BIOCHEMICAL CHARACTERIZATION OF FRESH AND FROZEN SEMEN OF SWAMP BUFFALO
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Date
2017-07
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College of Veterinary Science Assam Agricultural University Khanapara, Guwahati-781022
Abstract
Four Swamp buffalo bulls aged 5 to 8 years were used to study the biochemical characteristics of fresh and frozen semen and the effect of freezing on biochemical characteristics. The semen was subjected to protein profiling through SDS-PAGE in both fresh and frozen semen and their correlations with fresh and frozen semen characteristics were analyzed. A total of 60 ejaculates comprising 15 from each bull and a total of 40 ejaculates comprising 10 from each bull were used for evaluation and biochemical characterization in fresh and frozen semen respectively.
For studying the effect of freezing on biochemical constituents of semen the AST and ALT activities, total protein, total cholesterol and total lipid concentration were estimated in seminal plasma of fresh semen and extracellular fluid of frozen semen. The overall mean values of the AST and ALT activities and total protein, total cholesterol and total lipid concentration were 26.05±0.74 U/L, 49.14±1.72 U/L, 3.44±0.02 mg/ml, 23.12±0.65 mg/dl and 2.20±0.07 mg/ml respectively in seminal plasma of fresh semen. The corresponding values in extracellular fluid of frozen semen were 124.95±1.66 U/L, 87.80±2.29 U/L, 1.66±0.06 mg/ml, and 82.07±1.63 mg/dl and 32.70±0.74 mg/ml.
The mean value of AST and ALT activities and total cholesterol and total lipid concentration increased significantly (P˂ 0.01) and the total protein concentration was decreased significantly (P˂ 0.01) in extracellular fluid of frozen semen comparison with that in seminal plasma of fresh semen.
The proteins of Triton X-100-treated sperm membrane extract of fresh and frozen semen, seminal plasma of fresh semen and extracellular fluid of frozen semen were characterized by SDS-PAGE. Seven protein bands in seminal plasma and ten protein bands in extracellular fluid were identified. A total of eight and five protein bands were identified in sperm membrane extract of fresh and frozen semen respectively. The relative densities of sperm membrane proteins of different molecular weights as determined by Integrated Density Value were correlated with semen characteristics. The per cent initially motile sperm, live sperm percentage and intact acrosome of fresh and frozen semen were significantly correlated (P˂ 0.01) with sperm membrane proteins of different molecular weight in fresh and frozen semen. The study revealed that the 42, 55 and 104 kDa proteins of sperm membrane had significant correlation with semen characteristics and could be considered as potential marker in screening of swamp buffalo bulls.
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