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Aims: The present research was done to evaluate the antioxidant activity of aqueous extract of Withania somnifera (Ashwagandha) in equine. Herbal antioxidants have gained popularity due to their potency and safety, and oxidative stress is a condition that is more prevalent in horses performing physical activities. Methods: Twenty clinically healthy horses were selected and equally divided into treatment and control group consisting of 10 animals in each. An aqueous extract was prepared from dried root powder of Withania somnifera (Ashwagandha) and was continuously administered orally for 15 days to treatment group animals @ 100 mg/kg body weight. Blood samples were collected on 0 th, 8th, and 16th day from both the treatment and control group. The plasma was used for the determination of enzymatic oxidants (Catalase (CAT) and superoxide dismutase (SOD) and non-enzymatic antioxidants (reduced glutathione (GSH) and Vitamin C). Results: A non-significant increase in the values of Catalase, Superoxide Dismutase (SOD), Reduced Glutathione (GSH), and Vitamin C were observed after administration of aqueous extract of Withania somnifera. Conclusion: Administration of aqueous extract of Withania somnifera not significantly modulated the activity of enzymatic and non-enzymatic antioxidants.