Dobbin, N, Lamb, KL, and Twist, C. Selected physiological, perceptual, and physical performance changes during two bouts of prolonged high-intensity intermittent running separated by 72 hours. J Strength Cond Res 31(12): 3474–3481, 2017—This study investigated the effects of performing a second 90-minute intermittent running protocol 72 hours after an initial trial on selected physiological, perceptual, and sprint running measures. Eight subelite soccer players provided measures of isokinetic muscle function, countermovement jump (CMJ), 10-m sprinting, and muscle soreness before, and at 0, 24, 48, and 72 hours after a 90-minute intermittent high-intensity running bout (IHIR-1). A second 90-minute IHIR bout (IHIR-2) was performed 72 hours after the first. Heart rates, ratings of perceived exertion (RPE), blood lactate concentration [Bla], and 10-m sprint times were recorded periodically during both IHIR. Analysis of effects revealed that in the 72-hour period after IHIR-1, there were most likely increases in muscle soreness and likely to very likely deteriorations in CMJ, 10-m sprint, and isokinetic muscle function. During IHIR-2, heart rates (possibly to likely) and [Bla] (possibly to very likely) were lower than IHIR-1, whereas RPE remained unchanged. Sprint times during IHIR-2 were also likely to very likely higher than in IHIR-1. It was evident that these team sport players exposed to repeat bouts of prolonged high-intensity running within 72 hours downregulated their sprint performances in the second bout despite no change in perceived effort. These findings have implications for managing training and match loads during periods of intense scheduling.