Optimal inclusion level of butylated hydroxytoluene in semen extender improves the quality of post-thawed canine sperm

Abstract

The study was conducted to evaluate the potential cryoprotective effect of butylated hydroxytoluene (BHT) through post-thaw evaluation of canine semen and its optimal inclusion level. Ejaculated canine semen was extended in TRIS-glucose egg yolk extender containing various concentrations of BHT (0.5, 1.0, 1.5, 2.0, and 2.5mM). Semen was frozen at -196°C using 200 × 106 spermatozoa per 0.5 ml straws and post-thaw evaluation was carried out in terms of sperm motility, viability, plasma membrane integrity, and acrosomal integrity through phase-contrast microscope, supravital staining, hypo-osmotic swelling test, and normal acrosomal ridge, respectively. BHT was found to improve (P > 0.005) all post-thawed semen quality parameters at an inclusion level of 1.0mM in the extended semen. However, higher concentrations than this were found to have detrimental effects.