Purpose <p>The aim of the study is to assess the effect of a running fatigue protocol on static (SB) and dynamic balance (DB) abilities using Wii balance board (WBB) and Y-balance (YB) tests, respectively, in young male recreational athletes.</p> Materials and Methods <p>Participants were randomized into control and experimental groups. They were examined on static and dynamic balance (pre- &amp; post-tests). Then, the experimental group underwent a fatigue protocol, in which speed and incline increased one unit for each passing minute, and stopped when rate of perceived exertion level reached ≥ 18, and the plasma lactate level differed from the baseline by ≥ 4mmol/L.</p> Results <p>Forty male athletes aged between 18–22 years old participated with 20 participants in each group. Between groups analysis showed that the control group scored significantly higher score on YB, however, the experimental group scored significantly lower on WBB in all conditions. Regression analysis revealed that, for every one unit increase in SB score, plasma lactate levels decrease by 0.051 (5.1%).</p> Conclusions <p>Reduced performance on both static and dynamic balance tests after a treadmill running fatigue protocol was observed in recreational young male athletes, compared to a randomly-assigned control group of their peers.</p> Trial registration <p>ClinicalTrials.gov ID: NCT06901219, Retrospectively registered on 16/3/2025.</p>