Abstract
The aim of the present study was to compare the efficiency of various conventional single and dual staining protocols for assessment of spermatozoa attributes. The efficiency of single stains such as Eosin-Nigrosin (EN) and Trypan blue (TB) for viability and Giemsa (G) for acrosome integrity vis-a-vis dual stains such as Trypan blue-Giemsa (TB-G) and Eosin Nigrosin-Giemsa (EN-G) were compared. Ejaculates (12) from 3 buffalo bulls were used for evaluation of livability and acrosome integrity using said stains. The smears stained with EN-G were difficult to visualize distinctly and provided obscure results. A nonsignificant difference was found between EN, TB and TB-G for spermatozoa viability. However, at recommended concentration, TB yielded faint images, thus making the observation unnecessarily tedious. However, staining with TB-G was found advantageous for simultaneous determination of viability and acrosome status. Moreover, the results between G and TB-G deferred nonsignificantly indicating similar effectiveness of both the stains. Overall, TB-G can be a reliable, quick and convenient method for determining viability and acrosome integrity simultaneously for routine semen evaluation.
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