The effects of prolonged variable-intensity and short-term high-intensity exercise on indices of oxidative stress and iron homeostasis were compared in six fit horses under cool [20 degrees C, 40 degrees C relative humidity (RH)] or hot/humid (30 degrees C, 80% RHS environmental conditions. The exercise protocols were designed to simulate equine competition, including racing (intense exercise) or the speed and endurance phase of a 3-day event (prolonged exercise). Increased plasma concentrations of lipid hydroperoxides and haemolysate concentrations of oxidised glutathione (GSSG) were measured within 30 min of the completion of exercise, indicating production of reactive oxygen species (ROS) and lipid membrane peroxidation. The horses were unable to complete the prolonged exercise protocol at high temperature and humidity. This coincided with higher maximal values of lipid hydroperoxides [138.2 (17.7) mu M and GSSG [110.6 (18.2) mu M], compared to high-intensity [105.2 (14.9) mu M and 63.6 (8.6) mu M. respectively] or prolonged [100.7 (187) mu M and 86.2 (9.1) mu M, respectively] exercise performed under cooler environmental conditions. Significant correlations were found between the duration of the final stage of exercise during hot/humid environmental conditions and increased levels of lipid hydroperoxides (r=0.85), GSSG (r=0.94), xanthine (r=0.92) and uric acid (r=0.96). Excerise also decreased the iron (Fe)-binding antioxidant activity of the plasma and increased the total plasma Fe levels, although this was only significant for prolonged exorcise in ambient conditions. There was no detectable free Fe in the plasma at any stage of exercise, Other changes in biochemical parameters had returned to pre-exercise levels within 24 h after exercise, The results show that exercise can induce changes in biochemical parameters that are indicative of oxidative stress in the fit horse and that this was, exacerbated during exercise at high temperature and humidity.