THE PHILIPPINE WATER BUFFALO

the presence or absence of cysteamine. Intracytoplasmic
GSH values improved when oocytes were matured in
maturation conditions supplemented with cysteamine.
However, the values were lower compared to the GSH
levels of large follicles representing the in vivo matured
oocytes. The results of the present study suggest that an
increase in the intracellular GSH of bubaline oocytes
induced by cysteamine during maturation provide oocytes
with a favorable intracellular condition to support fertilization
and embryonic development in vitro.
Keywords: cysteamine, glutathione, MPN, cleaved
embryos
* Phil. J. Vet. Anim. Sci. 2002. 28(2):7-15. Also in
Proceedings of the 38 th Annual Convention, Philippine
Society of Animal Science, October 18-19, 2001, Manila,
Philippines. p. 1-2.
R-37
INTRACYTOPLASMIC GLUTATHIONE CONCEN-
TRATION OF SWAMP BUFFALO (Bubalus bubalis
limneticus) OOCYTES CULTURED IN VITRO*
L. C. Ocampo, Senior Science Research Specialist, PCC-CLSU, DA
M. B. Ocampo, Supervising Science Research Specialist, PCC-CLSU, DA
F. V. Mamuad, Center Director, PCC-CLSU, DA
E. V. Venturina, Laboratory Aide, PCC-CLSU, DA
T. Mori, Professor, Hokkaido University
H. Shimizu, Hokkaido University
Immature and maturing swamp buffalo (SB) COC’s were
cultured in vitro at different time points. At the end of each
time points, representative oocytes were denuded free of
cumulus cells and analyzed for their GSH content. The
GSH concentration of oocytes cultured at 0 h, 12 h, 18 h
and 24 h were 0.002 mm, 0.003 mm, 0.004 mm and
0.006 mm/oocyte, respectively. Similarly, the GSH
concentration of oocytes taken from preovulatory follicles
was 0.1853 mm/oocyte. The results suggest an increasing
pattern on the concentration of GSH as the maturation of
oocytes progresses. However, the concentration of
oocytes matured in vitro was lower compared to
preovulatory oocytes indicating possible defficiencies in
the culture medium used.
Keywords: swamp buffalo, glutathione, oocytes, in vitro
culture
*Buffalo Bulletin. 2000. 19(2):27-29.
R-38
MATURATION AND FERTILIZATION OF SWAMP
BUFFALO OOCYTES IN VITRO: EFFECT OF
LENGTH OF OVARIAN TRANSPORT*
REPRODUCTION
L. C. Ocampo, Senior Science Research Specialist, PCC-CLSU, DA
T. Mori, Professor, Hokkaido University
F. V. Mamuad, Center Director, PCC-CLSU, DA
M. B Ocampo, Supervising Science Research Specialist, PCC-CLSU, DA
H. Shimizu, Hokkaido University
The time during which the oocytes are left at postmortem
condition significantly affect the developmental competence
of oocytes. On this study, the timing of sequential changes
during 1 st meiotic division and the subsequent fertilization,
MPN formation and cleavage capability of swamp buffalo
oocytes aspirated at 2 hr and 6 hr post slaughter were
evaluated. In Expt. 1, oocytes recovered 2h and 6h after
slaughter, respectively, were cultured and fixed at 0, 6, 9,
12, 15, 18, 21, and 24 hr to evaluate meiotic progression.
Initial observation on the occurrence of the different meiotic
stages at each culture time point for oocytes recovered 6h
after slaughter indicated quite an advanced state of nuclear
progression compared to oocytes recovered 2h after
slaughter. In Expt. 2, oocytes were cultured for 24 hr
before in vitro fertilization. The resulting sperm penetration,
MPN formation and cleavage rate were higher even if not
significantly different when oocytes were aspirated 6h vs
2h after slaughter. This indicates that the time difference
utilized has not caused enough follicular changes to
affect the developmental competence of the oocytes.
Nevertheless, the results of the study show that swamp
buffalo oocytes acquire meiotic competence before in vitro
maturation. Therefore oocytes aspirated from ovaries at 2
hr to 6 hr postmortem condition can be used for the production
of early stage swamp buffalo embryos in vitro.
Keywords: swamp buffalo, maturation, fertilization,
ovarian transport
*Journal of Buffalo Science & Technique. 2001. 2: 68-77.
R-39
IN VITRO PRODUCTION OF PRE-IMPLANTATION
STAGE BUFFALO EMBRYOS*
M. B. Ocampo, Supervising Science Research Specialist, PCC-CLSU, DA
L.C. Cruz, Executive Director, PCC-OED, DA
F. V. Mamuad, Center Director, PCC-CLSU, DA
L.Ocampo, Senior Science Research Specialist, PCC-CLSU, DA
This study describes a technique for the in vitro production
of buffalo embryos from swamp buffalo (SB) oocytes
matured in vitro in condition with or without gonadotropin
addition. Higher proportion of SB oocytes was observed
to have undergone nuclear maturation in culture condition
with gonadotropin but provided no advantage in the overall
penetration, MPN formation and cleavage rate. Similarly,
the initial development early-stage embryos derived from
both treatment following co culture with cumulus cell showed
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