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OBJECTIVE: To monitor ultraviolet B light exposure in human milk-fed infants both with and without supplemental vitamin D2, and to measure longitudinally the bone mineral content, growth, and serum concentrations of calcium, phosphorus, 25-hydroxyvitamin D3, 25-hydroxyvitamin D2, 1,25-dihydroxyvitamin D, and parathyroid hormone. DESIGN: Longitudinal, randomized, double-blind, placebo-controlled study of 6 months' duration. SETTING: Patients from private pediatric practice, Madison, Wisconsin. PATIENTS: Sequential sampling of 46 human milk-fed white infants; 24 received 400 IU/day of vitamin D2, and 22 received placebo. An additional 12 patients were followed who received standard infant formula. Eighty-three percent of patients completed a full 6 months of the study. MEASUREMENTS AND RESULTS: Ultraviolet B light exposure and measurements of growth did not differ between groups. At 6 months, the human milk groups did not differ significantly in bone mineral content or serum concentrations of parathyroid hormone or 1,25-dihydroxyvitamin D, although total 25-hydroxyvitamin D values were significantly less in the unsupplemented human milk group (23.53 +/- 9.94 vs 36.96 +/- 11.86 ng/ml; p less than 0.01). However, 25-hydroxyvitamin D3 serum concentrations were significantly higher in the unsupplemented human milk-fed group compared with the supplemented group (21.77 +/- 9.73 vs 11.74 +/- 10.27 ng/ml, p less than 0.01) by 6 months of age. CONCLUSION: Unsupplemented, human milk-fed infants had no evidence of vitamin D deficiency during the first 6 months of life.

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Studies at the University of Wisconsin have demonstrated that photon absorptiometry is a precise, accurate, and reproducible technique for measuring bone mineral content in premature infants and can be used to establish an "intrauterine" curve of bone mineralization in the fetus. Photon absorptiometry can also be used to measure bone width, thereby documenting appositional bone growth. The bone mineral content/bone width ratio may be helpful in identifying disorders of bone mineral metabolism in premature infants. The technique has been used to demonstrate that relatively poor bone mineralization (compared with the intrauterine curve) occurs in very low birth weight infants after birth, regardless of the type of feeding or the presence or absence of bronchopulmonary dysplasia.

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"Preterm" human milk fortified with protein (0.85 gm/dL), calcium (90 mg/dL), and phosphorus (45 mg/dL) was compared with unfortified preterm human milk as a feeding for low birth weight infants. Additionally, a special formula for low birth weight infants (Similac Special Care (SC), 20 cal/oz), was compared with a standard 20 cal/oz formula (Similac). Bone mineral content (BMC), as measured by photon absorptiometry, improved in the study groups fed fortified human milk and Similac SC formula during the first 6 weeks of full oral feedings. Even though the intakes of calcium in the groups fed fortified human milk and Similac SC formula approached the intrauterine requirement for Ca during the third trimester of pregnancy (150 mg/kg/d), the values for BMC in these two groups (37 to 39 mg/cm) at the completion of the study were still considerably less than the intrauterine values for radial BMC at 36 to 37 weeks gestational age (72.6 +/- 14.1 mg/cm). Furthermore, the relative phosphorus deficiency (as determined by increased urinary Ca excretion and increased renal tubular reabsorption of phosphate) in the human milk groups occurred with or without supplements of Ca and P. Rate of weight gain in the fortified human milk group was greater than that of the unfortified human milk group and was comparable to that of infants fed Similac SC formula. Rate of weight gain for the unfortified human milk group was similar to that of infants fed Similac formula containing 20 cal/oz. However, none of the four feeding groups exceeded the 50th percentile for weight at the time of discharge (36 to 37 weeks postconceptional age). The results suggest that fortifying preterm human milk with Ca, P, and protein for low birth weight infants will improve bone mineralization and rate of growth to levels comparable to those achieved with a special formula containing high amounts of protein, Ca, and P.

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We report measurements of urinary inorganic sulfate (iSO4) in 38 very low birth weight (VLBW) premature infants receiving various protein intakes in the first 2 months of life. The primary source of urinary iSO4 is the metabolism of amino acids containing sulfur (methionine, cysteine, taurine). It was hypothesized that urinary iSO4 excretion would be increased in VLBW infants fed the relatively high concentrations of protein in mother's own milk (HM), mother's own milk fortified with 0.85 gm/dl bovine whey (fortified HM), and a special formula for premature infants (Similac Special Care, 20 cal/oz), and that urinary iSO4 excretion would correlate with calcium excretion. VLBW premature infants fed HM (protein intake 3.3 gm/kg day) excreted very small amounts of urinary iSO4 compared with infants fed fortified HM (4.5 gm/kg/day protein), Similac SC (2.9 gm/kg/day protein), or Similac (2.7 gm/kg/day protein), all three of which contain bovine whey. Unlike the case in adults, there was no correlation between either total protein intake and urinary calcium excretion or urinary iSO4 excretion. There was, however, a significant correlation between methionine intake and urinary iSO4 excretion (r = 0.48). We speculate that increased urinary iSO4 excretion is indicative of an overload of sulfur-containing amino acids, namely methionine, present in bovine whey protein. The data also support the ability of premature infants to catabolize relatively large quantities of sulfur-containing amino acids after 2 weeks of age.

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The changes in fat concentration and cumulative fat losses that occur during the delivery of human milk using two different continuous infusion systems were compared with the changes in fat concentration during simulated intermittent gavage or bolus feedings. With both mechanical pumps the largest cumulative fat losses and the greatest decreases in fat concentrations occurred at the slowest infusion rates. State of homogenization of the milk generally made little difference in the changes in fat concentration using the syringe pump, whereas homogenizing the milk increased the fat concentration significantly with the roller pump. With the syringe pump the positioning of the syringe tip (horizontal or vertical) made no difference in fat concentration at an infusion rate of 1 ml/hr, whereas at 4 and 7 ml/hr the fat concentration was increased significantly by keeping the syringe tip vertical. With either mechanical pump a large fat bolus was delivered during the eighth and final hour of infusion if the milk remaining in the tubing was recovered by using air infusion at the same infusion rate. Intermittent bolus delivery of human milk resulted in no significant loss of human milk fat, no changes in fat concentration, and no terminal delivery of a large fat load. Thus intermittent bolus feedings are preferred over continuous mechanical pump infusion systems for the delivery of human milk to low-birth-weight infants.

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The ideal postnatal rate of bone mineralization in preterm infants is unknown. In this study the effect of a new formula on bone mineralization in very low-birth-weight infants (28 to 32 weeks' gestation) was studied using infant-adapted direct photon absorptiometry. The 81 calories/dl experimental formula (Similac Special Care) contained 50% of the carbohydrate as Polycose, 50% of the fat as medium-chain triglycerides, and a whey:casein protein ratio of 60:40. It also contained large amounts of calcium (140 mg/dl), phosphorus (75 mg/dl), and vitamin D (120 IU/dl). Over a three- to five-week period, bone mineralization increased in two-thirds of the infants and decreased in the remainder, but all values remained in the range expected for bone mineral content in utero. Serum calcium, phosphate, magnesium, alkaline phosphatase, 25-hydroxy-vitamin D, and parathyroid hormone concentrations remained stable, whereas calcitonin concentrations fell.

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Elevated 1,25 dihydroxyvitamin D concentrations were found in five VLBW infants who developed rickets at two to three months postnatal age or term postconceptual age; 25 hydroxyvitamin D concentrations were low. Bone mineralization was found to be extremely low as measured by infant-adapted direct photon absorptiometry. After treatment with a formula supplemented with additional Ca and P, there was a rapid improvement in bone mineralization with a concomitant decrease of 1,25(OH)2D to normal adult values, whereas 250HD values increased and parathyroid hormone values decreased. In the VLBW infants studied, we suggest that rickets may be caused by Ca and P deficiency rather than by a deficiency of vitamin D metabolism.

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Eighteen term, healthy, appropriate for gestational age, breast-fed infants were studied in a double-blind prospective study to determine whether or not supplemental vitamin D affected bone mineralization. All patients were from a single, private pediatric practice. Nine infants were randomly assigned to a vitamin D supplement of 400 IU/day and nine infants to a placebo. By 12 weeks of age, infants receiving placebo had a significant decrease in bone mineralization and in serum 25-hydroxyvitamin D concentrations compared to the vitamin D-supplemented group. It is not known whether or not the increased BMC at 12 weeks of age in vitamin D-supplemented breast-fed infants is of ultimate value. Supplemental vitamin D may be necessary for optimal bone mineralization in term breast-fed infants. A longer follow-up study and additional analyses are required to make conclusive statements.

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