Effects of different concentrations of BHT on microscopic and oxidative parameters of Mahabadi goat semen following the freeze–thaw process

Abstract

Oxidative damage to sperm is one of the main causes for decline in motility and fertility of frozen–thawed sperm. Thus, it is crucial to use cryoprotectant agents in extender in order to prevent lethal intracellular ice crystal formation. The present study aims to evaluate the effects of different concentrations of the antioxidant butylated hyroxytoluene (BHT) on sperm parameters post-thaw. Semen was diluted into five equal aliquots of extender containing different concentrations of BHT (0, 0.5, 1, 2 and 4mM), aspirated into 0.25mL straws, and equilibrated at 5°C for 2h. After equilibration, straws were frozen in liquid nitrogen vapor and plunged into liquid nitrogen for storage. Sperm parameters, including motility and progressive motility, viability, membrane integrity, acrosome integrity and capacitation status, were assessed. Malondialdehiyde (MDA) and glutathione peroxidase (GSH-PX) activity were also evaluated after freezing-thawing. Results of this experiment show that addition of 1mM of BHT to the extender for freezing of goat semen can improve motility, progressive motility and viability (P<0.05) and reduce the MDA level (P<0.01). HOST (hypo-osmotic swelling test), acrosome integrity, capacitation status and GSH-PX were not affected by the concentrations of BHT (P>0.05). Therefore, we conclude that the optimum concentration of BHT for cryopreservation of goat semen is 1mM.