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The primary aim of this study was to compare the measured oxygen consumption (Measured -V O 2 ) in a simulated futsal game (S-Game) with the estimated oxygen consumption (Estimated -V O 2 ) through a regression equation between heart rate (HR) and oxygen consumption ( V O 2 ) (HR- V O 2 ) in treadmill running, and a secondary aim was to calculate the total energy expenditure (EE) in S-Game. Ten professional players (22.20 ± 3.22 years) were evaluated. HR- V O 2 was determined individually in the continuous test on the treadmill (Cont Test ). The Measured -V O 2 in S-Game was compared with the Estimated -V O 2 in the Cont Test . Alactic and lactic pathways were estimated by V O 2 . The Estimated -V O 2 presented no statistically significant difference with the Measured -V O 2 , using the paired t -test ( p = 0.38). However, the correlation between Estimated- and Measured- V O 2 was very weak ( r = -0.05), and it presented poor agreement (concordance correlation coefficient = -0.04). In addition, a Bland-Altman plot presented bias of -2.8 ml/kg/min and individual difference as large as 19 ml/kg/min. The HR- V O 2 determined by the Cont Test was not a good individual predictor of V O 2 . The high intensity and intermittent nature of the futsal game possibly caused dissociation in the HR- V O 2 relationship. Cont Test is not recommended for estimating V O 2 and calculating individual EE in the futsal game. This is recommended only for the group mean. The total EE in S-Game was 13.10 ± 1.25 kcal.min -1 (10.81 ± 1.57 metabolic equivalents). The contributions from the metabolic pathways were as follows: aerobic (93%), alactic (5%), and lactic (2%)., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Silva, Nakamura, Papoti, da Silva and Dos-Santos.)