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Purpose: Previous research suggests that the percentage of maximal oxygen uptake attained and the time it is sustained close to maximal oxygen uptake (eg, >90%) can serve as a good criterion to judge the effectiveness of a training stimulus. The aim of this study was to investigate the acute effects of adding vibration during varied high-intensity interval training (HIIT) sessions on physiological and neuromuscular responses. Methods: Twelve well-trained cyclists completed a counterbalanced crossover protocol, wherein 2 identical varied HIIT cycling sessions were performed with and without intermittent vibration to the lower-intensity workloads of the work intervals (6 × 5-min work intervals and 2.5-min active recovery). Each 5-minute work interval consisted of 3 blocks of 40 seconds performed at 100% of maximal aerobic power interspersed with 60-second workload performed at a lower power output, equal to the lactate threshold plus 20% of the difference between lactate threshold and maximal aerobic power. Oxygen uptake and electromyographic activity of lower and upper limbs were recorded during all 5-minute work intervals. Results: Adding vibration induced a longer time ≥90% maximal oxygen uptake (11.14 [7.63] vs 8.82 [6.90] min, d = 0.64, P = .048) and an increase in electromyographic activity of lower and upper limbs during the lower-intensity workloads by 20% (16%) and 34% (43%) (d = 1.09 and 0.83; P = .03 and .015), respectively. Conclusion: Adding vibration during a varied HIIT session increases the physiological demand of the cardiovascular and neuromuscular systems, indicating that this approach can be used to optimize the training stimulus of well-trained cyclists.