SEPARATION OF MOTILE SPERMATOZOA FROM FROZEN-THAWED BUFFALO SEMEN - SWIM-UP VS FILTRATION PROCEDURES

Three experiments were conducted to maximize the recovery rate of moti le spermatozoa from frozen-thawed buffalo semen. In Experiment 1, the swim-up of motile spermatozoa was performed in the presence or absence of HEPES in TALP medium and CO2 in the environment. The recovery rate of motile spermatozoa in TALP medium (control), TALP+HEPES+CO2, TALPHEPES and TALP+CO2 was 15, 18, 12 and 10%, respectively (P>0.05), with sperm motility at 87 89, 90 and 90%, respectively (P>0.05). In Experi ment 2 the pH of TALP medium was adjusted to 7.0, 7.5, 8.0, 8.5 and 9. 0, and swim-up procedure was performed in the presence of HEPES and CO 2. The recovery rate of motile spermatozoa at different pH was 14, 20, 24, 27 and 16%, respectively (P<0.05). Motility of separated spermato zoa was 88, 91, 90, 89 and 90%, respectively (P>0.05). In Experiment 3 , the efficiency of ion-exchange filtration and Swim-up procedure in s eparating motile spermatozoa from frozen-thawed buffalo semen was comp ared. The recovery rate of motile spermatozoa was 95% in filtration pr ocedure and 33% in swim-up procedure (P<0.005). In all experiments, no rmal acrosomes did not vary due to treatments (P>0.05). In conclusion, HEPES and CO:! had no significant effect on swim-up of buffalo sperma tozoa. The pH 8.5 of TALP improved the recovery rate of motile spermat ozoa in swim-up procedure. The ion-exchange filtration was found super ior to swim-up procedure in harvesting maximum number of motile sperma tozoa from frozen-thawed buffalo semen (95 vs 33%; P<0.001). (C) 1998 by Elsevier Science Inc.