Sodium alginate potentiates antioxidants, cryoprotection and antibacterial activities of egg yolk extender during semen cryopreservation in buffalo

Abstract

The study was conducted to determine effects of sodium alginate on sperm during cryopreservation. Each ejaculate (n = 20) of five buffalo bulls (3–5 years) were divided into six equal fractions and diluted using egg yolk based extender supplemented with different concentrations of sodium alginate and cryopreserved. Frozen-thawed semen samples were evaluated using the CASA, hypo-osmotic swelling test, cervical mucus penetration capacity test, and chlortetracycline fluorescence assay (CTC). Phosphorylation of tyrosine containing proteins and malondialdehyde concentration of sperm membrane were evaluated using immunoblotting and thiobarbituric acid reactive substance assay respectively. The semen extender’s anioxidative capacities were estimated by conducting 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays, metal chelating capacity by assessing ferrozine and antibacterial capacity using agar plate methods. Supplementation of sodium alginate in extender improved sperm longevity, plasma membrane integrity as well as capacity to transit through the cervical mucus. Supplementation of extender with sodium alginate minimises the phase transition of sperm membranes and phosphorylation of tyrosine containing proteins during cryopreservation. Malondialdehyde concentration of sperm was less in sodium alginate-treated sperm as compared with control samples. The results indicated that sodium alginate increased antioxidant capacity of semen extender. Supplementation with sodium alginate also improved the metal chelating capacity and antibacterial properties of the extender. In conclusion, supplementation of extender with sodium alginate enhances free radical scavenging, metal reduction and chelating capacities to protect sperm during cryopreservation.