The purposes of the present study were two fold: (1) to determine if the model used for estimating the physical working capacity at the fatigue threshold (PWCFT) from electromyographic (EMG) amplitude data during incremental cycle ergometry could be applied to treadmill running to derive a new neuromuscular fatigue threshold for running, and (2) to compare the running velocities associated with the PWCFT, ventilatory threshold (VT), and respiratory compensation point (RCP). Fifteen college-aged subjects (21.5 ± 1.3 y, 68.7 ± 10.5 kg, 175.9 ± 6.7 cm) performed an incremental treadmill test to exhaustion with bipolar surface EMG signals recorded from the vastus lateralis. There were significant (p < 0.05) mean differences in running velocities between the VT (11.3 ± 1.3 km h(-1)) and PWCFT (14.0 ± 2.3 km h(-1)), VT and RCP (14.0 ± 1.8 km h(-1)), but not the PWCFT and RCP. The findings of the present study indicated that the PWCFT model could be applied to a single continuous, incremental treadmill test to estimate the maximal running velocity that can be maintained prior to the onset of neuromuscular fatigue. In addition, these findings suggested that the PWCFT, like the RCP, may be used to differentiate the heavy from severe domains of exercise intensity.