effect of different levels of l-glutamone and glycerol on coated ram spermatozoa in freezing procedure

to determine the effect of different levels of l-glutamine and glycerol on freezing coated sperm, semen were collected from 4 taleshi rams through artificial vagina contact with a tube containing tris-fructose-egg yolk 15%. ejaculates were pooled, centrifuged and supernatant was removed, then cell fractions were used. samples were split into 9 equal parts and 3 (g3), 5 (g5) or 7% (g7) glycerol and 0 (g0), 80 (g80) or 160 (g160) mm l-glutamine were added. then samples were frozen by using liquid nitrogen and thawed after 2 weeks. sperm motility, plasma membrane integrity, viability and acrosome reaction were investigated at 0, 3, 6 and 9 h after thawing. the results showed that there was an interaction between glycerol and l-glutamine on sperm motility and acrosome integrity. the highest sperm motility was observed in g80g5 (45±1.95), g0g7 (48.75±1.95) and g80g7 (50±1.95). the lowest sperm motility was achieved by g0g3 (26.5±1.95). the lowest acrosome integrity was observed in g160g3 (76±0.94). the highest plasma membrane integrity (45.28±1.57) and sperm viability (36.71±1.16) was observed in 80 mm l-glutamine. the highest sperm viability was observed in 7% glycerol (41.71±1.58). therefore, 7% of glycerol and 80 mm of l-glutamine could be beneficial to freeze coated ram spermatozoa.