Comparison of normal and dual staining techniques for evaluating viability and acrosome status in frozen thawed buck spermatozoa

A study was conducted to assess viability and acrosomal integrity individually as well as simultaneously in the same slide using 12 frozen semen samples from adult Jamunapari bucks maintained at this Institute. The mean per cent live and acrosome intact (A), live and acrosome non-intact (B), dead and acrosome intact (C) and dead and acrosome non-intact (D) spermatozoa in dual staining technique (Eosin-Nigrosine and Giemsa staining) were 49.13±0.59, 1.16±0.08, 1.77±0.04 and 47.94±0.50, respectively. The mean per cent live sperm count, dead sperm count, acrosome intact spermatozoa and acrosome non-intact spermatozoa in dual staining technique were 50.29±0.56 (A+B), 49.71±0.56 (C+D), 50.90±0.70 (A+C) and 49.10±0.70 (B+D), respectively. The corresponding figures in normal staining procedure were 49.27±0.63, 50.73±0.63, 49.99±0.55 and 50.01±0.55, respectively. The results obtained in the study did not differ significantly (P<0.05) between dual and normal staining technique. Hence the dual staining technique can be easily and routinely adopted in assessing the viability and acrosome integrity of buck spermatozoa simultaneously in the laboratory to economize chemicals as well as time.