INTRODUCTION: In this study, we examined the acute effects of a short video-based body scan mindfulness practice on the heart rate variability (HRV) and cognitive performance of professional female basketball players after the first half of a simulated basketball game. METHODS: In this crossover randomized controlled trial, nine professional athletes completed a physical loading protocol on two separate days. The protocol consisted of a 10-min Yo-Yo Intermittent Recovery Test Level 1 in the first quarter, followed by a 10-min basketball game in the second quarter. Immediately afterward, they were asked to engage in a 10-min mindfulness practice or watch a 10-min nature-based documentary as a type of mental intervention. Their HRV, Rating of Perceived Exertion (RPE), National Aeronautics and Space Administration Task Load Index 2 (NASA TLX-2), and Go/No-Go test scores were recorded immediately before and after the physical loading and after the mental intervention. RESULTS: The physical demand, effort, and frustration level subscales of the NASA TLX-2 and the RPE scores were found to be significantly higher after the physical loading, and they returned to the baseline level after both types of mental intervention. The Go/No-Go test scores did not differ depending on the measurement time. All time- and frequency-domain heart rate variability parameters, except the low-to-high frequency ratio, were found to be significantly high immediately after the physical loading protocol. However, these parameters returned to their initial levels after both types of mental intervention. DISCUSSION: Completing the tests involved in the study protocol successfully induced physical fatigue, as evidenced by consistent measurement tools, but the one-time and short-term mindfulness practice had no additional benefits for the recovery of heart rate variability, cognitive tasks, or subjective assessment methods, such as RPE and NASA TLX-2, in basketball players with no previous experience of mindfulness practice.