Adding kisspeptin-10 to the freezing solution for buffalo bull sperm helped protect the cells from damage during thawing. The best results were seen with 15‑20 µmol L⁻¹, which boosted antioxidant levels, reduced lipid oxidation, and improved sperm movement, membrane health, and DNA integrity.

Effects of kisspeptin-10 as antioxidant in cryodiluent were evaluated on post-thaw quality of buffalo spermatozoa. Qualified semen samples from five bulls were pooled, divided into five aliquots and extended in Tris-citric acid cryodiluent containing differential doses of kisspeptin-10 (5, 10, 15, and 20 μmol L^-1 and negative control. Extended sperm suspension was cooled to 4°C, packaged in 0.54 ml straws and cryopreserved. At post-thawing, catalase (unit mg^-1 ), peroxidase (unit mg^-1 ) and reduced glutathione (μmol L^-1 ) levels were highest (p < 0.05) with 20 μmol L^-1 of kisspeptin-10 as compared to negative control. Moreover, lipid peroxidation (nmol L^-1  min^-1  mg protein^-1 ) level was lowest (p < 0.05) with 20 μmol L^-1 of kisspeptin-10. Sperm progressive motility (%), rapid velocity (%) and kinematics were higher (p < 0.05) with 15 and 20 μmol L^-1 of kisspeptin-10 as compared to negative control. Supra-vital plasma membrane integrity (%), viable sperm with intact acrosome (%) and DNA integrity (%) were improved (p < 0.05) with all doses of kisspeptin-10 as compared to negative control. It was concluded that the addition of 15 and 20 μmol L^-1 kisspeptin-10 in cryodiluent ameliorated the overall frozen-thawed quality parameters of Nili-Ravi buffalo spermatozoa.