The objective of this study is to verify the coincidence between the occurrence of the electromyographic fatigue threshold (EMGth) and the ventilatory threshold (Vth) in an incremental test in the cyclosimulator, as well as to compare the calculation of the RMS from the EMG signal using different time windows. Thirteen male cyclists (73.7 +/- 12.4 kg and 174.3 +/- 6.2 cm) performed a ramp incremental test (TI) in a cyclosimulator until voluntary exhaustion. Before the start of each TI subjects had the active bipolar electrodes placed over the superficial muscles of the quadriceps femoris (QF) of the right leg: rectus femoris (RF), vastus medialis (VM) and vastus lateralis (VL). The paired student's t test, pearson's correlation coefficient and the analysis method described by Bland and Altman for the determination of the concordance level were used for statistical analysis. The significance level adopted was P < 0.05. Although no significant differences were found between Vth and the EMGth calculated from windows of 2, 5, 10, 30 and 60 seconds in the studied muscles, it is suggested that the EMGth values determined from the calculation of the RMS curve with windows of 5 and 10 seconds seem to be more appropriate for the calculation of the RMS curve and determination of EMGth from visual inspection.