Oral and Poster Abstracts

923 Production of Bovine Blackleg Vaccine by Fermenter
R. Pilehchian, M. Moosawi, A. Jabari
Razi Research Institute, Anaerobic Vaccine Production, Karaj, Iran
Blackleg is an actual and fatal disease of cattle caused by Clostridium
chauvouei. The object of this study was to prepare a potent vaccine
against blackleg of cattle. Attempts were made to prepare and formulate
the ingredients in order to obtain high yield of Clostridium chauvoei
organisms in culture medium by fermenter. The medium consisted of
efficient amount of nitrogen and carbon sources. A large-scale vaccine
with satisfactory results was obtained in fermenter. More, than
1,450,000,000 organisms/ml was obtained by the end of incubation
period. The safety and potency of the prepared vaccine was determined in
Guinea pigs and Sheep according to British pharmacopoeia (veterinary)
with satisfactory results. In a preliminary experimental trial, 9 calves were
divided into 3 groups. 2 groups were immunized by 2 doses of the
vaccine, with an interval of 15 days, and then challenged in 2 weeks after
the 2 nd injection. The third group was considered as a non-immunized
control. According to the results, the vaccine prepared an adequate
protection against the wild strain of Clostridium chauvouei. Animals of
the control group were rapidly died after injection of the challenge strain.
Key words: Bovine Blackleg vaccine, fermenter
924 Study on Production of Reduced Dose Blackleg Concentrated
Vaccine
M. Moosawi, R. Pilehchian, A. Jabari, L. Abdolmohamadi
Razi Research Institute, Anaerobic Vaccine Production, Karaj, Iran
Blackleg is generally known to affect cattle but sheep, goats, swine,
camels, deer and mink are also susceptible. Blackleg in cattle has been
recognized in Iran. Formalized whole culture vaccine have been
prepared for immunization of cattle and sheep against the disease at
Razi Institute using traditional manner and also by fermenter. The main
object of this study was to prepare and formulate a reduced dose potent
vaccine for immunization of cattle against blackleg in Iran. There are a
few references in the literature concerning production of blackleg
vaccine by fermenter. After an active growth duration, detoxification
and killing of bacteria was done by adding formaldehyde (37%) at the
rate of 0.7% and pH of culture adjusted to 7.Aluminum hydroxide gel as
adjuvant was used for the Clostridium chauvoei vaccine in sufficient
quantity to give 10 percent gel in final concentration. The concentrated
vaccine was mixed by slowly stirring for 30 minutes at 20 °C.The
quality control of the experimental vaccine was determined according to
the European pharmacoepia Veterinary 5 th edition (2004). Three healthy
cattle were selected for safety test of the vaccine. Each cattle was
injected subcutaneously 2ml of the vaccine and observed for ten days.
Ten healthy guinea pigs (300-400gr) were injected subcutaneously with
2ml of the vaccine as primary dose, followed by three weeks later a
second injection of the same quantity of the vaccine as the secondary
dose. After two weeks the vaccinated and unvaccinated guinea-pigs
were injected intramuscularly with 4 MLD for vaccinated and 1MLD
for unvaccinated of 24 hrs virulent culture of Clostridium chauvoei
respectively. Results shows that the routine blackleg vaccine with 3ml
dose gave good immunity in injected animals but the new experimental
concentrated vaccine with reduced dose of 2ml has given highly
immunity in guinea pigs and cattle. The reduced dose of vaccine could
be replaced of the routine blackleg vaccine because of long immunity in
vaccinated animals and also because of economic points of view.
Key words: Blackleg vaccine, cattle, fermenter, pharmacoepia
925 Bovine Embryo Development after In vitro Maturation with
Glycine
C. Silva, A. Martins Jr, R. Calegari
Sao Paulo State University - UNESP, Clinic, Surgery and Animal
Reproduction, Aracatuba SP, Brazil
The purpose of this study was to investigate the effects of glycine during
in vitro maturation of bovine oocytes on their subsequent embryonic
development after IVF. Oocytes were aspirated from follicles (2 to 6 mm)
of ovaries obtained from slaughtered cows. Oocytes having at least 3-4
layers of cumulus cells and homogeneous cytoplasm were washed in
TCM-199 with 2.2 mg/ml sodium bicarbonate, 0.02 mg/ml sodium
pyruvate, 0.05 mg/ml penicillin, 1 mg/ml PVA, and 2.38 mg/ml HEPES.
210 XXV. Jubilee World Buiatrics Congress 2008
Groups of 15 to 25 oocytes were matured for 24 h in 100 µl drops of TCM
(Gibco ® , USA) with 0.5 µg/ml FSH (Pluset ® , Spain), 50 Ïg/ml LH
(Lutropin-V ® , Canada), and 10% FCS (Nutricell, Brazil), supplemented
with glycine, according to the experimental groups: I (0,67mM; n=144),
II (10 mM; n=132), and III (100 mM; n=130). For inseminations, frozenthawed
sperm was obtained by density gradient separation (Percoll).
Following a centrifugation (700 x g) for 30 min, the resulting pellet was
washed in TALP by centrifuging 200 x g for 5 min. Afterwards, the pellet
was re-suspended to 50 µl with TALP plus PHE and heparin. Gametes
were co-incubated for 20 h for IVF. Then, cumulus cells were removed
and 15 to 20 presumptive zygotes were cultured in 50 µl drops of culture
medium (m-SOF) up to day 8 post-insemination (pi). Oocytes and
embryos were incubated at 39 °C under a humidified atmosphere of 5%
CO 2 in air. Embryonic development was monitored, and 50% of the
culture volume was replaced with fresh medium at 72 and 120 h pi.
Percentages of cleaved oocytes and oocytes reaching morula
(M)/blastocyst (B), blastocyst/expanded blastocyst (EB), and expanded
blastocyst/hatched blastocyst (HB) stages were recorded at 72, 144, 168,
and 192 h pi, respectively. Data of five replicates were analyzed using
ANOVA and Bonferroni t-test. Differences of P