To establish the effect of the locomotor pattern on heart rate (HR) and plasma lactate (LA), 28 horses, 18 Andalusian (AN) and 10 Anglo-Arabian (AA), aged 3-4 years, were studied. After a warm-up period, the horses performed a four-step test at 5, 6, 7 and 8 m/s, covering 1000 m at each velocity. HR was monitored, LA was analysed at rest and after each workload, and images were filmed. The locomotor parameters determined were stride duration (SD), frequency (SF) and length (SL), step and bipedal support durations, stance (restraint and propulsion) and swing phase durations, and stride vertical component. The HR and LA were significantly higher in the AN horses from velocities of 5 m/s. Similarly, the stride vertical component was higher in the AN horses at the trot and in the leading forelimbs at the gallop. Conversely, at all the galloping velocities, swing phase duration and stride length were longer in the AA horses. Significant correlations between HR, LA and locomotor pattern were only found in the AN horses. It was concluded that the greater stride vertical component in the AN horses limits SL and the cranial advancement of the hoof, with the result that reaching a longer SL triggers an increase in HR and LA. AA horses reach a balance between SL and SF, improving the efficiency of the gait.