Antioxidant effects of butylated hydroxytoluene on evaluated parameters of cryopreserved buffalo spermatozoa

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     The study was carried out to evaluate the potential impact of butylated hydroxytoluene (BHT) on the frozen–thawed semen quality of buffalo. Ejaculated semen was extended in a Tris–citrate egg yolk and bioexcell extenders containing various concentrations of BHT (0.5, 1.0, 2.0 and 3.0 mM). After cooling at 5 ºC, semen samples were frozen in liquid nitrogen vapor and plunged into liquid nitrogen for storage. Straws from each treatment were thawed to assess the semen quality in terms of sperm motility, viability, plasma membrane integrity and acrosomal integrity at 0 and 6 h after thawing. Post-thawed sperm motility was determined using a phase-contrast microscope. Viability, plasma membrane integrity and acrosomal integrity were evaluated by the supravital staining, hypo-osmotic swelling test and normal acrosomal reaction, respectively.  Adding BHT in two extenders conferred better cryopreserving ability to spermatozoa compared to control groups. The highest (P < 0.05) value was achieved by addition of 2.0 mM BHT upon thawing and 1.0 mM at 6 h after thawing. However, higher concentration of BHT (3.0 mM) reduced the some evaluated parameters compared to other concentration used. 

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