Lyophilized Coffee Grounds as a Natural Antioxidant to Preserve Post-Thaw Sperm Quality in Dorper and Creole Rams

Cryopreservation of ovine semen is associated with oxidative stress and structural sperm damage, which may compromise post-thaw sperm quality. Natural antioxidants derived from agro-industrial by-products have emerged as potential alternatives to reduce cryo-induced damage. The present study evaluated the effect of lyophilized coffee grounds extract (LCGE) supplementation on sperm quality during refrigeration and cryopreservation of Dorper and Creole ram semen. Semen samples from ten rams (five per breed) were allocated to three treatments based on LCGE concentration: 0 mg/mL (control), 1.0 mg/mL, and 1.5 mg/mL. Cryopreservation significantly affected all CASA-derived sperm parameters, with breed-dependent responses detected for several kinetic traits. LCGE supplementation had limited effects on CASA-derived sperm motility and kinetic parameters, with a significant dose effect detected only for total motility. In contrast, higher plasma membrane functionality, acrosomal integrity, DNA integrity, and chromatin condensation were observed in cryopreserved semen supplemented with LCGE, particularly at 1.5 mg/mL. LCGE-treated samples also exhibited a lower occurrence of moderate and high oxidative stress categories than cryopreserved control samples. Overall, the findings indicate that LCGE supplementation was associated with higher plasma membrane functionality, acrosomal integrity, DNA integrity, and chromatin condensation in cryopreserved semen. Further studies are required to determine the biological mechanisms involved and to evaluate the effects of LCGE supplementation on fertility under in vivo conditions.