Studies on Effect of Ascorbic Acid and Ashwagandha Supplementation on Post-thaw Sperm Quality and Oxidative Stress Status in Frieswal Bull Semen

Abstract

This experiment was designed to study the effect of ascorbic acid and ashwagandha on post thaw sperm quality and oxidative stress in Frieswal bull semen during cryopreservation. For this purpose, ejaculates were collected from three Frieswal bulls using artificial vagina at biweekly interval. The semen sample which possesses more than 70 % progressive motility and above 600 million/ ml spermatozoa concentration was subsequently subjected to processing for LN2 vapour freezing. Semen samples were extended in GEYT extender and split into 7 parts. One part was left as such (control) while the other 6 parts contain 2.5 mM (TG-I), 5.0 mM (TG-II) and 7.5 mM (TG-III) ascorbic acid and 20 μl/2ml (TG-IV), 50 μl/2ml (TG-V) and 70 μl/2ml(TG-VI) ashwagandha respectively. At fresh stage, semen was evaluated for volume, concentration, pH, per cent live spermatozoa, per cent progressive motility, per cent intact acrosome, per cent HOST positive spermatozoa. At post thaw stage semen was evaluated for per cent live spermatozoa, per cent progressive motility, per cent intact acrosome, per cent HOST positive spermatozoa, per cent DNA integrity, levels of LPO, levels of ROS. Supplementation of ascorbic acid and ashwagandha improves most of the parameters at post thaw and give a significant effect at the concentration of 5.0 mM ascorbic acid; 50μl/2 ml and 70 μl/2 ml ashwagandha in comparison to control and 2.5 mM and 7.5 mM concentration of ascorbic acid and 20 μl/2 ml ashwagandha respectively. Addition of ascorbic acid (5.0 mM) and ashwagandha (50μl/2 ml and 70 μl/2 ml) reduced oxidative stress during cryopreservation