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.