Asphaltum Improves the Post-thaw Quality and Antioxidant Status of Nili Ravi Buffalo Bull Sperm

Abstract

Asphaltum, a mineral exudate from the mountains, is an ayurvedic medicine believed to be a panacea for male reproductive health issues. The objective of the study was to evaluate asphaltum in terms of phytochemical components, radical scavenging activity (RSA), in vitro dose tolerability, and cryosurvivability of buffalo sperm. Asphaltum was procured from an authentic source and confirmed for the presence of flavonoids, terpenoids, saponins, tannins, alkaloids, steroids, and glycosides. It showed good RSA as confirmed by the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. In vitro dose tolerability of buffalo sperm (n = 3, replicate = 4, ejaculates = 24) for asphaltum was assessed at 0.75%, 1.5%, 2.25%, 3.0%, 3.75%, 4.5%, 5.25%, and 6.0% (w/v). Buffalo sperm showed good tolerance up to 3% of asphaltum in terms of sperm progressive motility and plasma membrane integrity. Buffalo semen (n = 3, replicates = 4, ejaculates = 24) was cryopreserved in extender supplemented with 0.0%, 0.75%, 1.5%, 2.25%, and 3.0% (w/v) asphaltum and sperm quality was assessed at post-dilution, post-cooling, and post-thaw. After dilution motility, viability and livability; post-cooling motility and plasma membrane integrity; and post-thaw motility, plasma membrane integrity, viability, livability, DNA integrity, sperm RSA, sperm total lipids, sperm mitochondrial activity, and total antioxidant activity of semen were improved by 3%. In conclusion, asphaltum supplementation in an extender at 3% improves the post-thaw quality and antioxidant activity of buffalo semen.