In Standardbreds, the main aim of early training (begun during the growth period) is the mechanization of athletes leading to a particular gait called the ‘flying trot’. The present cross-sectional study was undertaken to investigate the biomechanical and physiological factors involved in this gait change, and aimed to analyse the effect of age on gait and energetic variables in a population of Standardbred horses under training. One hundred and forty-three horses aged from two to seven years were tested on a track at three speeds (8.5, 10 and 11.6 m s−1) with a gait-analysis system. Gait variables (temporal and linear variables, symmetry, regularity, two-beat rhythm, dorso-ventral, longitudinal and lateral activities) were compared between four age groups (two-, three-, four-, and five-year-olds and above). After a standardized exercise test, two energetic variables (V4 and V200) were also compared between these groups. Most variables were influenced by age/training status. The results indicated that, from young to mature racehorses, stride length and duration increase; and gait becomes more symmetric and more regular. We also observed a decrease in dorso-ventral, longitudinal and lateral activities, i.e. a decrease of thoracic displacements. These differences could be elucidated at slow speed and were still obvious at high speed. V4 and V200 also increased significantly with age/training status. All of these results indicate an improvement in co-ordination and a decrease in metabolic cost with increasing age/training status. Thus gait acquisition could be related to an improvement in trotting efficiency.