Evaluation of Acrosomal Integrity and Viability in Bull Spermatozoa: Comparison of Cytochemical and Fluorescent Techniques

Abstract

This experiment was aimed to compare two well-known methods for assessment of acrosomal integrity and another two methods for assessment of viability in bull spermatozoa during cryopreservation. For this purpose, ejaculates were collected from four Hariana bulls using artificial vagina at biweekly interval. The semen samples which possesses more than 70% progressive motility and above 500 million/ml spermatozoa concentration was subsequently subjected to processing for liquid nitrogen (LN₂) vapour freezing. Semen samples were extended in Tris-egg yolk-glycerol extender and further subjected to two staining methods, cytochemical by Giemsa staining and fluorescent method by fluorescein isothiocyanate - pisum sativum agglutinin (FITC-PSA) at two stages i.e. pre freeze and post thaw. In the same way to assess the viability eosin-nigrosin staining is compared with fluorescent method i.e. Annexin-V/PI method. Results showed that fluorescent staining were more sensitive as compare to cytochemical method for spermatozoa of Hariana bulls.