The aim of this study was to investigate the effectiveness of hesperidin (HES) on testicular histopathological changes, biochemical changes, and semen characteristics in rats exposed to glyphosate (GLP). The control group was given a normal diet devoid of GLP and HES, the HES group was given 100 mg/kg/day HES with the normal diet, the GLP group was given GLP at the LD₅₀/10 dose of normal feed, which was 787.85 mg/kg/day, and the GLP + HES group was given normal feed containing 787.85 mg/kg/day LD₅₀/10 dose of GLP in addition to 100 mg/kg/day HES. GLP administration reduced sperm motility, sperm plasma membrane integrity, glutathione levels, and total antioxidant levels in the testicular tissues of rats. Moreover, it caused an increase in right testis and left epididymis weights, abnormal sperm counts, malondialdehyde levels, total oxidant status, and DNA damage. The HES treatment showed curative effects on these parameters. Furthermore, HES was effective in lessening the histopathological damage that was caused by GLP. The results showedthat HES protects spermatological parameters and DNA integrity, improves antioxidant defenses, and lowers the damage and lipid peroxidation caused by GLP in testicular tissue.

This study investigated the effects of supplementing Tris semen extender with protocatechuic acid (PCA) at doses of 25, 50, and 100 µg/mL on the quality of ram spermatozoa after cryopreservation. We evaluated motility, kinematic parameters, DNA integrity, redox status, plasma membrane–acrosome integrity (PMAI), viability, and high mitochondrial membrane potential (HMMP). PCA supplementation produced concentration-dependent and parameter-specific responses. The 25 µg/mL PCA group exhibited the most favorable functional profile, demonstrating improved progressive and total motility alongside reduced DNA damage. This protective effect was evidenced by lower tail length, tail DNA percentage, and tail moment in the comet assay. Conversely, treatment with 100 µg/mL PCA appeared to exert a notable effect on the redox balance, indicated by lower malondialdehyde, total oxidant status, and oxidative stress index levels, alongside an elevated total antioxidant status (p < 0.05). However, this higher concentration (100 µg/mL) did not provide parallel DNA protection and was associated with increased comet assay indices. PMAI, viability, and HMMP were not significantly affected by PCA supplementation (p > 0.05). These findings indicate that PCA exerts dose-dependent but parameter-specific effects on frozen–thawed ram spermatozoa. Low-dose PCA, particularly 25 µg/mL, appears more suitable for preserving motility and nuclear integrity, whereas 100 µg/mL mainly improves oxidative balance.