Impact of Trolox Supplementation on the Cryopreservation of Honamli Buck Semen.

Abstract

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 in vitro 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 (p < 0.05). The control group had the lowest plasma membrane integrity (p < 0.05). The highest morphological integrity was observed in the T groups compared with the control group (p < 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.