Evaluation of Microscopic, Flow Cytometric, and Oxidative Parameters of the Frozen–Thawed Bull Sperm in a Freezing Extender Containing Myo-Inositol

Introduction: This research was conducted to assess the effect of myo-inositol (MYO) in the freezing extender on the semen quality and oxidative stress parameters of frozen-thawed bull sperm. Materials and Methods: Semen samples were obtained from four bulls (n = 24, six ejaculates per bull), twice a week, and diluted into four equal aliquots in freezing extenders containing different concentrations of MYO (0, 2, 3, and 4 mg/mL). After a freezing/thawing process, velocity parameters, plasma membrane integrity, apoptosis status, malondialdehyde level, and oxidative stress parameters were assessed. Results: Supplementation of freezing extender with 3 mg/mL MYO resulted in higher rapid motility (62.22% ± 2.63%), progressive motility (77.45% ± 2.65%), viability (78% ± 0.91%), plasma membrane integrity (86 ± 0.85), catalase (20.03 ± 0.39 U/mL) activity, and lower significance of lipid peroxidation (3.60 ± 0.15 nmol/dL) than those of the control group (p < 0.05). A significantly lower percentage of normal morphology and intact acrosomes were observed for frozen-thawed semen in the extender supplemented with 4 mg/mL MYO than those of the control group (p < 0.05). Freezing of the sperm in the extender containing 3 mg/mL of MYO leads to a higher percentage of live cells (38.3 ± 2.76). Beat-cross-frequency, amplitude of lateral head displacement, linearity, total antioxidant capacity, total peroxidase activity, early apoptotic status, and superoxide dismutase activities were not affected by MYO levels in the extenders (p > 0.05). Conclusion: The findings of this study suggest that the supplementation of the freezing extender with 3 mg/mL MYO resulted in a higher quality of frozen-thawed bull sperm.