The activities of acetoacetyl-CoA thiolase, hydroxymethylglutaryl-CoA synthase and lyase and acetoacetyl-CoA deacylase were measured in homogenates of samples of liver, rumen epithelium (long papillae), kidney and lactating mammary gland derived from slaughtered cows. 2. The activities of the four enzymes in bovine liver were similar to the activities previously reported for the corresponding enzymes in rat liver. 3. Acetoacetyl-CoA thiolase and hydroxymethylglutaryl-CoA synthase and lyase were present in rumen epithelium. The activities of the enzymes were all lower on a wet weight basis than in liver. Only very slight deacylase activity was detected. 4. Kidney contained acetoacetyl-CoA thiolase, hydroxymethylglutaryl-CoA lyase and acetoacetyl-CoA deacylase, but only trace amounts of hydroxymethylglutaryl-CoA synthase. 5. Mammary gland contained acetoacetyl-CoA thiolase and some hydroxymethylglutaryl-CoA lyase, but virtually no hydroxymethylglutaryl-CoA synthase or acetoacetyl-CoA deacylase. 6. Since physiologically significant ketogenesis probably occurs solely via the hydroxymethylglutaryl-CoA pathway, it is evident that, of the four tissues examined, such ketogenesis must be restricted to the liver and the rumen epithelium. 7. All the enzymes except hydroxymethylglutaryl-CoA lyase were also assayed in the four tissues derived from cows suffering from bovine lactational ketosis. Ketosis did not cause a statistically significant change in the activity of any of the enzymes measured. 8. Hepatic hydroxymethylglutaryl-CoA synthase and lyase were found to be associated mainly with the particulate fraction, as in the rat. A considerably greater proportion of these enzymes was found to be present in the cytoplasmic fraction from rumen epithelium, although it was not excluded that this was due to mitochondrial damage during homogenization. 9. Appreciable hydroxymethylglutaryl-CoA synthase was also present in epithelium from the dorsal region of the rumen, from the reticulum and from the omasum, but not from the abomasum.
