Responses of several folate-metabolizing pathways to dietary folic acid were studied in 2-week-old chicks. Oxidation of a histidine load to carbon dioxide was impaired in folate-deficient chicks. There was a curvilinear relation between oxidation and dietary folate, and maximum oxidation occurred with 2 mg supplemental folic acid/kg. Hepatic activities of glutamic acid formiminotransferase (EC 2.1.2.5) and glycine N-methyltransferase (EC 2.1.1.20) were not affected significantly (P > 0.05) by dietary folic acid. The activity of dihydrofolate reductase (EC 1.5.1.3) in erythrocytes was elevated in folate-deficient chicks. These studies show that the activities of two folate-dependent pathways can be used as biochemical criteria of folate status in chicks.