,85 32,28 a
4,16 52,61 c
0,32
0,74
2,18 1 -- 15
3,68 > 70
1,95 31
2,98 58
Shell 14,85 1,72 14,62 1,78 15,09 1,72 11
a-c Values within rows with no common superscript are significantly different (P

Egg quality of japanese quail ...
In this study, the average values that are determined are similar to the results
reported by Nazligul et al. (2001), Ozcelik et al. (2002), Kul and Seker (2004),
Nowaczewski et al. (2010) but different from results from Odunsi et al. (2007),
Ipek et al. (2007), Punya Kumari et al. (2008), Dudusola (2009, 2010).
The mean egg weight in this experiment was similar to the results that were
reported by Dudusola (2009, 2010). Nowaczewski et al. (2010), who analyzed egg
weight changes according to age of the experimental birds, found that the value of
this trait in week 25 (10.91 g) was smaller than in week 9 (11.33 g), but about 1 g
higher than in results obtained by Odunsi et al. (2007), who evaluated three protein
sources in the diets of growing and laying Japanese quails. The investigation from
Punya Kumari et al. (2008) showed that mean egg weight from quail in 16 th week
of production was 13.71 which is more than 1 g higher compared to our results.
Shape index, shell thickness, albumen weight, yolk weigh, Haugh Units and
percentages of yolk and albumen were similar to the results reported by most
researches (Nazligul et al., 2001; Ozcelik et al., 2002; Kul and Seker, 2004; Punya
Kumari et al., 2008; Nowaczewski et al. 2010).
The value of shell breaking strength of quail eggs was between 1.05 and 1.72. This
value was not compared to the results of other authors because these authors did
not report the results of breaking strength. Generally it can be stated that the
breaking strength is smaller compared to chicken eggs, which can as well be
expected since the quail eggs have smaller weight and eggshell thickness.
The shell weight and percentage of shell in our investigation was bigger compared
to the results of other authors (Odunsi et al., 2007; Ipek et al., 2007; Dudusola
2009, 2010, Nowaczewski et al. 2010), because the shell weight was measured
directly after the shell was broken and separated, and the mentioned authors
measured it after drying.
In results from Odunsi et al. (2007) the yolk colour was lighter and in range
between 1.2 and 1.5 points (Roche). In investigation from Punya Kumari et al.
(2008) the average yolk colour was 5.37, which is as well lighter than in our
investigations, where the yolk colour was not uniform, and in the range between
7.6 and 13.6.
Conclusion
Generally it can be concluded that the external and internal egg quality traits of
quail eggs from three farms in Serbia do not differ from the results of quality traits
from other countries. On the other hand, this investigation contributes the
development of science, because it includes some parameters, which have so far
not been published in literature by other researchers from this area. Further
investigations in the field of chemical composition are needed.
429

430
Acknowledgment
M. Đukić Stojčić et al.
Research was financed by the Ministry of Education, Science and Tehnological
Development, Republic of Serbia, project TR 31033.
KVALITET JAJA JAPANSKE REPELICE(Coturnix
coturnix japonica)
M. Đukić Stojčić, N. Milošević, L. Perić, I. Jajić, N. Tolimir
Rezime
Poslednjih godina japanska prepelica (Coturnix coturnix japonica) postaje sve više
popularna kao izvor mesa i jaja. Informacije o karakteristikama kvaliteta jaja su
uglavnom bila vezana za kokošija jaja. Cilj ovoga rada je da unapredi znanje o
kvalitetu jaja prepelica. Predstavljeni su podaci o spoljašnjem i unutrašnjem kvalitetu
prepeličjih jaja sa tri različite komercijalne farme. Parametari težina jaja, indeks
oblika, debljina ljuske, težina ljuske se statistički značajno ne razlikuju između tri
farme. Boja žumanca, težina žumana i belanca statistički su se razlikovale između sve
tri farme. Nije bilo statistčki značajne razlike u sili loma između farmi A i B dok su se
jaja sa farme C statistički bitno razlikovala u sili loma od fame A i B. Najlošiji kvalitet
belanca zabeležen je u jajima sa farme A. Parametri boja žumanca, Hogove jedinice i
procentualni udeo pojedinih delova jaja (belance, žumance i ljuska) se ne razlikuju
bitno između kokošijih i prepeličijih jaja. Takođe hemijski sastav prepeličijih i
kokošijih jaja je veoma sličan. Sa druge strane parametri koji se razlikuju a za koje je
bilo i očekivati da će se razlikovati između kokošijih i prepeličijih jaja su težina jaja (
oko pet puta manja masa jaja prepelica u odnosu na kokošija jaja), indeks oblika, sila
loma i debljina ljuske. Spoljašnji i unutrašnji kvalitet prepeličijih jaja iz tri različite
farme u Srbiji se ne razlikuje od rezultata o kvalitetu prepeličijih jaja iz drugih
zemalja. Ovo istraživenje doprinosi razvoju nauke o kvalitetu prepeličijih jaja jer
obuhvata i neke parametre koji do sada nisu bili objavljeni u literaturi od strane drugih
istraživača iz ove oblasti.
References
DUDUSOLA, I. O. (2010): Comparative evaluation of internal and external
qualities of eggs from quail and guinea fowl. International Research Journal of
Plant Science, 1(5) 112-115.
DUDUSOLA, I. O. (2009): Effects of Storage Methods and Length of Storage on
some Quality Parameters of Japanese Quail Eggs. Tropicultura, 27 ( 1) 45-48.

Egg quality of japanese quail ...
HAUGH, R.R. (1937): The Haugh unit for measuring egg quality. U.S. Egg
Poultry Magazine, 43 552-553 and 572-573.
IPEK, A., CANBOLAT, O., KARABULUT, A. (2007): The Effect of Vitamin E
and Vitamin C on the Performance of Japanese Quails (Coturnix Coturnix
Japonica) Reared under Heat Stress during Growth and Egg Production Period.
Asian-Aust. J. Anim. 20 (2 ) 252 – 256.
KUL S., SEKER, I. (2004): Phenotypic Correlations Between Some External and
Internal Egg Quality Traits in the Japanese Quail (Coturnix coturnix japonica).
International Journal of Poultry Science, 3 (6) 400-405.
MILOŠEVIĆ, N. and PERIĆ, L. (2011): Tehnologija živinarske proizvodnje.
Univerzitet u Novom Sadu, Poljoprivredni fakultet, Novi Sad, pp. 33-42.
NAZLIGUL, A., BARDAKCIOGLU, H.E., TURKYILMAZ, N.K., ORAL, D.C.
(2001): The effect of cage density on egg weight, egg prodcution and feed
consumpstion in Japanese quails. J. Fac. Vet. Med. Univ. 27 (2) 429-438.
NOWACZEWSKI, S., KONTECKA, H., ROSIÑSKI,A., KOBERLING, S.,
KORONOWSKI, P. (2010): Egg Quality of Japanese Quail Depends on Layer Age
and Storage Time. Folia biologica, 58 3-4.
ODUNSI, A.A., ROTIMI, A.A., AMAO, E.A. (2007): Effect of Different
Vegetable Protein Sources on Growth and Laying Performance of Japanese Quails
(Coturnix Coturnix Japonica) in a Derived Savannah Zone of Nigeria. World
Applied Sciences Journal, 3 (5) 567-571.
OZCELIK, M. (2002): The phenotypic correlations among some external and
internal quality characteristics in Japanese quail eggs. Vet. J. Ankara Univ., 49 67-72.
PAVLOVSKI, Z., HOPIĆ, S. and LUKIĆ, M. (2001): Housing systems for layers
and egg quality. Biotechnology for Animal Science, 17, 197-200.
PUNYA KUMARIL, B., RAMESH GUPTA, B., GNANA PRAKASH, M.,
RAJASEKHAR REDDY, A. (2008): A study of egg quality traits in Japanese
quails. J. Veterinary & Animal Sciences 4 (6) 227-231.
SONG, K. T., CHOI, S. H., OH, H. R. (2000): A comparison of Egg Quality of
Pheasant, Chukar, Quail and Guinea Fowl. Asian-Aus. J. Anim. Sci., 13 (7) 986-
990.
VUILLEUMIER, J.P. (1969): The ‘Roche Yolk Colour Fan’-An Instrument for
Measuring Yolk Colour. Poultry Science, 48: 767-779.
Received 16 May 2012; accepted for publication 11 August 2012
431

Biotechnology in Animal Husbandry 28 (3), p 433-439 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.084.42
DOI: 10.2298/BAH1203433U
THE EFFECT OF FEED WETTING AND
FERMENTATION ON THE PERFORMANCE OF
BROILER CHICK
E. N. Uchewa, P. N. Onu
Department of Animal Science, Ebonyi State University, P.M.B 053, Abakaliki, Ebonyi State,Nigeria
Corresponding author: nnenwamazi@yahoo.com
Abstract:An experiment was conducted to evaluate the effect of feed
wetting and fermented feed on the performance of broiler chicks. Four
experimental diets were used in the study. Diet 1, was a conventional commercial
broiler starter feed. Diet 2 was a commercial broiler starter feed in a 1:1.2 dilution
with water. Diet 3, was a water diluted commercial feed inoculated with
Bactocell and fermented for 24 hours at 35 o C in an incubator. Diet 4 was
fermented feed add with 6% copper sulphate solution at the rate of 1ml to 10g of
feed, to then incubated for 24 hours. 180 day old Anak 2000 broiler chicks were
randomly assigned to the four experimental diets in a completely randomized
design (CRD). Each treatment group was replicated four times with 15 birds per
replicate. Results showed that feed intake and weight gain were significantly higher
(P

434
E. N. Uchewa et al.
an average Nigeria consumes about 53.8g protein out of which animal protein
contributed only 8.4g. As a result of this, efforts have been geared towards
alleviating the animal protein deficit plaguing Nigerians, through improved animal
production.
Nutrition is by far one of the greatest factors influencing the productivity
of farm animals. This is because feeding accounts for over 70 % of the total cost of
producing intensively kept animals (Esonu, 2001). However, in a bid to reduce the
cost of feeding, farmers resort to feeding poor quality feeds to farm animals. This
has resulted lately to economic losses in poultry production in developing countries
(Aletor, 1986). Hence the need to focus attention on strategies for maintaining the
quality of feeds at reduced to ensure improved utilization and performance of
poultry.
In Nigeria, commercial poultry birds are usually fed dry mash. Wet feeding
involves the addition of water to dry poultry mash before feeding. Earlier
comparison between the use of dry and wet mash (Yalda and Forbes, 1995; Kutlu
et al., 1997) showed significant increase in feed intake, body weight gain and feed
efficiency of birds when the feed was mixed with up to twice the weight of water to
give a porridge-like consistency. Studies by Awojobi and Meshioye (2001) and
Ogbonna et al. (2001) showed that wet mash was more beneficial than dry mash in
poultry feeding. Similar observations have been reported in growing pigs (Brooks
and Carpenter, 1990; Rayner and Miller, 1990). Meanwhile, fermentation of feed
samples has been reported to alter both physio-chemical and microbiological
properties of feed with improved performance of animals (Moran, 2001). Newman
and Newman (1985) reported that fungal fermentation of barley with Rhizopus
oligosporus improved the performance of broiler chickens. Adejimni et al. (2003)
indicated that fermented cocoa husk improved weight gain of rabbits. However,
there is still little of information on the feeding value of fermented liquid feed to
broiler chickens.
The present study was therefore embarked upon to evaluate the
performance of broiler chickens fed fermented and unfermented commercial
broiler feed.
Materials and Methods
The experiment was conducted at the Teaching and Research Farm
(Poultry Unit), Department of Animal Science, Ebonyi State University, Abakaliki.
The experimental protocol was approved by the Animal Care and Use Committee
of the Institution.
Preparation of the experimental diets. A batch of a commercial broiler
starter ration was purchased and calculated per group by subtracting the initial
body used for the preparation of the test diets. Four experimental diets were

The effect of feed wetting ...
prepared. Diet 1, (control) was a dry commercial broiler starter feed. Diet 2 was a
dry feed mixed with water in a ratio of 1:1.2g of water solution while diet 3 was the
water mixed feed inoculated with actocell to give a final concentration of 7
log10 CFU/ml liquid feed. Diet 4 was an equal part of diet 3 mixed with 6% copper
sulphate solution in a ratio of 10:1. Both diets 3 and 4 were then kept in the
incubator to ferment for 24 hours at 35 o C. The proximate composition of the
commercial diet is shown in Table 1.
Table 1: Proximate composition of the commercial broiler starter diet used in the study
Ingredients Inclusion (%)
Crude protein 21
Ether extract 4.0
Crude fibre 3.8
Total Ash 5.5
Methionine 0.52
Lysine 0.92
Available phosphorus 0.45
Vitamin A 15000IU Kg -1
Vitamin D3 5000IUKg -1
Metabolizable energy 14MJ Kg -1
Experimental animals and procedure. A total of one hundred and eighty
(180) Anak 2000 broiler chicks procured from a commercial hatchery were used
for the experiment. Prior to the commencement of the experiment, the birds were
fed a commercial broiler starter for one week adaptation period. On the eight day
they were divided into four treatment groups of birds each and each group
randomly assigned to an experimental diet in a completely randomized design
(CRD). Each treatment was further subdivided into three replicates of 15 per
replicate were housed in pens compartment measuring 3m x 3m electric bulbs.
Heat was provided to the birds using Routine poultry management practices which
included daily inspection of the birds for symptoms of diseases, mortality, cleaning
of troughs and supply of feed and fresh water were maintained. The experiment
lasted for 28 days. The care and management of the experimental animals has been
carried out by the procedure according to the recommended guidelines of
Federation of Animal Science Societies (1999).
Data collection and analysis. A weighed quantity of feed was served to
the birds. Left over feed was collected per group every morning, weighed and
recorded. From this the daily feed intake of each replicate group was determined by
difference between the quantity of feed served and the leftover. Weighing of the
birds was done weekly on individual basis and in the morning when their crops are
virtually empty. At the end of the trial, the body weight gain was calculated per
group by subtracting the initial body weight from the final weight. The daily body
weight gain was determined by dividing the body weight gain with the number of
435

436
E. N. Uchewa et al.
days the experiment lasted. The feed conversion ratio (FCR) was computed by
dividing the average daily feed intake by the average daily weight day.
Carcass evaluation. At the end of the feeding trial, five birds were
randomly selected from each experimental group, weighed, slaughtered,
eviscerated, dressed and cut into individual parts for carcass evaluation. The
various organs were dissected and blotted free of blood and their weights taken on
a G&G Electronic Scale and Mettler P 2000 weighing balance and later expressed
as percentage of the body weights. Data collected were subjected to a one-way
analysis of variance (Steel and Torrie, 1980). Differences between the treatment
means were separated using Duncan’s New Multiple Range Test as outlined by Obi
(2002).
Results and Discussion
The data on the performance of the broiler chicks fed the experimental
diets are shown on Table 2. There were significant (P < 0.05) differences in all the
parameter measured among birds fed the control and liquid feeds respectively.
Feed intake and weight gain were significantly higher (P < 0.05) in birds fed
fermented feed and non-fermented liquid diets. These findings were in agreement
with that reported by Yalda and Forbes (1995) and Awojobi and Meshioye (2001).
The higher feed intake of birds fed liquid feeds may be attributed to wetness of the
feed which may have increased the rate of passage of the feed through the gastro
intestinal tract. Interestingly, the higher feed intake resulted to higher body weight
gain, thus indicating more efficient utilization of the feed by the birds.
Birds on dry feed consumed significantly (P < 0.05) more water than those
on liquid feed. Thus suggesting that a remarkable part of the water requirement of
birds on liquid feeds were met by the wet feed (Yalda and Forbes, 1995; Awojobi
and Meshioye, 2001; Ogbonna et al., 2001).
Table 2: Performance of broiler chicks fed the experimental diets
Treatments
Parameters. T 1 T 2 T 3 T 4 SEM
Initial body weight. (g) 48.3 50.1 49.8 48.4
Final body weight (g) 1017.77 c 1229 a 1231.0 a 1195.83 b 5.57
Body weight gain (g) 969.47 c 1179.10 a 1181.28 a 1147.43 b 5.36
Daily body weight gain (g) 23.08 b 28.07 a 28.12 a 27.32 a 2.11
Total feed intake (g) 2843.62 b 3208.98 a 3285.64 a 3232.82 a 8.79
Daily feed intake (g) 67.71 b 76.40 a 78.23 a 76.87 a 2.98
Water intake (ml) 321.45 a 245.1 226.78 b 232.26 a 3.78
Feed conversion ratio 2.93 b 2.72 a 2.78 a 2.81 a 0.01
ab = means on the same row different superscripts are significantly different (P < 0.05). SEM =
standard error of means. T1 =conventional dry broiler starter diet, T2= unfermented liquid diet T3=
fermented liquid diet, T4 = fermented liquid feed with copper sulphate added.

The effect of feed wetting ...
Birds receiving liquid feed (fermented and non-fermented) showed
significantly (P < 0.05) superior feed conversion ratio over the control group. The
better feed conversion ratio is not surprising since it was discovered that the birds
spent less time feeding on the wet feed and so expended less energy than those fed
dry feed (Savory, 1974). Liquid diets compared favourably (P > 0.05) in all the
response parameters measured. In spite of the non-significant (P > 0.05) difference
in the performance of the birds fed fermented and non-fermented liquid diets; there
were tendencies for more efficient utilization of the nutrients in non-fermented
liquid diets. This is seen in the marginal improvement of 2.16% of feed conversion
ratio in favour of non-fermented liquid feed.
Table 3: Carcass characteristics of broiler chicks fed the experimental diets.
Treatments
Parameters T1 T2 T3 T4 SEM
Final body weight (g) 1017.77 b 1229 a 1231.0 a 1195.83 a 5.57
Dressed weight (%) 86.08 86.53 86.28 86.14 0.05
Thigh 1 13.96 14.10 14.00 14.02 0.14
Shank 1 7.08 7.12 7.12 7.09 0.24
Wings 1 10.45 10.60 10.65 10.56 0.36
Breast 1 14.61 14.72 14.65 14.62 1.98
Gizzard 2 3.30 3.81 3.79 3.58 0.65
Heart 2 2.09 2.19 2.22 2.10 0.11
Liver 2 2.54 2.67 2.82 2.58 0.22
1 value expressed as gram per kilogram body weight, 2 values expressed as a percentage of body
weight, T1 =conventional dry broiler starter diet, T2= unfermented liquid diet T3= fermented liquid
diet, T4 = fermented liquid feed with copper sulphate added.
Data on the carcass characteristics of broiler chicks as influenced by the
test diets (Table 3) indicate that average live weight, dressed weight, wings, breast,
thigh, and shank were not significantly (P>0.05) affected by the dietary treatments.
Dressed weights of birds were not significantly (P>0.05) influenced by the test
diets, but the trend showed that the highest dressed weight was with birds fed non
fermented liquid feed followed by fermented liquid feed. The relative organ
weights (organ weight to body weight ratios) of broiler chicks indicated that the
relative weights of liver, gizzard and heart were not significantly (P

438
E. N. Uchewa et al.
Uticaj kvašenja hrane i fermentacije na proizviodne rezultate
brojlerskih pilića
E. N. Uchewa i P. N. Onu
Rezime
Cilj eksperimenta je bio da se proceni uticaj kvašenja hrane i fermentisane hrane na
proizvodne performanse brojlerskih pilića. U ogledu su korišćena četiri obroka.
Obrok 1, je bila konvencionalna, komercijalna starter hrana za brojlere. Obrok 2 je
bila komercijalna starter smeša razređena u vodi u 1:1.2. Obrok 3 je bila
komercijalna hrana razređena u vodi sa inokulantom Bactocell, fermentisana u
trajanju od 24 sata na 35 o c u inkubatoru. Obrok 4 je sadržavao feremntisanu hranu
gde je dodat 6% rastvor bakar sulfata u 1ml na 10g hrane i inkubiran u trajanju od
24 sata. Brojlerski pilići Anak uzrasta od 180 dana su metodom slučajnog izbora
podeljeni u 4 eksperimentalne grupe 180 дана старе Анак 2000 бројлерски
пилићи су насумично додељују у четири експерименталнa tretmana
(completely randomized design - CRD). Za svaki tretman je bilo po četiri
ponavljanja - 15 grla po ponavljanju. Rezultati su pokazali da je unos hrane i
prirast bili signifikantno viši (P

The effect of feed wetting ...
ALETOR V.A. (1986): Some agro-industrial by-products and wastes in livestock
feeding. A review of prospects and problems. World Review in Animal Production,
22: 36-41.
AWOJOBI H.A., MESHIOYE O.O. (2001): A comparison of wet and dry mash
feeding for broiler finisher during wet season in the tropics. Nig. J. Anim. Prod., 2,
143-146.
BROOKS P.H., CARPENTER J.L. (1990): The water requirement of growing
finishing pigs–theoretical and practical considerations. In: HARESIGN W. AND
COLE D.J.A. (ed) Recent Advances in Animal Nutrition, 115-136.
ESONU B.O. (2001): Animal nutrition and feeding: A functional approach.
Rukzed and Rucksons Association, Owerri, Nigeria.
FAO (1984): FAO production year book. Food and Agricultural Organization of
the United Nation, Rome.
FEDERATION OF ANIMAL SCIENCE SOCIETIES (1999): Guide for the care
and use of Agricultural Animals in Agricultural Research and Teaching 1st
revised edn. Savoy, IL.
KUTLU H.R., GORGULU M., DEMIR E., OZTURKCAN O. (1997): Effect of
wet feeding on performance and carcass composition of broiler chicks reared at
high ambient temperature. Proceedings of 10th Europe. Symp. on Poultry
Nutrition. Antalya, Turkey, 252-253.
MORAN C.A. (2001): Development and benefits of liquid diets for newly weaned
pigs. Ph.D Thesis, University of Plymouth, USA.
NEWMAN R.K., NEWMAN C.W. (1985): Effect of fungal fermentation and other
treatment on nutritional value of waxy barley fed to chicks. Poult. Sci., 64: 1514-
1518.
OBI I.U. (2002): Statistical method of detecting differences between treatment
means and research mythology issue in laboratory and field experiments. A P
Express publishing Coy. Ltd. Nsukka, Nigeria.
OGBONNA J.U., OGUNDOLA F.I. AND OREDEIN A.O. (2001): Effect of wet
feed on cockerel chicken performance. Nig. J. Anim. Prod., 1, 52-55.
RAYNER D.V. AND MILLER S. (1990): Cholecystokinin effects on wet and dry
meals in pigs. Proceedings of the Nutrition Society (British), 49, 22-26.
SAVORY C.J. (1974): Growth and behaviour of chickens fed pellets or mash. Br.
Poult. Sci., 15, 281-286.
STEEL R.G. AND TORRIE J.H. (1980): Principles and procedure of statistics. A
biometrical approach. 2 nd ed. McGraw – Hill Book. Co. Inc. N.Y.,USA.
YALDA A.Y. AND FORBES J.M. (1995): Effect of wet feeding on the growth of
ducks. Br. Poult. Sci., 36, 878-879.
Received 15 December 2011; accepted for publication 28 june 2012
439

Biotechnology in Animal Husbandry 28 (3), p 441-453 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.087.7
DOI: 10.2298/BAH1203441S
ESTIMATION OF THE RELATIVE BIOAVAILABILITY
OF SEVERAL ZINC SOURCES FOR BROILERS WHEN
FED A CONVENTIONAL DIET
M. Sahraei 1 , H. Janmmohamadi 2 , A. Taghizadeh 2 , G. Ali Moghadam 2 ,
S. A. Rafat 2
1 PhD, Student of Poultry Nutrition, Tabriz University,Tabriz, Iran,5166616471
2 Prof. of Depatrment of Animal Science,Tabrize University,Tabriz, Iran, 5166616471
Corresponding Author: m.sahraei2009@gmail.com
Original scientific paper
Abstract: An experiment was conducted with 312 day-old male broiler
chicks in grower phase(8-28d) to estimate the biological availability of four
sources Zinc (Zn), Zinc sulfate(ZnSO4.H2O), two sources of Zinc oxide(ZnO FG1
and ZnO FG2) and Bioplex Zn. Zinc sulfate (ZnSO4.H2O) was used as the standard
in the bioavailability assay. Chicks were allotted randomly to 13 dietary treatments
with 6 birds per replicate and 4 replicates per treatment, that included an
unsupplemented corn-soybean meal basal diet (25.50 mg of Zn/kg of DM), or the
basal diet supplemented with 100, 150 or 200 mg/kg of DM as either ZnSO4.H2O
(33 % Zn), Zinc oxide FG1(72%Zn), Zinc oxide FG2 (75%Zn) or Bioplex Zn
(15% Zn). Dietary Zn level and source had no effect (P>0.05) of feed intake or
body weight gain of chicks during first and second weeks of experimental periods,
but feed conversion ratio in the first and second week and feed intake, body weight
gain or feed conversion ratio in third week and total experimental periods were
significant difference between treatments (P

442
M. Sahraei et al.
Because many natural feed ingredients are marginally deficient in Zn (Falchuk and
Vallee, 1985; Kaim and Schwederski, 1994). The current (NRC,1994) zinc
requirement for broilers is 40 mg/kg, that based upon research conducted more
than 40 yr ago with animals of markedly different productive potential than those
that exist today, also this requirement is based on only a few research reports, most
of which were carried out using purified or semipurified diets with growth as the
only requirement criterion (Roberson and Schaible, 1958; Zeigler et al., 1961;
Emmert and Baker, 1995). Dietary Supplementation by inorganic trace minerals
have traditionally provided with sufficient amounts of each mineral to support
normal growth, health, and reproduction. However, genetic improvements
continually change the commercial broiler strains, and nutritionists start to question
if currently used trace mineral levels and sources will be suitable in the future when
feeding these faster-growing and highly productive birds for meat and eggs (Nollet
et al., 2008). Considerable confusion exists concerning bioavailability of Zn in
various Zn supplements for chickens and pigs. Using weight gain as a response
variable, Edwards (1959) reported that Zn from analytical grade (AG) and
technical grade ZnO as well as Zn metal powder was 100% bioavailable for young
chicks relative to AG ZnSO4.7H2O. The next year, Roberson and Schaible (1960),
also using chick weight gain as the response variable, reported that ZnO was as
bioavailable as ZnSO4, but they did not clearly identify either compound. Miller et
al. (1981) compared AG ZnO to Zn metal dust in pigs fed a corn-soybean meal
diet. Wedekind and Baker (1990) reported relative bioavailability (RBV) values for
chicks fed feed-grade (FG) Waelz-processed ZnO of 61% (weight gain) and 44%
(total tibia Zn) compared with FG ZnSO4.H2O (100%). Wedekind et al. (1994)
concluded, based on bone Zn accumulation in pigs, that FG ZnO was only
approximately 68% as bioavailable as FG ZnSO4.H2O. Edwards et al. (1998)
evaluated two byproducts of the galvanizing industry, Fe-ZnSO4.H2Oand Zn-
FeSO4 .H2O, and found both to be as bioavailable, based on weight gain and total
tibia Zn, as FG ZnSO4.H2O. Many experiments have been conducted during the
last 50 yr to estimate the bioavailability of Zn in supplemental sources and dietary
ingredients; however, there are considerable confusion concerning bioavailability
of this nutritionally trace element in various supplements for chickens and pigs.
Using weight gain as a response variable, Edwards (1959) reported that Zn from
analytical grade and technical grade ZnO as well as Zn metal powder was 100%
bioavailable for young chicks relative to analytical grade ZnSO4.7H2O. Some
researchers (Spears, 1989;Wedekind et al., 1992) have reported greater bio-efficacy
for organic Zn sources than that observed for inorganic forms, including Zn oxide
and Zn sulfate; consequently, organic forms of the trace element have been used
with increasing pattern by the feed industry. An enhanced bioavailability of a
mineral source could reduce the amount of a mineral that is added to a diet to meet
mineral nutritional requirements, which would reduce the amount of mineral
excreted by birds (Cheng et al., 1998). The use of organic complexes of trace
element, such as Zn-lysine (ZnLys) and Zn methionine (ZnMet), has received more

Estimation of the relative bioavailability ...
attention because of The data reported by Wedekind et al. (1992) indicated that, for
chicks fed corn-soybean meal diets, Zn from a Zn methionine complex was 206%
bioavailable relative to a ZnSO4.H2O standard (i.e. 100%), where ZnO provided
only 61% bioavailable Zn. There are conflicting data reported regarding the
relative efficacy of different organic versus inorganic Zn sources in enhancing
broiler performance. The research presented herein, therefore, aimed to compare
bio-efficacy of various inorganic and organic Zn compounds in the light of their
effects on the performance of broiler chicks fed corn-soybean meal diets.
Materials and Methods
Birds, treatments and managing program
This study was conducted in the poultry research farm of Tabriz
University (Tabriz, Iran). A total of 312 day-old Ross308 male broiler chicks were
randomly allotted to four pen replicates of six birds for each of 13 dietary
treatments such that each pen had a similar initial weight and weight distribution.
Table 1: Composition of the basal diets
Ingredients (%) (8-28d)
Corn 55.90
Corn Starch 0.05
Soybean meal 36.60
Soybean oil 3.60
Di-Calcium phosphate 1.50
Caco3 1.35
Salt 0.25
DL-methionine 0.15
L-lysine 0.10
Vitamin premix a 0.25
Zn free mineral premix b 0.25
Nutrient composition
Metabolizable energy (Kcal/kg) 3000
Crude protein(%) 21.00
Calcium 0.95
Available Phosphorus(%) 0.44
Lysine (%) 1.19
Methionine (%) 0.49
Zinc (mg/kg) 25.50
a:Supplied per kg diet: 11,025 I.U. vitamin A, 3,528 I.U. vitamin D3, 33 I.U. vitamin E, 0.91 mg; vitamin K, 2 mg
thiamin, 8 mg riboflavin, 55 mg niacin, 18 mg Ca pantothenate, 5 mg vitamin ;B6, 0.221 mg biotin, 1 mg folic
acid,478 mg choline, 28 μg vitamin B12
b:Zinc-free mineral premix. Provided per kilogram of diet: Mn (fromMnSO4 H2O), 60 mg; Fe (from
FeSO4 7H2O), 50 mg; Cu (from CuSO4 5H2O), 6 mg; I (from Ca (IO3)2 H2O), 1 mg; Se, 0.20 mg.
The basal corn-soybean meal diets (Table 1) containing 25.50 mg Zn/kg as
fed basis (by analysis) were formulated to meet or exceed nutritional requirements
443

444
M. Sahraei et al.
of broiler chicks (NRC, 1994) grower stages from 8-28 d. Dietary treatments
included the basal diet or basal diet supplemented with 100, 150, or 200 mg/kg
added Zn as feed-grade Zn sulfate , Zn oxide feed- grade1, Zn oxide feed- grade2
and Bioplex Zn (as organically Zn compounds). The basal diets were formulated
using Zn-free mineral premix, so that contained minimum amount of zinc. A single
batch of basal feed was mixed and divided into 13 aliquots according to the
experimental treatment, Each Zn source was mixed with cornstarch to the same
weight and mixed with each aliquot of the basal diet. All of the diets were
calculated to contain equal concentrations of methionine, lysine and other nutrients
except of Zn. Chicks were housed in the cage pens which placed at
thermostatically-controlled room. Chicks were maintained on a 24 h constant
lighting regimen and had free access to feed and tap water containing no detectable
Zn in all times. Feed and water were provided using plastic instruments to
minimize environmental Zn contamination.
Chemical analysis
Prior to formulating the diets, the ingredients and Zn sources used were
analyzed for crude protein, ether extract, crude fiber, and ash content according to
standard procedures (AOAC,1995). Zinc concentrations in Zn sources, diets and
water were determined by atomic absorption spectrophotometery (Perkin Elmer,
Precisely A Analyst 200, Absorption spectrophotometer).
Data collection
Body weight, body weight gain, feed conversion ratio and Zn intake were
recorded weekly and overall experimental period. Relative bioavailability values
were estimated by a slope ratio techniques from regression of weight gain on Zn
intake with ZnSO4.H2O as the standard source at 100% (Finney,1978, Littell et al.,
1997). Because feed intake differences among treatments could affect Zn intake,
regressions were calculated using dietary Zn intake (based on Zn assays of diets) as
the independent variable rather than added Zn concentrations (Wedekind et al.,
1992, Li et al.,2004).
Statistical analysis
Pen means data were subjected to ANOVA procedures of SAS, PC version
6.12 (1999), appropriate for completely randomized designs. Regression equations
for the ZnSO4.H2O standard curves were derived using Minitab 10. Weight gain
was regressed on supplemental Zn intake from ZnSO4.H2O, and Zn bioavailabilities
in the test sources of Zn were determined using standard-curve
methodology .Relative bioavailability estimates were calculated for each of the
four replicate pens that were fed each experimental Zn source. Duncan’s multiple
range tests (Duncan, 1955) was used to separate treatment means at p

Results and discussion
Estimation of the relative bioavailability ...
Chemical characteristics of mineral sources are presented in Tables 2. The
Zinc sources studied herein (zinc sulfate, Zinc oxide FG1, Zinc oxide FG2 and
Bioplex Zn) were determined to be containing 32, 72, 75 and 15% zinc as fed
basis, respectively.
Table 2. Characterization of zinc sources under investigation
Zn source a
Zn level, % b
Color
ZnSO 4.H 2O(FG) 33 White
ZnO(FG 1) 72 White
ZnO(FG 1) 75 White
Bioplex Zn c
a FG: feed grade.
b Determined from atomic absorption spectrophotometery.
c Supplied by Alltech Biotechnology
15 Yellow
The effects of source and supplemental zinc level on performance
parameters are shown in Table 3. The Zn source and concentration did not affect
Feed intake (FI) or weight gain (WG), in the first or second week and body weight
(BW) in 14 d,21d and 28d (P>0.05) but feed conversion ratio(FCR) in all
experimental periods, Feed intake (FI) or weight gain (WG) in the third week and
overall trial periods were, significantly (p

446
M. Sahraei et al.

Estimation of the relative bioavailability ...
As presented in Table 4, the relative biological availability values using
body weight gain as criteria were estimated to be 59, 99 or 45 for three levels of
Zinc oxide FG1, 64, 78 or 31 for three levels of Zinc oxide FG2 and 151, 200 or
147 for three levels of Bioplex Zn, respectively. The highest RBV of different Zinc
sources was observed in the 150 ppm levels of these sources when used in diets.
Table4. Estimated relative Zn bioavailability values of weight gain(g/21d) on dietary zinc
sources intake (mg/21d) (Zn from ZnSO4.H 2O set at 100%) a
Zinc source
Added
Zn,ppm
Regression
equation b
RBV(%) c
100 Y= 135+8.87X 100 0.86
Sulfate 150 Y=239+5.81X 100 0.93
200 Y=-79.7+8.16X 100 0.98
100 Y=154+5.25X 59 0.86
Oxide FG1 150 Y=-35+5.15X 99 0.91
200 Y=37.5+3.75X 45 0.99
100 Y=124+5.69X 64 0.91
Oxide FG2 150 Y=95+4.54X 78 0.94
200 Y=265+2.52X 31 0.53
100 Y= 37.3+13.4X 151 0.96
Bioplex 150 Y=55+11.6X 200 0.94
200 Y=-57.2+12X 147 0.98
aData are means of four pens of six male chicks fed the experimental diets for 21d (d 8 to d 28 posthatching);
average initial weight was 92 g.
bStandard curve for weight gain (Y, in g/21 d) regressed on supplemental Zn intake (X, in mg/21 d) from different
zinc sources.
cRelative bioavailability (RBV) was calculated from the standard curve regressions, setting RBV of Zn in the
ZnSO4.H2O standard at 100%.
Supplementation of Zn to the basal diet at graded levels had no significant
influence on body weight gain and feed intake at 3week of age (Table 3). This was
in agreement with the findings of Burrell et al. (2004) who found that a practical
diet of maize-soybean meal containing 30 ppm of Zn was adequate to support
optimum performance during the initial 3 week of age. Similarly, others (Stahl, et
al., 1986) reported that a basal diet containing 37 ppm of Zn was optimum for
realizing good growth in chicks and additional supplementation had no added
advantage. In another study with male broiler chicks, little effect was found on
body weight, feed efficiency, or livability with supplementation of Zn up to 6 week
of age because the basal diet contained 44 ppm of Zn. The present study also
indicated that the Zn content available in cornsoybean diet (25.50 ppm) was
R 2
447

448
M. Sahraei et al.
adequate to sustain growth and other related parameters at par with those
supplemented with Zn up to 4 week of age. Obviously, this level was lower than
that recommended by NRC (1994) (40 ppm) for broiler chicks. The absence of any
difference in performance between groups fed diets with or without supplemental
Zn could be due to slower rate of Zn utilization, necessitating no further
replenishment in diets (Collins and Moran, 1999). The lack of increase in feed
intake or body weight gain with added Zn in this phase indicates that the amount of
the element in the non-supplemented basal diet was adequate for growth specially
until 21 day of life despite of the fact that NRC (1994) suggested that 40 mg/kg as
the requirement for broiler chicks. The contribution of Zn from the remaining yolk
sac and easier permeability of the still developing gastrointestinal tract may have
contributed to this observation (Cao et al., 2002). Also the lack of responses in
feed intake and weight gain to added Zn levels up to 100 ppm, probably due to
increased synthesis of intestinal metallothionein. Zinc intake has been shown to
induce intestinal metallothionein synthesis (Sandoval et al., 1997). Increased
synthesis of this zinc binding protein is associated with reduced zinc absorption.
This protein will influence the regulation of Zn absorption and possibly the
response of broiler chicks to supplemental levels of zinc from different sources.
But the improvements effects of adding different zinc sources of FCR in week3
and total experimental periods indicated that the using of this supplement might be
suitable in practical diet of broilers in grower phase. Bioavailability of trace
minerals is defined as the proportion of the ingested element that is absorbed,
transported to its site of action, and converted to a physiologically active form
(Ammerman et al., 1995). Thus, bioavailability implies not only absorption but also
utilization of the mineral for a specific function. However, it is difficult to
quantitatively evaluate the actual utilization of an element with a response criterion
of sufficient sensitivity to determine statistical differences with a small population
of animals (Li et al., 2005). The old experiments designed to assay the
bioavailability of Zn usually used purified diets supplemented with low
concentrations of Zn and growth performance and tibia Zn concentration as the
response criteria (Edwards, 1959; Wedekind and Baker, 1990). But, the estimate
determined in a purified diet might not be applicable to practical diets because of
the absence of phytate and fiber (Wedekind et al., 1992). Therefore, the later
experiments were designed to use a corn-soybean meal diet with a short-term,
high-supplemental Zn to determine relative bioavailability based on weight gain as
a simple assay factor (Cao et al., 1996, 2000; Sandoval et al., 1997). The method
offers several advantages in assessing Zn bioavailability such as: minimizing the
effect of decreased feed intake and also the applicability of the results to
commercial conditions, the diet is less expensive than semi-purified or purified
diets, the diet more palatable, therefore enabling maximum genetic growth
potential and the extrapolative value of the results is good, because the
conventional soybean meal diet is like to that used in commercial conditions

Estimation of the relative bioavailability ...
(Wedekind et al.,1992). Based on those findings, a practical corn soybean meal diet
(25.50 mg of Zn/kg) and 4 supplemental Zn concentrations (0, 100, 150, 200 mg
Zn/kg) were chosen in our study. In this experimental design, finding a criterion
that responds in a linear manner to dietary added Zn intake is most difficult and
important. Weight gains is one method to measure utilization but is generally a
fairly unresponsive criterion for many mineral elements (Luo et al., 2007). In the
present study, bioavailability estimates of Zn from the tested sources were based on
weight gains (Table 4) the best relative bioavailability value (RBV) was observed
in 150 mg of Zn/kg of basal diet in different Zinc sources relative to Zinc sulfate
as a standards. Also three levels of the Bioplex Zn had higher RBV than other
Zinc sources. The lower bioavailability observed for Zn oxide sources compared to
Zn sulfate in the studies reported herein is in agreement with Wedekind and Baker
(1990). The feeding study performed by Leeson (2003) showed that the
bioavailability of proteinated forms of elements supplemented to the diet for
broilers was at least by 30% higher than that of inorganic forms of elements. The
greater bioefficacy of ZnMet and/or ZnLys relative to sulfate or oxide forms
suggests that the metabolism of these Zn complexes differs from metabolism of Zn
supplied by inorganic Zn sources. These results would suggest that the major
differences in Zn bioavailability among different Zn sources can be attributed to
differences in absorption, and that once absorbed, differences in Zn utilization
among sources reflect differences in net utilization. These differences in utilization
may reflect differences in endogenous Zn loss, which has been shown to increase
due to homeostatic mechanisms with increasing Zn absorption (Weigand and
Kirchgessner, 1980). In poultry and swine nutrition, it is difficult to avoid the
presence of phytates as they are the main storage forms of P in seeds. Diets based
on corn and soybean meal generally contain between 2.0 and 2.5 g phytic P / kg.
Zinc content in feed components from plant origin is positively correlated to the
phytic P content, with ~10 mg of Zn to 1 g phytic P (Revy et al., 2003). It is
suggested that Zn from organic sources is protected by the ligand from reacting
with feed antagonists, such as phytate to form insoluble complexes. Zinc from
organic sources is absorbed by the intestinal cells as an ion, or it is also suggested
that Zn is absorbed intact thanks to a second absorption pathway related to the
ligand. Also a review of Zn bio-availability data (Ledoux, 2005; Wedekind et al.,
1992) indicates that in most studies, organic mineral sources were at least as
available as the standard inorganic sources, and in some cases were more available.
One of the hypothesized reasons for increased bioavailability of organic minerals is
that this form of mineral is protected form unwanted interactions in the
gastrointestinal tract. Conversely, some reports have shown no influences of
complexing with an organic ligand (protein, methionine, or lysine) on mineral (Zn
and Mn) bioavailability (Aoyagi and Baker,1993; Pimental, et. al., 1991;Baker and
Halpin,1987).The discrepancy of the results in different experiments might be
449

450
M. Sahraei et al.
relate to the difference in the age of the chicks when feeding high Zn commenced,
duration of feeding, dietary Zn concentration, and previous body stores.
Conclusion
The results of this study showed that in corn soybean meal diets the
highest RBV of different zinc sources were observed in 150ppm concentration,
and Bioplex Zn was more bio-availabile than zinc sulfate or zinc oxide in
practical diets.
Procena relativne biološke dostupnosti nekoliko izvora cinka
u ishrani brojlera konvencionalnom obrokom
M. Sahraei, H. Janmmohamadi, A. Taghizadeh, G. Ali Moghadam, S. A. Rafat
Rezime
Ogled je izveden sa brojlerskim pilićima muškog pola, u fazi porasta (8-
28d) u cilju procene biološke dostupnosti četiri izvora cinka (Zn), cink sulfat
(ZnSO4.H2O), dva izvora cink oksida (ZnO FG1 i ZnO FG2) i Bioplex Zn. Cink
sulfat (ZnSO4.H2O) je korišćen kao standard u ispitivanju biološke dostupnosti.
Pilići su nasumično podeljeni u 13 prehrambenih tretmana sa po 6 grla po
ponavljanju i 4 ponavljanja po tretmanu, koji je uključivao obrok na bazi kukuruza
i sojine sačme (25.50 mg Zn / kg SМ), ili osnovni obrok dopunjen sa 100, 150 ili
200 mg / kg SМ bilo kao ZnSO4.H2O (33 % Zn), cink oksid FG1(72% Zn), cink
oksid FG2(75% Zn) ili Bioplex Zn(15% Zn). Nivo Zn u obroku kao i izvor nisu
imali uticaj (P>0.05) na unos hrane ili prirast brojlera tokom prve i druge nedelje
eksperimentalnog perioda, ali kod parametara konverzija hrane u prvoj i drugoj
nedelji ogleda, kao i unosa hrane, prirasta i konverzije u trećoj nedelji, kao i u
celom ogledu, utvrđene su signifikante razlike između tretmana (P

References
Estimation of the relative bioavailability ...
AMMERMAN C. B.(1995): Methods for estimation of mineral bioavailability.
Pages 83–92, in Bioavailability of Nutrients for Animals.C. B. Ammerman, D. H.
Baker, and A. J. Lewis, ed. AcademicPress, San Diego, CA.
ANONYMOUS.(1982):Analytical Methods for Atomic Absorption
Spectrophotometry, Perkin-Elmer Corp., Nor Walk, CT.
AOAC.(1995):Official Methods of Analysis. 16th ed. Association of Official
Analytical Chemists, Arlington, VA.
AOYAGI S. and D.H. BAKER. (1993):Nutritional evaluation of copper-lysine and
zinc lysine complexes for chicks. Poultry Sci. 72:165-171.
BAKER, D.H. and K.M. HALPIN.(1987): Efficacy of a manganese-protein chelate
compared with that of manganese sulfate for chicks. Poultry Sci. 66:1561-1563
BURRELL, A. L., W. A. DOZIER, A. J. DAVIS, M. M. COMPTON, M.
E.FREEMAN, P. F. VENDERLL, and T. L. WARD.(2004): Response of broilers
to dietary zinc concentrations and sources in relation to environmental
implications. Br. Poult. Sci. 45:255–263.
CAO, J., X. G. LUO, P. R. HENRY, C. B. AMMERMAN, R. C. LITTEL, and R.
D. MILES(1996): Effect of dietary iron concentration, age, and length of iron
feeding on feed intake and tissue iron concentration of broiler chicks for use as a
bioassay of supplemental iron sources. Poult. Sci. 75:495–504.
CAO, J., P. R. HENRY, R. GUO, R. A. HOLWERDA, J. P. TOTH, R. C. LITTEL,
R. D. MILES, and C. B. AMMERMAN.(2000):Chemical characteristics and
relative bioavailability of supplemental organic zinc sources for poultry and
ruminants. J. Anim. Sci.78:2039–2054.
CAO, J., P. R. HENRY, S.R, DAVIS, R.J. COUSINS, R. D. MILES, R.C.
LITTELL and C. B. AMMERMAN.(2002): Relative bioavailability of organic zinc
sources based on tissue zinc and metallothionein in chicks fed conventional dietary
zinc concentrations. Anim. Feed Sci. Technol. 101:161-170.
CHENG, J., E. T. KORNEGY and T. SCHELL.(1998): Influence of dietary lysine
on the utilization of zinc from zinc sulfate and a zinc lysine complex by young
pigs. J. Anim. Sci. 76:1064-1074.
COLLINSE, N. E., and E. T. MORAN, Jr.(1999): Influence of supplemental
manganese and zinc on live performance and carcass quality of broilers. J. Appl.
Poult. Res.8:222–227.
DUNCAN, D. B.(1955): Multiple range and multiple F tests. Biometrics 11:1-42.
EDWARDS, Jr., H. M. (1959): The availability of chicks to zinc in various
compounds and ores. J. Nutr. 69:306-308.
EDWARDS, H. M., III, S. D. BOLING, J. L. EMMERT, and D. H.
BAKER.(1998): Bioavailability of zinc in two zinc sulfate by-products of the
galvanizing industry. Poultry Sci. 77:1546−1549.
451

452
M. Sahraei et al.
FALCHUK, K. H. and B. L. VALLEE. (1985): Zinc and chromatin structure,
composition and function. In: Trace elements in man and animals (Ed. C. F. Mills,
I. Bremner and J. K. Chesters).CAB Publishing, UK, pp. 48-55.
FINNEY, D. J.(1978): Statistical Method in Biological Assay (3rd Ed.).Charles
Griffin and Co., London.
EMMERT, J. L., and D. H. BAKER.(1995): Zinc stores in chickens delay the onset
of zinc deficiency symptoms.Poult. Sci. 74:1011–1021.
KAIM, W. and B. SCHWEDERSKI.(1994):Bioinorganic chemistry: inorganic
elements in the chemistry of life. John Wiley and Sons Ltd, England. pp. 401.
LEDUXE, D.R.(2005): Bioavailability and antagonists of trace minerals in
ruminant metabolism. Porc. 2005 Florida Ruminant Nutrition Sym., 16 th Annual
Meeting, Gainesville, Florida, pp.23-37.
LEESON S .(2003): A new look at the trace mineral nutrition of poultry: Can we
reduced environmental burden of poultry manure? In Nutritional Biotechnology in
the Feed and Food Industries. T.P. Lyons and K.A. Jacques,ed. Nottingham Univ.
Press, Nottingham, UK.
LI, S., X. LUO, B. LIU, T. D. CRENSHAW, X. KUANG, and G.
SHAO.(2004):Use of chemical characteristics to predict relative bioavailability of
supplemental organic manganese sources for broilers. J.Anim. Sci. 82:2352–2363.
LI, S. F., X. G. LUO, L. LU, T. D. CRENSHAW, Y. Q. BU, B. LIU, X.KUANG,
G. Z. SGAO, and S. X. YU. (2005): Bioavailability of organic manganese sources
in broilers fed high dietary calcium.Anim. Feed Sci. Technol. 123:703–715.
LITTELL, R. C., P. R. HENRY, A. J. LEWIS, and C. B. AMMERMAN. (1997):
Estimate of relative bioavailability of nutrients using SAS procedures.J. Anim. Sci.
75:2672–2683.
Luo, X. G., S. F. Li, L. Lu, B. Liu, X. Kuang, G. Z. Shao, and S.
X.Yu.(2007):Gene expression of manganese-containing superoxide dismutase as a
biomarker of manganese bioavailability for manganese sources in broilers. Poult.
Sci. 86:888–894.
MILLER, E. R., P. K. KU, J. P. HITCHCOCK, and W. T. MAGEE. (1981):
Availability of zinc from metallic zinc dust for young swine. J.Anim. Sci. 52:312–
315.
NRC.(1994): Nutrient Requirements of Poultry.9th rev. ed. Natl. Acad. Press,
Washington, DC.
NOLLET, L. G. HUYGHEBAERT, and P. SPRING.(2008): Effect of different
levels of dietary organic(Bioplex) trace minerals on live performance of broiler
chickens by growth phases. J. Appl. Poult. Res. 17:109–115.
doi:10.3382/japr.2007-00049
PIMENTAL, J.L., M.E. COOK, and J.L. GREGER.( 1991): Bioavailability of zinc
methionine
for chicks. Poultry Sci. 70:1637-1639.

Estimation of the relative bioavailability ...
REVY, P.S., JONDREVILLE, C., DOURMAD, J.Y. and NYS, Y.(2003): Le zinc
dans l’alimentation du porc:oligo-élément essential et risqué potentiel pour
l’environnement. INRA Production Animale 16 : 3-18.
ROBERSON, R. H., and P. J. SCHAIBLE. (1958): The zinc requirement of the
chick. Poult. Sci. 37:1321–1323.
ROBERSON, R. H., and P. J. SCHAIBLE. (1960): The availability to the chicks of
zinc as the sulfate, oxide or carbonate. Poult. Sci.39:835-837.
SAS Institute. (1999): SAS Statistics User’s Guide. Statistical Analytical System.
5th rev. ed. Carry, NC, SAS Institute Inc.
SANDOVAL, M., P. R. HENRY, C. B. AMMERMAN, R. D. MILES and R.C.
LITTELL.(1997): Relative bioavailability of supplemental inorganic zinc sources
for chicks. J. Anim. Sci. 75:3195-3205.
STAHL, J. L., M. E. COOK, and M. L. SUNDE. (1986): Zinc supplementation: Its
effect on egg production, feed conversion, fertility and hatchability. Poult. Sci.
65:2104–2109.
SPEARS, J. W.(1989): Zinc methionine for ruminants: Relative bioavailability of
zinc in lambs and effects of growth and performance of growing heifers. J. Anim.
Sci. 67:835-843.
WEDEKIND, K. J. and D. H. BAKER.(1990): Zinc bioavailability in feed-grade
sources of zinc. J. Anim. Sci. 68:684-689.
WEDEKIND, K. J. A. E. HORTIN and D. H. BAKER .(1992): Methodology for
assessing zinc bioavailability: Efficacy estimates for zinc-methionine, zinc sulfate,
and zinc oxide. J.
Anim. Sci. 70:178-187.
WEDEKIND, K. J., A. J. LEWIS, M. A. GIESEMANN, and P. S.
MILLER.(1994): Bioavailability of zinc from inorganic and organic sources for
pigs fed corn-soybean meal diets. J. Anim. Sci. 72:2681–2689.
WEIGAND, E. and KIRCHGESSNER, M.(1980):Total true efficiency of zinc
utilization determination and homeostatic dependence upon to zinc supply status in
young rats. Journal of Nutrition 110: 469-480.
ZEIGLER, T. R., R. M. LEACH, L. Jr, C. NORRIS, and M. L. SCOTT.
(1961):Zinc requirement of the chick: Factors affecting requirement. Poult. Sci.
40:1584–1593.
Received 8 march 2012; accepted for publication 15 May 2012
453

Biotechnology in Animal Husbandry 28 (3), p 455-462 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.082’4
DOI: 10.2298/BAH1203455V
HERITABILITY AND REPEATABILITY ESTIMATES OF
REPRODUCTION TRAITS IN PUREBRED PIGS
V. Vidovic 1 , D. Lukac 1 , M. Stupar 1 , V. Višnjic 2 , J. Krnjaic 3
1
Faculty of Agriculture, Department of Animal Science, Trg Dositeja Obradovica 8, 21 000 Novi Sad,
Republic of Serbia
2
SIZIM DOO, Veliki Otok bb, 48 317 Legrad, Croatia
3
Delta Agrar, Napredak AD, Golubinacki put bb, 22300 Stara Pazova, Republic of Serbia
Corresponding author: vidovic.vitomir@gmail.com
Original scientific paper
Abstract: Research was performed on four farms, on 2434 highly fertile
females Landrace and Yorkshire, and 28 boars of Danish origin, or 7684
consecutive parities, in period 2009 - 2012. Study of genetic parameters of
conventional breeds Landrace and Yorkshire were conducted on 3964 females who
mated with 49 males or 15764 litters in the same period. Estimates of genetic
parameters for litter size show the same tendency as the legality of the purebred
sows that produce 11-14 weaned piglets less per sow per year. Environmental
factors, HYS, food technology and management expressed no significant effect on
traits. Heritability and repeatability of live and still born piglets, litter size and the
live at day 5 after birth and the number of piglets weaned in category of low
hereditary traits whose values vary within the limits of 0.07 to 0.12 for the
heritability and from 0.15 to 0.19 for the repeatability. There was tendency of
lower values of genetic parameters in the conventional compared to highly fertile
sows, which is considered to be the effect of selection on gene frequency for the
observed traits.
Key words: genetic parameters, reproduction traits, pigs
Introduction
The genetic parameters are used by producers and breeders, primarily to
study the natural action of genes, namely additive ones. Also that can be used for
modelling of selection criteria, and selection direction of breeding. The nucleus of
the farms is genetically superior material, completely healthy and used to produce
desirable effects in the selection of pure breed and hybrid animals of both sexes.
The current strategy of selection and crossbreeding showed significant
improvement when talking about litter size at birth and weaning. This is obtained
by selecting Landrace and Yorkshire pigs, particularly those originating from
Denmark, which produce 11-14 more weaned piglets per sow per year over the

456
V. Vidovic et al.
existing selection. The selection in our conditions showed almost identical pattern.
A similar trend was also observed in hybrid sows. We analysed the value of
population genetic parameters in sows that were highly fertile sows and the same
breed, national - conventional selection, which produce significantly less piglets
per unit of time.
The purpose of study is to estimate and compare values of genetic
parameters for traits like: litter size and weight at birth and weaning in female with
such a high level of fertility compared to the conventional local selection of the
same breed.
Materials and methods
Research was performed on 2434 female Landrace and Yorkshire animals,
and 28 boars of Danish origin, born locally, or 7684 consecutive parities, in the
period 2009 - 2012. Study of the genetic parameters of conventional Landrace and
Yorkshire breeds were conducted on 3964 females who mated with 49 boars and
produced 15764 litters in the same period.
Mixed model equations (MME) was used to analyse available data, the model 1.
Yijkl = µ + Ki + GSij + Pijk * Eijkl
Yijkl – the observed traits;
µ - general mean value;
Ki – random sire effect;
GSij – fixed effect of differences between farms, years and seasons;
Pijk – effect of farrowing (parity);
Eijkl – random error.
Analysed traits are: age of sows at first farrowing, number of live and still born
pigs, live born piglets five days after birth, litter size and weight at weaning.
Duration of lactation was 28 days. Diet of lactating sows was ad libitum and
determined within the efficient technology, according to stage of lactation and
production. Technology diet was the same at all stages of production.
Results and Discussion
The results are shown in Tables 1-9. Age of sows at first farrowing was
366 in Landrace and 372 days in Yorkshire breed. Sows conventional Landrace
and Yorkshire were younger 32 and 42 days compared to highly fertile animals.
Indirectly, conventional sows were lighter before the insemination and farrowing
for 29-32 kg body weight (Table 1).

Heritability and repeatability ...
Table 1. Age at 1 st farrowing, body weight at insemination and farrowing of prolificacy and
conventional selection of Landrace and Yorkshire
Traits
Prolificacy
Landrace Yorkshire
Conventional
Landrace Yorkshire
Age at 1 st
farrowing
366 372 334 330
Weight at
138 142 109 110
insemination
Weight at
farrowing
188 195 156 163
Litter size at farrowing and weaning did not differ significantly in some
farrowings between the two breeds, but within the breed with the expected trend,
and therefore have identical criteria for selection. There were no differences in
weight or the number of piglets at weaning. Number of piglets five days after birth
was less than the number of live born to 0.7. These differences are small pigs,
which are the result of high selection pressure by increasing the genotype of this
traits, i.e., more piglets at farrowing, and the possibility of optimal nutrition in
piglets to be uniform on born. Also, this is partly a consequence of the lack of milk
for such a large number of piglets born. These two traits are important and open
questions for future work of breeders and nutritionists (Table 2 and 3). Almost the
same is the case in conventional breeds. However, the highly fertile breed, in
average, has higher values of genetic parameters for all analysed properties which
is considered the contribution of selection.
Table 2. Litter size of prolificacy and conventional selection of Landrace
Farrowing Prolificacy Conventional
Alive born Still born Weaned Alive born Still born Weaned
1. 13.8 2.3 13.1 10.8 0.9 9.3
2. 14.3 2.4 13.6 11.3 1.3 10.6
3. 15.0 2.8 13.8 11.6 1.3 10.8
4. 14.4 2.7 13.6 11.9 1.7 11.0
5. 14.1 2.6 13.4 12.1 1.9 11.1
1+2+3+4+5 14.4 2.5 13.6 11.4 1.6 10.9
Table 3. Litter size of prolificacy and conventional selection of Yorkshire
Farrowing
Prolificacy Conventional
Alive born Still born Weaned Alive born Still born Weaned
1. 13.6 2.4 12.9 10.0 0.9 9.0
2. 14.3 2.4 13.7 10.2 1.3 10.0
3. 14.9 2.6 13.9 11.1 1.6 10.3
4. 14.0 2.3 13.5 11.3 1.8 10.6
5. 14.8 2.8 13.8 11.1 2.4 10.4
1+2+3+4+5 14.5 2.5 13.5 10.6 1.9 10.2
457

458
V. Vidovic et al.
All the factors (sire, FYS and farrowing) have a significant influence on
observed traits. Similar results have been reported by (Vidovic et al., 2011a; Vincek
2005; Radojkovic et al., 2005: Vidovic, 1976). This indicator shows that despite the
modern conditions of keeping pigs in a farm where automatic control of production
parameters such as light, humidity and temperature, are not balanced with females
genome, which means that research is needed to improve the environment.
Farrowing had a significant impact on the observed traits of both conventional as
and highly fertile sows meaning that further studies are needed to stabilize the
genome of these breeds.
Table 4. The heritability of litter size of prolificacy Landrace
Farrowing Age at
farrowing Alive born Still born 5 th day Weaned
1. 0.15 0.12 0.05 0.10 0.11
2. - 0.11 0.02 0.09 0.10
3. - 0.09 0.05 0.08 0.10
4. - 0.09 0.03 0.08 0.09
5. - 0.09 0.04 0.10 0.08
1+2+3+4+5 - 0.10 0.03 0.09 0.10
Heritability and repeatability were not significantly different compared to
conventional selection of breeds which have much lower genetic potential (Vidovic
et al., 2011b; Vidovic et al., 2011c; Tretinjak et al., 2009; Chen et al., 2003; Lucia
et al., 2002; Crump et al., 1997; Gordon 1997; Hue et al., 1993, Petrovic et
al.,1991). This justifies the use of new knowledge in the area optimization of
environment, optimization technology feeding sows and age with weight the
introducing of the reproduction.
Table 5. The heritability of litter size at conventional selection of Landrace
Farrowing Age at
Born
farrowing Alive born Still born 5 th day Weaned
1. 0.12 0.10 0.05 0.11 0.10
2. - 0.09 0.02 0.09 0.10
3. - 0.09 0.02 0.09 0.09
4. - 0.08 0.04 0.11 0.11
5. - 0.07 0.05 0.10 0.11
1+2+3+4+5 - 0.10 0.04 0.11 0.11

Heritability and repeatability ...
Table 6. The heritability of litter size of prolificacy Yorkshire
Farrowing Age at
Born
farrowing Alive born Still born 5 th day Weaned
1. 0.21 0.12 0.03 0.07 0.10
2. - 0.09 0.02 0.08 0.10
3. - 0.10 0.04 0.09 0.12
4. - 0.09 0.03 0.09 0.08
5. - 0.09 0.02 0.07 0.11
1+2+3+4+5 - 0.11 0.03 0.08 0.11
Table 7. The heritability of litter size of conventional selection of Yorkshire
Farrowing Age at
Born
farrowing A Live born Still born 5 th day Weaned
1. 0.14 0.12 0.04 0.10 0.10
2. - 0.08 0.05 0.08 0.08
3. - 0.09 0.02 0.09 0.09
4. - 0.09 0.03 0.07 0.07
5. - 0.11 0.03 0.07 0.09
1+2+3+4+5 - 0.10 0.03 0.08 0.09
Table 8. The repeatability of litter size of prolificacy Landrace
Farrowing Age at
farrowing Alive born Still born 5 th day Weaned
1. 0.20 0.16 0.09 0.17 0.18
2. - 0.16 0.08 0.15 0.18
3. - 0.18 0.09 0.16 0.17
4. - 0.18 0.07 0.18 0.17
5. - 0.18 0.08 0.17 0.16
1+2+3+4+5 - 0.17 0.09 0.17 0.17
Table 9. The repeatability of litter size of conventional selection of Landrace
Farrowing Age at
Born
farrowing Alive born Still born 5 th day Weaned
1. 0.19 0.18 0.07 0.16 0.16
2. - 0.16 0.08 0.16 0.15
3. - 0.17 0.08 0.17 0.16
4. - 0.17 0.07 0.15 0.18
5. - 0.18 0.07 0.15 0.20
1+2+3+4+5 - 0.18 0.07 0.16 0.19
459

460
Conclusion
V. Vidovic et al.
There are statistically significant differences for number of alive, still born
and weaned piglets between prolificacy and conventional Landrace and Yorkshire
breeds. No differences within prolificacy are for the same traits. Prolificacy’s L and
Y gilts were heavier at the entrance to the production, which guarantees the
continuity of a stable production of pigs and optimal replacement. Conventional
selection of the same L and Y breeds were, by selection criteria and technology, at
least one month younger and with smaller size. These indicate shorter life
production and smaller litter size. Genetic parameters indicate that reproduction
belong to low hereditary traits regardless of the level of genetic potential and
significantly larger litters than the conventional selection of these breeds in our
conditions. Even statistically significant differences on phenotypic level, the
estimation of genetic parameters for both indicated they are belonging to low
heritage group of traits. Knowing that ovulation rate belong to middle heritage trait
(h 2 – 0.40) it is necessarily to use different management in feeding regime of
females and age with weight of gilts. Research justifies the use of new information
technologies in the area of feeding sows, and optimization of production
conditions, e.g. humidity, air velocity, temperature and light, which significantly
affect the development of the genome. A particularly important aspect is the
adaptation of modern management aspects in production management at the farm.
As expected evaluations the repeatability values were significantly higher than
heritability. The differences between genetic parameters of prolificacy and
conventional purebred L and Y belong to selection efficiency during generation.
Heritabilnost i ponovljivost reprodukcijskih svojstava svinja
čistih rasa
V. Vidović, D. Lukač, M. Stupar, V. Višnjić, J. Krnjaić
Rezime
Istraživanja su izvedena na 4 farme na kojima su krmače podeljene u dve
grupe: visokoplodne i konvencionalne sa znatno nižom plodnošću. Broj
visokoplodnih iznosio je 2.434 krmača rase landras i jorkšir koje su parene sa 28
nerastova,svi Danskog porekla ili, 7.684 legla iz prvih 5 prašenja a u periodu 2009.
– 2012. Uporedo su izvedena i istraživanja na plotkinjama koje su odabirane
koristeći konvencionalne selekcsijske kriterijume, rasa landras i jorkšir, koja su
obuhvatila 3.964 krmače parene sa 49 nerastova, i koje su proizvele 15.764 legla u
istom periodu. Na obe selekcije, visokoplodne i konvencionalne, primenjeni su

Heritability and repeatability ...
različiti kriterijumi selekcije (danski i srpski) tokom godina proizvodnje sa
očekivanim efektima. Za korekciju uticaja sistematskih i slučajnih faktora korišćen
je mešoviti model ocene. Razlika u broju zalučene prasadi između visokoplodnih i
konvencionalnih selekcija bila je signifikantna, a varirala je u granicama 11-14
zalučene prasadi na godišnjem nivou. Ocene heritabilnosti varirale su u granicama
0.07 – 0.12 dok su vrednosti koefcijent ponovljivosi bile veće i to za nivo dodatnih
informacija iz modela. Varirale su u granicama 15 – 19 procenata. Selekcijski
kriterijumi u konvencionalnih selekcija nisu bili efikasni prema očekivanju. Bili su
pristrasni. Visokoplodne životinje pokazale su genetsku superiornost ali je udeo
aditivnih gena bio gotovo identičan kao u konvencionalni selekcija obe rase.
Reference
CHEN P., BASS T. J.,MABRY J. W., KOEHLER K. J., DEKKERS C. M. (2003):
Genetic Parameters and Trends for Litter in U.S. Yorkshire, Duroc, Hampshire,
and Landrace Pigs. J. Anim Sci., 81, 46-53.
CRUMP R. E., HALEY C. S., THOMSON R., MERCER J. (1997): Individual
Animal Model Estimates of Genetic Parameters for Reproduction Traits of
Landrace Pigs Performance Tested in a Commercial Nucleus Herd. J. Animal Sci.,
65, 285-290.
GORDON I. (1997): Controlled Reproduction in Pigs. Oxon, CAB International,
pp. 247.
HUE L. L., DIAL G. D., MARSH W. E., DAVIES P. R., MOMONT H. W.
(1993):Influence of Lactation Length on Sow Productivity. Livest. Prod. Sci.,34,
253-265.
LUCIA T. J., CORREA N. M., DESCHAMPS C. J., BIANCHI I., DONIN A. M.,
MACHADO C. A., MEINCKE W., MATHEUS E. M. J. (2002): Risk Factors for
Stillbirths in Two Swine Farms in the South of Brazil. Preventive Veterinary
Medicine, 53, 285-292.
PETROVIC M., LATINOVIC D., STOJIC P., KOSOVAC O. (1991): Uticaj
genetskih i faktora okoline na varijabilnost rezultata performans testa nazimica.
Biotehnologija u stočarstvu, 7, (1-2): 25-32.
RADOJKOVIC D., PETROVIC M., MIJATOVIC M., RADOVIC I. (2005):
Phenotypic and Genetic Correlation of Fertility Traits of Swedish Landrace.
Biotechnology in Animal Husbandry, 21 (3-4): 79-88.
TRETINJAK M., SKORPUT D., DJIKIC M., LUKOVIC L. (2009): Veličina legla u
krmača na obiteljskim gospodarstvima u republici Hrvatskoj. Stočarstvo, 63, 175-185.
VIDOVIC V., VIŠNJIC V., JUGOVIC D., PUNOS D., VUKOVIC N. (2011a):
Praktično svinjarstvo. APROSIM. Novi Sad, pp. 287.
VIDOVIC V., LUKAC D. (2010): Genetika životinja, Poljoprivredni fakultet Novi
Sad, pp 361.
461

462
V. Vidovic et al.
VIDOVIC V. (1997): Poređenje ocene genetskih parametara za reprodukcijske osobine
svinja gajenih u čistoj rasi i ukrštanju. Doktorska disetacija, Biotehnički fakultet, Ljubljana.
VIDOVIC V., LUKAC D., STRBAC LJ., STUPAR M. (2011b): Effect of age and
weight of Yorkshire gilts at mating on litter size and longevity. Stočarstvo, 65, 3-12.
VIDOVIC V., STRBAC LJ., LUKAC D., STUPAR M. (2011c): Influence of age
and weight of Landrace gilts at fertile insemination on litter size and longevity.
Biotechnology in Animal Husbandry, 27, 75-85.
VINCEK D.(2005): Veličina legla majčinskih linija uzgojnog programa u
svinjogojstvu. Stočarstvo, 59, 13-21.
Received 16 may 2012; accepted for publication 15 August 2012

Biotechnology in Animal Husbandry 28 (3), p 463-468 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.082’4
DOI: 10.2298/BAH1203463R
THE EFFECT OF GENOTYPE AND YEAR ON TRAITS
OF PERFORMANCE TESTED GILTS
Č. Radović 1 , M. Petrović 2 , N. Parunović 3 , N. Brkić 4 , B. Živković 1 , M.
Gogić 1 , N. Stanišić 1
1
Institute for Animal Husbandry, Autoput 16, 11080 Belgrade-Zemun, Republic of Serbia
2
Faculty of Agriculture University of Belgrade, Nemanjina 6, 11080 Belgrade, Republic of Serbia
3
Institute of Meat Hygiene and Technology, Kaćanskog 13, 11000, Belgrade, Republic of Serbia
4
Ministry of Agriculture, Forestry and Water Management - Directorate for Water, 11000, Belgrade,
Republic of Serbia
Corresponding author: cedomirradovic.izs@gmail.com
Original scientific paper
Abstract: Objective of this research was to establish the effect of animal
genotype, year of measuring and birth on traits of performance tested gilts: age at
the end of the test (AET), life daily gain (LDG), back fat thickness 1 and 2 (BFT1
and BFT2), depth of back muscle (MLD) and assessed share of meat in carcass. Of
total number of animals (n=3600) included in the research, 1709 animals were of
genotype SL and 1891 of genotype ♀SLx♂LY. In regard to the birth year of tested
gilts, the distribution was following: n2006=296, n2007=895, n2008=934, n2009=803,
n2010=589 and n2011=83 gilts. In regard to testing year, the distribution of gilts was
as follows: n2007=682, n2008=875, n2009=962, n2010=697 and n2011=384 gilts. The
effects of test year and birth year were established (P0.05).
Key words: gilts, genotype, ultra sonogram measurements, PigLog 105
Introduction
Important precondition in the work on genetic improvement of the quality
of pigs is knowledge of the variability of production traits of breeding animals. For
that purpose, studies are carried out and data recorded for all traits included in the
selection procedure. Efficiency in pig production is evaluated based on three
parameters: annual sow productivity, feed conversion ratio and meat yield of
animals. Quantative and qualitative carcass traits depend on selection methods. It is
known that certain quantitative pig traits are not equaly passed on, which means
that the possibilities of their improvement through selection are different.
Heritability coefficients for growth traits and traits of carcass side quality are
medium to strong (Hermesch et al., 2000; Chen et al., 2002; Gorjanc et al., 2003;

464
Č. Radović et al.
Radović et al., 2003; Petrović et al., 2006). Increased selection for meat content
causes significant decrease of the content of subcutaneous fat (Bahelka et al.,
2007). Breeding value of young animals can be used for evaluation of parent
breeding value, and in that way the best parent combinations selected for breeding
and eventually significantly higher production and better quality of pig meat can be
ensured in a country (Jukna et al., 2009). In the study of the average values of
production traits of sons and daughters, Petrović et al., (2002) have established
expressed effect of sires (P

The effect of genotype and ...
the values according to test years, i.e. to the end of test, we can see that animals
from the first to the fifth year prolonged the test from year to year (GM1:GM5;
198:212 days), and hence the life daily gain decreased (GM1:GM5; 500:465 g).
During the same test period also decrease of the back fat thickness (BFT1 and
BFT2) by 6.9 mm and 5.3 mm, respectively, was observed. In the same testing
period the decrease in back fat thickness was recorded. By observing the data
according to birth year (GR1:GR6) of gilts there is a same trend of decrease of
values for back fat thickness BFT1 and BFT2. For said traits the effect of year of
measuring and year of birth (P0.05).
Table 1. The effect of genotype and year on intensity of growth and back fat thickness
(LSM±S.E.)
Sources of variation AET 3) , days LDG, g BFT1, mm BFT2, mm
μ ± S.E. 203.56±0.56 486.21±1.43 17.66±0.41 13.45±0.41
Genotype
Year of
measuring
Birth year
1 1)
2
204.46±1.92
202.65±1.91
483.71±4.74
488.72±4.73
18.01±0.76
17.31±0.76
13.46±0.67
13.43±0.67
P 2) NS NS NS NS
1
2
3
4
5
197.85±0.95
199.67±0.81
203.96±0.75
204.19±0.81
212.10±1.03
500.09±2.36
496.08±2.01
485.04±1.87
484.71±2.03
465.15±2.55
21.37±0.49
19.65±0.45
17.17±0.44
15.65±0.45
14.47±0.51
16.28±0.46
15.28±0.44
12.87±0.43
11.84±0.44
10.97±0.49
P *** *** *** ***
1
2
3
4
5
6
198.86±1.49
201.95±0.89
198.57±0.72
199.56±0.71
213.09±0.82
209.31±1.08
496.82±3.68
489.82±2.22
498.93±1.80
496.69±1.79
463.10±2.05
471.94±2.69
20.91±0.63
19.33±0.47
17.98±0.43
17.31±0.43
16.40±0.46
14.04±0.52
16.40±0.57
15.01±0.45
13.39±0.43
13.09±0.43
11.85±0.44
10.93±0.49
P *** *** *** ***
1) 1-SL, 2- LYxSL; 2) NS=P>0,05; *=P

466
Č. Radović et al.
Table 3. The effect of genotype and year on MLD depth and share of meat (LSM±S.E.)
Sources of variation MLD 3) , mm Share of meat, %
μ ± S.E. 52.79±0.22 55.04±0.39
Genotype
1 1)
2
53.06±0.76
52.52±0.76
54.89±0.66
55.18±0.66
P 2) NS NS
1
51.40±0.38
51.76±0.45
2
51.87±0.32
53.09±0.43
Year of measuring 3
53.08±0.30
55.60±0.42
4
53.60±0.32
56.86±0.43
5
54.00±0.41
57.87±0.47
P *** ***
1
51.49±0.43
52.57±0.56
2
52.38±0.33
53.50±0.44
Birth year
3
4
52.91±0.28
53.05±0.29
54.87±0.41
55.32±0.41
5
53.17±0.35
56.30±0.43
6
53.74±0.59
57.67±0.48
P ** ***
1) 2)
1-SL, 2- LYxSL; NS=P>0,05; *=P

The effect of genotype and ...
animals. By observing the values according to year of measuring, i.e. to the end of
test, we can conclude that animals in the period through first five years of
measuring have prolonged the test from year to year, which influenced the decrease
of life daily gain. In the same period of measuring, the decrease in back fat
thickness (BFT1 and BFT2) by 6.9 mm and 5.3 mm was observed. By observing
the values according to year of birth (GR1:GR6) of gilts, the same trend of
decrease of values for back fat thickness BFT1 and BFT2 is observed. For all
studied traits the effect of year of measuring and year of birth (P0.05).
Acknowledgements
Research was financed by the Ministry of Education and Science of the
Republic of Serbia, project TR31081.
Uticaj genotipa i godine na osobine performans testiranih
nazimica
Č. Radović, M. Petrović, N. Parunović, N. Brkić, B. Živković, M. Gogić, N.
Stanišić
Rezime
Cilj ovog istraživanja je da se utvrdi uticaj genotipa grla, godine merenja i
rođenja grla na osobine performans testiranih nazimica: uzrast na kraju testa
(UKT), životni dnevni prirast (ŽDP), debljina slanine 1 i 2 (DSL1 i DSL2), dubina
leđnog mišića (MLD) i procenjeni udeo mesa u trupu. Od ukupnog broja (n=3600)
istraživanjem je obuhvaćeno 1709 grla genotipa ŠL i 1891 grla genotipa
♀ŠLx♂VJ. Po godini rođenja testiranih nazimica distribucija je bila sledeća:
n2006=296, n2007=895, n2008=934, n2009=803, n2010=589 i n2011=83 nazimice. Dok je
po godinama testiranja distribucija nazimica bila sledeća: n2007=682, n2008=875,
n2009=962, n2010=697 i n2011=384 nazimice. Za sve ispitivane osobine utvrđen je
uticaj godine merenja i godine rođenja (P0.05).
References
BAHELKA I., TOMKA J., HANUSOVA E. (2007): The effects of probe type and
intensity of ultrasound on accuracy of intramuscular fat prediction in Longissimus
dorsi muscle of pigs. Biotechnology in Animal Husbandry, 23, 5-6, 87-95.
467

468
Č. Radović et al.
CHAN D. E., WALKER N. P., MILLS W. E. (2002): Prediction of Pork Quality
Characteristics Using Visible and Near-Infrared Spectroscopy. Transactions of the
ASAE, 45, 5, 1519-1527.
GOGIĆ M., M. PETROVIĆ, B. ŽIVKOVIĆ, Č. RADOVIĆ, D. RADOJKOVIĆ,
N. PARUNOVIĆ, G. MARINKOV (2012): Uticaj različitih faktora na osobine
performans testiranih nazimica. Biotechnology in Animal Husbandry, 28, 2, 312-321.
GORJANC G., MALOVRH Š., KOVAČ M., GLAVAČ-VNUK M., ZRIM J.
(2003): Proučavanje možnosti vključivte klavnih lastnosti v napoved plemenske
vrednosti pri prašičih. Zbornik Biotehniške Fakultete Univerze v Ljubljani,
Kmetijstvo Zootehnika, 82, 2, 89-96.
HARVEY R.W. (1990): User's guide for LSMLMW and MIXMDL. Ver. PC–2, 1–91.
HERMESCH S., LUXFORD B.G., GRASER H.U. (2000): Genetic parameters for
lean meat yield, meat quality, reproduction and feed effciency traits for Australian
pigs. 1. Description of traits and heritability estimates. Livestock Production
Science, 65, 239–248.
JUKNA V., JUKNA Č., PEČIULAITIENE N., MEŠKINUTE-KAUŠILIENE E.
(2009): The effect of boars and sows on meat quality and calorific values in the
progeny. Biotechnology in Animal Husbandry, 25, 3-4, 161-172.
MICHALSKA G., NOWACHOWICZ J., BUCEK T., WASILEWSKI D. P.
(2008): Changes in range of performance test results of gilts of Polish Large White
breed produced in Poland in Bydgoszcz breeding region. Journal Central European
Agriculture, 9, 3, 581-588.
NOWACHOWICZ J., MICHALSKA G., BUCEK T., WASILEWSKI P. D.
(2009): Meat and fat content of crossbred gilts born and kept in Poland in
Bydgoszcz breeding district in years 1995-2004. Journal Central European
Agriculture, 10, 4, 367-374.
PETROVIĆ M., D. RADOJKOVIĆ, D. ROMIĆ, M. PUŠIĆ, M. MIJATOVIĆ, N.
BRKIĆ (2002): Genetska i fenotipska varijabilnost osobina performans testiranih
nerastova i nazimica. Biotechnology in Animal Husbandry 18, 5-6, 67-72.
PETROVIĆ M., M. PUŠIĆ, D. RADOJKOVIĆ, M. MIJATOVIĆ,Č. RADOVIĆ,
B. ŽIVKOVIĆ (2006): Fenotipska i genetska varijabilnost osobina kvaliteta polutki
i mesa. Biotehnologija u stočarstvu, 22, 5-6, 1-10.
RADOVIĆ Č., PETROVIĆ M., JOSIPOVIĆ S., ŽIVKOVIĆ B., KOSOVAC O.,
FABJAN M. (2003): Uticaj različitih genotipova, očeva i sezone klanja na klanične
osobine svinja. Biotehnologija u stočarstvu, 19, 1-2, 11-16.
SZYNDLER-NEDZA M., TYRA M., RÓZYCKI M. (2010). Coefficients of
heritability for fattening and slaughter traits included in a modified performance
testing method. Ann. Anim. Sci., 10: 117–125.
Received 8 march 2012; accepted for publication 15 May 2012

Biotechnology in Animal Husbandry 28 (3), p 469-475 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.08’4
DOI: 10.2298/BAH1203469P
HERITABILITY AND CONNECTIONS OF SOW
FERTILITY TRAITS
M. Popovac 1 , D. Radojković 1 , M. Petrović 1 , M. Mijatović 1 , M. Gogić 2 ,
D. Stanojević 1 , N. Stanišić 2
1
Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade-Zemun, Republic of
Serbia
2
Institute for Animal Husbandry, Autoput 16, 11080, Belgrade-Zemun, Republic of Serbia
Corresponding author: mlp@agrif.bg.ac.rs
Original scientific paper
Abstract: Purpose of this paper was to determine fertility traits heritability
coefficients of the sows (number of live born, total number of born, stillborn and
reared piglets in the litter) and interconnections between these traits. Heritability
coefficients were low and averaged in interval from h 2 = 0,056 for number of
reared piglets in litter to h 2 = 0,142 for total number of born piglets in litter, which
is in accordance with heritability values for reproductive traits. Genetic
interconnections of these traits had wide variation interval and averaged from r = -
0,221 between number of still born and reared piglets in litter to r = 0,947 between
total number of born and number of live born piglets in litter. Coefficients of
phenotype correlation varied in interval from r = -0,162 between number of still
born and number of live born piglets in litter to r = 0,909 between total number of
born and number of live born piglets in litter.
Key words: sow, heritability, genetic and phenotype connection
Introduction
Parental genetic influence at daughter fertility is relatively low if we take
that reproductive traits are highly conditioned by the environment factors.
However, even beside that if we were to make genetic improvement it is needed to
choose those parents that have best results in reproduction. As reproductive boars
have greater number of offspring it is needed to have strict selection, greater
number of offspring (daughters) furthermore gives opportunity to get more
accurate results at progeny testing of same boars.
Greater number of research that refer to size of the litter in different
population of pigs showed that heritability coefficients and repetitiveness are low
(10-15%), and that variation coefficient were relatively high (25%).

470
M. Popovac et al.
Most frequently evaluated genetic trait of pigs is the number of live born
piglets in the litter. According to Bereskin (1984) value of the heritability
coefficient for this trait is 0,05 while Skorupski et al., (1996) determined the value
of 0,19. Next to the number of live born piglets in litter, very frequently traits like
total number of born piglets, number of stillborn piglets and number of reared
piglets in liter were researched, which was done in this research too.
After determining the heritability coefficient for fertility traits it was
necessary to determine correlation between researched traits, which is shown by
phenotype and genetic correlation coefficients. Absolute values and sign in front of
these coefficients point how much and how these traits are interconnected.
Therefore knowing about values of these coefficients gives the possibility to alter
certain traits in such manner that values of other traits are not affected.
Value for phenotype correlation coefficient which was determined by
Popov et al., (2003) between number of live born and still born piglets in the litter
was 0,218. Relatively low value of this coefficient shows weak correlation between
these two traits, much larger coefficient was determined for genetic correlation
coefficient 0,803 between these two traits. The high value of genetic correlation
coefficient was also determined by Roehe and Kennedy (1995). This shows that
with value increase of one trait the value of other trait increases too, and vice versa.
Material and methods
A criterion on which calculation of heritability and correlation coefficient
was based was that fathers of analyzed sows have 10 and more daughters in the
researched group of sows. Based on this criterion 54 boars and their 1318
daughters which had 4489 litters were taken. Fertility traits for which heritability
and their phenotype and genetic correlation was calculated were: total number of
born piglets in litter (TNBPL), number of live born piglets in litter (NLBPL),
number of still born piglets in litter (NSBPL) and number of reared piglets in litter
(NRPL).
Starting analysis has rounded descriptive statistic data rendering where
arithmetic mean and indicators of variation (variation coefficient, standard
deviation and variation interval) were calculated. Mathematic-statistical data
processing was done by applying the mix model, method of least squares and
application of program package by Harvey (1990) within which using the method
of interclass correlation of half-sisters heritability was calculated as well as genetic
and phenotype coefficients. In the variance analysis model based on which
heritability for specific traits was calculated, beside the sow father following
factors were included: farm, genotype, furrowing year, furrowing season,
furrowing order, number of teats and age at first furrowing. For these factors it was
determined that they have significant statistic influence on researched traits,

Heritability and connections ...
however those factors will not be discussed further. In this way more accurate
heritability coefficient was determined and in certain measure surrounding
variability influence was brought to minimum. Presented results were calculated
with help of following mix model:
Yijklmnop = µ + Ri + Fj + Gk + Sl + Pm + Bn + Uo + op+ eijklmnop
Yijklmno – value of observed trait (TNBPL, NLBPL, NSBPL, NRPL),
µ - population average,
Ri – fixed genotype influence,
Fj – fixed farm influence,
Gk – fixed furrowing year influence,
Sl – fixed furrowing season influence,
Pm – fixed furrowing order influence,
Bn – fixed influence of teat number,
Uo – regression influence of age at first furrowing,
op – random father influence,
eijklmnop – influence of undetermined factors (error).
Research results and discussion
Heritability of researched traits in the Table 1. Values of heritability coefficients
for traits: total number of born (TNBPL), live-born (NLBPL), stillborn (NSBPL)
and number of reared (NRPL) piglets in the litter of researched sows.
Table 1. Heritability values determined by analysis
Trait Heritability and heritability standard error
TNBPL 0,142 ± 0,034
NLBPL 0,123 ± 0,031
NSBPL 0,083 ± 0,024
NRPL 0,056 ± 0,020
Results show that values of heritability are low and that they vary in the
interval from 0,056 to 0,142. Highest heritability value h 2 =0,142 is noted for total
number of born piglets in litter. Notably more important trait like number of live
born piglets has slightly lower value of h 2 =0,123, which represents strong influence
of non genetic factors on this trait. Heritability coefficient for NLBPL in this
research is somewhere in between the varying interval for this coefficient (0,05 –
0,19) which was determined by Skorupski et al., (1996) and Bereskin (1984) in
their research.
Opposite to these traits number of reared piglets in litter has the lowest
value of h 2 =0,056 because of the strong influence of the surrounding at the number
of reared piglets (handling with piglets, diet, care and housing of sow and piglets).
471

472
M. Popovac et al.
Roehe and Kennedy (1995) have determined almost the same value for this trait in
their research.
When we look at the number of stillborn piglets in the litter of researched
sows it has been determined that, that value is influenced with h 2 =0,083 by genetic
factors. Of course it would have been ideal if number of stillborn piglets were to be
equal to zero, however as that is not possible the goal is to keep that number as low
as possible. As it is quoted by Popov et al., (2003) heritability for number of still
born piglets is 0,017 which is almost five times lower value compared to value that
was determined in this analysis. Furthermore according to Roehe and Kennedy
(1995) heritability value is 0,06, while Southwood and Kennedy (1990) quote that
heritability for this trait is 0,13. Determined value for NSBPL heritability is within
limits of values that were determined by the last two groups of researchers. All this
is pointing that number of stillborn piglets is significantly more influenced by other
factors than it is influenced by heritability.
Correlation between the researched fertility traits. In order to see the
correlation and in order to interpret it in a best way genetic and phenotype
correlation coefficients will serve as best example. Correlation coefficients
between researched traits show inter-correlation between the same. Correlation
coefficients for fertility traits of researched sows are shown in Table 2.
Table 2. Genetic correlation coefficients for fertility traits and their significance 1)
Trait NLBPL NSBPL NRPL
TNBPL 0,947 **
0,429 **
0,619 **
NLBPL - 0,117 **
0,759 **
NSBPL - - -0,221 **
1)
Correlation coefficient for 5 and 1% of certainty (d.f.=1000) is 0,062 and 0,081.
High value of genetic correlation coefficient (r=0,947, total correlation)
between total number of born and number of live born piglets is understandable
and it shows that by selection for higher number of live born piglets in litter there is
increase of total number of born piglets. High value for genetic correlation
coefficient 0,947 between total number of born and number of live born piglets in
litter was also determined by Roehe and Kennedy (1995).
Genetic correlation coefficient between NLBPL and NSBPL was 0,117
which shows low genetic correlation compared to correlation (0,803) that was
determined by Popov et al., (2003).
In the Table 2. negative value for genetic correlation coefficients is also
shown, which points that during selection for one trait, value of other trait
decreases and vice versa. Even though a very low value -0,221 (very low
correlation), between number of reared and number of still born piglets, it shows
that generally in researched litters with higher number of reared piglets was less
stillborn piglets.

Heritability and connections ...
However, low absolute values for genetic correlation coefficient give the
possibility for improvement of one trait with selection without significantly
violating the value of the other trait.
Phenotype correlation coefficients show more objective correlation
influence because it is not possible to clearly separate surrounding environment
and genetic factors influence on trait. Phenotype correlation coefficients besides
genetic factors include the environment factors.
Table 3. Phenotype correlation coefficients for fertility traits 1)
Trait NLBPL NSBPL NRPL
TNBPL 0,909 **
0,264 **
0,227 **
NLBPL - -0,162 **
0,281 **
NSBPL - - -0,133 **
1) Correlation coefficient for 5 and 1% certainty (d.f. =1000) is 0, 062 and 0,081.
In the Table 3. high value for phenotype correlation coefficient is shown
(r=0,909, total correlation) between total number of born piglets and number of live
born piglets, as it was for genetic correlation which shows that in production
environment with value increase for one trait, the value of other trait is also
increased. Result is almost identical to the result determined by Radojković et al.
,(2005).
Number of reared and totally born piglets in litter in this research has low
coefficient of phenotype correlation (r = 0,227, very low correlation). Gajić and
Radivojević (1980) have also in their research determined lower value for this
coefficient 0,265, while Lui et al., (1982) have determined high correlation for this
trait and the coefficient was 0,930.
Table 3. also shows negative values for correlation coefficients which point
that with increase of value for one trait, the value of other trait is decreasing.
Negative value for phenotype correlation coefficient -0,162 (very low correlation)
was determined between number of live born piglets and number of stillborn
piglets in litter. Results are not in agreement with results determined by Popov et
al., (2003) who have determined the positive value (0,218) of phenotype
correlation coefficient for these two traits.
It has to be emphasized that all correlation coefficients are statistically
significant. Level and strength of correlation is determined based on values that
were given in the tables by Latinović (1996).
Conclusion
Heritability coefficients were in the interval between 0,056 for NRPL and
up to 0,142 for TNBPL. These low values of heritability are connected to
reproductive traits and they show how much surrounding environment factors
significantly influence the fertility of the pigs. Moreover, these low values of
473

474
M. Popovac et al.
heritability for fertility traits emphasize the need for bettering the surrounding
environment condition which has strong influence on researched traits.
All genetic and phenotype correlation coefficients were statistically
significant. The lowest genetic correlation (0,117) was between NLBPL and
NSBPL, while almost total genetic correlation (0,947) was determined between
NLBPL and TNBPL.
Determined heritability and correlation coefficient values show that there is
possibility for selection and improvement of significant reproductive traits, which
should result in genetic progress for researched pig population.
Acknowledgements
Research was financed by the Ministry of Education Science, and Technological
Development, Republic of Serbia, project TR 31081.
Naslednost i povezanost osobina plodnosti krmača
M. Popovac, D. Radojković, M. Petrović, M. Mijatović, M. Gogić, D. Stanojević,
N. Stanišić
Rezime
Cilj rada bio je da se utvrde koeficijenti naslednosti osobina plodnosti
krmača (broj živorođene, ukupno rođene, mrtvorođene i odgajene prasadi u leglu) i
međusobna povezanost ovih osobina. Koeficijenti heritabiliteta su bili niski i
kretali su se u intervalu od h 2 = 0,056 za broj odgajene prasadi u leglu do h 2 = 0,142
za broj ukupno rođene prasadi u leglu, što je u skladu sa vrednostima heritabiliteta
za reproduktivne osobine. Genetska povezanost ovih osobina imala je širok
interval variranja i kretala se od r = -0,221 između broja mrtvorođene i broja
odgajene prasadi u leglu, do r = 0,947 između broja ukupno rođene i broja
živorođene prasadi u leglu. Koeficijenti fenotipske korelacije kretali su se u
intervalu od r = -0,162 između broja mrtvorođene i broja živorođene prasadi u
leglu, do r = 0,909 između broja ukupno rođene i broja živorođene prasadi u leglu.
References
BERESKIN B. (1984): A genetic analysis of sow productivity traits. Journal of
Animal Science, 59, 5, 1149-1163.
GAJIĆ I., RADIVOJEVIĆ R. (1980): Značaj i veličina genetskih promena iz
direktne i indirektne selekcije svinja. Savremena poljoprivreda, 28, 3-4, 111-121.

Heritability and connections ...
HARVEY R.W. (1990): User's guide for LSMLMW and MIXMDL. Ver. PC–2, 1–91.
LATINOVIĆ D. (1996): Populaciona genetika i oplemenjivanje domaćih životinja.
Praktikum, Beograd, 1-173.
LUI J.F., GIANNONI M.A., BANZATTO D.A., CARREGAL R.D. (1982): Razao
sexual, taxa mortalidade, numero medio de tetas e correlacoes lineares entre
caracteristicas de leitegades das racas Duroc E Landrace. Revista da Sociedade
Brasileira de Zootehnica, 11, 1, 1-13.
POPOV R., RADOVIĆ I., TRIFUNOVIĆ S., TEODOROVIĆ M., PETROVIĆ M.
(2003): Fenotipska ispoljenost, varijabilnost i naslednost osobina plodnosti svinjabroj
mrtvorođene prasadi u leglu. Savremena poljoprivreda, 52, 3-4, 297-302.
RADOJKOVIĆ D., PETROVIĆ M., MIJATOVIĆ M., RADOVIĆ I. (2005):
Fenotipska i genetska povezanost osobina plodnosti plotkinja švedskog landrasa.
Biotechnology in Animal Husbandry, 21, 3-4, 79-88.
ROEHE R., KENNEDY B. W. (1995): Estimation of genetic parametars for litter
size in Canadian Yorkshire and Landrace swine with each party of farrowing
treated as a different trait. Journal of Animal Science, 73, 10, 2959-70.
SKORUPSKI M. T., GARRICK D. J., BLAIR H. T. (1996): Estimates of direct
and maternal genetic parameters for production and reproduction traits in tree
breeds of pigs. New Zeland Journal of Agriculture Research, 39, 3, 387-395.
SOUTHWOOD O. I., KENNEDY B. W. (1990): Estimation of direct and maternal
genetic variance for litter size in Canadian Yorkshire and Landrace swine using an
animal model. Journal of Animal Science, 68, 7, 1841-1847.
Received 4. April 2011; accepted for publication 16. May 2012
475

Biotechnology in Animal Husbandry 28 (3), p 477-486 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 637.04
DOI: 10.2298/BAH1203477O
MEAT FATTY ACID PROFILE OF PIGS FED LINSEED
ENRICHED DIET
Đ. Okanović, D. Ivanov, D. Palić, A. Mandić, N. Ilić
University of Novi Sad, Institute of Food Technology, Bulevar Cara Lazara 1
21000 Novi Sad, Serbia
Corresponding author: djordje.okanovic@fins.uns.ac.rs
Original scientific paper
Abstract: The aim of this study was to evaluate the influence of diet
supplemented with linseed rich additive on fatty acid profile and omega-fatty acids
content in pig meat. Twelve pigs were divided in a control and experimental group
and grown to 110 kg of live weight. The experimental group was fed a standard
diet enriched with 2.5% of commercial additive Vitalan ® . After the end of feeding
period, the meat samples from both groups were analyzed for fatty acids content in
raw and roasted meat. The ratio between omega-6 and omega-3 acids was
established. Samples from experimental group fed with linseed enriched diet
showed higher ω-3 acids content in meat (6.24% - 7.23%), compared to the control
group (0.71% - 1.64%), thus making it better for a human consumption from a
health perspective. Linseed enriched diet positively influenced fatty acid
composition of pig M. Longissimus dorsi muscle by decreasing SFA content, as
well as increasing PUFA and UFA content. Heat treatment did not significantly
decreased content of stearic and linoleic fatty acids in any sample. It was concluded
that the diet enriched with extruded linseed had beneficial effect on the majority of
monitored parameters in the study.
Key words: n -3 polyunsaturated fatty acids, linseed, pig meat
Introduction
Many clinical, epidemiological and biological studies suggest that dietary
fats play an important role in human health and well being (Gebauer et al., 2006).
Results from those studies ascribe a particular significance to n-3 polyunsaturated
fatty acids (PUFA). This class of fatty acids are found to have anti-inflammatory,
antithrombogenic, and hypotriglyceridemic properties, they are also active against
some types of cancer like colon, breast and prostate cancer (Connor, 2000). n-3
PUFA consumption also reduces risk of cardiovascular disease. At the same time
increased levels of n-6 fatty acids are associated with an increase in chronic
diseases (Givens et al., 2006).

478
Đ. Okanović et al.
In paper, Sretenović et al., (2009), the significance of omega-3 fatty acids
in human nutrition is presented. As essential substances they cannot be synthesized
in the organism, but have to be introduced through diet. Also, the significance of
some essential omega-6 fatty acids as well as their mutual relation, are presented.
The role of omega-3 fatty acids in animal nutrition is also pointed out in this paper,
introduced or consumed by animals either by grazing or as diet supplement, which
influence improvement of their production, reproduction and health performances.
The main reason for this is production of animal feed from grains rich in n-6 fatty
acids which subsequently leads to meat rich in the same type of fatty acids
(Crawford, 1968). Being aware of n-3 PUFA benefits and health promoting effects,
the nutritionists recommend a diet rich in n-3 fatty acids as well as a lower n-6/n-3
ratio from the current 15-20:1 to 1-4:1 (Simopoulos, 2002). Since animals are not
able to synthesize n-3 and n-6 fatty acids and animal diet determines the fatty acid
composition in meat, change in fatty acids ratio and composition in meat can be
achieved through the change in animal diet (Mourot and Hermier, 2001; Ivanov et
al., 2009). Most common way to perform this is by enriching the animal feed with
fish oil/fish meals or with plant oils rich in n-3 PUFA or seed meals such as linseed
(Raes et al., 2004; Kouba et al., 2003; Stevanović et al., 2011). Monogastric
animals (like pigs) are better target for this approach because the dietary fatty acids
are absorbed from the intestine unchanged. In this way, composition of fatty acids
in meat could be modified by dietary means ultimately improving nutritional and
health value of the meat (Enser et al., 2000).
The objective of this study was to investigate the influence of linseed
enriched diet (rich in n-3 PUFA) on fatty acid composition in raw and roasted pig
meat. The overall aim of the study was to improve the nutritional value of pig meat
using linseed enriched diet, thus making it a good source of beneficial n-3 fatty
acids.
Materials and Methods
Animals and diet
Total twelve pigs, Pietrain x (Landrace x Great Yorkshire) were used in the
study, which has been conducted at the pig farm „Sabo Janos”, Jermenovci, Serbia.
Twelve pigs were divided into two groups and fed with two types of diet, a
standard diet and diet enriched with 2.5% of Vitalan ® (Vitalac, France), until
reaching approximate 110 kg of live weight, when they were slaughtered. Vitalan ®
contains 85% extruded linseed, which made the diet rich in omega-3 acids and the
rest were wheat bran and antioxidants. The composition of the diets is shown in
Table 1.

Table 1. Composition of diets for pigs
Meat fatty acid profile of ...
Components (%) Control group Experimental group
Vitalan® 2.5
Maize 51.0 50.0
Barley 28.0 26.8
Soybean meal 18.0 17.7
Premix 2.5 2.5
Acidifier 0.5 0.5
Total 100.0 100.0
Slaughtering and sampling
The animals were slaughtered and samples of pig meat (M. Longissimus
dorsi, bacon and back fat), 200 g each, from both groups were collected and kept in
the refrigerator at 4 0 C until further use. A half of the samples were roasted in the
oven at the temperature of 80 - 85 0 C until the temperature in the centre of the meat
reached 69 0 C (about 1 hour).
After 24 hours, the samples were sent to the laboratories of Food
Technology Institute in Novi Sad, where fatty acid analysis and sensory evaluation
were performed.
Fat extraction for fatty acid analysis
Supercritical fluid extraction with CO2 was used for preparation of fat
extracts, as recommended for fatty acid analysis (Ivanov et al., 2011). Extractions
were performed on a LECO TFA2000 fat analyzer with methods developed in the
laboratory. Temperature, pressure and extraction flow rates were adopted from
existing LECO procedures (LECO Corporation, 2003). Cell temperature and flow
path temperatures were set at 100 °C. The collection vials on the instrument are not
temperature-controlled and remained near room temperature, approximately 30 °C.
Operating pressure was 9000 psi. Extraction flow rate was set at 1.3 l/min.
Infusorial soil (LECO Corporation, 2003) was used to disperse the sample and
when used, modifier was added to the mix prior to transferring the sample to the
extraction thimble. From the extracted lipids, fatty acid methyl esters were
prepared with method that use boron trifluoride/methanol solution (Veresbaranji,
1996). Nitrogen gas was used for drying and removing solvents from fatty acid
methyl esters. Obtained samples were analyzed by a GC Agilent 7890A system
with FID, auto-injection module for liquid and headspace sampling, equipped with
fused silica capillary column (DB-WAX 30 m, 0.25 mm, 0.50 µm). Helium was
used as a carrier gas (purity > 99.9997 vol. %, flow rate = 1.26 ml/min). The fatty
acids peaks were identified by comparison of retention times with retention times
of standards from Supelco 37 component fatty acid methyl ester mix and with data
479

480
Đ. Okanović et al.
from internal data library, based on previous experiments. Results were expressed
as mass of fatty acid or fatty acid group (g) per 100 g of fatty acids, and as a ratio
between omega-6 and omega-3 fatty acids.
Statistical analysis
STATISTICA software version 9 (Statsoft, Tulsa, OK, USA) was used for
analyzing variations (analysis of variance – ANOVA) and for Tukey’s HSD
comparison of means of samples (Statistica, 2006).
Results and Discussion
Fatty acid composition of the meat (M. longissimus dorsi), heat treated
meat, fat and bacon of pigs fed with standard and linseed enriched diet is shown in
Table 2.
The major fatty acids in all tissues (listed from most prevalent to least)
were oleic (C18:1), linoleic and other isomers of C18:2, palmitic (C16:0),
palmitoleic (C16:1) and stearic (C18:0). These five fatty acids accounted for over
90% of the total fatty acids in the pig tissues.
The data showed that fatty acid compositions of the examined samples
were broadly similar. However, there were important species differences. As it is
known from the literature, pigs have relatively high proportion of linoleic acid
(C18:2 n-6), major PUFA, which is entirely derived from the diet (Wood et al.,
2008). The column used for gas chromatographic determination did not have
possibility to separate positional isomers, in combination with used standard
mixture, but it could be noticed that all experimental samples (in pigs fed the
experimental diet) have had higher content of total C18:2 fatty acid, to use of
linseed enriched diet.
From the results shown in Table 2 for fresh and roasted pig meat, bacon
and back fat, it is evident that use of linseed enriched diet resulted in increased
levels of omega-3 fatty acids in the experimental group (6.24% - 7.23%) compared
to the control group (0.71% - 1.64%). This significantly contributed to the more
favourable ratio of omega-6/omega-3 in the experimental group (3.35-3.91)
compared to the control group (11.96-31.49).
Results for the content of free fat in both examined groups have shown that
the proportion of free fat in the control and experimental groups were similar. Meat
is the major source of fat including SFA in the human diet. SFA have been
generally labeled as the cause of cancers and coronary heart disease.

Meat fatty acid profile of ...
Table 2: Fatty acid composition (%) of different pig tissues
Meat Roasted meat Back fat Bacon
Sample C E C E C E C E
Free fat (%) 5.11 5.57 6.02 5.34 86.50 82.90 28.10 26.10
Fatty acid Fatty acid content (% of total fatty acid)
C10:0 0.08 0.05 0.09 0.07 0.06 0.06 0.09 0.07
SD 0.02 0.06 0.07 0.05 0.03 0.05 0.06 0.03
C12:0 0.04 0.03 0.04 0.04 0.04 0.04 0.04 0.04
SD 0.01 0.02 0.02 0.03 0.03 0.02 0.01 0.03
C14:0 0.75 0.57 0.75 0.63 0.72 0.72 0.77 0.67
SD 0.24 0.26 0.34 0.09 0.36 0.29 0.31 0.19
C16:0 8.98 6.90 9.09 7.31 9.00 8.99 8.89 7.79
SD 0.89 0.71 1.21 0.96 1.01 0.24 0.35 0.41
C16:1 9.25 8.08 9.47 8.99 7.08 6.04 9.01 8.77
SD 0.67 0.58 0.91 0.87 0.61 0.89 1.03 0.99
C17:0 0.47 0.71 0.41 0.72 0.64 0.64 0.43 0.71
SD 0.23 0.09 0.16 0.65 0.13 0.21 0.13 0.51
C17:1 0.42 0.66 0.36 0.69 0.42 0.42 0.41 0.64
SD 0.16 0.51 0.24 0.29 0.31 0.14 0.52 0.16
C18:0 6.83* 4.94* 7.33* 4.85* 7.33 7.13 6.62 5.31
SD 0.87 0.68 0.19 0.34 0.16 0.65 0.94 0.36
C18:1 c+t 48.91** 42.49** 50.61* 44.38* 48.15 48.71 50.59** 41.11**
SD 1.06 0.98 1.03 1.01 0.89 0.88 0.97 0.79
C18:2 19.61** 24.22** 19.96** 24.53** 22.36** 27.20** 20.07** 26.11**
SD 0.97 0.89 0.76 1.02 0.96 0.87 0.96 1.01
C18:3n3 1.64** 7.23** 0.69** 6.24** 0.71** 7.20** 0.79** 6.67**
SD 0.62 0.89 0.46 0.79 0.51 0.96 0.49 0.69
C20:0 0.13 0.10 0.12 ND 0.16 0.14 0.13 0.10
SD 0.06 0.09 0.11 - 0.14 0.08 0.05 0.09
C20:1 1.05 0.93 1.04* 0.47* 1.45 1.44 1.13* 0.70*
SD 0.25 0.21 0.34 0.12 0.64 0.61 0.56 0.23
C20:2 0.65* 1.01* 0.60 0.77 0.88 0.88 0.73 0.79
SD 0.22 0.52 0.14 0.31 0.19 0.17 0.11 0.25
C20:4 0.33 0.32 0.30 ND ND ND ND ND
SD 0.15 0.14 0.10 - - - - -
C20:3 n3+n6 0.85 3.66 ND 0.97 ND ND ND ND
SD 0.21 0.41 - 0.12 - - - -
C18:2 / C18:3n3 11.96 3.35 28.93 3.93 31.49 3.78 25.41 3.91
SFA 17.28 13.30 17.83 13.61 17.95 17.73 16.98 14.69
MUFA 59.63 52.17 61.49 54.53 57.10 56.61 61.14 51.23
PUFA 23.09 36.44 20.86 32.52 23.95 35.28 21.59 33.57
UFA 82.72 88.60 82.35 87.05 81.05 91.89 82.73 84.79
MUFA/SFA 3.45 3.92 3.45 4.01 3.18 3.19 3.60 3.49
PUFA/SFA 1.34 2.74 1.17 2.39 1.33 1.99 1.27 2.29
- control; E – experimental; SD – standard deviation ND – not determined; SFA – saturated fatty
acid; MUFA – monounsaturated fatty acid; PUFA – polyunsaturated fatty acid; UFA – total
unsaturated fatty acid,
*-statistically significant difference between the experimental and control samples of the same group
within the row, at p < 0.05
**-statistically significant difference between the experimental and control samples of the same group
within the row, at p < 0.01
481

482
Đ. Okanović et al.
The average ratio of PUFA/SFA recommended by the British Department
of Health is more than 0.45, and WHO/FAO experts have reported guidelines for a
“balanced diet” in which suggested ratio of PUFA/SFA should be above 0.4 (Wood
et al., 2003; HMSO. 1994). In our study, that ratio was significantly higher (p <
0.05) than recommended values in all samples (from 1.17 to 2.74). Scollan et al.
(2006), reported that high values of PUFA/SFA ratios in pig M longissimus dorsi
are consequence of high level of linoleic acid (C18:2 n-6). In our study, after heat
treatment of M. longissimus dorsi muscle, there was no significant (p > 0.05)
decrease of PUFA/SFA ratio, but between results in pig muscles fed with linseed
enriched diet, there was a significant increase (p = 0.02) of PUFA/SFA ratio, in
comparison with control samples.
Δ5 and Δ6 desaturase and elongase enzymes have an important role to
synthesize long chain (C20-22) PUFA from 18:2n-6 and 18:3n-3. Muscles
contained very high proportion of these long chain fatty acids, unlike fat and
bacon, where C20:4 and C20:3 n3+n6 were not even detected. Although these fatty
acids show positive nutritional effect, recommendation is that their ratio in tissue
should not be more than 4.0 (Wood et al., 2003; Scollan et al., 2006), which have
been the case in all our samples, except for the samples E (Table 2).
Comparison of fatty acid content in M. longissimus dorsi muscle before
and after heat treatment showed that there was no significant change (p > 0.05) in
fatty acid composition between these samples (Fig. 1).
On the other hand, samples of M. longissimus dorsi of the control and
experimental groups showed significant differences in fatty acid composition (Fig.
2). SFA content decreased significantly (p = 0.006), MUFA content also
significantly (p = 0.04) decreased, and PUFA and UFA contents significantly (p =
0.03 and p = 0.02, respectively) increased in experimental groups.
SFA – saturated fatty acid, MUFA – monounsaturated fatty acid,
PUFA – polyunsaturated fatty acid; UFA – total unsaturated fatty acid
Figure 1. Changes of fatty acid composition after heat treatment in M. longissimus dorsi muscle
of pigs fed with control and linseed enriched diet

Meat fatty acid profile of ...
SFA – saturated fatty acid, MUFA – monounsaturated fatty acid; PUFA – polyunsaturated fatty acid,
UFA – total unsaturated fatty acid; Levels of significance: *p < 0.05 for results of the same group;
**p < 0.01 for results of the same group
Figure 2. Fatty acid composition in M. Longissimus dorsi muscle of pigs fed with control and
linseed enriched diet
Levels of significance: *p < 0.05 for results of the same group; **p < 0.01 for results of the same
group
Figure 3. Content of the stearic and linoleic acid in M. longissimus dorsi of pigs fed control and
linseed enriched diet
It is known that UFA containing double bonds are easily oxidized, and thus
fatty acid composition can influence the palatability of meat, as well as that the
effect of fatty acids on meat tenderness is due to the different melting points of
individual fatty acids, especially stearic and linoleic acid (Jeong et al., 2010).
483

484
Đ. Okanović et al.
In Figure 3 are shown changes in the content of these two acids and their
total amounts in M. longissimus dorsi, as a result of the differences in composition
of standard (control) and linseed enriched diet used for feeding of pigs.
As it can be seen from the Fig. 3, linseed enriched diet significantly
influenced the content of both stearic and linoleic acid (p = 0.013 and p = 0.003,
respectively). Although the stearic acid content decreased, the content of linoleic
acid increased to a level that the sum of these two acids was significantly higher in
comparison with the meat of pigs fed the control diet. After the heat treatment, the
content of those fatty acids were lower, in both, control and experimental samples,
but the differences were insignificant (p > 0.05). Linseed enriched diet positively
influenced the content of stearic and linoleic acid, and therefore it might positively
affected the tenderness of M. longissimus dorsi muscle. Heat treatment did not
significantly decrease the content of these fatty acids in any sample, and
consequently, probably do not showed significant interference with the tenderness
of pig meat (Okanović et al., 2010).
Conclusions
Based on the results of this study, the following conclusions can be made:
- Levels of omega-3 fatty acids in the fresh and roasted meat of M. longissimus
dorsi, bacon and back fat in experimental group were much higher than in the
control group, which made the ratio of omega-6 to omega-3 fatty acids much
more favourable toward n-3 fatty acids in the experimental group.
- From nutritional point of wiev, our study justified the use of the extruded
linseed enriched diet to improve the ratio of omega-6 and omega-3 fatty acids to
the desired level less than 4.
- Linseed enriched diet positively influenced the fatty acid composition of pig M.
longissimus dorsi muscle by decreasing SFA content, while increasing PUFA
and UFA contents.
- Heat treatment did not significantly decreased content of stearic and linoleic
fatty acids in any sample.
Our studies could contribute to improvement of the nutritional value of pig meat by
use of linseed enriched diet, thus making it a good source of beneficial n-3 fatty
acids.
Acknowledgement
The experimental work represented in this paper is a part of Integrated and
Interdisciplinary Research Project funded by Serbian Ministry of Education,
Science and Technological Development, Project No: III 46012.
Masnokiselinski profil mesa svinja hranjenih smešom
obogaćenom semenom lana
Đ. Okanović, D. Ivanov, D. Palić, A. Mandić, N. Ilić

Rezime
Meat fatty acid profile of ...
Cilj istraživanja bio je da se utvrdi uticaj smeše sa dodatkom aditiva
bogatog lanenim semenom na masnokiselinski profil i sadržaj omega-masnih
kiselina u svinjskom mesu.
Dvanaest grla podeljeno je u dve grupe: kontrolnu i eksperimentalnu, i
uzgajano do težine od 110 kilograma. Eksperimentalna grupa je hranjena
standardnom smešom obogaćenom sa 2.5% komercijalnog aditiva Vitalan ® . Na
kraju perioda uzgoja analiziran je sadržaj masnih kiselina svežeg i pečenog mesa
uzoraka uzetih iz obe grupe. Takođe je utvrđen odnos između omega-6 i omega-3
masnih kiselina.
Uzorci iz eksperimentalne grupe hranjene smešom obogaćenom lanom
pokazali su viši sadržaj omega-3 masnih kiselina u mesu, čineći ih poželjnijim sa
stanovišta ljudske ishrane i zdravlja. Ishrana obogaćena lanom pozitivno je uticala
na sastav masnih kiselina u mišiću M. Longissimus dorsi snižavajući sadržaj
zasićenih masnih kiselina i povećavajući sadržaj mono- i polinezasićenih masnih
kiselina. Toplotni tetman nije značajno uticao na sadržaj stearinske i linolne
kiseline ni u jednom uzorku. Na kraju je zaključeno da ishrana obogaćena
ekstrudiranim lanenim semenom ima blagotvoran uticaj na većinu parametara koji
su praćeni tokom eksperimenta.
References
CONNOR W.E. (2000): Importance of n-fatty acids in health and disease.
American Journal of Clinical Nutrition, 71, 171S-175S.
CRAWFORD M.A. (1968): Fatty acid ratios in free-living and domestic animals.
Lancet I, 1329-1333.
ENSER, M., RICHARDSON, R.I., WOOD, J.D., GILL, B.P. & SHEARD, P.R.
(2000): Feeding linseed to increase the the n-3 PUFA of pork: fatty acid
composition of muscle, adipose tissue, liver and sausages. Meat Science, 55, 201-212.
GEBAUER S.K., PSOTA T.L., HARRIS W.S., KRIS-ETHERTON P.M. (2006):
n-3 Fatty acid dietary recommendations and food sources to achieve essentiality
and cardiovascular benefits. American Journal of Clinical Nutrition .83(suppl).
1526S-1535S
GIVENS, D.I., KLIEM, K.E., GIBBS, R.A. (2006): The role of meat as a source of
n-3 polyunsaturated fatty acids in the human diet. Meat Science, 74, 209-218.
HMSO. UK DEPARTMENT OF HEALTH (1994): Nutritional aspects of
cardiovascular disease. London. Report on Health and Social subject. 46. 37 – 46.
IVANOV, D., ČOLOVIĆ, R., BERA, O., LEVIĆ, J., SREDANOVIĆ, S., (2011):
Supercritical fluid extraction as a method for fat content determination and
preparative technique for fatty acid analysis in mesh feed for pigs. Eur. Food Res.
Technol. 233 (2), 343-350
IVANOV, D., LEVIĆ, J., SREDANOVIĆ, S., (2009): Fatty acid composition of
various soybean products, Food and Feed Research, 38 (2), 65-70.
485

486
Đ. Okanović et al.
JEONG, D.W., CHOI, Y.M., LEE, S.H., CHOE, J.H., HONG, K.C., PARK, H.C.,
KIM, B.C. (2010): Correlations of trained panel sensory values of cooked pork
with fatty acid composition, muscle fiber, and pork quality characteristics in
Berkshire pigs. Meat Science, 86, 607 – 615.
KOUBA, M., ENSER, M., WHITTINGTON, F.M., NUTE, G.R. & WOOD, J.D.
(2003): Effect of a high-linolenic acid diet on lipogenic enzyme activities, fatty
acid composition and meat quality in the growing pig. Journal of Animal Science,
81, 1967-1979.
LECO Corporation (2003): Organic application note, St. Joseph, MI
http://www.leco.com/resources/application_notes/pdf/TFE2000_MEATS_CRUDE
_FAT_AOAC_PVM_3-2000_203-821-122.pdf
MOUROT, J. & HERMIER, D. (2001): Lipids in monogastric animals meat.
Reproduction Nutrition Development, 41, 109-118.
OKANOVIĆ, Đ., PALIĆ, D., ILIĆ, N., 2010. Influence of addition of linseed on
the diet on meat quality of piglets. Quality of Life 1 (2-4), 134-138
RAES, K., DE SMET, S. & DEMEYER, D. (2004): Effect of dietary fatty acids on
incorporation of long chain polyunsaturated fatty acids and conjugated linoleic acid
in lamb, beef and pork meat: a review. Animal Feed Science and Technology, 113,
199-221.
SCOLLAN, N., HOCQUETTE, J.F., NUERNBERG, K., DANNENBERG, D.,
RICHARDSON, I. & MOLONEY, I. (2006): Innovations in beef production
systems that enchance the nutritional and health value of beef lipids and their
relationship with meat quality. Meat Science, 74, 17 – 33.
SIMOPOULOS A.P. (2002): The importance of the ratio of omega-6/omega-3
essential fatty acids. Biomedicine and Pharmacotherapy, 56, 365-379.
SRETENOVIĆ, L., PANTELIĆ, V., NOVAKOVIĆ, Ž., (2009): Importance of
utilization of omega-3 fatty acids in human and animal nutrition. Biotechnology in
Animal Husbandry, 25(5-6-1), pp. 439-449.
STATISTICA (Data Analysis Software System), v.8.0. Stat-Soft, Inc, USA (www.
statsoft.com) (2006):
STEVANOVIĆ, J., PALIĆ, D., OKANOVIĆ, Đ. (2011): Legislation and risks of
use of nutritional feed supplements. Food and Feed Research, 38(1), 21-26.
VERESBARANJI, I. (1996): In Karlović, Đ. & Andrić, N. (Eds.) Kontrola
kvaliteta semena uljarica (pp. 311-314). Tehnološki fakultet Novi Sad, Savezni
zavod za standardizaciju, Beograd.
WOOD. J.D.. RICHARDSON. R.I.. NUTE. G.R.. FISHER. A.V.. CAMPO. M.M..
KASAPIDOU. E. SHEARD. P.R. & ENSER. M. (2003): Effect of fatty acids on
meat quality: A review. Meat Science. 66. 21 – 32.
WOOD, J.D., ENSER, M., FISHER, A.V., NUTE, G.R., SHEARD, P.R,
RICHARDSON, R.I., HUGES, S.I. & WHITTINGTON, F. M. (2008): Fat
deposition, fatty acid composition and meat quality: A review. Meat Science, 78,
343 – 358.
Received 17 May 2012; accepted for publication 5 July 2012

Biotechnology in Animal Husbandry 28 (3), p 487-496 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636
DOI: 10.2298/BAH1203487D
THE EFFECT OF THE LEVEL OF MILK YIELD ON THE
REPRODUCTION TRAITS IN BLACK AND WHITE
COWS
R. Djedović 1 , V. Bogdanović 1 , G. Trifunović 1 , M. D. Petrović 2 , M. M.
Petrović 3 , D. Stanojević 1
1
Department of ZOO Techniques, University of Belgrade, Faculty of Agriculture, Nemanjina 6,
Zemun, Republic of Serbia
2
Faculty of Agronomy, Cara Dušana 34, 32000, Čačak, Republic of Serbia, University of Kragujevac
3
Institute for Animal Husbandry, Autoput 16, 11080, Belgrade-Zemun, Republic of Serbia
Corresponding author: genrad@agrif.bg.ac.rs
Original scientific paper
Abstract: In this study a relationship between the milk yield and following
reproduction traits: number of inseminations per conception [AIS], service period
[SP], age at calving [AC], calving interval [CI] and birth weight [BW] is studied.
The research involved dairy cows of European type of Black and White cattle
which are being intensively improved by the genes of Holstein – Friesian breed.
Mean values of general average for number of inseminations per conception,
service period, age at calving, calving interval and birth weight are: 2.00; 98.78;
1088.08; 376.70 and 36.40, respectively. In general, the estimated phenotypic
correlations (rp) between the milk yield over standard lactations and fertility traits
were positive and unfavorable, and ranged from 0.24 for the relationship between
milk yield and birth weight to 0.70 for relationship between milk yield and age at
calving.
Key words: level of milk yield, reproduction traits, phenotypic correlation, Black
and White cattle
Introduction
In intensive milk production over a longer period of years the problem
regarding the reproduction and diminished fertility in cows has been observed. For
a practical raising of cattle it is important to study the influence of the level of milk
yield on bovine fertility since between reproduction and lactation there is a close
physiological correlation. By studying the influence of milk yield on bovine
fertility different results have been obtained. The authors of a great number of
papers have reported that high-yielding dairy cows exhibit lower fertility and
greater health disorders in relation to low producing dairy cows (Rauw et al.,

488
R. Djedović et al.
1998; Pryce et al.,1999, 2004; Sandoe et al., 1999; Lucy, 2001; Rokstrom et
al.,2001; Windig et al., 2005; Tekerli and Koçak, 2009; Scholz et al., 2010; Sun et
al., 2010).
These problems are present especially on those farms that raise larger
populations of high-producing Holstein and Black and White dairy cows (Castillo-
Juarez et al., 2000; et al Lucy, 2001; Sørensen and Ruiz, 2007) who are also
expected, besides producing high milk yield in lactation, to produce a healthy and
vital calf once a year. If a disorder occurs in bovine reproductive cycle, besides its
direct consequences on realized milk production, the realization of normal herd
maintenance is also rendered impossible.
As a confirmation of a very complicated processes in bovine reproduction
serves a record which shows that only about 40-50% cows become pregnant at first
insemination (Royal et al,. 2000), thus the fertility in the populations of American
Holstein cows is decreasing at the rate of 0.5 -1.0% units annually (Bousquet et al.,
2004).
Taking into account that economic importance of reproductive traits of
dairy cattle is essential it is necessary to understand the factors which influence
bovine fertility, and they are all divided into genetic and paragenetic
(environmental factors). The success of improving fertility by genetic means is
limited by low heritability performance although it does not mean that genes do not
affect this trait but more that the value of additive genetic variation is small
(Djedović et al., 2002). The fertility in cows is considerably more influenced by
environmental factors because the majority of variations of reproductive
parameters is under their influence. When single factors are analyzed it is observed
that their influence on bovine fertility is of a different degree and also that a
complex influence is possible. By the entire understanding of more different effects
on reproductive traits in cows the fertility of dairy herds can be improved.
The research of some authors (Rokstrom et al,. 2001; Windig et al., 2006)
showed that there are unfavourable correlations between milk yield and fertility
traits. They have confirmed low to mean values of coefficients of phenotypic and
genetic correlations between milk yield and fertility traits (Pryce et al., 1999;
Oltenacu and Algers, 2005; Jagusiak, 2006).
Antagonistic relationship between these two groups of traits confirmed in
literature shows the necessity that the fertility traits should be included in overall
selection index.
The aim of this paper was to establish the effect of different levels of milk
yield on the most important reproduction traits as well as to give the grounds for
the estimation of genetic variability and relationship between studied traits
depending on the level of milk yield.

Material and methods
The effect of the level of milk ...
Data
The research on the effect of different level of milk yield as a paragenetic
factor on the reproduction traits was conducted on a farm of high-yielding Black
and White cattle. The farm is situated in the region characterized by intensive
vegetable production with capacity of about 500 milking animals. The cows belong
to European type of Black and White cattle whose genotype is being improved
intensively by introducing genes of Holstein-Friesian breed (HF). The study
included 730 standard lactations in a whole.
The criteria for involving the animals into trial were as follows:
- to be involved in milk production during the trial period,
- to have records on origin for three preceding generations,
- to have concluded the first, second and third lactation,
- to have lactations not shorter than 210 days.
For study purposes, that is, for establishing the relationship between the level
of milk yield and reproduction traits all the cows were assigned per lactations into
three groups with different share of Holstein genes. The groups were formed based
on the distribution of the frequency of genotypes regarded per lactations. All the
cows per lactation groups were classified into three classes depending on the level
of milk production (I, II and III). The milk yield classes were formed on the basis
of distribution of frequency of milk yield regarded per lactations (Table 1).
An average milk yield for the first (I) level was 5786, the second (II) 6976 and
the third (III) 8263 kg.
Table 1. Distribution of records per lactations, share of HF genes and milk yield levels
Share of HF genes, % n
Lactation I II III I II III
I 0-33 34-48 49-100 87 118 104
II 0-33 34-48 49-100 75 100 61
III 0-33 34-48 49-100 54 85 46
Level of milk yield, kg
Lactation I II III I II III
I 5500 91 141 77
II 6200 73 89 74
III 6700 56 72 57
I-III 6100 220 302 208
Model
Phenotypic variability and correlation of studied traits were established by
the Least Square Method (Harvey, 1990) that is, by the use of following fixed
model:
489

490
R. Djedović et al.
Yijkl = μ + Li +Nj +LNij +Gk +Sl + LSil + b1 (x1- 1) + b2 (x2- 2 ) + eijkl
where:
Yijklmo – is the manifestation of the trait of o- individual, i- lactation, j- milk yield
level , k- year of calving, l – calving season,
μ – general average of population for given trait,
Li – fixed effect of i - lactation (i= 1,..3),
Nj – fixed effect of j – milk yield level (j= 1,..3),
LNij – fixed effect of i – lactation and j - milk yield level (ij = 1,..9),
Gk- fixed effect of k- year at calving (k= 1,..5),
Sl – fixed effect of l – calving season (l= 1,..4),
LSil – fixed effect of interaction of i – lactation and fixed effect of l – calving
season (il=1...12),
b1 – linear regression effect of the share of Holstein-Friesian (HF) bovine genes,
b2 – partial regression effects of the share of Holstein-Friesian (HF) genes per milk
yield levels,
eijkl – random error with characteristics N (0, σ 2 ).
Results and Discussion
The mean values as well as the deviations and deviation errors from the
mean value for studied reproduction traits depending on the level (classes) of milk
yield are shown in Table 2.
Table 2. Mean values (LSM), mean deviations (q) and standard errors of mean deviations (S q)
for reproduction traits depending on milk yield level 1
Level of
Reproduction traits
milk yield n AIS SP AC CI BW
LSM
Se
730
2.00
0,07
98,78
2,71
1088,08
0,92
376,70
2,68
36,40
0,15
I Low
q
Sq
220
-0,32
0,08
-14,52
2,78
-1,56
0,94
-14,23
2,76
-0,28
0,15
II
Medium
q
Sq 302
-0,12
0,07
-7,39
2,54
0,27
0,86
-7,32
2,52
0,09
0,14
q
0,44 21,91 1,29 21,55 0,19
III High
208
Sq
0,08 2,79 0,95 2,77 0,15
Fexp 13,80 ** 36,90 ** 2,73 N.S . 36,18 ** 2,56 N.S .
1
LSM = Least squares means, SE = Standard error; AIS= number of inseminations per conception;
SP= service period (days); AC= age at calving (days); CI= calving interval (days); BW= birth weight
(kg)
NS – (P>0,05) * - (P

The effect of the level of milk ...
Mean value of the number of inseminations per conception for all animals
was 2.00±0.07. Deviations from the mean value ranged in the interval from –0.32
(I class) to 0.44 (III class). By the variance analysis a statistically highly significant
differences (P0,05) * - (P

492
R. Djedović et al.
three classes a highly significant correlation was confirmed. The correlation
coefficients were rp=0.38 ; rp=0.39 and rp=0.46 (I, II and III level).
In animals with low milk production (I level of milk yield) a poor, positive
and highly significant correlation between the milk yield in standard lactation and
calving interval was confirmed. On the contrary, both in cows with mean and high
milk yield (II and III class) a mean, positive and highly significant correlation was
confirmed. The phenotypic correlation coefficients were for the first class rp= 0.38 ;
second rp=0.41 and third rp= 0.48.
In relation to previously mentioned traits, phenotypic correlation between
the milk yield and the age at calving was moderately strong and positive whilst per
levels of milk yield it was rp= 0. 70 (I level); rp=0.69 (II level) and rp=0.48 (III
level).
For I and II level of milk yield a phenotypic correlation between the milk
yield and birth weight was mean, positive and highly significant. In the third level of
milk yield a weak but also positive and highly significant correlation was observed.
By increasing the milk yield a number of inseminations per successful
conception increased as well. Likewise the previous trait a symmetrical increase of
prolonged service period depending on the increase of milk yield was also
established. In the trait regarding the age of cow at calving a high variability was
not confirmed, contrary to previous traits. By increasing the milk yield a trend of
increasing the duration of calving interval was manifested. Deviations in birth
weight from the average value are rather low, so that the observed variations occurred
most probably under the effect of some factors not studied herein.
By analysing the established values of reproductive traits it is observed that
the effect of the level of milk yield on greater number of the same traits was
significant (P

The effect of the level of milk ...
Estimated phenotypic correlations are unfavourable in general since high-yielding
cows display weaker reproductive efficiency.
The greatest number of estimated values of phenotypic correlations
between the milk yield and reproductive traits published in literature are on the low
and mean level. Price et al. (1999) estimate that unfavourable correlation between
the milk yield and calving interval in the populations of dairy cattle was (0.28 ±
0.06). Hansen et al. (1983) estimated that correlation between the milk yield and
number of inseminations per conception was 0.37.
Number of service periods is the most important part of the calving interval,
because variation in gestation length, which represents the second part of the
calving interval, is very low (Jagusiak, 2006). Therefore, estimates of the
phenotypic correlations between calving interval and milk yield traits are usually
similar to those between service period and yield traits.
Thus, Veerkamp et al. (2001) report that estimated correlations between the
milk yield and reproductive traits were low and that the values for calving interval
were 0.19; for service period 0.20; and for days to first service 0.15.
Similarly to our study Marti and Funk (1994) studied the correlation between the
milk yield at different levels and reproduction traits and concluded that the highestproducing
cows had the longest service period.
Contrary to all previously cited authors Raheja et al. (1989) and Pantelić
et al. (2008) established the lowest coefficients of phenotypic correlations between
the milk yield and reproductive traits.
Conclusion
The unfavourable phenotypic correlations obtained between the milk yield
and fertility traits in this study as well as the results of cited authors show that the
choice of cows based only on the production of milk, over a longer time period,
may lead to diminishing of the bovine fertility. Therefore, when estimating the
bovine breeding value the values of phenotypic and genetic correlations between
the traits of milk yield and fertility should be included in an entire selection index.
Uticaj nivoa mlečnosti na osobine plodnosti krava crno bele
rase
R. Đedović , V. Bogdanović, G. Trifunović, M. Petrović, M. M. Petrović, D.
Stanojević
Rezime
U radu je ispitivan odnos između prinosa mleka i sledećih osobina plodnosti:
indeks osemenjavanja [IO], servis period [SP], uzrast pri teljenju [UT],
493

494
R. Djedović et al.
međutelidbeni interval [MI] i masa teladi pri rođenju [MTR]. Ispitivanja su se
odnosila na mlečne krave evropskog tipa crno-belih goveda koje se intenzivno
oplemenjuju genima holštajn-frizijske rase. Za analizu je korišćen Metod najmanjih
kvadrata – LSMLMW (HARVEY, 1990), fiksni model.
Srednje vrednosti opšteg proseka za indeks osemenjavanja, servis period, uzrast
pri teljenju, međutelidbeni interval i masu teladi pri rođenju su: 2,00; 98,78;
1088,08 ; 376,70 i 36,40, respektivno.
Procenjene fenotipske korelacije (rp) između prinosa mleka i osobina plodnosti
generalno su bile pozitivne i nepovoljne, i imale su vrednosti od 0,24 za odnos
između prinosa mleka i masu teladi pri rođenju do 0,70 za odnos između prinosa
mleka i uzrasta pri teljenju.
References
AMANI Z., GADER A., KHAIR M., LUTFI A., PETERS K. (2007): Milk yield
and reproductive performance of Friesian cows under Sudan tropical conditions.
Arch Tierz 50, 155-164.
ATIL H. (2000): Genetic relationship between days open and days dry with milk
yield in a herd of Holstein Friesian cattle. Arch Tierz 43, 583-590.
BOUSQUET D., BOUCHARD E., DUTREMBLAY D. (2004): Decreasing
fertility in dairy cows: Myth or reality? Proc. 23 rd World Buiatrics Congress,
Quebec, Canada, 6.
CASTILLO-JUAREZ H., OLTENACU P.A., BLAKE R.W.,
MCCULLOCH C.E., CIENFUEGOS-RIVAS E.G. (2000): Effect of herd
environment on the genetic and phenotypic relationships among milk yield,
conception rate, and somatic cell score in Holstein cattle. J Dairy Sci 83,
807–814.
DEMATAWEWA C.M., BERGER P.J. ( 1998): Genetic and phenotypic
parameters for 305-day yield, fertility, and survival in Holsteins. J Dairy Sci
81, 2700-2709.
DJEDOVIĆ R., LATINOVIĆ D., STOJIĆ P., BOGDANOVIĆ V.,
TRIFUNOVIĆ G. (2002): Heritability of milk traits in cattle depending on
the level of production. Biotechnology in animal husbandry 18, 17-22.
HANSEN L.B., FREEMAN A.E., BERGER P.J .(1983): Yield and fertility
relationships in dairy cattle. J Dairy Sci 66, 293-305.
HARVEY, WR (1990): Least-Squares and Maximum Likelihood Computer
Program.

The effect of the level of milk ...
JAGUSIAK W. (2006): Fertility measures in Polish Black-and-White cattle. 3.
Phenotypic and genetic correlations between fertility measures and milk production
traits. J Anim Feed Sci 15, 371–380.
LUCY M.C. (2001): Reproductive loss in high-producing dairy cattle:
Where will it end? J Dairy Sci 84, 1277-1293.
MARTI C.F,. FUNK D.A. (1994): Relationship between production and
days open at different levels of herd production. J Dairy Sci 77, 1682-1690.
M’HAMDI N., ALOULOU R., BRAR S. K., BOUALLEGUE M., M. BEN
HAMOUDA M. (2010): Phenotypic and genetic parameters of reproductive
traits in tunisian holstein cows. Biotechnology in Animal Husbandry 26 (5-
6), 297-307.
OJANGO J.M., POLLOTT G.E. (2001): Genetics of milk yield and fertility
traits in Holstein-Friesian cattle on large-scale Kenyan farms. J Anim Sci
79, 1742-1750.
OLTENACU P.A., ALGERS B. (2005): Selection for increased production
and the welfare of dairy cows. Ambio 34, 311-315.
PANTELIĆ V., PETROVIĆ M.M., ALEKSIĆ S., SRETENOVIĆ LJ., OSTOJIĆ-
ANDRIĆ, NOVAKOVIĆ Ž. (2008): Investigation of the genetic correlation
between milk and fertility traits of first calving cows of simmental breed.
Biotechnology in Animal Husbandry 24 (5-6), 1-8.
PRYCE J.E., NIELSEN B.L., VEERKAMP R.F., SIMM G. (1999): Genotype and
feeding system effects and interactions for health and fertility traits in dairy cattle.
Livest Prod Sci 57, 193–201.
PRYCE J.E., ROYAL M.D., GARNSWORTHY P.C., MAO I.L. (2004): Fertility
in the high-producing dairy cow. Livest Prod Sci 86, 125–135.
RAHEJA K., BURNSIDE E., SCHAEFFER L. (1989): Relationships Between
Fertility and Production in Holstein Dairy Cattle in Different Lactations. J Dairy
Sci 72, 2670-2678.
RAUW W.M., KANIS E., NOORDHUIZEN-STASSEN E.N., GROMMERS F.J.
(1998): Undesirable side effects of selection for high production efficiency in farm
animals. Livest Prod Sci 56,15–33.
ROXSTROM A., STRANDBERG E., BERGLUND B., EMANUELSON U.,
PHILIPSSON J. (2001): Genetic and environmental correlations among female
fertility traits and milk production in different parities of Swedish red and white
dairy cattle. Acta Agric Scand 51,7–14.
ROYAL M.D., DARWASH A.O., FLINT A.P.F., WEBB R., WOOLIAMS
J.A., LAMMING G.E. (2000): Declining fertility in dairy cattle: changes in
traditional and endocrine parameters of fertility. Anim Sci 70, 487-501.
495

496
R. Djedović et al.
SANDOE P.B., NIELSEN L., CHRISTENSEN L.G., SORENSEN P.
(1999): Staying good while playing God-The ethics of breeding farm
animals. Anim Welf 8,313–328.
SCHOLZ H., BEYER B., ANACKER G., WÄHNER M. (2010):
Relationship between milk yield, fertility and amount of disease of Holstein
Friesian dairy cows [in German]. Arch Tierz 53, 283-292.
SØRENSEN A.C., LAWLOR T., RUIZ F. (2007): A survey on fertility in
the Holstein populations of the world. Proc. Int Conf on Fertility in dairy
cows, Liverpool Hope University, UK (EAAP Satellite Meeting), 17.
STEVENSON J.S. (1999): Can you have good reproduction and high milk
yield? Hoard's Dairyman 144–536.
SUN C., MADSEN P., LUND M.S., ZHANG Y., NIELSEN U.S., SU G.
(2010): Improvement in genetic evaluation of female fertility in dairy cattle
using multiple-trait models including milk production traits. J Anim Sci 88,
871–878.
TEKERLI M., KOCAK S. (2009): Relationships between production and
fertility traits in first lactation and life time performances of Holstein cows
under subtropical condition. Arch Tierz 52, 364-370.
VACEK M., STADNIK L., ŠTIPKOVA M. (2007): Relationships between
the incidence of health disorders and the reproduction traits of Holstein
cows in the Czech Republic. Czech J Anim Sci 52, 227–235.
VEERKAMP R.F., KOENEN E.P.C., DE JONG G. (2001): Correlations
among body condition score, yield, and fertility in first-parity cows
estimated by random regression models. J Dairy Sci 84, 2327-2335.
WINDIG J.J., CALUS M.P.L., VEERKAMP R.F. (2005): Influence of Herd
Environment on Health and Fertility and Their Relationship with Milk
Production. J Dairy Sci 88, 335–347.
WINDIG J.J., CALUS M.P.L., BEERDA B., VEERKAMP R.F. (2006):
Genetic Correlations Between Milk Production and Health and Fertility
Depending on Herd Environment. J Dairy Sci 89, 1765-1775.
Received 11 July 2012; accepted for publication 20 August 2012

Biotechnology in Animal Husbandry 28 (3), p 497-507 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.237
DOI: 10.2298/BAH1203497N
RESULTS OF THE BIOLOGICAL TEST OF
SIMMENTAL BULLS IN CENTRAL SERBIA
D. Nikšić 1 , V. Pantelić 1 , D. Ostojić-Andrić 1 , P. Perišić 2 , M. Petričević 1 , R.
Đedović 2 , M. Lazarević 1
1 Institute for Animal Husbandry, Autoput 16, P. Box 23, 11080, Belgrade-Zemun, Republic of Serbia
2 Faculty of Agriculture University of Belgrade, Nemanjina 6, 11080 Belgrade, Republic of Serbia
Corresponding author: draganniksic84@gmail.com
Original scientific paper
Abstract: Exceptionally important aspect in cattle production, from
the aspect of production and economy, is ensuring normal and regular
fertility. Every cattle breeder wants to have high-yielding animals which at the
same time have good fertility. In proper cattle breeding this means that from each
cow during single year one healthy calf is obtained. Use of artificial insemination
has enabled that one breeding male is used as sire for several tens of thousands of
progeny, however, there is always the risk that sires could be carriers of lethal and
semi-lethal genes, which can cause huge losses of calves. In order to bring these
undesirable occurences to a minimum, so called bilogical test is included in the
breeding and selection activities, i.e. bulls are tested through their progeny for
presence of difficult calving (dystocia) abd calf losses caused by genetic anomalies.
In Republic of Serbia, this test is carried out on ehtire Simmental population in an
exchange of data produced by primary breeding organizations. Per each tested bull,
it is necessary to have data on minimum 50 randomly selected calves. Calves are
examined visually 65 days after the birth, at the latest. In the present study, the
results of the biological testing of 35 Simmental bulls, sires of total 3572 calves on
the territory of Central Serbia, in the period 2008-2009, are used. The effect of
sires on parameters of biological test were studied: body mass of calves at birth,
calf score and calving score. Also, bulls were ranked based on said parameters,
male/female calves ratio and percentage of twins and still-born calves for each bull
individually were determined. After the rank of bulls was established, the
correlation between the rank and studied fertility traits was determined.
Correlations were expressed using the Spearman coefficient. Research results show
that the effect of bulls was very high p

498
Introduction
D. Nikšić et al.
For present trends in modern production of milk and meat it is absolutely
unacceptable to have one-sided selection for productive traits only, because it can
greatly contribute to numerous metabolic disorders and shortening of productive
life. Modern/contemporary breeding goals and programs are focused primarily on
reaching and maintaining of the optimal balance between productive and functional
traits without any health risk and compromising of animals’ ethological
characteristics (Ostojić-Andrić et al., 2011). Regardless of the breeding goal and
defined program, fertility and fitness traits must be iontegrated, i.e. traits of
resistance to diseases (Trivunović et al., 2011).
Biological test provides the right information on breeding value of bulls in
the progeny testing. It includes study of bulls in terms of inheritance of
degenerative properties, potential incidence of calves with congenital deficiencies
and defects, lethal and semi-lethal factors which ensures acurate insight of the
quality of calves obtained from individual bull sire. With this test, practically, it is
established if breeding male is carrier of lethal, semi-lethal or harmful genes,
where:
• Lethal genes cause death of the progeny during
gestation/gravidity, during or immediately post- parturition;
• Semi-lethal genes cause birth of avital progeny which dies shortly
after birth, or some time subsequent to parturition;
• Harmful genes which do not cause directly the death of calves, but
induce diminishing of the vitality and resistance of progeny of harmful genes.
Biological test starts at the moment when young bulls collect the first 750
doses of semen. In that period, according to pre-defined schedule, semen is
distributed to farms and cows and heifers are inseminated randomly Đedović
(2004).
Biological test of young bulls is carried out in production conditions of the
population. In order to establish if potential breeding male is carrier of undesirable
genes (lethal, semi-lethal or harmful genes), it is necessary to study at least 50
calves. Calves are examined in the first month subsequent to parturition (at latest
65 days post partum), and focus is usually on 10 defects, and which of them will be
included in the test depends on the specific population.
Fertility is almost regularly the first limiting factor in milk production. One
of the most important reproductive traits is body mass of calves at birth. It is
considered as one of the major traits which influence the calving progress, when
there is objective risk of incidence of difficult calving (dystocia) and calf mortality
Johanson and Berger, (2003); Linden et al., (2009). In case of difficult calving
(dystocia), the main issue is disproportion of the fetus size and birth canals of the
dam, which is most often case with first calving females, as stated by Martinez et

Results of the biological...
al.,(1983) and Lombard et al. (2007). The expression of this trait is also influenced
by hereditary and environment factors, the most important are the following: breed,
calf sex/gender, calving order, duration of gravidity, dam’s age, duration of dry
period, dam’s body mass, breeding/rearing and nutrition as stated by Latinović et
al.,(1983); Skalicki et al., (1991); Trifunović et al. (1998).
In the present study production results of Simmental bulls are analyzed and
cow fertility results in period 2008 – 2009 presented, in order to present to
scientific community the genetic potential of Serbia's cattle population in terms of
productivity of bulls and fertility of cows of Simmental breed and also to indicate
the frequency of incidence of still born calves and calves with anomalies.
Materials and Methods
The data recorded by the Main Breeding Organization – Institute for
Animal husbandry, Belgrade-Zemun, was used in the present study. Biological test
was done on 35 bulls of Simmental breed with data on 3.572 calves, born in the
period from 2008 to 2009 on the territory of Central Serbia. Analysis included
following traits: body mass at birth (BMB), calf score at birth (CSB), calving score
(CS), type of birth (TB), number of still born calves (SBC) and sex ratio.
Frequencey, i.e. percentage values were determined for calf score at birth,
calving score, type of birth and number of still born calves. The effect of sires on
variability of traits (body mass of calves, calf score at birth, calving score)
observed in the biological test was determined using single factor variance analysis
(p

500
D. Nikšić et al.
Novaković et al. (2012) in the study of the effect of numerous systematic factors on
fertility parameters of the biological test. In the same study, the presence of
significant variability in regard to body mass at birth between sires was established.
Table 1. Mean values and variability of traits in the biological test
Traits x min max SD CV Se
Body mass at birth 44,54 25 73 5,27 11,03 0,09
Calf score at birth 4,53 2 5 0,59 13,02 0,01
Calving score 4,66 2 5 0,55 11,8 0,01
In the analysis of variance it was established that all three traits in the
biological test vary significantly (p

Results of the biological...
Table 3. Rank of bulls for studied fertility traits based on average values
Bull
name
Bull
HB
Body mass
at birth
Rank
Calf
score at
birth
Rank
Calving
score
DIETER 1427 44,74 14 4,68 18 4,77 20
STIER 1492 43,99 19 4,28 24 4,78 19
DONER B158 42,33 30 4,25 25 4,62 26
HUMSTREIF B206 43,02 24 3,95 32 5 1
WEHOR B213 42,93 26 3,91 33 5 2
ERFAS 1352 54,2 1 5 1 5 3
FAO B19 42,99 25 4,13 27 4,09 32
FEST B145 43,8 21 4,14 26 4,05 33
HOFAL 1500 43,28 23 4,12 30 4,03 34
ZASTRADA 1453 49,25 4 4,8 16 4,75 22
WEIPORT V026 50,79 2 4,93 10 4,55 28
HORMAZ 1488 48,36 7 4,56 21 4,79 17
HORMIT 1470 49,6 3 4,52 22 4,57 27
VALBO B279 44,38 15 4,64 19 4,86 12
ZAHLO 1497 45,54 10 4,63 20 4,66 25
RAUBECK 1507 44,2 18 3,81 34 4,99 4
WOZ 1433 42,09 31 3,6 35 4,87 10
HODAR B68 43,35 22 4,07 31 3,99 35
RADEX B51 43,97 20 4,13 28 4,12 30
RAŠA V626 48,95 5 4,76 17 4,87 11
VALIAN 1501 48,89 6 4,97 4 4,97 5
PORTO B118 42,51 28 4,87 13 4,92 7
FEBRO B50 46,17 9 4,83 15 4,77 21
HODBAL B275 44,96 12 4,94 9 4,79 18
DION B244 44,32 17 4,96 6 4,81 15
MOOR 1504 44,83 13 4,95 7 4,84 13
MARNOLD 1508 44,35 16 4,95 8 4,84 14
HORAU 1411 40,43 33 5 2 4,67 24
MOBI B112 40,35 34 4,9 12 4,74 23
LEO B164 39,61 35 4,34 23 4,27 29
REPRON 1479 45,28 11 4,92 11 4,92 8
MARMAL B333 42,68 27 4,98 3 4,9 9
MALINT B232 47,78 8 4,13 29 4,12 31
MILAN V1023 41,21 32 4,84 14 4,8 16
VEBER B307 42,47 29 4,97 5 4,97 6
501
Rank

502
D. Nikšić et al.
values of body mass of calves, calf scores and calving scores. The best
ranked bulls are Erfas HB 1352 for body mass at birth and calf score at birth, and
for calving score the best ranked bull is Humstreif HB B206.
Subsequent to ranking of bulls, the level of concurrence between obtained
ranks was tested. Table 4 presents correlation coefficients expressed by Spearman
coefficient. Between bull ranks weak and positive correlation was determined in
the range from 0,042 to 0,285, which enables carrying out of direct selection. It is
particularly important that the correlation is positive, it means that highly ranked
bull in regard to calving score can produce calves with acceptable body mass at
birth.
Table 4. Correlations of bull ranks
Rank Rank correlation
BMB CSB 0,202**
BMB CS 0,042**
CSB CS 0,285**
In this study, many parameters of the biological test were studied for each
bull individually (Table 5). By comparing these results it can be established that the
total percentage of still born calves was significantly lower (1,23%) compared to
results obtained by Gottschalk et al. (1983). Considering that bulls Zahlo HB 1497
and Woz HB 1433 had more than 5 % of still born calves they were negative in the
biological test, and as such excluded from breeding.
One of the most important factors influencing the change in the gene
frequency of population is the bull semen used in artificial insemination. Therefore
it is very important to establish for each bull the presence of genes which influence
the equal sex ratio of the progeny. In conditions of artificial insemination, bulls
which produce approximately 50% of male and 50% of female offspring are
prefered. According to data used in the present study (Table 5), it was calculated
that in average more female calves (54,16%) are born than males (45,84%).
In the present study also the type of birth was determined (Table 5). Type
of birth means number of calves born per one parturition, i.e. if the result of calving
was single calf, twins or triplets. Of total number of calves 3752, there were no
triplets, and number of twins was 170 which is approximately 4,76%,
Also, the percentage values i.e. presence of calving scores for each bull
individually were calculated. Number of calves born by Cesarean cut was
exceptionally low – 5. Dams which underwent the Cesarean cut were inseminated
with semen of bulls Mobi HB B112 (2), Febro B50, Marmal B333 and Milan
V1023 (Table 5).
Since it is known that difficult calving (dystocia) has equaly adverse effect
on production of milk and meat, and knowing that body mass of calves at birth is

Results of the biological...
highly hereditary, bull sires with genes for high body mass of calves should not be
used for insemination of heifers and cows of easier type.
Table 5. Fertility parameters for each individual bull
Bull name
Bull
HB
Calving score Comments
Number
of
calves 5 4 3 2 MR BL
Sex ratio
(m-f)
1 DIETER 1427 101 87 5 9 - 4 10 39-62
2 STIER 1492 96 80 11 5 - 2 4 32-64
3 DONER B158 97 74 9 14 - 4 12 50-47
4 HUMSTREIF 206B 116 116 - - - - - 47-69
5 WEHOR 213B 109 108 - - - - - 57-52
6 ERFAS 1352 95 95 - - - - 10 49-46
7 FAO B19 92 8 84 - - - 6 39-53
8 FEST B145 85 4 81 - - - 3 34-51
9 HOFAL 1500 249 10 236 3 - 1 7 115-134
10 ZASTRADA 1453 81 61 20 - - - 6 43-38
11 WEIPORT V026 94 52 42 - - - 10 25-69
12 HORMAZ 1488 81 69 7 5 - - - 34-47
13 HORMIT 1470 92 65 14 13 - - - 48-44
14 VALBO B 279 100 86 14 - - 2 10 46-54
15 ZAHLO 1497 95 75 8 12 - 9 8 47-48
16 RAUBECK 1507 85 84 1 - - - 2 43-42
17 WOZ 1433 92 85 2 5 - 5 - 45-47
18 HODAR B68 89 1 86 2 - 1 6 42-47
19 RADEX B51 89 11 78 - - - 8 48-41
20 RAŠA V626 87 76 11 - - - - 34-53
21 VALIAN 1501 91 89 1 1 - 2 12 31-60
22 PORTO B118 111 103 7 1 - 5 14 34-77
23 FEBRO 50 81 65 14 1 1 1 - 46-35
24 HODBAL B275 89 70 19 - - 1 4 49-40
25 DION B244 100 81 19 - - - 4 53-47
26 MOOR 1504 81 68 13 - - - 2 40-41
27 MARNOLD 1508 80 67 13 - - - 4 44-36
28 HORAU 1411 96 72 16 8 - - 2 39-57
29 MOBI B112 117 95 15 5 2 2 2 60-57
30 LEO B164 128 53 57 18 - - 2 69-59
31 REPRON 1479 172 159 13 - - 2 - 71-101
32 MARMAL B333 126 116 9 - 1 2 2 57-69
33 MALINT B232 93 26 52 15 - - 2 45-48
34 MILAN V1023 92 76 15 - 1 1 14 43-49
35 VEBER B307 91 88 3 - - - 4 40-51
In Table 6, the number and percentage of calves for each individual trait
are presented. By observing the results related to calf score and low incidence of
503

504
D. Nikšić et al.
anomalies, most of calves were scored 5 and 4, or around 96%. Obtained results
are consistent witjh results obtained by Đedović 2004., whereas results related to
calving score were consistent with results obtained by H. Karb et al. (1988).
Table 6. Number and percentage of calves according to individual scores
Calf
scores
Number
of calves
Percentage
of calves
Calving score
Number
of calves
Percentage
of calves
5 2069 57,93 5 2475 69,29
4 1364 38,18 4 975 27
3 121 3,39 3 117 3,28
2 18 0,5 2 5 0,14
Conclusion
In this study, the effect of bull sires on certain fertility parameters in
biological test was determined. Also, results for each bull individually were
determined.
Based on conducted research and obtained results it can be concluded that
the number of bulls who gave still born or calves with anomalies was very low,
whereas the number of calves receiving the highest scores was exceptionally high.
Sex ratio was close to the ideal 50%:50%, and percentage of twin births was close
to 5.
The right choice of bulls as future breeding animals can be way to improve
fertility traits and calving type. The use of tested bulls which are simultaneously
tested for milk traits, milking traits and body development traits as well as presence
of congenital anomalies and defects will influence the reduction of frequency of
undesirable genes in populations of Simmental cattle.
In this study, 35 bulls were ranked based on the average values of traits in
the biological test.
Bull ranking is important for selection experts and breeders because the
right selection of bull enables faster and more efficient realization of the breeding
goal. The choice depends on the primary operation of the breeder, who by viewing
the ranking of bulls can easily select adequate bull. The right choice of bull is also
imposed by the fact that in conditions of artificial insemination bull have
considerably higher intensity of fertility than cows.
Anaysis of obtained results and adequate distribution of semen of tested
and positvely scored and evaluated bulls in the biological test can contribute to
expression of positive results in improvement of fertility traits. Considering that
bulls have statistically influenced traits observed in the biological test, with rught
selection of future breeding sires it is possible to improve fertility traits and
suppress undesirable genes.

Results of the biological...
In view of all stated in the present study, the research of the evaluation of
the breeding value of cattle should be continued and also all known data associated
with the inherent variability of fertilit traits should be included, since in this way
the long term increase of the effects of applied selection in studied cattle selection
will be realized.
Acknowledgment
Research was financed by the Ministry of Education, Science and
Technological Development of the Republic of Serbia, project TR31053.
Rezultati biološkog testa bikova simentalske rase u
Centralnoj Srbiji
D. Nikšić, V. Pantelić, D. Ostojić-Andrić, P. Perišić, M. Petričević, R. Đedović, M.
Lazarević
Rezime
Izuzetno važan aspekt u govedarskoj proizvodnji, gledano sa proizvodnog i
ekonomskog stanovišta je obezbeđivanje normalne i redovne plodnosti. Svaki
odgajivač goveda želi da ima grla koja su visoko proizvodna i istovremeno
poseduju dobru plodnost. Za pravilno odgajivanje goveda to znači da se od svake
krave u toku godine dobije po jedno zdravo tele. Upotreba veštačkog
osemenjavanja omogućila je da jedan priplodnjak bude otac nekoliko desetina
hiljada potomaka. Međutim, uvek postoji rizik da su očevi nosioci letalnih i
semiletalnih gena, što može prouzrokovati velike gubitke teladi. Da bi se ove
nepoželjne pojave svele na najmanju moguću meru u odgajivački i selekcijski rad
uključeno je ispitivanje bikova po potomstvu na teška teljenja i gubitke teladi
izazvane genetskim anomalijama, tzv. biološki test. Ovaj test se u Republici Srbiji
vrši jedinstveno za čitavu populaciju simentalske rase, razmenom podataka između
odgajivačkih organizacija. Po svakom biku koji se testira, neophodno je imati
podatke za najmanje 50 slučajno odabrane teladi. Vizualni pregled teladi se obavlja
najkasnije do 65 dana, od datuma rođenja teleta. U ovom istraživanju korišćeni su
rezultati biološkog testa 35 bikova simentalske rase koji su bili očevi ukupno 3572
teladi na teritoriji centralne Srbije u periodu 2008. i 2009. godine. U radu je
ispitivan uticaj očeva na parametre biološkog testa: telesnu masu teladi po rođenju,
ocenu teleta i ocenu toka teljenja. Takođe je izvršeno rangiranje bikova na ove
parametre, kao i utvrđivanje odnosa muške i ženske teladi, procenat bližnjenja kao
505

506
D. Nikšić et al.
i procenat mrtvorođene teladi za svakog bika posebno. Nakon utvrđenog ranga
bikova izvršena je korelacija ranga ispitivanih osobina plodnosti. Korelacije su
iskazane Spirmanovim koeficijentom. Rezultati istraživanja pokazali su da je uticaj
bikova izuzetno visok p

Results of the biological...
perspectives and Challenges of Sustainable Livestock production”Belgrade,
Republic of Serbia, 5-7 th October 2011. Biotechnology in Animal Husbandry vol
27, 3, Book 2, p.975-984
SKALICKI Z., LATINOVIĆ D., LAZAREVIĆ LJ., STOJIĆ P. (1991): Fenotipske
karakteristike reproduktivnih osobina crno-belih goveda sa različitom proporcijom
gena holštajn-frizijske rase. Zbornik radova poljoprivrednog fakulteta, radovi sa
VII naučnog skupa zootehničara Jugoslavije, 2, 33, Beograd.
STATISTICA FOR WINDOWS VERSION 7. StatSoft.Inc (2004), Oxford, UK
TRIFUNOVIĆ G., LAZAREVIĆ LJ., LATINOVIĆ D., STOJIĆ P.,
STEVANOVIĆ LJ. (1998): Analiza mlečnosti i plodnosti krava tokom prve tri
laktacije gajenih slobodnim sistemom držanja. Savremena poljoprivreda br. 1-2,
str.49-54. Novi Sad.
TRIVUNOVIĆ S., IVANOVIĆ D., KUČEVIĆ D., PANTELIĆ V., KORORA J.,
RADINOVIĆ M. (2011): Genetski parametri za pojavu teških teljenja i broj
mrtvorođene teladi u populaciji krava holštajn frizijske rase vojvodine,
Biotechnology in Animal Husbandry, Vol 27 , Beograd
Received 11 May 2012; accepted for publication 10 August 2012
507

Biotechnology in Animal Husbandry 28 (3), p 509-516 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.2
DOI: 10.2298/BAH1203509G
STUDY OF ANIMAL WELFARE STATUS IN DAIRY
COW HERDS IN HUNGARY
R.T. Gudaj 1 , E. Brydl 2 , J. Lehoczky 2 , I. Komlósi 1
1Centre for Agricultural and Applied Economics Science; Faculty of Agricultural and Food Sciences
and Environmental Management; University of Debrecen, 138 Böszörményi street; 4032 Debrecen;
Hungary
2 Department of Animal Hygiene, Herd Health and Veterinary Ethology, Szent István University
Faculty of Veterinary Science, H-1078 Budapest; István u. 2; Hungary
*Corresponding author: rgudaj@agr.unideb.hu
Original scientific paper
Abstract:Animal welfare is a hot topic among consumers, producers and
researches nowadays. The major welfare problems of dairy cows are mastitis,
lameness, and any conditions which lead to impaired reproduction, inability to
express normal behaviour, emergency physiological responses or injury. This paper
summarizes preliminary results of project taken in 27 Hungarian dairy farms
evaluating general animal welfare. The most important areas for improving
animals’ wellbeing are related to facilities and comfort of resting. Findings include
slippery floors, cows struggling laying and standing in cubicles. Other measures
include hair loss, hocks, neck rail injuries and number of thin cows (Body
Condition Score 1 and 2). Mouldy silage and low quality of other feedstuff was
also found. In conclusion, preliminary results confirm strong demand for
monitoring farms and discussions with managers and farmers about welfare
measures needed to be taken on farms immediately.
Key words: dairy welfare, dairy welfare assessment protocol, welfare
standard, multivariable modelling, herd health
Introduction
According to von Keyserlingk et al. (2009), there are 3 major concerns related to
animal welfare: if the animal is functioning well, feeling well, and if the animal is
able to live according to its nature. These arguments are not new, because
producers have always been (at least they should be) concerned about the
conditions of animals in their care. So far, animal welfare was understood as a lack
of hunger, thirst, injuries and illness. Recently, welfare aspects are also focusing on
discomfort, distress, fear, pain and normal behaviour, as a result of keeping animals
in usually non animal-friendly environments (Stull, et al., 2005).

510
R.T. Gudaj et al.
Companies dealing with dairy products are interested in welfare of animals to give
consumers demanded clear and trustworthy information. The attempt was already
taken for establishing an European standard for welfare assessment systems in
order to facilitate intra-European trade and marketing created by Welfare Quality
Project (Veissier and Evans, 2010). There are claims that welfare of farm animals
has a huge impact on market demands and there is still need for developing
science-based on-farm systems to assess the animal welfare status (Matthews and
Wassmann, 2003). Over the last three decades the number of publications about
animal welfare and animal wellbeing has increased significantly. Such an output
reflects the economic demand for that information globally.
There are many aspects which lead the science and the experience to the
conclusions that welfare of dairy cows might be compromised (Ostojić-Andrić et
al., 2011; Hristov et al., 2011a; Hristov et al., 2011b). The fact that cows are
housed is already limiting factor for their natural needs which they can express on
the pasture. Facilities created by humans are not always suitable for cows. Cows
are given sometimes not enough resources they demand to produce as much milk
as their potential allows. High standards of animal welfare have been and will
continue to be important to the dairy industry. It is already proven that for lactating
cows the reduction in milk yield is associated with various diseases (EFSA, 2009;
Kossaibat and Esslemont, 1997; Trevisi et al., 2006). Immune system activation
requires metabolic energy when cow is ill. That state results in reduced feed intake.
Resources may be limited and may be diverted into immune function rather than
milk production, growth, or reproduction.
Cattle are exposed to dirt, because of mud and faeces in the environment they are
kept. Phillips and Morris (2002) reported that animals, particularly when
introduced, are avoiding those areas if possible. Those conditions could be
considered as compromising animal welfare. Schukken et al. (1990) concluded that
hygiene of husbandry system is related to mastitis and digital dermatitis (Hristov et
al., 2011c). There is also relation between cleanliness and gastrointestinal problems
(Rodriguez-Lainz et al., 1996). The presence of mud and dirt my be irritant and
causing discomfort. The cleanliness of animals demonstrates quality of
environment and can be useful indicator of farm animals’ welfare.
Materials and methods
The study was performed on 27 Holstein-Friesian dairy farms in Hungary. The
selection was firstly created on a principle of searching for as different farms as
possible. Chosen farms were farms different in: ownership (private and

Study of animal welfare status ...
cooperative), size (from 56 to 850 dairy cows), husbandry systems (free stall, straw
yard), access to the pasture (yes, no), scraping system (automatic, tractor), age of
buildings (modern, old ones), number of animals per water troughs (12-120),
surface quality (1 – relatively dry, no holes and not slippery; 2 – wet or some holes
or slippery; 3 – wet, some holes and slippery). Data collection was created
according to already existed assessments (Whay, et al., 2003). The project,
however, needed the protocol to be established again, as in Hungarian conditions
some of the points of the protocol would simply not work (for example – access to
the pasture – which is not so popular). Protocol covered animal health, behaviour,
hygiene and feedstuff condition of all animals on the farm. Additionally, general
production data, facilities, lameness treatment and stockman attitude is measured
for dairy cows. Finally, the following characteristics were chosen: dirty flanks,
udder, hind limbs with size of dirt splash or plaque more than the size of the palm
of a hand. An average flight distance was measured by approaching the animal at a
speed of one step per second and a step length of approximately 60 cm with the
arm held overhand at an angle of approximately 45º from the body. Dull and
obviously sick cows, hollow rumen, bloated rumen, fat animals, hair loss, non-hock
injuries, hocks injuries, thin animals, neck rail injuries were noted if any of signs
were easily visible. Rising was characterised as either unrestricted (1), mildly
restricted (2) when they modified their rising behaviour to stand up comfortably,
seriously restricted (3) when they took time to stand up and often hit fittings when
rising and when they rose on to their forelimbs before their hind limbs (dog
sitting). During the visits cows were assessed regarding their locomotion score and
body condition score. Five-point scale locomotion score of dairy cattle was used.
The system developed by Sprecher et al. (1997) has understandable objective
descriptions of posture and gait for scoring. This also includes subdivisions
between sound and clinically lame cows. Cows were provided relatively dry, free
of obstacles, concrete surface. Cows which were found in the cubicles were given
few minutes to recover after standing up, so impact of muscle crump would not
affect cows’ locomotion. For evaluating body condition score 5-point scale
condition score of dairy cattle published in (Rodenburg, 2000) was used. For
measuring intra-observer variation notes were made at the beginning of the
observation. Cows walking were assessed and results were recorded. Half of the
cows were observed for the second time at the end of each visit and results were
compared with the first observation. Number of cows observed twice ranged from
5 (during the first visit) up to 60 (during the last visit, if no cow left the farm). In
average 83% repeatability of locomotion scores and 91% reputability in body
condition scores were estimated. Full animal welfare assessment with checking
every dairy cows’ locomotion took around 6-7 hours for 500-600 heads herd.
511

512
Results and Discussion
R.T. Gudaj et al.
Summary preliminary results of the general animal welfare measures taken are
shown in Table 1. These findings are similar to those of Grandin (2010). On the
other hand, this research will be significantly different to others. This research
showed that in older animals more welfare problems occur. In the study 27,6% of
older calves were found with dirty flanks, what emphasises the lack of proper
bedding. Dull and obviously sick and hollow rumen of dry cows (3.9% and 9.7%,
respectively) is just slightly different from those of milking cows. That means
milking cows are in such a poor condition or dry cows are in such a good state.
Heifers, what is natural, are more active and less trustful to humans what explains
why flight zone of these animals is the biggest. On the other hand there is a doubt if
heifers are in a good state before calving, because 8.7% of them had bloated
rumen, 22.5% of them were with Body Condition Score 4 and 5 and 37.8% had
dirty udders. Milking cows were found with most of the measures compromising
animal welfare. Rising, both in straw yards and in free stalls gave an average of 1.8
(where 1- unrestricted; 2- mildly restricted; 3- seriously restricted). This illustrates
that cubicles were not suitable for cows and straw yards which were poorly bedded.
The next three conditions – hair loss, non-hock injuries and hocks lesions (16.7%,
20.9% and 22.3%, respectively) were also found only in milking cows and reflect
poor cubicles’ design. Similar results were found by Potterton et al. (2011),
however, in that study moderate (25.6%) and severe (14.5%) hair loss was
measured. The same authors found only 6.7% cows with moderate hocks and only
2.5% with severe hocks lesions. Those results correspond with Lombard et al.
(2010) who estimated only 4.1% of cows in average with hocks lesions. However,
study of Rutherford et al. (2008) highlighted 49.1% of hock lesions. In the current
study 25.7% of cows were found with Body Condition Score 1+2 might manifest
some energy imbalances. 41.8% of cows with dirty hind limbs are as an effect of
lying in dirty areas which in fact were probably the only locations in a straw yard
the cows were able to take place. In free stall barns that high percentage of dirty
cows might be result of finding muck and manure in the alleys more comfortable
than boxes. Unlikely to this study, Lombard et al. (2010) evaluated only 8.8% of
cows being highly soiled. Lameness is linked to multivariable conditions ranging
from 5.7% to 50.9% with an average of 27.9%. Occurrence of lameness was found
to be similar to findings of Haskell et al. (2006), Huxley et al. (2004), and
Rutherford et al. (2009) with 19.3%, 24%, and 39% of cows found clinically lame
respectively. Neck rail injuries found in 61.7% of milking cows are related to
wrong positioned feed rail and/or feeding trough.

Study of animal welfare status ...
Table 1. Summary of the general animal welfare measures taken
Welfare measure
Younger
calves (0-70
days)
Older calves
(70+ days)
Dry cows Heifers
513
Milking
cows
Dirty flanks (%) 11.6 27.6 18.3 22.6 26.6
Dull/obviously sick (%) 3.2 2.7 3.9 3.0 3.7
Hollow rumen (%) 5.6 5.4 9.7 6.2 9.2
Average flight distance (m) 0.7 0.8 0.9 1.0 0.8
Bloated rumen (%) 2.0 4.7 6.5 8.7 3.7
Fat animals (BCS 4+5) (%) 4.9 4.8 16.0 22.5 16.1
Dirty Udder (Underneath) (%) 9.0 30.5 28.8 37.8 25.5
Rising (1/2/3) 1.1 1.4 1.6 1.7 1.8
Hair loss (%) 1.3 1.8 9.5 5.4 16.7
Nonhock injuries (%) 0.9 1.1 8.6 4.7 20.9
Hocks lesions (%) 0.9 5.3 17.0 7.8 22.3
Thin animals (BCS 1+2) (%) 12.0 5.6 15.1 6.6 25.7
Lameness (%) 0.4 2.5 21.6 6.7 27.9
Dirty hindlimbs (%) 8.8 34.8 28.5 39.7 41.8
Neck rail injuries (%) 1.1 43.9 61.1 39.6 61.7
Conclusion
There is an immediate need to point out farmers the most important welfare
problems on the farms. The points highlighted in the preliminary results confirm
laying and resting conditions are compromised. Comfort is very important in cows’
daily routine, because the animals need a time for relaxing, but that is highly
limited. Hungarian dairy enterprises (particularly those in project) lack in good
quality, modern barns. Most of them are refurbished old buildings which, in many
cases, even some changes were applied, still cannot meet cows’ needs. There is an
increasing interest of farm managers and farmers about dairy welfare. During visits
on farms, when each protocol was in progress or was finished, there has been a
short time for discussion with famers. Cows’ and farms’ conditions become easier
to imagine and understand when they are compared with other farms by showing
pictures and exact numbers (Grandin, 2010). This method of comparing welfare on
farms is useful in explaining, for example, neck rail injuries. Although, most of the
managers see the problem and they find wrong feeder or rail height, most of them

514
R.T. Gudaj et al.
blame cows that they are pushing the bar too hard to reach the food. As a counterargument
the protocol confirms that positions of the bar and the feeder where only
3-9% of cows are found as a cause of injuries on necks. The same positive
perception is achieved when pictures or examples of compromised welfare are
shown directly to the person. Welfare was checked before suggestions were given
and other independent improvements were made. Next step will be to check the
cows after a period of time, if changes helped to increase welfare standards or not.
Studija o statusu dobrobiti životinja u zapatima mlečnih
krava u Mađarskoj
R.T. Gudaj, E. Brydl, J. Lehoczky, I. Komlósi
Rezime
Dobrobit životinja je aktielna tema među potrošačima, proizvođačima i
istraživačima. Glavni problemi i pitanja u okviru dobrobiti životinja su mastitis,
hromost, kao i svi drugi uslovi koji dovode do poremećaja u reprodukciji,
nemogućnosti da životinje izraze svoje normalno ponašanje, i vanredne fiziološke
reakcije ili povrede. U ovom radu se daje prikaz preliminarnih rezultata projekta sa
27 farmi mlečnih goveda u Mađarskoj u oceni opšte dobrobiti životinja.
Najznačajniije oblasti za poboljšanje dobrobiti životinja se odnose na objekte i
udobnost životinja, komfor i mogućnost odmora. Nalazi istraživanja ukazuju na
klizave podove, krave koje sa poteškoćama leže odnosno stoje u boksovima,
gubitak dlake, povrede, kao i brojna neuhranjena grla (ocena kondicije 1 i 2).
Plesniva silaža i loš kvalitet stočne hrane su takođe utvrđeni. Kao zaključak,
preliminarni rezultati potvrđuju snažnu potrebu za monitoringom farmi, kao i
razgovorima sa upravnicima farmi o neophodnim merama koje se odnose na
dobrobit životinja koje moraju biti preduzete odmah.
References
EFSA (2009): Scientific Report on the effects of farming systems on dairy cow
welfare. European Food Safety Authority, Parma, Italy, 12-16.
GRANDIN T. (2010): Improving Animal Welfare: A Practical Approach,
Cambridge: Cambridge Press, 54-56.
HASKELL M.J., RENNIE L.J., BOWELL V.A., BELL M.J. AND LAWRENCE
A.B. (2006): Housing system, milk production, and zero-grazing effects on
lameness and leg injury in dairy cows. Journal of Dairy Science, 89, 4259–4266.

Study of animal welfare status ...
HRISTOV S., STANKOVIĆ B., TODOROVIĆ JOKSIMOVIĆ M., MEKIĆ C.,
ZLATANOVIĆ Z., OSTOJIĆ-ANDRIĆ D. AND MAKSIMOVIĆ N. (2011a):
Welfare problems in dairy calves. Biotechnology in Animal Husbandry 27, 1417-
1424.
HRISTOV S., ZLATANOVIĆ Z., STANKOVIĆ B., OSTOJIĆ-ANDRIĆ D.,
DAVIDOVIĆ V., JOKSIMOVIĆ TODOROVIĆ M., PLAVŠIĆ B. AND
DOKMANOVIĆ M. (2011b): Procena dobrobiti krava u slobodnom sistemu
držanja [Dairy cows welfare assesment in loose stalls]. Veterinarski glasnik, 65,
399-408.
HRISTOV S., STANKOVIĆ B., ZLATANOVIĆ Z. AND PLAVŠIĆ B. (2011c):
The most significant predisposing factors and causes of lameness of dairy cows.
Proceedings of International Scientific Symposium of Agriculture "Agrosym
Jahorina 2011", 82-90.
HUXLEY J.N, BURKE J., RODERICK S., MAIN D.C.J. AND WHAY H.R.
(2004): Animal welfare assessment benchmarking as a tool for health and welfare
planning in organic dairy herds. Veterinary Records, 155, 237–239.
KOSSAIBAT M.A. AND ESSLEMONT R.J. (1997): The costs of production
diseases in dairy herds in England. Veterinary Journal, 154, 41–51.
LOMBARD J.E., TUCKER C.B., VON KEYSERLINGK M.A.G., KOPRAL C.A.
AND WEARY D.M. (2010): Associations between cow hygiene, hock injuries,
and free stall usage on US dairy farms. Journal of Dairy Science, 93, 4668–4676.
MATTHEWS R. AND WASSMANN R. (2003): Modelling the impacts of climate
change and methane emission reductions on rice production: a review. European
Journal of Agronomy, 19, 573-598.
OSTOJIĆ-ANDRIĆ D., HRISTOV S., NOVAKOVIĆ Ž., PANTELIĆ V.,
PETROVIĆ M.M., ZLATANOVIĆ Z. AND NIKŠIĆ D. (2011): Dairy Cows
Welfare Quality In Loose Vs. Tie Housing System. 3rd International Congress
“New perspectives and Challenges of Sustainable Livestock production” Belgrade,
Republic of Serbia, 5-7th October 2011, Biotechnology in Animal Husbandry, 27,
975-984.
PHILLIPS C.J.C. AND MORRIS I.D. (2002): The ability of cattle to distinguish
between, and their preference for, floors with different levels of friction, and their
avoidance of floors contaminated with excreta. Animal Welfare, 11, 1: 21-29.
POTTERTON S.L., GREEN M.J., HARRIS J., MILLAR K.M., WHAY H.R.
AND HUXLEY J.N. (2011): Risk factors associated with hair loss, ulceration, and
swelling at the hock in freestall-housed UK dairy herds. Journal of Dairy Science,
94, 2952–2963.
RODENBURG J. (2000): Body Condition Scoring of Dairy Cattle. Factsheet, 12,
92-99.
RODRIGUEZ-LAINZ A., HIRD D.W., CARPENTER T.E. AND READ D.H.
(1996): Case-control study of papillomatous digital dermatitis in Southern
California dairy farms. Preventive Veterinary Medicine, 28, 117-131.
515

516
R.T. Gudaj et al.
RUTHERFORD K.M.D., LANGFORD F.M., JACK M.C., SHERWOOD L.,
LAWRENCE A.B. AND HASKELL M.J. (2009): Lameness prevalence and risk
factors in organic and non-organic dairy herds in the United Kingdom. Veterinary
Journal, 180, 95–105.
SCHUKKEN Y.H., GROMMERS F.J., VAN DE GEER D., ERB H.N. AND
BRAND A.(1990): Risk factors for clinical mastitis in herds with a low bulk milk
somatic cell count. 1. Data and risk factors for all cases. Journal of Dairy Science,
73, 3463-71.
SPRECHER D.J., HOSTETLER D.E. AND KANEENE J.B. (1997): A lameness
scoring system that uses posture and gait to predict dairy cattle reproductive
performance. Theriogenology, 47, 1179–1187.
STULL C.L., REED B.A. AND BERRY S.L. (2005): A Comparison of Three
Animal Welfare Assessment Programs on California Dairies. Journal of Dairy
Science, 88, 1595-1600.
TREVISI E., BIONAZ M., PICCIOLI-CAPPELLI F. AND BERTONI G. (2006):
The management of intensive dairy farms can be improved for better welfare and
milk yield. Livestock Science, 103, 231-236
VEISSIER I. AND EVANS A. (2010). Principles and criteria of good Animal
Welfare. Welfare Quality Fact Sheet. Available from http://www.welfarequality.
net/everyone/41858/5/0/22 (accessed Oct 14 2011).
VON KEYSERLINGK M.A.G., RUSHEN J., DE PASSILLÉ A.M. AND WEARY
D.M. (2009): Invited review: the welfare of dairy cattle—Key concepts and the
role of science. Journal of Dairy Science, 92, 4101–4111.
WHAY H.R., MAIN D.C.J., GREEN L.E. AND WEBSTER A.J.F. (2003):
Assessment of the welfare of dairy cattle using animal-based measurements: direct
observations and investigation of farm records. Veterinary Record, 153, 197-202.
Received 14 December 2011; accepted for publication 12 March 2012

Biotechnology in Animal Husbandry 28 (3), p 517-528, 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.08’3
DOI: 10.2298/BAH1203517P
SOME IMPORTANT FACTORS AFFECTING FERTILITY
IN SHEEP
M.P.Petrovic 1 ,V. Caro Petrovic 1 , D. Ruzic Muslic 1 , N. Maksimovic 1 , Z.
Ilic 2 , B. Milosevic 2 , J.Stojkovic 2
1 Institute For Animal Husbandry, Belgrade, Serbia
2 University of Pristina, Faculty of Agriculture, Kopaonicka bb, 38219 Lesak, Serbia
Corresponding author: milanppet@yahoo.com
Review paper
Abstract: Efficiency of sheep production is conditioned by fertility.
According to some authors number of offspring obtained per lambing is more
important than gain of weight. Genetic relationships involving reproductive traits
were seldom studied. Reproductive traits have low heritabilities, a discrete
phenotypic expression, and are expressed only in sexually mature ewes leading to
low selection intensities and long generation intervals. Documentation of realized
selection response is also often complicated by the low heritabilities of fertility
traits. Existence of a major gene affecting prolificacy had been suggested and at
that time there were many sceptics who strongly doubted that a trait as complex as
reproduction could be profoundly influenced by a single gene. Major genes
affecting prolificacy in sheep was founded. A mutation in the bone morphogenetic
protein 15 gene (BMP15, also known as GDF9B) responsible for high prolificacy
in Inverdale sheep had been discovered and evidence of segregating major genes
was being reported from flocks around the world. Development of gene mapping
techniques, and locating alleles that are responsible for the fertility of sheep began
a new chapter in predicting and controlling the fertility of sheep. The beneficial
effects of nutrition on reproduction in sheep are well known. This procedure is
known as flushing. The effect of Body Condition Score (BCS), before mating,
during mating and after mating period, on reproductive efficiency of different
breeds of sheep in the different rearing systems were studied. The farm manager
has the ability to control or at least to manipulate the factors that have an impact on
fertility.
Keywords: sheep, fertility, genetics, nutrition, management
Introduction
Fertility is one of the most important parameters of sheep productivity, the
number of offspring obtained per lambing is a good indicator, and according to

518
M.P.Petrovic et al.
some authors (Petrović, 2000) it is more important than gain of lambs. This means
that biological efficiency of sheep in regard to meat, milk and wool production is
conditioned by fertility ( Notter et al., 2000). Low fertility is a continuing problem
in Australian flocks (Scaramuzzi and Lindsay 1986).
Success in selection of sheep in regard to their fertility greatly depends on
genetic variability of reproductive components (Petrović et al., 1997, 2001, 2002,
2007). Heritability for onset of puberty is low and within the interval of 0,1 to 0,26
(Petrović, 2000). Range or norm of ovulation in sheep is still great challenge for
the science, regardless of the fact that it is generally known that it is under control
of larger number of genes, i.e. its poligeneous character. Activities directed to gene
identification responsible for this trait are in progress (Galloway et al., 2000, Davis
et al., 2001, Grant et al., 2001, Wilson et al., 2001).
For traits of sheep fertility, of great importance is also embryonic
mortality, which is especially present during first four weeks of pregnancy, when
20 - 30% of fertilized egg cells can be lost, and which is in negative correlation
with ovulation rate (Perez et al., 1994).
Feeding of sheep is also one of the most important factors that influence
the fertility of sheep. There are many controversies about it, how best to prepare a
sheep for mating. Flushing is understood as the rapid increase in ovulation rate of
ewes receiving a nutrient supplementation before mating. Branca, et al., 2000.
Lassoued et al. (2004), showed important interactions between genotype and level
of nutrition. In this sense, in highly prolific ewes like D’Man breed, higher levels
of nutrition prior to and during mating were associated with improved reproductive
performance, but in low prolific breeds such as Queue Fine de l'Ouest, neither
ovulation rate nor lambing rate were affected by the dietary treatment. In a recent
work by Fukui et al. (2010) body weight did not significantly affect fertility.
Oestrus in the majority of sheep has a seasonal character, which means that
the sheep exhibit sexual glow during the summer from Jun to September. It is
linked to many as genetic and external factors (Petrović, 2000). Very often it is
associated with climatic and seasonal conditions over which the producer has little
control. On the other hand there are numerous production factors which influence
fertility and which are amenable to management.
In this paper we would like to touch about the following factors that may
affect the fertility of sheep:
• Genetics
• Nutrition
• Management
Genetics and fertility
Number of offspring and growth of lamb, indicated by body weights and
rate of gain at different phases of growth are among the most economically

Some important factors affecting ...
important and easily-measured traits. Knowledge on the particular trait and phase
of the animal’s growth upon which to base selection is therefore of utmost
importance. Genetics/heredity affects the fertility of farm animals in a variety of
ways. Some animals may be genetically infertile. Genetic differences do exist
between different breed and types of sheep. Romanov sheep commonly have
lambing percentages of 250 while Pramenka tend to have lambing percentages of
close to 110. Genetic mutations can occur which can produce infertility. They can
also give rise to abnormal development in embryos so much so that the young fail
to develop properly and cause fetal death or atrophy. These are called lethal
factors.
Genetic relationships involving reproductive traits were seldom studied.
Reproductive traits have low heritabilities, a discrete phenotypic expression, and
are expressed only in sexually mature ewes leading to low selection intensities and
long generation intervals. Documentation of realized selection response is also
often complicated by the low heritabilities of fertility traits. Based on the research
of Al-Shorepy and Notter (1997), selection for fertility in an annual fall lambing
system resulted in increases in estimated breeding value (EBV) for fertility and in
correlated increases in fall litter size. Pedigree estimates of EBV for fertility were
positively associated with future performance, and fertility EBV was significantly
associated with future fertility. These results suggest that performance in fall
lambing can be improved by selection.
Piper and Bindon (1980) reported the possibility that fecundity of the
Booroola Merino as result from the action of a single major gene or closely linked
group of genes affecting ovulation rate. This was the first time that the existence of
a major gene affecting prolificacy had been suggested and at that time there were
many sceptics who strongly doubted that a trait as complex as reproduction could
be profoundly influenced by a single gene. Two decades later, some groups of
researchers (Mulsant et al., 2001., Souza et al., 2001., Wilson et al., 2001),
simultaneously discovered that the inheritance of prolificacy observed in the
Booroola Merinos was the result of a mutation in the bone morphogenetic protein
1B receptor (BMPR- 1B). Meanwhile the notion of major genes affecting
prolificacy in sheep was no longer regarded as genetic heresy. A mutation in the
bone morphogenetic protein 15 gene (BMP15, also known as GDF9B) responsible
for high prolificacy in Inverdale sheep had been discovered by Galloway et al. [16]
and evidence of segregating major genes was being reported from flocks around
the world.
The development of molecular genetics and gene mapping techniques he
began in a period of locating themselves alleles that are responsible for the fertility
of sheep (Table 1). Thus began a new chapter in predicting and controlling the
fertility of sheep.
519

520
M.P.Petrovic et al.
Table 1. Some known and putative major genes for prolificacy in sheep (Davis, 2004)
Gene Name Allele Chr Breed
BMPR-1B Booroola FecBB 6 Merino
BMP15 Inverdale FecXI X Romney
BMP15 Hanna FecXH X Romney
BMP15 Belclare FecXB X Belclare
BMP15 Galway FecXG X Belclare and Cambridge
- Woodlands FecX2W X Coopworth
- Lacaune FecLL 11 Lacaune
Selection for increased fertility of sheep causes more egg cells to be shed
per ovulation and if a ewe normally ovulates a large number of egg cells the
chances will be good that, when conditions are unfavorable. Sheep will then have a
chance to at least one cell can be fertilized and thus resulted in offspring. Use of
physiological quantitative parameters in sheep improvement in increase of fertility
is one of the procedures which can lead to success in sheep selection on fertility
(Petrovic et al., 2007).
Favorable environmental conditions like good feeding and management
have the same influence, as selection, and the results attained in this manner are
faster and more spectacular. Seeing that more multiple births occur under favorable
conditions it is a very good method to identify ewes and rams which have genetic
potential for fertility.
Nutrition and fertility
The beneficial effects of nutrition on reproduction in sheep are well known
(Forcada and Abecia, 2006). In particular, nutrition is one of the main factors
affecting ovulation rate. Generally sheep will be at grass for half of the year,
however during winter or at lambing they may be housed or given extra feed at
grass. It is important to get the nutrition right to prevent loss of body condition or
problems at lambing. Poor nutrition may cause irregular cycles in females, reduced
ovulation, weak offspring, and pregnancy toxemia or reduced twinning. In males
poor nutrition may reduce sperm quantity and quality. Abadjieva et al.,(2011)
informed that the fertility is determined by a multi-hormonal effect, included not
only sex and gonadotropic hormones, but also "metabolic" hormones. A functional
defect in any of the components of this hormonal complex directly affects
reproduction. They also add that new scientific data confirms the strictly
dependence of reproduction on energy sources and metabolic state and shows their
signaling ways. The both have a big importance for good fertility.
The amount of food given to ewes immediately before fertilization is also
of considerable importance. Experiments have shown that, if at that stage, given to

Some important factors affecting ...
ewe a generous plane of nutrition she is likely to shed more eggs than normal. This
results in a higher lambing percentage by increasing the number of twin births.
This procedure is known as flushing. Special attention must be paid to mineral and
vitamin supplement.
Effect of Body Condition Score (BCS), live weight (static effects) and
changes in BCS and live weight (dynamic effects) before mating, during mating
and after mating period, on reproductive efficiency of different breeds of sheep in
the different rearing systems were studied ( Cam et al., 2010, Aliyari et al., 2012).
Most of researchers have reported that, the absolute effects of BCS and live weight
than their variations have greater impact on sheep reproduction efficiency, which
suggest the importance of breed and interactions with nutritional and physiological
conditions and its impact on reproduction efficiency (Gunn, 1983; Koycegiz et al.,
2009). A correlation exists between BCS, live weight and amount of reserving
body fat (Oregui et al., 1997). Also to prediction of adult weight of sheep with
different genotypes, BCS is suitable (Zygoyiannis et al., 1997).
Live weight is a combination of skeleton size and BCS and it’s not a good
representative to evaluate the reproductive efficiency. Adults weight is affected by
factors such as skeleton scheme (body size), fullness or hunger trap (filled or empty
digestive tract) or being wet wool (Demirel et al., 2004). Many researchers have
reported that, fertility affected by BCS (Doney et al., 1982; Guerra et al., 1972;
Gunn, 1983; Koyuncu, 2005; Madani et al., 2009). Garcia et al. (2002) showed the
significant effect of body to initiation of lambs' puberty.
Effect of Body Condition Score, on some reproductive performance of
Afshari ewes was studied by Aliyari et al., 2012 (table 2).
Table 2. Effect of BCS on Mean ±SE reproductive traits (Aliyari et al., 2012)
Trait BSC(2) BSC(2.5) BSC(3) BSC(3.5) ±SE
Lamb born joined ewes 1.24 1.30 1.40 1.05 9.60
Kg lambs born/ joined ewes 6.70 6.71 7.40 5.54 0.45
No of mating conception 1.36 1.20 1.15 1.20 0.09
Birth weight (kg) 5.36 5.15 5.34 5.29 0.10
Weaning weight (kg) 32.50 32.83 33.41 34.87 0.86
Pregnancy duration (day) 151.72 151.31 151.18 151.05 0.15
Ewe conceived at 1 st estrus 78.00 82.00 86.00 86.00 9.60
Kg lambs born/lambing 6.99 7.10 7.56 7.02 0.38
The effect of nutrition on ovulation rate is more evident over a period and
when the nutritional treatments are done in the transitional period between
anoestrus (seasonal or lactational) and the breeding season. Nutrition has a smaller
influence on sexual activity than on ovulation rate. In contrast, the modification of
reproductive seasonality in ewes that have a moderate level of fat reserves using
the level of nutrition as the only management tool seems to be difficult.
521

522
M.P.Petrovic et al.
In the Mediterranean environment, however, it is possible to overcome the
regulating effect of photoperiod on reproductive seasonality in ewes that have a
moderately high level of fat reserves. Results of Forcada and Abecia (2006) show
a consistent reduction (P < 0.05) in the duration of seasonal anoestrus in ewes
maintained from November to September at a constant, moderately high body
condition score (BCS-2.9) when compared to females having a lower body
condition score (BCS-2.3) (Fig. 1).
Figure 1. Oestrous activity and ovulation rate of Rasa Aragonesa ewes
maintained at two constant
levels of body condition score (BCS) throughout the year: moderately high (2.8)
(filled bars and respectively) and moderately low (2.3) (white bars and
respectively) (Forcada et al., 2006)
Signaling mechanisms for the provision of nutritional feedback from
peripheral tissues to the hypothalamus include leptin and insulin, since their

Some important factors affecting ...
secretion is influenced by both body condition (BCS) and food intake in sheep
(Marie et al. 2001). Both hormones are implicated in nutritional modulation of
reproduction (Blache et al. 2000a) and in appetite/bodyweight regulation (Schwartz
et al. 2000).
Body reserves (long-term) and food intake (short-term) both contribute nutritional
feedback to the hypothalamus. Reproductive neuroendocrine output (GnRH/LH) is
stimulated by increased food intake and not by high adiposity in sheep, but it is
unknown whether the appetite regulating hypothalamic neurons show this
differential response.
The amount of food given to the pregnant mother also influences birth
weight of offspring and heavier offspring have a better chance of surviving.
Conversely, the undersized new born animal has less chance of surviving and
frequently dies in the first few days following birth. Such an animal is less able to
maintain its body temperature and may die quickly if born in cold weather. If overfed,
the pregnant animal may suffer difficulties in birth which could also lead to
death of either the offspring or mother or both. Under nutrition in late pregnancy,
particularly in ewes carrying twins may cause pregnancy toxemia. This has given
rise to the practice of steaming up. Steaming up is feeding technique where females
are put on a rising plane of nutrition in the latter stage of pregnancy. It increases
birth weight and milk production.
Management and fertility
The farm manager has the ability to control or at least to manipulate the factors that
have been discussed above. Different ewe farms have different management
practices and this may have an impact on fertility. Examples of how farmers
control or manipulate these factors include:
• Selection of both male and female animals on the basis of their ability to
produce offspring
• Ensuring the correct proportion of male to female animals if using natural
mating
• Selecting animals suited to the environment
• Providing correct nutrition for the stage of production of the animals
• Selecting paddocks that minimize the detrimental effects of environmental
factors for newly born animals
• Use of strategic worm control program
• Use of a suitable vaccination program
523

524
M.P.Petrovic et al.
The significant effect of the farm management has been described in
several studies (Anel, et al., 2005, Paulenz, et al., 2002). Planning of intervals
between lambings, choosing females to inseminate might slightly improve the
fertility results (David et al. (2008). The need for a resting period for the ewe after
lambing to allow uterine involution is well known. However, sometimes the
increasing reproductive rate imposed by the demanding production system involves
short resting periods from lambing to AI, which affects fertility in a negative way.
According to Bodin et al. (1999), reducing the lambing-AI interval to below 40-50
days induces a significant decrease in fertility, even after natural mating. Most
authors recommend not inseminating ewes any sooner than 50 days post-partum
(Anel, et al., 2005). High body temperatures produced in rams by high summer
temperatures is a cause of poor quality semen. High temperatures can also affect
mating with reduced sexual activity. Conception rate is also reduced. This affects
the number of offspring born. Reproductive planning (intervals between lambings,
season, age of first mating, AI technique, etc.) and animal handling (feeding,
health, preparation of AI lots, etc.) have a great effect on fertility results (Anel, et
al., 2005). David et al. (2008), using a joint model combining two main traits, one
relative to female and the other relative to the male, reported that the main
variation factors of AI success were relative to non-sex-specific effects and to
female effect, suggesting that choosing females to inseminate might slightly
improve the AI results.
Conclusion
Based on the results presented by various authors, it could be concluded
that the fertility of sheep is very complex property, and because of its importance
in achieving success in sheep production, it has been studied from various aspects.
Reproductive traits have low heritability, discrete phenotypic expression and are
expressed only in sexually mature ewes, leading to the selection of low intensity
and long generational intervals. The use of certain physiological parameters in the
selection can contribute to the success. Major genes affecting fertility have been
found in many sheep around the world. The development of techniques to map the
location of genes and alleles that are responsible for the fertility of sheep, open a
new chapter in predicting and controlling the fertility of sheep. The beneficial
effects of nutrition on reproduction in sheep are well known. Influence of body
condition score (BCS), before mating, during mating and after mating on the
reproductive efficiency of different breeds of sheep also studied many authors
mentioned here. Farmer through the management has the ability to control, or at
least manipulate factors that influence the fertility, which is also presented in this
review paper. Despite the many factors that affect fertility of sheep, it appeared that

Some important factors affecting ...
the factors of genetics, nutrition and management has major role in influencing the
fertility and the final successful sheep production.
Važniji faktori koji utiču na plodnost kod ovaca
M.P.Petrović, V.Caro Petrović, D. Ružić Muslić, Z. Ilic, B. Milosević,, J.Stojković,
N. Maksimović
Rezime
Efikasnost proizvodnje ovaca je uslovljena plodnošću. Prema nekim
autorima broja potomaka dobijenih po jagnjenju je važnije od prirasta i mase tela.
Genetski uticaji koji uključuju reproduktivne osobine se retko izučavaju. Plodnost
ovaca ima nizak heritabilitiet, diskretnu fenotipsku ekspresiju, a izražena je samo u
seksualno zrelih ovaca, što dovodi do niskog intenziteta selekcije i dugih
generacijskih intervala. Otkriće major gena koji utiču na plodnost je u to vreme
otkrilo mnogo skeptika koji su snažno sumnjali da takav kompleks reprodukcije
može biti duboko pod uticajem jednog gena. Glavne geni utiču na plodnost kod
ovaca i ovnova. Mutacija u koštanoj morfogenetsko proteina 15 gena (BMP15,
takođe poznatom kao GDF9B) otkrivena je kod mnogih ovaca širom sveta, kao
dokaz segregacije major gena. Razvojem tehnika mapiranja gena i lociranja alela
koji su odgovorni za plodnost ovaca, počeolo je novo poglavlje u predviđanju i
kontrolisanju plodnosti ovaca. Povoljni efekti ishrane na reprodukciju u ovaca su
dobro poznati. Ovaj postupak je poznat kao flushing, pred oplodnju. Uticaj
kondicije tela (BCS), pred parenje, tokom parenja i posle parenja na reproduktivnu
efikasnost različitih rasa ovaca u različitim podizanju sistema su takođe proučavali
mnogi ovde pomenuti autori. Farmer putem menadžmenta ima mogućnost da
kontroliše, ili bar da maniupulate faktorima koji imaju uticaj na plodnostovaca, a
što je takođe predstavljeno u ovom preglednom radu.
References
ABADJIEVA D., SHUMKOV K., KISTANOVA E., KACHEVA D., GEORGIEV
B. (2011):
Opportunities for the improvement of the reproductive performances in female
animals .Biotechnology in animal husbandry 27, 365-372.
ALIYARI D., MOEINI M.M., SHAHIR M.H., SIRJANI M.A.(2012): Effect of
Body Condition Score, Live Weight and Age on Reproductive Performance of
Afshari Ewes. Asian Journal of Animal and Veterinary Advances, 7, 904-909.
525

526
M.P.Petrovic et al.
AL-SHOREPY S. A., NOTTER D. R. (1997): Response to Selection for Fertility
in a Fall-Lambing Sheep Flock. J. Anim. Sci. 75:2033–2040.
ANEL L., KAABI M., ABROUG B., ALVAREZ M., ANEL E., BOIXO J. C., DE
LA FUENTE L. F., DE PAZ P.(2005): Factors influencing the success of vaginal
and laparoscopic artificial insemination in Churra ewes: a field assay.
Theriogenology, 63, 4, 1235-1247.
BLACHE D., CHAGAS L.M., BLACKBERRY M.A., VERCOE P.E., MARTIN
G.B.(2000): Metabolic factors affecting the reproductive axis in male sheep.
Journal of Reproduction and Fertility. 120, 1–11.
BODIN L., ELSEN J. M., HANOCQ E., FRANCOIS D., LAJOUS D.,
MANFREDI E., MIALON M. M., BOICHARD D., FOULLEY J. L., SAN-
CRISTOBAL-GAUDY M., TEYSSIER J., THIMONIER J., CHEMINEAU
P.(1999): Genetics of reproduction in ruminants. Productions Animales, 12, 2, 87-
100.
BRANCA A., MOLLE G., SITZIA M., DECANDIA M., LANDAU S.(2000):
Short-term dietary effects on reproductive wastage after induced ovulation and
artificial insemination in primiparous lactating Sarda. Animal Reproduction
Science, 63, 1-2 , 0378-4320.
CAM M.A., OLFAZ AND M., SOYDAN E.(2010): Body measurements reflect
body weights and carcass yields in karayaka sheep. Asian J. Anim. Vet. Adv., 5:
120-127.
DAVIS G.H., DODDS K.G., WHEELER R. AND JAY N.P. (2001):Evidence that
an imprinted gene on the X chromosome increases ovulation rate in sheep Biology
of Reproduction.64 216–221.DAVIS G.H. (2004): Major genes affecting ovulation
rate in sheep. Genet. Sel. Evol. 37, 1, 11–23.
DONEY J.M., GUNN R.G., HORAK F. (1982): Reproduction. In: Sheep and Goat
Production, Coop, I.E. (Ed.). Elsevier Scientific, Amesterdam, pp: 57-80.
FORCADA F., ABECIA J.A. (2006): The effect of nutrition on the seasonality of
reproduction in ewes. Reprod. Nutr. Dev. 46 ,355–365.
FUKUI Y., KOHNO H., OKABE K., KATSUKI S., YOSHIZAWA M., TOGARI
T., WATANABE H, (2010): Factors affecting the fertility of ewes after intrauterine
insemination with frozen-thawed semen during the non-breeding season. Journal of
Reproduction and Development, 56, 4, 460-466.
GALLOWAY S.M., MCNATTY K.P., CAMBRIDGE L.M. (2000): Mutations in
anoocyte-derived growth differentiation factor gene (BMP15) cause increased
ovulation rate and infertility in a dosage-sensitive manner. Nature Genetics. 25,
279-283.
GARCIA M.R., AMSTALDEN M., WILLIAMS S.W., STANKO R.L.
MORRISON AND C.D.( 2002): Serum leptin and its adipose gene expression
during pubertal development, the estrous cycle and different seasons in cattle. J.
Anim. Sci., 80: 2158-2167.

Some important factors affecting ...
GRANT W., MONTGOMERY S., GALLOWAY M., DAVIS G. H., MCNATTY
K P. (2001): Genes controlling ovulation rate in sheep. Reproduction,121, 843–852.
GUERRA J.C., THWAITES C. JEDEY. T.N, (1972): The effects of components
of body weight on reproductive efficiency in the Merino ewe. J. Agric. Sci., 78:
245-249.
GUNN R.G, (1983): The Influence of Nutrition on the Reproductive Performance
of Ewes. In: Sheep Production, Haresign, W. (Ed.). Butterworths, London, pp: 99-
110.
KOYCEGIZ F., EMSEN E., DIAZ C.A.G., KUTLUCA M, (2009): Effects of
lambing season, lamb breed and ewe parity on production traits of fat tailed sheep
and their lambs. J. Anim. Vet. Adv., 8: 195-198.
KOYUNCU M, (2005): Reproductive performance of kivircik ewes on accelerated
lambing management. Pak. J. Biol. Sci., 8: 1499-1502.MADANI T., CHOUIA F.
ABBAS K, (2009): Effect of oestrus synchronisation and body condition on
reproduction of anoestrous ouled djellal ewes. Asian J. Anim. Vet. Adv., 4: 34-40.
MULSANT P., LECERF F., FABRE S., SCHIBLER L., MONGET P.,
LANNELUC I., PISSELET C.,
RIQUET J., MONNIAUX D., CALLEBAUT I., CRIBIU E., THIMONIER J.,
TEYSSIER J., BODIN
L., COGNIE Y., ELSEN J.M, (2001): Mutation in bone morphogenetic protein
receptor-1B is associated with increased ovulation rate in Booroola Merino ewes,
Proc. Natl. Acad. Sci. 98, 5104–5109.
LASSOUED N., REKIK M., MAHOUACHI M., BEN H.M, (2004): The effect of
nutrition prior to and during mating on ovulation rate, reproductive wastage, and
lambing rate in three sheep breeds. Small Ruminant Research, 52, 1-2 , 117-125.
NOTTER, D. R. (2000): Effects of ewe age and season of lambing on prolificacy
in U.S. Targhee, Suffolk, and Polypay sheep. Small Ruminant Res. 38:1-7.
OREGUI L.M., GABINA D., VICENTE M.S., BRAVO M.V. TREACHER T,
(1997): Relationships between body condition score, body weight and internal fat
deposits in Latxa ewes. Anim. Sci., 65: 63-69.
PAULENZ H., ADNOY T., FOSSEN O. H., SODERQUIST L., BERG K.
A,(2002): Effect of deposition site and sperm number on the fertility of sheep
inseminated with liquid semen. Veterinary Record, 150, 10,42 -49.
PETROVIĆ P.M, ŽUJOVIĆ M., D. NEGOVANOVICA, VLAHOVIC M. MEKIĆ
C., D. ALAVANTIC (1997): Status and prospects of sheep breeding in Serbia.
Biotechnology in Animal Husbandry. 3-4, 83-87.
PETROVIĆ P.M (2000): Genetics and Improvement of Sheep
(monograph).Scientific Book, Belgrade, 365 pp.PETROVIĆ P.M, ŽUJOVIĆ M.,
D. NEGOVANOVIC, STRSOGLAVEC, S., RUZIC D. (2001): The importance of
new selection methods in modern system of sheep breeding. Biotechnology in
Animal Husbandry. 17, 159-167
527

528
M.P.Petrovic et al.
PETROVIC, P.M, ŽUJOVIĆ M., D. NEGOVANOVIC, MUSLIĆ RUZIC D.,
STRSOGLAVEC S. (2002): Some aspects of modern organizations selection
procedures in sheep. Contemporary Agriculture.51, 3-4. 155-150.
PETROVIĆ M.P., RUŽIĆ-MUSLIĆ D., ŽUJOVIĆ M., MEKIĆ C,(2007): Genetic
improvement of fertility in sheep by selection according to physiological
parameters. Biotechnology in animal husbandry. 23, 311 – 321.
SCHWARTZ MW, WOODS SC, PORTED JR, SEELEY RJ & BASKIN
DG.(2000): Central nervous system control of food intake. Nature ,404 ,661–671.
SOUZA C.J., MACDOUGALL C., CAMPBELL B.K., MCNEILLY A.S., BAIRD
D.T. (2001): The Booroola (FecB) phenotype is associated with a mutation in the
bone morphogenetic receptor type 1 B (BMPR1B) gene. Journal of
Endocrinology. 169, 1–6.
WILSON T., WU XI-YANG, JUENGEL J.L., ROSS I.K., LUMSDEN J.M.,
LORD E.A., DODDS K.G., WALLING G.A., MCEWAN J.C., O’CONNELL
A.R., MCNATTY K.P., MONTGOMERY G.W. (2001): Highly prolific Booroola
sheep have a mutation in the intracellular kinase domain of bone morphogenetic
protein IB receptor (ALK- 6) that is expressed in both oocytes and granulosa cells,
Biol. Reprod. 64 1225–1235.
ZYGOYIANNIS, D., STAMATARIS C., FRIGGENS N.C., DONEY J.M.
EMMANS G.(1997): Estimation of the mature weight of three breeds of Greek
sheep using condition scoring corrected for the effect of age. J. Anim. Sci., 64,147-153.
Received 27 august 2012; accepted for publication 20 September 2012

Biotechnology in Animal Husbandry 28 (3), p 529-536 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.052’3
DOI: 10.2298/BAH1203529R
THE EFFECT OF THE SYSTEM OF CROSSING ON
FATTENING PARAMETERS OF WEANED LAMBS
D. Ružić-Muslić 1 , M. P. Petrović 1 , M. M. Petrović 1 , 1 Z. Bijelić, 1 V.
Pantelić 1 , P. Perišić 2 , V. Caro-Petrović 1
1 Institute for Animal Husbandry, Belgrade-Zemun, 11080, Srbija
2 Faculty of Agriculture University of Belgrade, Nemanjina 6, 11080 Belgrade, Republic of Serbia
Corresponding author: muslic.ruzic@.gmail.com
Original scientific paper
Abstract: In this paper, the effect of different systems of crossing of sheep:
two-breed (PxW) and three-breed (PxWxIDF) on production results – body weight
and average daily gain of lambs weaned at the age of 60 days and fattened to age of
120 days, was investigated. Also, the effect of crossing on consumption and
conversion of food and nutrients was monitored. Study was carried out on the
Experimental sheep farm of the Institute for Animal Husbandry, Belgrade-Zemun,
and following breeds were used in the crossing: Pirot Pramenka (P), Wurttemberg
(W) and Ile de France (IDF). Pirot Pramenka was used as the maternal basis, while
Wurttemberg rams were used as sires, and as terminal breed, Ile de France rams
were used. Results have shown that the three-breed crosses of F1 generation,
compared to two-breed crosses, obtained higher body gain for 19.23% with lower
food intake for 5.19%, during fattening period lasting from 60th to 120th day of
age. Regarding feed conversion ratio, they used per 1 kg of body gain 29.61% less
hay and 20.82% less concentrate feeds. Positive biological effect exhibited in
three-breed crosses (PxWxIDF) is consequence of individual heterosis and
maternal heterosis.
Key words: crossing, gain, body mass, feed conversion, heterosis
Introduction
Application of crossing in sheep production is aimed at increase of meat
production and it is very important in programs of improvement of sheep breeding.
Crossing of different sheep breeds is focused on use of the superiority of
specialized sheep breeds with good meat production and transmitting of desirable
genes to their progeny (Farid, 1991; Schoeman, 1995; Petrović, 2000, Chrestha
and Heaney, 2004). Success in crossing, in addition to genetic difference between
crossed populations, greatly determined the system of crossing.

530
D. Ružić-Muslić et al.
In the system of crossing which includes only two breeds only the heterosis
of the individual animal is utilized, whereas in three-breed crossing the heterosis of
dam is added. Full utilization (100%) of the heterosis of the individual animal as
well as the dam results in higher gain and body mass, compared to two-breed
crosses. Use of rams of Ill de France breed as terminal breed caused increase of
body mass in their progeny by 12%, compared to results obtained for rams of
original breed (Boujenane et al , 1998).
Petrović et al. (1995), in the study of the effects of various crossing
combinations: Pirot Pramenka x Merinolandschaf, Pirot Pramenka x Ile de France,
Pirot Pramenka x Merinolandschaf x Ile de France and Pirot improved sheep x Ile
de France, in the period to weaning (90 days of age), have determined that threebreed
crosses used by 17.40% less concentrated food per kilogram of gain,
compared to two-breed crosses.
Objective of present study was to compare production performance of
lambs-crosses of F1 generation: Pirot Pramenka x Wurttemberg and Pirot Pramenka
x Wurttemberg x Ile de France, weaned at the age of 60 days and fattened to the
age of 120 days.
Material and methods
Study was carried out on the Experimental sheep farm of the Institute for
Animal Husbandry, Belgrade-Zemun. In the research, 60 lambs – crosses of F1
generation were used: Pirot Pramenka x Wurttemberg (I) and Pirot Pramenka x
Wurttemberg x Ile de France (II) , weaned at the age of 60 days. All animals were
equal in regard to body weight age and sex/gender (gender ratio 1:1). Initial body
weight of lambs at the beginning of trial was around 18.0 kg.
Nutrition of lambs with forage mixtures was ad libitum, whereas the
quantity of hay was limited and equally distributed to groups. The nutritional
value was calculated according to the French system recommended by INRA
(1988) and Obračević (1990). Structure and nutritional value of used mixtures is
presented in Table 1.
Changes of the body weight average daily gain, consumption and
conversion of food and nutrients were checked in 15 day intervals, whereas the
initial and final body weights were measured three times in three consecutive days.
Statistical processing of obtained data was done using the program Stat.Soft, Inc
(2003) STATISTICA (data analysis software system), version 6, by applying
standard mathematical-statistical methods.

The effect of the system of ...
Table 1.Structure and nutritional value of concentrate mixture used in nutrition of weaned
lambs, %
Feeds %
Corn 73
Sunflower meal 23
Lime 2
Salt 1
Premix 1
Dry matter, g kg- 1 (*) 870
NEM,MJ(*) 7.51
UFV(**) 0.99
Total protein,g/kg(*) 142
RUP 43
PDIN/g/animal/day(**) 102
PDIE/g/animal/day(**) 102
Ca,g(*) 8.4
P,g(*) 4.6
RUP- rumen non-degradable protein; PDIN - protein digested in small intestine depending on the
fermenting nitrogen; PDIE - protein digested in small intestine depending on the fermenting organic
matter
**INRA (1988)
*Obračevic (1990)
Results and Discussion
Indicators of the body mass and realized daily gain are presented in Table
2. It is known that the body mass is the most relevant factor influencing the carcass
value (Rodriguez, 2011). It can be observed in the table that lambs obtained in
three-breed crossing, compared to two-breed crosses, had higher body weight at the
end of the fattening by 2.82 kg or 8.76%. By statistical procedure it was
determined that the system of crossing had very significant effect on body weight
(P

532
D. Ružić-Muslić et al.
with Texel and Ile de France, and established that crosses of F1 generation have
realized average daily gain of 0.237 kg. Also, Petrović et al. (1995), by comparing
the systems of crossing, have stated that three-breed crosses (PxWxIDF), compared
to (PxW), to the age of 90 days, have realized by 21.12% higher daily gain. Use of
Ile de France rams in systems of crossing as terminal breed, influenced the increase
of body mass of lambs-crosses by 12% (Boujenane et al., 1998). Positive
biological effect of three-breed crossing is consequence of the heterosis effect. The
essence of this phenomenon is associated with biochemical and physiological
condition of the sheep organism (Bozkov et al., 1980; Andersen and Christensen,
1981; Petrović et al., 1996). It has been established that interracial crossing of
sheep significantly influences the stimulation of biosynthetic processes which is
manifested through: increased mitochondrial activity, more stable metabolic
processes and greater activity of tissue ferments. Experimental study of a group of
Russian authors (Sanikov and Kazanovskij, 1981; Selkin et al., 1995) revealed that
the activity of aspartate-aminotransferase in lambs-crosses was higher in average
by 19.8% and of alanin-aminotransferase by 16.4%. As concluded by above
mentioned authors, this indicates the connection between the level of activity of
ferments and daily gain of crossbred lambs.
Heterosis effect occurs in all systems of crossing, but not in the same
extent. In case of crossing of two sheep breeds, only the heterosis of the individual
animal is exhibited, whereas in three-breed crossing, 100% heterosis of the
individual animal and 100% heterosis of one of the parents are utilized.
Table 2. Production performance of trial lambs
Parameter
Number of animals
Initial body mass, kg
Initial age, days
Final body mass, kg
Final age, days
Average daily gain, g
** (P

The effect of the system of ...
Results of the comparison of three-breed and two-breed crosses, from the aspect of
consumption and utilization of food and nutrients are presented in Tables 3 and 4.
By analysing the data presented in Table 3, it can be concluded that crosses
PxWxIDF consumed slightly less DM (by 5.19%) compared to PxW crosses.
Table 3. Consumption of food and nutrients of the diet
Parameter
Hay, g/day
Concentrate mixture, g/day
DM, g/day
Total protein, g/day
PDIN, g/day
PDIE, g/day
NEM MJ
System of crossing
PxW PxWxIDF
0.271
0.625
0.790
0.127
87.06
82.99
5.84
0.237
0.614
0.749
0.120
83.01
79.46
5.61
In regard to food conversion ratio, it is apparent that three-breed crosses
used by 29.61% less hay and by 20.82% less concentrate mixture compared to twobreed
crosses. Our results are consistent with results of (Rodriguez et al.,2011) who
have stated that crosses of Merino x Asaf have used by 17% less DM compared to
Asaf lambs. Also, Petrović et al. (1995), who have studied the effects of various
crossing combinations: Pirot Pramenka x Merinolandschaf, Pirot Pramenka x Ile
de France, Pirot Pramenka x Merinolandschaf x Ile de France and Pirot improved
sheep x Ile de France, in the period before weaning (90 days of age), concluded
that crosses products of three-breed crossing have utilized by 17.40% less
concentrate mixture per kilogram of gain, compared to two-breed crosses. More
favourable food conversion ratio in three-breed crosses is consequence of positive
heterosis effect.
This is confirmation of the conclusions made by Shrestha et al. (2004)
claiming that crossing represents economically efficient procedure with aim to
utilize individual heterosis and heterosis of the dam.
Table 4. Consumption of food and nutrients per unit of gain
Parameter
Hay, g/day
Concentrate mixture, g/day
DM, g/day
Total protein, g/day
PDIN, g/day
PDIE, g/day
NEM MJ
System of crossing
PxW PxWxIDF
1.79
4.13
5.21
843
575
548
38.58
1.26
3.27
3.99
642
443
424
29.92
533

534
Conclusion
D. Ružić-Muslić et al.
Based on conducted research and obtained results, the following can be
concluded:
• Three-breed crosses, in comparison to two-breed crosses, in the fattening
period from age of 60 to 120 days, have realized higher gain by 19.23%
(P

The effect of the system of ...
Rezultati su pokazali da su trorasni melezi u odnosu na dvorasne, u tovu od
60. do 120. dana uzrasta ostvarili za 19.23% veći prirast i za 5.19% manje
konzumiranje hrane.
U pogledu konverzije hrane, po kg prirasta su utrošili za 29.61% manje sena
i za 20.82% manje koncentrata.
Pozitivan biološki efekat koji se ispoljio kod meleza (PxWxIDF) je
posledica korišćenja heterozisa individue i heterozisa majke.
References
ANDERSEN E., CHRISTENSEN K. (1981): Physiological explanations of
heterosis, 32 nd Annual Meeting of EAAP, 31 August- 5 September, Zagreb
BOZKOV A.I. SERESEVSKA C.M. MAVTINUK H.M. (1980): Sintez
nukleinovih kislot i belkov v pečeni kris raznih linij v zavisi mosti ot vozrasta.
Biohemija, 45,1,113.
BOUJENANE J., BERRADA D., MIHI S., JAMAI M. (1998): Reproductive
performance of ewes and preweaning growth of lambs from three native Moroccan
breeds mated to rams from Moroccan and improved breeds. Small Ruminant
Research, Volume 27, Issue 3, Pages 203-208.
CHRESTHA J.N.B., HEANEY D.P. (2004): Review Canadian, Outaouais and
ridean Arcott breeds of cheep: 2.Crossbreeding registration and subsequent release
to the Canadian sheep. Small Ruminant Research. Volume 55, Issues 1-3, pages 1-13.
FARID A. (1991): Slaughter and carcass characteristics of three fattahled sheep
breeds and their crosses with Corriedale and Targhee rams. Small Ruminant
Research, Volume 5, Issue 3, Pages 255-271.
INRA (1988). Alimentation des bovins, ovins et caprins. Ed. R. Jarriqe,
INRA, Paris, p. 471
LANZA A., ALEO C., LICITRA G., D'AMIGO S., SHIES, L . (1984): Incrocio di
prima generazione in allevamenti di pecore Pinzirite con impiego di arieti Ile de
France , Texel, Merinolandchaf e Barbareschi. Estratto da Tecnica Agricola n. 4-
Anno XXXVI
OBRAČEVIĆ Č. (1990): Tables of nutritive value of livestock feed in the diet of
ruminants, Scientific Book, Beograd.
PETROVIĆ, P.M., NEGOVANOVIĆ D., ŽUJOVIĆ M. (1995): Nove mogućnosti
povećanja proizvodnje jagnjećeg mesa primenom metoda ukrštanja. IV
Međunarodni simpozijum: Novi pravci razvoja stočarstva. Biotehnologija u
stočarstvu, god. 11, 3-6, Beograd
PETROVIĆ P.M., VLAHOVIĆ M., ŽUJOVIĆ M., NEGOVANOVIĆ D., MEKIĆ
C., RUŽIĆ D. (1996): Neki aspekti biohemijske osnove heterozisa kod ovaca.
Biotehnologija u stočarstvu, god. 12, 3-4, pages 67-73, Beograd
PETROVIĆ P.M. (2000): Genetika i oplemenjivanje ovaca. Naučna, Beograd, 365
535

536
D. Ružić-Muslić et al.
RODRIGUEZ A.B., BODAS R., LANDA R., LOPEZ-CAMPOS O.,
MANTECON A.R., GIRALDEZ F.J. (2011): Animal performance, carcass traits
and meat characteristics of Assaf and Merinox Assaf growing lambs. Livestock
Science, volume 138, Issues 1-3, pages 13-19
SANIKOV M.I., KAZANOVSKIJ S.A. (1981): Biohemičeskie osnovi heterozisa
u ovec. Naučnije trudi VASHIL, 210-218
SCHOEMAN S.J., WET D.R., BOTHA M.A., VAN DER MERWE C.A. (1995):
Comparative assessment of biological efficiency of crossbred lambs from two
composite lines and Dorper sheep. Small Ruminant Research, Volume 16, Issue 1,
Pages 61-67
SELKIN I.I., ČIŽOVA L.N., RABOČEV V.K. (1995): Selekcija ovec na
sovemennom etape. Međunarodnij simpozijum. Biotehnologija u stočarstvu, god.
11, 3-6, pages 55-60.
StatSoft, Inc. (data analysis software system), version 6 (2003)
www.statsoft.com.
Received 14 August 2012; accepted for publication 20 September 2012

Biotechnology in Animal Husbandry 28 (3), p 537-544 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.087.7
DOI: 10.2298/BAH1203537I
THE INFLUENCE OF BIOLOGICALLY ACTIVE
SUPPLEMENT "BIORIL"ON PERFORMANCE OF
FATTENING LAMBS
Z. Ilić 1 , J. Stojković 1 , D. Ružić Muslić 2 , V. Caro Petrović 2 ,
M. P. Petrović 2 , R. D. Djoković 3 , V. S. Kurčubić 3 ,
1 University of Pristina, Faculty of Agriculture, Kopaonicka bb, 38219 Lesak, Serbia
2 Institute for Animal Husbandry, P.O.Box. 23, 11081 Zemun, Belgrade, Serbia.
3 Faculty of Agronomy, Cara Dusana 34, 32000 Cacak, Serbia, University of Kragujevac
Corresponding author: ilzoama@open.telekom.rs
Original scientific paper
Abstract: Investigations were carried out in order to determine whether
the addition of bioactive substances "Bioril" on different percentages in
concentrate (0.3% and 0.6%), has an impact on weight gain and feed conversion in
lambs. For the experiment were used lambs of Mis sheep breed, which at the
beginning of the experiment were 50 days of age. The experiment lasted 42 days,
so it is over when all lambs had 92 days of age. The experiment included 60 lambs
(30 male and 30 female) were divided into three groups-20 lambs per group
(control group I and II and III experimental groups ).The results shows that the
greatest gains made lambs of group III. The difference between body weight of
group I and II was 1.62 kg and was statistically significant (P0.05), but lambs of groups III
and II consumed per unit of gain approximately 5g/kg and 10g/kg less concentrate
than lambs of group I. Most hay consumed by lambs of group I. The difference in
hay consumption between groups I and II is 20 g/kg. Lambs in group III consumed
30 g/kg which was less hay than group I and less 10 g/kg than lambs of group II.
The most energy per kg of gain was consumed by lambs of group III, whereas
lambs in II group consumed the least energy.
lambs
Key words: sheep, bioactive supplement, bioril, fattening characteristics,

538
Introduction
Z. Ilić et al.
In majority countries of Europe, lamb meat is the most important product
of the sheep and growth of lambs has special interest for the farmers. Growth of
lambs depends on a number of genetic and environmental factors (Morris et al.,
2000, Petrović, 2000, Petrović et al., 2011, Hansen and Shrestha, 2002, Notter et
al., 2005). It is known that feeding is the most important factor that influences the
expression of genetic potential of lambs (Santos et al., 2002, Ruzic Muslic et al,
2009). In the European Mediterranean countries lambs are traditionally reared with
their dams in indoor conditions until weaning and there after fattened with
concentrate (Safiudo et aI., 1998). The feeding of lambs is an important
concentration of the meal. Energy is the major dietary element that is responsible
for the different utilization of nutrients and thereby the productivity and gain of an
animal (Bellof and Pallauf, 2004, Pittroff et al., 2006, Hosseini et al., 2008, Khalid
et al, 2011). Because of the small volume of digestive organs of lambs on one side
and greater intensity of growth on the other, some unconventional feeds are used,
such as fats of animal origin (Diaz et al., 2002; Samy,2006; Ružić-Muslić et al.,
2007). In addition to traditional nutrients, there are also various bioactive
supplements. Bioactive Supplements "Bioril" is concentrated salt sodium fatty
acids, which comes from sheep wool fat sweat. The fat content in it reaches over
60%, including up to 90% free fatty acids, 6% of free alcohol, 1% of hydrocarbons,
2% aliphatic alcohol and a minor content of methyl esters of fatty acids. "Bioril"
was developed in All-Russian Scientific Research Institute of Sheep and Goat
Breeding-Stavropol, of the Russian Agricultural Academy (Efremov et al., 1992).
Bioril is highly effective and environmentally clean product promotes active
development of the microflora in the rumen, increased enzymatic processes,
increase the total number of VFA and total lipids, while the hydrogenation
processes are enhanced, there is a rapid growth of body weight and wool growth in
sheep (Ermolovna, 1998, Novopashina, 1999).
The objective of this paper was to investigate the effect of “Bioril” on
fattening characteristics of lambs in conditions outside of Russia.
Material and methods
Investigations were carried out on an experimental farm of sheep at the
Institute of Animal Husbandry, Belgrade-Zemun. For the experiment were used
lambs of Mis sheep, which at the beginning of the experiment were 50 days of age.
The experiment lasted 42 days, so it is over when all lambs had 92 days of age The
experiment included 60 lambs (30 male and 30 female) were divided into three
groups-20 lambs per group (I control group and II and III experimental groups).

The influence of biologically ...
The composition of the concentrate and content of "Bioril" in concentrate are
shown in Table 1.
Table 1. Structure of concentrate mix, %
F e e d s G r o u p s
I II III I II III
Corn 66.3 66.0 65.7
Sunflower meal 30 30 30
Bioril 0 0.3 0.6
Chalk 2 2 2
Salt 0.7 0.7 0.7
Premix 1 1 1
Total 100 100 100
In addition to the concentrate, lambs were fed alfalfa hay ad libitum. Water
also was available ad libitum. Samples were analyzed using the conventional
chemical process - WEENDE. Feed refusals were collected and weighed daily.
Lambs were weighed before feeding initially and then at 7 day interval throughout
the experimental period. After completion of 92-days experimental period, final
live weight of each animal was recorded. Statistical analyses
were performed with StatSoft STATISTICA Version 8.
Table 2. Nutritive value of feeds used in trial
Feeds FU NEM/NME, MJ
Undegradable
protein,%
Concentrate I 1.23 7.28 0.38
Concentrate II 1.30 7.39 0.38
Concentrate III 1.34 7.48 0.39
Hay 0.69 4.12 0.31
Milk 0.39 2.34 0.71
Results and discussion
Body development of fattening lambs during the experiment is given in
Table 3.
It can be seen that the lambs at the start of experiments had the same age
and body weight uniformity, which is varied in the range from 18:38 to 18:40 kg.
This is the stage where lambs are adapted for consuming hay and concentrate and
may give an answer on the composition and quality of nutrients. If we look at the
results of the body weight of lambs at the end of the experiment, we can see that
539

540
Z. Ilić et al.
the lowest weight were in lambs of group I, while the lambs of group II and III had
a higher final body weight.
Table 3. Performance of fattening lambs
C r i t e r i o n Groups
I II III
Initial body weight, kg 18.39 18.40 18.38
Initial age, d 50 50 50
Final body weight, kg 31.75 33.37 33.62
Total gain, kg 13.36 14.97 15.24
Average daily gain, g 318 356 362
The difference between body weight of the I and II group was 1.62 kg and
was statistically significant (P

The influence of biologically ...
Table 4. Feed and nutrients conversion per kg of gain, g/kg
Feeds Groups
I II III
Milk 740 670 730
Concentrate mixture 1830 1820 1825
Alfalfa hay 1350 1330 1320
DM 2770 2751 2748
CP 472 469 468
NEM,MJ/kg 20.61 20.47 20.78
In lambs production is very important that lambs consumed less feed per
unit of gain. Only when we compare the gain with feed conversion, we can get a
true picture of the genotype of lambs and the quality of the food consumed. From
Table 4 we saw that lambs of all groups consumed per kg gain more concentrated
than hay. The difference in feed conversion between groups is not significant
(P>0.05). But lambs of groups III and II consumed per unit of gain approximately
5g/kg and 10g/kg less concentrate than lambs of group I. It is clear that lambs in II
and III groups, which were fed with concentrate containing the "Bioril" consumed
less concentrate per kg of gain. It is seen that the difference between experimental
groups is small, only 5g/kg. Ermolovna (1998) stated that inclusion in the diet of
sheep "Bioril" contributes to the development of microorganisms in the rumen: the
number of bacteria increased by 36.9%, 62% of the simplest. Change the
microflora in the rumen contributes to the intensification of enzymatic processes,
leading to an increase in total VFA in the rumen contents of 12%, mainly due to
acetic and valeric acids. Number of total lipids in the rumen contents increased by
34% hydrogenation processes are enhanced essential fatty acids. It could be
concluded that the higher content of bioactive supplement "Bioril" by
microbiological and biochemical effects in the process of digestion, improved
some sensory properties of the concentrate, which makes it more lambs consumed.
Can say that Bioril has a similar effect of probiotics because, as stated Dutta et
al.(2009) positive effects of probiotics on the rumen environment and performance
of ruminants have been intensively studied because they can beneficially modify
microbial activities, fermentative and digestive functions in the rumen. It is further
stated that probiotics can stimulate specific groups of beneficial bacteria in the
rumen, and has provided mechanistic models that can explain their effects on
animal performance. From the table 4, we can see that most hay consumed by
lambs of group I. The difference between groups I and II is 20g/kg. Lambs in
group III consumed 30g/kg less than group I and less 10g/kg than lambs of group
II. The most energy per kg of gain consumed lambs of group III, whereas lambs in
II group consumed the least energy. Other authors also reported that the energy
level in the diet affects the growth of lambs. Seyed (2009) informed that the feed
intake was higher for group fed on low energy diet compared with other treatment
groups. The weight gain of the lambs fed on high-energy diet was increased by
541

542
Z. Ilić et al.
31.60%, while those fed on low energy diet was decreased by 19.87%. The feed
conversion was better in the group fed on high energy compared to other
experimental groups. Ruzic et al.(2009) reported that With increase of diet
concentration by adding 4% of tallow in forage mixture,tendency of reduction of
consumption of dry matter, energy and protein per kg of gain was observed.
Bahtiyarca et al. (2002) showed that the increase in protein and energy content of
diets decreased the voluntary food intake.
Conclusion
Based on research conducted and the obtained results, we can conclude the
following: the experimental groups of lambs that were fed concentrates with
"Bioril" achieved higher daily gain, compared with the control group (P0.05). The difference in feed conversion between groups is also not
significant (P>0.05). Therefore, it is clear that experimental groups of lambs, which
were fed with the concentrate containing the "Bioril" consumed less concentrate
per kg of gain. We can also conclude that most hay consumed by lambs in group I.
The most energy per kg of gain was consumed by lambs of group III, whereas
lambs in group II consumed the least energy.
Acknowledgements
This study is part of the projects TR 31001 "An environmental approach and
implementation of modern biotechnologies as a basis for the improvement of
ruminant breeding technology", and TR 31053 "Modern biotechnology solutions
in the breeding and feeding of cattle sheep and goats for the production of valuable
and safety food" financially supported by the Ministry of Education, Scienceand
Technological development of the Republic of Serbia.
Uticaj bioaktivnog dodatka Biorila na rezultate jagnjadi u
tovu
Z. Ilić, J. Stojković, D. Ružić Muslić, V. Caro Petrović, M. P. Petrović, R.
Djoković, V. S. Kurčubić
Rezime
Istraživanja su sprovedena kako bi se utvrdilo da li dodavanje bioaktivne
supstance "Bioril" u različitim procentima u koncentratu (0,3% i 0,6%), ima uticaj
na prirast i konverziju hrane u jagnjadi. Za eksperiment su korišćena jagnjad od
ovaca rase Mis, koja su na početku eksperimenta bila 50 dana starosti. Eksperiment

The influence of biologically ...
je trajao 42 dana, tako da je završen kada su jagnjad imala 92 dana starosti. U
eksperiment je uključeno 60 jagnjadi (30 muškog i 30 ženskog pola). Jagnjad su
podeljena u tri grupe-20 jagnjadi po grupi (kontrolna grupa I i II i III
eksperimentalne grupe). Rezultati pokazuju da su ostvareni najveći dnevni prirasti
kod jagnjadi II eksperimentalne grupe. Razlika između telesne mase grupe I i II je
1.62 kg i bila je statistički značajna (P0,05), ali
jagnjad grupe III i II troši po jedinici prirasta oko 5g/kg i 10g/kg manje koncentrata
nego jagnjad grupe I. Više sena troše jagnjad iz I grupe. Razlika u potrošnji sena
između grupa I i II je 20g /kg. Najviše energije po kg prirasta troši jagnjad iz III
grupe, dok jagnjad iz II grupe troši najmanje energije.
References
BAHTIYARCA Y., AKTAS A.H., CUFADAR Y. (2002): The effect of diets with
different levels of energy on fattening performance and carcass characteristics of
Konya merino lambs and muttons. Selcuk Univ. Zir. Fak. Derg., 16: 19-25.
BARANOWSKI A., GABRYSZUK M., JOZWIK A., BERNATOWICZ E.,
CHYLINSKI W. (2007): Fattening performance, slaughter indicators and meat
chemical composition in lambs fed the diet supplemented with linseed and mineral
bioplex. Anim. Sci. Papers Rep., 25: 35-44.
BELLOF G., PALLAUF J. (2004): Deposition of protein, fat and energy in lambs
of the breed German Merino Land sheep. Anim. Sci., 78(3): 369-378.
D. RUŽIĆ-MUSLIĆ, M. P. PETROVIĆ, Z. BIJELIĆ (2009): The effect of beef
tallow in lamb nutrition on fattening and carcass characteristics .Biotechnology in
Animal Husbandry 25:431-438.
DIAZ M.T., VELASCO S., CANEQUE V., LAUZURICA S., RUIZ DE
HUIDOBRO F., PEREZ C., GONZALES J., MANZANARES C. (2002): Use of
concentrate or pasture for fattening lambs and its effect on carcass and meat
quality. Small Ruminant Res., 43, 257 268.
DUTTA T. K., KUNDU S. S., KUMAR M. (2009): Potential of direct-fedmicrobials
on lactation performance in ruminants - A critical review. Livest.
Res.Rural Dev., 10: 219-227.
ERMOLOVNA L. S. (1998): Biologically active substances from the animal side
of raw materials - study the mechanisms of formation and action, the production of
forms used in animal husbandry and medicine. Doctoral dissertation. Russian
academy of agricultural sciences, VNIIOK, Stavropol.
HANSEN C., SHRESTHA J. N. B. (2002): Consistency of genetic parameters of
productivity for ewes lambing in February, June, and October under an 8-month
breeding management. Small Rum. Res., 44, 1-8.
543

544
Z. Ilić et al.
HOSSEINI S. M. (2008): Effect of different energy levels of diet on feed
efficiency, growth rate and carcass characteristics of fattening lambs. J. of Anim. &
Vet Adv., 7(12): 1551-1554.
KHALID M. F. L., SHAHZAD M. A., SARWAR M., REHMAN, A. U., SHARIF
M., MUKHTAR N. (2011): Probiotics and lamb performance. African Journal of
Agricultural Research Vol. 6:5198-5203.
MONTOSSI F., LUZARDO S., DE BARBIERI I., SAN JULIÁN R., FRANCO R.,
GUTIÉRREZ D., BRITO G. (2007): Effect of stocking rate and suplementation on
lambs grazing Lotus corniculatus cv. Inia Draco during summer in Uruguay. In:
Proceedings 53rd ICoMST . Ed. Zhou, G. and Zhang, W. China Agricultural
University, Beijing, China. pp 283-284.
MORRIS C. A., HICKEY S. M., CLARKE J. N. (2000): Genetic and
environmental factors affecting lamb survival at birth and through to weaning. NZ.
J. Agric. Res., 43, 515-524.
NOTTER D. R., BORG R. C., KUHEN L. A. (2005): Adjustment of lamb birth
and weaning weights for continuous effects of ewe age. Anim. Sci., 80, 241-249.
NOVOPASHINA S. I. (1999): Growth of fine-wool lambs at different stages of
ewes lambing and feeding their BAS purivetin. Doctoral dissertation. Russian
academy of agricultural sciences, VNIIOK, Stavropol.
PETROVIC P. M. (2000): Genetic and improvement of sheep. Sci. Book,
Belgrade, pp. 365.
PETROVIC P. M., RUZIC MUSLIC D., CARO PETROVIC V., MAKSIMOVIC
N. (2011): Influence of environmental factors on birth weight variability of
indigenous Serbian breeds of sheep. African Journal of Biotechnology. 10: 4673-
4676.
PITTROFF W, D. H. KEISLER, H. D. BLACKBURN (2006): Effects of a highprotein,
low-energy diet in finishing lambs:1. Feed intake, estimated nutrient
uptake, and levels of productivity for ewes lambing in February, June, and October
under an 8-month quality. Small Ruminant Res., 43, 257 268.
PITTROFF, W., KEISLER, D. H., BLACKBURN, H. D. (2006): Effects of a high
protein – low energy diet in finishing lambs 1 Feed intake estimated nutrient
uptake and levels of plasma metabolites and metabolic hormones. Livest. Prod. Sci.
101, 262–277.
RUŽIĆ D., GRUBIĆ G., PETROVIĆ P.M., NEGOVANOVIĆ D., NEŠIĆ Z.,
PERISIC P., ZUJOVIC M. (2007): The effect of the level of non degradable
protein on digestibility of nutritive substances in fattening lambs. Biotechnology in
Animal Husbandry, 23, 5-6, 131-137.
SANTOS-SILVA J., MENDES I. A., BESSA R. J. B. (2002): The effect of
genotype, feeding system and slaughter weight on the quality of light lambs. 1.
Growth, carcass composition and meat quality. Livest. Prod. Sci., 76, 17-25.
STATISTICA (data analysis software system), version 8. www.statsoft.com.
Received 25 June 2012; accepted for publication 18 August 2012

Biotechnology in Animal Husbandry 28 (3), p 545-552 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.087.7
DOI: 10.2298/BAH1203545S
EFFICENCY OF ZEOLITE BASIS PREPARATION IN
FATTENING LAMBS DIET
J. Stojković 1 , Z. Ilić 1 , S. Ćirić 1 , B. Ristanović 1 , M. P. Petrović 2 , V. Caro
Petrović 2 , V. Kurčubić 3
1 University of Pristina, Faculty of Agriculture, Kopaonicka bb, 38219 Lesak, Serbia
2 Institute for Animal Husbandry, P. O. Box 23, 11081 Zemun, Belgrade, Serbia.
3 Faculty of Agronomy, Cara Dusana 34, 32000 Cacak, Serbia, University of Kragujevac
Corresponding author: jovanips@gmail.com
Original scientific paper
Abstract: The paper presents the results of a research on the impact of
products based on natural zeolite on the production results of fattening lambs. The
experiment involved two groups of lambs (the control - C and experimental – E
groups), each consisting of 15 heads, for a period of 90 days. The meal was made
from sheep milk, feed mixtures for fattening lambs and meadow hay. The test
group lambs, unlike the ones from the control group, were given mixtures based on
natural zeolite. Min-a-Zel S mixture (in the form of 25% composite) was fed to
lambs from birth till their 14th day of life, directly into the mouth, once a day
(before the morning feed), in the amount of 10 ml. Min-a-Zel Plus was given to
them from their 15th day of life, together with the feed mixture (0.5%). Feeding
was at will. The average weight of lambs at the end of the experiment, in
accordance with the sequence of treatments (C:E) was 24.40:26.94 kg (P

546
J. Stojković et al.
ammonia, carbon monoxide, pesticides and herbicides), it helps to achieve better
production results and maintain the health and reproductive performance of
domestic animals (Adamovic et al. 2001, Dakovic et al. 2003, Medakovic and Zaric
2005). Stojković (2006), have astablished that the newborn lambs, that were fed the
product based on natural zeolite through colostrum (Min-a-Zel S in the amount of 5
g/l of colostrum), had more than 50 percent higher concentration of
immunoglobulin (IgG) after the period of six hours. Later, 24 and 48 hours after
the birth, the values of IgG also increased, indicating the contribution of these
drugs in strengthening the immune system of lambs in their first days of life.
The aim of this study was to determine the influence of products based on
natural zeolite (Min-a-Zel S and Min-a-Zel Plus) on major production performance
of fattening lambs. Min-a-Zel S is a 25% composite of a processed zeolite
concentrate, containing 90% of clinoptilolite. Min-a-Zel Plus represents a new
generation of mineral mycotoxin adsorbent, obtained by an organic modification of
a zeolite mineral clinoptilolite with a long-chain quaternary amine.
Material and methods
The experiment involved two groups (the control - C and the experimental-
E groups), each consisting of 15 lambs, and it was organized on a Mramor farm,
near Niš. The experiment was realized in January-March 2011, in a period of 90
days. Lambs were fed a meal consisting of sheep's milk, feed mixtures for fattening
lambs and meadow hay. Lambs were reviewed when they were 60 days old. The
experimental group lambs, unlike the control group ones, received the products
based on natural zeolite. Min-a-Zel S mixture was fed to the lambs from birth to
their 14 th day of life, directly in the mouth once a day (before the morning feed) in
the amount of 10 ml. They were given Min-a-Zel Plus from their 15 th day of life,
together with the feed mixture (0.5%). Lamb feeding was at will. Standard
chemical analysis was conducted on the feed samples used (milk, feed mixtures
and hay) at the beginning of experiment (Table 1).
Table 1. Chemical composition of used feedstuffs (calculated to % of DM)
Parameter Concentrate mixture Meadow hay Sheep milk
Dry matter 88.55 89.50 18.10
Proteins 15.10 11.20 6.05
Ash 5.64 7.15 0.66
Fat 2.70 2.05 7.30
Cellulose 9.90 27.36 0.00
NFE 54.90 41.70 4.20
ME, MJ/kg 7.10 0.56 2.50
Ca (g) 0.74 0.70 0.20
P (g) 0.55 0.51 0.14

Efficency of zeolite basis ...
Body weight of lambs was measured at birth and then at 30, 60 and 90
days of the age. Food consumption was monitored daily. Statistical analysis was
done by the Statistica program, Version 6, StatSoft. Inc. (2003).
Results and Discussion
The results of the achieved lamb weight (Table 2) indicate that the lambs
which received Min-a-Zel S and Min-a-Zel Plus were more advanced. At 60 days
old, lamb body weight, in accordance with the sequence of treatments C:E was
17.79:18.91 kg, and the noticeable differences were statistically significant (P

548
J. Stojković et al.
Throughout the experiment (0-90 days), the achieved gain was 229:256
grams, and the established differences of 27 g or 11.79% were in favor of the
experimental group and were statistically highly significant (P

Efficency of zeolite basis ...
utilization of dry matter, protein and energy during the experiment (0-90 day) was,
except for the first month, noticeably better in lambs in the experimental group
(Table 5).
Table 4. Nutrient intake
Nutrient Experimental Control group Experimental Index
period, days
group Control=100
0-30 0.11 0.12 109.09
31-60 0.49 0.51 104.06
Dry matter, kg 61-90 0.80 0.56 93.75
0.90 0.49 0.48 97.96
0-30 39 40 100.56
31-60 90 94 104.40
Protein, g
61-90 137 128 93.43
0-90 91 91 100,00
0-30 1,60 1.62 101.25
31-60 3,99 4.12 103.26
ME, MJ/kg 61-90 6,15 5.79 94.15
0-90 4,09 4.09 100.00
The consumption of dry matter per kilogram of gain in the experimental
group compared to the control group was reduced by 12.21 percent, and the
consumption of protein and energy by 10.58 percent, which confirms that the
mixtures used (Min-a-Zel S and Min-a-Zel Plus) had a positive influence on food
utilization efficiency. In the control group, of lambs 15 to 20 days old, five of the
lambs developed a diarrhea, which occurred again at 60 to 68 days of life. In the
experimental group of lambs there was no noticeable form of diarrhea, which is
likely due to the mixtures being previously tested.
Table 5. Nutrient utilization
Nutrient Experimental Control group Experimental Index
period, days
group Control=100
0-30 0.41 0.43 104.88
31-6 2.51 2.28 90.84
Dry matter, kg 61-90 3.58 2.80 78.21
0-90 2.13 1.87 87.79
0-30 145 145 100.00
31-60 461 420 91.11
Protein, g
61-90 614 478 77.85
0-90 397 369 89.42
0-30 5.95 5.87 98.65
31-60 20.46 18.39 89.88
ME, MJ/kg 61-90 27.58 21.60 78.32
0-90 17.87 15.98 89.42
549

550
Conclusion
J. Stojković et al.
Utilization of zeolite-based products (Min-a-Zel S and Min-a-Zel Plus) had
a positive effect on growth, utilization and utilization efficiency of dry matter,
protein and energy. Average daily gain was higher in the experimental group, by
27 g or 11.79 percent. The consumption of dry matter per kilogram of gain, in the
same group, was reduced by 12.21 percent, and of protein and energy by 10.58
percent. Lower incidence of diarrhea was observed in the experimental group.
Acknowledgements
This study is part of the projects TR 31001 "An environmental approach
and implementation of modern biotechnologies as a basis for the improvement of
ruminant breeding technology", and TR 31053 "Modern biotechnology solutions
in the breeding and feeding of cattle sheep and goats for the production of valuable
and safety food" financially supported by the Ministry of Education, Science and
Technological Development of the Republic of Serbia.
Efikasnost primene preparata na bazi zeolita u obrocima
jagnjadi u tovu
J. Stojković, Z. Ilić, Slavica Ćirić, B. Ristanović, M. P. Petrović, Violeta
Caro Petrović, V. Kurćubić
Rezime
U radu su prikazani rezultati istraživanja o uticaju preparata na bazi
prirodnog zeolita na proizvodne rezultate jagnjadi u tovu. Ogled je izveden na dve
grupe jagnjadi (kontrolna – K i ogledna – O), po 20 jagnjadi u trajanju od 90 dana.
Obrok se sastojao od ovčjeg mleka, krmne smeše za tov jagnjadi i livadskog sena.
Iskorišćavanje preparata na bazi zeolita (Min-a-Zel S i Min-a-Zel Plus) pozitivno je
uticalo na prirast, iskorišćavanje i efikasnost iskorišćavanja suve materije, proteina
i energije. Prosečan dnevni prirast bio je veći kod jagnjadi ogledne grupe za 27 g ili
11,79 posto. Utrošak suve materije za kilogram prirasta u istoj gupi, bio je manji za
12,21 posto a proteina i energije za 10,58 posto. Manja učestalost javljanja proliva
bila je kod jagnjadi ogledne gupe.

References
Efficency of zeolite basis ...
ADAMOVIĆ M., NEŠIĆ S., STOIČEVIĆ S., TOMAŠEVIĆ-ČANOVIĆ
MAGDALENA (2001): The influence of Min-a-Zel Plus modified organic
adsorbent on the quality of corp silage IX Yugoslav Symposium of forage crops.
Belgeade.
ADAMOVIĆ M., TOMAŠEVIĆ-ČANOVIĆ MAGDALENA, MILOŠEVIĆ.
(2002): The contribution of mytotoxin mineral adsorbents to the increase of
production and food quality. Eco-Conference. Novi Sad.
DAKOVIĆ A., TOMAŠEVIĆ ČANOVIĆ M., RATTINGHAUS G., DONDUR V.,
ĐORĐEVIĆ N., ADAMOVIĆ M., GRUBIĆ G., KOLJAJIĆ V. BOČAROV-
STANČIĆ A. (2003): The influence of Min-a-Zel Plus on biochemical,
microbiological, mucotoxicological parameters of alfalfa silage. Journal of
Agriculturale Sciences, vol. 48, br. 2, str. 171-178.
ILIĆ Z., PEŠEV S., SIMEONOVA VALENTINA, MILOŠEVIĆ B., SPASIĆ Z.
(2005): The influence of zeolite type tufozel on productive characteristics of dairy
cows. Biotechnology in animal hunsbandry, VOL 21 (5-6), p 25-30, UDC 636,
ISSN 1450-9156, Belgrade- Zemun.
ILIĆ Z., PEŠEV S., MILENKOVIĆ M., MILOŠEVIĆ B. (2007): IMPACT ON
THE ZEOLITE USAGE IN DIARY COWS NUTRITION TO THEIR HEALTH
CHARACTERISTICS. ISSN 1450-9156. Biotechnology in animal hunsbandry,
VOL 23 (5-6), p, 25-33, Belgrade-Zemun.
JAŠOVIĆ B., STOJKOVIĆ J., MILENKOVIĆ M., MILOŠEVIĆ B., ILIĆ Z.
(2009): Uticaj mineralne smeše sa pufernim dejstvom na proizvone rezultate
jagnjadi u tovu. Agro-knowalage Jurnal, vol. 10, br. 4, 125-128.
MEDAKOVIĆ V., ZARIĆ V. (2005): Adsorption of mycotoxins by
organozeolites. Colloids and Surfaces B Biointerfaces, 46 (1): 20-25.
MAŠIĆ Z., KLJAJIĆ R., BOČAROV-STANČIĆ S., ŠKRINJAR M. (1999):
Mikotoksini u stočnoj hrani kao factor poremećaja zdravlja životinja. 12
Savetovanje veteinara Srbije, Vrnjačka banja, 65-73.
NIKKHAH A., BABAPOOR M., MORADI-SHAHRBABAK (2001): Effect of
clinoptlloite – rich tuff on the performance of Varmini male lambs. Teheran, Iran.
PEŠEV S., ILIĆ Z., SIMONOVA V., MILOŠEVIĆ B., SPASIĆ Z. (2005): The
influence of the zeolite type tufozel on dairy cows reproductive characteristics.
Biotechnology in Animal Husbandry, vol. 21, br. 5-6, str. 19-24
STATISTICA, verzion 6, StotSoft. Inc (2003). www.statsoft.com.
STOJKOVIĆ J., ADAMOVIĆ M., LEMIĆ J., JAŠOVIĆ B. (2005): The effects of
feeds based on natural zeolite on production results for fsttening lambs.
Biotechnology in Animal Husbandry, vol. 21, br. 5-6, dtr. 49-52.
STOJKOVIĆ J. (2006): Mineral substances in animal feed. Monograph. Niš.
551

552
J. Stojković et al.
STOJKOVIĆ M., GRUBIĆ G., ADAMOVIĆ M., MEKIĆ C., ORLOVIĆ JELENA
(1999): The influence of zeolite on the important production results in fattening
lambs. Collection of scientific papers, 5, 489-495.
SHARIATMANDARI F. (2008): The application of zeolite in poultry production.
Worlds Poultry Science Journal, 64, 76-84.
Received 25 June 2012; accepted for publication 10 August 2012

Biotechnology in Animal Husbandry 28 (3), p 553-561, 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 637.07
DOI: 10.2298/BAH1203553J
THE EFFECT OF DIET SELENIUM SUPPLEMENT ON
MEAT QUALITY
M. Joksimović Todorović 1 , V. Davidović 1 , Lj. Sretenović 2
1
University of Belgrade, Faculty of Agriculture, Nemanjina 6, 11080, Belgrade-Zemun, Republic of
Serbia
2
Institute for Animal Husbandry, 11080 Belgrade-Zemun, Republic of Serbia
Corresponding author: miratodo@agrif.bg.ac.rs
Review paper
Abstract: The health of farm animals depends on many factors and they
all indicate that food plays an important role in preserving health, improving
reproductive and productive characteristics and functioning of immune system.
Besides numerous nutritional factors, antioxidants play a special role in the
struggle for survival and good health state. An oxidative stress represents a
condition induced by generating and reacting of reactive-oxygen species (ROS),
and their toxic products, on different metabolic and physiological processes. The
generating of ROS is induced by both endogenous and exogenous factors. In the
case of the organism inability to expel the causative agents of oxidative stress they
can damage cell lipids, proteins or DNA thus endangering the cell functions. Lipid
peroxidation is a process which can lead to degradation of lipids and damage of
cell membrane. Selenoenzyme glutathione peroxidase can protect organism from
peroxidative damages, decrease the level of malondialdehyde protecting in this
way a muscular tissue from oxidation. Recent research has shown that selenium
has an effect of preserving sensory characteristics of meat and its texture, what is
of great importance for consumers. Animal tissue incorporates supplemented
selenium quickly thus it is possible to produce a selenium enriched meat and eggs.
Selenium also shows positive effects on meat quality: it reduces water loss,
influence the stability of colour, prolongs oxidative protection, and therefore the
time of its shelf life.
Key words: selenium, glutathione peroxidase, estimation of meat quality.
Introduction
A main source of selenium for animals are vegetable mixtures in which this
microelement is found in the form of selenocysteine and selenomethionine. The
content of selenium in plants depends on its quantity in the soil where it is present

554
M. Joksimović Todorović et al.
in the form of salts: selenites, selenates and selenides. Some regions are
selenodefficient, so the plants from these regions contain insufficient quantities of
selenium. Because of that the selenium can be added into diet for animals in the
form of supplement – sodium selenite, selenized yeast and selenomethionine
(Sretenović et al., 2007). The organic selenium is more deposited into the muscle
tissue and animal organs than inorganic one (Todorović et al., 1999; Kim and
Mahan, 2001; Joksimović Todorović et al., 2006; Behne et al., 2009; Juniper et al.,
2009). Inorganic form of selenium has a role of prooxidant, but given in great
quantities (< 5 mg/kg diet) can be very toxic (Seko et al., 1989; Mihailović et al.,
1996a,b, 1997; Todorović et al., 1998).
Lipid peroxidation is a natural process which can cause degradation of lipids,
and therefore damage of cell membrane. Malondialdehyde (MDA) is one of
metabolic products of lipid peroxidation which can move lipid oxidation induced
by reactive oxygen species (ROS) in tissues. Free radicals are very unsteady,
reactive and able to damage DNA, proteins, lipids and carbohydrates. Damage of
DNA can potentiate mutation, translator errors, inhibition of protein synthesis,
whilst damaged proteins can lead to changes in transport of ions and enzyme
activity. By the oxidation polyunsaturated fatty acids can change membrane
structure, permeability and activity of membrane enzymes. Damage of biological
molecules and numerous systems endangers the health status and production
abilities in animals. The activity of selenoenzymes, glutathione peroxidase protects
the organism from peroxidative damages, maintaining the content of MDA on low
level. Reducing of the level of malondialdehydes and increased activity of GSH-Px
in tissues are two major indicators of adequate protection of muscle tissue from
oxidation and likely way of prolonging the shelf life of fresh meat (Zhan et al.,
2007).
Water loss and meat colour are important parameters for estimation of meat
quality. Organic selenium (selenized yeast) reduces water loss and can effect the
stability of the colour of pork and chicken meat (Downs et al., 2000).
Concentration of Se and activity of GSH-Px in some organs
and muscle tissue
Physiological roles of selenium are directly associated with the function of
proteins whose content it is being built into via selenocysteine aminoacid. Over 30
selenoproteins are known, but a physiological role is known for only a few of them.
The first discovered enzyme is glutathione peroxidase (GSH-Px) (Rotruck et al.,
1973). Glutathione peroxidase has a role in cell defence against oxidative damages
in animals catalysing reduction of hydrogen peroxides and lipid peroxidation
(Arthur, 2000; Skřivanová et al., 2007). The activity of GSH-Px in animal blood
plasma depends on the level of selenium intake (Hassan, 1987; Todorović, 1990;

The effect of diet selenium ...
Mihailović et al., 1991). Thus the activity of GSH-Px in animal blood plasma is a
reliable indicator of selenium status, but only on optimal and sub-optimal levels.
Depending on the form of selenium, when increasing over necessary levels, the
activity of enzymes shows the effect of plateau, so that a higher levels of Se, lead
to no further increase of activity (Joksimović Todorović and Jokić, 2005;
Joksimović Todorović et al., 2005). Five forms of this enzyme have been defined,
and they are called family: 1. cytoplasmatic – reduces hydrogen peroxide and
peroxides of free fatty acids, 2. gastrointestinal – can be found in the cells of
intestinal epithelium, 3. plasmatic – reacts with hydrogen peroxides, fatty acids
hydroperoxides, phospholipid hydroperoxides, and has an important role in
antioxidative processes in blood plasma, 4. phospholipid hydroperoxide glutathione
peroxidase – reduces phospholipids and cholesterol hydroperoxides and its reduced
activity leads to increased sensibility to oxidative stress and 5. glutathione
peroxidase 6 – found in olfactory epithelium and embryonal tissue. They are
present in almost all cells of organism, but their distibution into tissue can manifest
certain variations. Certain factors: an individual, age, sex, season and cyclic
changes can control the activity of enzymes, while lately there has been some talk
about genetic regulation.
Certain swine organs (kidney, liver, spleen) have fifteen times greater activity
of GSH-Px than muscle tissue or diaphragm (Daun and Åkesson, 2004). However,
in beef treated by the same levels of Se, the activity of this selenoenzyme was three
times lower. The activity of GSH-Px in organs and muscle tissue in pigs reaches
plateau at 0.2 mgSe/kg diet (Lei et al., 1998). This shows that adding of selenium
into animal diet is an important factor of control of the activity of GSH-Px,
however the response can be modulated by specific factors of tissues. The
localization of selenium in each tissue is complex. Total selenium is found in
soluble and insoluble form. Greater activity of GSH-Px in swine than in bovine
organs show that both forms of selenium (soluble and insoluble) are more
deposited in selenoenzyme swine organs. In bulls the concentration of Se in meat is
increasing from 0.107 µg/g to 0. 223 µg/g when 4 mg Se daily is added into diet
for thirty days (Simek et al., 2002). Se-yeast supplementation to the diet of calves
for veal production at a level of 0.5 mg/kg increased Se levels in muscle and
increased GSH-Px activity in muscle and liver (Skřivanová et al., 2007). Chicken
meat shows different content of Se. In chicks inorganic selenium is deposited in
muscle tissue in very little quantites, and because of that a commercial success can
be attained by selenized yeast (Arnold et al., 1973). The effect of adding selenium
into chicks diet is associated mostly with its participation in preserving
antioxidative system of cells.
In gilts fed different levels of organic selenium - 0, 0.3 and 0.6 mg/kg diet, the
highest content of selenium is found in kidneys, followed by liver and heart, while
the lowest one is found in leg and neck musculature, what can be seen in records
shown in Table 1 (Joksimović Todorović et al., 2006). In the liver the concentration
555

556
M. Joksimović Todorović et al.
of selenium was 3-4 times lower, and in heart musculature even up to 6 times lower
than in kidneys. The concentration of selenium in leg and neck musculature was
two, that is, three times higher in gilts fed 0.3 and 0.6 mg Se/kg diet in relation to
control group. These results are in harmony with the results obtained by other
authors (Mahan et al., 1999; Pavlata et al., 2001; Yaroshenko et al., 2004).
Table 1. Effects of different dietary levels of selenium on its concentration in some tissues,
mg/kg
Group
Tissues
Control (0mgSe/kg food) I (0,3 mgSe/kg food) II (0,6 mgSe/kg food)
mg/kg % mg/kg % mg/kg %
Kidney 1.23 100 1.26 102 1.38 112
Liver 0.337 a
Heart 0.180 a
Leg musculature 0.093 a
Neck musculature 0.090 a
100 0.424 ac
100 0.224 ac
100 0.191 ac
100 0.164 ac
Differences between a and b statistically highly significant (P

The effect of diet selenium ...
electrostatic aversion between filaments, the space between them is being reduced
what leads to shrinkage of myofibrils. The integrity of the membrane of muscle
cells is being deteriorated and therefore its semipermeability (Ashgar et al., 1991)
as well. The quantity of water loss depends on the level of lipid peroxidation
(Macit et al., 2003). A slight increase of pH value affects the activity of GSH-Px
and decreases the content of malondialdehyde, and therefore a water loss in the
loins musculature, in the groups of piglets treated with selenomethionine (Zhan et
al., 2007). However, there are some discrepancies among certain authors. Mahan et
al. (1999) point to the fact that organic selenium has no effect on the quality of pig
meat, whilst the inorganic Se has a harmful effect. These discrepancies between
authors regarding the effect of Se on the quality of meat could be a consequence of
supplementation of different levels of Se or ignorance of halothane genetic status in
piglets.
However, research of Zhan et al. (2007) shows that adding 0.3 mgSe/kg diet in
the form of selenomethionine in piglets, stabilizes better a pink colour of meat than
the same quantity of Na-selenite. The colour of meat depends on pH or on the
content of myoglobines (James et al., 2002), and also on the oxidation of
myoglobines into metmyoglobines. Supplementation of Se in diet has no effect on
the content of myoglobines, however it moderately increases pH value,
significantly increases the activity of GSH-Px and reduces the level of MDA in
muscle tissue. Selenomethionine is an efficient "collector" of strong oxidant
peroxinitrites, which produce nitrogen oxides and superoxides. It provides efficient
protection for biomolecules against oxidations and nitrite reactions.
Baowei et al. (2011) confirmed that adding of different levels of selenized
yeast into diets for geese (0.1, 0.3 and 0.5 mg/kg) improved significantly the
quality of breast musculature by the reduction of water loss, increased toughness of
musculature and increase in the content of myoglobines. These authors established
that adding selenoized yeast in the quantity of 0.3 mg/kg diet is an optimal dose for
adequate antioxidative protection in geese. Also, Perić et al. (2009) established that
supplement of selenized yeast into broiler diets led to lower water loss in meat.
Conclusion
In difference to inorganic selenium, organic selenium is deposited more
effectively in tissues. It provides antioxidative protection, preserves the integrity of
cell membranes, reduces water loss in fresh meat during thermal processing and is
a stabilizator of meat colour. This shows that organic selenium provides quality
meat, has an effect on freshness and therefore prolongs shelf life of meat. Chicken
meat enriched with selenium can satisfy 50% needs for selenium in humans by
consuming 100 g daily. Selenium both improves the health of animals and provides
quality meat for consumers.
557

558
Acknowledgment
M. Joksimović Todorović et al.
This work was financed by the Ministry of Education, Science and Technological
Development, Republic of Serbia, project TR 31086: Optimisation of technological
procedure and zootechnical resources on farms with the aim of improving the
sustainability of milk production.
Efekat dodavanja selena u hranu na kvalitet mesa
M. Joksimović Todorović, V. Davidović, Lj. Sretenović
Rezime
Zdravlje životinja zavisi od brojnih faktora, a sve ukazuje da hrana ima važnu
ulogu u očuvanju zdravlja, poboljšanju reproduktivnih i proizvodnih karakteristika
i funkcionisanju imunskog sistema. Pored brojnih hranidbenih faktora,
antioksidansi imaju posebnu ulogu u borbi za opstanak i dobro zdravstveno stanje.
Oksidativni stres je stanje izazvano stvaranjem i reagovanjem reaktiv-oksigen vrsta
(ROS) i njihovih toksičnih produkata na različite metaboličke i fiziološke procese.
Stvaranje ROS izazivaju endogeni i egzogeni faktori. Ukoliko organizam nije u
stanju da otkloni uzročnike oksidativnog stresa oni mogu da oštete ćelijske lipide,
proteine ili DNA, a time da ugroze funkcije ćelija. Lipidna peroksidacija je proces
koji dovodi do degradacije lipida i oštećenja ćelijske membrane. Selenoenzim
glutation peroksidaza štiti organizam od peroksidativnih oštećenja, smanjuje nivo
malondialdehida i na taj način štiti mišićna tkiva od oksidacije. Izučavanja
poslednjih godina ukazuju da selen utiče na očuvanje senzornih karakteristika mesa
i njegove teksture, što je od velikog značaja za konzumente. Životinjsko tkivo brzo
inkorporira dodati selen i tako je moguće proizvesti selenom obogaćeno meso i
jaja. Selen ispoljava pozitivne efekte i na kvalitet mesa: redukuje gubitak tečnosti,
utiče na postojanost boje, produžava oksidativnu zaštitu, a time i vreme njegove
upotrebe.
References
ARNOLD R.L., OLSEN O.E., CARLSON C.W. (1973): Dietary selenium and
arsenic additions and their effects on tissue and egg selenium. Poultry Science, 52,
854-874.
ARTHUR J.R. (2000): The glutathione peroxidases. Cellualar and Molecular Life
Science, 57, 1825-1835).

The effect of diet selenium ...
ASHGAR A., GRAY J.I., BOOREN A.M., GOMAA E.A., ABOUZIED M.M.,
MILLER E.R., BUCKLEY D.J. (1991): Effect of supranutritional dietary vitamin
E levels on subcellular deposition of alpha-tocopherol in the muscle and on pork
quality. J. Sci. Food Agric., 57, 31-41.
BAOWEI W., GUOQING H., QIAOLI W., BIN Y. (2011): Effects of yeast
selenium supplementation on the growth performance, meat quality, immunity, and
antioxidant capacity of goose. Animal Physiology and Animal Nutrition, 95, 4,
440-448.
BEHNE D., ALBER D., KYRIAKOPOULOS A. (2009): Effects of long term
selenium yeast supplementation on selenium status studied in the rat. Journal of
Trace Elements in Medicine and Biology, 23, 258-264.
CHOCT M., NAYLORY A.J., REINKE N. (2004): Selenium supplementation
affects broiler growth performance, meat yield and feather coverage. Brit. Poult.
Sci., 45, 5, 677-683.
COLE D.J.A. (2000): Selenium, the pig and the human diet. In Lyons T.P. and
Cole D.J.A. eds. Concepts in Pig Science 2000. Nottingham University press, UK,
149-158.
DAUN C., ÅKESSON B. (2004): Glutathione peroxidase activity, and content of
total and soluble selenium in five bovine and porcine organs used in meat
production. Meat Science, 66, 801-807.
DOWNS K.M., HESS J.B., BILGILI S.F. (2000): Selenium source effect on
broiler carcass characteristics, meat quality and drip loss. J. Appl. Anim. Res., 18,
61-72.
HASSAN S. (1987): Bioavailability of selenium in feed stuffs as studied in the
chick. Swedish university of Agricultural Science, 1-65.
JAMES B.W., GOODBAND R.D., UNRUH J.A., TOKACH M.D., NELSSEN
J.L., DRITZ S.S., O , QUINN P.R., ANDREWS B.S. (2002): Effects of creatine
monohydrate on finishing pig growth performance, carcass characteristics and meat
quality. Anim. Feed Sci. Tecnol., 96, 135-145.
JOKSIMOVIĆ TODOROVIĆ M., JOKIĆ Ž. (2005): Uticaj visokih nivoa
neorganskog selena na aktivnost glutation peroksidaze (GSH-Px) u krvnoj plazmi
brojlera. Biotechnology in Animal Husbandry, 21, 3-4, 125-131.
JOKSIMOVIĆ TODOROVIĆ M., JOKIĆ Ž., SINOVEC Z. (2005): Uticaj visokih
nivoa organskog selena na aktivnost glutation peroksidaze (GSH-Px) u krvnoj
plazmi brojlera. Veterinarski glasnik, 59, 3-4, 383-390.
JOKSIMOVIĆ TODOROVIĆ M., JOKIĆ Ž., HRISTOV S. (2006): The effect of
different levels of organic selenium on body mass, bodyweight gain, feed
conversion and selenium concentration in some gilts tissues. Acta Veterinaria,
Beograd, 56, 5-6, 489-495.
JUNIPER D.T., PHIPPS R.H., RAMOS-MORALES E., BERTIN G. (2009): Effect
of high dose selenium enriched yeast diets on the distribution of total selenium and
selenium species within lamb tissues. Livestock Science, 122, 63-67.
559

560
M. Joksimović Todorović et al.
KIM Y.Y., MAHAN D.C. (2001): Effects of high dietary of levels of seleniumenriched
yease and sodium selenite on macro and micro mineral metabolism in
grower-finisher swine. Asian-Aust. J. Anim. Sci., 14 (2), 243-249.
LEI X.G., DANN H.M., ROSS D.A., CHENG W.H., COMBS G.F. Jr.,
RONEKER K.R. (1998): Dietary selenium supplementation is required to support
full expression of three selenium-dependent glutathione peroxidases in various
tissues of weanling pigs. Journal of Nutrition, 128, 130-135.
MACIT M., AKSAKAL V., EMSEN E., ESENBUGA N., AKSU M.I. (2003):
Effects of vitamin E supplementation on fattening performance, non-carcass
components and retail cut percentages, and meat quality traits of Awassi lambs.
Meat Science, 64,1-6.
MAHAN D.C., CLONE T.R., RICHERT B. (1999): Effects of dietary levels of
Se- enriched yease and sodium selenite as Se source fed to growing-finishing pigs
on performance, tissue glutathione peroxidase activity, carcass characteristics and
loin quality. J. Anim. Sci., 77, 2172-2179.
MAROUNEK M., DOKOUPILOVÁ A., VOLEK Z., HOZA I. (2009): Quality of
meat and selenium content in tissues of rabbits fed diets supplemented with sodium
selenite, seleniyed yeast and selenized algae. World Rabbit Science, 17, 207-212.
MIEZELIENE A., ALENCIKIENE G., GRUZAUSKAS R., BARSTYS T. (2011):
The effect of dietary selenium supplementation on meat quality of broiler chickens,
Biotechnol. Agron. Soc. Environ., 15(S1), 61-69.
MIHAILOVIĆ M., TODOROVIĆ M., ILIĆ V. (1991): Effects of dietary selenium
of glutatione peroxidase activity and body weight of growing turkeys. Acta
Veterinaria, 23, 75-80.
MIHAILOVIĆ M., TODOROVIĆ M., JOVANOVIĆ M., PALIĆ T.,
JOVANOVIĆ J., PEŠUT O., KOSANOVIĆ M. (1996a): Toxicity of organic and
inorganic selenium to chicken. 9th International Symposium of Trace Elements in
Man and Animals (Tema 9) - Banff Alberta - Canada 1996. Trace Elements in Man
and Animals, 9, 554-555.
MIHAILOVIĆ M., TODOROVIĆ MIRJANA JOVANOVIĆ M., PALIĆ T.,
JOVANOVIĆ J., PEŠUT O., KOSANOVIĆ M. (1996b): Toxicity of sodium
selenite and selenized yeast to chicken -The Sixt Intrnat. Symp. on selenium in
Biol. and Med. - Peking NR Kina 1996. Program and Abstract, 125.
MIHAILOVIĆ M., TODOROVIĆ M., JOVANOVIĆ M., PALIĆ T.,
JOVANOVIĆ J., PEŠUT O. (1997): Toxicity of selenium to broilers. XI th
International Congres of the World Poultry Association 18-22 August, Budapest,
Hungary.
PAVLATA L., ILLEK J., PECHOVA A. (2001): Blood and tissue selenium
concentrations in calves treated with inorganic or organic selenium compounds – a
comparison. Acta Vet. Brno, 70, 19-26.
PERIĆ L., MILOŠEVIĆ N., ŽIKIĆ D., KANAČKI Z., DŽINIĆ N., NOLLET L.,
SPRING P. (2009): Effect of selenium sources on performance and meat

The effect of diet selenium ...
characteristics of broiler chickens. Journal of Applied Poultry Research, 18, 3, 403-
409.
ROTRUCK J.T., POPE A.L., GANTHER H.E., SWASON A.B., HAFEMAN
D.G., HEEKSTRA W.G. (1973): Selenium biochemical role as component of
glutathione peroxidase. Science, 179, 1957-1963.
SEKO Y., SAITO Y., KITAHARA J., IMURA N. (1989): Active oxygen
generation by the reaction of selenite with reduced glutathione in vitro. In: A.
Wendel (Ed.) Selenium in Biology and Medicine, 33-70, Springer-Verlag, Berlin.
SIMEK J., CHLADEK G., KOUTNIK V., STEINHAUSER L. (2002): Selenium
content of beef and its effect on drip and fluid losses. Animal Science Papers and
Reports 20 (Suppl. 1), 49-53.
SKŔIVANOVÁ E., MAROUNEK M., DE SMET S., RAES K. (2007): Influence
of dietary selenium and vitamin E on quality of veal. Meat Science, 76, 495-500.
SRETENOVIĆ LJ., ALEKSIĆ S., PETROVIĆ P.M., MIŠČEVIĆ B. (2007):
Nutritional factors influecing improvement of milk and meat quality as well as
productive and reproductive parameters of cattle. Biotechnology in Animal
Husbandry, 23, 5-6, book 1, 217-226.
SRETENOVIĆ LJ., NOVAKOVIĆ Ž., PETROVIĆ M.M., TODOROVIĆ M.,
PANTELIĆ V., ALEKSIĆ S., PETRIČEVIĆ M. (2012): Producing of beef meat
enriched with organically bound selenium. Biotechnology in Animal Husbandry,
28, 2, 219-229.
TODOROVIĆ M.(1990): Uticaj deficita selena na aktivnost glutation peroksidaze,
alkalne fosfataze i prirast ćurića u tovu. Magistarski rad, Beograd.
TODOROVIĆ M., MIHAILOVIĆ M. (1998): Toksičnost neorganskog selena u
živine. Savremena poljoprivreda, 1-2, 239-241.
TODOROVIĆ M., MIHAILOVIĆ M., HRISTOV S. (1999): Visoki nivoi selena u
ishrani pilića i njegovo deponovanje u mišićima. Savremena poljoprivreda, 1-2,
257-260.
YAROSHENKO F.O., SURAI P.F., YAROSHENKO Y.F., KARADAS F., NICK
H.C. SPARKS (2004).: Theoretical background and commercial application of
production of Se-enriched chicken. Presented at: XXII World Poultry Congress,
June 8-13, Istanbul, Turkey.
ZHAN X.A., WANG M., ZHAO R.Q., LI W.F., XU Z.R. (2007): Effects of
different selenium source on selenium distribution, loin quality and antioxidant
status in finishing pigs. Animal Feed Science and Technology, 132, 202-211.
Received 14 May 2012; accepted for publication 10 July 2012
561

Biotechnology in Animal Husbandry 28 (3), p 563-573 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 637.04
DOI: 10.2298/BAH1203563T
FATTY ACID PROFILE IN RAINBOW TROUT
(ONCORHYNCHUS MYKISS) AS INFLUENCED BY DIET
D. Trbović, D. Vranić, J. Djinovic-Stojanović, V. Matekalo-Sverak, V.
Djordjević, D. Spirić, J. Babić, R. Petronijević, A. Spirić
Institute of Meat Hygiene and Technology, Kacanskog 13, 11000 Belgrade, Serbia
Corresponding author: aurelija@inmesbgd.com
Original scientific paper
Abstract: With the aim of reinforcement S&T capacities in aquaculture,
studies on the influence of three commercial pelleted diets on fatty acid profile in
rainbow trout production were undertaken. Commercial diets for rainbow trout
contained significantly different quantities of saturated fatty acids (SFA), (p

564
D. Trbović et al.
world (Williams, 1998). Nutritional and health benefits achieved by consumption
of fish increased the demand of these products on the market (Burger and
Gochfeld, 2009). Proteins of high biological value, low fat content and relatively
low cholesterol content, as well as valuable quantities of essential fatty acids makes
fish one of the most appreciable food stuffs in human nutrition (Conor, 2000;
Sidhu, 2003).
Nutritional value of different fish species from aquaculture might be
considered as valuable as nutritional value of fish species from open waters
(Weaver et al., 2008). However, some fish species from inland aquaculture are
more capable than marine fish to desaturate and elongate C18 polyunsaturated fatty
acids to valuable highly unsaturated fatty acids, EPA (eicosapentaenoic acid) and
DHA (docasahexaenoic acid), even when their feed is less rich in these fatty acids
(Tocher et al., 2004).
It has been proved that nutritional value of fish can vary due to species,
diet, location, environmental conditions, age, etc (Petterson et al., 2009; Sicuro et
al., 2010). In farmed trout, where environmental conditions are more constant
throughout the year, development and fish growth as well as fatty acid profile is
more affected by food supply (Almeida et al., 2011). Many literature data indicate
to a strong connection between fish diet and fatty acid profile of fish. (Caballero et
al., 2002; Valente et al., 2007) Under the same rearing conditions, feed reach in n-
3 PUFAs significantly increase the n-3/n-6 ratio in fish (Skalli and Robin, 2004).
Rainbow trout is a salmonid fish species originating from the tributaries of
the Pacific Ocean in Asia and North America. This fish species has been
introduced for food in many countries from almost all continents. Aquaculture of
salmonid fish species started to grow exponentially in 1950s, particularly in
Europe. In Norway, the ocean cage production of salmon trout to supply export
markets started to expand, while inland production of rainbow trout has increased
in many European countries to support domestic markets.
Aquaculture as a country economy in Serbia is mainly related to carp
(Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) production (Trbović et
al., 2009; Vranić et al., 2011). Rainbow trout production presents close to 15% of
the national aquaculture and has been developed on mountain areas, abundant in
cold spring waters. However, rainbow trout production still remains on low
technical and technological level due to lack of investment in aquaculture. With the
aim of reinforcement S&T capacities in aquaculture, studies on the influence of
commercial pelleted diets on fatty acid profile in rainbow trout production were
undertaken.
Materials and Methods
The intensive fish farm
Samples of rainbow trout (Oncorhynchus mykiss) were collected from an
intensive fish farm located 931 m above sea level, on the mountains area of

Fatty acid profile in ...
Zlatibor, Serbia. Water of potable quality is captured directly from the spring and,
by gravity force, through channel system, is directed to the farm, with a capacity of
500 l/s. The annual level of production is about 450 t of fish.
Animals and samplings
Rainbow trout was manually fed commercial extruded sinking pelleted
diets, according to good aquaculture practice. Based on specifications, Diet I
consisted of fish meal (17%), blood meal (13%), fish oil (10%) and other
ingredients, like oil seed cakes and soybean, rapeseed, and sunflower meals. Diet II
consisted of fish meal (27%), soybean products and other ingredients like chops
from soybean extraction, extruded soybean seed, soybean protein concentrate,
wheat, rapeseed chops, and oils (rapeseed, fish and palm oil). Diet III consisted of
fish meal, soybean protein products, corn, wheat, minerals and vitamins.
Marketable size fish, 30-35 cm length and 250- 300 g weight, were
sampled. During each sampling, six fishes were collected along with the
appropriate feed (n=6). Fish fillets, obtained after evisceration and previous
deprivation of skin, tail, head, fins and bones were homogenized in a laboratory
blender (Braun CombiMax 600), separately placed in plastic bags and stored at -
25°C until analyzed. A day before analysis samples were defrosted overnight, at
+4°C.
Chemicals and standards
Analytical-grade solvents were obtained from Merck (Darmstadt,
Germany) and Sigma Aldrich (Germany). Reagent for derivatization of fatty acids
(TMSH 0.25 M in methanol) was purchased from Fluka. Fatty acid standards were
supplied by Supelco (Bellefonte, USA), (Supelco 37 comp. FAME mix 10 mg mL -1
in CH2Cl2).
More detailed data on the extraction of lipids by accelerated solvent
extraction (ASE) and on fatty acids capillary gas chromatography determination
are presented in our previous publications (Spirić et al., 2009; Spirić et al., 2010).
A brief review of accelerated solvent extraction of lipids and fatty acids
determination is presented below.
Accelerated solvent extraction (ASE) of lipids
Total lipids for FA determination were extracted from fish muscle by ASE
(ASE 200, Dionex, Sunnyvale, CA). Extraction was performed in 33 mL stainless
steel extraction cells filled with diatomaceous earth, at 100ºC, under 10.3 MPa
nitrogen pressure, with a mixture of n-hexane and iso-propanol (60:40 v/v), in two
static cycles. Solvent was removed under stream of nitrogen (Dionex Solvent
evaporator 500), at 50°C.
Determination of fatty acids by GC/FID
565

566
D. Trbović et al.
After ASE extraction of total lipids and their transesterification by
trimethylsulfonium hydroxide (EN ISO 5509:2000), fatty acids have been
determined, as methyl esters, by Shimadzu 2010 capillary gas chromatograph
equipped with flame ionization detector (GC/FID) and cianopropyl-aryl HP-88
capillary column. Chromatographic peaks in the extracts were identified by
comparing relative retention times of FAME peaks with peaks in the Supelco 37
Component FAMEs mix standard. Quantification was performed by using
heneicosanoic acid methyl ester as the internal standard. The total share of SFA,
MUFA and PUFA consisted of weight percentages of the appropriate individual
fatty acids (Pavlovski et al., 2011).
Statistical analysis
For data analysis UnscramblerX statistical software (CamoSoft, Norway)
was used. Means within each group were compared with ANOVA and Tukey -
Kramer’s multiple range tests.
Results and Discussion
Data obtained for fatty acid composition (% of total fatty acids) of rainbow
trout diets are presented in Table 1.
The obtained data indicate that the predominant fatty acids in all fish diets
were palmitic (C16:0), palmitoleic (C16:1), oleic (C18:1cis-9), linoleic (C18:2 n-
6), EPA and DHA fatty acid. Commercial diets for trout contained significantly
different quantities of SFAs, p

Fatty acid profile in ...
Table 1. Fatty acid composition (% of total fatty acids) of marketable size trout diets (mean ±
standard deviation)
Fatty acids Diet I Diet II Diet III
14:0 4.29±0.01 B
4.35±0.05 B
6.45±0.02 A
15:0 0.36±0.03 AB
0.33±0.01 B
0.41±0.02 A
16:0 13.10±0.04 C
28.15±0.20 A
19.18±0.02 B
16:1 4.76±0.02 B
4.79±0.02 B
7.33±0.02 A
17:0 0.28±0.03 A
0.28±0.01 A
0.30±0.01 A
17:1 0.77±0.02 B
0.61±0.03 C
1.00±0.01 A
18:0 2.64±0.02 C
5.00±0.04 A
3.84±0.36 B
18:1cis-9 34.82±0.05 A
19.40±0.16 B
18.20±0.13 C
18:1cis-11 3.14±0.02 B
3.35±0.10 B
3.96±0.04 A
18:2 n-6 12.21±0.12 A
12.02±0.37 A
8.06±0.06 B
18:3 n-6 0.06±0.01 C
0.12±0.01 B
0.14±0.01 A
18:3 n-3 5.12±0.06 A
1.33±0.01 C
1.89±0.01 B
20:0 0.35±0.01 B
0.40±0.01 A
0.19±0.01 C
20:1 1.46±0.03 B
1.14±0.01 C
2.38±0.04 A
20:2 0.16±0.04 B
0.19±0.06 B
0.41±0.02 A
20:3 n-6 0.54±0.04 A
0.42±0.20 A
0.47±0.12 A
20:3 n-3 1.35±0.01 B
1.00±0.05 B
2.51±0.20 A
22:1+20:4 0.62±0.01 B
1.24±0.18 A
1.04±0.02 AB
20:5 n-3 8.69±0.02 B
9.11±0.26 B
11.36±0.45 A
22:5 n-3 1.11±0.02 C
1.24±0.04 B
2.39±0.01 A
22:6 n-3 3.90±0.02 C
5.30±0.25 B
8.18±0.41 A
24:1 0.27±0.02 B 0.29±0.01 B 0.35±0.02 A
SFA 21.02±0.11 C
38.50±0.19 A
30.35±0.35 B
MUFA 45.21±0.04 A
29.56±0.19 C
33.21±0.12 B
PUFA 33.73±0.18 B
31.95±0.01 C
36.43±0.46 A
n-3 20.16±0.12 B
17.97±0.45 C
26.33±0.69 A
n-6 13.58±0.07 A
13.98±0.44 A
10.10±0.23 B
n-3/n-6 1.49±0.01 B
1.29±0.07 B
2.61±0.13 A
A, B, C : values in the same row with different superscript are significantly different (p

568
D. Trbović et al.
Table 2. Fatty acid composition of fish lipids (% of total fatty acids) in marketable size trout
(mean ± standard deviation)
Fatty acids Pond I Pond II Pond III
14:0 3.33±0.26 B
15:0 0.15±0.02 C
16:0 15.61±0.83 C
16:1 4.83±0.27 B
17:0 0.20±0.02 B
17:1 0.53±0.05 AB
18:0 3.23±0.22 B
18:1cis-9 31.89±1.65 A
18:1cis-11 3.18±0.08 B
18:2 n-6 10.97±0.61 A
18:3 n-6 0.10±0.02 B
18:3 n-3 2.98±0.30 A
20:1 2.74±0.28 A
20:2 0.61±0.05 A
20:3 n-6 0.54±0.04 A
20:3 n-3 2.14±0.24 A
22:1+20:4 0.99±0.07 A
20:5 n-3 3.73±0.57 A
22:5 n-3 1.87±0.31 A
22:6 n-3 10.39±1.71 A
24:1 0.33±0.03 A
SFA 22.17±1.11 C
MUFA 43.50±2.10 A
PUFA 34.33±2.66 A
n-3 21.12±2.52 A
n-6
n-3/n-6
13.21±0.65 A
1.60±0.20 A
2.82±0.64 B
0.26±0.05 B
20.39±2.62 B
3.72±1.16 B
0.22±0.03 B
0.35±0.16 B
4.91±1.29 A
29.13±4.18 A
4.15±0.48 A
10.09±1.34 A
0.10±0.02 B
2.00±0.39 B
2.68±0.20 A
0.52±0.09 AB
0.36±0.10 B
2.23±0.23 A
1.22±0.22 A
2.95±0.64 A
1.79±0.70 A
9.78±2.15 AB
0.33±0.07 A
28.63±3.47 B
40.36±2.59 A
31.04±4.86 AB
18.75±3.47 AB
12.29±1.08 A
1.52±0.23 A
4.63±0.45 A
0.38±0.01 A
24.65±2.89 A
5.94±0.60 A
0.30±0.02 A
0.60±0.13 A
5.67±1.12 A
21.90±2.00 B
4.45±0.26 A
10.50±0.86 A
0.16±0.02 A
1.56±0.27 C
1.71±0.32 B
0.42±0.04 B
0.28±0.02 B
2.21±0.37 A
1.10±0.22 A
4.26±0.71 A
1.52±0.36 A
7.53±0.41 B
0.39±0.07 A
35.63±3.61 A
35.00±2.89 B
29.38±1.44 B
17.08±1.32 B
12.30±0.85 A
1.39±0.14 A
A, B, C : values in the same row with different superscript are significantly different (p

Fatty acid profile in ...
The highest content of palmitic acid was determined in trout from Pond III
(24.65%) and the lowest in fish from Pond I (15.61%). The content of oleic acid
was the highest in fish from Pond I (31.89%) and the lowest in fish from Pond III
(21.90%). Significantly lower proportion of linoleic acid in Diet III (p0.05), (2.95-4.26%).
The highest content of DHA was established in fish fed Diet I (10.39%) and the
lowest in fish fed Diet III (7.53%). Generally, the content of DHA in trout was
higher than in diets, while EPA content was lower, what is in accordance with data
obtained by Kalyoncu et al. (2010), (EPA: 3.11-5.52%; DHA: 6.98-17.57%). Data
we obtained reflects the fact that fish selectively deposit DHA in the body, as
established by Henderson (1996) and Sargent et al. (2002).
The amounts of SFAs in trout were significantly different and ranged from
22.17 (Pond I) to 35.63%, (Pond III). Quantities of MUFA were in the range from
35.00 (Pond III) to 43.50% (Pond I). The proportions of PUFA ranged from 29.38
(Pond III) to 34.33%, (Pond I). Significant differences (p

570
Conclusions
D. Trbović et al.
Significant differences (p

Fatty acid profile in ...
respektivno. Odnos n-3/n-6 bio je u opsegu 1,29- 2,61. Od ZMK (ukupno ZMK:
22,17-35,63%), u uzorcima ribe, najzastupljenija je bila palmitinska kiselina
(C16:0), (15,61-24,65%). Oleinska kiselina (C18:1 cis-9) je bila najzastupljenija u
grupi MNZMK (21,90-31,89%), (ukupno MNZMK: 35,00-43,50%). Od
polinezasićenih masnih kiselina (ukupno PNZMK: 29,38-34,33%), linolenska
kiselina (C18:2 n-6), (10,09-10,97%), EPA (C20:5 n-3), (2,95-4,26%) i DHA
(C22:6 n-3), (7,53-10,39%) su bile prisutne u značajnim količinama u pastrmci.
Količine n-3 masnih kiselina bile su u opsegu 17,08 do 21,12%, a značajne razlike
(p0,05).
Odnos n-3 i n-6 bio je u opsegu 1,39 do 1,60 i, takođe, nisu ustanovljene značajne
razlike (p>0,05). Dobijeni rezultati ukazuju da se sastav masnih kiselina hrane
odražava na sastav masnih kiselina ribe, sa varijacijama koje ukazuju da je
ugradnja masnih kiselina u tkivu ribe uslovljena određenim metaboličkim efektima.
References
ALMEIDA I., MARTINS H.M., SANTOS S., FREITAS S.G., BERNARDO F.
(2011): Mycobiota in feed for farmed sea bass (dicentrarchus labrax).
Biotechnology in Animal Husbandry, 27, 93-100.
BELL M.V., DICK J.R. (2004): Changes in capacity to synthesise 22:6n-3 during
early development in rainbow trout (Oncorhynchus mykiss). Aquaculture, 235,
393-409.
BURGER J., GOCHFELD M. (2009): Perceptions of the risks and benefits of fish
consumption: Individual choices to reduce risk and increase health benefits.
Environmental Research, 109, 343–349.
CABALLERO M.J., OBACH A., ROSENLUND G., MONTERO D., GISVOLD
M., IZQUIERDO M.S. (2002): Impact of different dietary lipid sources on growth,
lipid digestibility, tissue fatty acid composition and histology of rainbow trout,
Oncorhynchus mykiss. Aquaculture 214, 253-271.
CONOR W.E. (2000): Importance of n-3 fatty acids in health and disease.
American J Clinical Nutrition, 71,171S-175S.
FONSECA-MADRIGAL J., KARALAZOS V., CAMPBELL P.J., BELL J.G.,
TOCHER D.R. (2005): Influence of dietary palm oil on growth, tissue fatty acid
compositions, and fatty acid metabolism in liver and intestine in rainbow trout
(Oncorhynchus mykiss). Aquaculture Nutrition, 11, 241-250.
HARDY R.W., SCOTT T.M., HARRELL L.W. (1987): Replacement of herring oil
with menhaden oil, soybean oil, or tallow in the diets of Atlantic salmon raised in
marine net-pens. Aquaculture, 65, 267-277.
HENDERSON R.J., TOCHER D.R. (1987): The lipid composition and
biochemistry of freshwater fish. Progress in Lipid Research, 26, 281-347.
571

572
D. Trbović et al.
HENDERSON R.J. (1996): Fatty acid metabolism in freshwater fish with
particular reference to polyunsaturated fatty acids. Archives of Animal Nutrition
49, 5-22.
KALYONCU L., YAMAN Z., AKTUMSEK A. (2010): Determination of the
seasoanl changes on total fatty acid composition of rainbow trout, Oncorhynchus
mykiss in Ivriz Dam Lake, Turkey. African Journal of Biotechnology, 9, 4783-
4787.
PAVLOVSKI Z., ŠKRBIĆ Z., LUKIĆ M., LILIĆ S., KRNJAJA V., STANIŠIĆ N.
PETRIČEVIĆ V. (2011): Comparative analysis of fatty acid profile and cholesterol
content in table eggs from different genotype hens. Biotechnology in Animal
Husbandry, 27, 93-100.
PETTERSSON A., JOHNSSON L., BRÄNNÄS E., PICKOVA J. (2009): Effects
of rapeseed oil replacement in fish feed on lipid composition and self-selection by
rainbow trout (Oncorhynchus mykiss). Aquaculture Nutrition, 15, 577-586.
SARGENT J.R., TOCHER D.R., BELL J.G. (2002): The lipids. Pg. 182-257 in
Halver J.E., Hard R.W., ed. Fish nutrition. Academic Press, London, UK.
SICURO B., BARBERA S., DAPRA F., GAI F., GASCO L., PAGLIALONGA
G., PALMEGIANO G.B., VILELLA S. (2010): The olive oil by-product in
“rainbow trout Onchorynchus mykyss (Walbaum)” farming: productive results and
quality of the product. Aquaculture Research, 41, e475-e486.
SIDHU K.S. (2003): Health benefits and potential risks related to consumption of
fish or fish oil. Regulatory Toxicology and Pharmacology, 38, 336–344.
SKALLI A., ROBIN J. H. (2004): Requirement of n-3 long chain polyunsaturated
fatty acids for European sea bass (Dicentrarchus labrax) juveniles: growth and fatty
acid composition. Aquaculture, 240, 399–415.
SPIRIĆ A., TRBOVIĆ D., VRANIĆ D., DJINOVIĆ J., PETRONIJEVIĆ R.,
MATEKALO-SVERAK V. (2010): Statistical evaluation of fatty acid profile and
cholesterol content in fish (Common carp) lipids obtained by different sample
preparation procedures. Analytica Chimica Acta, 672, 66-71.
SPIRIĆ A., TRBOVIĆ D., VRANIĆ D., DJINOVIĆ J., PETRONIJEVIĆ R.,
MILIJASEVIĆ M., JANKOVIĆ S., RADICEVIĆ T. (2009): Fatty acid
composition, cholesterol and total fat content in rainbow trout (Oncorhynchus
mykiss) as influenced by fatty acids in diet. Meat Technology 50, 179-188.
TOCHER D.R., FONSECA-MADRIGAL J., DICK J.R., NG W-K., BELL J.G.,
CAMPBELL P.J. (2004): Effects of water temperature and diets containing palm
oil on fatty acid desaturation and oxidation in hepatocytes and intestinal
enterocytes of rainbow trout (Oncorhynchus mykiss). Comparative Biochemistry
and Physiology. B: Biochemistry and Molecular Biology, 137, 49–63.
TORSTENSEN, B.E., LIE, O., FROYLAND, L. (2000): Lipid metabolism and
tissue composition in Atlantic salmon (Salmo salar L.)—effects of capelin oil,
palm oil and oleic acid–enriched sunflower oil as dietary lipid sources. Lipids, 35,
653–664.

Fatty acid profile in ...
TRBOVIĆ D., VRANIĆ D., DJINOVIĆ J., BOROVIĆ B., SPIRIĆ D., BABIĆ J.,
SPIRIĆ A. (2009): Fatty acid profile and cholesterol content in muscle tissue of
one year old common carp (Cyprinus carpio) during growth. Meat Technology 50,
276-286.
VALENTE L.M.P., BANDARRA N. M., FIGUEIREDO-SILVA A.C., REMA P.,
VAZ-PIRES P., MARTINS S., PRATES J.A.M., NUNES M.L. (2007): Conjugated
linoleic acid in diets for large-size rainbow trout (Oncorhynchus mykiss): effects on
growth, chemical composition and sensory attributes. British Journal of Nutrition,
97, 289–297.
VRANIĆ D., ĐINOVIĆ-STOJANOVIĆ J., SPIRIĆ A. (2011): Rainbow trout
(Oncorhynchus Mykiss) from aquaculture – meat quality and importance in the
diet. Meat Technology 52, 122-133.
WEAVER K.L., IVESTER P., CHILTON J.A., WILSON M.D., PANDEY P.,
CHILTON F.H. (2008): The content of favorable and unfavorable polyunsaturated
fatty acids found in commonly eaten fish. Journal of the American Dietetic
Association, 108, 1178–1185.
WILLIAMS N. (1998): Over fishing disrupts entire ecosystems. Science, 279, 809-
815.
Received 27 June 2012; accepted for publication 15 September 2012
573

Biotechnology in Animal Husbandry 28 (3), p 575-584 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 638.1
DOI: 10.2298/BAH1203575A
STUDIES ON WINGS SYMMETRY AND HONEY BEE
RACES DISCRIMINATION BY USING STANDARD AND
GEOMETRIC MORPHOMETRICS
H. F. Abou-Shaara, A. A. Al-Ghamdi
Baqshan`s Chair for Bee Research, King Saud University, P.O.Box. 2460, Riyadh 11451, Saudi
Arabia
Corresponding author: entomology_20802000@yahoo.com
Original scientific paper
Abstract: Morphometric is an essential tool for honey bee races
discrimination and characterization. Such vital tool has been applied widely in
honey bee researches. Unfortunately there is no available literature for confirming
honey bee wings symmetry. Therefore, standard and geometric morphometric
analyses were employed for investigating wings symmetry as well as for
discriminating between Carniolan and Yemeni honey bees. Moreover, three angles
of hind wings (H1, H2 and H3) were evaluated in the discrimination between the
two races. Results of morphometric analyses strongly confirmed the symmetry of
honey bee wings. Standard and geometric morphometric analyses successfully
discriminate between the two races. Hind wing length and angle H3 could be
incorporated in honey bee races discrimination.
Key words: honey bee, morphometry, symmetry, wings
Introduction
The common method for the characterization and classification of honey
bee subspecies is based mainly on measuring honey bee wing characters, which
were considered as strong tool (Rattanawannee et al., 2010). Various honey bee
colonies, races and species were discriminated by employing morphometric
analysis (Moradi and Kandemir, 2004; Raina and Kimbu, 2005; Farhoud and
Kence, 2005; Shaibi et al., 2009; Rattanawannee et al., 2010; Nedić et al., 2011).
Standard morphometric was used in honey bee studies by measuring different wing
angles, indices and distances (Ruttner, 1988) while geometric morphometric was
used in honey bee studies by measuring the coordinates of fore wing points to
calculate the centroid size (s, Tofilski, 2008). Not many studies were done on
geometric morphometric of honey bees.
Concerning hind wings, only hind wing distances and number of hooks
were used widely during morphometric studies ( Moradi and Kandemir, 2004 and

576
H. F. Abou-Shaara et al.
Abou-Shaara et al., 2012) but no hind wing angles were incorporated in the
morphometric analysis previously. Wing characters were found to be affected by
different factors e.g. temperature (Tan et al., 2005), season (Mattu and Verma,
1984) and bee age (Herbert et al.,1988).
The right fore and hind wings were used by some authors during their
morphometric analysis (Nazzi, 1992; Andere et al., 2008; Uzunov et al., 2009;
Mladenović et al., 2011 and Abou-Shaara et al., 2012) while left wings were used
by others (Bouga and Hatjina, 2005 and Tofilski, 2008). Fluctuating asymmetry
and directional asymmetry were studied by some authors (Smith et al., 1997 and
Schneider et al., 2003). Smith et al.(1997) found that honey bee drones were with
higher directional asymmetry than females while Schneider et al. (2003) found
higher fluctuation asymmetry of shape for European and hybrid workers than for
African workers. The symmetry of left and right wings is the normal hypothesis as
far there is no any malformations or abnormalities within wings. However, there
are no available literatures that can confirm honey bee wings symmetry. Thus,
standard and geometric morphometrics were employed in combination with
various computer softwares for investigating wings symmetry and for
discriminating between Carniolan and Yemeni honey bees. Moreover, three hind
wing angles were measured and evaluated for the discrimination between the two
races.
Materials and Methods
The study was done at the Bee Research Unit, King Saud University.
Thirty right and left fore wings and hind wings for two honey bee races, Yemeni
honey bees (Apis mellifera jementica) and Carniolan honey bees (Apis mellifera
carnica), were used in this study (total of 120 fore wings and 120 hind wings for
the two races). Samples of honey bee workers were collected from brood combs
and subsequently killed at -20°C and dissected by using forceps to separate fore
wings and hind wings. Separated wings were scanned by using HP scanner at 1200
dpi to obtain wing images then the following investigations were performed.
1. Symmetry of fore wings
Cubital index and discodial shift for left and right wings were measured by
Beemorph program (http://www.hockerley.plus.com/). Inner wing length, inner
wing width, distance C and distance D were measured by ScanPhoto method
according to Abou-Shaara et al. (2011) by using Photoshop program. Fore wing
angles (A1, A4, D7, J16, k19 and O26) were measured by ImageTool 3.0 program
(http://compdent.uthscsa.edu/dig/itdesc.html). Analysis of Variance (ANOVA) was
done for measured characters and means were compared by using Least Significant
Difference (P

Studies on wings symmetry and ...
Coordinates of 18 points (Figure 1) were determined by using ImageJ 1.46
program then MorphoJ program (Klingenberg, 2011) was used for geometric
morphometric analysis. Point sets of right and left fore wings were compared by
measuring centroid size, is the square root of squared distance sums of points set
from their centroid, and matrix correlation.
2. Symmetry of hind wings
Figure 1. Eighteen fore wing points
Hind wing length and width were measured by using ScanPhoto method
for right and left wings. Three hind wing angles; H1, H2 and H3 (Figure 2) were
measured by using ImageTool 3.0 program. These angles were used for confirming
the symmetry of hind wings and for discriminating between the two races.
ANOVA was performed for measured characters and means were compared by
using L.S.D.0.05.
Figure 2. Three suggested angles for hind wings
3. The discrimination between Carniolan and Yemeni honey bees
Standard morphometric was used for the discrimination between the two
races by using four lengths and six angles of right fore wings as well as by using
right hind wing characters. Forward stepwise with tolerance = 0.01, F-to-Enter:
1.000 and F-to-Remove: 0.900 was used to identify canonical discriminant
functions by using SYSTAT 13 program. Also, ANOVA was performed and
means of fore and hind wing were compared by using L.S.D.0.05 to identify
significant differences between the two races. Moreover, geometric morphometric
577

578
H. F. Abou-Shaara et al.
of fore wing points was used for the discrimination between the two races based on
their transformation grids and their centroid size.
Results and Discussion
1.Symmetry of fore wings
Mean of right wings for Yemeni honey bee was 2.54 ± 0.12 for cubital
index and -2.25 ± 0.51 for discodial shift while for left wings mean was 2.45 ± 0.11
for cubital index and -2.14 ± 0.55 for discodial shift. No significant differences
were found between cubital index and discodial shift means (L.S.D.0.05 values were
0.3259 and 1.5118 for cubital index and discodial shift, respectively). For
Carniolan honey bees, means of cubital index and discodial shift were 2.32 ± 0.05
and 2.71 ±0.38, for right fore wings, respectively and means for left wings were
2.34 ± 0.07 and 2.60 ± 0.39, respectively. No significant differences were found
between cubital index and discodial shift (L.S.D.0.05 values were 0.19 and 1.11 for
cubital index and discodial shift, respectively). Also, no significant differences
were found between studied sides in all measured lengths and angles (Table 1).
Table 1. Means ± SE of lengths and angles for Carniolan and Yemeni honey bee wings
Carniolan honey bees:
Side IWL IWW DC DD A1 A4 D7 J16 K19 O26
Right 4.37 ± 1.95 ± 0.85 ± 1.87 ± 22.30 31.39 94.61 ± 104.20 79.99 40.57
0.02 0.01 0.01 0.01 ± 0.46 ± 0.59 0.63 ± 1.25 ± 0.96 ± 1.07
Left 4.37 ± 1.94 ± 0.85 ± 1.91 ± 24.06 31.08 94.96 ± 103.39 80.59 42.23
0.02 0.01 0.01 0.01 ± 0.76 ± 0.39 0.66 ± 1.08 ± 0.88 ± 1.18
L.S.D. 0.05 0.05* 0.03 0.03 0.03 1.78 1.43 1.83 3.30 2.61 3.19
Yemeni honey bees:
Side IWL IWW DC DD A1 A4 D7 J16 K19 O26
Right 3.96 ± 1.80 ± 0.78 ± 1.73± 22.75 33.23 101.78 95.14 79.05 44.49
0.01 0.003 0.001 0.004 ± 0.80 ± 0.65 ± 0.84 ± 1.10 ±0.85 ± 1.40
Left 3.96 ± 1.81 ± 0.78 ± 1.73 ± 21.23 35.05 101.26 96.17 78.44 45.69
0.01 0.005 0.002 0.004 ± 0.69 ± 0.64 ± 0.82 ± 0.92 ± 1.16 ± 1.59
L.S.D. 0.05 0.06* 0.03 0.02 0.03 2.13 1.83 2.35 2.89 2.88 4.24
* No significant differences were found between left and right means for all characters.
IWL: Inner wing length, IWW: Inner wing width, DC: Distance C and DD: Distance D.
Centroid size for Carniolan honey bees was 0.0229 for right wings and
0.0233 for left wings and matrix correlation between matrix 1 for right wings and
Matrix 2 for left wings was 0.77. On the other side, centroid size for Yemeni honey
bees was 0.0233 for right wings and 0.0258 for left wings and matrix correlation
between matrix 1 and matrix 2 was 0.83.

2. Symmetry of hind wings
Studies on wings symmetry and ...
No significant differences were found for measured right and left hind
wing characters for Yemeni and Carniolan honey bees (Table 2).
Table 2. Means ± SE of lengths and angles for Yemeni honey bee wings
Side
Carniolan honey bees (Mean ± SE) Yemeni honey bees (Mean ± SE)
HWL HWW H1 H2 H3 HWL HWW H1 H2 H3
Right 6.17± 1.88 ± 67.41± 47.45± 27.13± 5.63± 1.82 ± 64.61± 45.41± 27.22±
0.02 0.02 1.16 0.55 0.18 0.03 0.02 1.36 0.92 0.38
Left 6.15± 1.89 ± 68.61± 47.50± 26.69± 5.61± 1.81 ± 66.50± 45.77± 26.62±
0.02 0.03 1.36 0.50 0.30 0.04 0.04 0.65 0.70 0.25
L.S.D. 0.05 0.07* 0.08 3.59 1.50 0.71 0.11 0.09 3.02 2.33 0.92
* No significant differences were found between left and right means for all characters.
HWL: Hind wing length, HWW: Hind wing width, H1: Angle H1, H2: Angle H2 and H3: Angle H3.
In general, standard morphometric analysis strongly confirmed the
symmetry of honey bee wings where no significant differences were found
between all measured characters for fore and hind wings. The most identical
characters were inner wing length, inner wing width, distance C and distance D.
Based on geometric morphometric, high degree of similarity was found between
centroid size of left and right wings for Carniolan and Yemeni honey bees.
Relatively high correlation values were found between point sets of right and left
wings for the two races. Thus the symmetry of honey bee wings was confirmed by
standard and geometric morphometric analyses. Some factors were reported to
impact on the normal wing venation pattern including pupal rearing temperature
(Tan et al., 2005), season (Mattu and Verma, 1984) and deformed wing virus
(DWV). Apart from these factors, right and left wings are identical.
3. The discrimination between the two races
Standard morphometric analysis by using fore wing characters showed that
inner wing length and width were the most successful characters in discriminating
between the two races without overlapping (Figure 3). Significant differences were
found between the two races in all measured fore wing characters except angle A1
and angle K19 (Table 3). Concerning the discrimination between the two races by
using hind wing characters, only hind wing length and angle H3 were used
successfully for the discrimination between the two races without overlapping
(Figure 4). Significant difference was found between means of hind wing length
while no significant differences were found between means of all other hind wing
characters (Table 3).
579

580
Inner wing width
2.1
2
1.9
1.8
1.7
H. F. Abou-Shaara et al.
3.7 3.8 3.9 4 4.1 4.2 4.3 4.4 4.5 4.6
Inner wing length
Races
Carniolan
Yemeni
Figure 3. The discrimination between Yemeni and Carniolan honey bees based on inner wing
length and width
H3
25.73
25.64
25.54
24.87
24.26
23.95
23.92
22.15
22.13
26
31.16
29.5
29.38
29.15
29.06
28.98
28.97
28.96
28.79
28.36
28.35
28.23
28.19
28.14
28.11
27.97
27.93
27.89
27.84
27.8
27.74
27.73
27.69
27.6
27.54
27.53
27.47
27.32
27.28
27.27
27.24
27.22
27.13
27.06
27.02
26.98
26.97
26.81
26.73
26.62
26.57
26.46
26.39
26.36
26.04
5.2
5.3
5.4
5.5
Hind wing length
5.6
5.7
5.8
5.9
6
6.1
6.2
6.3
6.4
Races
Carniolan
Yemeni
Figure 4. The discrimination between Yemeni and Carniolan honey bees based on angle H3 and
hind wing length
Table 3. Lengths and angles means ± SE for Carniolan and Yemeni honey bee wings
Race Fore wing characters (Mean ± SE)
IWL IWD DC DD Angle Angle Angle Angle Angle Angle
A1 A4 D7 J16 K19 O 26
Carniolan 4.37± 1.95 ± 0.85± 1.87± 22.30 31.39 94.61 104.20 79.99 40.57
0.02 0.01 0.01 0.01 ± 0.46 ± 0.59 ± 0.63 ± 1.25 ± 0.96 ± 1.07
Yemeni 3.96± 1.80 ± 0.78± 1.73± 22.75 33.23 101.78 95.14 79.05 44.49
0.01 0.003 0.001 0.004 ± 0.80 ± 0.65 ± 0.84 ± 1.10 ±0.85 ± 1.40
L.S.D. 0.05 0.05 0.029 0.02 0.03 1.85* 1.76 2.09 3.33 2.58* 3.53
Race Hind wing characters (Mean ± SE)
* No significant differences were found
HWL HWW H1 H2 H3 between Yemeni and Carniolan character.
Carniolan 6.17± 1.88 ± 67.41 47.45 27.13
0.02 0.02 ± 1.16 ± 0.55 ± 0.18 IWL: Inner wing length, IWD: Inner wing
Yemeni
L.S.D. 0.05
5.63±
0.03
0.09
1.82 ±
0.02
0.07*
64.61
± 1.36
3.59*
45.41
± 0.92
2.16*
27.22
± 0.38
0.85*
width, DC: Distance C, DD: Distance D.
HWL: Hind wing length, HWW: Hind wing
width, H1: Angle H1, H2: Angle H2 and H3:
Angle H3.

Studies on wings symmetry and ...
Geometric morphometric analysis was used for the discrimination between
the two races by using centroid size and transformation grid shape. Right and left
fore wings of Yemeni honey bees had higher centroid size than Carniolan honey
bees. Also, differences in transformation grids were found between fore wings of
the two races (Figure 5).
Figure 5. Transformation grids of right fore wing for Carniolan honey bees (above) and Yemeni
honey bees (below)
The effectiveness of fore wing angles in separating between honey bee
groups was identified by Nedić et al. (2011) during their work on collected honey
bee samples from different locations of Serbia. The discrimination between the two
races based on standard morphometric of fore wings showed that all measured
morphological characters, including fore wing angles, were with significant
differences between the two races except angle A1 and angle K19. Inner wing
length and width were the most successful characters in discriminating between the
two races accurately. The discrimination between the two races based on hind wing
characters showed that no significant differences were found between character
means except hind wing length. Canonical discriminant analysis showed that hind
wing length and angle H3 were used correctly for discriminating between the two
races. Thus, hind wing length and angle H3 could be incorporated in honey bee
races discrimination.
Moreover, geometric morphometric was also succeeded in discriminating
between the two races. In accordance with Tofilski (2008), who classified
successfully colonies of three honey bee races; A. m. mellifera, A. m. carnica and
A. m. caucasica by using standard and geometric morphometric and correctly
geometric morphometric identified African and European honey bees by about
99% (Francoy et al., 2008) as well as Asian honey bee species (Rattanawannee et
al., 2010). In general, standard and geometric morphometric analyses were used
successfully in the discrimination between the two races.
581

582
Conclusion
H. F. Abou-Shaara et al.
Apart from the abnormal wing venations which occasionally occur within
wings due to diseases or other factors, fore and hind wings of honey bees showed
high symmetric degree based on standard and geometric morphometric and by
employing various software programs. However, mixing right and left wings
during the morphometric analysis is not recommended. The discrimination between
the studied races was done successfully by standard and geometric morphometric
analyses. Also, MorphoJ program can be used sufficiently for performing
geometric morphometric analysis of honey bee wings as an integrated program.
Finally, hind wing length and angle H3 could be incorporated in honey bee races
discrimination.
Acknowledgment
We would like to thank the anonymous reviewers for their efforts in
improving the manuscript by their helpful comments and corrections. Also, we
would like to thank the Department of Plant Protection, College of Foods and
Agricultural Sciences & Bee Research Unit at King Saud University for providing
necessary materials for the research.
Ispitivanje simetrije krila i razlikovanja rasa medonosnih
pčela korišćenjem standardne i geometrijske morfometrije
H. F. Abou-Shaara i A. A. Al-Ghamdi
Rezime
Morfometrija je neophodan instrument u razlikovanju rasa medonosnih
pčela i njihovoj karakterizaciji i nalazi široku upotrebu u istraživanjima pčela. Na
žalost, ne postoji literatura koja može da potvrdi studije simetrije krila pčela.
Dakle, standardne i geometrijske morfometrijske analize su korišćene u analizi
simetrije krila kao i za razlikovanje medonosnih pčela rase Carniolan i Yemeni.
Štaviše, tri ugla zadnjih krila (H1, H2 i H3) su korišćena u razlikovanju rasa pčela.
Rezultati morfometrijske analize potvrđuju simetriju krila pčela. Korišćenjem
standardne i geometrijske morfometrijske analize mogu se uspešno razlikovati rase
pčela. Dužina zadnjih krila i ugao H3 se uspešno mogu uključiti u analizu i
određivanje rasa medonosnih pčela.

References
Studies on wings symmetry and ...
ABOU-SHAARA H.F., DRAZ K.A., AL-AW M., EID K. (2011): Simple
method in measuring honey bee morphological characters. Proceedings of 42 nd
International Apicultural Congress– APIMONDIA in Buenos Aries (Argentina),
21 th -25 th September, p. 222.
ABOU-SHAARA H.F., DRAZ K.A., AL-AW M., EID K. (2012): Stability of
honey bee morphological characters within open populations. Uludag Bee Journal,
12,1, 31-37.
ANDERE C., GARCIA C., MARINELLI C., CEPEDA R., RODRIGUEZ E.M.,
PALACIO A. (2008): Morphometric variables of honeybees Apis mellifera used in
ecotypes characterization in Argentina. Ecological Modelling, 214,53–58.
BOUGA M., HATJINA F. (2005): Genetic variability in Greek honey bee (A.
mellifera L.) populations using geometric morphometrics analysis. Proceedings of
the Balkan scientific conference of biology in Plovdiv (Bulgaria), 1 st -19 th May, p.
598–602.
FARHOUD H.J., KENCE M. (2005): Morphometric and MtDNA analysis in
honeybee populations (Apis mellifera L.) of north and northwest Iran. Proceedings
of the Balkan scientific conference of biology in Plovdiv (Bulgaria), 1 st -19 th May,
p. 594–597.
FRANCOY T.M., WITTMANN D., DRAUSCHKE M., MULLER S.,
STEINHAGE V., BEZERRA-LAURE M.A.F., DE JONG D., GONCALVES
L.S.(2008): Identification of Africanized honey bees through wing morphometrics:
two fast and efficient procedures. Apidologie, 39,5,488-494.
HERBERT E.W., SYLVESTER H.A., VANDENBERG J.D., SHIMANUKI H.
(1988): Influence of nutritional stress and the age of adults on the morphometrics
of honey bees (Apis mellifera L.). Apidologie, 19,3,221-230.
KLINGENBERG C.P. (2011): MorphoJ: an integrated software package for
geometric morphometrics. Molecular Ecology Resources, 11,353-357.
MLADENOVIĆ M., RENATA R., STANISAVLJEVIĆ L.Ž., RAŠIĆ S. (2011):
Morphometric traits of the yellow honeybee (Apis mellifera carnica) from
Vojvodina (Northern Serbia). Archives of Biological Sciences. 63,1,251-257.
MORADI M., KANDEMIR I. (2004): Morphometric and Allozyme Variability in
Persian Bee Population from the Alburz Mountains, Iran. Iranian International
Journal of Science, 5,2,151-166.
MATTU V.K., VERMA L.R. (1984): Morphometric studies on the Indian honey
bee, Apis cerana indica F. Effect of seasonal variations. Apidologie, 15,63-74.
NAZZI F. (1992): Morphometric analysis of honey bees from an area of racial
hybridization in northeastern Italy. Apidologie, 23,89-96.
NEDIĆ N., JEVTIĆ G., JEŽ G., ANĐELKOVIĆ B., MILOSAVLJEVIĆ S.,
KOSTIĆ M. (2011): Forewing differentiation of the honey bees from Serbia.
Biotechnology in Animal Husbandry, 27,3,1387-1394.
583

584
H. F. Abou-Shaara et al.
RAINA S.K., KIMBU D.M. (2005): Variations in races of the honeybee Apis
mellifera (Hymenoptera: Apidae) in Kenya. International Journal of Tropical Insect
Science, 25,4,281–291.
RATTANAWANNEE A., CHANCHAO C., WONGSIRI S. (2010): Gender and
Species Identification of Four Native Honey Bees (Apidae: Apis) in Thailand
Based on Wing Morphometic Analysis. Annals of the Entomological Society of
America. 103,6,965-970.
RUTTNER F. (1988): Biogeography and taxonomy of honeybees, Springer-
Verlag, Berlin.
SAS INSTITUTE (2004): The SAS System Version 9.1.3. SAS Institute,
Cary,NC.
SCHNEIDER S.S., LEAMY L.J., LEWIS L.A., DEGRANDI-HOFFMAN G.
(2003): The influence of hybridization between African and European honeybees,
Apis mellifera, on asymmetries in wing size and shape. Evolution, 57,10,2350–
2364.
SMITH D.R., CRESPI B.J., BOOKSTEIN F.L. (1997): Fluctuating asymmetry
in the honey bee, Apis mellifera: effects of ploidy and hybridization. Journal of
Evolutionary Biology, 10,551-574.
SHAIBI T., FUCHS S., MORITZ R.F.A. (2009): Morphological study of
Honeybees (Apis mellifera) from Libya. Apidologie, 40,97–105.
TAN K., BOCK F., FUCHS S., STREIT S., BROCKMANN A., TAUTZ J.
(2005): Effects of brood temperature on honey bee Apis mellifera wing
morphology. Acta Zoologica Sinica, 51,4,768-771.
TOFILSKI A. (2008): Using geometric morphometrics and standard morphometry
to discriminate three honeybee subspecies. Apidologie, 39,558–563.
UZUNOV A., KIPRIJANOVSKA H., ANDONOV S., NAUMOVSKI M.,
GREGORC A. (2009): Morphological diversity and racial determination of the
honey bee (Apis mellifera L.) population in the Republic of Macedonia. Journal of
Apiculture Research, 48,3,196 – 203.
Received 31 June 2012; accepted for publication 15 September 2012

Biotechnology in Animal Husbandry 28 (3), p 585-593 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 633.17
DOI: 10.2298/BAH1203585J
CORRELATION COEFFICIENTS OF MORPHOLOGICAL
- PRODUCTIVE TRAITS OF SPECIES OF SORGHUM
GENUS
S. Janković 2 , S. Rakić 1 , J. Ikanović 1 , J. Kuzevski 2 , Lj. Živanović 1 Ž.
Lakić 3
1
University of Belgrade, Faculty of Agriculture, Nemanjina 6, 11080 Zemun-Belgrade, Serbia.
2
Institute for Science Application in Agriculture, Bulevar despota Stefana 68b, 11000, Belgrade,
Serbia.
3
Institute of Agriculture, 78000 Banja Luka, Knjaza Miloša 17, BiH
Corresponding author: Sveto Rakić, e-mail: sveto@agrif.bg.ac.rs
Original scientific paper
Abstract: Objective of the research were phenotypic and genetic correlation
coefficients of three species of Sorghum genus – forage sorghum S. bicolor
Moench. (genotype NS-Džin), Sudan grass S. sudanense L. (genotype Zora) and
interspecies hybrid S. bicolor x S. sudanense (genotype Siloking). Studies were
carried out on samples of plant material from the first cut. The following
morphological-productive traits were studied: plant height, number of leaves per
plant, mass of leaves on the stem, average stem mass and yield of green biomass.
In the analysis of genetic and phenotypic coefficients, differences depending on the
impact on studied morphological-productive traits are observed. The highest value
of the stem height was recorded in Sudan grass (2.281 m), as well as number of
leaves per plants (7.917). The greatest mass of leaves per plant was established in
forage sorghum (49.05 g), and the highest average stem mass was recorded in
interspecies hybrid plants (80.798 g). Variation of morphological-productive
indicators per species was significant and very significant. Coefficients of simple
correlations indicate the presence of very strong to almost complete, statistically
very significant positive correlations, so these effects were expected. Plant height
and number of leaves were not directly but indirectly statistically significant to
yield of green biomass and varied from insignificant and very weak to sporadically
medium strong and statistically significant.
+
Key words: forage sorghum, Sudan grass, interspecies hybrid, genotype,
correlations.
Introduction
Plant species of Sorghum genus recently have become very interesting,
especially as forage plants, since in favorable weather conditions they regenerate

586
S. Janković et al.
well and give, depending on the moisture regime, more cuts in the year of
utilization (Glamočlija et al., 2010). Main goals of improvement and breeding of
sorghum for livestock food are to create cultivars/varieties of high production and
good quality, of long life and tolerant to limiting conditions of the environment.
Yield of green biomass and dry matter are the most important traits, which have the
utmost importance for the process of improvement/breeding, and the ultimate goal
is improvement or rarely maintaining of yields at the same level with simultaneous
improvement of other important traits. It can be improved by selection (Antoche et
al., 2007). Considering that modern programs of improvement of forage sorghum
and Sudan grass are directed towards creating of F1 hybrid with expressed potential
for high yields of green biomass of good quality and stability, suitable for use as
fresh biomass, hay or silage (Pataki et al., 2006), exceptional importance in
realization of this goal is study of the combination ability of potential parent
components (Mihajlović et al., 2007). Biomass of forage sorghum, Sudan grass and
their hybrid is used fresh or for preparation of silage, and rarely for preparation of
hay or grazing (Camakci, 1999; Ikanović et al., 2010). Depending on the way of
use of biomass (hay, silage or fresh), adequate agro-technical measures will be
applied. Of all agro-technical measures, plant nutrition is the one with the highest
impact on the quality of biomass (Booker et al., 2007). Proper application of
nitrogen mineral fertilizers shall enable better and more economical use of
environment conditions (natural factors – edaphic, climatic) and genetic yield
potential of these plants for higher production of livestock food per surface unit
(Booker 2007). Yield is reflection of the plant’s potential to accumulate dry matter,
as well as its adaptability to various agro-ecological conditions. Main criteria used
in determination of nutritional value are increase of the share of digestible matter
and reduced lignin content (Casler, 2001, Ermisani et al., 2007, Ikanović et al.,
2011). In plant improvement, it is very important to know the relation between
morphological traits, i.e. their mutual correlation (correlation coefficients) in order
to determine the improvement criteria and potential selection response of
genotypes for certain major traits (Ikanović, 2010). Direct selection for yield of dry
matter, lately considered as the most important trait from the aspect of agronomy,
in species which have been domesticated for long time, do not always give
satisfactory results, even though all modern methods are applied. So it is very
important to better understand the morphological and physiological basis of the
yield, which would make the improvement process more efficient (Sokolović,
2006, Radović, 2009). In improvement of plants, it is very important to know
relations between traits, i.e. their correlations (correlation coefficients) in order to
determine the improvement criteria and possible response of genotypes in regard to
certain major traits. The question is whether it is possible to improve plants for
individual poly-gene controlled traits. In majority of cases, undesirable changes in
values occur as well as other agronomically important traits due to mutual
correlation (Šurlan–Momirović et al., 2005). Previous studies focusing on the

Correlation coefficients of ...
chemical composition of alfalfa biomass (Milić et al., 2011) maize silage (Gocevski
and Cilev 2011) and bird’s foot trefoil (Petrović et al., 2011) indicate the
importance of this issue on nutritional properties of these feeds.
Materials and Methods
Two year research (2009-2010) was carried out on experimental field
Radmilovac. Field micro-trials were set up according to random block system in 10
repetitions with the size of main parcels of 10 m 2 (5 m x 2 m). Object of the
research were three genotypes selected in the Institute of field and vegetable crops,
Novi Sad. They are forage sorghum cultivar Džin, selected in 1983, Sudan grass
cultivar Zora selected in 1983 and interspecies hybrid Siloking, selected in 2007.
Standard agro-technics used for sorghum cropping was applied. Cutting of plants
was done in the second week of July, and samples were taken of fresh cut biomass
for analysis of morphological traits. Yield of fresh biomass was determined by
measuring of the cut above ground mass from every basic parcel and calculating
per ha. In the first year, during vegetation period, precipitation amount was by
approx. 9.5% higher compared to ten year average. April and May had less
precipitation, whereas the summer months were more humid. Precipitation amount
in the second year was higher compared to multiannual average by 27% and in
relation to the first year by approx. 20%. Distribution of precipitation during
vegetation period was equal, and maximum precipitation quantities were in June,
180 litres of rain per square meter. Distribution of heat in the first year and
according to months showed that mean monthly temperatures during summer
months were lower than multiannual average for this area. In the second year,
spring and autumn had lower air temperatures, whereas the summer was at the
level of previous year (Table 1).
Table 1. Precipitation (mm) and mean daily temperatures ( 0 C) for vegetation period, (Belgrade-
Radmilovac)
Year Parameter
IV V
Month
VI VII VIII IX
Average
Sum
2009
Temperature
Rainfall
16
6
20
34
21
153
24
79
24
45
20
45
21
362
2010
Temperature
Rainfall
14
41
18
85
21
180
24
41
24
54
18
51
20
452
Ten years Temperature 15 26 23 25 25 18 21
Average
Sum Rainfall 15 58 102 53 54 49 331
Analysis of obtained experimental data was done using analytical statistics
using statistical package STATISTICA 8 for Windows (StatSoft). By analyzing the
587

588
S. Janković et al.
covariance of investigated sorghum genotypes coefficients of phenotypic
correlation were calculated. Indicators of the mutual correlation between studied
traits were obtained from the relation of common variation and product of
individual variation. In the analysis of covariance of studied traits of investigated
sorghum genotypes, coefficients of genetic and phenotypic correlations were
calculated (Ivanović, 1984; Maletić, 2005).
Results and Discussion
Correlation relations of 6 morphological and productive traits of genotypes
of studied Sorghum species were studied on analyzed samples. Statistically
significant and very significant correlation expressed in phenotypic correlation
coefficients was established between certain traits (Table 2).
Table 2. Genetic and phenotypic correlation coefficients for morphological and productive
traits of sorghum, sudan grass and interspecies hybrid
2009
Traits
Plant height
Plant
height
Number of leaves 0.18
Number
of leaves
Leaf mass -0.8 0.44
Leaf
mass
Stem mass -0.59 0.69 0.95**
Stem
mass
Share of
leaves
Yield of
green
biomass
0.21 -0.80 -0.59 0.49 -0.33
Share of leaves 0.47 -0.78 0.90* -0.98**
Yield of green
biomass
2010
Plant height
Number of leaves 0.21
Leaf mass -0.15 0.43
0.46 0.71 -0.81 0.93**
0.96** 0.94** 0.83
-0.33 0.87* 0.83 0.95** -0.98**
Stem mass 0.60 0.69 0.95**
-0.99** 0.96**
-0.99**
0.22 -0.74 -0.49 0.59 -0.24
Share of leaves 0.47 -0.87 0.98* -0.98**
0.38 0.82 -0.84 0.93**
0.99** 0.94** 0.93
Yield of green
biomass
-0.4 3 0.97* 0.93 0.95** -0.99**
Phenotypic correlation coefficients
-0.99** 0.99**
-0.98**
Genetic correlation coefficients
Genetic correlation
coefficients

Correlation coefficients of ...
According to the correlation coefficients given in the Table 2, the values of
genetic correlation coefficients have been determined as slightly higher than the
values of phenotypic correlation coefficients in both tested years. The analysis of
morphological traits has shown a highly significant correlation between leaf mass
and stem mass, as well as between stem mass and yield of green biomass. In other
words, by increasing stem mass, leaf mass and yield of green biomass of the tested
genetic coefficients also increase. Yield of green biomass shows a highly negative
correlation (-0.99**) with share of leaves. By increasing share of leaves, yield of
green biomass decreases. Share of leaves also shows a negative correlation with
stem mass, so by increasing share of leaves, stem mass decreases. As for
phenotypic correlation coefficients, we have obtained similar, yet lower values. A
positive correlation have been determined between stem mass and leaf mass
(0.95**), as well as between yield of green biomass and stem mass. A negative
correlation has been determined between share of leaves and stem mass, and
between yield of green biomass and share of leaves. The similar results have been
obtained in the second year of testing, both with genetic and phenotypic correlation
coefficients. Statistical significance of phenotypic correlation coefficients, obtained
for certain morphological traits and their impact on yield of green biomass, is of
great importance in efforts focused on creation of new genotypes within the
species, but also for creation of interspecies hybrid. Direct selection for certain
agronomically important traits do not always end in satisfactory results, in spite of
use of all modern methods (Sokolović, 2001). Therefore, it is important to have
better understanding of the functional, morphological or physiological correlation
of traits, i.e. how and to which extent one trait influences the other and vice versa.
This is confirmed by results obtained by (Šurlan–Momirović et.al. 2005). These
authors concluded that in improvement of certain traits, controlled by majority of
genes, often undesirable changes occur in some other traits, which happens due to
mutual correlation between traits caused by association between genes.
Coefficients of simple correlations indicate the presence of very strong to almost
complete, statistically very significant positive correlations; therefore these effects
could be expected. In previous researches of mutual correlation and direct and
indirect effects of different morphological and technological traits of plant species,
deviations are very often in results obtained by simple correlation and path analysis
(Zečević, 1996; Šurlan-Momirović, 2005). This, in case of yield of green biomass
as economically most important trait, means that its expression is caused by very
complex system of various physiological and morphological indicators. Taylor
(2004) points out that analysis and study of certain morphological traits from
various aspects is necessary for defining of reliable strategy in improvement of
plant species and achieving expected results in increase of yield potential of new
genotypes. The effects of studied morphological traits on yield of green biomass of
these genotypes and their very complex mechanism of action on forming of total
589

590
S. Janković et al.
yield can be used as significant support in future work aimed at improvement of
sorghum.
Table 3. Statistical significance of differences in productive traits of sorghum cultivars
Species, Share of leaves, % Green biomass yield, t ha -1
Cultivar x ± Sx
NS Džin
Zora
Siloking
0.05
LSD 0.01
63.2 c ± 0.036
56.2 a ± 0.039
48.9 b ± 0.029
0.93
1.26
59.31 b ± 0.734
53.83 a ± 0.896
58.46 c ± 1.399
0.986
1.340
a, b, c –Values without same letter in superscript are significantly different (p

Correlation coefficients of ...
yield of green biomass of sorghum varieties. Number of leaves, as well as their
share in total biomass expressed negative effect on yield. Genotypes differed
significantly in their morphological and production traits. In the analysis of simple
correlations it was observed that correlation coefficients between plant height and
yield of green biomass, and plant height and number of leaves were very unstable
and varied from insignificant and very weak to sporadically medium strong to
statistically significant.
Acknowledgment
This work was financed by the Ministry of Education, Science and
Technical Development of, Republic of Serbia, project TR 31078.
Koeficijenti korelacije morfološko-produktivnih osobina
vrsta roda Sorghum
S. Janković, S. Rakić, , J. Ikanović, J. Kuzevski, Lj. Živanović, Ž. Lakić
Rezime
Predmet istraživanja ove studije su fenotipski i genetički koeficijenti
korelacije tri vrste roda Sorghum, i to krmni sirak S. bicolor Moench. (genotip NS-
Džin), sudanska trava S. sudanense L. (genotip Zora) i interspecies hibrid S.
bicolor x S. sudanense (genotip Siloking). Ispitivanja su vršena na uzorcima biljnog
materijala iz prvog otkosa, Proučavane su sledeće morfološko-produktivne
osobine: visina biljke, broj listova po biljci, masa listova na stablu, prosečna masa
stabla i prinos zelene biomase. Analizom genetičkih i fenotipskih koeficijenata
uočavaju se razlike i zavisnosti u delovanju na ispitivane morfološko-produktivne
osobine. Najveću vrednost visine stabla imala je sudanska trava (2,281 m), kao i
broj listova po biljci (7,917). Najveća masa listova po biljci bila je u krmnog sirka
(49,05 g), a najveću prosečnu masu stabla imale su biljke interspecies hibrida
(80,798 g). Variranja ovih morfološko-produktivnih pokazatelja po vrstama bila su
signifikantna i vrlo signifikantna. Koeficijenti prostih korelacija ukazuju na
postojanje vrlo jakih do skoro potpunih, statistički vrlo značajnih pozitivnih veza,
ovakvi efekti su se mogli očekivati. Visina biljke i broj listova nisu bili direktno
statistički značajni na prinos zelene biomase, ali indirektno jesu i varirali su od
beznačajnih i jako slabih do sporadično srednje jakih i statistički značajnih.
591

592
References
S. Janković et al.
ANTOCHE,, I. (2007): Realizari in ameliorarea sorghi la fundulea. An UNCUDA
Fundulea, LXXV, 137-157.
BOOKER, J., BRONSON , K., TROSTLE, C., KEELING, J. And MALAPATI, A.
(2007): Nitrogen and Phosphorus Fertilizer and Residual Response in Cotton-
Sorghum and Cotton-Cotton Sequences. Agronomy Journal, 99, Pp. 607-613
CASLEE M. D. (2001). Breeding forage crops for increased nutritional value.
Advances in Agronomy, 71, P 51-107.
CAMAKCI S., GUNDUZ I(1999). Effects of Different Harvesting Times on
Yield and Quality of Sorghum Silage (Sorghum bicolor L). Turkish Journal of
Agriculture and Forestry, 23, 3, 603-611.
ERISMAN, J. W.(2007): Reduced nitrogen in ecology and the environment.
Pollution, 150, 1,: 140-149
GACOVSKI, Ž., CILEV, G. (2011) Biological and genetical characteristics of
hybrid maize kneza 683A Biotechnology in Animal Husbandry 27,4,1513-1522
GLAMOCLIJA, Dj., S. JANKOVIĆ, R. MALETIĆ, S. RAKIĆ, J. IKANOVIĆ and
Z. LAKIĆ (2011): Effect of nitrogen and mowing time on the biomass and the
chemical composition of Sudanese grass, fodder sorghum and their hybrid. The
Turkish Journal of Agriculture and Forestry 35,2,127-138
IKANOVIĆ, J., Dj. GLAMOCLIJA, R. MALETIC, S. JANKOVIC, M.
TABAKOVIC, Lj. ZIVANOVIC (2010): The genotype traits of forage sorghum,
Sudan grass and their interspecies hybrid in the conditions of intensive nutrition.
Genetika, 42,2,349-358.
IKANOVIĆ, J., (2010): Genotipska i fenotipska specifičnost sorti sirka, sudanske
trave i njihovog interspecies hibrida. Doktorska disertacija, Poljoprivredni fakultet
u Beogradu, Univerzitet u Beogradu, odbrana: 20.04.2011. godine,: 116.
IKANOVIĆ, J., Đ. GLAMOČLIJA, R. MALETIĆ,., POPOVIĆ, V.,
SOKOLOVIĆ, D.,SPASIĆ, M. RAKIĆ, S. (2011): Path analysis of the productive
traits in Sorghum species. Genetika, 43,2, 253 -262
IVANOVIĆ, M. (1984): Primena metoda koeficijenata u genetičko-selekcionim
istraživanjima. Arhiv za poljoprivredne nauke, 45,160, 471-478.
MALETIĆ, Radojka (2005): Statistika. Udžbenik, Poljoprivredni fakultet,
Beograd.
MIHAJLOVIĆ, V., I. PATAKI, A. MIKIĆ, S. KATIĆ i S. VASILJEVIĆ (2007):
Dostignuća u oplemenjivanju krmnih biljaka u Srbiji Institut za ratarstvo i
povrtarstvo, Novi Sad, Zbornik radova, Sveska 44,79-86.
MILIĆ, D., KARAGIĆ, Ð., VASILJEVIĆ, S., MIKIĆ, A., MIJIĆ, B., KATIĆ, S.
(2011) Leaf and stem chemical composition of divergent alfalfa cultivars.
Biotechnology in Animal Husbandry 27,4,1505-1511

Correlation coefficients of ...
PATAKI, I., V. MIHAILOVIĆ, S. KATIĆ, S. VASILJEVIĆ, Đ. KARAGIĆ, D.
MILIĆ and A. MIKIĆ (2006): Analysis of yield components in forage sorghum
(Sorghum biocolor L.) hybrids. The Book of Abstracts of the II International
Symposium of Ecologists of the Republic of Montenegro, Kotor, Montenegro, 20-
24 September 2006, 135.
PETROVIĆ, S., VUCKOVIĆ, A., SIMIĆ, A. (2011) Stand density effects on
birdsfoot trefoil herbage yield grown for combined usage. Biotechnology in
Animal Husbandry 27,4,1523-1530
RADOVIĆ, J., SOKOLOVIĆ, D., MARKOVIĆ, J. (2009): Alfalfa-most important
perennial forage legume in animal husbandry. Biotechnology in Animal Husbandry
25,5-6,465-475
SOKOLOVIĆ, D. (2001): Genetička varijabilnost i selekciona vrednost autohtonih
populacija engleskog ljulja (Lolium perene L.), magistarski rad, Poljoprivredni
fakultet, Univerzitet u Beogradu, p.107.
SOKOLOVIĆ, D. (2006): Genetička dobit u procesu oplemenjivanja engleskog
ljulja (Lolium perene L.). Doktorska disertacija, Poljoprivredni fakultet, Univerzitet
u Beogradu, p.114.
Stat Soft (2005): STATISTICA 7.1 for Windows Inc., Novi Sad, Serbia.
ŠURLAN-MOMIROVIĆ, G., V. RAKONJAC, S. PRODANOVIĆ i T.
ŽIVANOVIĆ (2005): Genetika i oplemenjivanje biljaka – praktikum, Beograd,
231-242.
TAYLOR, J. R. N. (2004): Grain production and consumption: Africa. In: C.
Wrigley, H. Corke and C.E. Walker, Editors, Encyclopedia of Grain Science,
Elsevier, London, 70–78.
ZEČEVIĆ, V. (1996): Genetička identifikacija različitih sorti ozime pšenice
(Triticuum aesivum ssp. vulgare). Doktorska disertacija. Poljoprivredni fakultet,
Novi Sad, 5,1-98.
Received 16 May 2012; accepted for publication 15 August 2012
593

Biotechnology in Animal Husbandry 28 (3), p 595-602 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 631.563:633.11
DOI: 10.2298/BAH1203595R
QUALITY AND CONDITION OF WHEAT GRAIN
(Triticum spp.) DURING STORAGE
S. Rakić 1* , S. Janković 2 , M. Demin 1 , D. Bucalo 1 , M. Maslovarić 2
1
University of Belgrade, Faculty of Agriculture, Nemanjina 6, 11080 Zemun-Belgrade, Serbia
2
Institute of Science Application in Agriculture, Bulevar Despota Stefana 68b, 11000 Belgrade,
Serbia
Corresponding author: Sveto Rakić, e-mail: sveto@agrif.bg.ac.rs
Original scientific paper
Abstract: The effect of storage on quality and condition of grain of three
wheat cultivars (Ljiljana, Dragana and Simonida) selected in Serbia, was studied.
Freshly harvested wheat was stored at 25°C for 12 and 20 months. Decreasing of
the volume from 812.3 to 727.7 kgm -3 and absolute mass from 38.5 to 33.0% of
wheat grain during storage was recorded. Grain germination in interaction of
storage duration and temperature remained at the level above 95%. The interaction
of studied factors exhibited no statistically significant effect on the changes in the
degree of germination of wheat grain. The changes of the acid degree value (2.0–
2.4 ml 1(NaO)/100 g) were at the level of statistical significance. pH value of fresh
samples of grain was 6.5 and after 20 months 5.5. The samples tested from the
aspect of their glassiness were homogenous, the value of the coefficient of
variation for storage duration was CV

596
S. Rakić et al.
(Stanković et al., 2011) of small grains as a source of fast fermenting energy
(Vasilachil et al., 2010). In available literature, studies can be found of the effect of
temperature, air humidity and storage duration on significant losses of nutrients of
grains (Shah et al., 2002). Grain can be stored temporarily and for a longer period
of time (one or several years). Major conditions of proper storage during entire
period are air humidity and temperature. Recently, wheat grain is stored in
conditions of very low air humidity (below critical 14.5%) or in conditions in the
absence of air or lowering of temperature to the point when all life functions of
grain cease (Žeželj, 1995). Changes of physical characteristics of wheat grain
during storage have also been studied (Kent and Evers, 1994). A study (Rehman,
2006) dealing with the investigation of the effect of storage conditions on nutritive
properties of wheat, corn and rice has come to the results indicating significant
changes of the physical-chemical parameters of the tested samples. The objective
of the present study was to assess the impact of storage duration and temperature
on some physical, biological and chemical properties of wheat grain.
Materials and Methods
The tested wheat genotypes are commercial cultivars (Table 1) selected at
the Institute of Field and Vegetable Crops in Novi Sad and recorded in the Registry
of cultivars of Republic of Serbia. The material used in the analysis was produced
on the experimental field Rimski Šančevi in year 2010. The laboratory samples
were formed of freshly harvested grain using standard methods for sampling of
grain cultures (ISO 24333 : 2009) as well as adequate ICC standards and AACC
methods. All samples were protected from insects without the use of any
chemicals. Approximately 500g of each tested cultivar with 11-13% of moisture
was placed in closed plastic dishes of uniform volume. The identified samples were
stored at a controlled temperature of 25 o C for 12 and 20 months. Three dishes of
each treatment were randomly selected at the end of defined period of time and
their contents were mixed and tested in order to assess the quality and status of
samples.
Table 1. Tested wheat genotypes
Genotype Registration year Pedigree
Ljiljana 2000 NS-3287-3/Rodna
Dragana 2002 Sremka 2/Francuska
Simonida 2003 NS 63-25/Rodna//NS-3288
For assessment of the quality and state of tested wheat grain samples,
physical properties (absolute mass, volume mass, glassiness and mealy

Quality and condition of ...
breakdown), biological properties (germination ability) and chemical properties
(acid degree value and pH) were determined using standard procedures and
methods of analysis (Kaluđerski and Filipović, 1998; AOAC, 1990). pH value was
determined in the filtrate of 2g of crushed sample (1 mm mesh size) in 20 ml of
distilled water using glass electrode pH meter (Hanna, HI83141, Instruments
USA). In order to reach the objective conclusions on the effect of the observed
factors on the studied wheat grain properties, as well as the possibility of
application of parameter tests (ANOVA and LSD-test), the homogeneity of
variances was tested using the Levene′s test. The results of these tests indicated
that variances of samples in regard to almost all studied properties were
homogenous, except variances in the volume mass of wheat grain (Table 2).
Because the homogeneity was not respected, the statistical significance for higher
level of risk (1%) was observed.
Table 2. Levene′s test of samples
Properties F p-level
Glassiness and mealy breakdown 1.551 0.2092
Acid degree value 2.601 0.0505
Volume mass 5.892 0.0009
Germination ability 1.606 0.1921
Absolute mass 13.631 0.0000
pH 4.058 0.0065
The study of the differences among genotypes and storage duration, as well as their
interaction, was carried out by a two-factorial analysis of variance (ANOVA) as
well as LSD- test for the level of risk of 5% and 1% (Hadživuković, 1977). The
magnitude of the influence of each factor, as well as their interactions, was
determined by partial eta square coefficient subsequently classified according to
Cohen’s gradation (Cohen, 1988). Eta partial coefficients (η 2 ) measure a
relationship between sum of square (SS) between factors and difference between
SS total and SS error from analysis of variance (η 2 = SS between / SS total + SS
error). The relative correlation between the properties was measured with the
Pearson's correlation coefficient, at the significance level of 5% and 1%.
Experimental data were processed using statistical package STATISTICA 10 for
Windows (StatSoft).
Results and Discussion
The investigated properties of wheat grain in the analysed samples
depending on the duration and temperature of storage are presented in the Table 3.
pH values of the analysed samples were homogenous (CV

598
S. Rakić et al.
of storage. The recorded differences in pH value between these two time intervals
show a statistical significance (p

Quality and condition of ...
volume mass of fresh samples of wheat grain was the highest (812.3 kgm -3 ), and it
has been decreasing with the extension of the storage time. The recorded
differences between the storage intervals were statistically significant (p

600
S. Rakić et al.
absolute mass of grain of the genotype Simonida showed no statistically significant
difference to other two genotypes (p>0.05). The observed factors had not only a
statistically very significant effect on the change of absolute grain mass, but also
very high effect of their action, as indicated by the values of partial eta-squared
coefficients (η 2 =0.7046 and η 2 =0.3157), except that the effect of storage duration
had stronger effect than genotype.
Figure 1 illustrates changes in the acid degree value of the studied wheat
grain genotypes depending on the storage duration. The variability of the analysed
samples which accompanies the value of the acid degree value of wheat grain was
not exhibited for storage duration (CV

Quality and condition of ...
significant effect on the acid degree value, but that their effect according to
Cohen’s gradation is exceptionally high (η 2 =0.9159 and η 2 =0.8345).
Conclusion
Storage during different periods of time at the temperature of 25 o C had
negative effect on quality and condition of wheat grain. Decreasing of the values of
volume and absolute mass are registered. In addition, gradual diminishing of the
germination ability is noticeable, which, however, remained at very high level of
over 95%. The samples tested for the glassiness were homogenous, the value of
coefficient of variation for storage duration was CV

602
S. Rakić et al.
zadržala na nivou preko 95%. Interakcija ispitivanih faktora nije ispoljila statistički
značajan uticaj na promenu stepena klijavosti semena pšenice. Promene
kiselinskog stepena (2,0–2,4 ml 1(NaO)/100 g) su na nivou statističke značajnosti.
pH vrednost svežih uzoraka semena je bila 6,5 a nakon 20 meseci 5,5. Uzorci
testirani na svojstvo staklavosti su veoma homogeni, vrednost koeficijenata
varijabilnosti za dužinu skladištenja je Cv

Biotechnology in Animal Husbandry 28 (3), p 603-611 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 633.61
DOI: 10.2298/BAH1203603A
UPGRADING OF SUGARCANE BAGASSE BY SOLID
STATE FERMENTATION WITH PLEUROTUS SAJOR-
CAJU AND PLEUROTUS FLORIDA AND THE IMPACT
ON THE CHEMICAL COMPOSITION AND IN VITRO
DIGESTIBILITY
A. Akinfemi
Nasarawa State University, Faculty of Agriculture,
PMB 135 Shabu-Lafia, Nasarawa State, Nigeria.
Corresponding author: akinfemiabayomi203@yahoo.com
Abstract: Solid fermentation using celllolytic fungi: Pleurotus sajor-caju
and Pleurotus florida for upgrading of sugarcane bagasse to value-added ruminant
feed were investigated. The fermentation of the substrate lasted for 21 days after
which the changes in the chemical and mineral composition, and the in vitro gas
production were evaluated. The results obtained showed an increase in the crude
protein (%) from 6.43 (control) to 9.82 for Pleurotus sajor treated substrate (PSB)
and 10.05 for Pleurotus florida treated substrate (PFB). The treatment effect on
crude fiber, neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid
detergent lignin (ADL) was significant. Fungal treatment decreased crude fiber (%)
from 37.49 (control) to 31.67 (PFB), NDF (%) from 65.92 (control) to 53.34
(PSB), ADF (%) from 49.94 to 34.79 (PSB), and ADL (%) from 15.13 to 9.74
(PSB). Most of the major and trace minerals were higher in the untreated bagasse
with the exception of phosphorus (0.15g/kg), magnesium (1.80g/kg), potassium
(2.70g/kg) and zinc (21.60g/kg). The degradation of the insoluble but degradable
fraction (b, ml) was higher in the control (19.00) followed by PSB (16.00). The
estimated organic matter digestibility (%), short chain fatty acid (µ, mol) and
metabolisable energy (MJ/Kg DM) increased from 38.77-50.06, 0.56-0.75 and
5.33-6.80 respectively. The gas volumes at 24h, 48h and 72h as affected by
treatment was significant (P

604
Introduction
A. Akinfemi
.
Annually, huge amounts of agricultural wastes and industrial by-products
are produced worldwide from farm practices and industrial food product. In spite
of this large amounts of wastes and industrial by-products, the problems of
inadequate all year round nutrition and the prohibitive cost of conventional
feedstuffs during the dry seasons in Nigeria remained unsolved. One of such agro
industrial by-product is bagasse. Bagasse consists mainly of cellulose and
hemicellulose. Even though bagasse contains enough cellulose to make it an
excellent source of energy for ruminants, it is a poor quality feed in its natural
state. It main problems as animal feed are low digestibility, low protein content,
poor palatability and bulkiness (Abdullah et al., 2006). The poor digestibility of
bagasse is linked to the lignin complex with the cellulose thus baring it from been
accessible to rumen micro organisms. It is therefore clear that an intervention must
be applied that will overcome this limitations before it can be harnessed as
ruminant feed. Thus, considerable effort is being made to find an economic use of
this vast bulk of agricultural waste materials most of which is currently burnt as
fuel (Garg et al., 1982). In view of this, the objectives of this study were to study
the biodegradation of these wastes by Pleurotus sajor caju and Pleurotus florida
for its use as ruminant feed and the resulting impact on the chemical composition
and in vitro digestibility.
Materials and Methods
Preparation of experimental samples
Dried samples of bagasse straw were collected from the Teaching and
Research Farm, Nasarawa State University, Shabu-Lafia, Nigeria. The materials
were milled and oven-treated at 65 0 C to constant weight for dry matter
determination.
The fungus. The sporophores of Pleurotus sajor-caju and Pleurotus florida
growing in the wild were collected from University of Ibadan botanical garden.
These were tissue cultured to obtain fungal mycelia (Jonathan and Fasidi, 2001).
The pure culture obtained was maintained on plate of potato dextrose agar (PDA).
Degradation of bagasse wastes by Pleurotus sajor-caju and Pleurotus florida
Preparation of substrate.The jam bottles used for this study were thoroughly
washed, dried for 10min. at 100 0 C. 25.00g of the dried milled substrates were
weighed separately into a jam bottle and 70ml distilled water were added. The

Upgrading of sugarcane bagasse by ...
bottle was immediately covered with aluminum foil and sterilized in the autoclave
at 121 0 C for 15 min. Each treatment was in triplicates.
Inoculation. Each bottle was inoculated at the center of the substrate with 2,
10.00mm mycelia disc and covered immediately (Adenipekun and Fasidi, 2005).
They were kept in the dark cupboard in the laboratory at 30 0 C and 100% relative
humidity (RH). At day 21 day of inoculation, the experimental bottles were
autoclaved to terminate the mycelia growth. Samples of biodegradation were oven
dried to constant weight for chemical analysis and in vitro digestibility.
In vitro gas production. Rumen fluid was obtained from three West African
Dwarf female goat through suction tube before the morning feed. The animals were
fed with 40% concentrate feed (40% corn, 10% wheat offal, 10% palm kernel cake,
20% groundnut cake, 5% soybean meal, 10% brewers grain, 1% common salt,
3.75% oyster shell and 0.25% fishmeal) and 60% Guinea grass. Incubation was
carried out according to (Menke and Steingass 1988) in 120ml calibrated syringes
in three batches at 39 0 C. To 200mg sample in the syringe was added 30ml
inoculum that contained cheese cloth strained rumen liquor and buffer (9.8g
NaHCO3 + 2.77g Na2HPO4 + 0.57g KCL + 0.47g NaCL + 0.12g MgSO4. 7H20 +
0.16g CaCI2 . 2H20 in a ratio (1:4 v/v) under continuous flushing with CO2. The
gas production was measured at 3, 6, 9, 12, 15, 18, 21 and 24h. The average
volume of gas produced from the blanks was deducted from the volume of gas
produced per sample. The volume of gas production characteristics were estimated
using the equation Y = a + b (1 – e ct ) (Ǿrskov and McDonald, 1979), where Y =
volume of gas produced at time‘t’, a = intercept (gas produced from the soluble
fraction), b = gas production from the insoluble fraction, (a+b) = final gas
produced, c = gas production rate constant for the insoluble fraction (b), t =
incubation time. The post incubation parameters such as metabolizable energy
(ME, MJ/Kg DM), organic matter digestibility (OMD %) and short chain fatty
acids (SCFA) were estimated at 24h post gas collection (Menke and Steingas,
1988).
ME = 2.20 + 0.136* Gv + 0.057* CP + 0.0029*CF;
OMD = 14.88 + 0.88Gv + 0.45CP +0.651XA;
SCFA = 0.0239*Gv – 0.0601;
Where Gv, CP, CF and XA are net gas production (ml/200mg, DM) at 24 h
incubation time crude protein, crude fibre and ash of the incubated sample
respectively.
Statistical analysis. Data obtained were subjected to analysis of variance
(ANOVA) and where significant difference occurred means were separated by
Duncan (1955) using Statistical Analysis System (SAS) package.
605

606
Results and discussion
A. Akinfemi
.
The result of the chemical composition is shown in Table 1. There was
wide variation in the chemical composition. The CP (%) increased from 6.43 in the
control to 10.05 in Pleurotus florida (PFB) treated substrate while the CF (%)
decreased from 37.49 (control) to 31.47 (PFB). The increase in CP content may be
attributed to increase in microbial biomass in the form of single cell. This view was
supported by the findings of other researchers (Fasidi and Kadiri, 1993; Akinyele,
2003). Protein content increase could also be as a result of hydrolysis of starch to
glucose and its subsequent use by the same organism as a carbon source to
synthesize fungal biomass rich in protein (Bender, 1970; Hammond and Wood,
1985). Others, (Kadiri ,1999; Akinyele, 2003; Akinfemi, 2010) also reported that
protein increase may be due to secretion of certain extracellular enzymes which are
proteineous in nature into the waste during their breakdown and its subsequent
metabolism. The CF and CF fraction (NDF, ADF and ADL) decrease on the other
hand may be as a result of the ability of white-rot fungi to decompose and
metabolize all plant cell wall constituents (cellulose, hemicellulose and lignin) by
their enzymes (Erikson et al., 1990). Many species of white-rot fungi which are
effective lignin degraders have been used to assess their ability to improve the
nutritive value of fodder for ruminant nutrition (Yamakava and Okamnto, 1992;
Howard et al., 2003). Report (Chen et al., 1996) indicated that some white-rote
fungi such as the one used in this study are able to decompose free phenolic
monomers and to break the bonds with which lignin are cross-linked to the
polysaccharides. In our present study, cellulose was depleted by the fungi used
while the hemicellulose was most depleted by Pleurotus ortreatis (POB). Cellulose
and hemicellulose reduction in fungal treated substrates is common. Previous
studies (Akinfemi et al., 2010; Akinfemi and Ogunwole, 2012) support these views.
However, differences in cellulose and hemicellulose contents of POB and PFB
could be due to strain differences.
Table 1. Changes in the chemical composition and crude fiber fractions of fungal treated
Bagasse
Parameters Control PSB PFB SEM
Dry matter 89.87 c
90.54 a
90.15 b
0.006
Crude protein 6.43 c
9.82 b
10.05 a
-
Crude fiber 37.49 a
32.94 b
31.67 b
0.22
Ether extract 2.94 c
3.70 a
3.41 b
0.005
Ash 3.87 c
8.04 a
7.16 b
0.005
Carbohydrate 49.27 a
45.49 c
47.72 b
0.22
NDF 65.92 a
53.34 b
54.80 b
0.41
ADF 49.94 a
34.79 c
46.85 b
0.03
ADL 15.13 a
9.74 c
13.23 b
0.01
Cellulose 34.81 a
25.05 c
33.62 b
0.04
Hemicellulose 15.98 b
18.55 a
a,b,c, means on the same row with different superscripts are significantly varied (P < 0.05) SEM =
Standard error of mean,PSB= Pleurotus sajor-caju trated bagasse, PFB= Pleurotus florida treated
bagasse
7.95 c
0.38

Upgrading of sugarcane bagasse by ...
Table 2 shows the mineral composition of the treated substrates. The
results obtained showed that most of the major and mineral elements with the
exception of phosphorus, magnesium and zinc were higher in the untreated sample.
It is likely the microorganism might have use some of the minerals for their
metabolic activities (Akinyele et al., 2011). This observation is consistent with the
work of Frazier and Westhoff (1978) and Bannet et al., (2002) who reported that
all living organisms require some mineral elements to maintain some metabolic
functions.
Table 2. Some major minerals (g/Kg DM) and trace (ppm) mineral compositions of fungal
treated bagasse
Parameters
Major minerals
Control PSB PFB SEM
Calcium 0.70 a
0.17 b
0.22 b
0.02
Phosphorus 0.13 c
0.28 b
0.32 a
0.003
Magnesium 1.80 c
4.50 b
4.80 a
0.03
Sodium 0.80 a
0.32 b
0.35 b
0.02
Potassium 2.70 b
2.73 b
3.03 a
Trace minerals
0.05
Iron 19.20 a
15.40 c
16.37 b
0.04
Copper 10.20 a
5.40 b
4.70 c
0.03
Zinc 21.60 b
21.57 b
22.80 a
0.04
Manganese 34.50 a
13.14 c
14.40 b
0.05
a,b,c, means on the same row with different superscripts are significantly varied (P < 0.05) SEM =
Standard error of mean,PSB= Pleurotus sajor-caju trated bagasse, PFB= Pleurotus florida treated
bagasse
In Table 3, the result of gas production characteristics and estimated
metabolisable energy (ME), organic matter digestibility (OMD) and short chain
fatty acid (SCFA) is presented. Treatment effect on gas volume at 24h, 48h and
72h is significant. Gas volume ranked from the highest to lowest at 24h, 48h and
72h were POB, PFB and control. Although gas production is a nutritionally
wasteful product (Mauricio et al., 1999) it nevertheless provides a useful basis
from which ME, SCFA and OMD may be predicted (Babayemi et al., 2006).
Table 3. Gas production characteristics, and estimated metabolisable energy (ME), organic
matter digestibility OMD and short chain fatty acid (SCFA)
Parameters Control PSB PFB SEM
b (ml)
19.00 a
16.00 ab
15.00 b
0.61
C (h -1 ) 0.023 b
0.038 a
0.012 c
0.005
OMD (%) 38.77 c
50.06 a
43.42 b
0.42
SCFA (m mol)
0.56 b
0.75 a
0.59 b
0.01
ME (MJ/Kg DM) 5.33 b
6.80 a
5.86 b
0.07
Gv24 21.00 b
29.00 b
22.00 a
0.47
Gv48 38.00 b
41.00 a
41.00 a
0.33
Gv72 45.00 b
54.50 a
51.50 a
0.59
a,b,c, means on the same row with different superscripts are significantly varied (P < 0.05) SEM =
Standard error of mean,PSB= Pleurotus sajor-caju trated bagasse, PFB= Pleurotus florida treated
bagasse
607

608
A. Akinfemi
.
Menke et al., (1979) suggested that gas volume is a good parameter from
which to predict digestibility, fermentation end-product and microbial protein
synthesis.
The high volume of gas obtained in the treated substrates may be the
results of treatment effects on the cell wall content (NDF and ADF). This findings
is in agreement with the assertion elsewhere (Sallam et al., 2007) which stated that
cell wall content (NDF and ADF) were negatively correlated with gas production at
all incubation times and estimated parameters.
The potential degradation of the insoluble but fermentable fraction (b, ml)
ranged from 15 (PFB) to 19 (control). Fast rate (h -1 ) of gas production was obtained
in POB (0.038) followed by the control (0.023). The fast rate obtained in POB
could probably be influenced by carbohydrate fractions readily available to the
microbial population (Chumpawadee et al., 2007).
The higher degradation of the insoluble fraction (b ml) in the control (19.00) than
the treated bagasse, though contrary to expectations, has been previously reported
(Teguin et al., 1999; Melaku et al., 2003). Melaku et al., (2003) suggested that this
phenomenon could be due to the rapid rate of gas production leading to substrate
exhaustion and limitation on the extent of gas production in the control.
Table 4. Values for in vitro gas production (ml/200mg Dm) at different incubation period
Feed samples 3h 6h 9h 12h 15h 18h 21h 24h
Control 2 b
6 b
9 9 b
10 b
10 c
16 b
21 c
PSB 3 b
9 a
9 11 a
11 ab
13 b
16 b
29 c
PFB 7 a
8 a
8 11 a
12 a
16 a
21 a
22 b
SEM 0.27 0.19 0.27 0.33 0.33 0.19 0.69 0.47
a,b,c, means on the same row with different superscripts are significantly varied (P < 0.05) SEM =
Standard error of mean,PSB= Pleurotus sajor-caju trated bagasse, PFB= Pleurotus florida treated
bagasse
Higher values of OMD, ME and SCFA were estimated for the treated
samples. Since treatment effects on SCFA was in favour of POB and PFB
compared with the control, that suggests a potential to make energy available to
ruminants (Babayemi et al., 2006). The estimated OMD in the treated substrates is
comparable to 41.1% estimated by Sallam et al., (2007) for bagasse, but higher
than that estimated for rice straw, linseed straw and date stone. The ME estimated
for POB is high, but not significantly different between PFB and the control,
Menke and Steingass (1988) reported a strong correlation between ME values
measured in vivo and predicted from 24h in vitro gas production and chemical
composition of feed. In addition estimated ME in the present study was found to be
lower than that reported by NRC (2001).

Conclusion
Upgrading of sugarcane bagasse by ...
The fungal treated and untreated bagasse showed variation in chemical and
mineral composition, and in vitro digestibility. Based on the result obtained in this
study, the use of fungi in treatment of bagasse is a potential method to improve the
nutritional value of the substrate. However, more research is required to identify
the best fungi with the highest degradation properties. It will also be necessary to
test this feed on live animals so as to assess their response to the feed.
Unapređenje otpadaka od prerade šećerne trske čvrstom
fermentacijom sa Pleurotus sajor-caju i Pleurotus florida i
uticaj na hemijski sastav i in vitro svarljivost
A. Akinfemi
Rezime
Ispitivana je čvrsta fermentacija pomoću celulolitičnih gljiva: Pleurotus
sajor-caju i Pleurotus florida za unapređenje otpadaka u preradi šećerne trske kao
dodata vrednost hrani koja se koristi u ishrani preživara. Fermentacija supstrata je
trajala 21 dan, nakon čega su ocenjivane promene u hemijskom i mineralnom
sastavu, kao i produkcija gasa in vitro. Dobijeni rezultati su pokazali povećanje
sadržaja sirovih proteina (%) od 6,43 (kontrola) do 9,82 za Pleurotus sajor tretirane
podloge (PSB) i 10,05 za Pleurotus florida tretirane podloge (PFB). Efekat
tretmana na sadržaj sirove celuloze, NDF, ADF i ADL je bio signifikantan.
Tretman gljivama je uticao na smanjenje sirove celuloze (%) od 37,49 (controla)
do 31,67 (PFB), NDF (%) od 65,92 (kontrola) do 53,34 (PSB), ADF (%) od 49,94
do 34,79 (PSB) i ADL (% ) od 15,13 do 9,74 (PSB). Većina esencijalnih i minerala
u tragovima je imala veće vrednosti u netretiranim otpacima šećerne trske, sa
izuzetkom fosfora (0,15g/kg), magnezijuma (1,80g/kg), kalijuma (2,70g/kg) i cinka
(21,60g/kg). Degradacija nerastvorljive ali razgradive frakcije (b, ml) je bila veća u
kontrolnoj grupi (19,00), zatim PSB (16,00). Procenjena svarljivost organske
materije (%), masne kiseline kratkog lanca (μ, mol) i metabolička energija (МЈ / kg
DМ) povećana je sa 38,77-50,06, 0,56-0,75 i 5,33-6,80 respektivno. Zapremina
gasova na 24h, 48h i 72h je bila pod signifikantnim uticajem tretmana (P

610
References
A. Akinfemi
.
ADENIPEKUN C.O., FASIDI I.O. (2005): Degradation of selected agricultural
wastes by Pleurotus tuber-regium and Lentinus-Nigerian edible mushrooms.
Advances in food science, 27, 2, 61-64.
AKINFEMI A. (2010): Nutritive value and in vitro gas production of fungal treated
maize cobs. Afr. J. Food. Agri. Nutr. and Dev., 10, 8, 2943-2955.
AKINFEMI A., ADU O.A., DOHERTY F. (2010): Conversion of sorghum stover
into animal feed with white rot fungi: Pleurotus ostreatus and Pleurotus
pulmonarias. Afr. J. Biotechnol., 9, 11, 1706-1712.
AKINFEMI A., OGUNWOLE O.A. (2012): Chemical composition and in vitro
digestibility of rice straw treated with Pleurotus ostreatus, Pleurotus pulmonarius
and Pleurotus tuber-reguim. Slovak J. Anim. Sci.,45,1,14-20.
AKINYELE B.J. (2003): In vitro nutritional studies on Vovariella volvacea (Bull.
Ex Fr.) Sing, an edible mushroom, PhD Thesis, Federal University of Technology,
Akure, Nigeria, pp, 157.
AKINYELE B.J., OLANIYI O.O., AROTUPIN D.J. (2011): Bioconversion of
selected agricultural wastes and associated enzymes by Vovariella volvacea: An
edible mushroom. Research Journal of Microbiology, 6, 63-70.
BABAYEMI O.J., HAMZAT R.A., BAMIKOLE M.A., AMIRUDU N.F.,
OLOMOLA O.O. (2006): Preliminary studies on spent tea leaf: In vitro gas
production as affected by chemical composition and secondary metabolites.
Pakistan J. Nutr., 5, 5, 497-500.
BENDER, P.F. (1970): Underutilized Resources as Animal Feedstuffs. National
Academic Press, Washington D.C., pp 100.
BENNET J.W., WUCH K.G., FAISON B.O. (2002): Use of fungi in
Biomedication. In: Manual of Environmental Microbiology, 2 nd Edition, Hurst, C.J.
Crawford, R.L., Garland J.L., Lipson D.A., Mills A.L., Stretzenboch, L.D., (Eds).
ASM Press, Washington D.C., pp. 960-971.
CHEN J., FALES S.L., VARGA G.A., ROYSE D.J. (1996). Biodegradability of
free monomeric cell-wall bound phenotic acids in maize stover by two strains of
white rot fungi. J. Sci. Food Agric., 71, 145-150.
CHUMPAWADEE S., CHANTIRATIKUL A., CHANTIRATIKUL P. (2007):
Chemical composition and Nutritional Evaluation of Energy feeds for ruminant
using in vitro gas production technique. Pakistan J. Nutr., 6, 6, 607-612.
DUNCAN D.B. (1955) Multipple range and multiple F test. Biometrics, 11, 1.
ERIKSON K-E.L., BLANCHETTE K.A., ANDER P. (1990): Microbial and
enzymatic degradation of wood and wood components. Springer, Berlin,
Heidelberg, New York.
FASIDU, I.O., KADIRI M. (1993): Use of agricultural wastes for the cultivation of
Lentinus subnudus in Nigeria. Rev. Biol. Trop., 41, 411-415.
FRAZIER C.N., WESTHOFF C.D. (1978): Food Microbiology. 3 rd Edn., McGraw
Hill Inc., India, pp. 540.

Upgrading of sugarcane bagasse by ...
GARG S.K., NEELAKANTAN S. (1982): Production of SCP and cellulose by
Aspergillus terreus from bagasse substrate. Biotechnol. Bioeng, 24, 2407-2417.
HAMMOND J.W.B., WOOD D.A. (1985). Metabolism, Microbiology. In: The
Biology and Technology of the cultivated mushrooms, 2 nd Edn. Flaggy, P.B.,
Spencer D.M., Wood D.A. (Eds). John Willy and Sons, Chichester, pp. 63-80.
HOWARD R.L., ABOTSI E., VAN RENSBOURG E.L.I.J., HOWARD S. (2003):
Legnocellulose biotechnology issues of bioconversion and enzyme production. Afr.
J. Biotechnol., 2, 602-619.
JONATHAN S.G., FASIDI I.O. (2001). Effect of carbon, nitrogen and mineral
sources on growth Psathyrella atroumbonata (Pegler), a Nigerian edition
mushroom. Food. Chem, 72, 479-483.
KADIRI, M. (1999): Physiological studies of some Nigeria mushrooms. PhD
Thesis, University of Ibadan, Ibadan, Nigeria.
MAURICIO R.M., MOULD F.L., ABDALLA A.L., OWEN E. (1999): The
potential nutritive value for ruminants of some tropical feedstuffs as indicated by in
vitro gas production and analysis. Unpublished.
MELAKU S., PETERS K.J., TEGEGNE. (2003): In vitro and in situ evaluation of
selected multipurpose trees, wheat bran and lablab purpureus as potential feed
supplements of tef (Eragrostis tef) straw. Anim. Feed Sci. Technol., 108,159-179.
MENKE K.H., RAAB L., SALEWSKI A., STEINGASS H., FRITZ D.,
SCHNEIDER W. (1979): The estimation of the digestibility and metabolizable
energy content of ruminant feeding stuff when they are incubated with rumen
liquor. J. Agri. Sci., 93, 217-222.
MENKE K.H., STEINGASS. (1988): Estimation of the energetic feed value
obtained from chemical analysis and in vitro gas production using rumen fluid.
Anim. Res. Dev., 28, 7-55.
NRC. (2001): Nutrient requirements of dairy cattle (7 th Rev Ed). National research
council, National Academy Press. Washington, D.C.
ǾRSKOV E. R., MCDONALD L. M (1979): The estimation of protein
degradability in the rumen from incubation measurement weighted according to
rate of passage. Journal of Agricultural Science (Cambridge), 92, 499-503.
SALLAM S.M.A., NASSER M.E.A., EL-WAZIRY A.M., BUENO I.C.S.,
ABDALLA A.L. (2007): Use of an in vitro gas production technique to evaluate
some ruminant feedstuffs. J. Appl. Sci. Res., 3,1, 34-41.
TEGUIN A., ORSKOV E.R., KYLE D.I. (1999): A note on ruminal insutu
degradability and in vitro gas production of some West African grass species and
multipurpose legume tree leaves. J. Anim. Feed Sci., 8,415-424.
YAMAKAVA M., OKAM NTO H.A. (1992): Effect of incubation with edible
mushroom, Pleurotus ostreatus, on voluntary intake and digestibility of rice bran
by sheep. Anim. Feed Sci. Technol., 63,133-138.
Received 30 July 2012; accepted for publication 3 August 2012
611

Biotechnology in Animal Husbandry 28 (3), p 613-622 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 631.82
DOI: 10.2298/BAH1203613K
INFLUENCE OF MINERAL FERTILIZATION ON SOME
BIOLOGICAL AND PRODUCTIVE INDICATORS OF
NATURAL MEADOW OF AGROSTIS CAPILLARIS-
FESTUCA FALLAX TYPE IN THE RHODOPE
MOUNTAINS (SOUTHERN BULGARIA)
Ya. Kozhouharov 1 , V. Lingorski 2
1 Complex Experimental Station, 4700 Smolyan, Bulgaria
2 Research Institute of Mountain Stockbreeding and Agriculture, 5600 Troyan, Bulgaria
Corresponding author: vilievl@yahoo.com
Original scientific paper
Abstract: The experiment with natural meadow of Agrostis capillaris -
Festuca fallax type in Rhodope Mountains (Smolyan region, Southern Bulgaria)
shows that according to fertilization variants there is a clear tendency towards
earlier start of active vegetation and its next phenophases. Most and almost equal
amounts of dry biomass (4.38 and 4.31 t.ha -1 ) were reported in fertilization with
N160P80K80 and N160P80, which exceeded the unfertilized control by 128.12 and
124.48%, respectively. Compared with unfertilized variant, the crude protein
content increased in all fertilizing variants, as in complete mineral fertilization with
N80-160 accumulates the crudest protein - respectively 314 g.kg -1 and 318 g.kg -1 . As
regards to other studying chemical indicators of the forage (crude fiber, crude fat,
crude ash and nitrogen-free extract substances) the mineral fertilization also had an
expressed positive effect.
Key words: meadow, Agrostis capillaris-Festuca fallax type, fertilization,
bioproductivity, the Rhodope Mountains.
Introduction
The meadows and pastures in the mountainous regions of Bulgaria have a
highly productive potential as a basic and only one source of obtaining roughage
for refute or pasture breeding of animals (mainly sheep and cattle).
The natural meadows of Agrostis capillaris-Festuca fallax type in Bulgaria
occupy an area of 50 783.3 ha, which is 22.66% of the total area of natural
meadows in the country (224 145.8 ha). The natural pastures of this type amounted
to 82 696 ha, which is 6.76% of the total area of natural pastures in the country
(1 222 896.2 ha). In Smolyan region natural meadows of this type reached 56.8%
of the total area of natural meadows in the region,and annually give about 1.5-2.0

614
Ya. Kozhouharov et al.
t.ha -1 with good quality hay (in 100 kg hay containing 46 food unit and 3.41 kg
digestible protein). The natural pastures of the same type here reached 39.2% of the
total area of natural pastures in the region, giving an annual yield of 3.6-4.4 t.ha -1
green mass with good quality feed. (Cheshmedjiev, 1976; Yakimova et al., 1977).
Due to irrational use and low levels of applied agricultural machines, the
condition of natural meadows and pastures in the region is unsatisfactory.
Furthermore, the main reasons for their low forage productivity are specific
vegetation of grass stands and available natural and environmental characteristics
that are typical of the region such as weak soil reserve with essential nutrients,
high soil acidity, irregular rainfall distribution, a high rough of country, etc.
The conducted multiplied studies in Bulgaria (Totev, 1984; Pavlov, 1996
and others) and abroad (Kasper, 1971; Sur, 1975; Sung and Kim, 1985; Grandi et
al., 1989; Giraldez et al., 1993 and others) show that apart from specific natural
characteristics the quality and quantity of biomass obtained from natural grasslands
has been also influenced to a great extent by the level of enforcement. Mineral
fertilization and its ways of use are among the most important in farming practices.
The purpose of this study was to identify the changes in productivity and
chemical composition of forage biomass from natural meadow of Agrostis
capillaris-Festuca fallax type in the Rhodope Mountains (near the town of
Smolyan, Southern Bulgaria) under the influence of mineral fertilization with
different rates and combinations of nitrogen, phosphorus and potassium fertilizers
and dynamic of grass biomass accumulation during a period of vegetation and in
different phenophases of grass growth.
Materials and Methods
The field experiment was conducted during the 1992-1994 period, on
natural meadow of Agrostis capillaris-Festuca fallax type in the Rhodope
Mountains (near the town of Smolyan, Southern Bulgaria) at 1100 m altitude. The
soil in the area of experiment was a brown forest with light mechanical structure
because the chemical composition was characterized by a middle reserve of humus
and a low total nitrogen and phosphorus. Low values were established by water
soluble forms of nitrogen, phosphorus and molybdenum and the reserve of water
soluble forms of potassium and boron was optimum. The soil reaction was acidic.
The block-method was used in four repetitions and the harvesting plot area
of 25 m 2 with the following fertilizer rates in kg per 1 ha as variants: 1. N0P0K0
(unfertilized variant) – Control; fertilization variants: 2. N80; 3. P80; 4. K80; 5.
N80P80; 6. N80K80; 7. P80K80; 8. N80P80K80; 9. N160; 10. N160P80; 11. N160K80; 12.
N160P80K80. The fertilization was accomplished annually, in early spring and
shortly before the beginning of active vegetation of grasses, with nitrogen (as
ammonium nitrate), phosphorus (as superphosphate) and potassium (as potassium

Influence of mineral fertilization ...
sulphate). The trial plots were hay-making by hand in full flowering phenophase of
grasses.
During the conduction of the field experiment information about the
following indicators was collected:
1. Time for passing of the main phenophases of grasses - through visual
observation by Rudenko-method.
2. Growth swiftness of grasses – such as complete stalk shooting, ear-formation
and full flowering phenophases – by date.
3. Dry mass (DM) yields (in t.ha -1 ) - it was established by drying constant weight
at 105 0 C of 0.5 kg green mass samples, taken immediately after cutting each trial
plot and repetition. It was reported for years and average for the experimental
period.
4. Chemical composition of the absolutely dry matter – crude protein content (by
Kieldahl-method); of crude fiber (by Kyushner and Haver-method); of crude fat –
by ether extract; of nitrogen-free extract substances (NFES) – the amount
difference among crude protein, crude fiber, crude fat and crude ash; of crude ash –
by weighing.
The daily temperatures for each of the months in the Smolyan region for
the 1977-1996 period characterize the region by mild winters, cool springs and
summers without too much heat and with relatively warm autumn months. The
temperature regime significantly influenced the growth of grasses the duration of
vegetation period as well as the accumulation intensity of green mass and dry
matter.
For the same 20-annual period the humidity was relatively high and annual
rainfalls (1180.2 l/m 2 ) were almost two times more than the average amount of the
country (620 l/m 2 ). The rainfall is unevenly distributed. There is clearly marked
winter and spring-summer rainfall maximum, as rainfalls in April, May and June
are crucial for the natural grasslands productivity. The rainfall reduction during
July, August and September has a negative impact on productivity of grasslands.
Results and Discussion
Time for passing of the main grass phenophases. From Table 1 it is
obvious, that in 1992 the growing-up phenophase in the control occurred on 05.05.,
while self-fertilization with nitrogen (regardless of its rate), with phosphorus or
potassium, in combined fertilization (var. 5-7 and var. 10-11) and in complete
mineral fertilization (var. 8 and 12) – on 02.05.-03.05. Similar differences in
control were observed in other stages. So, compared to fertilization variants the
stalk shooting in var. 1 also occurred later – on 08.06., as against 05.06.-07.06. in
self- and combined fertilization with lower rates (80 kg.ha -1 ) and on 01.06.-02.06.
in var. 8-12. That same year, the ear-emergence stage in the control also occurred
later – only on 30.06., while in other variants varied from 20.06. tо 22.06. (in
615

616
Ya. Kozhouharov et al.
complete mineral fertilization as well as self- and combined fertilization with N160).
In self- and combined fertilization with lower rates this stage occurred towards
25.06.-28.06.
Таble 1. Time for passing of the main phenophases of grasses by years and dates.
Fertilization
variants
Growing-up Stalk shooting Ear-formation Flowering
1992 1993 1994 1992 1993 1994 1992 1993 1994 1992 1993 1994
1.N0P0K 0(Control) 05.05. 07.05. 08.05. 08.06. 07.06. 10.06. 30.06. 27.06. 04.07. 22.07. 23.07. 24.07.
2. N80 03.05. 04.05. 05.05. 05.06. 04.06. 07.06. 25.06. 23.06. 28.06. 20.07. 19.07. 21.07.
3. Р80 03.05. 05.05. 06.05. 07.06. 07.06. 08.06. 28.06. 25.06. 03.07. 22.07. 22.07. 23.07.
4. К80 02.05. 05.05. 07.05. 07.06. 06.06. 08.06. 28.06. 26.06. 04.07. 23.07. 20.07. 22.07.
5. N80P80 03.05. 04.05. 05.05. 05.06. 04.06. 06.06. 22.06. 23.06. 26.06. 18.07. 17.07. 19.07.
6.N80K80 02.05. 03.05. 05.05. 06.06. 05.06. 06.06. 26.06. 24.06. 27.06. 22.07. 20.07. 19.07.
7. Р80К80 02.05. 05.05. 07.05. 07.06. 07.06. 08.06. 28.06. 25.06. 03.07. 21.07. 21.07. 22.07.
8.N80P80K80 02.05. 02.05. 03.05. 02.06. 02.06. 03.06. 22.06. 23.06. 23.06. 14.07. 15.07. 16.07.
9. N160 02.05. 03.05. 03.05. 01.06. 02.06. 01.06. 22.06. 22.06. 23.06. 18.07. 19.07. 18.07.
10. N160P80 03.05. 03.05. 04.05. 01.06. 01.06. 30.05. 21.06. 21.06. 21.06. 15.07. 13.07. 13.07.
11. N160K80 02.05. 03.05. 03.05. 02.06. 01.06. 01.06. 22.06. 22.06. 22.06. 14.07. 15.07. 15.07.
03.05. 03.05. 04.05. 01.06. 01.06. 02.06. 20.06. 21.06. 21.06. 13.07. 15.07. 14.07.
12.N 160P 80K 80
The full flowering stage occurred later in fertilization with K80, Р80, N80K80,
Р80K80 and in unfertilized control – between 21.06.-23.06., and earliest in complete
mineral fertilization (var. 12) and combined fertilization with N80-160 – during the
13.06.-18.06. period. Тhus, fertilization with N80P80K80, N160K80 and N160P80 reduced
the time from grow-up beginning to full flowering of 7 days, and fertilization with
N160P80K80 - of 8 days. From Table 1 it is clear, that generally self-fertilization with
nitrogen, phosphorus or potassium does not lead to significant differences needed
for the main stages of grasses.
During next year (1993) unfertilized control phenophases occurred later than
fertilization variants. As a result, the start of the grow-up in control occurred on
07.05., while the early onset of vegetation was found in fertilization with N80P80K80
– still on 02.05.
The combined fertilization with lower and higher rates and complete
fertilization with N160P80K80 caused vegetation on 03.05., while self- and combined
fertilization with N80P80 – on 04.06.-05.06. The next phenophase - the stalk shooting
occurred later in the control and fertilization with Р80 and P80K80 – on 07.06., and
earliest in combined and complete mineral fertilization with N160 – still on 01.06.,
and other variants occupied an intermediate position.
The earliest ear-formation stage again occurred in self-, combined and
complete mineral fertilization with N160 – on 21.06.-22.06., аnd later again in
unfertilized control – barely on 27.06. In other fertilizing variants this stage

Influence of mineral fertilization ...
occurred in the period 23.06.-26.06. The earliest full flowering stage was
established in fertilization with N160P80 – still on 13.07., followed by var. 8, 11 and
12 – on 15.07., but the latest was the flowering of control variant – 8 days later (on
23.06.).
The other studying variants were in an intermediate position in relation to
the occurrence of that stage. It is seen that in 1993 the complete mineral
fertilization (вар. 8 and 12) reduced the time from growing-up to full flowering by
3 and 4 days, combined fertilization with N80P80 (var. 5) – by 3 days, combined
fertilization with N160 (var. 10 and 11) – by 6 and 3 days, respectively, while selffertilization
(regardless of norm) occupied an intermediate position.
In the last year of study (1994) the growing-up phenophase again occurred
later in the control variant – at 08.05., followed by self-fertilization with К80 and
combined fertilization with P80K80 – of 07.05. The earliest occurrence of this stage
was reported in fertilization with N80P80K80, N160 and N160K80 – of 03.05., while the
other studied variants ranged from 04.05. (var. 10 and 12) tо 06.05. (var. 3). The
next phenophase (the stalk shooting) occurred earliest (as early as 30.05.) in
fertilization with N160P80, followed by var. 9 and 11 (fertilization with N160 and
N160K80) – to 01.06., and latest again in unfertilized control – only 10.06. In other
variants this stage occurred between the periods 02.06. (var. 12) – 08.06. (var. 3, 4
and 7). In most studying variants (eight) the next phenophase (the ear-formation)
occurred in June (more on 21.06.) and it was established in fertilization with
N160P80 and N160P80K80, and latest in control and self-fertilization with K80 – only
04.07., followed by fertilization with Р80 and P80K80 – on 03.07., while the other
variants occupied an intermediate position regarding the occurrence of this
phenophase. The same year, the full flowering occurred earliest after the complete
mineral fertilization with N160P80 - still on 13.07., and latest (after 11 days) in
control – 24.07., followed by self-fertilization with nitrogen, phosphorus or
potassium (var. 2, 3 and 4) and combined fertilization with Р80К80 – between 21.06
and 23.06. The same table shows that most of the time from the start of the
growing-up phenophase to full flowering stage was reduced in fertilization with
N160P80 – with 7 days, followed by complete mineral fertilization with N160P80K80
and N160K80 – by 6 and 4 days, respectively.
In generalization, differences in the occurrence of the main phenophases
are not only due to differences in application of mineral fertilizers (such as
combinations and rates), but the existing weather conditions during the vegetation
period of the correspondingly year. However, during the study, the fertilizing
variant observed a clear trend towards earlier initiation of active vegetation and
hence to an earlier onset of the next phenophase of grasses, although the
predominant species in grass stand (Agrostis capillaris) is characterized by a slow
growth.
Productive potential of the grass area.The results from Table 2 show that
compared with unfertilized control the grass area productivity exceeded
617

618
Ya. Kozhouharov et al.
significantly in the studying fertilization variants as early as the first year of the
experiment (1992). Thus, the obtained dry mass in self-fertilizing variant with
nitrogen, phosphorus or potassium in rates 80 kg.ha -1 (var. 2, 3 and 4) exceeded
unfertilized variant by 54.59, 35.20 and 14.79%, respectively. The combined
fertilization with the same rates (var. 5, 6 and 7) increased in greater yields
compared to the control – by 79.08, 68.37 and 55.10%, while the complete
fertilization (var. 8) was double the excess – with 104.08%. Doubling the rate of
nitrogen (160 kg.ha -1 ), despite its own, double or triple combination (var. 9-12)
resulted in an increase of dry biomass obtained from 4.04-4.41 t.ha -1 , which was
more 106.12-125.00% compared with unfertilized control.
Table 2. Dry matter yields (t.ha -1 ) by years and average for the 1992-1994 period.
Fertilization 1992 1993 1994 Average Proof
variants
for the period
t.ha -1
% t.ha -1
% t.ha -1
% t.ha -1
%
1. N0P0K0 (Control) 1.96 100.00 1.73 100.00 2.07 100.00 1.92 100.00 -
2. N80 3.03 154.59 2.82 163.00 2.96 142.99 2.94 153.12 +
3. Р80 2.65 135.20 2.46 142.20 2.61 126.09 2.57 133.85 -
4. К80 2.25 114.79 2.02 116.76 2.11 101.93 2.13 110.94 -
5. N80P80 3.51 179.08 3.64 210.40 3.76 181.64 3.64 189.58 +++
6. N80K80 3.30 168.37 3.16 182.66 3.27 157.97 3.24 168.75 ++
7. Р80К80 3.04 155.10 2.91 168.21 2.98 143.96 2.98 155.21 +
8. N80P80K80 4.00 204.08 3.98 230.06 4.08 197.10 4.02 209.37 +++
9. N160 4.04 206.12 3.90 225.43 4.08 197.10 4.00 208.33 +++
10. N160P80 4.27 217.86 4.11 237.57 4.57 220.77 4.31 224.48 +++
11. N160K80 4.09 208.67 4.03 232.95 4.18 201.93 4.10 213.54 +++
12. N160P80K80 LSD 0.05
23.7
LSD 0.01
34.5
4.41 225.00 4.22 243.93 4.50 217.39 4.38 228.12 +++
The same table shows that over the next year (1993) dry mass yields
decreased in all variants compared with the previous year. So in the control variant
there was 1.13 times decrease, in self-fertilizing with nitrogen, phosphorus or
potassium in rates 80 kg.ha -1 – from 1.07 to 1.11 times, in the combined
fertilization with the same rates (var. 6 and 7) – 1.04 times, in complete
fertilization (var. 8) – 1.00 time. Doubling the rate of nitrogen, whether selfdependent,
double or triple combination (var. 9-12) decrease was from 1.01 to 1.04
times. The excess compared to the control in the obtained dry mass varied by type
and rates of fertilizers. Thus, in self-fertilization (var. 2, 3 and 4) was less and
amounted to 63.00, 42.20 and 16.76%, in combined fertilization with N80K80 and
Р80К80 reached 82.66 and 68.21%, whereas combined fertilization with N160Р80K80

Influence of mineral fertilization ...
was the more - over a 143.93%. The other variants in combination with N160
occupied an intermediate position.
In the last year of study (1994) dry mass yields increased compared to
those that were obtained in 1993 as follows: 1.20 times under control, by 1.04 to
1.06 times in self-fertilizing with nitrogen, phosphorus or potassium at a rate 80
kg.ha -1 , 1.02 to 1.03 times in combined fertilization with the same rates, while
increasing the nitrogen alone, or combined with a comprehensive introduction
phosphorus and/or potassium reached by 1.04 to 1.11 times. Nearly identical and
maximum yields for the year were obtained by fertilization with N160P80 (4.57 t.ha -
1 ) and N160P80K80 (4.50 t.ha -1 ), as their excess in comparison with the unfertilized
control reached 120.77 и 117.39%. Relatively small excess was in self-fertilizing
with nitrogen, phosphorus or potassium in rates 80 kg.ha -1 (var. 2, 3 and 4) –
respectively 42.99, 26.09 and only 1.93%, and in combined fertilization in the
same rates (var. 5, 6 and 7) – respectively 81.64, 57.97 and 43.96%. The other
variants of fertilization occupied an intermediate position in the dry mass
productivity.
Average for the experimental period (1992-1994) nearly equal amounts of
dry biomass were obtained after fertilization with N160P80K80 and N160P80 – 4.38
and 4.30 t.ha -1 , this exceeded the unfertilized control respectively 128.12 and
124.48%. Significantly (by 109.37%) the dry matter yields were increased in
complete mineral fertilization with N80P80K80 (var. 8).
It is evident that nitrogen self-fertilizing increased the dry mass
productivity of natural meadow Agrostis capillarys - Festuca fallax type, while the
self phosphorus and potassium fertilization did not cause significant changes in the
yield of dry matter. To a certain extent the effectiveness of potassium was
increased in combination with N80 and significant in combination with N160. On the
productivity of dry matter combined phosphorous-potassium, nitrogen-potassium
and nitrogen-phosphorous fertilizers increased the yields and the differences
compared with unfertilized variant were well and very well demonstrated (++ ;
+++). The complete mineral fertilization in rates of 80 and 160 kg.ha -1 nitrogen
together with phosphorus and potassium increased productivity of grass stand in
significantly higher compared with their self or combined use. The complete
mineral fertilization with N80P80K80 was less effective in terms of dry matter yield
compared to fertilization with N160P80. This showed that increasing the rates of
nitrogen had significantly greater effect on the addition of potassium in the lower
nitrogen part in fertilizer combination. It is clear, that in combination with different
mineral fertilizers potassium inhibited the effects of nitrogen and phosphorus part.
Chemical composition of the grass biomass The changes in the chemical
content of forage biomass (Table 3) show that on average during the study
compared with unfertilized control the crude protein content in dry matter
increased in all fertilized variants. It is clear that the complete fertilization as a N80,
619

620
Ya. Kozhouharov et al.
and with N160, aroused the highest crude protein content – respectively 314 g.kg -1
and 318 g.kg -1 . The combination of nitrogen and potassium also added to significant
amounts of crude protein - 312 g.kg -1 at fertilization with N80K80 and 314 g.kg -1 at
fertilization with N160K80. Although the combination with nitrogen, phosphorus and
potassium increased the nutritional value of dry matter, they did not cause a
significant increase in the crude protein content. Regardless of the size of the
fertilizer rate (80 or 160 kg.ha -1 ), the lowest values of crude protein in dry matter
were recorded in nitrogen self-fertilization – respectively 248 and 267 g.kg -1 , as
well as phosphorus and potassium self-fertilization - 276 and 288 g.kg -1 .
The complete mineral fertilization with N80-160 lower in most fiber content in
dry matter – corresponding to 249 and 256 g.kg -1 and the unused of fertilizers (as it
in control) and self-fertilization or in combinations with them increased reaching
280 g.kg -1 (fertilization with N80P80) to 318 g.kg -1 (fertilization with N80) - 333 g.kg -
1 (control).
Таble 3. Chemical composition of the grass biomass (g.kg -1 DM) average for the 1992-1994
period.
Fertilization
variants
Crude protein Crude fiber Crude fat NFES Crude ash
1. N0P0K0 (Control) 185 333 50 369 63
2. N80 248 318 51 341 42
3. Р80 276 290 53 348 33
4. К80 288 294 53 345 20
5. N80P80 305 280 52 340 23
6. N80K80 312 261 52 340 35
7. Р80К80 306 263 53 349 29
8. N80P80K80 314 249 54 320 63
9. N160 267 284 53 328 68
10. N160P80 312 270 54 310 36
11. N160K80 314 265 54 312 55
12. N160P80K80 318 256 55 316 55
The same table shows that regardless of unused or fertilizer application,
crude fat varied in relatively small limits, increasing from self-fertilization to the
combined fertilization and from lower to higher rates of fertilizers. Thus, the least
crude fat (50 g.kg -1 ) were found in the dry matter of unfertilized control, of selffertilization
with nitrogen, phosphorus or potassium (var. 2-4) gradually increased
to 51-53 g.kg -1 , the combined fertilization (var. 5-7) – 52-53 g.kg -1 , the combined
fertilization with N160 (var. 10-11) – 54 g.kg -1 , and the complete mineral
fertilization (var. 12) were the most - 55 g.kg -1 .
Regarding the content of nitrogen-free extract substances (NFES) opposite
tendency was observed as the highest value (369 g.kg -1 ) was reported in control.
With the application of mineral fertilizers decreased values in self-fertilization (var.

Influence of mineral fertilization ...
2-4) - 341-348 g.kg -1 and the combined fertilization (var. 5-7) - 340-349 g.kg -1 to the
complete fertilization (var. 12 and 8) - 316-320 g.kg -1 .
The crude ash content varied in relatively wide ranges, the maximum value
reached during fertilization with N160, the complete fertilization (var.8) as well as
control – respectively 68, 63 and 63 g.kg -1 . At least crude ash in dry matter was in
fertilization with К80, N80P80 and Р80К80 – 20, 23 and 29 g.kg -1 , and other variants
occupied an intermediate position.
Conclusion
The experiment with a natural meadow of Agrostis capillaris-Festuca
fallax type in Rhodope Mountains (Smolyan region, Southern Bulgaria) shows that
according to fertilization variants there is a clear tendency towards earlier start of
active vegetation and its next phenophases.
Most and almost equal amounts of dry biomass (4.38 and 4.31 t.ha -1 ) were
reported in fertilization with N160P80K80 and N160P80, which exceeded the
unfertilized control by 128.12 и 124.48%, respectively.
Compared with unfertilized variant, the crude protein content increased in
all fertilizing variants, as in complete mineral fertilization with N80-160 accumulates
the crudest protein - respectively 314 g.kg -1 and 318 g.kg -1 .
As regards to other studying chemical indicators of the forage (crude fiber, crude
fat, crude ash and nitrogen-free extract substances) the mineral fertilization also
had an expressed positive effect.
Uticaj mineralnog dubrenja na neke biološke I proizvodne
indikatore prirodnih livada tipa Agrostis capillaris-festuca
fallax u rodopskim planinama (južna Bugarska)
Ya. Kozhouharov, V. Lingorski
Rezime
Ogled na prirodnim livadama tipa Agrostis capillaris - Festuca fallax u
rodopskim planinama (oblast Smoljan, južna Bugarska), prema varijantama
đubrenja, pokazuje jasnu tendenciju ranije početka aktivne vegetacije i njenih
kasnijih feno-faza.
Utvrđene su skoro identične količine biomase (4.38 and 4.31 t.ha -1 ) u
varijantama đubrenja sa N160P80K80 i N160P80, sa vrednostima iznad kontrole bez
đubrenja od 128,12 i 124,48%, respektivno.
621

622
Ya. Kozhouharov et al.
U poređenju sa neđubrenom varijantom, sadržaj sirovog proteina se
povećao u varijantama sa đubrenjem, jer se u slučaju potpunog N đubrenja sa N80-
160 akumulira sirovih proteina – respektivno 314 gkg -1 i 318 gkg -1 .
U vezi sa ostalim ispitivanim hemijskim parametrima krmiva (sirova
celuloza, pepeo i bezazotne materije), mineralno đubrenje je imalo pozitivan uticaj
i na njih.
References
CHESHMEDJIEV B. (1976): Feed characteristics of the natural pastures and
meadows in Bulgaria. Dissertation on PhD of Agricultural Sciences, Sofia.
GIRALDEZ F., ALVARES J., MARTINES F., SOLIS S. (1993): Ruminal
degradation of hay from permanent pastures. Effect of stage of maturity, Archives
de Zootechnica, 42, 156, 13-20.
GRANDI A., CAGIOTTI M., BLASSI F. (1989): Investigation of flora
productivity and nutritive value of meadow pasture in Ragnolo Macerata.
Zootechnica e Nutrizione Animale, 15, 2, 115-133.
KASPER J. (1971): Prosperok k studinzmien v chemickom zloteny docasnych
travnicu parastov pri razdielny dusteatej vyzive. Redecke Prace, VULP Banska
Bistrica, 6, 111-137.
PAVLOV D. (1996): Productivity, nutritive value, qualitative characteristics of
different groups forage plants and possibilities for their prediction. Dissertation on
PhD of Agricultural Sciences, Stara Zagora.
SUNG K., KIM C. (1985): Effect of intake on digestibility of grass hay harvested
at different cutting dates, Journal of the Korean Society of Grassland Science, 5, 2,
111-115.
SUR D. (1977): Vpliv vizivy a viuzivania na dinamiku narastania hmoty na
prirodzenom travnom parasite. Redecke Prace, VULP Banska Bistrica, 9, 49.
TOTEV Т. (1984): Investigations on the regeneration and utilization of natural
meadows and pastures in the upland mountainous and alpine regions of central part
of the Balkan range. Dissertation on PhD of Agricultural Sciences, Troyan.
YAKIMOVA YA., OUZUNOV M., PETROVSKI N., POPOV I., MITEV M.
(1977): Improvement and utilization of natural meadows and pastures. Zemizdat,
Sofia, pp. 282.
Received 18 June 2012; accepted for publication 16 August 2012

Biotechnology in Animal Husbandry 28 (3), p 623-633 , 2012 ISSN 1450-9156
Publisher: Institute for Animal Husbandry, Belgrade-Zemun UDC 636.09
DOI: 10.2298/BAH1203623D
CONTROL OF BROWN RAT (Rattus norvegicus) ON A
DAIRY FARM IN SERBIA
S. Đedovic¹, M. Vukša 1 , M. M. Petrović 2 , J. Bojkovski 3 , I. Pavlović 4 , G.
Jokić¹, B. Stojnić 5
1
Laboratory of Applied, Institute of Pesticides and Environmental Protection, Banatska 31b,
Belgrade, Serbia
2
Institute for Animal Husbandry, Autoput 16, Belgrade, Serbia
3
University of Belgrade, Faculty of Veterinary Medicine, Bulevar oslobođenja 18, Belgrade, Serbia
4
Scientific Institute of Veterinary Medicine of Serbia, Belgrade, V. Toze 14, Belgrade, Serbia
5
University of Belgrade, Faculty of Agriculture, Nemanjina 6, Belgrade, Serbia
Corresponding author: Suzana.Djedovic@pesting.org.rs
Original scientific paper
Abstract: Rattus norvegicus is a synanthropic species living almost
exclusively around facilities for keeping domestic animals. This three-year
research focused on options for reducing economic damage caused by this rodent
species in stables for heavy milking cows by testing preparations with active
substances of various origin. It involved an environmentally friendly product based
on sodium selenite 0.1%, a cholecalciferol-based natural product 0.75%, as well as
anticoagulant rodenticides containing the active substances bromadiolone 0.005%
and brodifacoum 0.005%. These preparations were formulated as granules, plate
bait or grain bait. The environmentally friendly sodium selenite product achieved
76.2% efficacy in the first year of research, 70% in the second, and 67.5% in the
third. The synthetic products based on bromadiolone and brodifacoum
demonstrated high efficacy in all of the three experimental years and in all three
formulations. The cholecalciferol rodenticide had 71.4% efficacy in the first year,
68% in the second, and 67.7% in the third. The data show that the environmentally
safe product had a lower efficacy due to high rodent abundance and inadequate
epidemiological conditions existing on the farm of heavy milking cows, while the
bromadiolone and brodifacoum-based products achieved high efficacy.
Key words: Rattus norvegicus, sodium selenite, efficacy, farm of heavy
milking cows
Introduction
Brown rat (Rattus norvegicus) is one of the most important pests in a group
of small rodents, and it is associated with other wild animals while living in the
closest neighbourhood of man and his environment (Bojkovski et al. 1991). It is

624
S. Đedović et al.
almost invariably found on cattle farms, where it builds a simple system of
underground corridors and makes 6-10 cm holes in walls. Rats are transmitters of a
number of infectious diseases to domestic animals (Morita, 1985). The chain of
toxicoinfection on a farm includes the farmers’ food and clothes, infected by rat
urine and feces, as well as hay and silage feed for cattle and direct rat bites.
Contaminated or poorly stored forage is a health hazard for animals (Daniels et al.,
2003).
Singleton et al. (Singleton et al.,1999) presented the environmental benefits
of a strategy of brown rat control by setting up points of attraction, i.e. a system of
trap-barriers rich in different food sources. A research has shown that facilities
housing different categories of animals are highly suitable places for rats to spread
into from surrounding environments as food sources are abundant there. This
research focused on the control of brown rat (Rattus norvegicus) on a farm with a
free range system of herd raising by application of an environmentally friendly
product based on selenium and on comparing the results with the effectiveness of
anticoagulant products and a cholecalciferol-based natural product, considering the
aspect of animal hygiene.
The micronutrient selenium used in forage in the amount of 0.1 mg/kg is
the optimum rate that helps protect health and improve productive and reproductive
characteristics of dairy cows. Calves fed after birth on milk supplemented with
selenium are better protected from antioxidative substances (Joksimović-Todorović,
2007). The mechanism of activity of sodium selenite is based on a replacement of
the S-H group of functional cellular proteins with the S-S bond, toxicity depends
on the chemical form of selenium, and the selenite is more toxic than selenate.
Target organs include: respiratory organs, CNS, cardiovascular system, digestive
system and skin. Lethal effect is achieved by a single dose and resistence is not
possible.
Introduction of these products would help reduce excessive use of the
anticoagulant products bromadiolone and brodifacoum, as well as frequent
intoxications of humans and domestic animals.
Cholecalciferol-based (vitamin D) products belong to a group of fast-acting,
acute poisons, whose mode of activity is based on an extensive absorption of
intestinal calcium and intensified resorption from bones, which leads to calcium
and phosphorus sedimentation in all soft tissues and broodstream, and results in
heart, blood vessels and kidney damage. The process of calcification is very fast in
the group of mice rodents, lasting no more than several days (Merck Veterinary
Manuel, 2011). The primary role of cholecalciferol is efficient control of rodent
pest populations that are resistent to anticoagulants (Marshall, 1984).
Cholecalciferol use has been limited in Serbia (Službeni glasnik RS, 2009),
which has narrowed the choice of available rodenticides.
The second generation anticoagulant rodenticides based on bromadiolone
and brodifacoum are slow-acting poisons whose mechanism of activity is based on

Control of brown rat (Rattus norvegicus) on a dairy farm...
preventing the synthesis of factor IX required for blood clotting, which results in
animal bleeding to death because its blood can no longer coagulate. Resistence has
been observed and reported (Lund, 1984, Myllymaki, 1995).
This research focused on the control of brown rat (Rattus norvegicus) on a
farm with a free range system of herd raising by application of an environmentally
friendly product based on selenium and on comparing the results with the
effectiveness of anticoagulant products and a cholecalciferol-based natural product,
considering the aspect of animal hygiene.
Materials and Methods
A three-year research was conducted on a farm of heavy milking cows that
are held in a free range system, and trials were set up in all available facilities: cow
stables, milking parlour, calves’ stable and silo. Before setting up trials and chosing
rodenticides, we examined the level of invasiveness by rodents, their paths and
numbers. Brown rat (Rattus norvegicus) presence was confirmed visually. The
appearance of feces that were found in the facilities further confirmed its presence.
All rodenticides were tested in these trials in different formulations and
they have different mechanisms of activity. The products EKOSTOP, EKOSEL
and MAMAK B (all in three formulations: granules, peletes, soft bait) were
manufactured by DOO CIKLONIZACIJA, Primorska 76, Novi Sad, Serbia, while
the active ingredient brodifacoum and the product RATTACK were manufactured
by DOO DUOCHEM, Ruzveltova 36, Belgrade, Serbia.
The standard method PP 1/114(2) (EPPO, 1999) was applied in all trials.
All treatments were made in the same period to reduce risks from various timedependent
factors (such as weather conditions or the loading and unloading storage
goods). Poisons were laid after previously placing placebo baits to carry out
method of census. Portions of 200 g of grain per box were laid down placed on the
first day of census. Rodent abundance was assessed using the C-30 method (World
Health Organization) or the transect method (EPPO, 1992) at the beginning and 10
days after the beginning of trial. Placebo bait was laid at 10 spots in each facility
over a period of 5 days. Each box was marked by an ordinal number, name of
product and amount of bait and arranged at 3-5 m distance from each other. To
avoid unwanted poisoning, the test products were labelled according to HACCP
standards (Bokelman,1996). The amount of consumed bait was checked on a daily
basis over a period of ten days. Bait was added as needed.
Data on palatability and biological effectiveness of products under
laboratory conditions and rodenticide efficacy against rodents in storage facilities
were calculated using Abbott's formula (Abbott,1925).
625

626
Results and Discussion
S. Đedović et al.
Brown rat was found in high abundance (Tables 1, 2, 3) on the dairy farm
as a result of its technology of construction, external unprotectedness and
inadequate sanitary and epidemiological conditions.
Figure 1 shows a ten-day intake of products based on different active
ingredients and formulated as granules (GB) in the first year of research.
bait intake
(g)
400
350
300
250
200
150
100
50
0
0 1 2 3 4 5 6 7 8 9 10
days
Sodium-selenite
Cholecalciferol
Bromadiolone
Brodifakum
Figure 1. Ten-day intake of products containing different active ingredients in granule (GB)
formulations on a dairy farm in the first year of research
The
granule formulation of all products was tested in the first year of
research.
The sodium selenite product was less palatable due to its relatively poor
organoleptic properties. Its palatability was highest in the first two days, with 280 g
eaten on the first and 290 g on the second day. Over the next several days, the
amount of bait consumed continued to decrease. The product should be improved
in terms of attractiveness, especially for situations in which animals have a wide
selection of food sources on a farm. High efficacy of this product was observed but
the experimental data reflected the known fact that selenium is highly degradable.
The fast-acting cholecalciferol was found to have been consumed the least,
which may be due to the fact that vitamin D concentration in a bait and its
palatability are inversely proportionate. Meehana (Meehan, 1984) had observed

Control of brown rat (Rattus norvegicus) on a dairy farm...
that a 0.1% concentration of cholecalciferol had a repelent effect on rodents. The
highest daily intake was observed on the third day after the trial started and it
decreased drastically and continually throughout the trial. Symptoms of poisoning
occurred 24-48 h after intake.
Palatability of this product should also be improved, especially on farms
where other food sources are also available.
The slow-acting rodenticides based on bromatiolone (0.005%) and
brodifacoum (0.005%) demonstrated good palatability.
The most palatable product was bromadiolone in this
year of research after ten
days of consumption, and its efficacy was as high as 94.8% (Table 1). Intakes of
both products decreased after seven days, which is consistent with out earlier data.
Mortality had a peak between the fourth and seventh day of consumption.
Figure 2 shows a ten-day intake of products containing different
active
ingredients formulated as plate bait (PB) in the second year of experiment.
bait intake
(g)
400
350
300
250
200
150
100
50
0
0 1 2 3 4 5 6 7 8 9 10
days
Figure 2. Ten-day intake of products containing different active ingredients in plate bait (PB)
formulations on the dairy farm in the second year of research
Plate baits were tested in the second year of research. Brown rats
consumed considerably greater amounts of sodium-selenite bait (0.1%) and product
palatability was satisfactory for the animals’ organoleptic requirements. Efficacy
627
Sodium-selenite
Cholecalciferol
Bromadiolone
Brodifakum

628
S. Đedović et al.
was slightly lower (70%) than it had been in the first year, which may be due to
degradation of the active ingredient.
The intake of bait based on cholecalciferol (0.075%) was lowest, only 936 g.
The ten-day consumption was even, slightly higher initially and then decreasing.
Palatability was unsatisfactory and the product achieved lower efficacy than it had
been observed in our earlier research. Bitter taste of this active ingredient makes
the animals give it up easily and exchange it for other available food on the farm.
Slow-acting rodenticides based on bromadiolone (0.005%) and brodifacoum
(0.005%) demonstrated better palatability in the second year, which may be due to
a better choice of attractant for rats and the fact that bait was coated with fine
transparent paper which causes curiosity in animals. We also found scattered bait
filter bags around rat paths together with dead animals. Total intake of
bromadiolone bait was 1667 g, and of brodifacoum 1068 g. However, the former
bait again had lower efficacy than brodifacoum even though its amounts consumed
were higher.
Figure 3 shows a ten-day intake of products with different active ingredients in
grain bait (AB) formulations in the third year of experiment.
bait intake
(g)
400
350
300
250
200
150
100
50
0
0 1 2 3 4 5 6 7 8 9 10
days
Sodium-selenite
Cholecalciferol
Bromadiolone
Brodifakum
Figure 3. Ten-day intake of products containing different active ingredients in grain bait (AB)
formulations on the dairy farm in the third year of research
Grain bait formulation was tested in the third year of research. Palatability
of the sodium-selenite and cholecalciferol products was equal in the initial three
days of trial. Their afficacies were also matching, 67.5% and 67.7%, respectively

Control of brown rat (Rattus norvegicus) on a dairy farm...
(Table 3). Total intake of the environmentally safe bait with sodium selenite was
1251 g and of the cholecalciferol product 1200 g. More animals died by eating bait
that contained sodium-selenite than cholecalciferol bait. It confirms our earlier
findings that sodium-selenite is highly toxic, and our future research will be
focused on improving its organoleptic properties to increase attractiveness.
Regarding efficacy and palatability of the anticoagulant products, data in the
third year of experiment confirmed consistency with those for the previous two
years, 95% efficacy was achieved by bromadiolone, and 100% by brodifacoum
(Table 3). The anticoagulant brodifacoum had the best attractiveness, good
palatability and the best efficacy throughout the three-year period of research. An
explanation of this may be the fact that its use had been limited in our country in
the past, so that resistance has not yet developed. Its palatability is especially
favourable. Palatability of the bromadiolone product was better in granules and
plate bait (Figures 1 and 2) than in grain bait (Figure 3).
Tables 1, 2 and 3 show efficacy data for the tested products with their different
active ingredients and different formulations.
Table 1. Product efficacy in the first year of research and estimated brown rat number
Species
Rattus
norvegicus
Active ingredient Bait intake Estimated rat number Efficacy
(product formulation) (g) Beginning End %
Sodium selenite (GB) 1251 42 10 76.2
Cholecalciferol(GB) 1200 28 8 71.4
Bromadiolone (GB) 1699 57 3 94.8
Brodifacoum (GB) 1242.5 43 0 100
Table 2. Product efficacy in the second year of research and estimated brown rat number
Species
Rattus
norvegicus
Active ingredient Bait intake Estimated rat number Efficacy
(product formulation) (g) Beginning End %
Sodium selenite (PB) 1438 50 15 70
Cholecalciferol(PB) 936 25 8 68
Bromadiolone (PB) 1667 55 3 94.5
Brodifacoum (PB) 1068.5 38 0 100
Table 3. Product efficacy in the third year of research and estimated brown rat number
Species
Active ingredient
(product formulation)
Bait intake
(g)
Estimated rat number
Beginning End
Efficacy
%
Sodium selenite (AB) 1245 40 13 67.5
Rattus Cholecalciferol (AB) 1065 31 10 67.7
norvegicus Bromadiolone (AB) 1235 40 2 95
Brodifacoum (AB) 1115 39 0 100
629

630
S. Đedović et al.
The environmentally friendly product based on sodium-selenite (0.1%)
demonstrated a satisfactory efficacy. It was 76.2% in the first year, 70% in the
second and 67.5% in the third (Tables 1, 2, 3). Better efficacy results could be
achieved by selecting the most viable attractant. Our own research so far has shown
high toxicity of this active ingredient, but little effort has been made so far to
improve its attractiveness and stability.
The cholecalciferol product was found to be less effective in its three
examined formulations in the three years of this researchthan it had been recorded
in our previous studies, namely 83-89% (Vukša et al., 2006., Đedović et al., 2011),
and in reports by other authors (Quy et al., 1995). The data possibly reflect the fact
that this active ingredient has a bitter taste which is unacceptible to the brown rat.
In addition to that, the farm had other food sources.
Bromadiolone granules and plate baits were found to have greater
palatability (Figures 1 and 2) than the product’s grain bait formulation (Figure 3),
but all formulations achieved an efficacy of around 95%, which indicates that
cumulative effects of this active ingredient had the same level of toxicity over a
period of 4-7 days (possibly because resistence has not developed). Rowe et al.
(1981) had reported bromadiolone efficacy ranging from 92.7-100% against the
house mouse on farms. Experiments so far and our overall experience have
indicated an extended activity of this active ingredient and animals were dying
even 10 days after treatment. Lower efficacy of these products is indicative of
resistance.
In this research, the rodenticides based on brodifacoum achieved 100%
efficacy (Tables 1, 2 and 3). Attractiveness was good in all three years of
experiment.
Conclusion
In our research, the sodium-selenite products had lower palatability, so that
suitable attractants need to be added to improve its organoleptic properties, and
yellow or green colour should be used because rats are particularly sensitive to
them. These products are highly toxic and laboratory research has confirmed it.
Cholecalciferol rodenticides were consumed most reluctantly due to their
bitter taste. They belong to a group of fast-acting poisons that have lethal effect
within 24 hours, but their negative side is that they have to be kept at low
temperature or fast degradation will occur. Our research has shown them as the
least effective.
The two anticoagulant products demonstrated good palatability and efficacy
in all tested formulations. Those are slow-acting poisons that have cumulative
effect which requires some time after intake. They should be carefully used
because of possible resistance.

Control of brown rat (Rattus norvegicus) on a dairy farm...
In this three-year research, product formulation was not found to have significantly
affected palatability for the brown rat.
We recommend regular three-month checks on farms with particularly high
infestations, poor hygiene and epidemiological conditions, and six-month checks
on farms with satisfactory conditions. Also, population abundance of brown rat
should be monitored on a monthly basis by setting up live-traps and marking
captured animals.
Acknowledgment
This study was carried out as part of Project III 4608 ”Development of an
integrated system of management of harmful organisms on plants, aiming to
overcome resistance and improve food quality and safety (2011-2014)“ which was
financially supported by the Ministry of Education, Science and Technological
Development of the Republic Serbia.
Prilog poznavanju suzbijanja sivog pacova (Rattus
norvegicus) na farmi mlečnih krava
S. Đedovic, M. Vukša , M. M. Petrović, J. Bojkovski, I. Pavlović, G. Jokić, B.
Stojnić
Rezime
Rattus norvegicus gotovo uvek živi u objektima namenjenim za gajenje
domaćih životinja i kao sinanotropna vrsta prisutan je u čovekovoj najbližoj
okolini. Predmet naših trogodišnjih istraživanja je alternativa smanjenja
ekonomskih šteta koju pričinjava ovaj glodar u stajama farmi visoko-mlečnih
krava, primenom preparata na bazi aktivnih materija različitog porekla.
U eksperimentima je korišćen ekološko prihvatljivi preparat na bazi
natrijum-selenita (0,1%), preparat prirodnog porekla na bazi holekalciferola
(0,75%) i antikoagulantni rodenticidi na bazi bromadiolona (0,005%) i
brodifakuma (0,005%).
Primenjeni preparati su formulisani u obliku granula, obloženog i zrnastog
mamka.
Ekološko prihvatljivi preparat na bazi natrijum-selenita je ispoljio efikasnost
od 76,2% u prvoj godini istraživanja, u drugoj 70% i u trećoj godini 67,5%.
Sintetisani preparati na bazi bromadiolona i brodifakuma pokazali su visoku
efikasnost u sve tri godine ispitivanja za sve tri navedene formulacije. Rodenticid
na bazi holekalciferola je ispoljio efikasnost od 71,4% u prvoj godini, u drugoj
631

632
S. Đedović et al.
68% i u trećoj 67,7 %. Rezultati istraživanja ukazuju da je zbog velike brojnosti
glodara i neadekvatnih epidemioloških uslova na farmi visoko-mlečnih krava
ispoljena slabija efikasnost ekološko prihvatljivog preparata dok su preparati na
bazi bromadiolona i brodifakuma ispoljili visoku efikasnost.
References
ABBOTT W.S. (1925): A method of computing the effectiveness of an
insecticide. J. Econ, Entomol, 18, 265-267.
BOJKOVSKI J., SOLDATOVIĆ B., NIKOLIĆ A. (1991): Cytogenetic
observation on numerical and structural chromosome aberrations induced by
environmental influence in mice of the genus Apodemus, Acta Veterinaria, 41, 5-
6, 241-246.
BOKELMAN B. (1996): Quality Assurance and HACCP with Special Attention to
Long Life Products.
DANIELS M.J., HUTSCHINGS M.R., GREIG A. (2003): „The risk of disease to
livestock posed by contamination of farm stored feed by wildlife excreta“.
Michigan Bovine Tuberculosis Bibliography and Datubase. Paper 14.
ĐEDOVIĆ S., VUKŠA M., STOJNIĆ B., JOKIĆ G. (2011): Preparat na bazi
selena i suzbijanje sinantropnih glodara u objektima u poljoprivredi. Pestic.
Fitommed., 26, 3, 265-270.
EPPO (1992):Guide for the officacy evaluation of rodenticides, No. 169, EPPO
Bull.,22,181-202.
EPPO, 1999: Guideline for the efficacy evaluation of rodenticides: Field tests
against synanthropic rodents (Mus musculus, Rattus norvegicus i R. rattus) – PP
1/114(2), in EPPO 1999. Guidelines for the efficacy evaluation of Plant Protection
Products. Rodenticides, 1, European and Mediterranean Plant Protection
Organization, Paris, pp. 83-144.
JOKSIMOVIĆ-TODOROVIĆ M., DAVIDOVIĆ V., HRISTOV S., STANKOVIĆ
B., BOJKOVSKI J. (2007): Deficit selena kod mlečnih krava., Zbornik naučnih
radova Instituta PKB Agroekonomik, 13, 3-4, 41-46.
LUND M. (1984): Resistance to the second generation anticoagulant rodenticides.
In: Proceedings of Eleventh Vertebrate Pest Conference. Sacramento, California
(Ed. by D.O.Clarck), pp. 89-94.
MARSHALL E.F. (1984): Cholecalciferol: a unique toxicant for rodent control.
Proc. 11 th Vertebr. Pest Conf., 95-98.
MERCK VETERINARY MANUEL (2011): Rodenticide Poisoning Introduction,
Whitehouse Station, NJ USA.
MEEHAN A.P. (1984): Rats and Mice-their biology and control. Rentokil Ltd.,
East Grinstead, W. Sussex, 383 pp.

Control of brown rat (Rattus norvegicus) on a dairy farm...
MORITA C.H., MATSUURA Y., MORIKAWA S.H., KITAMURA T (1985):
Age dependent transmission of hemoragic fever with renal syndrome (HFRS) virus
in rats. Arch. Virol., 85, 145.
MYLLYMAKI A. (1995): Anticoagulant resistance in Europe: Appraisal of the
data from 1992 EPPO questionnaire. Pestic. Sci., 43, 69-72.
ROWE F.P., PLANT C.J., BRADFIELD A. (1981): Trials of the anticoagulant
rodenticides bromadiolone and difenacoum against the house mouse (Mus
musculus L.). Journal of Hygiene, London 81, 171-177.
QUY R.J., COWAN D.P., PRESCOTT C.V., GILL E.J., KERINS G.M.,
DUNSFORT, G., JONES A., MACNICOL A.D. (1995): Control of a
Population of Norway Rats Resistant to Anticoagulant Rodenticides. Pestic.
Sci., 45, 247-256.
SINGLETON G.R., HINDS L.A., LEIRS H., ZHANG Z. (1999): Ecologically
based rodent menagment ACIAR, Canberra, Australia..
SLUŽBENI GLASNIK RS BR. 36/09 (2009): Lista zabranjenih hemikalija,
pravilnik o uvozu i izvozu određenih opasnih hemikalija.
VUKŠA M., DRAGANIĆ M., ĐEDOVIĆ S., JOKIĆ G. (2006): Laboratory
effects and efficacy of a Se-based rodenticide in controlling rodents in storage
facilites. Proceedings of the 9 th International Working Conference on Stored
Product Protection, Campinas, Sao Paulo, Brazil, 920-925.
Received 25 July 2012; accepted for publication 17 September 2012
633

Instruction for authors
Papers for publishing in the Biotechnology in Animal Husbandry journal
should be submitted to the Editorial Board. Address: Institute for Animal
Husbandry, Autoput 16, 11080 Belgrade-Zemun, P.O.box 23, Republic of Serbia
(for Biotechnology in Animal Husbandry).
Original papers in English, (on a CD-ROM or by e-mail:
biotechnology.izs@gmail.com) 6 pages of typed text using, Paper size: Custom
size, Width 17 cm, Height 24 cm; formata (Portrait), normal spacing (Single
Space). Margine: Top 2,0 cm, Left 2.0 cm, Bottom 2.0 cm, Right 2,0 cm, no
pagination. Use font Times New Roman, size 11 (except where it is stated
otherwise). Title of the paper should be Times New Roman, font size 14, bold:
Example 1
TABLE EGGS OF KNOWN ORIGIN AND GUARANTEED
QUALITY - BRAND EGG
Authors, Times New Roman, font size 12, bold
Z. Pavlovski, Z. Škrbić, M. Lukić
Institute for Animal Husbandry, Autoput 16, P. Box 23, 11080, Belgrade-Zemun, Republic of Serbia
Corresponding author: zlaticapav@yahoo.com
Invited paper
Example 2
THE EFFECT OF PARAGENETIC FACTORS ON
REPRODUCTIVE TRAITS OF SIMMENTAL COWS
M. D. Petrović 1 , Z. Skalicki 2 , V. Bogdanović 2 , M. M. Petrović 3
1 Faculty of Agronomy, Cara Dušana 34, 32000, Čačak, Republic of Serbia
2 Faculty of Agriculture, Nemanjina 6, 11080, Belgrade-Zemun, Republic of Serbia
3 Institute for Animal Husbandry, Autoput 16, P. Box 23, 11080, Belgrade-Zemun, Republic of Serbia
Corresponding author: petrovicm@tfc.kg.ac.yu
Original scientific paper
use 1,2, ... numbers in suffix to refer to addresses of authors, under affilations of
authors should be mentioned e-mail of corresponding author and category of paper,
Times New Roman, font size 9
Original scientific paper should contain following paragraphs with single spacing
(title of paragraphs should be in Times New Roman 14 bold, except for Abstract
and Key words where font size is 11 bold):
Abstract: 250 words

Key words: state key words (not more than 6)
Introduction - present the review of previous research and objective of the
paper.
Materials and Methods - state methods applied in the paper.
Results and Discussion - present investigation results separately from
discussion or together in one paragraph. Presentation of the results should be
precise and without repetitions, and include the evaluation of significant
differences and other parameters.
Text and titles of tables, figures and graphs, Times New Roman, font size 9, bold,
in the following form:
Table 1. Least square means for the reproductive traits of cows
Tables and figures should be numbered and with adequate title and legend, width
and height not exceeding 12 cm and 17 cm, respectively. Tables should be
prepared according to instruction for forming of tables in Office Word. Each
column in table must have heading and, when necessary, abbreviations should be
explained in the legend/footnote.
Conclusion - containing the most important issues of the paper
Acknowledgment - for example:
Research was financed by the Ministry of Science and Technological
Development, Republic of Serbia, project TR 6885.
After Acknowledgment the title of the paper in Serbian in Times New Roman 14
bold, is stated, followed by authors in Times New Roman 11 italic, example:
Konzumna jaja poznatog porekla i garantovanog kvaliteta -
brand jaja
Z. Pavlovski, Z. Škrbić, M. Lukić
Summary - should contain the most important issues of the paper. It should be
in English, and Serbian for domestic authors (min. 250 words).
References - should be in alphabetical order. Names of the authors must be
given in capital letters followed by the year of publicationin brackerts, titles in the
language of the original, examples:
PAVLOVSKI Z. (2004): Novi propisi EU, dobrobit živine, zahtevi potrošača.
Živinarstvo, 8-9, 49-58.
PAVLOVSKI Z., MAŠIĆ B. (1994): Odnos potrošača prema živinskim
proizvodima. Živinarstvo, 7-9, 77-82.

PETROVIĆ D.M., GUTIĆ M., BOGOSAVLJEVIĆ-BOŠKOVIĆ S. (2004): Masa
teladi pri rođenju i njena varijabilnost kod krava simentalske rase. Agroznanje, 5,
1, 111-116.
Citations in the text are presented in italic form, examples: ...results of Pavlovski
(2004)...; (Pavlovski and Mašić, 1994); (Petrović et al., 2004); (Pavlovski, 2004;
Pavlovski and Mašić, 1994; Petrović et al., 2004).
Authors are fully responsible for the contents of the papers.
Biotechnology in Animal Husbandry contains three categories of papers:
- Original scientific paper,
- Review paper, and
- Communication.
Review papers must have minimum 5 self-citations (by the first author).
All papers are published in English, and reviewed.
Abbreviation for journal Biotechnology in Animal Husbandry is:
Biotechnol Anim Husb
Editorial Staff

Institute for Animal Husbandry, Belgrade-Zemun
10th International Symposium
“Modern Trends in Livestock Production ”
2 nd – 4 th October 2013, Belgrade, Republic of Serbia
Esteemed colleagues,
FIRST ANNOUNCEMENT
Institute for Animal Husbandry, Belgrade-Zemun is organizing traditional
International Symposium, “ Modern Trends in Livestock Production ”.
Symposium will include following sections:
1. System of breeding of domestic animals
- Genetics
- Reproduction
- breeding
- selection
- nutrition
2. Production technology and quality of products
3. Animal welfare and health care
4. Livestock feed and ecology
5. Alternative production methods in livestock production
The work of sections will be divided per animal species.
A complete program of Symposium will be forwarded subsequently. For all
interested researchers kindly submit your papers in form of abstract (according to
instructions for authors-contributors of the journal "Biotechnology in Animal
Husbandry" - www.istocar.bg.ac.rs) before January 31 st 2013. A full paper
should be submitted by May 31 st 2013.
«Biotechnology in Animal Husbandry»
International Scientific Committee of the Symposium will review all submitted
papers.
The Committee will select papers for oral presentations. Remaining papers will be
presented in poster sessions. All papers presented on the Symposium will be
published in journal “Biotechnology in Animal Husbandry”.

Registration fee
• Registration fee which includes: publishing of paper in journal
Biotechnology in Animal Husbandry, Symposium material, participation in
all sessions of the Symposium, coffe/tea break, is 100 EUR (for domestic
participants in dinar value on the day of payment according to the
exchange rate). Papers will not be published without the payment of
registration fee.
• Registration fee which includes: publishing of paper in journal
Biotechnology in Animal Husbandry, Symposium material, participation in
all sessions of the Symposium, coffe/tea break, tourist programme and gala
dinner, is 150 EUR (for domestic participants in dinar value on the day of
payment according to the exchange rate).
Chairman Chairman
Organizing Committee International Scientific Committee
Dr Miloš Lukić, research associate Prof. Dr. Martin Wähner,
Serbia Germany
Please send abstracts and papers to following e-mail address:
Biotechnology.IZS@gmail.com
or by postal address:
Institute for Animal Husbandry
Organizing Committee
10th International Symposium
“ Modern Trends in Livestock Production ”
2 nd – 4 th October 2013, Belgrade, Republic of Serbia
Auto-put 16, 11080 Belgrade-Zemun, Republic of Serbia
phone. 011/2691-611; fax 011/2670-164

Institut za stočarstvo, Beograd-Zemun
10. međunarodni simpozijum
“Savremeni trendovi u stočarstvu ”
2. - 4. oktobar 2013. godine, Beograd, Republika Srbija
PRVO OBAVEŠTENJE
Institut za stočarstvo, Beograd-Zemun organizuje tradicionalni međunarodni
simpozijum, “Savremeni trendovi u stočarstvu ” sa sledećim temama:
1. Sistem gajenja domaćih životinja
- genetika
- reprodukcija
- gajenje
- selekcija
- ishrana
2. Tehnologija proizvodnje i kvalitet proizvoda
3. Dobrobit domaćih životinja i zdravstvena zaštita
4. Stočna hrana i ekologija
5. Alternativna proizvodnja u stočarstvu
Rad po sekcijama odvijaće se prema vrstama domaćih životinja.
Kompletan program simpozijuma dostavićemo naknadno. Molimo zainteresovane
autore da prijave svoje radove u obliku apstrakta (prema uputstvu za saradnike
časopisa „Biotechnology in Animal Husbandry“ – www.istocar.bg.ac.rs) i to do 31.
januara 2013. godine. Radovi u celosti se predaju do 31. maja 2013. godine.
«Biotechnology in Animal Husbandry»
Međunarodni naučni komitet, će nakon prijema radova izvršiti njihovu recenziju i
odabrati radove za usmenu prezentaciju. Ostali radovi će biti prezentovani u poster
sekcijama. Svi radovi sa Simpozijuma će biti štampani u celosti u časopisu
«Biotechnology in Animal Husbandry»

Kotizacija
• Kotizacija koja uključuje: štampanje rada u časopisu Biotechnology in
Animal Husbandry, materijal Simpozijuma, učestvovanje u svim sekcijama
Simpozijuma, kafu/čaj u pauzama, iznosi 100 EUR (za domaće učesnike u
dinarima na dan uplate po važećem srednjem kursu). Nijedan rad neće biti
štampan bez uplaćene kotizacije.
• Kotizacija koja uključuje: štampanje rada u časopisu Biotechnology in
Animal Husbandry, materijal Simpozijuma, učestvovanje u svim sekcijama
Simpozijuma, kafu/čaj u pauzama, turistički program i svečanu večeru,
iznosi 150 EUR.
Predsednik Predsednik
Organizacionog odbora Međunarodnog naučnog komiteta
Dr Miloš Lukić, Dr Martin Wähner,
Srbija Nemačka
Apstrakti i radovi u celosti se šalju na e-mail adresu:
Biotechnology.IZS@gmail.com
ili poštom na adresu:
Institut za stočarstvo
Organizacioni odbor
10. međunarodni simpozijum
“Savremeni trendovi u stočarstvu ”
2.- 4. oktobar 2013. godine, Beograd, Republika Srbija
Auto-put 16
11080 Beograd-Zemun
tel. 011/2691-611; faks 011/2670-164