P-139: Ram Sperm Motility Following 48 Hours of Refrigeration: Evaluation of Different Extenders for The Preservation of Ram Semen in Liquid State

Background: The application of laparoscopic method for intrauterine insemination is limited in sheep industry by the high cost of equipment and low pregnancy rate of frozen-thawed spermatozoa. Both disadvantages could be decreased if chilled ram semen is transported to the farms and inseminated through cervix. Semen extender provides the requirements for sperm and affects its motility, viability, and fertility during preservation in chilled condition. In this study we evaluated the motion characteristics of ram sperm which was diluted in egg yolk, milk, and soybean based extenders to find a suitable extender for chilled preservation of ram semen. Materials and Methods: Twenty ejaculates of 5 mature Zandi rams were collected once a week according to standard artificial insemination procedure. The samples were diluted in Tris-egg yolk, milked based (Inra; IMV France) and soybean based (Bioxcell; IMV France) extenders with concentration of 20 million/ml and preserved in refrigerator (4°C) for 48 hours. Sperm motion characteristics were evaluated in 1, 24, and 48 hours after refrigeration by Sperm class analyzer (Microptic, Barcelona Spain). Mean of data were compared using Duncan’s multiple range tests by significant level of 0.05. Results: Result showed that all of the kinetics parameters were reduced in time depend manner. The mean of motility were 88.17, 71.30 and 52.34 after 1, 24, and 48 of refrigeration respectively. Sperm motility, progressive motility and velocity patterns were significantly higher in Bioxcell extender at 1 and 24 hours compared than others groups. While after 48 hours of refrigeration milk based extender showed higher motility and velocity patterns compared with other two extenders. Egg yolk based extender indicated lower kinematic characteristics compared with others in all of the evaluated times. Conclusion: Our result showed milk based extender have more potential to preserved motion characteristics of chilled ram spermatozoa and were suitable to long term preservation of ram semen.