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Castro et al. Journal of Animal Science and Biotechnology (2016) 7:17 DOI 10.1186/s40104-016-0076-xRESEARCHOpen AccessSperm cryodamage occurs after rapid freezing phase: flow cytometry approach and antioxidant enzymes activity at different get T0901317 stages of cryopreservationL. S. Castro1, T. R. S. Hamilton1, C. M. Mendes1,2, M. Nichi3, V. H. Barnabe3, J. A. Visintin1,2 and M. E. O. A. Assump o1*AbstractBackground: In order to improve the efficiency of bovine sperm cryopreservation process, it is important to understand how spermatozoa respond to differences in temperature as well as the ability to recover its own metabolism. The combination between flow cytometry approach and antioxidant enzymes activity allows a more sensible evaluation of sperm cell during cryopreservation. The aim of this study was to evaluate sperm attributes and antioxidant enzymes activity during different stages of cryopreservation process. Semen samples from Holstein bulls (n = 4) were separated in 3 treatments: fresh (37 ); cooled (5 ); and thawed. Evaluation occurred at 0 h and 2 h after incubation. Membrane integrity, mitochondrial membrane potential (MMP) and DNA damages were evaluated by flow cytometry; activities of antioxidant enzymes such as catalase, superoxide dismutase and gluthatione peroxidase were measured by spectrofotometry. Results: There was an increase in the percentage of sperm with DNA damage in the thawed group, compared to fresh and cooled, and for 2 hs of incubation when compared to 0 h. Considering MMP, there was.