The purpose of this study was to determine the independent and combined moderating effect of aerobic fitness and environmental conditions on physical workloads during collegiate female soccer matches. Nineteen National Collegiate Athletic Association female soccer athletes were included in this study (mean ± SD: age, 20.6 ± 1.4 years; height, 169 ± 6.1 cm; body mass 64.7 ± 5.3 kg). Maximal oxygen consumption (V[Combining Dot Above]O2max) was estimated from the yo-yo intermittent recovery test before preseason training and wet-bulb globe temperature (WBGT) was recorded onsite for home matches and at the nearest weather station for away matches. Relative distance (TD), relative high-speed running distance (%HSD), and relative high metabolic load (%HML) performance were collected during each match using a global positioning system unit (Viper Pod; STATSports, Chicago, IL). Statistically significant differences were observed in TD between LOW WBGT and MOD WBGT (mean difference [MD] = 7.08 m·min; effect size [ES] = 0.54; p < 0.001), in %HSD between LOW WBGT and MOD WBGT (MD = 1.97%; ES = 0.64; p = 0.01) and between LOW WBGT and HIGH WBGT (MD = 2.71%; ES = 1.01; p < 0.001), and in %HML between LOW WBGT and MOD WBGT (MD = 1.24%; ES = 0.56; p < 0.001) and between LOW WBGT and HIGH WBGT (MD = 1.55%; ES = 0.78; p = 0.01). There was a significant interaction between WBGT and V[Combining Dot Above]O2max for %HSD (p = 0.03). These findings demonstrate that physical performance metrics were affected by increased WBGT. In addition, aerobic fitness seemed to moderate the effect of increasing WBGT on %HSD, meaning maximizing aerobic capacity is important for optimizing running performance in the heat. Coaches and sports medicine staff could alter training time and session length based on environmental conditions as well as potentially use aggressive cooling strategies to mitigate the imposed heat stress and decrements in physical performance.