Elite endurance athletes possess a high capacity for whole-body maximal fat oxidation (MFO). The aim was to investigate the determinants of a high MFO in endurance athletes. The hypotheses were that augmented MFO in endurance athletes is related to concomitantly increments of skeletal muscle mitochondrial volume density (Mito ) and mitochondrial fatty acid oxidation (FAO ), that is, quantitative mitochondrial adaptations as well as intrinsic FAO per mitochondria, that is, qualitative adaptations. Eight competitive male cross-country skiers and eight untrained controls were compared in the study. A graded exercise test was performed to determine MFO, the intensity where MFO occurs (Fat ), and . Skeletal muscle biopsies were obtained to determine Mito (electron microscopy), FAO , and OXPHOS (high-resolution respirometry). The following were higher (P < 0.05) in endurance athletes compared to controls: MFO (mean [95% confidence intervals]) (0.60 g/min [0.50-0.70] vs 0.32 [0.24-0.39]), Fat (46% [44-47] vs 35 [34-37]), (71 mL/min/kg [69-72] vs 48 [47-49]), Mito (7.8% [7.2-8.5] vs 6.0 [5.3-6.8]), FAO (34 pmol/s/mg muscle ww [27-40] vs 21 [17-25]), and OXPHOS (108 pmol/s/mg muscle ww [104-112] vs 69 [68-71]). Intrinsic FAO (4.0 pmol/s/mg muscle w.w/Mito [2.7-5.3] vs 3.3 [2.7-3.9]) and OXPHOS (14 pmol/s/mg muscle ww/Mito [13-15] vs 11 [10-13]) were, however, similar in the endurance athletes and untrained controls. MFO and Mito correlated (r = 0.504, P < 0.05) in the endurance athletes. A strong correlation between Mito and MFO suggests that expansion of Mito might be rate-limiting for MFO in the endurance athletes. In contrast, intrinsic mitochondrial changes were not associated with augmented MFO.