Effect of adding L-glutamine to Tris diluent on cryopreservation of coated sperm of Taleshi ram

Document Type : Research Paper


1 M.Sc. Student, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan

2 Associate professor, Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Rasht


Experiment was conducted to evaluate the effect of L-glutamine on coated sperm freezing by using four rams. Semen was collected by artificial vagina contact with a tube containing Tris-fructose-egg yolk 15%. Samples were pooled and centrifuged. After removing supernatant and diluting pellet, samples were split into 5 equal part and added 0, 40, 80, 120 and 160 mM L-glutamine. Samples were frozen by using liquid nitrogen After two weeks, samples were thawed and incubated at 37 °C for nine h. Sperm motility, plasma membrane integrity, viability (Hoechst 33258) and acrosome reaction (PNA-Alexa -flur-488) were investigated at 0, 3, 6 and 9 h. The main effect of L-Glutamine showed that sperm viability was higher in 0 (35.95%), 40 (40.65%) and 80 (45.95%) mM L-Glutamine than 160 mM L-Glutamine (30.40%; P<0.05). There was an interaction between incubation time and L-Glutamine on sperm motility and plasma membrane integrity (P<0.05). After thawing and 3 h later, the highest of plasma membrane integrity was observed in 80 mM L-Glutamine (49.25% and 47.34% respectively; P<0.05). Nine hours after thawing, the highest of sperm motility (26.4%) was observed in 80 mM L-Glutamine (P<0.05). Therefore, the addition of 80 mM L-glutamine to Tris-glycerol diluent (5%) improves coated sperm survival.


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