Propose: the propose of this study was to discover the influence of sand and natural grass on muscle overuse in female rugby players after an induced fatigue test. Methods: the participants of this study were 15 female amateur rugby players (23.4 ± 4.42 years). RSA Test consisted of six-sprints of 40 m (20 + 20 m) and was performed in two different surfaces (natural grass and sand). Before and immediately after completing the RSA, the contractile capacity of the biceps femoris and the rectus femoris of both legs was evaluated through Tensiomyography (TMG). Results : players also did 2 CMJ jumps before and after the RSA to assess the muscle fatigue. CMJ jump high decreased (-2.89 cm; ES= 0.67; IC: to -4.59 to -1.18) after having performed the RSA Test on sand versus natural grass. Rectus femoris presented higher values of Tc (11.66 ms; ES= 1.00; IC: 4.03 to 9.29; p ≤0.01) and Dm (1.20 mm; ES= 0.80; IC: 0.21 to 2.61; p <0.05) on sand than on natural grass after finishing the RSA while the biceps femoris do not display any differences regarding surfaces. Conclusion : therefore, muscular response on rectus femoris after repetitive-sprint-actions differ between different surfaces (sand and natural grass).