Supplementation of Mito TEMPO and acetovanillone in semen extender improves freezability of buffalo spermatozoa.

BACKGROUND: Oxidative stress is one of the leading factors responsible for poor post-thaw semen quality because of overproduction of reactive oxygen species (ROS) over neutralizing antioxidants present in semen. Mainly two ROS generation sites are present in spermatozoa, that is, mitochondria and plasma membrane. Therefore, the idea of targeting these specific sites for minimization of ROS production with the compounds having known mechanism of actions was built up as a core for this research. OBJECTIVE: Present study was done to investigate the effects of Mito TEMPO and acetovanillone individually and in combination on freezability of buffalo spermatozoa. MATERIALS AND METHODS: For the experiment, semen extender was supplemented with Mito TEMPO (50 µM), acetovanillone (50 µM), and a combination of Mito TEMPO + acetovanillone (50 µM+ 50 µM), designated as Group II, Group III, and Group IV, respectively. Control group without any supplementation was designated as Group I. A total of 24 ejaculates with individual progressive motility (IPM) of ≥70% were selected for the study. After final dilution, filling-sealing of straws, equilibration, and freezing were done as per the standard procedure. Semen samples were evaluated for IPM, plasma membrane integrity, lipid peroxidation, total antioxidant capacity (TAC), and cholesterol to phospholipids (C/P) ratio at both fresh and post-thaw stages. Evaluation of ROS, mitochondrial membrane potential (MMP), capacitation status (CTC assay), and in vitro fertility potential were conducted only on frozen-thawed samples. RESULTS: The addition of Mito TEMPO (50 µM) and acetovanillone (50 µM) individually and in combination significantly (p < 0.05) improved post-thaw semen quality in terms of IPM, plasma membrane integrity, TAC, cholesterol content, C/P ratio, MMP, Chlortetracycline (CTC)-Full (F) pattern, and zona binding ability of buffalo spermatozoa, while significantly (p < 0.05) reduced ROS production, lipid peroxidation, and capacitation like changes as compared to the control group. DISCUSSION: As Mito TEMPO acts as an SOD mimetic and also detoxifies ferrous iron at the mitochondria level, it aids in neutralization of excessive ROS production and minimizes oxidative stress-related damages that enhances the antioxidant potential of sperm mitochondria. Earlier studies also indicated improved post-thaw semen quality in 50 µM supplemented group. The improvement observed in acetovanillone (50 µM) group might be because of inhibition of Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase as this enzyme activation by various physical/chemical inducers during cryopreservation process leads to activation of CatSper channel resulting in calcium influx, premature capacitation, and acrosomal reaction like changes through activation of adenylate cyclase and cAMP/PKA-mediated tyrosine phosphorylation of sperm proteins. Acetovanillone also prevents NADPH oxidase-mediated inhibition of glutathione reductase activity, which has a vital role in protecting the structural and functional integrity of sperm plasma membrane. CONCLUSION: Results indicated beneficial effects of supplementation of Mito TEMPO and acetovanillone on sperm freezability and individual supplementation was as efficient as the combination group for sustaining post-thaw semen quality.

Body Weight and Scrotal-Testicular Biometry in Three Indigenous Breeds of Bucks in Arid and Semiarid Agroecologies, Ethiopia.

The body weight and testicular and epididymal parameters of Afar, Long-eared Somali (LES), and Woyto-Guji (WG) breeds of goat were investigated. A total of 405 randomly selected bucks of Afar (n = 135), Long-eared Somali (n = 135), and Woyto-Guji (n = 135) were included in this study. The overall mean scrotal circumference (SC), testicular volume (TV), testicular length (TL), testicular weight (TW), body weight (BW), epididymal weight (EW), body condition score, and testicular diameter (TD) measurements in all bucks were 20.8 ± 1.94 cm, 68.1 ± 6.18, 4.96 ± 0.79 cm, 70.0 ± 5.66 g, 22.1 ± 2.98 Kg, 9.09 ± 1.88 g, 2.55 ± 0.68, and 4.28 ± 0.45 cm, respectively. Significant (p < 0.05) breed differences in SC, TD, TL, TW, BW, EW, and TV were recorded. Long-eared Somali (LES) breed was heaviest and Afar breed was the lightest and Woyto-Guji (WG) had the average BW. In all breeds, the parameters were positively correlated. In Afar breed, the TW had a significant correlation with BW (r = 0.90) and SC (r = 0.65). In LES BW was highly correlated with TD (r = 0.96) and TL (r = 0.96). In WG, TW was significantly correlated with TD (r = 0.94), EW (r = 0.90), TL (r = 0.89), and BW (r = 0.82). In multiple regression analysis the linear combinations of BCS, SC, and BW significantly predicted TW, TL, TV, TD, and EW in all breeds. In conclusion, Long-eared Somali breed displayed greater BW and scrotal and testicular traits.