Development of anemia in copper-deficient rats fed high levels of dietary iron and sucrose

The effects of dietary carbohydrate and iron on the development of copper deficiency were examined. Male Sprague-Dawley rats (n = 48) were limit-fed one of eight diets in a 2 X 2 X 2 factorial design for 19 d. Two levels of copper (0.85 or 8.6 micrograms Cu/g diet) and iron (54 or 226 micrograms Fe/g diet) and two types of carbohydrate (sucrose or cornstarch, 65.3%) were fed. Compared with control rats, copper-deficient rats had lower hematocrits, lower ceruloplasmin levels, lower tissue levels of copper and increased hepatic iron levels. Copper-deficient rats fed sucrose had significantly lower hematocrits, lower apparent absorption of copper, lower liver iron levels and higher plasma triglyceride levels than copper-deficient rats fed cornstarch. Copper-deficient rats fed sucrose with 226 micrograms Fe/g diet had hematocrit levels that were 15% lower than all other copper-deficient levels and 23% lower than control levels. Tissue levels of copper among copper-deficient rats were not affected by the type of carbohydrate or by the level of dietary iron. These data indicate that both high iron and sucrose can affect the development of the copper deficiency.     

Determinants of copper-deficiency anemia in rats

Indicators of copper and iron metabolism were studied in pregnant rats and their 90-d-old offspring fed copper-sufficient or copper-deficient diets containing marginal or adequate levels of iron from the beginning of pregnancy until the offspring were 90 d of age. Offspring had more severe signs of copper deficiency (including anemia, hypertrophy of the heart, decreased activity of ferroxidase I and II, depression of growth and death) than the dams. In both dams and offspring, copper deficiency resulted in anemia when dietary iron was marginal but not when it was adequate. Liver iron was elevated in copper-deficient male offspring, but not in female offspring. Anemia and growth retardation were more pronounced in copper-deficient males than in females, despite similarly low levels of ferroxidase I and II. Iron absorption was reduced by copper deficiency only in female offspring. Activity of 59Fe in various tissues 6 or 48 h after gavage did not reveal any other effect of copper deficiency on iron metabolism. Thus age at the time copper-deficient diets were introduced, sex and dietary iron strongly influence the effect of copper deficiency.     

Lipid alterations in the liver and serum of rats in histidine-excess and copper deficiency

To obtain further information on lipid metabolism in the histidine-excess and copper-deficiency, rats were fed basal, histidine-excess (the addition of 50 g L-histidine/kg diet) or copper-deficient diets for 0, 7, 21 and 42 d ad libitum. Liver triacylglycerol accumulated and the serum triacylglycerol level decreased after feeding of the histidine-excess diet for 21 or 42 d, but not after feeding of the copper-deficient diet. Serum cholesterol level increased in rats fed the histidine-excess diet for 7, 21 and 42 d, but not in rats fed the copper-deficient diet. Copper content in the liver and serum significantly decreased in rats fed the histidine-excess diet. Copper content in the liver and serum was markedly decreased in rats fed the copper-deficient diet. Liver zinc content was constant, but the serum zinc level decreased in rats fed the histidine-excess diet. Feeding of the copper-deficient diet hardly affected zinc content in the liver and serum. Urinary copper and zinc increased in rats fed the histidine-excess diet, and decreased or showed a decreasing tendency in rats fed the copper-deficient diet. Overall results indicated that feeding the histidine-excess diet caused copper deficiency, whereas hypercholesterolemia was not shown in rats fed the copper-deficient diet although the livers of rats fed the copper-deficient diet contained less copper than those of rats fed the histidine-excess diet. Thus, the responses on liver triacylglycerol and serum cholesterol to copper deficiency induced by the feeding of a histidine-excess diet are different from those to copper deficiency induced by feeding of a copper-deficient diet.     

Oxidation, excretion, and tissue distribution of [26-14C] cholesterol in copper-deficient rats

The effect of copper deficiency on in vivo catabolism and excretion of [26-14C] cholesterol was studied in male rats. The study involved four treatments, namely, control, copper-deficient, control plus cholesterol, and copper-deficient plus cholesterol supplement. Significant elevations of serum ester and total cholesterol concentrations and reductions of serum free, ester, and total cholesterol specific activities were observed in rats fed the copper-deficient diets. In addition, a significant reduction of liver free cholesterol concentration was observed in rats fed the copper-deficient diets. Cholesterol supplementation significantly increased the concentrations of, and reduced the specific activities of, the different serum and liver cholesterol fractions. The only exception was that the liver free cholesterol concentration was not altered by cholesterol supplementation. The serum free cholesterol concentration was significantly increased and the specific activities of liver ester cholesterol were significantly reduced in rats fed the copper-deficient diet with no added cholesterol. The rates of oxidation and excretion of [26-14C] cholesterol were not influenced by dietary copper but were significantly increased by cholesterol supplementation. A shift of cholesterol from the liver to the serum pool appeared to be responsible for the hypercholesterolemia observed in copper deficiency.     

Modification of vitamin A metabolism in rats fed a copper-deficient diet

The liver is the main storage site of vitamin A and copper. Inverse relationships between copper and vitamin A liver concentrations have been suggested. We have investigated the consequences of a copper-deficient diet on liver and blood vitamin A storage in Wistar rats. Animals were fed either a copper-deficient diet for 45 days from weaning, or an identical diet containing adequate amounts of copper. Concentrations of vitamin A were determined by isocratic high performance liquid chromatography using UV detection. We have observed in the liver of the rats fed a copper-deficient diet a significantly higher mean level of retinyl esters (148 +/- 37 micrograms/g of liver) and retinol (3.3 +/- 1.4 micrograms/g of liver) compared to the mean concentration of the retinyl esters (53 +/- 8.5 micrograms/g of liver) (p less than 0.01) and retinol (1.4 +/- 0.5 micrograms/g of liver) (p less than 0.01) in controls. Opposite results were observed in the serum of the group fed a copper-deficient diet as these rats had a significantly lower level of retinol (22 +/- 4 micrograms/100 ml) compared to the mean concentration in the controls (64 +/- 20 micrograms/100 ml) (p less than 0.01). These findings suggest that a copper-deficient diet may cause defective transport of vitamin A from liver to blood. This experimental model may be useful to further investigate unusual liver vitamin A and copper concentrations observed in children during various hepatobiliary diseases.     

Copper and iron deficiencies: effects on serum lipids and tissue minerals

To study the interrelationship between iron and copper on serum lipid concentrations, four diets were fed to growing rats: iron and copper deficient, copper-deficient, iron-deficient, iron and copper adequate. After 18 weeks, concentrations of iron and copper in organs and lipids in sera were determined. Iron deficiency alone or combined with copper deficiency resulted in reduced body weights, hemoglobin concentrations, hematocrits, and iron concentrations in liver, spleen, and heart. Hepatic copper was elevated 8-fold in iron deficiency. Copper deficiency alone or combined with iron deficiency resulted in reduced copper concentrations of liver and spleen and reduced ceruloplasmin. Serum triglycerides and cholesterol did not differ among experimental treatments. No significant effects of the interaction between dietary iron and copper on serum lipid levels were found.     

Interaction of dietary carbohydrate, ascorbic acid and copper with the development of copper deficiency in rats

The effects of dietary carbohydrate and ascorbic acid on the development of copper deficiency were investigated. Male Sprague-Dawley rats (n = 48) were fed one of eight diets in a 2 X 2 X 2 factorial design for 21 d. These diets varied in copper (1.11 or 8.96 micrograms Cu/g diet), carbohydrate (sucrose or cornstarch, 62.3%) and ascorbic acid (0 or 1%). Compared to controls, copper-deficient rats had lower hematocrit and ceruloplasmin levels, lower levels of copper and iron in several tissues, higher heart weights and lower spleen weights. During copper deficiency, liver iron levels were higher than control levels when cornstarch, but not sucrose, was the carbohydrate source, while liver and gastrointestinal tract weights were higher with sucrose compared to cornstarch. Copper-deficient rats fed ascorbic acid had significantly (P less than 0.05) lower hematocrits when fed sucrose compared to starch [29.6 +/- 1.2 vs. 36.8 +/- 1.2 g/dl (mean +/- SEM), respectively]. In copper-deficient rats, sucrose tended to lower the apparent absorption of copper compared to cornstarch, while ascorbic acid reduced the apparent absorption of iron. Thus, sucrose and ascorbic acid appeared to reduce hematocrit levels through effects on mineral absorption.     

Influence of sucrose and starch on the development of anemia in copper- and iron-deficient rats

The purpose of this investigation was to compare the effects of sucrose and starch on the development of copper and iron deficiency. Male Sprague-Dawley rats (n = 48) were fed one of eight diets in a 2 X 2 X 2 factorial design for 24 d. Two levels of copper (deficient, 0.7 microgram/g, or adequate, 8.3 micrograms/g) and iron (deficient, 8.3 micrograms/g, or adequate, 50 micrograms/g), and two types of carbohydrate (sucrose or starch, 62% of the diet) were fed. Copper-deficient rats had significantly lower hematocrit, hemoglobin and tibia iron levels and depressed copper and iron absorption when fed sucrose instead of starch. The apparent absorption of copper, but not iron, was significantly lower when rats deficient in both copper and iron were fed sucrose rather than starch. Iron-deficient rats fed sucrose apparently absorbed significantly more iron than those fed starch; however, sucrose did not significantly improve hematocrit and hemoglobin levels. The metabolism of copper and iron by rats fed diets adequate in these nutrients was not affected by the type of dietary carbohydrate. These data indicate that the type of dietary carbohydrate alters both copper and iron metabolism, particularly in copper-deficient rats.     

Female rats are protected against oxidative stress during copper deficiency

BACKGROUND: Copper deficiency induces a dramatic decrease of superoxide dismutase activity and leads to alteration of antioxidant defense systems. METHODS: and Objective: Experiments were conducted in weanling male, intact and ovariectomized female rats, fed either a copper-adequate or copper-deficient diet for seven weeks, in order to determine whether endogenous estrogen could modulate oxidative stress and the severity of copper-deficiency. RESULTS: Feeding male rats a copper-deficient diet induced typical signs of copper deficiency, such as decreased hepatic copper, growth retardation, anemia, heart hypertrophy, pancreas atrophy and hypercholesterolemia. Furthermore, copper deficiency increased the amount of lipid peroxidation products in the heart, liver and pancreas following in vitro iron induction. Although levels of hepatic copper in copper-deficient females were similar to those of their male counterparts, the females were partially protected from the adverse effects of the deficiency (no growth retardation, less severe anemia, lesser extent of lipid peroxidation). Thus, female rats are provided with a greater degree of protection against oxidative damage than males. However, females did not appear to be protected against pancreas atrophy, heart enlargement and hypercholesterolemia induced by copper deficiency. This observed partial protection of females was lost after ovariectomy as shown by decreased body weight and hematocrit, heart enlargement and higher tissue peroxidation in ovariectomized females compared to intact females. CONCLUSIONS: The results suggest that the partial protection of copper deficient females is related to the antioxidant properties of estrogens. The protective action of estrogen against oxidative stress is of particular importance when antioxidant defenses are decreased as shown in this experimental model.     

Effects of varying dietary iron on the expression of copper deficiency in the growing rat: anemia, ferroxidase I and II, tissue trace elements, ascorbic acid, and xanthine dehydrogenase

The effect of dietary iron on the development of copper-deficiency anemia in the growing rat was investigated. For up to 80 d, female rats (75 g) were fed purified diets containing adequate, marginal or low levels of iron, and either 0.7 or 10 ppm copper. Hemoglobin levels and factors postulated to affect liver iron mobilization, including ferroxidase (Fox) I and II, ascorbate and liver xanthine dehydrogenase (XDH) were assayed. By d 7, Fox I activity in the copper-deficient groups was 10% that of the copper-sufficient groups; thereafter, Fox I activity remained low, and was not affected by dietary iron. Fox II activity in the copper-deficient groups after d 28 was 50-75% of values from rats adequate in copper. On d 49, hemoglobin levels in the copper-deficient groups were lower than in the copper-sufficient groups fed low and marginal levels of iron, but were similar to those fed adequate iron. Liver iron was similar in both groups fed adequate iron, but was higher in the copper-deficient than in the copper-sufficient rats fed low or marginal levels of iron. Copper deficiency tended to result in slightly lower ascorbate levels on d 80 at all levels of iron. Liver XDH activity tended to be lower in the copper-deficient groups than in the copper-sufficient groups on d 28 and 49. These results show that copper deficiency may impair liver iron mobilization in the growing rat if dietary iron is low. Possible mechanisms include decreased Fox activity and/or decreased iron reduction by ascorbate or XDH.     

Hepatic cytochrome P-450 and microsomal heme oxygenase in copper-deficient rats

Copper deficient and control rats were pair-fed from weaning a milk-based diet. Each animal received 25 mg of iron parenterally. After 6 weeks of dietary manipulations the animals were killed. As expected, rats fed the copper-deficient diet were anemic. Copper-dependent enzyme activities and copper content of plasma and liver confirmed that copper deficiency was present in these rats. Hepatic microsomal cytochrome P-450 specific contents were similar in control and copper-deficient animals in the uninduced state and following induction of cytochrome P-450 by treatment with phenobarbital. However, microsomal heme oxygenase was increased in copper-deficient animals compared with controls whether they were treated with phenobarbital or not. No differences were observed in splenic microsomal heme oxygenase between dietary groups. These studies suggest that hepatic heme catabolism is enhanced in copper deficiency. The explanation for this enhancement may be related to increased hepatic iron accumulation in copper-deficient animals or to the associated defect of selenium metabolism in copper deficiency.     

Serum cholesterol levels are not elevated in young copper-deficient rats, mice or brindled mice

Experiments were conducted with suckling male C57BL mice and Sprague-Dawley rats to investigate the relationship between copper deficiency and elevated serum cholesterol. Brindled mice, which have a genetic defect that affects copper distribution, were compared to their normal brothers. Dietary copper deficiency was produced in dams heterozygous for the brindled gene, in normal mouse dams and in rat dams. The subsequent male offspring were compared to those from copper-supplemented dams. Copper deficiency, as assessed by liver copper levels or ceruloplasmin activity, was demonstrated in 12-d-old rats, brindled mice, and in genotypically normal mice from dams fed the copper-deficient diet. However, serum cholesterol levels were not elevated in these "copper-deficient" rats or mice. In one experiment serum cholesterol levels of brindled mice were significantly lower than that of their littermate controls. An additional study was done with older mice. Their dams were fed a low copper diet from parturition throughout lactation, and the pups were fed the same copper-deficient diet for 4 wk after weaning. The 7-wk-old male copper-deficient mice had liver copper levels below 1 microgram/g, but no elevation in serum cholesterol was observed. The failure to demonstrate a rise in serum cholesterol in these perinatal models may be due in part to less severe hepatic copper deficiency because of neonatal copper reserves in liver.     

Dietary supplementation with t-butylhydroquinone reduces cardiac hypertrophy and anemia associated with copper deficiency in rats

The effect of dietary supplementation with the antioxidant, t-butylhydroquinone (TBHQ), on some of the cardiovascular consequences of copper deficiency was investigated. Rats were fed copper-deficient (CuD) diet containing 0.3 μg Cu/g of diet that were either nonsupplemented or supplemented with TBHQ (supplied in the dietary safflower oil at a concentration of 0.02%). Control rats were fed copper adequate (CuA) diet containing >5.0 μg Cu/g (CuA) that also were either nonsupplemented or supplemented with TBHQ. After five weeks, rats consuming CuD diet supplemented with TBHQ exihibited plasma copper concentrations, ceruloplasmin activities, and liver and heart copper concentrations that were significantly (P<0.05) lower than those of rats consuming either nonsupplemented or TBHQ supplemented CuA diet, but no different from those of rats consiming nonsupplemented CuD diet. However, rats consuming CuD diet supplemented with TBHQ had significantly (P<0.05) higher growth, hemoglobin concentrations, hematocrits, and red blood cell qistribution widths but lower heart weights than rats consuming nonsupplemented CuD diet. TBHQ supplementation had no effect on these variables in rats fed CuA diet. Thus, while TBHQ did not improve copper status, it did ameliorate the growth reduction, anemia, and cardiac hypertrophy associated with copper deficiency. These findings indirectly support the contention that oxidative damage contributes to the pathophysiological consequences of copper deficiency.     

Effect of copper deficiency on erythrocyte membrane proteins of rats

The effects of copper deficiency on the proteins of rat erythrocyte membranes were assessed by electrophoretic analysis. For 42 d, rats were fed diets containing less than 1 ppm Cu with 35 or 250 ppm Fe or 5 ppm Cu with 35 or 250 ppm Fe. Electrophoresis of erythrocyte membrane proteins indicated a significant increase in the amount of a 170,000-dalton protein in rats fed copper-deficient diets. High dietary iron reduced the amount of the 170,000-dalton protein in erythrocyte membranes from rats in both the copper-deficient and copper-adequate groups. However, feeding high dietary iron to copper-deficient rats did not reduce the amount of the protein to the level found in rats fed the copper-adequate diet containing 35 ppm Fe. Triton X-100 extraction of erythrocyte membranes demonstrated that the 170,000-dalton protein was associated with the membrane cytoskeleton. Thus, copper deficiency possibly alters the cell's mechanical properties and consequently decreases erythrocyte survivability by modifying the membrane cytoskeleton.     

Comparison between dietary and cenetic copper deficiency in mice: Copper-dependent anemia

Copper deficiency was produced by feeding a low copper diet to mice and rats during gestation and lactation. The copper-deficient offspring were compared to Brindled mice, which exhibit copper-deficient characteristics due to a genetic defect within the X-chromosome. Weanling mice nursed by dams fed the low copper diet exhibit classical signs of copper deficiency previously reported in other species, including: anemia, hypoceruloplasminemia, depressed brain copper levels, and cytochrome oxidase activities. However, in comparison to weanling rats derived from dams given the same treatment, the mice show only a minimal growth deficit and marginal copper depletion of liver with normal cytochrome oxidase, activity. Younger 11-day-old copper-deficient male mice also develop anemia, have low tissue copper levels (liver and kidney) and exhibit decreased ceruloplasmin activity. However, unlike copper-deficient rats, these mice do not have a decrease in serum ferroxidase activity. Age-matched Brindled mice are not anemic, even though they have low serum ferroxidase activity. Similar to the dietary deficient mice, Brindled mice do have low ceruloplasmin activity and low liver copper levels. In an attempt to resolve this enigma, mouse serum was chromatographed on Sephadex G-150. Only one ferroxidase activity peak, clearly separated from ceruloplasmin, was observed. Human and rat sera display two ferroxidase activity peaks, one chromatographing with ceruloplasmin activity, which is sensitive to azide, and a second peak which, like the mouse serum peak, appears as a shoulder on the void volume. These results demonstrate the potential of this mouse model in elucidating the nature of copper-dependent anemia. Furthermore, the data from this model suggest that changes in serum ferroxidase activity cannot explain the existence of anemia in dietary copper deficiency nor absence of anemia in genetic copper deficiency (Brindled mice).     

Dietary fructose but not starch is responsible for hyperlipidemia associated with copper deficiency in rats: effect of high-fat diet

OBJECTIVE: To test the hypothesis that copper deficiency in rats may be hyperlipidemic only when the diets consumed contain nutrients which contribute to blood lipids such as fructose and high fat. METHODS: Weanling male Sprague Dawley rats were fed diets which contained either starch or fructose as their sole carbohydrate source. The diets were either inadequate (0.6 microg Cu/g) or adequate (6.0 microg Cu/g) in copper and contained either high (300 g/kg) or low (60 g/kg) fat. At the end of the 4th week the rats were killed. Livers were analyzed for copper content. Plasma was analyzed for cholesterol and triglyceride concentrations. RESULTS: High-fat diet did not increase blood lipids in rats fed a copper-deficient diet containing starch. In contrast, the combination of high-fat diet with fructose increased blood triglycerides and fructose with copper deficiency resulted in a significant increases in blood cholesterol. CONCLUSIONS: Hyperlipidemia of copper deficiency in rats is dependent on synergistic effects between dietary fructose and copper deficiency and fructose and amount of dietary fat. Hyperlipidemia does not develop if starch is the main source of dietary carbohydrate in a copper-deficient diet even if a high-fat diet is fed.     

Total body content of copper and other essential metals in rats fed fructose or starch

The effects of dietary carbohydrate and copper on whole body levels of copper and other essential minerals were investigated. Weaned male rats were pair-fed diets containing 62% fructose or cornstarch with either adequate (6–7 mg/kg) or deficient (0.5–0.7 mg/kg) copper (Cu) for 32–33 days. Recommended levels of other minerals were present in all diets. Total copper in whole body was approximately 2-fold higher in rats fed copper-supplemented diets than copper-deficient diets and was independent of the type of dietary carbohydrate. Whole body levels of zinc, iron, manganese and magnesium were independent of dietary copper and carbohydrate. Selenium content of rats fed copper-deficient diet with fructose was 35% higher than in those fed copper-adequate diet with fructose. Rats fed the copper-deficient diet containing starch had a whole body calcium level 17–23% lower than that of the other dietary groups. These results demonstrate that the copper-carbohydrate interaction is not a result of carbohydrate-dependent effects on the retention of dietary copper and other essential minerals.     

Plasma diamine oxidase activity is greater in copper-adequate than copper-marginal or copper-deficient rats

The object of this study was to determine whether serum diamine oxidase activity could distinguish among adequate, marginal and deficient copper status in rats. Male weanling Sprague-Dawley rats (n = 21) were randomly assigned to one of three dietary regimens, with copper concentrations of 0.52, 1.73 and 6.7 mg/kg diet. On completion of the study, body weights were significantly different among dietary groups, with copper-marginal rats displaying the highest mean weight and copper-deficient rats the lowest. Copper-deficient rats ate significantly less food than the other two groups. Rats fed the three diets had significantly different liver copper concentrations. Liver and heart superoxide dismutase and cytochrome c oxidase activities, and plasma ceruloplasmin and erythrocyte superoxide dismutase activities were significantly lower in the copper-deficient rats than in the other two groups. Plasma diamine oxidase activity was lower in both copper-deficient (0.18 +/- 0.11 U/L) and marginal (0.21 +/- 0.11 U/L) rats compared with copper-adequate rats (3.35 +/- 0.28 U/L). Of the biochemical indices measured, only liver copper concentration (-20%) and plasma diamine oxidase activity (-94%) differed between rats fed copper-marginal and copper-adequate diets. Plasma diamine oxidase activity, therefore, may be a sensitive functional biomarker of suboptimal copper status.     

Tin, copper, iron and calcium metabolism of rats fed various dietary levels of inorganic tin and zinc

Three studies were conducted to determine the effects of various dietary levels of tin (less than 1, approximately 100, approximately 200, approximately 500, approximately 2000 micrograms/g diet) and of zinc (approximately 15, approximately 30, approximately 52 micrograms/g diet) on the metabolism of tin, copper, iron and calcium by growing rats. The accumulation of tin in the kidneys and tibias of animals was proportional to dietary exposure. The concentration of tin in the bones of rats fed greater than 100 micrograms Sn/g diet was 5-fold and 20-fold greater than the levels found in kidney and liver, respectively. Rats fed greater than 500 micrograms Sn/g diet had plasma copper levels that were only 13% of control levels and had depressed copper levels in livers and kidneys. The activity of delta-aminolevulinic acid dehydratase in the erythrocytes of rats fed the highest level of tin was 55% of that found in control animals. The amounts, but not the concentrations, of calcium in the tibias of rats fed greater than 100 micrograms Sn/g diet were less than the levels in the bones of control animals. The moderate variations in dietary zinc levels did not affect significantly the levels of minerals in tissues.     

Zinc-vitamin A interaction in pregnant and fetal rats: supplemental vitamin A does not prevent zinc-deficiency-induced teratogenesis

The influence of a higher-than-normal intake of vitamin A on the detrimental effects of zinc deficiency on vitamin A metabolism was investigated in pregnant Sprague-Dawley rats. At mating, rats were fed diets containing 100 (control), 4.5, or 0.5 micrograms/g zinc combined with 4 (control) or 8 micrograms retinyl acetate/g. Low intake of zinc, but not of vitamin A, caused food intake, total body weight change, fetal weight and placental weight to be low. Incidence of teratogenic effects was more pronounced in low zinc groups than in controls. Concentrations of vitamin A in maternal plasma and liver were affected by the amount of zinc in the diet. Dietary vitamin A, however, did not affect either of these parameters. Maternal plasma zinc concentration was affected only by low dietary zinc, whereas plasma copper and iron were unaffected by the dietary treatments. Maternal liver iron was higher in zinc-deficient rats than in controls; however, maternal liver zinc and copper concentrations were not altered by dietary treatments. No significant differences in vitamin A concentration of fetal liver, fetal plasma, or placenta were seen among the groups. Fetuses from zinc-deficient dams had significantly lower levels of liver vitamin A and liver zinc than did controls. Fetal liver iron was higher in zinc-deficient fetuses than in controls, whereas fetal liver copper was not affected by dietary treatment. These data suggest that supplemental dietary vitamin A does not ameliorate the effect of zinc deficiency on vitamin A metabolism during pregnancy.