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Post mortem epididymal sperm (ES) is an excellent source of germplasm for conservation of animal genetic resources. In this study,  the viability of post mortem ES from carabao after collection at ambient temperature (AT) and after 24 hr and 48 hr of storage at refrigeration temperature (RT) were evaluated. ES were collected through slicing of epididymides from 7 carabaos. The mean ES volume was 0.4 ml with sperm concentration of 2.5×109 cells/ml. The mean percentage livability of fresh ES, after 24 hr and 48 hr of storage at RT were 81.93%, 65.93% and 43.7%, respectively. The mean percentage abnormalities of fresh ES, after 24 hr and 48 hr of storage at RT were 44.15%, 38.5% and 43.47%, respectively. The mean percentage motility of fresh ES, after 24 hr and 48 hr of storage at RT were 60.0%, 50.0% and 17.24%, respectively, after analysis through conventional means. Through CASA, the mean percentage motility was observed to be lower significantly at 26.12%, 37.94% and 18.32% for fresh ES, after 24 hr and 48 hr of storage at RT, respectively. The mean progressive motility of fresh ES, after 24 hr and 48 hr storage at RT were 10.02%, 16.47% and 10.87%, respectively. The results suggest that ES potential for use in fertilization studies remained viable when used immediately or after storage at RT for the first 24 hr.

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Buffalo ovaries (n=610) were collected from the abattoir and the oocytes recovered were matured, fertilized and the resulting zygotes were cultured in vitro. In Experiment 1, the maturation and fertilization rates of O 1 -O 2 oocytes were higher than O 3 -O 5 oocytes. In Experiment 2, the two batches of TCM-199 medium used with different hormonal supplementation did not influence the maturation and fertilization of the oocytes at 70.7-86.7% and 59.4-66.0% respectively. In Experiment 3, bull variability was observed in terms of their capacity for fertilization. In Experiment 4, higher fertilization rate was observed in response to caffeine (3-10 mg/ml) compared to control. In Experiment 5, 1 × 10 6 sperm/ml concentration during fertilization had a lower penetration rate (43.0%) than 5, 10 × 10 6 sperm/ml concentration (68.4-71.0%). In Experiment 6, development to the blastocyst stage in TCM-199 or mSOF medium with or without cumulus cell monolayer had no significant difference (11.5-22.2%) with total cell count ranging from 84.6±11.2 to 91.4±12.2. Overall, the results showed the importance of cumulus cells in the acquisition of developmental competence of oocytes maturing in vitro and its subsequent fertilization. That, selection of appropriate bull, its optimum sperm concentration, the choice of capacitating agent and its concentration, the type of medium for culture of resulting zygotes is critical for successful application of in vitro maturation, fertilization and culturing for mass production of buffalo embryos. Keywords: Embryos; fertilization; maturation; oocytes To cite this article: Ocampo LC, LM Rigos and MB Ocampo, 2015. Factors influencing the maturation, fertilization and development of swamp buffalo oocyte in vitro. Res. Opin. Anim. Vet. Sci., 6(1): 24-31.