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In a crossover study, either a placebo paste or N,N-dimethylglycine was administered orally at a dose rate of 1.2 mg/kg twice daily for five days to six thoroughbred horses, with bodyweights ranging from 424 to 492 kg. Using previously determined regression equations for oxygen uptake (VO2) against speed for each horse, a standardised exercise test was given with speeds equivalent to fixed percentages of the maximum oxygen uptake (VO2max). The test consisted of two minutes at speeds equivalent to approximately 40 per cent and 50 per cent VO2max, and one minute at speeds that produced approximately 60, 70, 80, 90 and 100 per cent VO2max. During the last five seconds of each exercise stage, the values of VO2, carbon dioxide production (VCO2), heart rate, arterial blood and plasma lactate concentrations, arterial blood gases and pH were measured. Before and immediately after the exercise test, muscle biopsies were collected from the middle gluteal muscle to determine the muscle lactate concentrations. The administration of N,N-dimethylglycine produced no significant differences in any of the measured values, and it is concluded that the compound has no beneficial effects on cardiorespiratory function or lactate production in the exercising horse.