Effect of Ashwagandha (Withania somnifera) supplementation on oxidative stress and freezability of breeding bull semen

Withania somnifera (ashwagandha), a well-known Ayurvedic adaptogen, is recognized for its potent antioxidant, cytoprotective, and anticancer properties. Although its efficacy in improving semen quality under oxidative stress is well documented in humans, its potential application in bull semen cryopreservation, where oxidative damage is a key determinant of reduced sperm survival and function, remains largely unexplored. The present study provided the first experimental evidence on the effect of ashwagandha supplementation in bull semen extenders. A total of 18 bull ejaculates (n = 18; six per bull) were cryopreserved in tris-based extender and divided into four groups: control (no supplementation) and three treatment groups supplemented with 1%, 2.5%, and 3.5% (v/v) aqueous ashwagandha extract. Post-thaw assessments included progressive motility, viability, acrosome integrity, plasma membrane integrity, and oxidative stress markers [reactive oxygen species (ROS) and malondialdehyde (MDA)]. Supplementation at 2.5–3.5% significantly (p < 0.05) improved motility, viability, acrosome- and plasma membrane integrity, while reducing ROS and MDA levels. Although DNA fragmentation did not differ statistically, numerical improvements were observed. The results demonstrated that ashwagandha supplementation enhanced post-thaw semen quality and mitigates cryo-induced oxidative stress in breeding bulls. Overall, the findings established Withania somnifera as a promising natural additive for bull semen extenders, with practical potential to improve the freezability and functional resilience of bull spermatozoa.