The purpose of this study was to determine whether oral L-carnitine supplementation enhances the responses of skeletal muscle to training in seven 2-year-old Standardbreds. Four horses were supplemented with 10 g/day L-carnitine for 10 weeks and 3 horses served as controls. All horses were exercised regularly every second day on a treadmill for 5 weeks (training period) and housed in individual boxes for 5 additional weeks (detraining period). The training period consisted of 8 high- and 8 low-speed exercises carried out in alternating sequence. Gluteus medius muscle biopsies were taken at Weeks 0 (pretraining), 5 (post-training) and 10 (detraining). Muscular adaptations to training were observed mainly in the L-carnitine-supplemented horses and included an increase in the percentage of type IIA fibres (δ35%, P<0.05), atrophy of type I fibres (δ24%, P<0.01), a rise in the capillary-to-fibre ratio (δ40%, P<0.01) and an increase in the quantitative reaction of periodic acid Schiff stain (δ11%, P<0.05), used as an indicator of intrafibre glycogen content. After detraining, most of these adaptations reverted towards the pretraining situation. Therefore, exogenous carnitine has an additive effect on muscular responses to training and this should be favourable to improve athletic performance. Nevertheless, further studies are necessary to show whether muscle carnitine content is a limiting factor for fatty acid oxidation.