1. The effects of riboflavin deficiency on growth, whole-body oxygen consumption, cytochrome c oxidase (EC 1.9.3.1) activity and GDP-binding capacity of brown adipose tissue were measured in three groups of rats: sucking pups, weanling rats, and dams. Control groups were weight-matched, pair-fed or fed ad lib. 2. Riboflavin deficiency reduced growth rate and increased the activation coefficient of erythrocyte glutathione reductase (NAD(P)H) (EC 1.6.4.2), as predicted. In sucking pups it also reduced whole-body O2 consumption per unit body-weight, especially after noradrenaline stimulation. In weanling rats, however, it increased O2 consumption both before and after noradrenaline stimulation. 3. Cytochrome c oxidase (EC 1.9.9.1) activity of brown adipose tissue was not consistently affected by riboflavin deficiency. Binding of [3H]GDP to the mitochondria was increased in the deficient weanling rats. 4. Weanling rats therefore, seemed better able to withstand the effects of severe depletion. Their reduced growth and increased non-shivering thermogenesis helped to counteract the unfavourable ratio of riboflavin:other tissue-building materials. The relevance for thermoregulation in riboflavin-deficient children is discussed.