Aim: The objective of this current research was to compare fat oxidation rates during an incremental cycling exercise test in a temperate vs. hot environment.Methods: Twelve healthy young participants were recruited for a randomised crossover experimental design. Each participant performed a VO2max test in a thermoneutral environment followed by two cycling ramp test trials, one in a temperate environment (18.3°C) and another in a hot environment (36.3°C). The ramp test consisted of 3-min stages of increasing intensity (+10% of VO2max) while gas exchange, heart rate and perceived exertion were measured.Results: During exercise, there was a main effect of the environment temperature on fat oxidation rate (F = 9.35, P = 0.014). The rate of fat oxidation was lower in the heat at 30% VO2max (0.42 ± 0.15 vs.0.37 ± 0.13 g/min; P = 0.042), 60% VO2max (0.37 ± 0.27 vs.0.23 ± 0.23 g/min; P = 0.018) and 70% VO2max (0.22 ± 0.26 vs.0.12 ± 0.26 g/min; P = 0.007). In addition, there was a tendency for a lower maximal fat oxidation rate in the heat (0.55 ± 0.2 vs.0.48 ± 0.2 g/min; P = 0.052) and it occurred at a lower exercise intensity (44 ± 14 vs.38% ± 8% VO2max; P = 0.004). The total amount of fat oxidised was lower in the heat (5.8 ± 2.6 vs 4.6 ± 2.8 g; P = 0.002). The ambient temperature also produced main effects on heart rate (F = 15.18, P = 0.005) and tympanic temperature (F = 25.23, P = 0.001) with no effect on energy expenditure (F = 0.01, P = 0.945).Conclusion: A hot environment notably reduced fat oxidation rates during a ramp exercise test. Exercising in the heat should not be recommended for those individuals seeking to increase fat oxidation during exercise.