Competitive sports involve physiological, technical and psychological skills, which influence directly on individuals’ performance. This study aims to investigate the levels of perceived stress and Heart Rate Variability before and after matches with victory and defeat in professional eSports athletes. Our hypothesis was that the winners would have better autonomic and stress responses after match, thus corroborating the literature on neurocardiac connections. Fifty male eSport players were selected players from 10 different Brazilian teams. The experiment was carried out in 2 (...) sessions. Firstly, after signing the informed consent form, 24 h before the game, anthropometric, physical activity levels and time of expertise data were recorded only for sample characterization and the players were familiarized with the perceived stress scale—10 and the HRV measurements. Secondly, players performed the PSS-10 and HRV recording at rest by 10 min 60 and 30 min before the game and 10 min after the end of the game. Overall, concerning PSS-10 our findings show that VG had significant reduced scores in post-game time compared to baseline and pre-game times, while DG had significant increased scores in post-game time compared to BL and pre-game times. Regarding HRV, our results demonstrate that VG had significant increase in RR, SDNN, rMSSD, pNN50 and HF, and significant decrease in LF and LF/HF, while DG had a significant decrease in RR, SDNN, rMSSD and HF, and significant increase in LF and LF/HF. It was observed that VG had better HRV responses as well as lower levels of perceived stress, while DG had worst HRV responses and higher levels of perceived stress. (shrink)