D uring the semen cryopreservation process, the characteristics of mammalian spermatozoa decreased because of the subject of spermatozoa to different stresses like the cold shock which may lead to damage of the mitochondria, plasma membrane, and acrosome membrane of sperm cells (Smith et al., 2011;Perteghella et al., 2017;Zhang et al., 2019). In bull semen, the abundance of unsaturated fatty acids in the sperm plasma membrane causes its exposure to oxidation, leading to damage of the sperm membrane, DNA fragmentation, and reduced enzyme activity (Kadirvel et al., 2009; El-Regalaty, 2017), consequently reduces motility and livability of spermatozoa (Asadpour et al., 2012; Gualtieri et al., 2014). Comparing to cattle, the buffalo semen is highly affected by cryopreservation damage during freeze-thawing processes in terms of reducing its fertilization ability (Andrabi, 2009), and decreasing the conception rate of buffaloes (Guthrie and Welch, 2012).The antioxidants are compounds that could decrease the harmful effects of oxidative stress on spermatozoa during the sperm preservation process. Antioxidants have a scavenging role for reactive oxygen species (ROS) that produced from lipid peroxidation (LPO) by improving the viability and fertilizability potential of buffalo spermatozoa (Lone et al., 2018).