Sulforaphane inclusion in a freezing medium augments post-thaw motility, functional and biochemical features, and fertility potential of buffalo (Bubalus bubalis) spermatozoa

Abstract

Sulforaphane is a natural and highly effective antioxidant safeguarding the reproductive system, and alleviate oxidative stress. This study was designed in order to elaborate L-sulforaphane effect on semen quality, biochemical parameters, and fertility of buffalo (Bubalus bubalis) spermatozoa. Semen was collected from five buffalo bull with artificial vagina (42 °C) three times and evaluated for volume, consistency (color), motility, and sperm concentration. After critical examination, semen was diluted (50 × 106 spermatozoa per ml, 37 °C) in extenders with (2 μM, 5 μM, 10 μM, and, 20 μM) or without (control) sulforaphane, cooled (from 37 to 4 °C), equilibrated (4 °C), filled (straws, 4 °C), and cryopreserved (LN2, −196 °C). Data analysis exhibited that sulforaphane addition in extender augments total motility (%, 10 μM, and 20 μM than control), progressive motility (%), and rapid velocity (%, 20 μM than control), and velocity parameters (average path velocity, μm/s, straight line velocity, μm/s and curved linear velocity, μm/s, 20 μM than control, and 2 μM). Moreover, sulforaphane augments functional features (membrane functionality, mitochondrial potential, and acrosome integrity) of buffalo sperm (20 μM than control). Sulforaphane preserves biochemical features of seminal plasma of buffalo i.e., Calcium (μM), and total antioxidant capacity (μM/L), followed by reduction in lactate dehydrogenase (IU/L), reactive oxygen species (104 RLU/20 min/ 25 million), and lipid peroxidation (μM/ml) in 20 μM than control. Lastly, sulforaphane augments fertility rate of buffalo sperm at 20 μM than control, and 2 μM. Conclusively the existing study revealed that adding L-sulforaphane (20 μM) in a freezing medium augments motilities, kinematics, functional parameters, and fertility rate of buffalo spermatozoa. Correspondingly, sperm favorable biochemical features were also augmented with sulforaphane followed by reduction in oxidative stress parameters. Further studies are highly recommended to define the particular mechanism of action of sulforaphane in augmenting buffalo post-thawed semen quality, and in vitro fertility potential.