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In this study, the role of melatonin has been explored for the prevention of Cerastes cerastes gasperettii venom-mediated toxicity and oxidative injury in male mice. The horned viper, C. c. gasperettii, is the most common snake in the subtropical deserts, including Saudi Arabia; its bite can be fatal. Therefore, we investigated the potential role of melatonin (10 mg/kg body wt) against the LD50 of C. c. gasperettii venom (0.978 mg/kg body wt) after 1, 2 and 6 h. The in vivo oxidative injury induced by the venom was clearly evident by the increased oxidative stress markers (lipid peroxidation and nitrite/nitrate) and decreased antioxidant molecules/enzymes (glutathione, superoxide dismutase and catalase), along with the histopathological alternations in liver, kidney and lung. In addition, mice envenomation disturbed the normal serum levels of liver and kidney function parameters. These biochemical changes and pathological alternations prior to C. c. gasperettii venom injection were effectively counteracted by melatonin. In conclusion, these results clearly indicate that melatonin attenuates oxidative stress, pathogenesis and dysfunction of liver, kidney and lung of mice in C. c. gasperettii venom-induced toxicity and oxidative injury via its antioxidant property. Therefore, it may be useful for the treatment for C. c. gasperettii bites.