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