Abandoned coal mining sites: using ecotoxicological tests to support an industrial organic sludge amendment

The different stages involved in coal mining-related activities result in a degraded landscape and sites associated with large amounts of dumped waste material. Remediation of these contaminated soils can be carried out by application of industrial organic sludge if the concerns regarding the potential negative environmental impacts of this experimental practice are properly addressed. In this context, the objective of this study was to use ecotoxicological tests to determine the quantity of organic industrial sludge that is required as a soil amendment to restore soil production while avoiding environmental impact. Chemical analysis of the solids (industrial sludge and soil) and their leachates was carried out as well as a battery of ecotoxicity tests on enzymes (hydrolytic activity), bacteria, algae, daphnids, earthworms, and higher plants, according to standardized methodologies. Solid and leachate samples of coal-contaminated soil were more toxic than those of industrial sludge towards enzyme activity, bacteria, algae, daphnids, and earthworms. In the case of the higher plants (lettuce, corn, wild cabbage, and Surinam cherry) the industrial sludge was more toxic than the coal-contaminated soil, and a soil/sludge mixture (66:34 % dry weight basis) had a stimulatory effect on the Surinam cherry biomass. The ecotoxicological assessment of the coal-contaminated soil remediation using sludge as an amendment is very important to determine application rates that could promote a stimulatory effect on agronomic species without negatively affecting the environment.     

Ecotoxicological assessment of metal-polluted urban soils using bioassays with three soil invertebrates

This study aimed at assessing the quality of urban soils by integrating chemical and ecotoxicological approaches. Soils from five sites in downtown Naples, Italy, were sampled and characterized for physical-chemical properties and total and water-extractable metal concentrations. Bioassays with Eisenia andrei, Enchytraeus crypticus and Folsomia candida were performed to assess toxicity of the soils, using survival, reproduction and growth as the endpoints. Metal bioaccumulation in the animals was also measured. The properties and metal concentrations of the soils strongly differed. Metal bioaccumulation was related with total metal concentrations in soil and was highest in E. crypticus, which was more sensitive than E. andrei and F. candida. Responses of the three species to the investigated soils seemed due to both metal contamination and soil properties.     

The water-soluble fraction of potentially toxic elements in contaminated soils: relationships between ecotoxicity, solubility and geochemical reactivity

To better understand the impacts posed by soil contamination to aquatic ecosystems it is crucial to characterise the links between ecotoxicity, chemical availability and geochemical reactivity of potentially toxic elements (PTE’s) in soils. We evaluated the adverse effects of water extracts obtained from soils contaminated by chemical industry and mining, using a test battery including organisms from different trophic levels (bacteria, algae and daphnids). These tests provided a quick assessment of the ecotoxicity of soils with respect to possible adverse effects on aquatic organisms although the ecotoxicological responses could be related to the solubility of PTE’s only to a limited extent.The analysis of results of bioassays together with the chemical characterisation of water extracts provided additional relevant insight into the role of conductivity, pH, Al, Fe, and Mn of soil extracts on toxicity to organisms. Furthermore, an important conclusion of this study was that the toxicity of extracts to the aquatic organisms could also be related to the soil properties (pH, Org C and Fe_ox) and to the reactivity of PTE’s in soils which in fact control the soluble fraction of the contaminants.The combined assessment of ecotoxicity in water fractions, solubility and geochemical reactivity of PTE’s in soils provided a more comprehensive understanding of the bioavailability of inorganic contaminants than ecotoxicological or chemical studies alone and can therefore be most useful for environmental risks assessment of contaminated soils.     

Effects of the antibiotics oxytetracycline and tylosin on soil fauna

Antibiotics may enter the terrestrial environment when amending soils with manure. A Note of Guidance on ecological risk assessment of veterinary medicines was issued in January 1998. Hardly any information about ecotoxicological effects of already existing substances are available. This study has tested the effects of two widely used antibiotics, tylosin and oxytetracycline, on three species of soil fauna: Earthworms, springtails and enchytraeids. Neither of the substances had any effect at environmentally relevant concentrations. The lowest observed effect concentration was 3000 mg kg-1 and in many cases no effect was seen even at the highest test concentration of 5000 mg kg-1.     

Leachates from solid wastes: chemical and eco(geno)toxicological differences between leachates obtained from fresh and stabilized industrial organic sludge

The chemical and ecotoxicological characteristics of fresh and stabilized industrial organic sludge leachates were compared to obtain information regarding how the stabilization process can influence the ecotoxic potential of this industrial waste, which could be used for the amendment of degraded soil. Physicochemical analysis of the sludge leachates, as well as a battery of eco(geno)toxicity tests on bacteria, algae, daphnids, and higher plants (including Vicia faba genotoxicity test) and the determination of hydrolytic enzyme activity, was performed according to standard methods. The chemical comparison of the two types of leachate showed that the samples obtained from stabilized sludge had a lower organic content and higher metal content than leachates of the fresh sludge. The eco(geno)toxicological results obtained with aquatic organisms showed that the stabilized sludge leachate was more toxic than the fresh sludge leachate, both originating from the same industrial organic sludge sample. Nevertheless, phytotoxicity tests carried out with a reference peat soil irrigated with stabilized sludge leachate showed the same toxicity as the fresh sludge leachate. In the case of the industrial solid organic sludge studied, stabilization through a biodegradation process promoted a higher metal mobility/bioavailability/eco(geno)toxicity in the stabilized sludge leachate compared to the fresh sludge leachate.     

Derivation of soil values for the path ‘Soil-Soil Organisms’ for metals and selected organic compounds using species sensitivity distributions

BACKGROUND, AIMS AND SCOPE: According to the German Federal Soil Protection Act, the natural function of soil as a habitat for human beings, animals, plants and soil organisms is, among other things, to be protected by deriving soil values for important chemicals regarding their amounts in the environment, their persistence and/or their toxicity. This contribution presents the results of the mathematical derivation of such values for nine metals and ten organic substances from soil ecotoxicological effect values available in the literature for microbial processes, plants and soil invertebrates. MATERIAL AND METHODS: Ecotoxicological data were mostly extracted from published papers and reports and had to originate from valid studies that were performed according to internationally standardised guidelines (e.g. ISO) or were otherwise well documented, plausible and performed according to accepted laboratory practice. As test results, both structural (i.e., effects on mortality, growth or reproduction) and functional (i.e., effects on microbial activity or organic matter breakdown) parameters were included. The derivation of soil values was performed using the distribution based extrapolation model (DIBAEX) and EC(50)s (Effective Concentration) as input data. RESULTS: For 19 compounds, soil values could be calculated. In 18 of these 19 cases clear laboratory ecotoxicological effects (i.e., EC50 values) below the calculated soil value have been found in the literature. DISCUSSION: In those few cases where a comparison with field studies is possible, effects have been observed in the same order of magnitude as the calculated soil values. A comparison with other similar approaches confirmed the plausibility of the calculated values. CONCLUSIONS: The DIBAEX-method is a feasible and widely accepted method for deriving soil values from ecotoxicological input data. Data availability was already satisfactory for some substances, but other substances, especially organics, were only poorly covered. The soil values presented here were based on EC50 input data. However, depending on the protection level aimed at by using soil values in legislation, it might be appropriate to use other input data such as NOECs in the derivation process. RECOMMENDATIONS AND PERSPECTIVES: It is recommended to generate an appropriate number of data for further relevant substances by means of a test battery or multi-species approaches such as terrestrial model ecosystems. These tests should also consider the influence of the bioavailability of substances. A final recommendation for legally binding soil values demands a plausibility check of the mathematically derived values. This should include a comparison with natural background concentrations, soil values for other pathways and soil values used in legislation of other countries. Finally, expert judgement always has to be considered.     

用水生生物对重金属污染土壤进行生态毒理评价

应用斜生栅藻（Scenedesmus obliquus）和明亮发光杆菌T3（Photobacterium phosphoreum）急性毒性实验方法对重金属污染土壤的进行生态毒理评价，结果表明，斜生栅藻的生长繁殖率和发光菌的相对发光度与土壤中的重金属含量明显相关，并且随重金属投加量的增加，其生长繁殖率和发光度逐渐降低。对3种测试参数进行比较可知，斜生栅藻细胞数增长率是最敏感的土壤毒性检测指标。土壤的毒性在复合污染的条件下比在单一污染的条件下高很多。本研究的结果可以为污染土壤的优先修复提供理论依据。     

Toxicity of emerging energetic soil contaminant CL-20 to potworm Enchytraeus crypticus in freshly amended or weathered and aged treatments

We investigated the toxicity of an emerging polynitramine energetic material hexanitrohexaazaisowurtzitane (CL-20) to the soil invertebrate species Enchytraeus crypticus by adapting then using the Enchytraeid Reproduction Test (ISO/16387:2003). Studies were designed to develop ecotoxicological benchmark values for ecological risk assessment of the potential impacts of accidental release of this compound into the environment. Tests were conducted in Sassafras Sandy Loam soil, which supports relatively high bioavailability of CL-20. Weathering and aging procedures for CL-20 amended into test soil were incorporated into the study design to produce toxicity data that better reflect soil exposure conditions in the field compared with the toxicity in freshly amended soils. Concentration-response relationships for measurement endpoints were determined using nonlinear regressions. Definitive tests showed that toxicities for E. crypticus adult survival and juvenile production were significantly increased in weathered and aged soil treatments compared with toxicity in freshly amended soil, based on 95% confidence intervals. The median effect concentration (EC50) and EC20 values for juvenile production were 0.3 and 0.1 mg kg-1, respectively, for CL-20 freshly amended into soil, and 0.1 and 0.035 mg kg-1, respectively, for weathered and aged CL-20 soil treatments. These findings of increased toxicity to E. crypticus in weathered and aged CL-20 soil treatments compared with exposures in freshly amended soils show that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information on ecotoxicological effects of emerging energetic contaminants in soil.     

Mercury in United Kingdom topsoils; concentrations, pools, and Critical Limit exceedances

The median total mercury concentration in 898 UK rural topsoils, sampled between 1998 and 2008, was 0.095 μg g⁻¹. Approximate adjustment for unreactive metal produced an estimate of 0.052 μg g⁻¹ for reactive Hg. The highest concentrations were in the north and west, where organic-rich soils with low bulk densities dominate, but the spatial pattern was quite different if soil Hg pools (mg m⁻²) were considered, the highest values being near to the industrial north of England and London. Possible toxic effects of Hg were best evaluated by comparison with soil Critical Limits expressed as ratios of Hg to soil organic matter, or soil solution Hg²⁺ concentrations, estimated by chemical speciation modelling. Only a few percent of the rural UK soils showed exceedance, and this also applied to rural soils from the whole of Europe. UK urban and industrial soils had higher Hg concentrations and more cases of exceedance.     

用水生生物对重金属污染土壤进行生态毒理评价

应用斜生栅藻(Scenedesmusobliquus)和明亮发光杆菌T3(Photobacteriumphosphoreum)急性毒性实验方法对重金属污染土壤的进行生态毒理评价,结果表明,斜生栅藻的生长繁殖率和发光菌的相对发光度与土壤中的重金属含量明显相关,并且随重金属投加量的增加,其生长繁殖率和发光度逐渐降低。对3种测试参数进行比较可知,斜生栅藻细胞数增长率是最敏感的土壤毒性检测指标。土壤的毒性在复合污染的条件下比在单一污染的条件下高很多。本研究的结果可以为污染土壤的优先修复提供理论依据。     

The application of bioassays as indicators of petroleum-contaminated soil remediation

Bioremediation has proven successful in numerous applications to petroleum contaminated soils. However, questions remain as to the efficiency of bioremediation in lowering long-term soil toxicity. In the present study, the bioassays Spirotox, Microtox, Ostracodtoxkit F, umu-test with S-9 activation, and plant assays were applied, and compared to evaluate bioremediation processes in heavily petroleum contaminated soils. Six higher plant species (Secale cereale L., Lactuca sativa L., Zea mays L., Lepidium sativum L., Triticum vulgare L., Brassica oleracea L.) were used for bioassay tests based on seed germination and root elongation. The ecotoxicological analyses were made in DMSO/H2O and DCM/DMSO soil extracts. Soils were tested from two biopiles at the Czechowice oil refinery, Poland, that have been subjected to different bioremediation applications. In biopile 1 the active or engineered bioremediation process lasted four years, while biopile 2 was treated passively or non-engineered for eight months. The test species demonstrated varying sensitivity to soils from both biopiles. The effects on test organisms exposed to biopile 2 soils were several times higher compared to those in biopile 1 soils, which correlated with the soil contaminants concentration. Soil hydrocarbon concentrations indeed decreased an average of 81% in biopile 1, whereas in biopile 2 TPH/TPOC concentrations only decreased by 30% after eight months of bioremediation. The bioassays were presented to be sensitive indicators of soil quality and can be used to evaluate the quality of bioremediated soil. The study encourages the need to combine the bioassays with chemical monitoring for evaluation of the bioremediation effectiveness and assessing of the contaminated/remediated soils.     

Comparison of solid and liquid-phase bioassays using ecoscores to assess contaminated soils

Bioassays on aqueous and solid phases of contaminated soils were compared, belonging to a wide array of trophic and response levels and using ecoscores for evaluating ecotoxicological and genotoxicological endpoints. The method was applied to four coke factory soils contaminated mainly with PAHs, but also to a lesser extent by heavy metals and cyanides. Aquatic bioassays do not differ from terrestrial bioassays when scaling soils according to toxicity but they are complementary from the viewpoint of ecological relevance. Both aquatic and terrestrial endpoints are strongly correlated with concentrations of 3-ring PAHs. This evaluation procedure allows us to propose a cost-effective battery which embraces a wide array of test organisms and response levels: it includes two rapid bioassays (Microtox(?) and springtail avoidance), a micronucleus test and three bioassays of a longer duration (algal growth, lettuce germination and springtail reproduction). This battery can be recommended for a cost-effective assessment of polluted/remediated soils.     

Non-destructive pollution exposure assessment in the European hedgehog (Erinaceus europaeus): IV. Hair versus soil analysis in exposure and risk assessment of organochlorine compounds

Few ecotoxicological studies on mammals use non-destructive methodologies, despite the growing ethical concern over the use of destructive sampling methods. In the present study we assessed exposure of hedgehogs (Erinaceus europaeus) to polychlorinated biphenyls (PCBs), dichloro-diphenyl-trichloroethanes (DDTs), hexachlorocyclohexanes (HCHs) and hexachlorobenzene (HCB), by investigating concentrations of these compounds in soils and hedgehog hair from seven study sites around the urban area of Antwerp, Belgium. No relationships were observed between organochlorine compound concentrations in soils and hair from the different study areas. Furthermore, the individual variation of contamination levels in hair within study sites was high, especially for HCHs and HCB, and hair and soil had different relative profiles for PCBs, DDTs and HCHs. Our results show that concentrations of organochlorine compounds in soils alone are not predictive of the risk of these pollutants to hedgehogs and that tissue analyses are preferred to soil analyses in exposure and risk assessment studies.     

Polychlorinated biphenyls (PCBs) in soils of the Moscow region: concentrations and small-scale distribution along an urban-rural transect

In soils of the Moscow region, we examined PCB concentrations in bulk samples and aggregate fractions. Topsoils under grassland and forest at five locations along a southeast-bound transect from Moscow and at a northeastern background location (grassland only) were analysed. We collected aggregates >1 cm and fractionated them into interior and exterior portions and also analysed the remaining soil without the aggregates >1 cm. The concentrations of the sum of 17 PCBs (sigma17PCBs) in 35 bulk soil samples ranged from 3.1 to 42 microg kg(-1). This was 48-61% of the sigma33PCBs determined in 23 selected samples. The congeners 138(+158), 101 and 52 were most abundant. All PCB concentrations and the degree of chlorination declined with increasing distance from Moscow. The PCBs were accumulated in the aggregate exterior (on average 146% of the sigma17PCBs in bulk soil). We conclude that the ecotoxicological risk of PCBs in soils may not be properly assessed with the conventional bulk soil analysis.     

Comparison of solid-phase bioassays and ecoscores to evaluate the toxicity of contaminated soils

Five bioassays (inhibition of lettuce germination and growth, earthworm mortality, inhibition of springtail population growth, avoidance by springtails) were compared, using four coke factory soils contaminated by PAHs and trace elements, before and after biotreatment. For each bioassay, several endpoints were combined in an ‘ecoscore’, a measure of test sensitivity. Ecoscores pooled over all tested bioassays revealed that most organisms were highly sensitive to the concentration of 3-ring PAHs. When four soils were combined, behavioural tests using the springtail Folsomia candida showed higher ecoscores, i.e. they were most sensitive to soil contamination. However, despite overall higher sensitivity of behavioural tests, which could be used for cheap and rapid assessment of soil toxicity, especially at low levels of contamination, some test endpoints were more sensitive than others, and this may differ from a soil to another, pointing to the need for a battery of bioassays when more itemized results are expected.     

Effect of pH control at the anode for the electrokinetic removal of phenanthrene from kaolin soil

Polycyclic aromatic hydrocarbon (PAH)-contaminated soils exist at numerous sites, and these sites may threaten public health and the environment because many PAH compounds are toxic, mutagenic, and/or carcinogenic. PAHs are also hydrophobic and persistent, so conventional remediation methods are often costly or inefficient, especially when the contaminants are present in low permeability and/or organic soils. An innovative technique, electrokinetically enhanced in situ flushing, has the potential to increase soil-solution-contaminant interaction and PAH removal efficiency for low permeability soils; however, the electrolysis reaction at the anode may adversely affect the remediation of low acid buffering capacity soils, such as kaolin. Therefore, the objective of this study was to improve the remediation of low acid buffering soils by controlling the pH at the anode to counteract the electrolysis reaction. Six bench-scale electrokinetic experiments were conducted, where each test employed one of three different flushing solutions, deionized water, a surfactant, or a cosolvent. For each of these solutions, tests were performed with and without a 0.01 M NaOH solution at the anode to control the pH. The test using deionized water with pH control generated a higher electroosmotic flow than the equivalent test performed without pH control, but the electroosmotic flow difference between the surfactant and cosolvent tests with and without pH control was minor compared to that observed with the deionized water tests. Controlling the pH was beneficial for increasing contaminant solubilization and migration from the soil region adjacent to the anode, but the high contaminant concentrations that resulted in the middle or cathode soil regions indicates that subsequent changes in the soil and/or solution chemistry caused contaminant deposition and low overall contaminant removal efficiency.     

Fate and behaviour of phenanthrene in the natural and artificial soils

For the assessment of contaminated soils ecotoxicological tests are used to estimate the bioavailability of contaminants in soil samples. Terrestrial tests reveal the habitat function of soils, and parameters applied in tests involving microorganisms include respiration activity and potential ammonium oxidation. For such tests, the threshold values needed to assess the results have already been established in guidelines ISO 17155 and ISO 15685. In this paper, we discuss about the respiration activity and potential ammonium oxidation results obtained from a wide variety of soils with different physico-chemical properties and levels of contamination. These results show that microbial respiration and potential ammonium oxidation have different sensitivities to various classes of contaminants. We demonstrated that both organic and inorganic contaminants influence potential ammonium oxidation, whereas microbial respiration is predominantly affected by biodegradable organic contaminants. These differences might be useful for more detailed assessments of soil contamination, leading to different recommended actions depending on which parameter is affected.     

Mobility of metals and metalloids in a multi-element contaminated soil 20 years after cessation of the pollution source activity

Knowledge of trace element concentrations and mobility is important in the ecotoxicological assessment of contaminated soils. We analysed soil pore water under field conditions to provide new insights into the mobility of residual contaminants in the surface 50cm of a highly contaminated woodland soil. Cadmium and Zn were highly mobile in the acidic soil, concentrations increasing with depth in soil pore water, showing considerable downward mobility. High levels of surface organic matter restricted the solubility of Cu, Pb and Sb, with highest concentrations being found close to the surface. Dissolved organic carbon in pore water had a strong influence on mobility of Cu, Zn, Pb and Sb. Elevated As had moved from the organic surface horizons but was largely immobilised in deeper layers and associated with Fe and Al oxides. The measured differential mobility of pollutants in the present study is highly relevant to protection of groundwater and other receptors.     

Ecotoxicological evaluation of in situ bioremediation of soils contaminated by the explosive 2,4,6-trinitrotoluene (TNT)

To evaluate the environmental relevance of in situ bioremediation of contaminated soils, effective and reliable monitoring approaches are of special importance. The presented study was conducted as part of a research project investigating in situ bioremediation of topsoils contaminated by the explosive 2,4,6-trinitrotoluene (TNT). Changes in soil toxicity within different experimental fields at a former ordnance factory were evaluated using a battery of five bioassays (plant growth, Collembola reproduction, soil respiration, luminescent bacteria acute toxicity and mutagenicity test) in combination to chemical contaminant analysis. Resulting data reveal clear differences in sensitivities between methods with the luminescent bacteria assay performed with soil leachates as most sensitive toxicity indicator. Complete test battery results are presented in so-called soil toxicity profiles to visualise and facilitate the interpretation of data. Both biological and chemical monitoring results indicate a reduction of soil toxicity within 17 months of remediation.     

污染土壤修复效果评定方法的研究

在实施污染土壤修复的环境工程后，需要通过灵敏和有效的评定方法对污染土壤修复的效果进行评定。然而，单纯依靠化学方法进行污染土壤修复效果的评定，不能揭示土壤的整体质量特征，因此需要生态毒理方法作为相互补充的手段。本文概述了植物毒性评定法、陆生无脊椎动物评定法和土壤微生物评定法及其在污染土壤修复效果评定中的应用，并对污染土壤修复效果评定方法的发展前景进行展望。