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Oxidative stress associated with spermatozoa cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Lycopene, the most powerful singlet oxygen quencher of all carotenoids, may be a possible option for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of lycopene on selected oxidative stress parameters in cryopreserved bovine sperm. Ten bovine ejaculates were splitted into two aliquots and diluted with a commercial semen extender containing lycopene (1.5 mM/L) or no supplement (control), cooled to 4°C, frozen and kept in liquid nitrogen. Frozen straws were thawn in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, reactive oxygen species (ROS) generation was quantified using luminometry and LPO was assessed via the TBARS assay. Lycopene supplementation significantly (P<0.001) increased the spermatozoa motility and provided a significantly (P<0.001) higher protection against ROS overgeneration caused by semen freezing and thawing. It was determined that the presence of lycopene resulted in a significantly (P<0.001) lower LPO of the spermatozoa membranes. In conclusion, lycopene supplementation may be recommended to facilitate the improvement of semen preservation in bovine breeding industry.