The purpose of the study is to investigate the effect of acute bouts of high-intensity aerobic exercise of differing durations on subjective appetite, food intake and appetite-associated hormones in endurance-trained males. Twelve endurance-trained males (age = 21 ± 2 years; BMI = 21.0 ± 1.6 kg/m2; VO2max = 61.6 ± 6.0 mL/kg/min) completed four trials, within a maximum 28 day period, in a counterbalanced order: resting (REST); 15 min exercise bout (15-min); 30 min exercise bout (30-min) and 45 min exercise bout (45-min). All exercise was completed on a cycle ergometer at an intensity of ~76% VO2max. Sixty minutes post exercise, participants consumed an ad libitum meal. Measures of subjective appetite and blood samples were obtained throughout the morning, with plasma analyzed for acylated ghrelin, total polypeptide tyrosine-tyrosine (PYY) and total glucagon-like peptide 1 (GLP-1) concentrations. The following results were obtained: Neither subjective appetite nor absolute food intake differed between trials. Relative energy intake (intake – expenditure) was significantly greater after REST (2641 ± 1616 kJ) compared with both 30-min (1039 ± 1520 kJ) and 45-min (260 ± 1731 kJ), and significantly greater after 15-min (2699 ± 1239 kJ) compared with 45-min (condition main effect, P < 0.001). GLP-1 concentration increased immediately post exercise in 30-min and 45-min, respectively (condition × time interaction, P < 0.001). Acylated ghrelin was transiently suppressed in all exercise trials (condition × time interaction, P = 0.011); the greatest, most enduring suppression, was observed in 45-min. PYY concentration was unchanged with exercise. In conclusion, high-intensity aerobic cycling lasting up to 45 min did not suppress subjective appetite or affect absolute food intake, but did reduce relative energy intake, in well-trained endurance athletes. Findings question the role of appetite hormones in regulating subjective appetite in the acute post-exercise period.