4-EVALUATION OF MEMBRANE INTEGRITY OF BULL FROZEN-THAWED SPERM USING WATER AND HYPO OSMOTIC SWELLING TEST

Abstract

Assessment of the sperm membrane functional status appears to be a significantmarker for the fertilizing capacity of spermatozoa. The hypo osmotic swelling test(HOST) is one of the best methods to evaluate sperm membrane integrity. In thecurrent study, we used DW and hypo osmotic solutions of 50 and 100mOsm/l ofdextrose/NaCl, NaCl, sucrose and fructose. Based on the results, Among thedextrose/NaCl, NaCl, sucrose, and fructose solutions and DW, Maximum numbers ofswollen of bull frozen-thawed spermatozoa were observed with DW anddextrose/NaCl solution at 50 mOsm with average response by 61.20±8.677 and47.90±10.181 respectively. The HOST response at 3 and 60 min for all of solutionswere positively correlated to each other and there was no significant differencebetween the responses to the HOST at 3 and 60 min after incubation in all ofsolutions. The significant correlation was observed between motility anddextrose/NaCl at 50 mOsm, sucrose 50 and 100 mOsm, NaCl 50 mOsm and DW. Thehigh relationship was between motility and DW and dextrose/NaCl at 50 mosm. Therewas no significant correlation between DW and all of hypoosmotic solutions withstaining of the spermatozoa by eosin/nigrosin. In conclusion, the water test can beefficiently used for the evaluation of the functional integrity of the plasma membraneof bull frozen-thawed spermatozoa. The hypoosmular solution of dextrose/NaCl at 50mOsm is a good medium to evaluate bull frozen spermatozoa. The used of HOST andmotility are better tests to evaluate bull frozen thawed sperm than eosin-nigrosin. Theshort HOST procedure (3 min) is suitable method for evaluating of membraneintegrity of bull frozen/thawed spermatozoa.