There is limited information about the effect of different levels of competition on the physical fitness of college soccer players. Therefore, we compared the physical fitness of soccer players from different levels of competition within the same college team. We tested 111 male college soccer players from the same college team for high (first), moderate (second), and low (third) competition levels. We used the Yo-Yo Intermittent Recovery Test level 2 (Yo-Yo IR2); 10-, 20-, and 30-m sprints; countermovement jump (CMJ); and 5 rebound jump test (5RJ). The Yo-Yo IR2 and 5RJ results were significantly higher in the first group than in the second and third groups. Further, the CMJ was significantly higher in the first group than in the third group. Between the second and third groups, the Yo-Yo IR2, CMJ, and 5RJ results were significantly higher in the second group than in the third group. However, the third group had a significantly higher 10-m sprint time than the second group. These findings suggest that college soccer players from the high level of competition had superior values for the Yo-Yo IR2, CMJ, and 5RJ tests than the players from the low level of competition within the same team. In contrast, the sprint test did not reveal a superior performance for players from the high level of competition. Therefore, jump tests may serve as an index of anaerobic power that better reflects different levels of competition within the same team compared with sprint tests.