Reptiles living in heterogeneous thermal environments are often thought to show behavioural thermoregulation or to become inactive when environmental conditions prevent the achievement of preferred body temperatures. By contrast, thermally homogeneous environments preclude behavioural thermoregulation, and ectotherms inhabiting these environments (particularly fish in which branchial respiration requires body temperature to follow water temperature) modify their biochemical capacities in response to long-term seasonal temperature fluctuations. Reptiles may also be active at seasonally varying body temperatures and could, therefore, gain selective advantages from regulating biochemical capacities. Hence, we tested the hypothesis that a reptile (the American alligator Alligator mississippiensis) that experiences pronounced seasonal fluctuations in body temperature will show seasonal acclimatisation in the activity of its metabolic enzymes. We measured body temperatures of alligators in the wild in winter and summer (N=7 alligators in each season), and we collected muscle samples from wild alligators (N=31 in each season) for analysis of metabolic enzyme activity (lactate dehydrogenase, citrate synthase and cytochrome c oxidase). There were significant differences in mean daily body temperatures between winter (15.66+/-0.43 degrees C; mean +/- S.E.M.) and summer (29.34+/-0.21 degrees C), and daily body temperatures fluctuated significantly more in winter compared with summer. Alligators compensated for lower winter temperatures by increasing enzyme activities, and the activities of cytochrome c oxidase and lactate dehydrogenase were significantly greater in winter compared with summer at all assay temperatures. The activity of citrate synthase was significantly greater in the winter samples at the winter body temperature (15 degrees C) but not at the summer body temperature (30 degrees C). The thermal sensitivity (Q(10)) of mitochondrial enzymes decreased significantly in winter compared with in summer. The activity of mitochondrial enzymes was significantly greater in males than in females, but there were no differences between sexes for lactate dehydrogenase activity. The differences between sexes could be the result of the sex-specific seasonal demands for locomotor performance. Our data indicate that biochemical acclimatisation is important in thermoregulation of reptiles and that it is not sufficient to base conclusions about their thermoregulatory ability entirely on behavioural patterns.