ASSESSMENT OF THE EFFICACY O SEPHADEX FILTRATION OF BUFFALO SPERMATOZOA FOR CRYOPRESERVATION

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 45 minutes at 37eeC, 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) in comparison w'th control samples (4.63%), The loss in sperm number (0.67 to 0.99 vs 1.25 X 109 /mi) 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 1S1.71 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).