Antioxidative Capacity of L-ascorbic Acid and Quercetin on Sperm Functional Integrity during Cryopreservation of Gir Bull Semen under Semi-arid Conditions of Rajasthan

Aim: The main objective of the study was to evaluate the antioxidative efficacy of L-ascorbic acid, quercetin and their combination (L-ascorbic acid + quercetin) on standard semen quality parameters including Sperm motility (%), Sperm viability (%), Abnormal Spermatozoa and acrosomal integrity in cryopreserved Gir bull semen. Methodology: A total number of 24 ejaculates were collected and evaluated. Ejaculate was processed separately and divided into four equal aliquots designated as Control (no antioxidant), T-1 (L-ascorbic acid @ 2.5 mM), T-2 (quercetin @ 100 µM), and T-3 (L-ascorbic acid @ 2.5 mM + quercetin @ 100 µM). Samples were cooled to 5 °C, packed in 0.25 ml French straws, frozen, and stored in liquid nitrogen (−196 °C). Frozen straws were thawed individually at 37 °C for 30 second before evaluation. Results: Significant differences (P < 0.05) were observed among the treatment groups for all evaluated semen quality parameters, including Progressive motility (%), Live spermatozoa (%), Intact acrosome (%) and Abnormal spermatozoa (%) at all stages of cryopreservation including post-dilution (PD), post-equilibration (PE), and post-thaw (PT) stages. Among the treatments, the combination group (T-3) exhibited significantly superior results compared to the individual antioxidant treatments (T-1 and T-2), whereas T-2 showed significantly better performance than T-1 for most of the evaluated parameters. Conclusion: The present study concluded that the combination of L-ascorbic acid and quercetin exerts the most pronounced protective effect against oxidative stress and cryoinjury, compared to their individual supplementation, thereby improving semen quality during cryopreservation in Gir bulls.