P-119: In Vitro Assessment of Sucrose for Preservation of Buffalo Epididymal Spermatozoa

Background: Epididymal spermatozoa have been used for production of offspring from genetically important animals after they are dead. To attain optimal fertility with epididymal spermatozoa, they should be preserved in a suitable medium in 37°C. Preservation media provides requirement for spermatozoa and affects its motility, viability, and fertility. Sucrose is one of the important components of semen extenders. The beneficial effect of sucrose on preservation (cold and freeze condition) of semen has been examined in other species. While this has not been studied in buffalo epididymal spermatozoa. In this study we evaluate buffalo epididymal sperm parameters to find optimal concentration of sucrose in preservation media. Materials and Methods: Thirty testes from 15 mature buffalo bulls were collected from Urmia slaughterhouse and transported to laboratory in cool condition. Spermatozoa were obtained from caudal epididymis and pooled together. The samples were diluted in tissue culture media (TCM) at concentration of 20×106 sperm/ml and divided to five groups containing sucrose at concentration of 0, 2.5, 5, 7.5 and 10 mM. Sperm motion characteristics, viability, and cytoplsmic droplet were evaluated by computer assisted sperm analyzer and one steps eosin- nigrosin stain respectively at 1, 6, 12, and 24 hours of incubation at 37°C. Mean of data were compared using Duncan’s multiple range tests by significant level of 0.05. Results: The addition of sucrose in preservation media did not affect motility, viability, and velocity patterns of epididymal spermatozoa until 6 hours of incubation (p>0.05). While supplementation of sucrose at 5 and 7.5 mM significantly improved motility, viability, and curvilinear velocity of spermatozoa following 12 and 24 hours of incubation (p<0.05). Cytoplsmic droplets were reduced when 5 mM of sucrose was added in preservation media at 6 and 12 hours of incubation as compared to control group (p< 0.05). Conclusion: We conclude that addition of 5 mM sucrose in preservation media of epididymal buffalo spermatozoa results in the release of cytoplsmic droplets and improves motility and viability of spermatozoa during 24 hours preservation.