The present study was undertaken to assess the effect of superoxide dismutase (SOD) on sperm motility, and viability; total sperm abnormality; acrosomal and plasma membrane integrity; DNA abnormality; antioxidant profiles such as catalase (CAT), reduced glutathione (GSH), and total antioxidant capacity (TAC); enzymatic profiles such as aspartate amino transaminase (AST), and alanine amino transaminase (ALT); and biochemical profiles such as malondialdehyde (MDA) production and cholesterol efflux. Total numbers of 50 ejaculates were collected twice a week from eight mithun bulls and semen was split into four equal aliquots, diluted with the TEYC extender. Group 1: semen without additives (control), and group 2 to group 4: semen was diluted with 50？U/mL, 100？U/mL, and 150？U/mL of SOD, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 1, 6, 12, 24, and 30？h of incubation. Inclusion of SOD into diluent resulted in significant ( ) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, SOD at 100？U/mL has significant improvement in quality of mithun semen than SOD at 50 or 150？U/mL stored in in-vitro for up to 30？h. It was concluded that the possible protective effects of SOD on sperm parameters are that it prevents MDA production and preserves the antioxidants and intracellular enzymes during preservation. 1. Introduction Mithun (Bos frontalis) is a semiwild free-range, rare bovine species present in the North-Eastern Hill (NEH) region of India. It is believed to have originated more than 8000 years ago from wild Indian gaur (Bos gaurus) [1]. The animal has an important place in the social, cultural, religious, and economic life of the tribal population particularly in the states of Arunachal Pradesh, Nagaland, Manipur, and Mizoram. Recent statistics indicates that the mithun population is decreasing gradually due to lack of suitable breeding bulls, increase in intensive inbreeding practices, declining land area for grazing, and lack of suitable breeding and feeding management in NEH region. Greater efforts are required from all quarters to preserve the mithun population to enhance the socioeconomic status of this region. Since mithuns are semiwild animals and not fully domesticated, natural breeding is practiced in this species with accompanied limitations like cost and disease transmission. Thus, use of AI for improvement of its pedigree is utmost essential. Cold storage
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