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Superoxide Dismutase in Extender Improves the in vitro Quality and in vivo Fertility of Cryopreserved Water Buffalo (Bubalus bubalis) Spermatozoa. 2020
BACKGROUND Superoxide dismutase (SOD) as an antioxidant in semen extender may be used for the cryopreservation of buffalo spermatozoa and in vivo fertility. OBJECTIVE This study was aimed to evaluate the effects of SOD (SOD1, 100 IU/mL; SOD2, 200 IU/mL; SOD3, 300 IU/mL) and control (0.0) in Tris citric acid extender on in vitro quality and in vivo fertility of cryopreserved water buffalo bull spermatozoa. METHODS Semen collection was carried out on a weekly basis (four bulls, three replicates, and n = 24 ejaculates). The conventional freezing of semen loaded straws (0.5 mL) was undertaken by placing them horizontally on a steel rack inside a Styrofoam box for 10 min containing liquid nitrogen (LN2) vapours, and plunging into a liquid nitrogen tank (-196 °C) for storage, followed by thawing at 37 °C for 30 s and analysis by computer-assisted sperm analyzer (CASA) and other assays. RESULTS At post-dilution, the acrosome integrity (ACR-I, %) was significantly improved (P < 0.05) in extender supplemented with SOD3 as compared to other experimental groups. In addition, DNA integrity (DNA-I, %) was significantly higher (P < 0.05) in SOD1 and SOD3 compared to SOD2 and control. At post-thawing, the mean values of sperm progressive motility (PM, %), average path velocity (VAP, µm/s) and straight line velocity (VSL, µm/s) were significantly higher (P < 0.05) in extender supplemented with SOD3 compared to the control. At post-thawing, mean values of subjective motility (SM, %), plasma membrane integrity (PMI, %) and ACR-I were significantly higher (P < 0.05) in extender supplemented with SOD3 compared to the control. At post-thawing, sperm DNA-I was significantly higher (P < 0.05) in extender supplemented with all SOD doses compared to the control in a dose-dependent manner. The in vivo fertility rate (%) was significantly higher with SOD3 compared to the control (68.2 % vs. 49.5 %). CONCLUSIONS The supplementation of SOD3 (300 IU/mL) in Tris citric acid extender improves both in vitro quality and in vivo fertility of buffalo bull spermatozoa.
