Cadmium accumulation and interactions with zinc, copper, and manganese, analysed by ICP-MS in a long-term Caco-2 TC7 cell model

The influence of long-term exposure to cadmium (Cd) on essential minerals was investigated using a Caco-2 TC7 cells and a multi-analytical tool: microwave digestion and inductively coupled plasma mass spectrometry. Intracellular levels, effects on cadmium accumulation, distribution, and reference concentration ranges of the following elements were determined: Na, Mg, Ca, Cr, Fe, Mn, Co, Ni, Cu, Zn, Mo, and Cd. Results showed that Caco-2 TC7 cells incubated long-term with cadmium concentrations ranging from 0 to 10 μmol Cd/l for 5 weeks exhibited a significant increase in cadmium accumulation. Furthermore, this accumulation was more marked in cells exposed long-term to cadmium compared with controls, and that this exposure resulted in a significant accumulation of copper and zinc but not of the other elements measured. Interactions of Cd with three elements: zinc, copper, and manganese were particularly studied. Exposed to 30 μmol/l of the element, manganese showed the highest inhibition and copper the lowest on cadmium intracellular accumulation but Zn, Cu, and Mn behave differently in terms of their mutual competition with Cd. Indeed, increasing cadmium in the culture medium resulted in a gradual and significant increase in the accumulation of zinc. There was a significant decrease in manganese from 5 μmol Cd/l exposure, and no variation was observed with copper.     

Cd, Cu, Zn, Se, and Metallothioneins in Two Amphibians, Necturus maculosus (Amphibia, Caudata) and Bufo bufo (Amphibia, Anura)

The accumulation of cadmium, its affinity for metallothioneins (MTs), and its relation to copper, zinc, and selenium were investigated in the experimental mudpuppy Necturus maculosus and the common toad Bufo bufo captured in nature. Specimens of N. maculosus were exposed to waterborne Cd (85???g/L) for up to 40?days. Exposure resulted in tissue-dependent accumulation of Cd in the order kidney, gills > intestine, liver, brain > pancreas, skin, spleen, and gonads. During the 40-day exposure, concentrations increased close to 1???g/g in kidneys and gills (0.64?C0.95 and 0.52?C0.76; n?=?4), whereas the levels stayed below 0.5 in liver (0.14?C0.29; n?=?4) and other organs. Cd exposure was accompanied by an increase of Zn and Cu in kidneys and Zn in skin, while a decrease of Cu was observed in muscles and skin. Cytosol metallothioneins (MTs) were detected as Cu,Zn?Cthioneins in liver and Zn,Cu?Cthioneins in gills and kidney, with the presence of Se in all cases. After exposure, Cd binding to MTs was clearly observed in cytosol of gills as Zn,Cu,Cd?Cthionein and in pellet extract of kidneys as Zn,Cu,Cd?Cthioneins. The results indicate low Cd storage in liver with almost undetectable Cd in liver MT fractions. In field trapped Bufo bufo (spring and autumn animals), Cd levels were followed in four organs and found to be in the order kidney > liver (0.56?C5.0???g/g >0.03?C0.72???g/g; n?=?11, spring and autumn animals), with no detectable Cd in muscle and skin. At the tissue level, high positive correlations between Cd, Cu, and Se were found in liver (all r?>?0.80; ???=?0.05, n?=?5), and between Cd and Se in kidney (r?=?0.76; n?=?5) of autumn animals, possibly connected with the storage of excess elements in biologically inert forms. In the liver of spring animals, having higher tissue level of Cd than autumn ones, part of the Cd was identified as Cu,Zn,Cd?Cthioneins with traces of Se. As both species are special in having liver Cu levels higher than Zn, the observed highly preferential Cd load in kidney seems reasonable. The relatively low Cd found in liver can be attributed to its excretion through bile and its inability to displace Cu from MTs. The associations of selenium observed with Cd and/or Cu (on the tissue and cell level) point to selenium involvement in the detoxification of excessive cadmium and copper through immobilization.     

Maternal and neonatal scalp hair concentrations of zinc, copper, cadmium, and lead

Postpartum scalp hair samples from 82 term-pregnancy mother/neonate pairs were analyzed for their concentration of zinc (Zn), copper (Cu), cadmium (Cd), and lead (Pb), using inductively coupled plasma-mass spectrometry. Maternal and neonatal Zn concentrations had geometric means (and 99% confidence intervals) of 122.5 μg/g (117.9–131.5 μg/g) and 146.9 μg (141.5–156.7 μg/g) respectively. Corresponding Cu values were 18.4 μg/g (17.6–23.8 μg/g) and 6.7 μg/g (6.3–7.6 μg/g). Those of Cd were 0.49 μg/g (0.47–0.69 μg/g) in the mothers and 0.57 μg/g (0.55–0.86 μg/g) in the neonates. For Pb, they were 7.95 μg/g (7.60–9.32 μg/g) and 4.56 μg/g (4.39–5.56 μg/g). Cigaret smoking, despite its relatively low prevalence (19.5%), was associated with lower Zn and higher Cd and Pb concentrations and in lower Zn/Cd and Zn/Pb molar concentration ratios. Smoking also altered interelemental relationships, particularly those of Zn with Cd and Pb and those between Cd and Pb. Smoking frequency appeared to show negative dose-response effects on maternal and neonatal Zn concentrations, Zn/Pb molar concentration ratios, and birth weight. Mothers with a history of oral contraceptive (OC) usage had significantly higher Cu concentrations and lower Zn/Cu molar concentration ratios than nonusers, with the highest Cu concentrations and lowest Zn/Cu values being associated with third-generation OCs. No similar effects were elicited in the respective neonatal Cu concentrations. Neither alcohol consumption nor prenatal supplementation with iron and/or folic acid had discernible effects on the maternal or neonatal elemental concentrations. The data from this study suggest that in a given population of term-pregnancy mothers and neonates, significant interindividual variations in hair trace element concentrations can occur, irrespective of commonality of general environment, and that lifestyle factors, including cigaret smoking and OC usage history, can be significant contributory factors to such variations. The data are discussed in relation to the effects of smoking-associated exposure to Cd and Pb exposure on Zn availability for placental transfer, as well as on the quantitative maternal Zn supply levels to the fetus resulting from the known tendency of smokers to have lower dietary intakes of Zn. The higher Cu concentrations in OC users are discussed in relation to altered Cu metabolism, characterized by increased synthesis of the Cu-binding protein, ceruloplasmin, as an acute-phase antioxidant response to altered lipid profile and increased lipid oxidation.     

Displacement of zinc and copper from copper-induced metallothionein by cadmium and by mercury: in vivo and ex vivo studies

The in vitro affinity of metals for metallothionein (MT) is Zn less than Cd less than Cu less than Hg. In a previous study Cd(II) and Hg(II) displaced Zn(II) from rat hepatic Zn7-MT in vivo and ex vivo (Day et al., 1984, Chem. Biol. Interact. 50, 159-174). The ability of Cd(II) or Hg(II) to displace Zn(II) and/or Cu(II) from metallothionein in copper-preinduced rat liver (Zn, Cu-MT) was assessed. Cd(II) and Hg(II) can displace zinc from (Zn, Cu)-MT both in vivo and ex vivo. The in vitro displacement of copper from MT by Hg(II) was not confirmed in vivo and ex vivo. Cd(II) treatment did not alter copper levels in (Zn, Cu)-MT, as expected. Hg(II) treatment, however, did not decrease copper levels in MT, but rather increased them. The sum of the copper increase and mercury incorporation into MT matched the zinc decrease under in vivo conditions and actually exceeded the zinc decrease under ex vivo conditions. Short-term exposure of rat liver to exogenous metals can result in incorporation of these metals into MT by displacement of zinc from pre-existing MT. Displacement of copper from pre-existing MT by mercury, as predicted by in vitro experiments, was not confirmed under the conditions of our in vivo and ex vivo experiments. This result is explainable based on the differing affinities and/or preferences of the two metal clusters in MT.     

Oxidative Stress Induced by Cadmium in the C6 Cell Line: Role of Copper and Zinc

In this report, we have investigated the role of copper (Cu) and zinc (Zn) in oxidative stress induced by cadmium (Cd) in C6 cells. Cells were exposed to 20 μM Cd, 500 μM Cu, and 450 μM Zn for 24 h. Then, toxic effects, cellular metals levels, oxidative stress parameters, cell death, as well as DNA damage were evaluated. Cd induced an increase in cellular Cd, Cu, and Zn levels. This results not only in the inhibition of GSH-Px, GRase, CAT, and SOD activities but also in ROS overproduction, oxidative damage, and apoptotic cell death not related to Cu and Zn mechanisms. The thiol groups and GSH levels decreased, whereas the lipid peroxidation and DNA damage increased. The toxicity of Zn results from the imbalance between the inhibition of antioxidant activities and the induction of MT synthesis. The increase in Cu and Zn levels could be explained by the disruption of specific transporter activities, Cd interference with signaling pathways, and metal displacement. Our results suggest that the alteration of Cu and Zn homeostasis is involved in the oxidative stress induced by Cd.     

Influence of ultraviolet radiation, copper, and zinc on microcystin content in Microcystis aeruginosa (Cyanobacteria)

Cyanobacterial toxin production is allied to some unknown trigger resulting in the production of toxins such as microcystin. We hypothesize that microcystins serve as metal ligands to control bioavailability and toxicity of ambient metals. Since ultraviolet radiation (UVR) promotes photo-oxidation of organic metal ligands and influences trace metal bioavailability, the present study aimed to investigate the influence of UVR, Cu, and Zn on specific growth rates, biomass, photosynthetic capacity, and microcystin content in Microcystis aeruginosa. Two toxigenic strains of Microcystis were cultivated using either Lake Erie filtered water or a chemically defined medium, with realistic concentrations of Cu and Zn combined with natural or artificial UVR exposure. Cu was more toxic than Zn on the basis of free ion concentration of trace metals in synthetic medium, although in Lake Erie water total added Zn (10 nM) or Zn plus Cu (10 nM) had a more detrimental effect on biomass and specific growth rate. Natural UVR delivered at 25% ambient levels caused no decrease on the parameters measured (chlorophyll-a, photosynthetic rate), yet artificial levels of UVR (up to 5.9 μmol UVB photons m⁻² s⁻¹) negatively affected biomass and specific growth rate. Cellular levels of microcystin (per unit chlorophyll-a) were concomitant with specific growth rather than being triggered in response either of these stressors (UVR, Zn, and Cu) alone or in combination, in agreement with a purported constitutive production of microcystins.     

The effects of cadmium and the cadmium-zinc interaction on the axenic growth of ectomycorrhizal fungi

Eleven strains of ectomycorrhizal fungi belonging to seven species have been cultured on a cadmium-contaminated growth medium in order to determine their in vitro cadmium tolerance. Four strains were collected from a zinc and cadmium-polluted soil. Radial growth rate was a sensitive parameter to detect Cd toxicity. A wide differential response to Cd was obtained between the individual species. A clear relation between Cd tolerance and site origin of the isolates did not exist, although such a relationship was found when strains are compared within one species. Cd-sensitive and Cd-tolerant strains of Suillus bovinus were studied in more detail. Two isolates were grown on media with combinations of two non-toxic zinc concentrations and three cadmium levels. Adding a higher Zn concentration to the medium resulted in a reduction of the toxic effect of Cd. This antagonistic effect also resulted in a lowered Cd concentration in the mycelium.     

Characterization of the PIB-Type ATPases present in Thermus thermophilus

P(IB)-type ATPases transport heavy metals (Cu(2+), Cu(+), Ag(+), Zn(2+), Cd(2+), Co(2+)) across biomembranes, playing a key role in homeostasis and in the mechanisms of biotolerance of these metals. Three genes coding for putative P(IB)-type ATPases are present in the genome of Thermus thermophilus (HB8 and HB27): the TTC1358, TTC1371, and TTC0354 genes; these genes are annotated, respectively, as two copper transporter (CopA and CopB) genes and a zinc-cadmium transporter (Zn(2+)/Cd(2+)-ATPase) gene. We cloned and expressed the three proteins with 8His tags using a T. thermophilus expression system. After purification, each of the proteins was shown to have phosphodiesterase activity at 65°C with ATP and p-nitrophenyl phosphate (pNPP) as substrates. CopA was found to have greater activity in the presence of Cu(+), while CopB was found to have greater activity in the presence of Cu(2+). The putative Zn(2+)/Cd(2+)-ATPase was truncated at the N terminus and was, surprisingly, activated in vitro by copper but not by zinc or cadmium. When expressed in Escherichia coli, however, the putative Zn(2+)/Cd(2+)-ATPase could be isolated as a full-length protein and the ATPase activity was increased by the addition of Zn(2+) and Cd(2+) as well as by Cu(+). Mutant strains in which each of the three P-type ATPases was deleted singly were constructed. In each case, the deletion increased the sensitivity of the strain to growth in the presence of copper in the medium, indicating that each of the three can pump copper out of the cells and play a role in copper detoxification.     

Maternal and neonatal scalp hair concentrations of zinc, copper, cadmium, and lead: relationship to some lifestyle factors

Postpartum scalp hair samples from 82 term-pregnancy mother/ neonate pairs were analyzed for their concentrations of zinc (Zn), copper (Cu), cadmium (Cd), and lead (Pb), using inductively coupled plasma-mass spectrometry. Maternal and neonatal Zn concentrations had geometric means (and 99% confidence intervals) of 122.5 microg/g (117.9--131.5 microg/g) and 146.9 microg (141.5--156.7 microg/g) respectively. Corresponding Cu values were 18.4 microg/g (17.6--23.8 microg/g) and 6.7 microg/g (6.3--7.6 microg/g). Those of Cd were 0.49 microg/g (0.47--0.69 microg/g) in the mothers and 0.57 microg/g (0.55--0.86 microg/g) in the neonates. For Pb, they were 7.95 microg/g (7.60--9.32 microg/g) and 4.56 microg/g (4.39--5.56 microg/g). Cigarette smoking, despite its relatively low prevalence (19.5%), was associated with lower Zn and higher Cd and Pb concentrations and in lower Zn/Cd and Zn/Pb molar concentration ratios. Smoking also altered interelemental relationships, particularly those of Zn with Cd and Pb and those between Cd and Pb. Smoking frequency appeared to show negative dose-response effects on maternal and neonatal Zn concentrations, Zn/Pb molar concentration ratios, and birth weight. Mothers with a history of oral contraceptive (OC) usage had significantly higher Cu concentrations and lower Zn/Cu molar concentration ratios than non users, with the highest Cu concentrations and lowest Zn/Cu values being associated with third-generation OCs. No similar effects were elicited in the respective neonatal Cu concentrations. Neither alcohol consumption nor prenatal supplementation with iron and/or folic acid had discernible effects on the maternal or neonatal elemental concentrations. The data from this study suggest that in a given population of term-pregnancy mothers and neonates, significant interindividual variations in hair trace element concentrations can occur, irrespective of commonality of general environment, and that lifestyle factors, including cigarette smoking and OC usage history, can be significant contributory factors to such variations. The data are discussed in relation to the effects of smoking-associated exposure to Cd and Pb exposure on Zn availability for placental transfer, as well as on the quantitative maternal Zn supply levels to the fetus resulting from the known tendency of smokers to have lower dietary intakes of Zn.     

重金属诱导拟南芥原生质体DNA 损伤的单细胞凝胶电泳检测

以拟南芥原生质体为实验体系, 研究不同浓度的3种重金属离子对拟南芥原生质体的毒性和DNA损伤的差异。结果表明, 用1-5 mmol.L-1的Zn2+、Cd2+ 和Cu2+分别处理的拟南芥原生质体, 2 小时内活力逐渐下降, 并表现出明显的浓度依赖性;与相同浓度的Cd2+ 和Cu2+ 相比, Zn2+对拟南芥原生质体活力的影响程度较小, 表现出较低的毒性。单细胞凝胶电泳检测发现,用0.1-0.8 mmol .L-1的Zn2+、Cd2+ 和Cu2+ 分别处理拟南芥原生质体30 分钟, 以OTM值表示的原生质体DNA损伤量随重金属离子浓度的增加而递增; 相同浓度(0.5 mmol.L-1)的3种重金属离子相比, Zn2+对原生质体的遗传毒性明显低于Cu2+ 和Cd2+。综合原生质体活力和DNA损伤的单细胞凝胶电泳检测结果, 发现Zn2+对拟南芥原生质体的遗传毒性较低, 而Cd2+ 和Cu2+的遗传毒性较高。本研究建立的拟南芥原生质体实验体系, 结合运用单细胞凝胶电泳技术, 能够快速、灵敏地检测重金属对植物细胞的遗传毒性。     

Induction and degradation of Zn-, Cu- and Cd-thionein in Chang liver cells

Human liver cells (Chang liver) were exposed to 5 micrograms Zn, 2.5 micrograms Cu or 1 microgram Cd/ml in cultured medium. These exogeneous heavy metals were accumulated by the cells and induced de novo synthesis of metallothionein after a 3-h incubation period. The production of Zn-, Cu- or Cd-thionein started in the cells with accumulation of 1 nmol Zn, 0.3 nmol Cu and 0.1 nmol Cd/mg cytosol protein and subsequently the amounts of metal-binding thioneins increased in agreement with the relative amount of metal accumulated in the cytosol over a 24-h period. When cells containing Zn- or Cu-thionein were placed in metal free medium, 70% or 25% of the zinc or copper bound to each original metallothionein was released after 3 h; bound metals decreased to 85% and 65% respectively after 24 h. The disappearance of metal from metallothionein correlated with increases of metal in the medium. On the other hand, 35S-counts incorporated into Zn- and Cu-thionein decreased only to 40% and 15% of the levels in the original metallothionein after 3 h; 35S-counts decreased to 65% and 45%, respectively, after 24 h, indicating that metals bound to metallothionein decreased more quickly than 35S-counts. These results suggest that metals were released from metallothionein and were excreted into the medium. However, 35S- and 109Cd-counts in Cd-thionein changed very little, if at all, in the cells even after a 24-h incubation period. Our data strongly suggest that Zn- and Cu-thionein are degraded in the cells, but that Cd-thionein remains longer than either Zn- or Cu-thionein. When cells containing Zn-thionein were incubated in metal-free medium, Zn-thionein was digested in the cells and peptide fragments ranging about 200-400 daltons were excreted from the cells.     

Phytochelatins as biomarkers for heavy metal stress in maize (Zea mays L.) and wheat (Triticum aestivum L.): combined effects of copper and cadmium

Heavy metal contaminated soils often show increased levels of more than one metal, e.g. copper (Cu), cadmium (Cd), zinc (Zn), lead (Pb) or nickel (Ni). In case such soils are used for crop production, prediction of yield reduction or quality decline due to heavy metals in the soil is inadequate when based only on chemical soil analysis. The use of biomarkers such as phytochelatins (PC), non-protein thiols specifically induced in plants upon exposure to heavy metals, may be an additional tool or diagnostic criterion in heavy metal research and in practice. In the present work, Cu and Cd uptake and induction of PC synthesis are studied with hydroponically grown maize and wheat plants exposed to mixtures of the two metals. We observed a close positive relationship between the concentrations of Cd and PC in the plant shoot material. A decreased shoot concentration of Cd after addition of Cu, due to metal competition at common root absorption sites, coincided with lower shoot PC levels. Also differences in metal uptake and xylary metal transport among the two plant species were reflected in corresponding differences in PC concentration. The observed direct relationship between shoot PC concentration and the degree of metal-induced growth inhibition makes the use of PC promising for the purpose tested for.     

Selenium, iron, copper, and zinc levels and copper-to-zinc ratios in serum of patients at different stages of viral hepatic diseases

Viral hepatic diseases, especially those induced by the hepatitis B virus, can progress into more serious pathological outcomes and eventually to hepatocellular carcinoma. A growing body of evidence indicates that many trace elements play important roles in a number of carcinogenic processes that proceed through various mechanisms. To examine the status of trace elements during the development of hepatic carcinoma, we determined the selenium, iron, copper, and zinc levels and copper-to-zinc ratios in the serum of patients at different stages of viral hepatic disease. We observed significant changes in the selenium, iron copper, and zinc levels in the serum of patients having hepatocellular carcinoma, relative to those of healthy controls (p<0.05). The mean serum copper level in patients with hepatocellular carcinoma was significantly higher than that of the control group. In contrast, the mean selenium, iron, and zinc levels in patients having hepatocellular carcinoma were significantly lower than those of the control group. In addition, the mean zinc level in the serum of patients with hepatic cirrhosis was significantly lower than that of the control group (p<0.05). Moreover, we found markedly elevated Cu: Zn ratios (p<0.05) in patients having hepatic cirrhosis or hepatocellular carcinoma. Our findings imply that the levels of some trace elements, such as selenium, iron, copper, and zinc, and Cu:Zn ratios, might serve as biomarkers for the increased severity of viral hepatic damage.     

Effects of dietary intake of trace metals on tissue contents of sodium and calcium in mice (Mus musculus)

Mice were given either cadmium (Cd), copper (Cu) or zinc (Zn) ad lib, and levels of the metals in the heart, kidneys and liver were measured together with organ contents of sodium (Na) and calcium (Ca). The contents of Cd increased more than 100-fold in all organs, whereas Zn increased by a factor of 2-4. Copper accumulated only in the liver. Cadmium exposure caused the Na and Ca contents in the kidneys to increase by a factor of 2-3, but caused a statistically significant reduction in the Na content of the liver. Cadmium also caused a reduction in the Ca content of the heart. Copper caused a statistically significant doubling of the Na content in the heart, but a significant reduction in the Ca content in this organ. Zinc caused a reduction in the Ca content of the heart. However, the mechanisms behind these effects are not clear. The accumulation of Cd in the kidneys and heart was associated with a gradual change in the Na and Ca levels in these organs, but trace metal accumulation was not associated with any conspicuous changes in the Na or Ca contents in any other organ. Copper was not accumulated in heart, but Cu intake still had marked effects on the Na and Ca contents in this organ. Since the tissue contents of Na and Ca are likely to be physiologically important, these ions may have potential as biomarkers for toxic stress. Since the effects of Cd and Cu differed markedly, the tissue contents of Na and Ca may also be used in a trace metal-specific system of fingerprint biomarkers.     

The Effect of Electrolyte Type and Concentration on the Release of Cd,Cu, Ni,and Zn in Some Contaminated Calcareous Soils

This paper reports the results of desorption experiments of cadmium (Cd), copper (Cu), nickel (Ni), and zinc (Zn) from some contaminated calcareous soils under four electrolyte types (CaCl₂, MgCl₂, NaCl and Na₂SO₄) with different electrolyte concentrations (0.5, 4 and 10 mM). Among electrolytes, CaCl₂ significantly released more metals from soils. There was a negative relationship between total Cu and Zn content and percentage of Cu and Zn released (average of electrolyte concentrations) using CaCl₂ solution, indicating a higher Cu and Zn released when their total content was low. Generally, Cd, Cu, and Zn speciation was affected by both type of electrolytes and their concentrations, whereas Ni speciation stayed mostly stable and was almost unaffected by applied solutions. It can be suggested that beside competition with cations, chloro-complexation is important parameter in Cd release, while CuOH⁺, and to some extent ZnOH⁺ are important species affecting release of Cu and Zn. The distribution coefficient (K_d) values for each metals greatly varied with the types of electrolytes and electrolyte concentration. On the basis of average percentage of metal released under different electrolytes and concentrations the following sequences was found: Cd > Cu > Ni > Zn. The results are important in understanding the mobility of metals under different solutions and indicating that, Cd and Zn soils may pose a higher and lower mobility and ecological risk in contaminated calcareous soils, respectively.     

Trace metals and micronutrients in bone tissues of the red fox <Emphasis Type="Italic">Vulpes vulpes</Emphasis> (L., 1758)

In this study we determined the levels of trace elements (zinc, copper, lead, cadmium and mercury) in three layers of bones of the hip joint (cartilage, compact bone and spongy bone) of 30 red foxes (Vulpes vulpes) from north-western Poland. Concentrations of Cu, Zn, Pb and Cd were determined by atomic absorption spectrophotometry (ICP-AES) in inductively coupled argon plasma using a Perkin-Elmer Optima 2000 DV. Determination of Hg concentration was performed by atomic absorption spectroscopy. In cartilage, compact bone and spongy bone samples from the red fox, median concentrations of the metals studied could be arranged in the following descending series: Zn > Cu > Pb > Cd > Hg, the values ranging from 142 to 0.002 mg/kg dw. There was a significant difference in Cu concentrations, among all the materials analyzed, with much more Cu found in spongy bone than in compact bone. Significant differences were also noted in the case of Hg concentrations in cartilage with compact bone and the spongy bone, and between concentrations of this metal in compact bone and spongy bone. In males, the concentration of Hg in spongy bone was greater than in females. Younger foxes had a higher concentration of this metal in cartilage than adults. The strongest synergistic relationships were observed in spongy bone between the Zn and Cu, Zn and Cd, as well as between Cu and Cd. Statistically significant antagonistic relationships were detected between zinc and lead in compact bone. In addition to monitoring studies conducted on the abiotic environment, an urgent need exists for long-term monitoring of concentrations of heavy metals with long-term effects on living organisms. An important addition is provided by biomonitoring studies on domesticated and free-living mammals, including Canidae.     

Cadmium-induced abnormality in strains of Euglena gracilis: morphological alteration and its prevention by zinc and cyanocobalamin

Euglena gracilis is susceptible to cadmium (Cd) at high concentrations. There are no comparative data on cytotoxicity or abnormality of CdCl₂ to E. gracilis Z and its achlorophyllous mutant SMZ. The present study examined the cytotoxicity of CdCl₂ under continual exposure at levels ranging from sub-ppm to ppm, and assessed the effects of zinc (Zn) or cyanocobalamin (VB₁₂) supplementation on the suppression of Cd-induced abnormal cell proliferation and hypertrophy. With Zn levels restricted to 1 ppm [as Zn⁺⁺], cell growth of both E. gracilis strains was reduced in proportion to Cd concentration. More abnormal cells (hypertrophied, V-shape and starfish-shape) were observed in both strains at sub-ppm levels of Cd. ZnSO₄ supplementation from 2 to 63 ppm significantly suppressed the incidence of Cd-induced abnormality. However, a significant increase in abnormal cells was observed following Zn supplementation at levels of 125 and 250 ppm, which produced remarkable differences in cell morphology. The incidence of abnormal cells varied with supplemented VB₁₂ levels ranging from 4 to 250 ppb in both E. gracilis strains.     

Accumulation of heavy metals in the metal-tolerant fern,Athyrium yokoscense,growing on various environments

The accumulation of copper, zinc and cadmium inA. yokoscense collected from Ashio (copper-contaminated area), Bandai (zinc- and cadmium-contaminated area) and Tama (non-contaminated area), has been investigated. Copper and zinc were accumulated most highly in the root, whilst cadmium was accumulated more in the leaf. The root ofA. yokoscense growing in areas contaminated with metals contained maximum amounts of Cu (5, 989 mg. kg⁻¹ dry weight) and Zn (6,384 mg.kg⁻¹ dry weight), while in the leaf from the Bandai area 164.8 mg Cd.kg⁻¹ dry weight was accumulated. These amounts are far greater than those found inA yokoscense growing on the non-metalliferous habitat (Tama). Twenty five times more zinc and three times more cadmium were found in the dead leaf than in the living leaf. InA. yokoscense growing on soils containing more than 1,000 mg Cu or Zn.kg⁻¹ dry weight, the uptake of copper by the root increased considerably with increasing copper content in the soils, while the uptake of zinc increased only slightly compared with the increase of zinc in the soils.     

Cadmium,iron and zinc interaction and hematological parameters in rat dams and their offspring

The effects of cadmium (Cd) were evaluated in offspring exposed from birth until weaning (neonatal day 0–21) and 4 weeks after exposure cessation focusing on iron (Fe) and zinc (Zn) levels in organs and hematological parameters. Wistar female rats were administered 50 mg Cd/L in drinking water (Cd-exposed) for 4 weeks before mating and during 3 weeks of gestation plus 3 weeks of lactation. Controls were supplied drinking water. At birth, part of Cd-exposed dams’ litters was cross-fostered to control dams (CCd group) and their control litters were cross-fostered to Cd-exposed dams (CdC group). This procedure enabled to discern the effects of gestational, lactational and gestational plus lactational Cd exposure until weaning in F₁ offspring. Elements were analyzed by atomic absorption spectrometry; hematological parameters manually; and histopathological changes by light microscopy. Gestational plus lactational exposure in Cd-exposed dams and their offspring increased Cd and decreased Fe levels, increased Zn in dams and decreased Zn and body weights in 11- and 21-day pups. In 21-day weanling pups, decreased red blood cell (RBC) count, hemoglobin and hematocrit values and increased reticulocytes in peripheral blood were also found with concomitant histopathological finding of extramedullary hematopoiesis in the liver. In cross-fostered pups with gestational exposure (CCd pups), Fe in the liver decreased on day 11 and Zn increased in the kidney on day 21 whereas in pups with lactational exposure (CdC pups) Zn in the brain decreased on day 11 and Fe decreased in the liver and brain on day 21. Regardless of exposure cessation at weaning, in offspring with gestational plus lactational exposure (Cd-exposed) body weights, kidney and brain Fe levels and RBC and hemoglobin remained decreased in blood until puberty. Furthermore Zn levels increased in the liver, kidney and brain. It was concluded that gestational plus lactational Cd exposure caused decreases in Fe and Zn levels and hematotoxic effects in F₁ offspring more pronouncedly than exposure during either gestational or lactational period alone and the adverse effects of maternally mediated Cd exposure continued after exposure cessation into adulthood.