Effect of butylated hydroxytoluene on bull spermatozoa frozen in egg yolk-citrate extender

Effect of butylated hydroxytoluene (BHT) on the quality of frozen-thawed Holstein bull sperm in egg yolk-citrate extender was evaluated. High quality semen samples were diluted in egg yolk-citrate extenders containing 0, 0.5, 1, 2 and 4 mM BHT and subsequently frozen in liquid nitrogen. Pre-freeze and post-thaw progressive motility, and live/dead ratio and acrosomal integrity of 200 sperm per slide, stained with Eosin-Nigrosin and Giemsa, were evaluated at 0, 2 and 4 h after thawing. There was a significant decrease in forward motility, livability and acrosomal integrity up to 4 h after thawing the frozen sperm. Upon thawing, sperm progressive motility at 1 mM BHT was significantly (P<0.001) higher (11%) than other groups, but percentages of live sperm and live sperm with intact acrosomes were higher at 0.5 mM BHT. BHT at 4 mM BHT caused a significant decrease in motility, livability and acrosomal integrity during preparatory stages of freezing sperm. It is concluded that 0.5-1.0 mM BHT can be beneficial for freezing Holstein bull spermatozoa in egg yolk-citrate diluent, when inseminated immediately after thawing.