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Semimembranosus (SM) muscle as well as Semitendinosus-Gracilis (STG) tendon have the same role in knee flexion and tibial internal rotation. Because STG tendons are generally used for anterior cruciate ligament (ACL) reconstruction, some compensational changes in SM muscle might have been induced. The purpose of this study was to investigate the changes of SM muscle affected by harvesting STG tendons. 10 Wistar-strain male rats were divided into control (C) and STG-dissected (STG) groups. Left STG tendons including the distal half of muscle portions were dissected in group STG and only skin incision was performed in group C. 4 weeks after the treatments, fiber types classification and ultrastructural observations were performed. In group STG the decrease of type IIa (fast-twitch fiber with high oxidative capacity) was observed in deep layers of SM muscle (p<0.01). In ultrastructural observations, the increase in lipid droplets and mitochondria and the irregularity of Z disc were observed in deep layers. These morphological changes indicated that the mechanical loading might increase in SM muscle after harvesting of STG. Because of minor injuries in SM muscle, hamstring strength exercise at early stage of rehabilitation program should be carefully performed following ACL reconstruction using STG tendons in clinical practice.