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.     

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.     

Performance of bioaugmentation-assisted phytoextraction applied to metal contaminated soils: a review

Bioaugmentation-assisted phytoextraction is a promising method for the cleaning-up of soils contaminated by metals. Bacteria mainly Plant Growth Promoting Rhizobacteria (PGPR) and fungi mainly Arbuscular Mycorrhizal Fungi (AMF) associated with hyperaccumulating or non-hyperaccumulating plants were analyzed on the basis of a bioprocess engineering approach (concentration and amount of metals extracted by plants, translocation and bioconcentration factor, and plant biomass). In average bioaugmentation increased metals accumulated by shoots by a factor of about 2 (metal concentration) and 5 (amount) without any obvious differences between bacteria and fungi. To optimize this process, new relevant microorganism-plant associations and field scale experiments are needed along with a common methodology for the comparison of all experiments on the same basis. Recommendations were suggested concerning both the microbial-plant selection and the implementation of bioaugmentation to enhance the microbial survival. The use of microbial consortia associated with plant was discussed notably for multi-contaminated soils.     

Heavy metal concentrations in soils and plant accumulation in a restored manganese mineland in Guangxi, South China

Heavy metal contamination of metal-mined soils is a widespread problem in China. In the restored (over 20 years) Lipu manganese mineland, 36 plant species from 22 families were found colonizing, some of which were planted agronomic ones. Heavy metal concentrations in tailings were very high. Minesoils were basically unpolluted, but soils in the remaining mining area and in the vicinity of tailings dumps were polluted by Cd and Mn. Measurements of metal contents in dominant plants showed they were close to those of other mineland plants. Plants tended to have a higher Cd accumulation (as reflected by Biological Accumulation Coefficient) from soil, but have a higher Mn translocation (as indicated by Biological Transfer Coefficient) to aboveground parts. The Chinese chestnut and sugarcane cultivated on the reclaimed mineland were not safe for human consumption, and this agricultural restoration pattern should be carefully reconsidered.     

Potential of Hemidesmus indicus for phytoextraction of lead from industrially contaminated soils

Many sites in the industrial region of Kattedan near Hyderabad, Andhra Pradesh (AP), India are contaminated with high concentrations of lead. The use of plants to remove toxic metals from soils (phytoremediation) is emerging as a potential strategy for cost-effective and environmentally sound remediation of contaminated soils. We studied remediation of soils contaminated with lead using a lead hyperaccumulating plant, Hemidesmus indicus. The ability of this plant to accumulate lead in shoots and roots was studied with pot experiments. The results showed that accumulation was maximum in roots for the first 1-3 weeks and later for a contact period of three months, the accumulation rate was maximum in shoots. In addition we used various chelating agents such as EDTA, HEDTA, DTPA and CDTA to determine the best chemical modifier for efficient lead removal from contaminated soils. The effect on lead accumulation of plant in the presence of various metal co-ions was also studied. An attempt was made for the decontamination of lead from five different "Real-life" soils of Kattedan using H. indicus.     

Evaluation of metal mobility, plant availability and immobilization by chemical agents in a limed-silty soil

Metal-contaminated soils in the vicinity of industrial sites become of ever-increasing concern. Diagnostic criteria and ecological technologies for soil remediation should be calibrated for various soil conditions; actually, our knowledge of calcareous soil is poor. Silty soils near smelters at Evin (Pas de Calais, France) have been contaminated by non-ferrous metal fallout and regularly limed using foams. Therefore, the mobility, bioavailability, and potential phytotoxicity of Cd, Pb and Zn, were investigated using single soil extractions (i.e. water, 0.1 n Ca(NO(3))(2), and EDTA pH 7), and vegetation experiments, in parallel with a biological test based on (iso)-enzymes in leaves and roots, before and following soil treatment with chemical agents, i.e. Thomas basic slags (TBS), hydrous manganese oxide (HMO), steel shots (ST) and beringite. No visible toxicity symptoms developed on the above-ground parts of ryegrass, tobacco and bean plants grown in potted soil under controlled environmental conditions. Cd, Zn and Pb uptake resulted in high concentrations in the above-ground plant parts, but the enzyme capacities in leaves and roots, and the peroxidase pattern indicated that these metal concentrations were not phytotoxic for beans as test plants. The addition of chemical agents to the soil did not increase biomass production, but treatment with either HMO, ST or beringite markedly decreased the mobility of Cd, Zn and Pb. These agents were proven to be effective in mitigating the Cd uptake by plants. HMO and ST decreased either Pb or Zn uptake by ryegrass. TBS was effective in lowering Pb uptake by the same species. Beringite decreased Cd uptake by beans. If fallout could be restricted, the metal content of food crops in this area should be lowered by soil treatment. However, the differences in Cd uptake between plant species were not suppressed, regardless of the type of agents applied to the soil.     

Bioaccumulation of heavy metals in the earthworms Lumbricus rubellus and Aporrectodea caliginosa in relation to total and available metal concentrations in field soils

The aim of this study was to determine important metal pools for bioaccumulation by the earthworms Lumbricus rubellus and Aporrectodea caliginosa in soils with high binding capacity. Cd, Cu and Zn concentrations in soil, pore water and CaCl(2) extracts of soil, in leaves of the plant species Urtica dioica and in earthworms were determined at 15 field sites constituting a gradient in metal pollution. Variations in the Cu and Cd concentrations in L. rubellus and Cu concentrations in A. caliginosa were best explained by total soil concentrations, while variation in Cd concentration in A. caliginosa was best explained by pore water concentrations. Zn concentrations in L. rubellus and A. caliginosa were not significantly correlated to any determined variable. It is concluded that despite low availability, earthworms in floodplain soils contain elevated concentrations of Cu and Cd, suggesting that uptake takes place not only from the soluble metal concentrations.     

Heavy metal accumulation in Halimione portulacoides: intra- and extra-cellular metal binding sites

Salt marsh plants can sequestrate and inherently tolerate high metal concentrations found in salt marsh sediments. This work intended to understand the Halimione portulacoides (L.) Aellen strategies to prevent metal toxicity, by investigating the metal location in different plant organs and in the cell. A sequential extraction was performed on leaves, stems and roots of H. portulacoides in order to determine and compare the metal (Zn, Pb, Co, Cd, Ni and Cu) concentration in several fractions of the plant material (ethanolic, aqueous, proteic, pectic, polissacaridic, lenhinic and cellulosic). This study shows that all plant organs of H. portulacoides mostly retain metals in the cell wall (65% is the average for all studied metals stored in the root cell wall, 55% in the stems and 53% in the leaves), and the metal content in the intracellular compartment is much lower (21% in roots, 25% in stems and 32% in leaves). High levels of heavy metal in the sedimentary environment do not cause toxicity to H. portulacoides, because H. portulacoides immobilizes them in different cell compartments (cell wall+proteic fraction+intracellular) outside key metabolic sites.     

Survival strategies of plants associated with arbuscular mycorrhizal fungi on toxic mine tailings

A field survey of metal concentrations and arbuscular mycorrhizal (AM) components of plants growing on five mining sites was conducted in Chenzhou City, Hunan Province, Southern China and a control site in Hong Kong. Significant differences were observed in the average concentrations of total heavy metals (Pb, Zn, Cu, Cd) and one metalloid (As) in contaminated soils compared with the control site. Gramineae and Compositae were the dominant plant families growing on mine tailings, with Chrysanthemum moritolium (common chrysanthemum), Cynodon dactylon (Bermuda grass), Miscanthus florodulus (Sword grass) and Pteris vittata (Ladder brake fern) commonly found at all sites. AM fungal colonization was detected in most of the plants. Comparing the four common plant species, three components of mycorrhizal colonization (arbuscules, vesicles and coiled hyphae) were found in the roots of C. dactylon and P. vittata growing at Do Shun Long (DSL) mine site. Concentrations of As in fronds were 24-fold higher than in roots of P. vittata with the highest mycorrhizal colonization rate (73%) among all sampling sites. Extensive mycorrhizal colonization (85%) was also recorded in the roots of C. dactylon with As accumulation 57 times higher than in shoots. The four common plants found in metal contaminated sites had developed different strategies for survival in the contaminated sites with the aid of indigenous AM fungi.     

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.     

Direct and indirect effects of metal contamination on soil biota in a Zn-Pb post-mining and smelting area (S Poland)

Effects of metal contamination on soil biota activity were investigated at 43 sites in 5 different habitats (defined by substratum and vegetation type) in a post-mining area. Sites were characterised in terms of soil pH and texture, nutrient status, total and exchangeable metal concentrations, as well as plant species richness and cover, abundances of enchytraeids, nematodes and tardigrades, and microbial respiration and biomass. The concentrations of total trace metals were highest in soils developed on mining waste (metal-rich dolomite), but these habitats were more attractive than sandy sites for plants and soil biota because of their higher content of organic matter, clay and nutrients. Soil mesofauna and microbes were strongly dependent on natural habitat properties. Pollution (exchangeable Zn and Cd) negatively affected only enchytraeid density; due to a positive relationship between enchytraeids and microbes it indirectly reduced microbial activity.     

Chelate-induced phytoextraction of metal polluted soils with Brachiaria decumbens

Chelate-induced phytoextraction with high biomass plant species has been proposed for the clean-up of heavy metal polluted soils. In the current work, the effect of the application of two different chelating agents, i.e. EDTA and EDDS, on the metal phytoextraction capacity of Brachiaria decumbens was studied. Although EDTA was, in general, more effective in soil metal solubilization, EDDS, a chelate less harmful to the environment, was more efficient inducing metal accumulation in B. decumbens shoots than EDTA. Indeed, in a moderately heavy metal polluted soil, EDDS caused a 2.54, 2.74 and 4.30-fold increase in Cd, Zn, and Pb shoot metal concentration, respectively, as compared to control plants. In this same soil, EDTA caused a 1.77, 1.11 and 1.87-fold increase in Cd, Zn, and Pb shoot metal concentration, respectively, as compared to control plants. EDDS was also more effective than EDTA in stimulating the translocation of metals from roots to shoots. B. decumbens plants were able to grow in the metal polluted soils showing no visible symptoms of phytotoxicity, which suggests their metal tolerance. Finally, B. decumbens, a fast-growing, high biomass, aluminum tolerant plant species, that has a well-established agronomic system, fulfills most of the requirements for chemically-induced phytoextraction.     

Speciation of Cd and Zn in contaminated soils assessed by DGT-DIFS, and WHAM/Model VI in relation to uptake by spinach and ryegrass

A pot experiment was carried out to investigate the impact of Cd and Zn extractability in soil and speciation in pore water of industrial contaminated soils, on metal concentration in a metal sensitive species like spinach (Spinacia oleracea) and a more metal tolerant species like Italian ryegrass (Lolium multiflorum). For chemical speciation of Cd and Zn in pore water, WHAM/Model VI version 6.0 was used. The DGT technique was used to determine the effective concentration, C(E), of Cd and Zn in soils. The free ion activity in pore water correlated well with the contents in plants, and there was a linear relationship between the C(E) values and the concentration of Cd and Zn in both spinach and ryegrass in the non-toxic range. However, the C(E) values usually overestimated the plant contents when plants, particularly the spinach plants, were subjected to toxic concentration in the pore water. Metal uptake decreased in plants affected by toxicity, whereas metal binding to the Chelex resin did not. Thus, we found no linear relationship between the C(E) and metal contents in spinach, whereas a linear relationship was found between C(E)-Zn and the Zn concentration in ryegrass (r2=0.96, p<0.001). For Cd in ryegrass this relationship was weak (r2=0.53, p=0.18). This study indicates that the transport of metals from labile metal pools to the DGT-resin is linearly related to plant uptake only when plants are growing well, and that the applicability of DGT as an indicator for plant uptake seems species dependent.     

Evaluating three trace metal contaminated sites: a field and laboratory investigation

Selecting guidelines to evaluate elevated metals in urban brownfields is hindered by the lack of information for these sites on ecosystem structure and function. A study was performed to compare three trace metal-contaminated sites in the metropolitan Montreal area. The goal was to obtain an idea of the organisms that may be present on urban brownfields and to measure if elevated metals alter the presence and activity of the indigenous biota. Field and laboratory studies were conducted using simple methodologies to determine the extent to which microbial activity affected by trace metal content, to assess diversity of plant and soil invertebrate communities and to measure phytoaccumulation of trace metals. It was found that microbial activity, as measured by substrate-induced respiration (SIR) and nitrification, was not affected by the levels of soil Cd, Cu, Ni, Pb and Zn recorded on the sites. Seven of the 12 invertebrate groups collected were sampled on soils with similar Cd, Cu, Ni, Pb and Zn concentrations. Diversity of plant species increased as a function of the length of time the sites had been inactive. Levels of metals in plant tissue were influenced by soil characteristics and not by total soil Cd, Cu, Ni, Pb and Zn.     

Heavy metal tolerance of rice plants (Oryza sativa L.) to some metal oxides at the critical levels

The toxicity of metal oxides (CdO, ZnO, PbO) were compared with each other and the critical levels of metal uptake by rice plants were determined. The order of metal toxicity to rice plants is CdO greater than ZnO greater than PbO. The highest concentration obtained for unpolished rice was 4.97 micrograms/g at the Cd 10,000 ppm in soil. This values is higher than every other we have seen in the reports for treatment with CdO. We are able to find out that the concentration of 10,000 ppm Cd in the form of CdO in the critical one towards rice plant. By contrast, such damage was not observed at even higher levels of ZnO and PbO were considered to have low toxicity toward rice plant. Also, a negative correlation between Zn or Cu accumulation in rice plants and Cd concentration in soil was found.     

Effects of mycorrhizae and other soil microbes on revegetation of heavy metal contaminated mine spoil

The effects of mycorrhizal fungi and other soil microorganisms on growth of two grasses, Andropogon gerardii Vitm. and Festuca arundinacea Schreb., in heavy metal-contaminated soil and mine tailings were investigated. A. gerardii is highly dependent on mycorrhizal fungi in native prairie, while F. arundinacea is a facultative mycotroph and relies on mycorrhizal symbiosis only in extremely infertile soils. Regardless of microbial amendments, neither plant species was able to establish and grow in the mine tailings. Both plant species grew in the moderately contaminated or non-contaminated soils, although A. gerardii grew in these soils only when mycorrhizal. Other soil microbes significantly improved growth of A. gerardii only in uncontaminated soil, but to a lesser extent than mycorrhizae. Although F. arundinacea was more highly colonized by mycorrhizal fungi than A. gerardii, neither microbial amendment affected growth of fescue in any soil. In several treatments mycorrhizal fungi adapted to uncontaminated soil stimulated plant growth more than mycorrhizae adapted to the moderately contaminated soil. However, mycorrhizal fungi adapted to contaminated soil did not increase the productivity of plant growth in contaminated soil more than fungi adapted to uncontaminated soil. A. gerardii plants inoculated with mycorrhizal fungi retained more Zn in roots than in shoots, confirming earlier reports that mycorrhizal fungi alter the translocation pattern of heavy metals in host plants. In contrast, mycorrhizae did not affect translocation patterns in F. arundinaceae, suggesting that the mycorrhizal dependence of a plant species is correlated with the retention of metals in roots. The correlation between mycorrhizal dependence of a plant species and mycorrhizal alteration of translocation pattern may also explain the inconsistent reports of mycorrhizal effects on translocation of heavy metals in plants. Plant response to mycorrhizal symbiosis may therefore provide a useful criterion for the selection of the plant species to be used in revegetation of contaminated sites.     

Trace metal speciation and bioavailability in urban soils

Urban soils often contain concentrations of trace metals that exceed regulatory levels. However, the threat posed by trace metals to human health and the environment is thought to be dependent on their speciation in the soil solution rather than the total concentration. Three inactive railway yards in Montréal, Québec, were sampled to investigate the speciation and bioavailability of Cd, Cu, Ni, Pb and Zn. Soil solutions were obtained by centrifuging saturated soil pastes. In the soil solutions, up to 59% of the dissolved Cd was in its free ionic form. For Cu, Pb and Zn, organic complexes were the predominant species. Over 40% of Ni was present as inorganic complexes if the solution pH exceeded 8.1. Multiple regression analyses showed that pH and total metals in soil were significantly correlated with the activities of free metal ions, except for Cd(2+), which only had a weak correlation with soil pH. Free, dissolved and total soil metals were tested for their ability to predict metal uptake by plants in the field. However, none of these metal pools were satisfactory predictors. The results indicated that in these urban soils, trace metals were mainly in stable forms and bioavailability was extremely low.     

Effect of new soil metal immobilizing agents on metal toxicity to terrestrial invertebrates

Application of 5% (w:w) novel metal immobilizing agent reduced the water soluble, the calcium chloride extracted as well as the pore water concentration of zinc in soils from Maatheide, a metal contaminated site in the northeast of Belgium. Addition of the metal immobilizing agents also eliminated acute toxicity to the potworm Enchytraeus albidus and the earthworm Eisenia fetida and chronic toxicity to the springtail Folsomia candida. Cocoon production by E. fetida, however, was still adversely affected. These differences may be explained by the species dependent routes of metal uptake: F. candida is probably mainly exposed via pore water while in E. fetida dietary exposure is probably also important. From these results it is clear that organisms with different exposure routes should be used simultaneously to assess the environmental risk of metal contaminated soils.     

The effects of multiple metal contamination on ectomycorrhizal Scots pine (Pinus sylvestris) seedlings

Experiments were conducted to investigate the effects of single and multiple metal contamination (Cd, Pb, Zn, Sb, Cu) on Scots pine seedlings colonised by ectomycorrhizal (ECM) fungi from natural soil inoculum. Seedlings were grown in either contaminated field soil from the site of a chemical accident, soils amended with five metals contaminating the site, or in soil from an uncontaminated control site. Although contaminated and metal-amended soil significantly inhibited root and shoot growth of the Scots pine seedlings, total root tip density was not affected. Of the five metals tested in amended soils, Cd was the most toxic to ECM Scots pine. Field-contaminated soil had a toxic effect on ECM fungi associated with Scots pine seedlings and caused shifts in ECM species composition on ECM seedlings. When compared to soils amended with only one metal, soils amended with a combination of all five metals tested had lower relative toxicity and less accumulation of Pb, Zn and Sb into seedlings. This would indicate that the toxicity of multiple metal contamination cannot be predicted from the individual toxicity of the metals investigated.