The changes in long-chain fatty acid oxidation during the first 24 h after birth were studied in isolated rabbit hepatocytes and liver mitochondria. The eightfold increase in this oxidation which occurs in hepatocytes between birth and 24 h was not triggered by a concomitant decrease in long-chain fatty acid esterification. Indeed, in isolated hepatocytes from 24-h-old rabbits, the 75% inhibition of the oxidation by 2-tetradecylglycidic acid, resulted in a total redirection of oleate metabolized towards triacylglycerol synthesis. Polarographic measurements of mitochondrial respiration showed that oxidative phosphorylation and respiratory chain capacity were fully functional at birth. By contrast, in liver mitochondria isolated from newborn rabbits, the rate of oxygen consumption from palmitoyl-L-carnitine was 60% higher than from palmitoyl-CoA. Similarly palmitoyl-CoA oxidation was increased 1.5-fold in isolated mitochondria from 24-h-old rabbits. These results were in agreement with the twofold increase in the activity of hepatic carnitine palmitoyltransferase I between birth and 24 h. However it is unlikely that the twofold increase in this enzyme activity totally explained the eightfold increase in long-chain fatty acid oxidation in isolated newborn rabbit hepatocytes. It was shown that the rate of the oxidation in isolated hepatocytes was inversely related to the rate of lipogenesis. Nevertheless, malonyl-CoA concentration per se is probably not the factor involved in the regulation of the oxidation between birth and 24 h, since a 90% decrease in hepatic malonyl-CoA concentration was not associated with a stimulation of long-chain fatty acid oxidation. The more likely mechanism was the 30-fold decrease in the sensitivity of carnitine palmitoyltransferase I to malonyl-CoA inhibition.