Effect of l-Arginine and spermine-NONOate on motility, viability, membrane integrity and lipid peroxidation of Murrah buffalo (Bubalus bubalis) spermatozoa

In the present study freshly ejaculated Murrah buffalo semen samples of mass motility of +3 or above on a scale of 0-5 were collected and cryopreserved with l-Arginine (1 mM), spermine-NONOate (10 μM) and no additive. Post-thaw sperm motility, viability, membrane integrity and lipid peroxidation were assessed. Cryopreserved samples without any additive showed significantly (P<0.05) lower motility (32.33±1.45%) as compared to the cryopreserved samples treated with l-Arginine (38.33±1.66%) and spermine-NONOate (43.33±1.45%). Sperm viability, membrane integrity and mitochondrial activity were evaluated by eosin-Y staining, hypo osmotic swelling test and MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide] assay respectively. Lipid peroxidation was significantly (P<0.05) increased after freeze-thawing in untreated samples and decreased significantly (P<0.05) in spermine-NONOate (3.11±0.08 nmoles/108 cells) and l-Arginine (3.50±0.08 nmoles/108 cells) treated samples. The results therefore, clearly indicate that l-Arginine and spermine-NONOate play important role in protecting the post-thaw sperm motility, viability, membrane integrity and lipid peroxidation status in the cryopreserved Murrah buffalo spermatozoa.