SummaryThe objectives of this study were to: 1) determine the effects of heat and high relative humidity (RH) on the clinical and physiological responses of horses during and after daily exercise training and 2) determine whether repeated exposure to, and exercise in, the heat would result in improved thermal tolerance (heat acclimation). Six trained Thoroughbred horses completed 1 h of submaximal exercise in cool, dry conditions (CD) and during a daily 4 h period of exposure to high heat and humidity (HH, room temperature = 33–35°C, RH = 80–85%) for 22 days. Rectal temperature (Tre) and heart rate (HR) were measured before, during and after exercise and respiratory rate (RR) was measured before exercise and during a 2 h recovery. In HH, the rate of rise in Trewas significantly higher than in CD. However, by HH Day 5, Trebefore, during and after exercise was significantly lower than on HH Day 1. The day‐to‐day decrease in Treduring exercise was reflected in significant decreases in heat storage following exercise by HH Day 10 (910 ± 47 kcal) when compared to HH Day 1 (1211 ± 75 kcal). At rest, RR was initially higher in HH than CD, and a further increase in pre‐exercise RR from HH Day 1 to Day 10 may have contributed to the lower pre‐exercise Tre. Recovery RR was higher after HH Day 1 and was associated with a lower end‐of‐exercise Treand reduced heat storage by HH Day 5. Pre‐exercise HR did not change in the 1 h before exercise in CD and did not differ from HH Days 1–22. By HH Day 10, mean HR during the latter part of exercise was lower than HH Day 1 and was not different from pre‐exercise by 60 min of recovery. Pre‐exercise body mass did not change during the 3 wk period, and the decrease in body mass that occurred during the 4 h training period was significantly attenuated by HH Day 15 (9.8 ± 0.8 kg) when compared to HH Day 1 (12.5 ± 0.8 kg). Over the 3 week period of HH, mean 24 h water consumption increased from 26.0 ± 2.1 litres to 39.5 ± 3.2 litres, largely reflecting a 2‐fold increase in water intake during the 4 h period of heat exposure. It is concluded that 3 weeks of daily exposure to, and exercise in, hot and humid ambient conditions resulted in a progressive reduction in thermal and cardiovascular strain. Furthermore, the reported physiological adaptations are consistent with an improved ther