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This study aimed to analyze changes in performance, muscle function, and stress-related biochemical markers in professional soccer players (n = 14) at 4 timepoints (3 for performance and 4 for stress-related biochemical markers) during the soccer season preseason (E1), midseason (E2), end of the season (E3) and after the end of the recovery period (E4). Performance in 5- and 30-m sprints, countermovement jump, and agility, and maximal isokinetic knee extension and knee flexion strength were measured (E1 to E3). We observed increased in-season levels of myoglobin (E2 > E1 and E4; p < 0.05), a higher testosterone/cortisol ratio (T/C), and increased levels of creatine kinase (CK), C-reactive protein, superoxide dismutase (SOD), protein sulfhydryls (–SH), and malondialdehyde (E2 and E3 > E1 and E4; p < 0.05). Lower cortisol concentrations (E3 < E1 and E4; p < 0.05) and glutathione reductase activity (E3 …