On the straight line, more horses were subjectively declared sound at higher speeds, whilst different objective asymmetry measures showed only slight and inconsistent changes. On the circle, speed had no significant effect on the subjective assessment, with an increase in objectively measured asymmetry at higher speeds possibly balanced by a decrease in sensitivity of the observers for this asymmetry. Horses visually examined for subtle lameness on the straight should therefore be evaluated at a slow speed. Trotting speed should be consistent on repeated occasions, especially during objective gait analysis on the circle, to avoid the interaction of treatment effects and speed effects.