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Trail running involves off-road running over different surfaces of positive and negative unevenness. Given these particularities and the associated physical demands, it is essential to understand this relationship and how fitness levels influence performance. This study aimed to analyze fitness level variations during different times of the season and establish a relationship between changes in fitness levels and accumulated load. Twenty-five trail running athletes (age 36.23 ± 8.30 years) were monitored over 52 weeks. Three periods of