Abdul Azeez C. PMAHALINGESHWARA H. S.2020-07-172020-07-172018http://krishikosh.egranth.ac.in/handle/1/5810149298Binder of sperm protein (BSP), represents the major class of proteins in seminal plasma of ungulates. The proteins have been reported to play a major role in multiple aspects of fertilization including capacitation and sperm reservoir formation. In addition, they have been reviewed to influence the cryopreservability of bovine semen. The objective of this study is to characterise Fn2 proteins in Vechur bull semen, and to evaluate the effect of gelatin binding proteins (GBP) and non gelatin binding proteins (NGBP) of seminal plasma on freezability of Vechur bull semen. Semen collected from three Vechur bulls was centrifuged to separate seminal plasma. Gelatinagarose affinity chromatography was carried to isolate gelatin-binding proteins (GBP) and non gelatin-binding proteins (NGBP) from crude proteins. Isolated proteins were pooled separately dialysed against ammonium bicarbonate and lyophilized. SDS-PAGE of crude seminal plasma revealed presence of 15 bands. Out of which three were gelatin-binding proteins, in the molecular weight range of 14 to 16 kDa and one at 26 kDa. And the rest 12 bands were non gelatin binding proteins. For second phase of the study 18 ejaculates were collected from same three Vechur bulls, evaluated for fresh semen characters, pooled and washed with Tris buffer to remove seminal plasma and divided into four groups. Group I contain washed spermatozoa supplemented with NGBP (30 mg/ml) and GBP (30 mg/ml). Group II, Group III contain washed spermatozoa supplemented with NGBP (30 mg/ml), GBP (30 mg/ml) respectively, and Group IV contain washed spermatozoa without any seminal proteins. The semen in each aliquot incubated for 10 min. extended with TECY extender, equilibrated and frozen manually over liquid nitrogen vapour and stored in a cryocan. No significance difference was observed in sperm progressive motility and viability between the groups at pre-freeze and post thaw stage. Sperm abnormality per cent of was significantly (p<0.01) higher in group IV during pre-freeze and post thaw. Acrosome integrity, plasma membrane integrity and sperm membrane cholesterol were significantly (p<0.01) lower in group III at pre-freeze and post thaw stage. From the results it can be concluded that sperm supplimented with GBP at 30µg/ml reduced semen freezability, which can be ascribed to the deliterious effect of GBP.ennullThesis