THE PHILIPPINE WATER BUFFALO

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