Quality assessment of Y-enriched semen following Resiquimod protocol in Sahiwal bull

Abstract

The present study evaluated the effect of Resiquimod (R848)-based sperm sorting on the quality of Y-enriched semen in Sahiwal bulls. Semen was collected twice weekly from three healthy bulls, and 24 ejaculates meeting predefined quality criteria were processed. Samples were divided into untreated control (G-I) and R848-treated (G-II) groups. Sperm quality was assessed based on concentration, individual progressive motility (IPM), viability, morphological abnormalities, acrosome integrity, plasma membrane integrity using the hypo-osmotic swelling (HOS) test, and seminal plasma total antioxidant capacity (TAC). Following 30 min incubation with 0.6 µM R848, IPM did not differ significantly between groups (64.54 ± 0.79% vs. 62.08 ± 1.20%). Sorted semen showed significantly higher viability and lower sperm abnormalities, while acrosome integrity remained comparable to the control. HOS-positive spermatozoa and TAC were significantly reduced following sorting. Overall, R848-based sorting enriched functionally competent spermatozoa while maintaining key sperm attributes, suggesting its potential as a simple and cost-effective approach for Y-sperm enrichment.