Effect of Sperm Separation Methods on Morphology and Functions of Frozen Buffalo Spermatozoa

Abstract

This work was planned to compare three methods for selection of active buffalo spermatozoa, examine the effects of these separation methods on morphology, viability and functions of spermatozoa used for IVF purposes. Ten frozen straws per trial (10 times) were pooled and divided into 4 aliquots:  A) First aliquot was considered as control without any separation method. B) Second aliquot was subjected to sperm selection by density gradient method (percoll:PureSperm) using 40-80% double density gradient. C) The third aliquot was subjected to swim-up in sp-TALP. D) The fourth aliquot was subjected to washing by centrifugation with sp-TALP. The percentage of motility increased for Percoll, swim up and washing than control (86.0, 73.0, and 66.5 vs. 56.5) respectively. Sperm abnormalities % was significantly decreased after Percoll, swim up and sperm wash separation methods. Spermatozoa obtained by Swim up and Percoll had the highest percentage of intact membrane. Different spermatozoa separation methods significantly increased the lytic activity of the recovered spermatozoa. Live spermatozoa percentage with reacted acrosome significantly increased after both swim up separation and washing. The percentage of dead spermatozoa with reacted acrosome significantly decreased after percoll separation but it did not change when the swim up method was used. Finally it is concluded that, density gradient centrifugation using PureSperm® could be considered as the method of choice for selection of frozen thawed buffalo spermatozoa and presumably with a high potential fertilizing ability. density gradient centrifugation using PureSperm® could be considered as the method of choice for selection of frozen thawed buffalo spermatozoa.