Complementary assessment of As, Cu and Zn environmental availability in a stabilised contaminated soil using large-bore column leaching, automatic microcolumn extraction and DGT analysis.

Soil pollution with trace elements is a concerning issue worldwide. Monitoring of soil pollution and remediation protocols need still from valid complementary analytical approaches able to detect changes in speciation and lability of metals in soils (e.g. stabilization or mobility). In this work, we compare three different analytical approaches to assess potential changes in environmental availability of Cu, As and Zn in a Mediterranean polluted soil that was amended with different combinations of iron sulphate and alkaline paper sludge waste. The studied methods were: (i) a standard large-bore flow-through column system (macrocolumn), (ii) an automatic dynamic flow-through microcolumn extraction system, and (iii) the diffusive gradients in thin gels technique (DGT). The three analytical approaches tested showed immobilization of Zn and Cu in contaminated mine soils after co-application of paper sludge and iron sulphate, but they differ quantitatively in terms of As mobility. Interconversion between oxidation states of inorganic As is observed to occur to a larger extent in macrocolumn. Because this may only occur in very specific Mediterranean scenarios (i.e. organic matter application to intermittently flooded mine soils), macrocolumn extraction procedures might not appropriately mimic the environmental availability of As in soils with organic amendments (e.g., paper sludge waste). Microcolumn leaching is the fastest screening tool to ascertain the efficiency of chemical amendments, but DGT is a good alternative with less technical demands.

Synergic use of chemical and ecotoxicological tools for evaluating multi-contaminated soils amended with iron oxides-rich materials.

Abandoned waste piles from ancient mining activities are potential hot spots for the pollution of the surrounding areas. A pot experiment was carried out to check the potential toxicity of the dumping material present in one of these scenarios, and several amendments were tested to attenuate the spread of the contamination events. The waste material had an acid pH and a large total concentration of As and Cu. A dose-response experiment was performed with this material following OCDE 208 test. A proportion 90:10 uncontaminated soil: dumping material (% w/w) was selected for the following experiment, in order to surpass the amount of dumping material that caused 50% reduction in plant growth. Pots were filled with the 90:10 mixture, planted with seeds of Brassica napus and amended with the following materials: three iron oxides of Bayoxide® E33 series, iron (II) sulphate in combination with de-inking paper sludge (Fe+PS), iron oxide-rich rolling mill scale (ROL) and iron oxide-rich cement waste (CEM). Amendment effectiveness evaluation was based on chemical and biological assays: extractable trace element concentration, soil enzymatic activities, inhibition of light emission of V. fischeri and Anabaena sp., B. napus L. fresh weight and screening test for emergence of B. napus L. seedlings. Amendments E33HCF and Fe+PS were the most effective in reducing extractable As and Zn concentration. B. napus weight and dehydrogenase and ß-glucosidase activities were positively increased with the two above mentioned treatments but they triggered more toxic effects for V. fischeri luminescence. E33P treatment was the only in which the EC50 was higher than in the control. Anabaena sp. was less sensitive than V. fischeri as its luminescence was not hampered by any treatment. Trace element concentration did not significantly affect the failure in seed emergence. E33HCF and Fe+PS could act as proper amendments as they decreased extractable As and Zn. Further, plant fresh weight, enzymatic activities and some of the bioassays identified the latter treatments as the best ones among those tested here to this type of multi-contaminated soil.

Assessing the ecotoxicological effects of long-term contaminated mine soils on plants and earthworms: relevance of soil (total and available) and body concentrations.

The interactions and relevance of the soil (total and available) concentrations, accumulation, and acute toxicity of several essential and non-essential trace elements were investigated to determine their importance in environmental soil assessment. Three plant species (T. aestivum, R. sativum, and V. sativa) and E. fetida were simultaneously exposed for 21 days to long-term contaminated soils collected from the surroundings of an abandoned pyrite mine. The soils presented different levels of As and metals, mainly Zn and Cu, and were tested at different soil concentrations [12.5, 25, 50, and 100% of contaminated soil/soil (w/w)] to increase the range of total and available soil concentrations necessary for the study. The total concentrations in the soils (of both As and metals) were better predictors of earthworm uptake than were the available concentrations. In plants, the accumulation of metals was related to the available concentrations of Zn and Cu, which could indicate that plants and earthworms accumulate elements from different pools of soil contaminants. Moreover, Zn and Cu, which are essential elements, showed controlled uptake at low concentrations. The external metal concentrations predicted earthworm mortality, whereas in plants, the effects on growth were correlated to the As and metal contents in the plants. In general, the bioaccumulation factors were lower at higher exposure levels, which implies the existence of auto-regulation in the uptake of both essential and non-essential elements by plants and earthworms.

Amendment application in a multi-contaminated mine soil: effects on soil enzymatic activities and ecotoxicological characteristics.

Several amendments were tested on soils obtained from an arsenopyrite mine, further planted with Arrhenatherum elatius and Festuca curvifolia, in order to assess their ability to improve soil's ecotoxicological characteristics. The properties used to assess the effects were: soil enzymatic activities (dehydrogenase, ß-glucosidase, acid phosphatase, urease, protease and cellulase), terrestrial bioassays (Eisenia fetida mortality and avoidance behaviour), and aquatic bioassays using a soil leachate (Daphnia magna immobilisation and Vibrio fischeri bioluminescence inhibition). The treatment with FeSO4 1 % w/w was able to reduce extractable As in soil, but increased the extractable Cu, Mn and Zn concentrations, as a consequence of the decrease in soil pH, in relation to the unamended soil, from 5.0 to 3.4, respectively. As a consequence, this treatment had a detrimental effect in some of the soil enzymatic activities (e.g. dehydrogenase, acid phosphatase, urease and cellulase), did not allow plant growth, induced E. fetida mortality in the highest concentration tested (100 % w/w), and its soil leachate was very toxic towards D. magna and V. fischeri. The combined application of FeSO4 1 % w/w with other treatments (e.g. CaCO3 1 % w/w and paper mill 1 % w/w) allowed a decrease in extractable As and metals, and a soil pH value closer to neutrality. As a consequence, dehydrogenase activity, plant growth and some of the bioassays identified those as better soil treatments to this type of multi-contaminated soil.

Risk assessment of an abandoned pyrite mine in Spain based on direct toxicity assays.

This research reports the risk assessment of an abandoned pyrite mine using direct toxicity assays of soil and groundwater samples taken at the site. The toxicity of As and heavy metals from mining soils to soil and aquatic organisms was studied using the Multispecies Soil System (MS-3) in soil columns. Ecotoxicological assessment was performed with soil samples diluted with a control soil at concentrations of 12.5, 25, 50 and 100% test soil/soil (w/w). In this way, changes in the mobility and bioavailability of soil contaminants due to changes in geochemical soil properties via soil dilution were studied. The toxicity of water samples was tested on algae and Daphnia magna. The assessment of the mining area indicated that the current presence of As and heavy metals at the site may cause injuries to soil and aquatic organisms in the entire research area. Moreover, this investigation demonstrated that changes in geochemical conditions can increase the availability of arsenic and, consequently, the environmental risk of these soils. A good correlation was not found between toxicity parameters and the concentrations of soil contaminants based on total and extracted element concentrations. This finding reinforces the usefulness of direct toxicity assays for evaluating environmental risk.

Responses of Noccaea caerulescens and Lupinus albus in trace elements-contaminated soils.

Plants exposed to trace elements can suffer from oxidative stress, which is characterised by the accumulation of reactive oxygen species, alteration in the cellular antioxidant defence system and ultimately lipid peroxidation. We assessed the most-appropriate stress indexes to describe the response of two plant species, with different strategies for coping with trace elements (TEs), to particular contaminants. Noccaea caerulescens, a hyperaccumulator, and Lupinus albus, an excluder, were grown in three soils of differing pH: an acidic soil, a neutral soil (both contaminated mainly by Cu, Zn and As) and a control soil. Then, plant stress indicators were measured. As expected, N. caerulescens accumulated higher levels of Zn and Cd in shoots than L. albus, this effect being stronger in the acid soil, reflecting greater TE solubility in this soil. However, the shoot concentrations of Mn were higher in L. albus than in N. caerulescens, while the As concentration was similar in the two species. In L. albus, the phenolic content and lipid peroxidation were related with the Cu concentration, whereas the Zn and Cd concentrations in N. caerulescens were more closely related to glutathione content and lipid peroxidation. Interestingly, phytochelatins were only found in L. albus grown in polluted soils. Hence, the two species differed with respect to the TEs which provoked stress and the biochemical indicators of the stress, there being a close relationship between the accumulation of TEs and their associated stress indicators in the different plant organs.

Phytostabilisation with Mediterranean shrubs and liming improved soil quality in a pot experiment with a pyrite mine soil.

Phytoremediation can be a suitable option to manage derelict mine soils. A pot experiment was carried out under semi-controlled conditions with a mine-impacted soil. A further contamination event was mimicked by applying 5% of pyritic sludge. Four species were planted in pots (Myrtus communis, Retama sphaerocarpa, Rosmarinus officinalis and Tamarix gallica), and some pots remained unplanted as a control. The substrates were moderately to highly contaminated, mainly with arsenic and zinc. The strong acidification induced by the pyritic sludge was buffered with lime and plants survived in all the pots. Liming provoked an effective immobilisation of metals and arsenic. Plant establishment decreased labile As in the substrate by 50%, mainly M. communis, although the levels of extractable metals were not affected by the plants. R. sphaerocarpa and M. communis increased the levels of C and N in the soil by 23% and 34% respectively, and also enhanced enzymatic activities and microbial respiration to the double in some cases. The low transfer of trace elements to shoots limited the phytoextraction rate. Our results support the use of phytostabilisation in Mediterranean mine soils and show how plants of R. sphaerocarpa and M. communis may increase soil health and quality during revegetation.

The fate of arsenic in soil-plant systems.

Arsenic is a natural trace element found in the environment. In some cases and places, human activities have increased the soil concentration of As to levels that exceed hazard thresholds. Amongst the main contributing sources of As contamination of soil and water are the following: geologic origin, pyriticmining, agriculture, and coal burning. Arsenic speciation in soils occurs and is relatively complex. Soils contain both organic and inorganic arsenic species. Inorganic As species include arsenite and arsenate, which are the most abundant forms found in the environment. The majority of As in aerated soils exists as H2AsO4- (acid soils) or HAsO4²- (neutral species and basic). However, HA3sO3 is the predomiant anaerobic soils, where arsenic availability is higher and As(III) is more weakly retained in the soil matrix than is As(V). The availability of As in soils is usually driven by multiple factors. Among these factors is the presence of Fe-oxides and/or phosphorus, (co)precipitation in salts, pH, organic matter, clay content, rainfall amount, etc. The available and most labile As fraction can potentially be taken up by plant roots, although the concentration of this fraction is usually low. Arsenic has no known biological function in plants. Once inside root cells, As(V) is quickly reduced to As(III), and, in many plant species, becomes complexed. Phosphorus nutrition influences As(V) uptake and toxicity in plants, whilst silicon has similar influences on As(III). Plants cope with As contamination in their tissues by possessing detoxification mechanisms. Such mechanisms include complexation and compartmentalization. However, once these mechanisms are saturated, symptoms of phytotoxicity appear. Phytotoxic effects commonly observed from As exposure includes growth inhibition, chlorophyll degradation, nutrient depletion and oxidative stress. Plants vary in their ability to accumulate and tolerate As (from tolerant hyperaccumulators to sensitive excluders), and some plants are useful for soil reclamation and in sustainable agriculture, The status of current scientific knowledge allows us to manage As contamination in the soil-plant system and to mitigate arsenic's effects. Phytoremediation is an emerging technology suitable for reclaiming As-contaminated soils and waters. Phytoextraction has been used to clean As-contaminated soils, although its applicability has not yet reached maturity. Phytostabilization has been employed to reduce environmental risk by confining As as an inert form in soils and has shown success in both laboratory experiments and in field trials. Phytofiltration has been used to treat As-enriched waters. Such treatment removes As when it is accumulated in plants grown in or on water. In agricultural food production, appropriate soil management and plant variety/species selection can minimize As-associated human dis- eases and the transfer of As within the food chain. Selecting suitable plants for use on As-contaminated soils may also enhance alternative land use, such as for energy or raw material production.

Natural attenuation of residual heavy metal contamination in soils affected by the Aznalcóllar mine spill, SW Spain.

Non-amended soils affected by pyritic sludge residues were monitored for 7 years to assess the long-term natural attenuation ability of these soils. The decrease in both the total concentration of elements (particularly As) and (NH(4))(2)SO(4)-extractable fractions of Mn, and Zn, below the maximum permissible levels indicate a successful natural ability to attenuate soil pollution. Soil acidification by pyrite oxidation and rainfall-enhanced leaching were the largest contributors to the reduction of metals of high (Mn, Cu, Zn and Cd) and low (Fe, Al, and As) availability. Periodic use of correlation and spatial distribution analysis was useful in monitoring elemental dispersion and soil property/element relationships.

Using Mediterranean shrubs for the phytoremediation of a soil impacted by pyritic wastes in Southern Spain: a field experiment.

Re-vegetation is the main aim of ecological restoration projects, and in Mediterranean environments native plants are desirable to achieve successful restoration. In 1998, the burst of a tailings dam flooded the Guadiamar river valley downstream from Aznalcóllar (Southern Spain) with sludges that contained elevated concentrations of metals and metalloids, polluting soils and waters. A phytoremediation experiment to assess the potential use of native shrub species for the restoration of soils affected by the spillage was performed from 2005 to 2007, with soils divided into two groups: pH < 5 and pH > 5. Four native shrubs (Myrtus communis, Retama sphaerocarpa, Rosmarinus officinalis and Tamarix gallica) were planted and left to grow without intervention. Trace element concentrations in soils and plants, their extractability in soils, transfer factors and plant survival were used to identify the most-interesting species for phytoremediation. Total As was higher in soils with pH < 5. Ammonium sulphate-extractable zinc, copper, cadmium and aluminium concentrations were higher in very-acid soils, but arsenic was extracted more efficiently when soil pH was >5. Unlike As, which was either fixed by Fe oxides or retained as sulphide, the extractable metals showed significant relationships with the corresponding total soil metal concentration and inverse relationships with soil pH. T. gallica, R. officinalis and R. sphaerocarpa survived better in soils with pH > 5, while M. communis had better survival at pH < 5. R. sphaerocarpa showed the highest survival (30%) in all soils. Trace element transfer from soil to harvestable parts was low for all species and elements, and some species may have been able to decrease trace element availability in the soil. Our results suggest that R. sphaerocarpa is an adequate plant species for phytostabilising these soils, although more research is needed to address the self-sustainability of this remediation technique and the associated environmental changes.

Screening risk assessment tools for assessing the environmental impact in an abandoned pyritic mine in Spain.

This paper describes a new methodology for assessing site-specific environmental impact of contaminants. The proposed method integrates traditional risk assessment approaches with real and variable environmental characteristics at a local scale. Environmental impact on selected receptors was classified for each environmental compartment into 5 categories derived from the whole (chronic and acute) risk assessment using 8 risk levels. Risk levels were established according to three hazard quotients (HQs) which represented the ratio of exposure to acute and chronic toxicity values. This tool allowed integrating in only one impact category all the elements involved in the standard risk assessment. The methodology was applied to an abandoned metal mine in Spain, where high levels of As, Cd, Zn and Cu were detected. Risk affecting potential receptors such as aquatic and soil organisms and terrestrial vertebrates were assessed. Whole results showed that impact to the ecosystem is likely high and further investigation or remedial actions are necessary. Some proposals to refine the risk assessment for a more realistic diagnostic are included.

Efficiency of white lupin in the removal of mercury from contaminated soils: soil and hydroponic experiments.

This study examined the ability of the white lupin to remove mercury (Hg) from a hydroponic system (Hg concentrations 0, 1.25, 2.5, 5 and 10 micromol/L) and from soil in pots and lysimeters (total Hg concentration (19.2 +/- 1.9) mg/kg availability 0.07%, and (28.9 +/- 0.4) mg/kg availability 0.09%, respectively), and investigated the accumulation and distribution of Hg in different parts of the plant. White lupin roots efficiently took up Hg, but its translocation to the harvestable parts of the plant was low. The Hg concentration in the seeds posed no risk to human health according to the recommendations of the World Health Organization, but the shoots should not be used as fodder for livestock, at least when unmixed with other fodder crops. The accumulation of Hg in the hydroponically-grown plants was linear over the concentration range tested. The amount of Hg retained in the roots, relative to the shoots, was almost constant irrespective of Hg dose (90%). In the soil experiments, Hg accumulation increased with exposure time and was the greater in the lysimeter than in the pot experiments. Although Hg removal was the greater in the hydroponic system, revealing the potential of the white lupin to extract Hg, bioaccumulation was the greatest in the lysimeter-grown plants; the latter system more likely reflects the true behaviour of white lupin in the field when Hg availability is a factor that limits Hg removal. The present results suggest that the white lupin could be used in long-term soil reclamation strategies that include the goal of profitable land use in Hg-polluted areas.

Feasibility of arsenic phytostabilisation using Mediterranean shrubs: impact of root mineralisation on As availability in soils.

An incubation experiment was carried out to evaluate the rate of arsenic release from roots to soil during root mineralisation, in order to determine the viability of phytostabilisation of metal-polluted soil. Arsenic (As)-loaded roots were mixed with the soil, monitoring arsenic, phosphorus, copper and zinc solubility and pH. Arsenic dynamics in soil during root mineralisation depend on various factors such as mineralisation rate, pH and soil chemistry. After 56 days incubation, only the soil with T. gallica roots showed higher NH4(SO4)2-extractable As than soil without roots. Phosphorus release was positively correlated with extractable arsenic. Extractable Cu depended on root mineralisation but Zn solubility was controlled by soil pH. On the whole, species with high As accumulation in roots and a high mineralisation rate may release arsenic in soluble forms in the soil immediately after root addition, but the soil can partly retain arsenic subsequently. In this respect, the risk of As mobilisation due to root decomposition was found to be low, confirming the prospects for long-term phytomanagement and phytostabilisation of arsenic.

Heavy metals distribution in soils surrounding an abandoned mine in NW Madrid (Spain) and their transference to wild flora.

The present work concerns the distribution and mobility of heavy metals (Fe, Mn, Cu, Zn and Cd) in the surrounding soils of a mine site and their transfer to wild flora. Thus, soils and plants were sampled from a mining valley in NW Madrid (Spain), and total and extractable heavy metals were analysed. Soils affected by mining activities presented total Cd, Cu and Zn concentrations above toxic thresholds. The percentage of extractable element was highest for Cd and lowest for Cu. A highly significant correlation was observed between the total and extractable concentrations of metals in soils, indicating that, among the factors studied, total metals concentration is the most relevant for heavy metals extractability in these soils. (NH(4))(2)SO(4)-extractable metal concentrations in soils are correlated better with metal concentrations in several plant species than total metals in soils, and thus can be used as a suitable and robust method for the estimation of the phytoavailable fraction present in soils. Twenty-five vascular plant species (3 ferns and 22 flowering plants) were analysed, in order to identify exceptional characteristics that would be interesting for soil phytoremediation and/or reclamation. High Cd and Zn concentrations have been found in the aerial parts of Hypericum perforatum (Cd), Salix atrocinerea (Cd, Zn) and Digitalis thapsi (Cd, Zn). The present paper is, to the best of our knowledge, the first report of the metal accumulation ability of the two latter plant species. The phytoremediation ability of S. atrocinerea for Cd and Zn was estimated, obtaining intervals of time that could be considered suitable for the phytoextraction of polluted soils.

Evolution of arsenate toxicity in nodulated white lupine in a long-term culture.

White lupine is an As-resistant legume that is of interest for phytoremediation of As-contaminated soils. To achieve successful phytoremediation, monitoring of the nutritional status of the selected plant species during the entire culture cycle is required to maintain a plant cover with high biomass production. A long-term pot experiment was carried out with nodulated lupine grown on perlite with 10 and 100 microM As concentrations. The reproductive period (from 10 weeks) was the most sensitive phenologic stage of white lupine to long-term As exposure. The 10 microM As treatment increased the uptake and translocation of micronutrients, except for Cu, mainly at flowering with As levels in pods below the statutory limit (1 mg kg (-1) fresh weight). However, the 100 microM As treatment induced significant differences compared to the control. These findings confirm the relatively high resistance of white lupine to arsenate and support the use of this species in phytoremediation and/or revegetation of As-contaminated sites, with special attention on P and Cu nutrition at flowering.

Comparison of arsenic resistance in Mediterranean woody shrubs used in restoration activities.

Myrtus communis, Arbutus unedo and Retama sphaerocarpa are Mediterranean shrubs widely used in revegetation of semiarid degraded soils. The aim of this work is to study the resistance of these plants to arsenic under controlled conditions, in order to evaluate their potential use in revegetation and/or phytoremediation of As-polluted soils. R. sphaerocarpa showed higher resistance to As than M. communis or A. unedo according to its higher EC50, P status and P/As molar ratio in both, roots and shoots, and the lower increases in lipid peroxidation and decrease of chlorophyll levels in response to arsenic, while the highest arsenate sensitivity was obtained for A. unedo. Arsenic was mainly retained in roots, and, although M. communis accumulated higher arsenic amounts than the other two species, R. sphaerocarpa showed the highest root to shoot transfer. Most of the studied parameters (chlorophylls, MDA and total thiols) showed significant correlation with arsenic concentration in roots and leaves of plants, so they can be useful indexes in the diagnosis of arsenic toxicity in these species. According to our results, both M. communis and R. sphaerocarpa could be used in the revegetation of moderately arsenic contaminated sites.

Chelate-assisted phytoextraction of heavy metals in a soil contaminated with a pyritic sludge.

The occurrence of many polluted areas as that affected by the accident of the Aznalcóllar pyrite mine has promoted phytoremediation as a technology able to reduce the risk of heavy metal contamination at low cost. White lupin plant has been considered a good candidate for phytoremediation. We studied the capacity of several complexing agents to improve the ability of white lupin for heavy metal phytoremediation in soils with multi-elemental pollution from acid pyritic sludge. Solution-soil interaction was studied and pot experiments with sludge-affected soil were carried out to this end. The interaction experiments indicated that EDTA and NTA were more efficient than malate and citrate in solubilizing metals (Fe, Mn, Cu, Zn, Cd), with minimum differences between EDTA and NTA. The pot trial showed that NTA was able to mobilize toxic elements from sludge-polluted soil and hence increasing their concentrations in plant (Mn, Cu, Zn, As, Cd). However, the NTA treatment promoted an increase of toxic elements concentrations, especially for As, Cd, Pb, in the lixiviates exceeding the maximum permissible levels, so a careful management of chelate is necessary.