Based on studies in Fe-deficient calves demonstrating enhanced blood lactate concentrations during treadmill exercise, the hypothesis was advanced that glucose metabolism is also disturbed at rest. Insulin-dependent glucose metabolism was therefore investigated in calves fed milk replacer containing 20 or 50 mg Fe/kg. Calves receiving only 20 mg Fe/kg of milk replacer developed moderate Fe deficiency anemia and had lower average daily gain than calves fed milk replacer containing 50 mg Fe/kg, but feed intake and feed refusals did not differ between groups. In Fe-deficient calves, insulin responses to glucose, based on intravenous and oral glucose tolerance and hyperglycemic clamp tests, were normal. Using hyperglycemic and euglycemic hyperinsulinemic clamps combined with [13C6]glucose infusions, glucose utilization and tissue sensitivity to insulin were greater in Fe-deficient calves than in Fe-adequate calves. Observed reductions in growth performance may be explained by decreased activity of Fe-dependent enzymes, increased anaerobic glycolysis and lactate-glucose cycling, adaptations that are expected to be energy expensive.