We used transdiaphragmatic peak twitch tension (PTT) elicited by bilateral phrenic nerve stimulation to ascertain whether low-frequency (LF) diaphragmatic fatigue (DF) can be induced in spontaneously breathing humans by a combination of an inspiratory resistive load (IRL) and graded treadmill exercise (GXT). Our subjects were 10 young males with normal cardiopulmonary function. Before exercise we measured PTT in each subject by administering supramaximal electrical pulses of 100-microseconds duration at a frequency of 1 Hz to each phrenic nerve with the subjects breath holding at functional residual capacity at a given thoracoabdominal configuration. A minimum of six satisfactory PTT measurements were made in each subject, and we computed the 95% confidence limits (CL) for each subject. The subjects then inspired through a resistive load of 38 cmH2O.1(-1).s-1 while carrying out the GXT until exhaustion. After the GXT, PTT was remeasured in all subjects. In five of the subjects, the post-GXT mean PTT fell below the 95% CL of the pre-GXT mean PTT. However, post-GXT PTT means for the other five subjects were within the 95% CL of the pre-GXT means. In conclusion, using PTT as a measure of LFDF, these results demonstrate that LFDF can be produced in 50% of spontaneously breathing young normal males.