Cerium oxide nanoparticles improve the post-thaw quality and in-vivo fertility of Beetal buck spermatozoa.

The motility, health quality, and membrane disorders of spermatozoa are adversely affected during the process of semen cryopreservation due to the over-production of reactive oxygen species (ROS). Cerium oxide nanoparticles (CeONPs) possess properties to scavenge ROS either by mimicking specific antioxidants or by enhancing the activities of antioxidant enzymes. Therefore, we aimed at evaluating the effects of adding the CeONPs in the TRIS-citrate-yolk extender on in-vitro antioxidant enzyme activities, spermatozoa quality attributes, and in-vivo fertility of post-thaw Beetal buck spermatozoa. The CeONPs were prepared and characterized (UV-spectrophotometry, FTIR, and XRD). Semen samples, collected from bucks (n = 5), were distributed into five aliquots and diluted in an extender containing increasing concentrations of nanoparticles (0 μg/ml, called the control group, 25 μg/mL, 50 μg/mL, 75 μg/mL, and 100 μg/mL). At post-thaw, spermatozoa were evaluated for the above-mentioned attributes and the pregnancy rate by inseminating Beetal does (n = 252). Results demonstrated that CeONPs mitigated the detrimental effects of cryopreservation as ROS production and lipid peroxidation were lower (P < 0.001) in the 25, 50, and 75 μg/mL CeONPs-added groups compared to the control and 100 μg/ml CeONPs-added group. The addition of 25 μg/mL CeONPs improved (P < 0.001) the activities of superoxide dismutase, catalase, and peroxidase and the concentration of reduced glutathione (P < 0.001) compared to the other groups. In terms of sperm kinematics and velocity parameters, the groups added with the 25 and 50 μg/mL CeONPs exhibited higher total motility (P < 0.001), sperm progressive motility (P = 0.003), and rapid velocity (P < 0.001). The group added with the 50 μg/mL CeONPs had the highest (P = 0.04) average path velocity. The groups added with the 25 and 50 μg/mL CeONPs also exhibited higher plasma membrane integrity (P = 0.003), acrosomal integrity, and viability (P < 0.001) compared to the control group. The DNA integrity was also higher (P < 0.001) in all the CeONPs-added groups. The pregnancy rate was higher (P = 0.003) in the 25 (51.92 %) and 50 μg/mL CeONPs (58.33 %) groups compared to the other groups. Conclusively, our findings suggest that the inclusion of cerium oxide nanoparticles in the TRIS-citrate-yolk freezing extender can reduce the occurrence of cryopreservation-induced damages to Beetal's buck spermatozoa and ultimately enhance the pregnancy rate in does.