author(s)
Visakh Viswam, K Loganathasamy, VS Gomathy and D Reena

abstract
The present experiment was conducted to study the ameliorative effects of exogenous supplementation of osteopontin (ONP) on sperm morphology of frozen thawed buffalo semen treated with sodium nitroprusside (SNP), a nitric oxide (NO) donor. Buffalo semen straws from 8 ejaculates in 6 bulls were procured from Central Frozen Semen Production and Training Institute, Hessarghatta, Banglore and stored at Semen Bank, Madras Veterinary College, Chennai. The frozen straws were thawed and seminal plasma and semen extender were removed from spermatozoa by centrifugation. Spermatozoa were suspended in 1mL capacitation medium (control), addition of 100µg/mL of OPN (treatment I), 100µM/mL of SNP (treatment II) and 100µg/mL of OPN + 100µM/mL of SNP (treatment III). Semen with capacitation medium without any treatment served as control. The contents were incubated at 37ºC for 4 h and the sperm morphology was determined by Rose Bengal stain technique.No significant difference was observed in morphologically normal spermatozoa between control (81.75% ± 0.16) and treatment I (83.85% ± 0.20). Among treatment groups, significantly (P<0.05) more morphologically normal spermatozoa were observed in treatment I as compared to treatment II (57.05% ± 0.07) and III (68.50% ± 0.09). On comparison with control, significantly (P<0.05) lesser number of morphologically normal spermatozoa were found in treatment II and III. Morphologically normal spermatozoa in treatment III were significantly (P<0.05) higher than treatment II. The results of this study revealed that SNP supplementation alone have detrimental effects on the sperm morphology and co-incubation with OPN partially ameliorated the toxic effects of SNP on the sperm morphology of frozen thawed buffalo semen.