ABSTRACT
In the present study, two experiments were conducted. The first experiment aimed to study the influence of buffalo bull semen filtration throughout different grades of sephadex columns [G-25, G-50, G-75, G-100 and G-200] on semen quality. Semen samples were collected from four buffalo bulls, diluted 1:20 with Tris buffer, loaded in different grades of sephadex columns and kept for 4-5 minutes at 37ooC. Filtered semen samples showed significantly [p<0.0001] higher individual motility [68.75 84.38%] in different grades of sephadex when compared to control samples [63.13%]. Percentages of live spermatozoa were found to be higher for filtered samples [71.38 -88.63%] than that of control samples [65.94%]. Percentages of major and minor abnormal spermatozoa were found to be decreased [2.37 4.5% and 1.56 5.12%, respectively] significantly [p< 0.0001] than that of control samples [5.56 and 6.31%, respectively]. Percentages of spermatozoa with acrosomal defects were found to be significantly [p<0.0001] lowered [1.44 3.88%] significantly [p<0.0001] in comparison with control samples [4.63%]. The loss in sperm number [0.67 to 0.99 vs 1.25 X 109 /ml] could be adequately compensated by obtaining higher values of all other important semen quality traits. The use of G-75 sephadex grade is recommended for wider use because it gives a more balanced picture of semen quality compared with other grades studied. The second experiment aimed to study the efficacy of sephadex G-75 filtration of buffalo spermatozoa for cryopreservation. Semen samples were collected from the same bulls used in the first experiment. Split samples were diluted 1:20 with Tris buffer, filtered through sephadex G-75 columns and centrifuged at 1500 rpm for 5 minutes. The supernatant was discarded and the sediments were re-diluted and frozen in Obtidyl diluent [Tris based diluent]. Filtration was found to improve semen quality after dilution, freezing and thawing. Post-dilution, filtrated sperm motility was significantly [p<0.0001] improved [86.25%] when compared with control samples [67.91%]. Post-thaw sperm motility of filtered samples was significantly [p<0.001] higher [61.25%] than that of control samples [45.83%]. Filtration was significantly enhanced sperm viability after thawing [p<0.0001], where viability indices were averaged 151.71 vs 90.83 for control samples. Percentages of spermatozoa with acrosomal defects were significantly [p<0.0001] reduced for filtered samples after dilution [1.92 vs 5.83%] and after freezing and thawing [7.75 vs 14.66%]. Bull individualities were found prominent for some parameters; post-thaw sperm motility and viability indices [p<0.05]. From the present study, filtration of semen samples through sephadex G-75 column could be recommended to improve pre- and post-thaw sperm quality of buffalo bull semen samples