The functional roles of plant extracts have been investigated for the treatment of various diseases including subfertility. Recent studies have highlighted the benefits of ashwagandha extract (AE) in enhancing sperm production, boosting testosterone levels, and lowering reactive oxygen species (ROS) levels in mammals. The current study is to examine the effects of the addition of AE to liquid boar semen on sperm quality during storage and its potential application in assisted reproductive technology. A hot water extract of ashwagandha was prepared from the dried powder of ashwagandha roots. Boar spermatozoa were stored in Beltsville thawing solution (BTS) at 17℃ for 5 days, with various concentrations of AE (1–50 mg/mL). During storage, motility, viability, acrosomal integrity and ROS of boar spermatozoa were examined. The results have shown that sperm stored in BTS with varying quantities of AE ranging from 1–20 mg/mL exhibited higher motility compared to those without AE (control) or with 50 mg/mL AE for 5 days. Similarly, sperm viability was better maintained in sperm treated with 1–20 mg/mL AE. Moreover, sperm stored in BTS with AE led to significantly higher acrosomal integrity and chromatin stability rates than sperm stored without AE. Notably, intracellular ROS levels significantly decreased in sperm stored in BTS with AE. Particularly, spermatozoa stored at 10 mg/mL AE exhibited an effective reduction in ROS during storage. These findings suggest the potential role of AE as an additive during sperm storage maintains sperm quality and can be used during subfertility treatment in both animals and humans.

INTRODUCTION
MATERIALS AND METHODS
    Preparation of ashwagandha extract
    Preparation of liquid boar semen
    Sperm motility evaluation
    Sperm viability
    Acrosome integrity
    Sperm chromatin stability
    Measurement of reactive oxygen species
    Statistical analysis
RESULTS
DISCUSSION
References

• Malavi Arachchi Gamage Nayodya Dananjanee Malavi Arachchi Gamage(Department of Agricultural Education, College of Education, Sunchon National University, Suncheon 57922, Korea)
• Wijesooriya Mudhiyanselage Nadeema Dissanayake(Department of Agricultural Education, College of Education, Sunchon National University, Suncheon 57922, Korea)
• Young-Joo Yi(Department of Agricultural Education, College of Education, Sunchon National University, Suncheon 57922, Korea) Corresponding author