This study was conducted to investigate the effects of different levels of α-tocopherol in cryopreservation media on the bull sperm quality after thawing. Semen samples were collected from six Holstein bulls using artificial vagina twice a week. Semen samples were pooled to eliminate individual differences and then divided into four equal parts for freezing with extenders containing different concentrations of α-tocopherol according to experimental groups as follows: 0 (control), 1.2 mM (E1), 2.4 mM (E2) and 4.8 mM (E4). Motion characteristics, viability, plasma membrane functionality, lipid peroxidation and H 2 O 2 status were determined after thawing. Results showed that malondialdehyde (MDA) concentration was significantly lower in E4 (6.1±0.6 nmol/mL) than E1 and E2 extenders (8.1±0.6 nmol/mL and 8±0.6 nmol/mL, respectively). Also, the lowest significant concentration of H 2 O 2 was observed in E4 (3.2±0.13 nmol/mL) compared to E1 (4 ±0.1 nmol/mL), E2 (5.3± 0.1 nmol/mL) and control (6.7±0.1 nmol/mL). Moreover, E2 and E4 produced the highest significant motility (74.2±1.6%, 75.9±1.6%), viability (78.2±1.8%, 76.1±1.8%) and membrane functionality (73±1.6%, 70.5±1.6%) compared to other groups. It can be concluded that α-tocopherol at the concentration of 4.8 mM can be an efficient antioxidant additive in Bioxcell extender for cryopreservation of bull semen.Additional key words: antioxidant; bioxcell; lipid peroxidation; sperm; H 2 O 2 .Correspondence should be addressed to Mohammad Chamani: m.chamani@srbiau.ac.ir Abbreviations used: LPO (lipid peroxidation); MDA (malondialdehyde); ROS (reactive oxygen species).Funding: The authors received no specific funding for this work.
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