The flux of various substrates across the ovine fetal and maternal hindlimbs was measured in the fed state and after 5 days of maternal fasting. Whole blood concentrations of glucose, oxygen, ammonia, and six amino acids (glutamate, glutamine, alanine, valine, isoleucine, and leucine) were determined in the fetal and maternal femoral artery and distal inferior vena cava in 15 chronic animal preparations. During fasting the fetal arterial glucose concentration fell by 40% (from 0.828 to 0.494 mM), and the arteriovenous concentration difference decreased by 30% (from 0.148 to 0.099 mM). Similar changes were noted in maternal blood. Fetal oxygen concentrations remained similar between the fed and fasted state, and the fetal arteriovenous oxygen concentration difference increased slightly from 0.861 to 1.02 mM. The glucose oxygen quotient decreased in the fetus from 1.20 to 0.621. In addition, significant changes occurred in the net balance of several amino acids during the fasted state. Both alanine and glutamine, which demonstrated a positive uptake by the fetal hindlimb during the fed state, showed a substantial efflux from the fetal hindlimb during the fasting period. The fetal arteriovenous concentration difference of the branched-chain amino acids (leucine, isoleucine, and valine) increased significantly during fasting. These observations are consistent with the hypothesis that the ovine fetus adapts to a diminished supply of glucose from the mother by enhanced amino acid catabolism and, possibly, proteolysis with subsequent release of gluconeogenic precursors in the form of alanine and glutamine.