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Objective:To investigate changes in follicular fluid concentrations of reactive oxygen species (ROS) and total antioxidant capacity(TAC) and degree of oxidative damage to follicular cells, using protein carbonyl(PC) as marker of oxidative stress, were investigated during reproductive acyclicity in buffalo.Methods:Follicular fluid was aspirated from follicles grouped into three classes depending upon their diameter [small(5.0-7.0 mm), medium(7.1-10.0 mm), and large (>10.0 mm)].Progesterone and estradiol were estimated to determine functional status(P:E ratio) of the follicles.Results:Acyclic buffaloes had greater concentrations ofROS(P<0.001) andPC (P=0.0412) and lower concentrations ofTAC(P=0.0280) than cyclic buffaloes.An interesting novel finding was the complete absence of lowP:E functionally active follicles in acyclic buffaloes. Results indicated a pronounced follicular fluid oxidant-antioxidant imbalance and oxidative damage to follicular cells during acyclicity in buffalo.Conclusion:In conclusion, this study provided evidence about role of oxidative stress in pathogenesis of reproductive acyclicity.