The purpose of this study was to compare the velocity-dependent changes in stride length and stride frequency of horses performing the same incremental exercise test first on a track with a rider and then on a treadmill with a 0 and 3.5% slope successively. Seven French saddle horses undertook the exercise test which consisted of 5 stages of increasing speed for 3 min each with 1 min rest between each stage (1.6, 4.2, 8.3, 9.2 and 10 m/s). The horses were recorded by use of a video camera, and the tapes were analysed with regard to gait parameters. At 1.6, 8.3, 9.2 and 10 m/s, stride frequencies were significantly (p < 0.01) greater on the track than on the treadmill. At the walk, stride length was significantly (p < 0.01) shorter on the track than on the treadmill. There were strong linear relationships between stride length and speed in all experimental conditions (R2 > 0.96). The comparison of the regression coefficients revealed significant differences (p < 0.01) between track and treadmill locomotion; stride length was longer on both the horizontal and inclined treadmill than on the track. The incline of the treadmill did not significantly (p > 0.01) influence stride parameters.