Cryopreservation-induced oxidative stress significantly impairs sperm quality and fertility in buffalo semen. Quercetin is a natural antioxidant flavonoid known for its potent ability to scavenge reactive oxygen species and reduce oxidative stress. The present study investigated the oxidative stress biomarkers and DNA integrity in cryopreserved sperm following supplementation of various concentrations (0, 25, 50 and 100µM) of quercetin to a Tris-based extender. In this study, 32 semen ejaculates were collected from four sexually active buffalo bulls of Kundhi breed, twice weekly for four weeks. Semen ejaculates with ≥80% sperm motility were selected for processing. Ejaculates collected from four bulls on each collection day were pooled, diluted in Tris-based extender, cryopreserved and stored for 24h in liquid nitrogen. Following cryopreservation, the quercetin 100µM group showed the highest sperm viability (69.12±1.24%), plasma membrane integrity (62.25±1.08%), total motility (57.75±1.15%) and progressive motility (49.25±0.96%). Similarly, the activities of antioxidant enzymes-catalase (2.50±0.05U/mL), superoxide dismutase (50.625±1.02U/mL) and total glutathione (49.875±0.95µM) were significantly elevated, whereas malondialdehyde (MDA) levels (38.37±0.84nmol ml⁻¹) and DNA damage (8.625±0.42%) were significantly reduced, in the 100µM group compared with the control group (P<0.05). In vivo fertility trials demonstrated a higher conception rate in the quercetin-treated group (65%) compared with 40% in the control group (P<0.05). In conclusion, quercetin supplementation at 100µM to the Tris-based extender effectively alleviates oxidative stress during cryopreservation and enhances post-thaw sperm quality and improves fertility of semen of Kundhi buffalo bulls.

To Cite This Article: Malak KA, Korejo NA, Talpur HS and Kaka A, 2026. Effect of quercetin supplementation in tris-based extender on the post-thaw sperm quality, oxidative stress, DNA integrity and fertility of kundhi buffalo bulls. Pak Vet J, 46(6): 1618-1625. http://dx.doi.org/10.29261/pakvetj/2026.154