Although there is a growing interest for the effects of intermittent fasting on energy balance, this study aimed to compare appetite, energy intake and food reward responses with an energy depletion induced either by 24-h food restriction or an equivalent deficit with exercise in healthy males. In all, twelve healthy lean males (21·5 (sd 0·5) years old; BMI: 22·5 (sd 1·7) kg/m2) participated in this study. Body composition, aerobic capacity, food preferences and energy intake were assessed. They randomly completed three conditions: (i) no depletion (CON); (ii) full 24-h energy restrictions (Def-EI); and (iii) exercise condition (Def-EX). Ad libitum energy intake and food reward were assessed at the end of each session. Appetite feelings were assessed regularly. Ad libitum energy intake was higher on Def-EI (7330 (sd 2975) kJ (1752 (sd 711) kcal) compared with that on CON (5301 (sd 1205) kJ (1267 (sd 288) kcal)) (P<0·05), with no difference between CON and Def-EX (6238 (sd 1741) kJ (1491 (sd 416) kcal) (P=0·38) and between Def-EX and Def-EI (P=0·22). There was no difference in the percent energy ingested from macronutrients. Hunger was lower on CON and Def-EX compared with Def-EI (P<0·001). Satiety was higher on CON and Def-EI compared with that on Def-EX (P<0·001). There was a significant interaction condition × time for food choice fat bias (P=0·04), showing a greater preference for high-fat v. low-fat food during Def-EI and Def-EX. Although 24-h fasting leads to increased energy intake at the following test meal (without total daily energy intake difference), increased hunger profile and decreased post-meal food choice fat bias, such nutritional responses are not observed after a similar deficit induced by exercise.