The relationship between body size and the depletion of white muscle metabolites (e.g., PCr, ATP, glucose and glycogen) was examined in two sizes (30 or 700 g) of rainbow trout deprived of food for one, four or seven days at either 5 or 15 °C. Following 7 days of food deprivation at 15 °C, the levels of muscle glycogen decreased by approximately 50% in small fish relative to control values (i.e., day 1). In comparison, small fish acclimated to cold temperatures did not exhibit a significant reduction of muscle glycogen over the seven day fasting regime. In contrast to small fish, the levels of white muscle glycogen in large fish remained unchanged after food deprivation, regardless of acclimation temperature. A seven day deprivation of food also resulted in a 50% depletion of white muscle glucose concentrations in small and large fish acclimated to warm temperatures, but there were no significant changes in this variable in fish acclimated to cold temperatures. In contrast to the negative effects of food deprivation on white muscle glycogen and glucose levels, the concentrations of white muscle PCr and ATP were not greatly affected by food deprivation under any of the experimental conditions. Taken together, these results clearly show that food deprivation can have an important influence on the storage of energy metabolites for anaerobic energy production, particularly in small fish at warm temperatures. In the future, it may be very important to consider the physiological effects of short-term food deprivation when interpreting results from studies in which fish have been fasted prior to treatments such as exercise.