THE PHILIPPINE WATER BUFFALO

differences at 16 to 32 cell stage only, but subsequently
development to the compacted morula-to blastocyst stage
provided no difference at all. The results presented here
indicate the possibility of using both maturation condition
and co culture system in the production of buffalo embryos
for basic research.
Keywords: buffalo embryos, gonadotropins, chick embryo
amniotic cavity
*Buffalo J. 2001. 17(1):145-159. Also in Proceedings of
the 34 th Annual Convention, Philippine Society of Animal
Science, October 23-24, 1997. PICC, Manila, Philippines.
p. 279-283. Also in Annotated Bibliography on Philippine
Biodiversity: Livestock and Poultry (Agrobiodiversity)
1949-1997, Carabaos. 2002. p. 397.
R-40
CLEAVAGE RATE OF BUBALINE OOCYTE MA-
TURED AND FERTILIZED IN VITRO*
M. B. Ocampo, Supervising Science Research Specialist, PCC-OED, DA
L. C. Ocampo, Senior Science Research Specialist, PCC-OED, DA
F. P. Aquino, Training Specialist, PCC-CLSU, DA
R. de Vera, Laboratory Aide, PCC-OED, DA
L. C. Cruz, Prof., DAS-CA, CLSU and Executive Director, OED-PCC-DA
The efficiency of modified systems for the in vitro maturation/
in vitro fertilization/ in vitro culture (IVM/IVF/IVC) of bubaline
follicular oocytes for the production of embryos was
determined. A total of 712 cumulus-oocyte complexes
(COCs) were selected for maturation and fertilization from
a pool of follicular fluid and oocytes that were collected by
aspiration from ovarian follicles. Of these, 351 oocytes
that were fixed 15 hour post insemination registering
maturation and fertilization rates of 82.3% and 57.8%,
respectively. The monospermic fertilization rate was
59.6%. The remaining 361 oocytes were allowed to
continue development for 30 hour obtaining a cleavage rate
of 29.9%. Of these, 78 and 30 embryos were cultured in
TCM–199 + 10% BCS + cumulus cell monolayer and
chick embryo amniotic (CEAm) fluid with a morula/
blastocysts formation rate of 23.1% and 36.7%,
respectively. These findings suggest that the IVM/IVF/
IVC systems adopted in this laboratory could be used for
the production of buffalo embryos in vitro.
Keywords: bubaline follicular oocytes, cleavage rate,
cumulus cell monolayer, in vitro matured/ in vitro fertilization/
in vitro culture
*Annual Report Embryo Technology Project, PCC. 1995.
Also in Abstracts of Water Buffalo Researches in the
Philippines 1981-1995. 1996. PCC-DA. p. 65. Also in
PCC R & D Highlights 1994-1996. 1997. p. 22-23. Also
78
ABSTRACT OF RESEARCHES ON
THE PHILIPPINE WATER BUFFALO
in Annotated Bibliography on Philippine Biodiversity:
Livestock and Poultry (Agrobiodiversity) 1949-1997,
Carabaos. 2002. p. 280.
R-41
IN VITRO MATURATION AND FERTILIZATION OF
WATER BUFFALO OOCYTES*
M. B. Ocampo, Supervising Science Research Specialist, PCC-CLSU, DA
L.C. Ocampo, Senior Science Research Specialist, PCC-CLSU, DA
F. P. Aquino, Training Specialist II, PCC-CLSU, DA
N. D. Lorenzo, Science Research Aide, PCRDC-PCARRD, CLSU
R. V. de Vera, Laboratory Aide, PCC-CLSU, DA
J. V. del Rosario, Staff, PCC-CLSU, DA
L. C. Cruz, Executive Director, PCC-OED, DA
Some factors such as oocytes selection, hormone
supplementation of the maturation medium, bull variability,
caffeine concentration, sperm concentration, and effect of
cumulus cells which influence the maturation, fertilization
and development of oocytes in vitro were investigated.
Higher concentration of oocytes with cumulus cells
completed the first meiotic division than when oocytes
were denuded. Supplementation of the maturation medium
with 10ug/ml of luteinizing hormone (LH) yielded higher
maturation rates than with lower (5ug.ml) LH
supplementation. Variability of the semen source affected
the penetration rates with MB #226 (among the 4 bulls
semen source) registering the highest fertilization rate. While
sperm exposure to different caffeine concentrations (2.5 to
10 mmol) had no effect on its penetrating capability,
increasing sperm concentration by 5 to 10 folds improved
the penetration rate but similarly lessened the rate of
monospermic fertilization. The co-culture of embryos
produced by IVF of in vitro mature buffalo oocytes with
cumulus cells resulted in blastocyst formation rate of 5%.
The co-culture of embryos may be a valuable tool in
identifying interactions between embryo and cells of the
reproductive tract, which plays a major role in providing
the necessary condition for normal embryonic development.
Keywords: water buffaloes, in vitro fertilization, in vitro
maturation, luteinizing hormone
*PCC R & D Highlights 1994-1996. 1997. PCC-OED,
DA. p. 22-23.
R-42
SOME FACTORS INFLUENCING THE MATURATION,
FERTILIZATION AND DEVELOPMENT OF BUFFALO
OOCYTES IN VITRO*
M. Ocampo, Supervising Science Research Specialist, PCC-OED, DA
L. Ocampo, Senior Science Research Specialist, PCC-OED, DA
F. Aquino, Training Specialist III, PCC-CLSU, DA