The Effect of Adding Green Zinc Oxide Nanoparticles to Ram Semen Dilution Medium and Its Effects on Sperm Quality and Microbial Load of Frozen Semen

Introduction: Maintaining sperm quality and reducing contaminants will ensure the success of fertilization. The presence of bacterial contamination in the reproductive system of animals and their transfer to the semen causes a decrease in sperm quality and problems of disease transmission through inseminated. During the process of freezing and thawing, cold shock as well as environmental microbial pollution causes sperm quality to decrease. Zinc oxide nanoparticles, in addition to increasing the antioxidant capacity, also have antibacterial effects. The sperm membrane of ruminants are rich in unsaturated fatty acids and Zinc is as a cofactor of biological antioxidant biomolecules that suggested zinc increases protecting cell membrane and other inside organelles. Conventional chemical methods are expensive and require the use of chemical compounds/ solvents as reducing agents, which are toxic and as a risk factor for environment. Green chemistry reduces the risk of pollution at the source level, and instead of producing waste, it can use waste materials as a source of producing nanoparticles. This technology focuses on selecting reactions (for example biological molecules such as polyphenols, flavonoids, carbonyl and protein compounds) that are compatible with the environment.Materials and Methods: In this research, green zinc oxide (gZnO) nanoparticles are produced from saffron petals. Sperms were collected from 6 Afshari rams and after checking and confirming the quality, the sperms were mixed together in a 4x4 factorial design with 4 levels of antibiotics (0, 50, 75 and 100) recommended percentage and 4 levels of gZnO (0, 7.5, 10 and 12.5) μg/ml were added to the each diluent and entered to the freezing process. In order to check the quality of sperm after freezing CASA system were used. Membrane functionality was measured using osmotic method. The total microbial load was estimated using the colony count on blood-agar medium method.Results and Discussion: The results showed that 12.5 µg/ml level of gZnO nanoparticles becomes destructive and reduced quality of sperm, but the lower levels of gZnO significantly (P