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.     

Effect of dietary methionine and lysine on the toxicity of ingested lead acetate in the chick

The interaction of methionine or lysine with lead in the diet of chicks was studied. In experiment 1, lead acetate to supply 0 or 1000 ppm lead was added to a diet that was either deficient in the total sulfur-containing amino acid (TSAA) content (62% of the requirement) or supplemented with DL-methionine to provide 100% of the requirement (NRC, 1977). Supplementing the 0 ppm lead diet with methionine improved body weight gain. Dietary addition of 1000 ppm lead significantly decreased body weight gain; however, supplemental methionine partially alleviated the lead-induced growth depression (methionine X lead interaction was significant). Liver glutathione levels were markedly increased by supplemental methionine and also by lead but no methionine X lead interaction was detected. In experiment 2, the interaction between an essential non-sulfur-containing amino acid, lysine, and dietary lead was investigated. Two levels of lead, 0 or 1000 ppm, were fed in diets either deficient or adequate in lysine (85 or 100% of the requirement, respectively). Addition of lysine to the lysine-deficient basal diet increased growth. The magnitude of the lead-induced growth depression was not affected by dietary lysine content. The lysine level of the diet did not influence the liver glutathione concentration.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

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.     

Iron deficiency and marginal vitamin A deficiency affect growth,hematological indices and the regulation of iron metabolism genes in rats

Iron deficiency and marginal vitamin A (VA) deficiency frequently coexist and affect billions of people, mostly children and women, worldwide. The effects of these micronutrient deficiencies alone and in combination on hematologic, biochemical and molecular indices of iron and VA status were investigated in a 2 x 2 randomized blocked study conducted in growing male Sprague-Dawley rats. From 3-8 wk of age, rats were fed one of four purified diets that were either adequate or restricted in iron (Fe) and adequate or marginal in VA: (+)Fe(+)VA, 20.3 and 0.367 micro g/g, respectively, denoted control diet; (-)Fe(+)VA, 3.34 and 0.405 micro g/g; (+)FeVA(m), 22.2 and 0.051 micro g/g; or (-)FeVA(m), 3.03 and 0.055 micro g/g. Weight-matched rats fed adequate micronutrients were included to control for possible confounding effects of Fe deficiency on growth and feed efficiency. Iron restriction reduced (P < 0.05) weight gain, feed efficiency, blood hemoglobin and hematocrit. Plasma and liver iron and plasma transferrin saturation were reduced by approximately 50%, whereas liver transferrin mRNA and protein and transferrin receptor mRNA were elevated, as was liver ferritin light-chain protein and light-chain mRNA. Liver heavy-chain ferritin mRNA, hemopexin, ceruloplasmin and cellular retinol-binding protein mRNA were not affected by iron or VA restriction. Although marginal VA deficiency did not exacerbate indices of poor iron status during iron deficiency, iron deficiency was associated with lower plasma retinol and elevated liver VA concentrations. These results are consistent with an impaired mobilization of liver retinol during iron deficiency as well as multiple alterations in iron metabolism.     

Serum and secretory proteins in iron-deficient rat pups and dams

The effects of iron status during reproduction on serum and secretory proteins in rat dams and pups were studied. Pregnant rats were fed ad libitum diets containing 10 or 250 ppm iron throughout gestation and lactation. Litters were adjusted on day 1 to contain 7 pups, and on day 17, dams and pups were sacrificed. Iron status was determined, and concentrations of various serum and secretory proteins were measured. Iron-deficient pups had lower hemoglobin, serum iron, and liver iron compared to controls. Serum albumin and globulin concentrations were significantly increased in iron-deficient pups. Pup serum lysozyme and myeloperoxidase levels were unchanged by iron deficiency. The serum protein profile of dams was relatively unaltered by dietary iron deficiency. Milk iron concentration was significantly decreased in iron-deficient dams; however, milk from all dams was similar in concentration of three immunoproteins measured: lysozyme, peroxidase, secretory IgA. Salivary total protein, lysozyme, and secretory IgA concentrations were similar between groups of dams. It is concluded that dietary iron deficiency during reproduction which does not retard growth of pups but does deplete iron stores has a minimal effect on the secretory immunoproteins measured.     

The effect of dietary iron levels on changes in iron status and zinc-dependent enzyme activities in rats fed two levels of dietary zinc

In order to determine the interrelationship between dietary iron and zinc levels, the effects of dietary iron levels (2, 10, 20, and 40 microg/g) on changes in iron and zinc status and zinc enzyme activities (aminolevulinic acid dehydratase ALA-D EC 4.2.1.24 and alkaline phosphatase ALK-P EC 3.1.3.1) in male Wistar rats were investigated using adequate and marginally deficient zinc diets (25 and 5 microg/g). When rats were fed 5 microg Zn/g diets, body weight gain and food intake remained unchanged at a Fe diet intake of 20 microg/g or greater. Similar tendencies were obtained for hemoglobin, hematocrit, plasma iron, and transferrin saturation. In contrast, liver, spleen, and femur iron concentrations increased gradually with increased iron intake. Feeding diets containing 25 microg Zn/g did not alter these parameters. The percentages of apparent iron absorption in both dietary zinc groups tended to increase with decreasing dietary iron and attained maximum levels at an Fe intake of 10 microg/g. However, In the case of rats fed Fe at concentrations of 2 microg/g Iron absorption decreased. Regardless of the dietary zinc level, rats fed diets with an Fe concentration of 2 microg/g had decreased zinc absorption and plasma ALK-P activity. However, ALA-D activity was not influenced by dietary iron.     

Influence of maternal magnesium deficiency on tissue lead content of rats

Diets containing 150 or 600 ppm magnesium with or without 200 ppm lead were fed to rats throughout gestation and lactation to determine the influence of moderate magnesium deficiency on tissue lead content of maternal and offspring tissue. During lactation it was necessary to increase the lowest dietary magnesium level to 225 ppm. Lead caused a significant depression in both gestational weight gain and average pup weight regardless of the level of dietary magnesium. Maternal magnesium deficiency was evidenced by significant reductions in serum and tibia magnesium, a 17-fold increase in kidney calcium, and hyperemia of the ears. In offspring, however, only growth and tibia magnesium were significantly affected by the magnesium deficiency, and the maternal-fetal difference in serum and tibia magnesium concentration was maintained. Maternal magnesium deficiency resulted in significantly higher lead concentrations in dam liver, and offspring erythrocytes, liver and tibia. A mechanism for the enhanced accumulation of lead in maternal and offspring tissue as a result of maternal magnesium deficiency is not defined, but it is likely to involve enhanced intestinal lead absorption.     

Level of dietary iron,not type of dietary fat,is hyperlipidemic in copper-deficient rats

OBJECTIVE: This study was conducted to determine whether high dietary iron will negate the protective effect of unsaturated fat against hyperlipidemia. METHODS: Forty-eight weanling, male Sprague Dawley rats were randomly assigned to eight dietary groups differing in the levels of copper and iron and type of dietary fat (saturated or unsaturated). The diets were either deficient (0.6 microg Cu/g) or adequate (6.8 microg Cu/g) copper and either adequate (53 microg Fe/g) or high (506 microg Fe/g) iron. All diets contained starch as the sole source of dietary carbohydrate. RESULTS: Regardless of the type of dietary fat, three copper-deficient rats fed the high levels of dietary iron died prematurely due to ruptured hearts. Surviving rats belonging to the copper deficiency and high-dietary iron regimen developed severe anemia, enlarged hearts and livers, and exhibited the highest levels of liver iron. These rats also developed hypercholesterolemia. Triglycerides were elevated by the consumption of high iron diets. CONCLUSION: Data show that levels of dietary iron, not the type of dietary fat, are potential inducers of hypertriglyceridemia. Data also show that the combination of high iron intake and dietary copper deficiency is responsible for elevating blood cholesterol.     

铁、铜对过量锌饲养小鸡生物效应的影响

本文对饲料中过量锌对小鸡铁,铜利用的影响以及补充适量铜或铁、铜的预防效果进行了研究。一日龄雄性小鸡喂基础饲料5天后按体重分成4组,用铁锌铜含量不同的实验饲料喂养28天,观察小鸡生长情况及血液生化指标。结果表明,高锌组小鸡生长不良,Hb下降13％,RBC,Hct,MCV均降低、FEP增加,呈现典型小细胞低色素贫血态,血液及肝肾中锌含量增加,铁和铜含量减少;红细胞超氧化物岐化酶和血浆铜兰蛋白活性下降。在含锌饲料中补充铜能改善小鸡的生长状况,体内铜水平及铜依赖酶活性,对改善贫血及体内铁水平的作用不明显。同时补铁则小鸡各项实验指标均得到明显改善。     

Absorption and metabolic efficiency of iron in nickel deficiency]

Absorption and Metabolic Efficiency of Iron During Ni Deficiency. Ni deficiency leads to reduced iron contents in organs and to greatly reduced Hb levels and erythrocyte counts. Using models it was studied whether this Ni-dependent Fe anemia can be attributed to an impaired absorption of iron or to its metabolic efficiency. An experiment with seven 30-day-old rats from each of two generations were used for this. In Ni deficiency (0.015 ppm dietary nickel) iron absorption was clearly impaired at both 50 ppm and 100 ppm iron in the diet. Compared to the groups given 20 ppm nickel, the amount of iron absorbed fell two-thirds and one-third, respectively. By comparison, the influence on the metabolic efficiency of the iron was relatively small; at high iron supply, however, it was reduced by 8% in the Ni-deficient animals. Therefore, the reduced levels of hemoglobin, erythrocytes and hematocrit must essentially be caused by the impared absorption.     

Level of Dietary Iron,Not Type of Dietary Fat,is Hyperlipidemic in Copper-Deficient Rats

Objective: This study was conducted to determine whether high dietary iron will negate the protective effect of unsaturated fat against hyperlipidemia.

Methods: Forty-eight weanling, male Sprague Dawley rats were randomly assigned to eight dietary groups differing in the levels of copper and iron and type of dietary fat (saturated or unsaturated). The diets were either deficient (0.6 μg Cu/g) or adequate (6.8 μg Cu/g) copper and either adequate (53 μg Fe/g) or high (506 μg Fe/g) iron. All diets contained starch as the sole source of dietary carbohydrate.

Results: Regardless of the type of dietary fat, three copper-deficient rats fed the high levels of dietary iron died prematurely due to ruptured hearts. Surviving rats belonging to the copper deficiency and high-dietary iron regimen developed severe anemia, enlarged hearts and livers, and exhibited the highest levels of liver iron. These rats also developed hypercholesterolemia. Triglycerides were elevated by the consumption of high iron diets.

Conclusion: Data show that levels of dietary iron, not the type of dietary fat, are potential inducers of hypertriglyceridemia. Data also show that the combination of high iron intake and dietary copper deficiency is responsible for elevating blood cholesterol.     

Prolonged acetaminophen ingestion by mice fed a methionine-limited diet does not affect iron-induced liver lipid peroxidation or S-adenosylmethionine.

This study determined whether acetaminophen (ACAP)-induced glutathione depletion was associated with liver lipid peroxide formation, or the concentrations of liver S-adenosylmethionine and S-adenosylhomocysteine in mice fed diets with L-methionine below or at the requirement level (0.25 or 0.5%) for 7 wk. Iron dextran (281 mg/kg body wt) or saline was administered for 2 d before measurement of lipid peroxide formation. Chronic dietary ACAP (0.5%) in mice fed 0.25% methionine caused a failure to maintain body weight even though food intake was similar to intake by all other treatment groups. Liver GSH (measured as nonprotein sulfhydryl concentration) and cysteine concentrations were depleted by ACAP and by ACAP plus iron. Liver lipid peroxide formation was increased by iron but was not altered additionally by ACAP ingestion. Liver glutathione peroxidase activity was increased by methionine in controls, whereas glutathione S-transferase activity was increased by ACAP ingestion in mice fed 0.5% methionine compared with controls. Liver S-adenosylmethionine and nuclear 5-methyldeoxycytidine concentrations were not affected by dietary ACAP or methionine. Liver S-adenosylhomocysteine levels were lower in mice fed ACAP and 0.25% methionine compared with mice fed ACAP and 0.5% methionine. In conclusion, chronic ACAP did not increase the susceptibility of mice to liver lipid peroxidation or alter the availability of methyl groups for methylation reactions.     

Postweaning carnitine supplementation of iron-deficient rats

Severe iron deficiency in the suckling and weanling rat is associated with lipid accumulation in serum and liver, impaired ketogenesis in the suckling pup and low levels of carnitine in some tissues. Carnitine has been effective in reducing high triacylglycerol levels in humans and rats. This study examined tissue triacylglycerol concentrations of iron-deficient rats supplemented with carnitine or iron. Iron-adequate (C) and iron-deficient (D) pups were weaned to diets containing 38 ppm Fe (c) or 6 ppm Fe (d) with or without 0.2% DL-carnitine (Carn) resulting in six experimental treatments: CcCarn, DdCarn, Cc, Cd, Dc, Dd. Males received the diets for 2 wk and female littermates for 4. After 2 and 4 wk, carnitine supplementation significantly increased carnitine content in liver, heart and skeletal muscle by 30-60% in rats from control and Fe-deficient dams. Carnitine treatment significantly lowered the triacylglycerol level in liver of 49-d-old Fe-deficient females, but did not affect other tissues at either time point compared to other dietary treatments. Fe supplementation did not increase carnitine content in tissues, but did reduce triacylglycerol levels in liver by 4 wk and in skeletal muscle at both time points. Possible mechanisms by which iron and carnitine may lower lipids are discussed.     

Effects of developmental iron deficiency and post-weaning iron repletion on the levels of iron transporter proteins in rats

BACKGROUND/OBJECTIVESIron deficiency in early life is associated with developmental problems, which may persist until later in life. The question of whether iron repletion after developmental iron deficiency could restore iron homeostasis is not well characterized. In the present study, we investigated the changes of iron transporters after iron depletion during the gestational-neonatal period and iron repletion during the post-weaning period.MATERIALS/METHODSPregnant rats were provided iron-deficient (< 6 ppm Fe) or control (36 ppm Fe) diets from gestational day 2. At weaning, pups from iron-deficient dams were fed either iron-deficient (ID group) or control (IDR group) diets for 4 week. Pups from control dams were continued to be fed with the control diet throughout the study period (CON).RESULTSCompared to the CON, ID rats had significantly lower hemoglobin and hematocrits in the blood and significantly lower tissue iron in the liver and spleen. Hepatic hepcidin and BMP6 mRNA levels were also strongly down-regulated in the ID group. Developmental iron deficiency significantly increased iron transporters divalent metal transporter 1 (DMT1) and ferroportin (FPN) in the duodenum, but decreased DMT1 in the liver. Dietary iron repletion restored the levels of hemoglobin and hematocrit to a normal range, but the tissue iron levels and hepatic hepcidin mRNA levels were significantly lower than those in the CON group. Both FPN and DMT1 protein levels in the liver and in the duodenum were not different between the IDR and the CON. By contrast, DMT1 in the spleen was significantly lower in the IDR, compared to the CON. The splenic FPN was also decreased in the IDR more than in the CON, although the difference did not reach statistical significance.CONCLUSIONSOur findings demonstrate that iron transporter proteins in the duodenum, liver and spleen are differentially regulated during developmental iron deficiency. Also, post-weaning iron repletion efficiently restores iron transporters in the duodenum and the liver but not in the spleen, which suggests that early-life iron deficiency may cause long term abnormalities in iron recycling from the spleen.     

Influence of dietary lipids on iron and copper levels of rats administered oral contraceptives.

Interrelationships between oral contraceptives and dietary lipids on iron and copper levels in plasma and tissues were investigated in rats. Diets containing either 20% (by weight) safflower oil or hydrogenated coconut oil with and without cholesterol (0.5%) were fed to weanling, female, Wistar-strain rats for a period of 19 weeks. Three types of oral contraceptive agents differing in estrogen/progesterone ratios were administered during weeks 16 through 19 of the experiment. Control rats received the dietary treatment without oral contraceptives. Hemoglobin concentration, hematocrit, red blood cell counts, mean cell hemoglobin and hemoglobin concentration, and mean cell volume values were similar among the various dietary and drug-treatment groups. Elevated levels of copper were found in livers of drug-treated animals fed diets containing cholesterol and safflower oil, whereas levels of copper or iron in spleen and kidney were not influenced by oral contraceptives. Dietary safflower or coconut oil had no influence on levels of iron or copper in plasma. However, iron levels were higher in liver, spleen, and kidneys of rats fed coconut oil compared with those fed safflower oil. Cholesterol-fed rats had reduced levels of iron in plasma and tissues and increased levels of copper in plasma and liver. Iron deficiency in cholesterol-fed rats was indicated by low levels of iron in plasma, liver, spleen, and kidney. In experiment 2, animals were fed the 20% safflower oil diet, with and without sodium glycocholate or cholesterol, to determine whether the apparent malabsorption of iron resulted from sodium glycocholate or cholesterol. Sodium glycocholate resulted in a marked increase in the absorption of iron, whereas cholesterol depressed absorption.     

Influence of iron and the sex of rats on hematological, biochemical and immunological changes during copper deficiency

The influence of dietary iron and the sex of rats on suppressed lymphocyte functions caused by copper deficiency was examined. Male and female weanling Lewis rats were fed two concentrations of copper (0.6 or 5.6 micrograms Cu/g diet) and iron (50 or 300 micrograms Fe/g diet) for 42 d. Regardless of dietary iron concentrations, male and female rats consuming low copper diets had lower serum ceruloplasmin activity and serum and liver copper concentrations than those fed the high copper diet. However, hemoglobin and hematocrit levels were higher in copper-deficient females than in copper-deficient males and were unaffected by copper deficiency in females fed the high iron diet. Copper-deficient females also had higher serum and liver iron concentrations than copper-deficient males. Proliferation of concanavalin A (Con A)-stimulated spleen lymphoid cells (SLC) was suppressed in copper-deficient males and females, but the suppression was less in the females. Thus, the primary cause of suppressed SLC proliferation in copper-deficient rats is poor copper status; poor iron status induced by copper deficiency had little influence on proliferation.     

Cellular growth in iron-deficient rats: effect of pre- and postweaning iron repletion

Effects of iron deficiency and repletion pre- and postweaning on cell growth in young rats were studied. Pregnant dams were fed 6 or 250 ppm iron. On d 2 of lactation, half of the dams in each group were fed the opposite diet. On d 17, cell growth in the crossed-over groups was similar to controls showing that cellular development is impaired only when the iron deficiency is present during gestation and lactation. In a second experiment pup littermates of dams fed 6 (D), 12 (M) and 250 (C) ppm iron were weaned to either the same diet as fed to their dams DD, MM or CC; repleted with iron DC, MC; or fed the deficient diet CD until 42 d of age. After dietary iron repletion, cell numbers in thymus (DC and MC) and liver (DC) were greater than those of deficient littermates, but were less than those of controls (CC). Iron repletion postweaning reduced the cardiac hypertrophy (DC vs. DD and MC vs. MM) and increased splenic cell number compared to unrepleted deficient littermates (DC vs. DD). Thus, the severity and reversibility of impaired cellular growth is dependent on the timing and severity of the deficiency and the organ affected.     

Feeding low iron diets affects fluid preference in growing rats

The effect of dietary iron (ferrous sulfate) concentration on the intake of solutions of sodium saccharin, hydrochloric acid, quinine sulfate, sodium chloride, and potassium chloride was studied. Male Charles River CD weanling rats were divided into four groups and fed semi-synthetic iron-free diets supplemented with either 0, 10, 20 or 40 ppm ferrous sulfate. Using a 48 hour, two-bottle preference test, the rats fed the diet with 0 ppm (added) iron showed significantly higher preferences for sodium chloride and potassium chloride solutions compared with the other three groups. Total fluid intake also increased in rats given the unsupplemented diet. Water intake was not increased in this 0 ppm group when sodium was offered. Blood hemoglobin and hematocrit, serum iron and body weight were lowest in the group with 0 ppm added iron, yet neither adrenal weights nor serum zinc was affected by the dietary iron levels.     

Low copper and brain abnormalities in fetus from triethylene tetramine dihydrochloride-treated pregnant mouse.

The effects of prenatal triethylene tetramine dihydrochloride (Trien-2HCl) exposure on fetal mice have been investigated on gestational day 19. Trien-2HCl was given throughout pregnancy at levels of 0 (control), 3,000, 6,000, or 12,000 ppm as drinking water, ad libitum. At the level of 12,000 ppm, the frequency of total resorption tended to be high and that of fetal viability tended to be low, as compared to controls. Decreased maternal weight was observed in body, but not in liver, at the level of 12,000 ppm. Fetal body and cerebrum weights significantly decreased at the levels of 6,000 and 12,000 ppm; however, fetal liver weight remained unchanged. Maternal serum copper concentration was not affected by the Trien-2HCl. Fetal copper concentrations of liver and cerebrum were significantly lower in the Trien-2HCl-treated groups than in the controls, with levels decreasing in a dose-related manner. When the copper and zinc concentrations in the group treated at 12,000 ppm were compared with those in controls, significant decreases in both metals were observed in placenta but not in maternal liver. Changes in fetal zinc concentration varied by tissues: i.e., an increase in liver and no change in cerebrum. Fetal abnormalities were frequently observed in brain, and the frequency was increased with increasing levels of the Trien-2HCl. These results suggest that fetal brain abnormalities caused by Trien-2HCl may be due in part to induction of copper deficiency, which is almost equivalent to that in brindled mutant mouse.