In this study investigated the impact on Epimedium (Epimedium sp.) added to the sperm diluent during cryopreservation. For this purpose, semen was collected from Zavot bulls (n = 3) using the artificial vagina method and pooled. The pooled semen was diluted with a TRIS-based diluent and divided into four groups. The groups were divided into control and three Epimedium groups (10, 20, 30 µL; E1, E2, E3) and frozen in nitrogen solution vapor (approximately − 120 °C) and sretained in nitrogen solution for evaluation. Following a one-week storage period, the spermatozoa were thawed at 37 °C for 30 s, and sperm motility and kinematics (CASA), plasma membrane integrity (HOST), acrosome integrity (A), mitochondrial membrane potential (MMP), malondialdehyde (MDA) ratio, and glutathione peroxidase (GSH) activities were analysed. It was determined that adding 30 µL of Epimedium to the diluent increased sperm motility and kinematics compared to the control (p < 0.05). At the end of the study, Epimedium groups showed more favourable results than the control group in all assessments. Consequently, it was concluded that Epimedium added to the sperm diluent improved spermatological characteristics.
