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Effect of pH on metal speciation and resulting metal uptake and toxicity for earthworms.

In the present study, relationships between changes in the solubility and speciation of metals in contaminated soils under different pH regimes and their toxicity to earthworms were investigated. Earthworms (Lumbricus rubellus) were exposed in a laboratory bioassay to metalliferous soils under three pH regimes: Unamended pH, pH lowered by one unit (pH -1), and pH increased by one unit (pH +1). In each soil, total (hot nitric acid-extractable) and 0.01 M CaCl2-extractable metal concentrations were measured and soil pore-water chemistry analyzed to allow metal speciation to be modeled using the Windermere Humic Aqueous Model. Earthworm metal accumulation was determined and toxicity assessed by measuring survival and reproduction and at the molecular level by recording expression of the gene encoding metallothionein-2 (MT-2) using quantitative reverse transcriptase-polymerase chain reaction. Both metal solubility and speciation were found to be highly pH dependent. Metal accumulation in earthworms was influenced by soil concentration and, in some cases (e.g., Cd), by pH. Reproduction was affected (reduced up to 90%) by soil metal level, pH, and their interaction. Relationships between analyzed and calculated Zn concentrations and toxicity and between analyzed and calculated Cd concentrations and tissue accumulation and MT-2 expression were compared by fitting dose-response models and assessing the fit of the data. This analysis indicated that values based on a pH-adjusted free ion concentration best explained toxicity (r2 = 0.82) and accumulation (r2 = 0.54). Expression of MT-2 was, however, poorly correlated (p > 0.05) with all analyzed and modeled soil metal concentrations.

Animals↗

T cell cross-reactivity to heavy metals: identical cryptic peptides may be presented from protein exposed to different metals.

The mechanisms by which metals induce activation of T cells and thus produce allergic and/ or autoimmune reactions are still obscure, and the same is true for the mechanisms that underly T cell cross-reactivity to different heavy metal ions. In the present study, we investigated induction by metals of T cell reactions to cryptic peptides of bovine RNase A. Murine CD4+ T cell hybridomas specific for cryptic RNase peptides presented from Au(III)-treated RNase were used as detection probes. We showed that in vitro treatment of RNase with Pd(II), Pd(IV), Ni(IV), and partially Pt(IV), but not Au(I), Ni(II), or Pt(II), induced presentation of the same cryptic peptides as those presented from Au(III)-treated RNase. That the former heavy metal ions, but not the latter, were able to alter the antigenicity of RNase was reflected by their ability to induce conformational changes of RNase, as detected by circular dichroism spectroscopy. Furthermore, upon immunization against RNase pretreated with these metals, CD4+ T cell hybridomas specific for unidentified cryptic peptides were obtained. In conclusion, "metal-specific" T cell reactions may be directed against cryptic peptides, and metal cross-reactivity in allergic individuals might be due to metal-induced presentation of overlapping, but not identical, panels of cryptic peptides.

Animals↗

Urinary metal and polycyclic aromatic hydrocarbon biomarkers in boilermakers exposed to metal fume and residual oil fly ash.

BACKGROUND: Boilermakers are occupationally exposed to known carcinogens. METHODS: The association of urinary 1-hydroxy-pyrene (1-OHP), a biomarker of polycyclic aromatic hydrocarbon (PAH) exposure, with biomarkers of metal exposure (vanadium, chromium, manganese, nickel, copper, and lead) in boilermakers exposed to metal fume from welding and dust particulates from residual oil fly ash (ROFA) was examined. A repeated measures cohort study was conducted during the overhaul of an oil-fired boiler. Twice-daily urine samples were obtained for 5 days and analyzed for cotinine, 1-OHP, and metals. Generalized estimating equations (GEE) were used to model the multivariate relationship of 1-OHP to the explanatory variables. RESULTS: Metal and 1-OHP levels were determined for 165 urine samples from 20 boilermakers and these levels increased during the workweek. However, the 1-OHP level was not significantly associated with any individual metal level at any time point. CONCLUSION: This suggests that boilermakers were occupationally exposed to PAH and metals, but 1-OHP as a PAH biomarker was unable to serve as a surrogate marker of metal exposure for the metals measured in this study.

Adolescent↗

Metal binding to Bacillus subtilis ferrochelatase and interaction between metal sites.

Ferrochelatase, the terminal enzyme in heme biosynthesis, catalyses metal insertion into protoporphyrin IX. The location of the metal binding site with respect to the bound porphyrin substrate and the mode of metal binding are of central importance for understanding the mechanism of porphyrin metallation. In this work we demonstrate that Zn(2+), which is commonly used as substrate in assays of the ferrochelatase reaction, and Cd(2+), an inhibitor of the enzyme, bind to the invariant amino acids His183 and Glu264 and water molecules, all located within the porphyrin binding cleft. On the other hand, Mg(2+), which has been shown to bind close to the surface at 7 A from His183, was largely absent from its site. Activity measurements demonstrate that Mg(2+) has a stimulatory effect on the enzyme, lowering K(M) for Zn(2+) from 55 to 24 micro M. Changing one of the Mg(2+) binding residues, Glu272, to serine abolishes the effect of Mg(2+). It is proposed that prior to metal insertion the metal may form a sitting-atop (SAT) complex with the invariant His-Glu couple and the porphyrin. Metal binding to the Mg(2+) site may stimulate metal release from the protein ligands and its insertion into the porphyrin.

Bacillus subtilis↗

Heavy metal uptake by Scirpus Littoralis Schrad. from fly ash dosed and metal spiked soils.

Scirpus littoralis is a wetland plant commonly found in Yamuna flood plains of Delhi, India. The ability of Scirpus littoralis to take up and translocate five metals- Mn, Ni, Cu, Zn and Pb from fly ash dosed and metal spiked soils were studied under waterlogged and field conditions for 90 days. Scirpus littoralis accumulated Mn, Ni, Cu, Zn and Pb upto a maximum of 494.92, 56.37, 144.98, 207.95 and 93.08 ppm dry wt., respectively in below ground organs (BO) in 90 days time. The metal content ratios BO/soil (B/S) were higher than shoot/soil ratios (T/S) for all the metals, the highest being for Ni. Metal ratios BO/water (B/W) were also higher than shoot/water (T/W) ratios but the B/W ratio was maximum for Zn. The changes in nutrient status (N, P) in soil water and plants were also studied at interval of 30 days. The Pearson's correlation between metal uptake and N, P uptake were calculated. All the metals except Ni showed negative correlation with nitrogen but they were all non-significant. However, P uptake showed positive correlations with all the metals and all were significant at 1% confidence limit.

Air Pollutants↗

In the absence of catalytic metals ascorbate does not autoxidize at pH 7: ascorbate as a test for catalytic metals.

Trace amounts of adventitious transition metals in buffer solutions can serve as catalysts for many oxidative processes. To fully understand what role these metals may play it is necessary that buffer solutions be 'catalytic metal free'. We demonstrate here that ascorbate can be used in a quick and easy test to determine if near-neutral buffer solutions are indeed 'catalytic metal free'. In buffers which have been rendered free of catalytic metals we have found that ascorbate is quite stable, even at pH 7. The first-order rate constant for the loss of ascorbate in an air-saturated catalytic metal free solution is less than 6 X 10(-7) s-1 at pH 7.0. This upper limit appears to be set by the inability to completely eliminate catalytic metal contamination of solutions and glassware. We conclude that in the absence of catalytic metals, ascorbate is stable at pH 7.

Ascorbic Acid↗

Metal tolerance and biosorption potential of filamentous fungi isolated from metal contaminated agricultural soil.

Heavy metal analysis of agricultural field soil receiving long-term (>20 years) application of municipal and industrial wastewater showed two- to five-fold accumulation of certain heavy metals as compared to untreated soil. Metal-resistant fungi isolated from wastewater-treated soil belonged to genera Aspergillus, Penicillium, Alternaria, Geotrichum, Fusarium, Rhizopus, Monilia and Trichoderma. Minimum inhibitory concentrations (MIC) for Cd, Ni, Cr, Cu, and Co were determined. The MIC ranged from 0.2 to 5 mg ml(-1) for Cd, followed by Ni (0.1-4 mg ml(-1)), Cr (0.3-7 mg ml(-1)), Cu (0.6-9 mg ml(-1)) and for Co (0.1-5 mg ml(-1)) depending on the isolate. Aspergillus and Rhizopus isolates were tested for their metal biosorption potential for Cr and Cd in vitro. Biosorption experiments were conducted with initial metal concentrations of 2, 4, 6 and 8 mM with a contact time of 4 h and wet fungal biomass (1-5 g) at 25 degrees C. Maximum biosorption of Cr and Cd ions was found at 6 mM initial metal concentration. Aspergillus sp.1 accumulated 1.20 mg of Cr and 2.72 mg of Cd per gram of biomass. Accumulation of these two metals by very tolerant Aspergillus sp.2 isolate was at par with relatively less tolerant Aspergillus sp.1 isolate. Rhizopus sp. accumulated 4.33 mg of Cr and 2.72 mg of Cd per g of biomass. The findings indicated promising biosorption of cadmium and chromium by the Rhizopus and Aspergillus spp. from aqueous solution. There is little, if any, correlation between metal tolerance and biosorption properties of the test fungi.

Adaptation, Physiological↗

Heavy metal contamination from mining sites in South Morocco: 2. Assessment of metal accumulation and toxicity in plants.

Metalliferous soils cover a relatively large surface area in Morocco, and up to now no hyperaccumulating plants have been identified on these mining or these industrial sites. The aim of this work was to assess the extent of metal accumulation by plants found in three mining areas in southern Morocco with the ultimate goal of finding metal hyperaccumulating species by using the MetPAD biotest. The biotest helps to obtain information on the selective metal toxicity of aqueous extracts from the plants. A strong metal toxicity, as revealed by the biotest is an indication of a hyperaccumulating plant. Toxicity tests were run concurrently with chemicals analyses of metals in plants and their water extracts. The chemical analyses allow the determination of the hyperaccumulated metal(s). Specimens of the plant species mainly growing on and in the vicinity of the three mines were sampled with their corresponding soils. The results show that all plants analyzed had lower heavy metal content and toxicity despite the relatively very high soil concentrations. A comparison of our results with the criterion used to classify the hyperaccumulator plants indicates that plants we collected from mining sites were hypertolerant but not hyperaccumulators. This was confirmed by transfer factors generally lower than 1. Nevertheless, these tolerant plants species can be used as tools for revegetation for erosion control in metals-contaminated sites (phytostabilization).

Metals, Heavy↗

Heavy metal contamination from mining sites in South Morocco: 1. Use of a biotest to assess metal toxicity of tailings and soils.

Our work was conducted to investigate the heavy metal toxicity of tailings and soils collected from five metal mines located in the south of Morocco. We used the MetPAD biotest Kit which detects the toxicity specifically due to the heavy metals in environmental samples. This biotest initially developed to assess the toxicity of aquatic samples was adapted to the heterogeneous physico-chemical conditions of anthropogenic soils. Contrasted industrial soils were collected from four abandoned mines (A, B, C and E) and one mine (D) still active. The toxicity test was run concurrently with chemical analyses on the aqueous extracts of tailings materials and soils in order to assess the potential availability of heavy metals. Soil pH was variable, ranging from very acidic (pH 2.6) to alkaline values (pH 8.0-8.8). The tailings from polymetallic mines (B and D) contained very high concentrations of Zn (38,000-108,000 mg kg(-1)), Pb (20,412-30,100 mg kg(-1)), Cu (2,019-8,635 mg kg(-1)) and Cd (148-228 mg kg(-1)). Water-extractable metal concentrations (i.e., soil extracts) were much lower but were highly toxic as shown by the MetPAD test, except for soils from mines A, E and site C3 from mine C. The soil extracts from mine D were the most toxic amongst all the soils tested. On this site, the toxicity of soil water extracts was mainly due to high concentrations of Zn (785-1,753 mg l(-1)), Cu (1.8-82 mg l(-1)) and Cd (2.0-2.7 mg l(-1)). The general trend observed was an increase in metal toxicity measured by the biotest with increasing available metal contents in tailings materials and soils. Therefore, the MetPAD test can be used as a rapid and sensitive predictive tool to assess the heavy metal availability in soils highly contaminated by mining activities.

Hydrogen-Ion Concentration↗

Metal availability and soil toxicity after repeated croppings of Thlaspi caerulescens in metal contaminated soils.

Metal phytoextraction with hyperaccumulating plants could be a useful method to decontaminate soils, but it is not fully validated yet. In order to quantify the efficiency of Cd and Zn extraction from a calcareous soil with and without Fe amendment and an acidic soil, we performed a pot experiment with three successive croppings of Thlaspi caerulescens followed by 3 months without plant and 7 weeks with lettuce. We used a combined approach to assess total extraction efficiency (2 M HNO3-extractable metals), changes in metal bio/availability (0.1 M NaNO3-extractable metals and lettuce uptake) and toxicity (lettuce biomass and the BIOMET biosensor). The soil solution was monitored over the whole experiment. In the calcareous soil large Cu concentrations were probably responsible for chlorosis symptoms observed on T. caerulescens. When this soil was treated with Fe, the amount of extracted metal by T. caerulescens increased and metal availability and soil toxicity decreased when compared to the untreated soil. In the acidic soil, T. caerulescens was most efficient: Cd and Zn concentrations in plants were in the range of hyperaccumulation and HNO3-extractable Cd and Zn, metal bio/availability, soil toxicity, and Cd and Zn concentrations in the soil solution decreased significantly. However, a reduced Cd concentration measured in the third T. caerulescens cropping indicated a decrease in metal availability below a critical threshold, whereas the increase of dissolved Cd and Zn concentrations after the third cropping may be the early sign of soil re-equilibration. This indicates that phytoextraction efficiency must be assessed by different approaches in order not to overlook any potential hazard and that an efficient phytoextraction scheme will have to take into account the different dynamics of the soil-plant system.

Biodegradation, Environmental↗

Behavior of Trifolium repens and Lolium perenne growing in a heavy metal contaminated field: Plant metal concentration and phytotoxicity.

The use of a vegetation cover for the management of heavy metal contaminated soils needs prior investigations on the plant species the best sustainable. In this work, behaviors of Trifolium repens and Lolium perenne, growing in a metal-polluted field located near a closed lead smelter, were investigated through Cd, Pb and Zn-plant metal concentrations and their phytotoxicity. In these plant species, metals were preferentially accumulated in roots than in shoots, as follow: Cd>Zn>Pb. Plant exposure to such metals induced oxidative stress in the considered organs as revealed by the variations in malondialdehyde levels and superoxide dismutase activities. These oxidative changes were closely related to metal levels, plant species and organs. Accordingly, L. perenne seemed to be more affected by metal-induced oxidative stress than T. repens. Taken together, these findings allow us to conclude that both the plant species could be suitable for the phytomanagement of metal-polluted soils.

Biodegradation, Environmental↗

Effect of metal spiking on different chemical pools and chemically extractable fractions of heavy metals in sewage sludge.

A laboratory experiment was conducted to study the effect of metal spiking and incubation on some properties and sequentially extractable chemical pools of some heavy metals (F1, two extractions with 0.1 M Sr(NO3)2; F2, one extraction with 1 M NaOAc (pH 5.0); F3, three extractions with 5% NaOCl (pH 8.5) at 90-95 degrees C; F4, three extractions with 0.2 M oxalic acid + 0.2 M ammonium oxalate + 0.1 M ascorbic acid (pH 3.0); and F5, dissolution of sample residue in HF-HClO4 (residual fraction,) and also 1 M CaCl2 and 0.005 M DTPA extractable heavy metals in sewage sludge. Metal spiking and incubation decreased pH and easily oxidizable organic C content of sludge but increased electrical conductivity. Metal spiking and incubation increased F1 fraction of all heavy metals, F2 fraction of Ni, Pb, Cu, and Cd, F3 fraction of Pb, Cu, and Cd, F4 or reducible fraction of Ni, Cu, and Cd and residual fraction of Zn and Pb, but decreased F2 fraction of Zn, F3 of Zn and Ni, F4 fraction of Zn and F5 fraction of Ni, Cu, and Cd. Metal spiking and incubation increased 1 M CaCl2 and 0.005 M DTPA extractable amounts of all heavy metals in sludge except for 0.005 M DTPA extractable Zn, which registered only very marginal decrease.

Biological Availability↗

Estimation of the mass-balance of selected metals in four sanitary landfills in Western Norway, with emphasis on the heavy metal content of the deposited waste and the leachate.

A worst-case simulation of the mass-balance for metals in the waste deposited during 1 year and the levels of cadmium (Cd), lead (Pb), mercury (Hg), chromium (Cr) and iron (Fe) in the leachate was calculated for four sanitary landfills in Western Norway. Estimates of the levels of metal content in mixed municipal solid waste (MSW) were found by using recent literature values calculated in a mass-balance study at a Norwegian waste incinerator plant. Leachate from the landfills were sampled and analyzed monthly during 1 year, and from these measurements the total annual discharge of the selected metals through the leachate was determined. The levels of the measured heavy metals in the leachate were low. For Cd less than 0.06%, for Pb less than 0.01% and for Hg less than 0.02% of the estimated year's deposited mass of metals were leached from the landfills during the year of investigation. The high retention of these metals are most likely due to sulfide precipitation, but also due to the immobile condition of the metals in their original deposited solid state (plastics, ceramics, etc.). The percentage of Cr leached was relatively higher, but less than 1.0% per year. The mass balance of Fe suggests that this element is more mobile under the prevailing conditions. The percentage of Fe leached varied and was estimated to be between 1.9% and 18%. The present study clearly supports the theory that MSW only to a small extent will lead to discharge of metals if deposited at well-constructed sanitary landfills with top layers.

Chromium↗

Changes in mussel biometry on exposure to metals: implications in estimation of metal bioavailability in 'Mussel-Watch' programmes.

The occurrence of changes in flesh and shell weights and in other biometric parameters of mussels, Mytilus galloprovincialis, has been related to different metal levels found in their soft tissues. The effects of clean and Zn-polluted environments and laboratory experiments where Zn-polluted mussels were exposed to sublethal concentrations of Zn, Cu and Cd were investigated. Zinc-polluted mussel shell weights increased significantly after a 51-day depuration period. Exposure of Zn-polluted mussels to Zn or Cd, however, caused a slightly reduced shell growth in comparison with depurating mussel Cu-exposures not causing any reduction in growth. Apart from metal concentrations, metal/shell weight indices have been used to assess metal bioavailability. Metal concentrations recorded in the soft tissues of depurating mussels increased without a source of 'extra' metals, while the Zn/shell-weight index was reduced, as expected from a depuration process, Cu and Cd/shell-weight indices remaining constant. Experimental exposure to Zn, Cu and Cd caused augmented values of Zn, Cu and Cd/shell-weight indices, respectively. These different findings were attributed to changes in flesh weight (related with gamete spawning) that would produce inconsistent estimates of whole metal concentration in soft tissues. Since changes in the tissue composition and in growth rates do not affect Zn/Cu ratios and metal/shell-weight indices, these parameters are proposed as reliable indices of metal bioavailability for 'Mussel-Watch' monitoring programmes. The most sensitive parameter is the metal/shell-weight index, which is, therefore, highly recommended to be used in 'Mussel-Watch' monitoring programmes in order to determine metal bioavailability in seawaters.

Animals↗

Bacterial metal-resistance proteins and their use in biosensors for the detection of bioavailable heavy metals.

We have expressed and purified metal-resistance and metal regulatory proteins from the bacterial determinants of resistance to heavy metals and utilised these in the development of biosensors for heavy metals. Both the metallothionein from the cyanobacterium Synechococcus PCC 7942 and the MerR regulatory protein from transposon Tn501 allow the detection of non-specific metal binding down to 10(-15) M concentrations of Hg(II), Cu(II), Zn(II) and Cd(II) in pure solution. Differential effects of the metals can be detected at both low and high concentrations, and the shape of the capacitance curves may reflect biologically relevant responses of the proteins to metals. Further work is required to establish the relationship between the detected binding of metal and the biological response of the protein, or to provide biosensors of use in the natural environment.

Bacterial Proteins↗

Metal speciation in natural waters with emphasis on reduced sulfur groups as strong metal binding sites.

The proper application of biotic ligand models to predict metal toxicity depends on accurate prediction of metal binding to sites on natural organic matter (NOM) which compete with the 'biotic' ligand for available metal. A hard and soft metal classification along with associated ligand groups (carboxyl, phenolic, amino, sulfidic) are used as a basis to predict metal speciation in the presence of aqueous organic matter. Compilation of conditional metal formation constants (log K') are made for each ligand type using model ligands. Model ligands were chosen to reflect those found in NOM and bio-organic media. Total ligand concentration (L(T)) estimates for different natural settings and log K' values are then used to generate a L(T)-log K' distributions for a specific metal. A plot for Cu(II) gives a similar trend as a compilation of measured data for natural environments. A log K'-L(T) plot for Ag(I) shows a much more discrete binding pattern than for Cu(II). Estimation of speciation of a specific metal in a specific environmental setting and to design speciation and toxicological experiments requires accurate knowledge of the functional groups in NOM.

Animals↗

Advances in biotreatment of acid mine drainage and biorecovery of metals: 1. Metal precipitation for recovery and recycle.

Acid mine drainage (AMD), an acidic metal-bearing wastewater, poses a severe pollution problem attributed to post mining activities. The metals usually encountered in AMD and considered of concern for risk assessment are arsenic, cadmium, iron, lead, manganese, zinc, copper and sulfate. The pollution generated by abandoned mining activities in the area of Butte, Montana has resulted in the designation of the Silver Bow Creek-Butte Area as the largest Superfund (National Priorities List) site in the U.S. This paper reports the results of bench-scale studies conducted to develop a resource recovery based remediation process for the clean up of the Berkeley Pit. The process utilizes selective, sequential precipitation (SSP) of metals as hydroxides and sulfides, such as copper, zinc, aluminum, iron and manganese, from the Berkeley Pit AMD for their removal from the water in a form suitable for additional processing into marketable precipitates and pigments. The metal biorecovery and recycle process is based on complete separation of the biological sulfate reduction step and the metal precipitation step. Hydrogen sulfide produced in the SRB bioreactor systems is used in the precipitation step to form insoluble metal sulfides. The average metal recoveries using the SSP process were as follows: aluminum (as hydroxide) 99.8%, cadmium (as sulfide) 99.7%, cobalt (as sulfide) 99.1% copper (as sulfide) 99.8%, ferrous iron (sulfide) 97.1%, manganese (as sulfide) 87.4%, nickel (as sulfide) 47.8%, and zinc (as sulfide) 100%. The average precipitate purity for metals, copper sulfide, ferric hydroxide, zinc sulfide, aluminum hydroxide and manganese sulfide were: 92.4, 81.5, 97.8, 95.6, 92.1 and 75.0%, respectively. The final produced water contained only calcium and magnesium and both sulfate and sulfide concentrations were below usable water limits. Water quality of this agriculturally usable water met the EPA's gold standard criterion.

Acids↗

Treatment of metal-contaminated water and vertical distribution of metal precipitates in an upflow anaerobic bioreactor.

A lab-scale upflow anaerobic bioreactor filled with granular sludge and cow manure was operated for 140 days to determine the mechanism of metal removal and the vertical distribution of metal precipitates. Heavy metal ions were removed in the order of Cu2+, Cd2+, Zn2+, Fe2+ and Mn2+ with respect to the height in the reactor. The solid phase analysis showed that the heavy metals were mostly precipitated in the form of metal sulfides by sulfate reduction The contents of metal precipitates in the reactor were as follows: (i) Cd and Zn were highest in the bottom, (ii) Fe was highest at the low-middle layer, and (iii) Mn was increased with the height in the reactor. The vertical distribution of metal sulfides in the reactor was directly related to the solubility product (Ksp). Results obtained in this study suggest a feasibility of the application to separate precipitation metal-containing wastewater.

Bacteria, Anaerobic↗