Evaluation of three methods for use of bovine serum albumin in bull spermatozoa sexing

Abstract

This study was conducted to determine the possibility to vary the proportion of bull spermatozoa with Y chromosome by using bovine serum albumin (BSA) in three methods: Double centrifugation in BSA (M1); centrifugation and migration in BSA (M2); Double migration in BSA (M3). The sperm samples were stored for 20-24 hours at 5 °C and then frozen in liquid nitrogen. The variation in the proportion of spermatozoa with Y chromosome (f-body) was determined by staining with quinacrine dihydrochloride. No significant difference was observed in the percentage of f-body (48.4±5.9% for the control and from 45.8±5.5 to 47.2±4.9% for the treatments. The spermatozoa retrieval rate was 50.7% for M1, 36.2% for M2, and 21.4% for M3 (p<0.01). The effect of ASB on sperm characteristics was significant (p<0.01): the sperm vitality was reduced from 64.2% in the control to 42.5, 37.2 and 27.2% in M1, M2 and M3 respectively; motility was reduced from 83.1% in the control to 48.1, 31.9 and 23.1% in M1, M2 and M3 respectively; the percentage of sperm with functional membrane was reduced from 64.6% in the control to 42.7, 42.6 and 39.6% in M1, M2 and M3 respectively; and the percentage of live sperm with unreacted acrosome was 58.1% in the control and 35.1, 30.2 and 20.5% in M1, M2 and M3 respectively. Sperm characteristics of frozen semen were highly affected (p<0.01) making them unviable.