Comparison of Motility, Morphology, Acrosome Integrity, Membrane Integrity and Fertilizing Ability of Frozen-Thawed Buffalo Sperm Separated by a Percoll

The aim of this study was to evaluate sperm quality following separation and selection using a silane-coated silica colloid method (PureSperm ® ) or Percoll ® . Frozen/thawed sperm from buffalo bulls (n=5) were assigned to two groups. The first group was separated using a Percoll ® discontinuous gradient (45 and 90%) and the second group by a single layer of PureSperm ® . Sperm motility, concentration, morphology, acrosome integrity and membrane integrity were evaluated and compared before and after sperm separation. Ability of sperm to fertilize was tested by hyaluronan binding assay (HBA) score. Sperm motility and morphology improved (P < 0.001) after the separation process with no differences between methods. The Percoll ® gradient increased the mean (± SEM) percentage of sperm recovered (8.27±0.79) compared to PureSperm ® (5.25 ± 1.41; P = 0.007). However, membrane integrity (61.4 ± 1.3 vs. 56.8 ± 1.3) and acrosome integrity (59.8 ± 1.4 vs. 55.3 ± 0.6) were higher following selection using PureSperm ® than Percoll ® (P = 0.03). The HBA score was higher when semen was selected by PureSperm ® than by Percoll ® (49.5 ± 1.6 vs. 40.0 ± 1.1; P = 0.02). We conclude that PureSperm ® separation resulted in lower sperm recovery than by Percoll ® but was more effective for selecting high quality sperm for in vitro fertilization for embryo production in buffalo.