Stevia rebaudiana Bertoni has non-caloric sweetening properties and its use has been linked to therapeutic effects. However, Stevia sp is sold as a substitute for sugar commercially, not only includes steviosides but is also combined with other high-intensity artificial sweeteners, which questions its safety. Our objective was to evaluate the effect of natural and artificial Stevia rebaudiana Bertoni on fatigue and oxidative stress of skeletal muscle. Twenty-four male Wistar rats were divided into: (C) rats receiving water, (S); rats receiving 41.2 g/L sucrose solution (SRA); rats receiving solution with commercial sweetener (Svetia) 6 g/L; (SRN); rats receiving solution with the dried and powdered leaf of the Stevia sp. plant. 4.4 g/L. Eight weeks after the treatment, muscle tension recording, and measurement of oxidative stress markers were performed: levels of reactive oxygen species (ROS) and catalase activity. Additionally, body weight, postprandial glucose, and food intake were recorded throughout the experiment. The SRA caused an increase in body weight and a significant reduction in the resistance time to muscle fatigue and the maximum and total muscle tension force. Treatment with SRN caused significant improvements in the parameters studied (p <0.05). We conclude that natural S. rebaudiana is an essential alternative for weight control and the development of antioxidant defense against muscle fatigue but not in synergy with artificial sweeteners.