THE PHILIPPINE WATER BUFFALO
THE PHILIPPINE WATER BUFFALO
THE PHILIPPINE WATER BUFFALO
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priming prior to main FSH treatments improved further the<br />
ovulation rate in buffaloes.<br />
Keywords: water buffaloes, superovulatory, ovarian<br />
follicles, follicular dynamics<br />
*Philippine Journal of Veterinary Animal Science, 18 (1):19-32.<br />
R-11<br />
EMBRYO DEVELOPMENT RATE OF IN VITRO<br />
MATURED SWAMP <strong>BUFFALO</strong> OOCYTES<br />
CROSSFERTILIZED IN VITRO WITH RIVERINE<br />
<strong>BUFFALO</strong> SEMEN*<br />
D. H. Duran, Science Research Specialist II, PCC- CLSU, DA<br />
F. P. Aquino, Training Specialist III, PCC-CLSU, DA<br />
R. de Vera, Laboratory Aide, PCC-CLSU, DA<br />
L. C. Cruz, Executive Director, PCC-OED, DA<br />
A study was conducted to evaluate the in vitro development<br />
of F1 hybrid embryos which resulted from in vitro<br />
crossfertilization of in vitro mature swamp (2n=48, Philippine<br />
carabao) buffalo oocytes and riverine (2n=50, Murrah) buffalo<br />
semen. One hundred eighty three immature cumulus<br />
enclosed-oocytes from slaughter house derived swamp<br />
buffalo ovaries were cultured in TCM 199 suplemented with<br />
10% Fetal Bovine Serum (FBS) and antibiotics for<br />
maturation. After 23-24 hours of in vitro maturation (IVM),<br />
mature oocytes were partially freed from expanded cumulus<br />
cells and in vitro fertilization (IVF) commenced using frozenthawed<br />
semen from Murrah buffalo bull. After IVF, fertilized<br />
oocytes were further cultured for embryo development in the<br />
former maturation medium containing cumulus cells for 7<br />
days. Cleavage at 2-4, 5-6, and 7-8 cell stages<br />
approximately 24 hours after IVF were 39.34%, 10.93%<br />
and 3.28%, respectively, with total cleavage rate of 53.6%.<br />
Observed development rate of the cleaved embryos was<br />
10.20% morula, 15.31% blastocyst and 6.12% expanded<br />
blastocyst, and 6.12% hatched blastocyst on the 4 th , 5 th and<br />
6 th day after IVF, respectively. In swamp x swamp and<br />
river x river IVF, morula development in vitro was observed<br />
on the 6 th day of in vitro culture. These results suggest that<br />
heterosis effect may took place as early as the stage of<br />
zygotic gene activation among F 1 hybrid buffalo embryos.<br />
Keywords: in vitro maturation, in vitro fertilization, water<br />
buffalo oocytes<br />
*Proceedings of the PSAS 34 th Annual Convention. October<br />
23-24, 1997. Metro Manila, Philippines. p. 251.<br />
R-12<br />
EFFECT OF SOME PHYSICAL PARAMETERS ON<br />
EMBRYO DEVELOPMENT OF <strong>BUFFALO</strong> OOCYTES<br />
MATURED AND FERTILIZED IN VITRO*<br />
66<br />
ABSTRACT OF RESEARCHES ON<br />
<strong>THE</strong> <strong>PHILIPPINE</strong> <strong>WATER</strong> <strong>BUFFALO</strong><br />
D. H. Duran, Science Research Specialist II, PCC-<br />
CLSU, DA<br />
F. P. Aquino, Training Specialist III, PCC-CLSU, DA<br />
R. V. de Vera, Laboratory Aide, PCC-CLSU, DA<br />
P. Pedro, Senior Agriculturist, PCC-OED, DA<br />
L. C. Cruz, Executive Director, PCC-OED, DA<br />
Three studies were conducted to evaluate the effect of<br />
some physical parameters of embryo development of in<br />
vitro matured and in vitro fertilized buffalo oocytes. Study<br />
I was aimed to assess the effect of the type of culture dish.<br />
Two types of commercial dishes were compared; A =<br />
coated/treated (tissue culture dish) and B = non-coated<br />
(petri dish). To evaluate the brand effect of tissue culture<br />
dish, two brands, denoted as A 1 and A 2 were compared. A<br />
total of 229 cumulus-enclosed swamp buffalo oocytes were<br />
divided into three groups and cultured in vitro in the three<br />
dishes. Study 2 was designed to examine the effects of<br />
using aspiration medium during oocyte collection. Oocyte<br />
from 2-8 mm follicles were aspirated with or without<br />
aspiration medium and cultured for maturation and blastocyst<br />
development after in vitro fertilization. Study 3 was aimed<br />
to evaluate the effect of the types of oil that used to cover<br />
the culture medium. Three types of oil were compared;<br />
A=Mineral oil (marked embryo tested), B=Mineral oil (not<br />
marked) and C=Paraffin oil. Buffalo immature cumulus<br />
oocyte complexes were collected from slaughter housedderived<br />
ovaries, washed 3x and matured in TCM 199<br />
supplemented with 10% fetal bovine serum and antibiotics.<br />
In vitro fertilization was done using frozen-thawed semen<br />
capacitated with caffeine (5mM) and heparin (5ug/mM) in<br />
Bracket and Oliphant solution. Fertilized oocytes were<br />
cultured for 8 days in vitro using the same maturation<br />
medium with cumulus cell monolayer. Results in Study 1<br />
that cleavage rate in the tissue culture dishes A 1 (67.5%)<br />
and A 2 (60.3%) were significantly higher (P