Eco-friendly Synthesized Zinc Oxide Nanoparticles Improved Frozen-thawed Semen Quality and Antioxidant Capacity of Rams

Zinc oxide nanoparticles (ZnO-NPs) have antimicrobial, antioxidants, and anticancer properties. This study aimed to improve the post-thawing semen characteristics of rams using three concentrations of ZnO-NPs. ZnO-NPs in 0.0, 0.5, 1.0, 1.5 mg/ml tris-based diluents were used to extend semen collected from five Awassi rams twice weekly for two months. Post-thaw semen evaluation and measurement of Malondialdehyde (MDA), nitric oxide (NO), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), total cholesterol (CHO), low density lipoproteins (LDL), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), zinc, and copper (Cu). With increasing ZnO-NPs concentrations, the post-thaw sperm total motility (STM %), progressive motility (SPM) %, viability (SV%), functional membrane integrity (FMI %), acrosome integrity (AI %), NO, LDH, ALP, Cu, and zinc ascended linearly (P≤0.0001) but the abnormal sperm (AS%) and SOD decreased (P≤0.0001). The addition of 1.0 mg/ml ZnO-NPs increased (P≤0.0001) GPx but decreased (P≤0.0001) CHO and LDL. All concentrations of ZnO-NPs preserved (P≤0.0001) high AA compared to 0.0mg/ml and did not influence MDA. The addition of 0.5 mg/ml ZnO-NPs reduced (P≤0.059) Cu concentrations. All semen characteristics had strong positive correlations with zinc, LDH, and ALP and negative correlations with SOD. AS % had negative correlations with zinc, ALP, and LDH and positive correlation with SOD. All the three added concentrations of ZnO-NPs improved the post-thaw semen motility, viability, fertilizing capacity, and antioxidant capacity but 1.5mg/ml semen diluents proved the best concentration for semen preservation.