Oral and Poster Abstracts

and durable spermatozoa. Within the same season (BS or Non BS)
significant differences are recorded in ejaculates of different motility
status (p0.001). This fact clearly suggests that the stability of
their cell membrane is rather a seasonal attribute than is related to
motility.
Key words: ram semen, semen kinematics, seasonality
888 Expression of Innate Immune Genes in Bovine Peripheral
Blood Leucocytes (PBL): Relevance to Endometritis in postpartum
Dairy Cows
A. Chapwanya 1 , K. Meade 1 , M. Doherty 2 , J. Callanan 2 , J. Mee 3 ,
C. O’Farrelly 1
1
Trinity College Dublin, School of Biochemistry and Immunology,
Dublin, Ireland
2
University College Dublin, School of Agriculture, Food Science and
Veterinary Medicine, Dublin, Ireland
3
Teagasc, Dairy Production, Fermoy, Ireland
Infectious bovine endometritis is the most common reproductive
disease of post-partum cows. In the immediate post-calving period,
all cows experience an influx of bacteria. Most cows will eliminate
all bacteria within 6 weeks of calving, but about 20% fail to do so
resulting in persistence and development of uterine disease.
However, little is known regarding the role of the uterine innate
immune response in dairy cows post-partum. We have recently
described 11 novel antimicrobial peptide (AMPs) genes in the
bovine genome. This study was designed to examine the expression
of these and other innate immune genes in PBL from post-partum
dairy cows. PBL were obtained from 12 dairy cows (healthy and
endometritic), RNA extracted and cDNA synthesised. Primers were
designed for selected genes involved in pathogen detection and
innate immune response to bacterial infections, including Toll-Like
Receptors (TLRs), cytokines and AMPs. The expression of these
genes was analysed by quantitative real-time PCR (qRT-PCR).
Preliminary results from 2 dairy cows show that known AMPs - LAP
and TAP are up-regulated in PBL from cows with endometritis.
BNBD4 expression remains unchanged in infection while the
expression of BNBD5 is reduced. Altered expression of innate
immune genes in bovine PBL may be a useful prognostic indicator of
uterine infection.
Key words: endometritis, AMPs, innate immunity, PBL
889 Susceptibility of ST, MDBK and BHK Cell Lines to Infection
with BoHV-1, bPIV3, BRSV and BVDV-1
R. Jordao 1 , C. Ribeiro 2 , E. De Stefano 2 , C. Del Fava 2 , C. Camargo 2 ,
E. Pituco 2
1
Instituto Biológico, Laboratório de Produçao de Imunobiológicos,
Sao Paulo, Brazil
2
Instituto Biológico, Centro de Pesquisa e Desenvolvimento de
Sanidade Animal, Sao Paulo, Brazil
Many viruses are potential contaminants of biological products
produced in bovine, porcine or hamster cells or manufactured via
processes using animal-derived raw materials such as bovine serum.
The susceptibility of cell lines was tested by inoculation with four
bovine viruses. Cell lines Swine Testicle (ST), Madin Darby Bovine
Kidney (MDBK) and Baby Hamster Kidney (BHK), were
experimentally infected with BoHV-1, bPIV-3, BRSV and BVDV-1
to determine if they were permissive for virus replication. To
determine whether cells were susceptible to viral infection, each cell
line was infected with one strain and incubated in 5% CO 2 at 37 °C,
until cytopathic effect (CPE) was seen, or harvested after four days
and were serially passaged until 32 days pos-infection (pi) with no
evidence of CPE, after this period when submitted by PCR and
ELISA for the presence of virus. ST cells showed evidence of CPE
200 XXV. Jubilee World Buiatrics Congress 2008
to BoHV-1 after 24 h (2nd passage - P2); to PI3 48h (1st passage -
P1); BRSV 72h (P1) and BVDV-1 48h (P2) pi. MDBK cells showed
evidence of CPE to BoHV-1 after 24 h (P1); to PI3 18h (P1); BRSV
96h (P1) and BVDV-1 48h (P1) pi. BHK cells showed evidence of
CPE to BoHV-1 after 48 h (P1); to PI3 48h (P1); BRSV 72h (P1) pi.
In contrast, cells do not showed evidence of BVDV-1 infection after
8th passage, ELISA assay and RT-PCR of experimentally infected
BHK cells demonstrated that there was no evidence of infectious
BVDV or contaminating viral genome. These results indicate that
the cells susceptibility of a species are not completely predictable
and that many cells derived from other species than cattle can be
infected with bovine virus, except BHK that is refractory to BVDV
infections. The presence of BVDV in cell cultures and in fetal bovine
serum has long been recognized problem not only in laboratories but
also among vaccine manufacturers. Consideration of the choice of
cell lines used in the production of vaccines also should be based on
the assessment of the risks of cross-contamination. In Brazil, a
vaccine for foot and mouth disease has been prepared in the BHK
cell line, and it is estimated that more than 395 million doses/year
have been administered to cattle. Viral contaminations are a serious
threat for animal cell cultures and may lead to false results in
research, development, and virus screening, to viral contaminations
in the biological products derived from the contaminated cultures.
Key words: cell culture, contaminants, susceptibility, viral contaminant
890 Effect of Mineral and Vitamin Treatment in Anovulatory
Anestrous Dairy Cows
M. Farhoodi, S. Mohamadi
Islamic Azad University, Science & Research Branch, Faculty of
Veterinary Medicine, Tehran, Iran
Introduction: The anestrous syndrome is a common and economically
important problem of world wide dairy farming. Anovulatory anestrous
is a multifactorial problem. Treatment options for anestrous cows are
different and include hormonal and management strategies and an
efficient treatment method of anestrous in dairy cows are preferred. The
objective of this study was to evaluate the effect of mineral and vitamin
therapy in anovulatory anestrous dairy cows.
Materials and Methods: The study was conducted in a large dairy
herd of 750 Holstein cows. All cows were examined for their
reproductive status 30 days postpartum. Cows, which had small
ovaries without palpable structures in rectal examination ten days
apart, were assigned as inactive ovaries anoestrous. These cows are
mostly in their first and second lactation. These anoestrous cows
were treated either with a single injection of GnRH (Buserelin) and
PGF 2a (cloprostenol) ten days later (GnRH group, n=89 cows) or
intramuscular phosphorus and Vitamins AD3E Therapy for three
days then injection of GnRH in third day and PGF 2a ten days later
(GnRH plus mineral-vitamin group n=58 cows). Reproduction data
were collected and signs of estrous (cure symptom) after each
treatment were recorded by visual observation (three times in day
heat detection) and confirmed by rectal palpation, examing uterus
and ovaries and checking mucous discharge. The curing rate was
calculated in each group and analyzed with chi-square test.
Results and Conclusions: In GnRH plus mineral-vitamin group 38
out of 58 cows (65.5%) showed estrous and in GnRH group 51 out of
89 cows (57.3%) showed estrous after treatment (p