Authors : Abdolreza Rastegarnia, Neda Shetabi, Tohid Rezaei Topraggaleah and Yosef Nasiri
Abstract: The study was designed to investigate the effect of exogenous glutathione supplementation in soybean based extender Bioxcell® extender on post thaw semen quality of buffalo (Bubalus bubalis). Split pooled ejaculates (n = 6), possessing >70% visual sperm motility were extended at 37°C with different levels of glutathione (0.0, 0.5, 1 and 2 mM) in Bioxcell® extender. Semen was cooled to 4°C in 2 h, equilibrated at 4°C for 4 h, filled in 0.5 mL straws and frozen in a programmable cell freezer before plunging into liquid nitrogen. Thawing of frozen semen was performed after 72 h at 37°C for 30 sec. Sperm motion characteristics, viability, plasma membrane integrity, acrosome morphology of each semen sample immediately after thawing and incubation for 2 h were assessed by using Computer assisted semen analyzer (SCA), eosin-nigrosin staining, Hypo Osmotic Swelling (HOS) assay and phase contrast microscope, respectively. Results revealed that the addition 0.5 and 1.0 Mm of glutathione in Bioxcell® extender did not present any significant effect on overall and progressive motility as well as sperm motility characteristics (VAP, VSL, VCL, LIN and ALH), compared to the control groups (p>0.05). Immediately after thawing the proportion of post thaw sperm viability, plasma membrane integrity and normal apical ridge remained similar in all groups. However, glutathione supplementation of the extender with 2.0 mM concentration decreased sperm motility, viability at 0 and 2 h after thawing in a dose dependent manner compared to the control (p<0.05). The integrity of the sperm cells and percentage of spermatozoa with intact acrosome were not affected by the addition antioxidant (p>0.05). These results revealed that supplementation of the new commercial in soybean based extender Bioxcell® with glutathione did not improve sperm post thaw motility or acrosomal integrity.
Abdolreza Rastegarnia, Neda Shetabi, Tohid Rezaei Topraggaleah and Yosef Nasiri, 2012. Effects of Exogenous Glutathione Supplementation in Biocell® Extender on Quality of Cryopreserved Buffalo (Bubalus bubalis) Semen. Journal of Animal and Veterinary Advances, 11: 3437-3443.