The effects of heavy metal pollution on genetic diversity in zinc/cadmium hyperaccumulator Sedum alfredii populations

The genetic diversity and population structure of seven populations of Sedum alfredii growing in lead/zinc (Pb/Zn) mine spoils or in uncontaminated soils from eastern and southern China were investigated using random amplified polymorphic DNA (RAPD) technology. Four of the sampled sites were heavily contaminated with heavy metals (Zn, Cd, Pb), and extremely high concentrations of Zn, Cd, and Pb were found among these corresponding populations. A significant reduction of genetic diversity was detected in the mining populations. The reduction of genetic diversity could be derived from a bottleneck effect and might also be attributed to the prevalence of vegetative reproduction of the mining populations. Analysis of molecular variance (AMOVA) and the unweighted pair group method with arithmetic mean (UPGMA) tree derived from genetic distances further corroborated that the genetic differentiation between mine populations and uncontaminated populations was significant. Polymorphism with the heavy metal accumulation capability of S. alfredii probably due to the genetic variation among populations and heavy metal contamination could have more impact on the genetic diversity and population structure of S. alfredii populations than geographic distance.     

Effects of inoculation of a plant growth promoting rhizobacterium <Emphasis Type="Italic">Burkholderia</Emphasis> sp. D54 on plant growth and metal uptake by a hyperaccumulator <Emphasis Type="Italic">Sedum alfredii</Emphasis> Hance grown on multiple metal contaminated soil

Batch experiments were designed to characterize a multiple metal resistant bacterium Burkholderia sp. D54 isolated from metal contaminated soils in the Dabaoshan Mine in South China, and a follow-up experiment was conducted to investigate the effects of inoculating the isolate on plant growth and metal uptake by Sedum alfredii Hance grown on soils collected from a heavily contaminated paddy field in Daxing County, Guangxi Zhuang Automounous Region, Southwest China. Our experiments showed that strain D54 produced indole acetic acid (IAA), siderophores, 1-aminocyclopropane-1-carboxylate (ACC) deaminase, and solubilizing inorganic phosphate and solubilized insoluble metal bearing minerals. Bacterial inoculation significantly enhanced S. alfredii biomass production, and increased both shoot and root Cd concentration, but induced little variation in root/shoot Pb concentration and shoot Zn concentration. Despite this, the total shoot and root uptake of Cd, Pb and Zn in S. alfredii inoculated with D54 increased greatly compared to the non-inoculated controls. It was concluded that inoculation with strain D54 could help S. alfredii grow better on metal contaminated soils, produce more biomass, and remove more metals from soil, which implies improved efficiency of phytoextraction from metal contaminated soil. The knowledge gained from the present experiments constitutes an important advancement in understanding of the interaction between plant growth-promoting bacteria and hyperaccumulators with regard to plant ability to grow and remove the multiple heavy metals from soils.     

南京市14种绿化树种对空气中重金属的累积能力

分析了南京化工厂（污染区）和江苏省林科院（对照区）不同绿化树种叶片重金属元素（Pb、Cd和Cu）的含量,揭示了14种绿化树种对3种重金属污染物的累积能力。结果表明,城市绿化树种对大气重金属污染物具有一定的吸收净化能力,并依重金属和树种的不同具有明显差异;同种植物对不同重金属的累积量也有很大差别,其中对Cu的累积量最高;采样区植物叶片重金属含量明显高于对照区;对Pb累积量高的树种有：杨树、广玉兰、女贞和紫叶李; Cd累积量高的树种为杨树;Cu累积量高的树种为构树;累积3种重金属综合能力最强的树种有杨树、构树、雪松、广玉兰、悬铃木、栾树。     

Root foraging for zinc and cadmium requirement in the Zn/Cd hyperaccumulator plant Sedum alfredii

Positive root response to metals may enhance metal accumulation for greater requirement in hyperaccumulators. The effects of spatially heterogeneous Zn/Cd addition on root allocation, metal accumulation, and growth of the Zn/Cd hyperaccumulator Sedum alfredii were assessed in a pot experiment. Young shoots of S. alfredii were grown with or without supplied Zn/Cd. Two concentrations were used of each metal, and each metal concentration had one homogeneous and two heterogeneous treatments. Growth increased by 1.6–3.2 times with the increasing overall dose of Zn/Cd addition, and shoot biomass was positively correlated with shoot Zn/Cd concentration (P?<?0.001). In all heterogeneous treatments, the plants consistently allocated approximately 90% of root biomass to the metal-enriched patches, and shoot Zn/Cd contents were greater than or similar to those in the homogeneous treatment at each metal concentration. Plants in the control treatment showed symptoms of Zn deficiency, although their shoots had Zn concentrations 100-fold higher than the critical deficiency value for most plants. We conclude that S. alfredii has evolved root foraging mechanisms associated with its greater requirements for Zn/Cd. These results could have important implications both for phytoremediation and for investigation of positive role of Cd in higher plants.     

黄河口盐地碱蓬湿地土壤-植物系统重金属污染评价

以黄河口盐地碱蓬湿地为例,评价了淹水和非淹水区湿地表层土壤As、Cd、Cu、Cr、Pb和Zn 6种重金属的污染程度及其在土壤-植物系统中的迁移、富集特征,分析了不同积水深度和土壤理化性质对研究区土壤重金属含量的影响.研究结果表明,与土壤或沉积物质量标准相比,黄河口盐地碱蓬湿地土壤受As和Cd污染最严重,而其它重金属污染较轻;非淹水土壤Cd、Cr和Zn含量高于淹水湿地,而As、Cu和Pb则较低;而且淹水土壤As含量随积水深度增加而呈下降趋势,但积水深度对其他重金属含量的影响不明显.相关性分析结果表明,按照受土壤关键影响因子的不同重金属(除As外)可以分为两类:第一类为Cd、Cr和Zn,这些重金属含量受土壤pH值和盐分影响较大,且相互间存在显著正相关关系,表明它们可能有相同的来源;第二类为Pb和Cu,它们受土壤pH值、盐分和有机质的影响,且Pb和Cu之间存在显著正相关关系.除Cr、Cu和Zn外,重金属在盐地碱蓬的根系内一般不发生显著富集,但绝大多数重金属都表现出地上部分的含量比根系更高的现象.     

Efficiency of selected heavy metals accumulation by Salix viminalis roots

The efficiency of the absorption from the medium and accumulation by plant of ions of the heavy metals depends on many factors including plant age and its genotype. The heavy metals accumulation by 1-, 2- and 3-year-old plants was studied in the aspect of reclamation and revitalization effectiveness of demoted areas. Results of this study answer the question concerning the accumulation of seven heavy metals (Cd, Co, Cr, Cu, Ni, Pb and Zn) by willow cuttings (Salix viminalis) in environments with different concentrations of these elements. Cuttings used were originally grown on a substrate not contaminated with the metals and rich in nutrients. In order to eliminate the effect of soil physicochemical factors, the experiment was carried out using a hydroponic system. Results indicated significant effects of the investigated metal concentrations on their accumulation by willow. The extent of metal accumulation as an indicator of the remediation capacity of willow depended on the age of the cuttings used at planting.     

Phytoextraction of Metals and Rhizoremediation of PAHs in Co-Contaminated Soil by Co-Planting of Sedum Alfredii with Ryegrass (Lolium Perenne) or Castor (Ricinus Communis)

A pot experiment was conducted to investigate the potential for phytoextraction of heavy metals and rhizoremediation of polycyclic aromatic hydrocarbons (PAHs) in co-contaminated soil by co-planting a cadmium/zinc (Cd/Zn) hyperaccumulator and lead (Pb) accumulator Sedum alfredii with ryegrass (Lolium perenne) or castor (Ricinus communis). Co-planting with castor decreased the shoot biomass of S. alfredii as compared to that in monoculture. Cadmium concentration in S. alfredii shoot significantly decreased when grown with ryegrass or castor as compared to that in monoculture. However, no reduction of Zn or Pb concentration in S. alfredii shoot was detected in co-planting treatments. Total removal of either Cd, Zn, or Pb by plants was similar across S. alfredii monoculture or co-planting with ryegrass or castor, except enhanced Pb removal in S. alfredii and ryegrass co-planting treatment. Co-planting of S. alfredii with ryegrass or castor significantly enhanced the pyrene and anthracene dissipation as compared to that in the bare soil or S. alfredii monoculture. This appears to be due to the increased soil microbial population and activities in both co-planting treatments. Co-planting of S. alfredii with ryegrass or castor provides a promising strategy to mitigate both metal and PAH contaminants from co-contaminated soils.     

Lead,cadmium and nickel removal efficiency of white-rot fungus Phlebia brevispora

Widespread of heavy metals contamination has led to several environmental problems. Some biological methods to remove heavy metals from contaminated wastewater are being widely explored. In the present study, the efficiency of a white-rot fungus, Phlebia brevispora to remove different metals (Pb, Cd and Ni) has been evaluated. Atomic absorption spectroscopy of treated and untreated metal containing water revealed that all the metals were efficiently removed by the fungus. Among all the used metals, cadmium was the most toxic metal for fungal growth. Phlebia brevispora removed maximum Pb (97·5%) from 100 mmol l⁻¹ Pb solution, which was closely followed by Cd (91·6%) and Ni (72·7%). Scanning electron microscopic images revealed that the presence of metal altered the morphology and fine texture of fungal hyphae. However, the attachment of metal on mycelia surface was not observed during energy-dispersive X-ray analysis, which points towards the intracellular compartmentation of metals in vacuoles. Thus, the study demonstrated an application of P. brevispora for efficient removal of Pb, Cd and Ni from the metal contaminated water, which can further be applied for bioremediation of heavy metals present in the industrial effluent.     

Phytoremediation of soil co-contaminated with heavy metals and deca-BDE by co-planting of Sedum alfredii with tall fescue associated with Bacillus cereus JP12

Aim

The objective of this study was to develop a remediation strategy for soil co-contaminated with decabromodiphenyl ether (BDE-209) and heavy metals (Cd, Pb and Zn) using co-plantation of the hyperaccumulator plant (Sedum alfredii) with tall fescue (Festuca arundinaceae) associated with a BDE degrader (Bacillus cereus strain JP12).

Methods

A 120-day remediation experiment was conducted under greenhouse conditions. S. alfredii and tall fescue were grown in monoculture and intercropped in artificially contaminated soil. Plant biomass, concentration of polybrominated diphenyl ethers, density of soil bacteria, soil enzyme activity, and the physiological profile of the soil microbial community were determined.

Results and discussion

Inoculation with JP12 significantly increased BDE-209 dissipation in soil. Phytoextraction of metals was also enhanced by JP12 inoculation due to the improved plant growth. Planting of tall fescue significantly enhanced BDE-209 dissipation as compared to that in the bare soil because of the increased soil microbial activity. Tall fescue showed higher Pb phytoextraction efficiency than S. alfredii, but Pb was principally retained in the roots of tall fescue. BDE-209 dissipation and metal phytoextraction were highest when co-planting S. alfredii with tall fescue inoculated with strain JP12. Pyrosequencing analysis revealed that the inoculated JP12 could functionally adapt to the introduced soil, against competition with indigenous microorganisms in soil.

Conclusions

Co-planting of S. alfredii with tall fescue combined with BDE-degrading bacterial strain JP12 is promising for remediation of soil co-contaminated with BDE-209 and metals.     

Metal accumulation and arbuscular mycorrhizal status in metallicolous and nonmetallicolous populations of <Emphasis Type="Italic">Pteris vittata</Emphasis> L. and <Emphasis Type="Italic">Sedum alfredii</Emphasis> Hance

Although Pteris vittata L. and Sedum alfredii Hance have been identified as an As hyperaccumulator and a Zn/Cd hyperaccumulator, respectively, for a few years, variations in metal accumulation among populations and their arbuscular mycorrhizal (AM) status have not been fully explored. Six populations of P. vittata and four populations of S. alfredii from southeast China were investigated. Up to 1,373 As, 680 Pb, 376 Zn, 4.8 Cd, 169 Cu mg kg⁻¹ in fronds of P. vittata and 358 As, 2,290 Pb, 23,403 Zn, 708 Cd, 342 Cu mg kg⁻¹ in shoots of S. alfredii were detected. Constitutive properties of As and Zn hyperaccumulation in metallicolous populations of P. vittata and S. alfredii, respectively, were confirmed. However, Cd hyperaccumulation in S. alfredii varied among populations. The two hyperaccumulators varied in efficiency in taking up other heavy metals. Different metal tolerance strategies adopted by the two hyperaccumulators varied among plant species and metal species. Low to moderate levels of AM colonization in P. vittata (4.2–12.8%) and S. alfredii (8.5–45.8%) were observed at uncontaminated and metal-contaminated sites. The relationship between metal concentrations and AM colonization in the two hyperacumulators was also examined. The abundance of AM fungal spores ranged from 16 to 190 spores per 25 g soil. Glomus microaggregatum, Glomus mosseae, Glomus brohultii and Glomus geosporum were the most common species associated with both P. vittata and S. alfredii. To our knowledge, this is the first report of AM fungal status in rhizosphere of P. vittata and S. alfredii.     

Characterization of coal fly ash and use of plants growing in ash pond for phytoremediation of metals from contaminated agricultural land

Mobilization of heavy metals around coal power plants due to improper disposal of fly ash (FA) and wastewater have led to release of pollutants into the environment. For protection of inimitable natural resources, application of economical and effective technologies is needed such as phytoremediation is cost-effective, ecofriendly and a better option for elimination of metal from contaminated sites. Twelve plant species were sampled from ash dyke of Singrauli and screened for accumulation of metals for this study. Mobilization ratio of metals from soil to plant was evaluated to determine translocation factor. CILLAS analyzer, Raman spectroscopy and SEM-EDX were used for characterization of particle size, functional groups and morphology of fly ash. Results showed mean metal concentrations in contaminated soil for Fe, Mn, Cr, Zn, Ni, Cu, Cd and Pb were 909.4, 60.6, 9.5, 134.8, 13.6, 26.7, 2.9 and 25.4 µg g^?1 respectively. Enrichment factors for soil, root and shoot for a contaminated site were 1.9, 3.8 &; 4.3 for Zn and 2.7, 3.5 &; 3.8 for Cd. Six hyper-accumulators with absorption efficiency >1 viz. I. carnea, S. nigrum, S. munja, T. angustifolia, C. dactylon and P. hysterophorus were identified which may be cultivated successively to reclaim and restore damaged agricultural land.     

Heavy metal accumulation in iron plaque and growth of rice plants upon exposure to single and combined contamination by copper, cadmium and lead

A pot experiment was conducted to evaluate the bioaccumulation of heavy metals and growth response of rice plants after exposure to single and combined contamination by Cu, Cd and Pb. The results showed that the biomass production was not significantly affected by either single or combined treatment of Cu, Cd and Pb. Adding Cu (Cd, or Pb) separately all increased concentrations of the respective element in root and shoot (p < 0.001). In the combined contamination, Pb promoted both root and shoot absorption of Cu and Cd (p < 0.001), and Cu affected Cd and Pb absorption in the root, but Pb concentrations in both root and shoot were not affected by Cd application. The formation of iron plaques varied obviously with soil types. Heavy metal accumulation in iron plaques was induced by the three elements (p < 0.001). Furthermore, the three heavy metals exhibited an interactive relationship as measured by the Cu, Cd, Pb and Fe concentrations in root surface iron plaques. The iron plaques partially inhibited transfer of Pb to root and shoot, but no such effect was observed for Cu and Cd. This research indicates that the interaction among different heavy metal elements is very complex. It is very important to have a clear understanding on the associated mechanism and the consequential impact on plant growth.     

Heavy metals contamination and accumulation in submerged macrophytes in an urban river in China

Deteriorating urban water quality has attracted considerable attention in China. We investigated the contamination levels and distribution of heavy metals (As, Cd, Cu, Ni, Pb, and Zn) in Yuxi River water and sediments, and assessed the heavy metal accumulation capability of five species of submerged macrophytes: Vallisneria natans (Lour.) Hara, Potamogeton pectinatus L., Hydrilla verticillata (L. f.) Royle, Myriophyllum spicatum L., and Potamogeton crispus L. Samples were collected from upstream and downstream locations in different season. The results showed that the levels of heavy metals in the downstream areas were higher than in the upstream areas. Heavy metal concentrations in the river water during the dry seasons were higher than those during the rainy seasons, and the opposite results appeared in sediments and submerged macrophytes. In general, the river was slightly contaminated by heavy metals, and the concentrations of Pb and Ni in this river should serve as a warning, while Cd and Zn pollution in the sediments desperately needs to be removed. Furthermore, Potamogeton pectinatus L. showed a higher accumulation capacity for these metals among the five native submerged macrophytes and could be defined as a hyperaccumulator for Cd. Therefore, the potential use of native aquatic plants in contaminated rivers is worth further exploration.     

Soil nitrogen and microbial biomass carbon dynamics in native forests and derived agricultural land uses in a humid tropical climate of India

Sedum alfredii, a cadmium (Cd) and zinc (Zn) hyperaccumulator at a mine located in Qu Zhou City, Zhejiang Province, China, can accumulate Cd and Zn exceeding 1,000 and 10,000 mg kg^?1, respectively in its shoot (dry weight) when growing under metal-contaminated habitats. Several strains of bacteria were isolated from the rhizosphere of S. alfredii thriving in different Pb/Zn mines in Hunan Province and Zhejiang Province, China, which can resist high levels of heavy metals. Among the different strains isolated, Burkholderia cepacia showed the highest ability in mobilizing Cd and Zn as well as resisting high concentrations of soluble Zn (500 mg L^?1). The soluble Zn concentration in the medium increased from 13 to 72 and 99% (p?<?0.001) after bacterial inoculation in the medium supplemented with insoluble zinc oxide and zinc carbonate, respectively, while pH dropped from 7 to 2.93. The soluble Cd concentration was also increased from 8 to 96% (p?<?0.001), and pH decreased from 7 to 2.65. Short-chain organic acids were also analyzed and the results indicated that oxalic acid, tartaric acid, formic acid and acetic acid had a significant correlation (p?<?0.001) with the concentrations of Cd and Zn being mobilized during the assay. The present results implicated that certain bacteria associated with metal hyperaccumulators could contribute significantly in mobilizing heavy metals, which would enhance the phytoextraction process.     

盐角草在Cd、Pb、Li污染盐土修复中的应用潜力

滩涂和内陆盐碱地是重要的后备土地资源。近年来镉(Cd)、铅(Pb)等土壤重金属和锂(Li)污染对盐碱地开发利用和生产安全造成严重威胁。利用盐生植物修复污染盐土是最经济有效的方法。本研究以盐生植物盐角草SalicorniaeuropaeaL.为材料,采用盆栽方式,通过比较分析盐角草在不同浓度Cd(0-50mmol/L)、Pb(0-50 mmol/L)和Li(0-400 mmol/L)处理下的生长和生理生化指标及离子含量的变化,研究盐角草对3种金属污染物胁迫的耐性及积累特性,以期探讨盐角草在Cd、Pb、Li污染盐土修复中的应用潜力。结果显示,随着Cd、Pb处理浓度升高,盐角草的株高、鲜重和干重均显著下降。低浓度Li(≤20 mmol/L)处理促进盐角草的生长,而高浓度Li(≥20mmol/L)处理则抑制植物生长。盐角草对Cd、Pb、Li的耐受性顺序为Li>Pb>Cd。Cd、Pb、Li胁迫可能降低了盐角草对Na和K的吸收与转运而影响植株的生长。另一方面,盐角草抗氧化酶系统对Cd、Pb、Li胁迫表现出不同的响应机理,多种抗氧化物酶协同作用,抵制Cd、Pb、Li胁迫造成的氧化毒害。盐角草根和地上部分Cd、Pb、Li含量随着处理浓度的升高而增加,其中Cd和Pb的分布特征为根>地上部分,Li的分布特征为地上部分>根。研究结果表明盐角草对Cd、Pb、Li的胁迫均具有较强的耐受性与自我调节能力,且具有富锂特性,具备修复Cd、Pb、Li污染盐土的潜力。本研究为深入研究盐角草耐受Cd、Pb、Li胁迫的机制奠定了基础,揭示了利用盐角草修复高盐碱土壤中Cd、Pb、Li污染的应用潜力。     

Phytotreatment of Sewage Sludge Contaminated by Heavy Metals and Pahs by Co-Planting Sedum Alfredii and Alocasia Marorrhiza

High concentrations of heavy metals and organic pollutants in municipal sewage sludge are key factors limiting its use in agriculture. The objectives of this study were to decrease the heavy metal and polycyclic aromatic hydrocarbon concentrations in sewage sludge by phytotreatment and to determine, in a field experiment, whether co-planting is more effective than using a mono-crop of Sedum alfredii. Four treatments were used in the plot experiment: no sludge, no plants, S. alfredii and co-planting S. alfredii and Alocasia marorrhiza. The results showed that co-planting produced tubers and shoots of A. marorrhiza that were suitable as a safe animal feed and good organic K fertilizer, respectively. Co-planting was more effective than mono-planting at reducing concentrations of total Zn and diethylenetriaminepentaacetic acid (DTPA)-extractable Zn, Cd, and Cu in the sludge. Co-planting decreased the concentrations of DTPA-extractable heavy metals and benzo[a]pyrene (B[a]P) in the sludge significantly compared with the unplanted sludge. Decreases of 87, 75, 85, 31, and 64% were obtained for B[a]P and DTPA-extractable Zn, Cd, Cu, and Pb, respectively, compared with the fresh sludge. These results indicate that co-planting can reduce significantly the environmental risks associated with heavy metals and B[a]P in sewage sludge for further disposal.     

Multi-Wall Carbon Nanotubes Effects on Plant Seedlings Growth and Cadmium/Lead Uptake In Vitro

The effects of multi-wall carbon nanotubes (MWCNTs) on plant growth and Cd/Pb accumulation was investigated on seedlings of three plant species including Brassica napus L., Helianthus annus L. and Cannabis sativa L. The experiment consisted of MWCNTs on three concentration levels (0, 10, 50 mg/L) and 200 μM CdCl₂ or 500 μM Pb(NO₃)₂. MWCNTs application effectively improved root and shoot growth inhibited by Cd and Pb salts. In B. napus, total chlorophyll (Chl) content increased by both MWCNTs 10 and 50 mg/L exposure under cadmium or lead stress. MWCNT 10 mg/L mitigated the deleterious effects of Cd ions on total chlorophyll content of H. annus and C. sativa. Wherease higher concentration of MWCNTs decreased Chl content under either Cd or Pb treatments on sunflower seedlings. MWCNT10 effectivly raised cadmium accumulation in seedlings of all three species. MWCNT10 and 50 mg/L also caused higher Pb accumulation in canola and cannabis seedlings, respectively. Based on the results, it seems that the effects of MWCNTs on growth parameters and heavy metal accumulation in plant seedlings is strongly depends on heavy metal type, MWCNTs concentration and plant species.     

Soil Nitrogen Fertilization Effects on Phytoextraction of Cadmium and Lead by Tobacco (Nicotiana tabacum L.)

A greenhouse experiment using 24 plastic pots filled with 6 kg of Pb- and Cd-contaminated soil was carried out. In all 24 pots, soils were heavy metal–contaminated with 10 mg Cd kg^?1 soil and 500 mg of Pb kg^?1 soil by using CdCl and PbNO₃. Two-month-old tobacco (Nicotiana tabacum L.) plants were used to extract these heavy metals. Results showed that tobacco is able to remove Cd and Pb from contaminated soils and concentrate them in its harvestable part, that is, it could be very useful in phytoextraction of these heavy metals. Increasing additions of ammonium nitrate to soil (50, 100, and 150 mg N kg^?1 soil) significantly (p ≤ .05) increased aboveground Cd and Pb accumulation during a 50-day experimental period, whereas increasing additions of urea to soil (50 and 100 mg N kg^?1 soil) did not show these effects at the same significance levels. Increasing additions of ammonium nitrate to soil shows as dry matter increases, both accumulated Cd and accumulated Pb also increase when tobacco plants are growing under Pb- and Cd-contaminated soil conditions. Higher Pb concentrations depress Cd/Pb ratios for concentrations and accumulations, suggesting that Pb negatively affects Cd concentration and/or accumulation.     

Epidermal uptake of Pb,Cd, and Zn in tubificid worms

Summary The epidermal uptake of Pb, Cd, and the essential element Zn by tubificid worms was investigated. The animals were starved and contaminated via the water column at 4°C and 25°C. Atomic absorption spectroscopy (AAS), energy-dispersive X-ray microanalysis (EDX), laser induced mass analysis (LAMMA), electron microscopy, and the sulfide-silver method at the electron microscopical level were used. The sulfidesilver method revealed many reaction products indicating heavy metals in membrane-limited lysosomal structures in epidermal cells of contaminated Limnodrilus udekemianus. In these lysosomes, which were particularly abundant in the epidermis of the hind end, Pb, Cd, and Zn were detected by LAMMA and EDX analysis. Beside this distinct occurrence of the three elements a uniform pattern of reaction products was found in epidermal cells after contamination with Cd and Zn, but not after treatment with Pb. As shown by atomic absorption spectroscopy, Cd was the most enriched element at 25°C, followed by Pb and Zn. Simultaneous application of Zn reduced the uptake of Cd, whereas the uptake of Pb was increased. At 4° C uptake of Pb and Cd was slower than at 25° C, while Zn uptake was blocked completely. This shows that heavy metal uptake can be strongly dependent on environmental conditions, which has to be taken into account if animals are used as monitor organisms for heavy metal pollution in the environment.