Impact of Trolox Supplementation on the Cryopreservation of Honamli Buck Semen

Cryopreservation of buck semen is essential in animal breeding but often damages sperm viability and integrity. The Honamli breed, a hardy Turkish goat, can benefit from improved freezing techniques using antioxidants such as Trolox (T). This study explores the effects of varying T concentrations on Honamli buck semen, assessing parameters such as motility, viability, and membrane integrity to enhance post-thaw quality. Findings support T's potential to improve semen extender formulations for preserving Honamli genetics.This study aims to freeze Honamli buck semen with T and to evaluate <i>in vitro</i> spermatological parameters. Three Honamli bucks, aged 2-3 years, were used in the study. Semen was collected from the bucks and mixed after removing seminal plasma. The mixed semen was diluted with a tris egg yolk extender containing three different concentrations of T (0.25 mM, 0.5 mM, and 1 mM) and control (0 mM). The diluted semen was equilibrated for 2 hours at +4 degrees and subjected to cryopreservation in liquid nitrogen vapor (-120°C for 12 minutes) and frozen. After thawing (37°C water bath for 30 seconds), the groups were evaluated at flow cytometric analysis for viability (SYBR/propidium iodide [PI]), plasma membrane acrosome integrity (FITC-PNA/PI), and mitochondrial membrane potential (JC-1), plasma membrane integrity (hypo-osmotic swelling test), microscopic evaluations for motility and morphological integrity (abnormal spermatozoa rate). The 0.5 T and 0.25 T groups showed significant improvements in motility compared with the control group (<i>p</i> < 0.05). The control group had the lowest plasma membrane integrity (<i>p</i> < 0.05). The highest morphological integrity was observed in the T groups compared with the control group (<i>p</i> < 0.05). In conclusion, supplementing T in buck semen extenders benefits spermatological parameters; particularly, 0.25 and 0.5 mM T could be used in Tris semen extenders during the cryopreservation process.