THE PHILIPPINE WATER BUFFALO

R. de Vera, Laboratory Aid, PCC-OED, DA
L. C. Cruz, Professor, DAS-CA, CLSU and Executive Director, PCC-OED, DA
Experiments were conducted to establish a workable in
vitro maturation, fertilization and culture (IVM/IVF/IVC)
techniques for buffalo oocytes. These include 1) selection
of oocytes, 2) hormone supplementation of maturation
medium, 3) bull variability, 4) caffeine concentration, 5)
sperm concentration and 6) co-culture of embryos cumulus
cells. Higher proportions of oocytes with cumulus cells
vs. denuded oocytes completed the 1 st meiotic division.
Similarly, LH, supplementation of the maturation medium
at 10ug/ml yielded a higher maturation rate than when LH
concentration was 5 ug/ml. When oocytes were fertilized
in vitro using semen samples from different bulls, variable
penetration rates were obtained. Bull No. 226 registered
the highest fertilization rate vs. Bull No. 39, 45 and 228.
Exposure of spermatozoa 9 from Bull No. 226 to different
levels of caffeine (e.g. 2.5 mM; 5 mM, 10mM) had no
significant effect on their penetrating capability. However,
increasing the sperm concentration at fertilization by 5-10
folds improved the penetration rate but similarly lessened
the rate of monospermic fertilization. Co-culturing of
embryos produced by IVF of IVM buffalo oocytes with
cumulus cells resulted to blastocyst formation rate of 5.0%.
Keywords: buffalo oocytes, in vitro maturation fertilization
and cultural techniques
*Annual Report Embryo Technology Project, PCC. 1995.
Also in Abstracts of Water Buffalo Researches in the
Philippines 1981-1995. 1996. PCC-D. p. 65. Also in
PCC R & D Highlights 1994-1996. 1997. p. 24. Also in
Annotated Bibliography on Philippine Biodiversity:
Livestock and Poultry (Agrobiodiversity) 1949-1997,
Carabaos. 2002. p. 282.
R-43
PREGNANCIES RESULTING FROM SWAMP
BUFFALO OOCYTES MATURED AND FERTILIZED
IN VITRO*
M. B. Ocampo, Supervising Science Research Specialist, PCC-OED, DA
L. C. Ocampo, Senior Science Research Specialist, PCC-OED, DA
N. D. Lorenzo, Science Research Specialist, PCC-CLSU, DA
F. P. Aquino, Training Specialist, PCC-CLSU, DA
R. V. de Vera, Laboratory Aid, PCC-OED, DA
L. C. Cruz, Professor, DAS-CA, CLSU and Executive Director, PCC-
OED, DA
Follicular oocytes (n=343) collected from buffalo cows at a
local abattoir were matured and fertilized in vitro. Of the 97
ova fixed for analysis, 67% were found presented with
58.5% monospermic fertilization rate. Of the 246 ova that
were allowed to continue embryonic development following
REPRODUCTION
insemination, 23.6% cleaved. These embryos were further
cultured in TCM 199 + cumulus cell monolayer or in the
amniotic fluid of a developing chick embryo (CEAm fluid)
for up to 6 and 5 days, respectively. Those embryos
selected at morula or blastocysts stage were transferred to
recipients. Two pregnancies resulted from the non-surgical
transfer of morula stage embryos. The results demonstrated
that immature buffalo oocytes can be matured and fertilized
in vitro, subsequently developed to morula or blastocysts
stage and develop into a normal pregnancy after transfer.
Keywords: buffalo oocytes, in vitro fertilization and
maturation
* Phil. J. Vet. Anim. Sci. 1997. 23(3 & 4):14-18. Also in
Annual Report. Embryo Technology Project, PCC. 1995.
Also in Abstracts of Water Buffalo Researches in the
Philippines 1981-1995. 1996. PCC-DA. p. 66. Also in
Annotated Bibliography on Philippine Biodiversity:
Livestock and Poultry (Agrobiodiversity) 1949-1997,
Carabaos. 2002. p. 281.
R-44
PRESENT STATUS OF EMBRYO TRANSFER IN
WATER BUFFALOES (A REVIEW)*
M. B. Ocampo, Supervising Science Research Specialist, PCC-CLSU, DA
L. C. Ocampo, Senior Science Research Specialist, PCC-CLSU, DA
A. A. Rayos, University Researcher, DTRI-CA, UPLB
H. Kanagawa, Professor and Chairman, DT-FVM, Hokkaido University
The great importance of the water buffalo as a major source
of draft power for farm tillage, transport and as a source of
supplementary income through milk and meat production
has triggered a keen interest in its reproduction. Techniques
to generate an economically feasible number of fertilized
eggs from one animal (donor), which eventually can be
transferred, to a number of surrogate mothers (recipients)
are being developed. Such techniques are referred to as
embryo transfer (ET) technology, which consists of several
steps, each of which is critical. These include: (a) selection
of donor and recipients, (b) superovulation, (c) estrus
synchronization between donor and recipients, (d) recovery
of embryos, (e) examination and classification of embryos,
and (f) transfer of embryos. Failure in any one of the
above steps would result in either a low conception rate or
complete failure. This technology has opened exciting
possibilities in enabling the introduction of breeding schemes
where nucleus breeding herd can provide an economical
method of interpolating the desired production traits. While
the application of this technology in the water buffalo requires
much additional research work, it has already been clearly
demonstrated that highly acceptable product can result.
The birth of a 35 kg male buffalo calf 300 days after non-
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