Experiments were designed to produce biochemical changes in both serum and bone in rats by long-term dietary deficiencies of manganese (Mn) and copper (Cu). Weanling rats were fed one of three diets: 1) normal Mn and Cu (N), 2) low Mn and Cu (L) and 3) depleted Mn and normal Cu (D). After 12 mo, serum Ca and P were significantly greater in the D rats than the N rats (134, 50 mg/L and 103, 39 mg/L, respectively). Serum and femur Mn levels were lower in D rats than in normal rats. Serum and femur Cu levels were lower in L rats than N rats. Femur Ca concentrations were significantly lower in D rats than in N rats (180 and 272 mg/g, respectively). Radiographic differences could be detected in some rats fed the L and D diets. In the two deficient groups, increased serum Ca was negatively correlated with bone Ca (r = -0.69; P less than 0.01). This biochemical association may represent alterations in regulatory control of Ca at the level of the bone (decreased mineralization) combined with an increase in bone resorption. The effect of long-term dietary deficiencies in Mn and Cu should be considered in human bone metabolism.