Comparative Efficacy of Percoll^TM Discontinuous Density Gradient Centrifugation and Glass Wool Filtration Techniques for Spermatozoa Selection in Buffalo (Bubalus bubalis)

Dead and damaged spermatozoa cells present universally in the ejaculates of all eutherian mammals exert toxic effect on contemporary healthy cells mostly through generation of excessive free radicals. This is much more evident during extended period of processing, resulting in poor ejaculate quality. The solution lies in depletion of dead/damaged spermatozoa from the neat ejaculates itself. Thus the objective of the study was to evaluate the efficiency of the protocols such as discontinuous Percoll^TM density gradient centrifugation (PDGC) and glass wool filtration (GWF) for depletion of dead/damaged spermatozoa from fresh semen in buffalo. Random ejaculates (n=6) of Murrah buffalo bulls were divided into two aliquots after quality assessment: PDGC and GWF protocols (Group I and II, respectively). At the end of the purification protocol, efficiency of the protocols in depleting dead/damaged spermatozoa as reflected by certain quality parameters were evaluated. The mean efficiency (%) of purification protocols based on recovery of spermatozoa was 44.68 and 40.02% for PDGC and GWF, respectively. Moreover significantly (p<0.05) greater values for quality parameters was observed in the Group II (26.4+6.8 vs 68.8+4.4 for acrosome integrity (%); 12.68+6.6 vs 57.7+7.5 for functional plasma membrane integrity (%); 20.3+5.8 vs 80.75+6.7 for viability (%) in Group I and II, respectively). It was concluded that GWF is a better technique than PGDC to filter out dead/damaged spermatozoa from fresh semen with improvement in semen quality and can be a valuable tool in assisted reproductive technology.