The influence of soil properties on the toxicity of molybdenum to three species of soil invertebrates

Mo toxicity to earthworms (Eisenia andrei), Collembola (Folsomia candida) and enchytraeids (Enchytraeus crypticus) was determined in 10 European soils and a standard artificial soil, freshly spiked with Na₂MoO₄, after 28 days exposure. Mo affected survival only in three low pH sandy soils; in all other soils LC50 was >3200 mg Mo/kg dry soil. EC50 values for the reproduction toxicity of Mo were 129-2378 mg/kg for earthworms, 72->3396 mg/kg for Collembola, and 301->2820 mg/kg for enchytraeids. Variation in toxicity among soils could not be explained by differences in available (pore water, water and 0.01 M CaCl₂ extractable) Mo concentrations. Clay content best predicted the EC50 for Mo toxicity to earthworms, while toxicity of Mo for enchytraeids was best described by soil pH. For Collembola no relationships could be derived due to the absence of toxicity in most soils. Soil properties had a strong but species-specific effect on Mo toxicity to soil invertebrates.     

Bioavailability of Copper and Zinc in Mining Soils

The soil-contact exposure pathway can be the main driver of ecological risk assessments. There is currently no standard method to measure bioavailability of metals in soil to ecological receptors, yet the influence of metal bioavailability on toxicity has been known for decades and is a major factor influencing risk to ecological receptors. Bioavailability is to a large degree governed by varying soil characteristics within and among sites, yet ecological screening benchmarks are often derived on a total-concentration basis. We compared a calcium chloride (CaCl₂) extraction, cyclodextrin extraction, simulated earthworm gut (SEG) test, earthworm kinetic bioaccumulation test, and metal residues in plant tissues with a battery of invertebrate and toxicity tests using mining soils consisting of high organic-matter content cocontaminated with copper (Cu) and zinc (Zn). Earthworm (Eisenia andrei) tissue concentrations of Cu and Zn were regulated and were not predictive of invertebrate toxicity. All chemical measures of bioavailability correlated with several biological responses; however, CaCl₂-extractable Cu and SEG-extractable Cu and Zn best predicted effects to E. andrei. Total Cu concentrations in soil best correlated with effects to plants. Overall, a chemical measure was the best predictor of toxicity to each organism compared with biological measures, although the exact measure was dependent on organism and end point. Chemical-extraction techniques provide relatively quick, inexpensive indicators of essential metal bioavailability compared with biological measures; however, no single measure was indicative of all effects to all organisms.     

Ecotoxicity of Aged Uranium in Soil Using Plant,Earthworm and Microarthropod Toxicity Tests

Discrepancies about probable no effect concentrations (PNEC) for uranium in soils may be because toxicity tests used freshly contaminated soils. This study used 3 soils amended with a range of uranium concentrations 10 years previously. The toxicity tests with northern wheatgrass (Elymus lanceolatus); earthworm (Eisenia andrei) were not affected below ~1,000 mg U kg⁻¹, and the soil arthropod Folsomia candida was not affected below ~350 mg U kg⁻¹. Survival of Orthonychiurus folsomi was diminished 20% (EC₂₀) by ~85–130 mg U kg⁻¹, supporting a PNEC in the range of 100–250 mg U kg⁻¹ as derived previously.     

Effect of leaching and aging on the bioavailability of lead to the springtail Folsomia candida

Because it is unclear if leaching can account for differences in metal bioavailability observed between metal-spiked soils and historically contaminated field soils, we simultaneously assessed Pb toxicity to the springtail Folsomia candida in three transects of Pb-contaminated soils and in leached and unleached soils spiked at similar total Pb concentrations. Total Pb concentrations of 3,877 mg/kg dry weight and higher always caused significant effects on F. candida reproduction in the spiked soils. In the transects, only the soil with the highest Pb concentration of 14,436 mg/kg dry weight significantly affected reproduction. When expressed as pore-water concentrations, reproduction was never significantly affected at Pb concentrations of 0.539 mg/L, whereas reproduction was always significantly affected at Pb concentrations of 0.678 mg/L and higher, independent of the soil treatment. These results indicate that pore-water Pb concentrations can explain, at least in part, the observed differences in the toxicity data expressed as total Pb concentrations. Leaching after the spiking procedure only caused small differences in Pb toxicity and, therefore, cannot account for toxicity differences between laboratory-spiked soils and historically contaminated field soils.     

Uptake and Elimination of Cadmium and Zinc by Eisenia andrei During Exposure to Low Concentrations in Artificial Soil

Bioaccumulation factors (BAFs) are often used in the risk assessment process to estimate trophic transfer of contaminants such as metals from soil. BAFs can be derived from laboratory studies through the determination of steady-state concentrations or kinetic estimation methods. In this study, bioaccumulation tests were performed with artificial soil spiked at low levels with cadmium or zinc to determine uptake and elimination kinetics of both metals by the compost worm Eisenia andrei. The metal-amended soils were acclimated for 21 days prior to the test, after which worms were individually incubated in the soils. The uptake phase comprised 0–21 days, after which the test organisms were transferred into clean soil and the elimination phase continued for an additional 21 days. Subsamples of soil and earthworms (whole body) were collected from independent replicates throughout the uptake phase and elimination phase and analyzed for total metal concentrations. Uptake of Cd in E. andrei increased linearly with time and did not reach steady state within the testing period. Cd uptake and excretion were described by a one-compartment first-order kinetics model. Zn concentrations rapidly increased in E. andrei after 1 day of exposure but subsequently decreased to background levels throughout the remainder of the uptake phase; internal Zn concentrations did not change from background levels during the elimination phase. Kinetic BAFs were calculated for Cd and Zn. Cd is a nonessential metal that is bioaccumulated at a relatively rapid rate, while Zn is an essential metal, and as such, it is regulated by E. andrei. Metal essentiality and concentration significantly impact bioaccumulation of metals by terrestrial invertebrates.     

Potentiation Effect of Metolachlor on Toxicity of Organochlorine and Organophosphate Insecticides in Earthworm Eisenia andrei

Acetylcholinesterase, glutathione-S-transferase and catalase activities were determined in earthworms Eisenia andrei exposed to insecticides (endosulfan, temephos, malathion, pirimiphos-methyl) alone and in a binary combination with the herbicide metolachlor. Metolachlor individually was not acutely toxic, even at high concentrations applied; however, in the treated earthworms metolachlor enhanced the toxicity of endosulfan and temephos by significantly reducing the acetylcholinesterase activity. In binary combination with malathion and pirimiphos-methyl, metolachlor did not increase toxicity. The potentiation character of metolachlor is specific rather than general, and probably depends on the chemical structure of pesticides in the mixture.     

Continuous Monitoring of Folsomia candida (Insecta: Collembola) in a Metal Exposure Test

Current recommended ecotoxicological tests with the parthenogenetic springtail Folsomia candida using standard OECD soil do not allow for continuous monitoring during the exposure period. Effects of chemicals cannot be determined until the end of the experiment (typically after 4 weeks), since the animals stay below the soil surface. In this study, F. candida were maintained on a plaster of Paris/graphite substrate for 7 weeks and were supplied with an aqueous suspension of yeast contaminated with Cd, Cu, Pb, and Zn as nitrate salts. Growth rate, time to first batch of eggs, quantity of food consumed, and the presence of graphite in the gut (a sign of avoidance of yeast) were all affected by metal contaminated diets. The relative toxicities of Cd:Cu:Pb:Zn in the yeast were 1.0:1.07:12.0:4.3, respectively (on a weight basis) with Cd being the most toxic. Internal body concentrations increased, and the concentration factor (metal concentration in F. candida/metal concentration in yeast) decreased with increasing metal exposure. In general, metals are much less toxic when added to the food of F. candida than when incorporated into soil in standard tests. It is suggested that Collembola have a greater tolerance of metals in the diet since they avoid contaminated food, and are able to excrete assimilated metals at moulting via exfoliation of the midgut epithelium where the elements are retained as part of a storage–detoxification system. The methodology described in this article allows effects on growth to be observed as early as 7 days after the beginning of the experiment.     

Toxicity and Bioaccumulation of Copper to Black Bindweed (Fallopia convolvulus) in Relation to Bioavailability and the Age of Soil Contamination

The use of ecotoxicity test results obtained in the laboratory for prediction of effects of potentially toxic concentrations of chemicals in the field is hampered by several factors differing between the laboratory and the field situations. One important factor is the binding of test chemicals to soil, which is affected by the age of the contamination and soil type. The present study investigated the effect of contamination age by introducing an aging period of 1 to 12 weeks between mixing the test substance, copper sulfate, in with soil and introducing the test plant, Fallopia convolvulus (L.) A. Löve. Copper accumulation, emergence of cotyledons, and growth were assessed and related to total (boiling with HNO₃) and 0.01 M CaCl₂- and DTPA-extractable soil copper concentrations. Aging of the copper-contaminated soil had only small effects on bioaccumulation of copper, copper toxicity, and extractable soil copper fractions. Soil copper had no effect on emergence of cotyledons. Estimated EC₅₀ values for shoot and root growth averaged 280 mg Cu/kg. Effects on growth in these laboratory-treated soils were much more severe than in a study performed in soil from an old copper-contaminated field site. Neither CaCl₂- nor DTPA-extractable copper fractions could explain all of the differences in effects between aged spiked soil and field soil. The accumulation pattern for roots and shoots of F. convolvulus indicated that excessive copper was accumulated and adsorbed mainly by the fine roots, whereby the copper concentrations of other plant parts were kept low until the plant was no longer able to maintain this regulation. An internal threshold for effects on growth of about 20 mg Cu/kg shoot dry weight was estimated, coinciding with a soil copper concentration of approximately 200 mg/kg.     

Ecotoxicological Evaluation of Sewage Sludge Contaminated with Zinc Oxide Nanoparticles

The objective of this work was to evaluate the ecotoxicological qualitative risk associated with the use of sewage sludge containing Zn oxide nanoparticles (ZnO-NPs) as soil amendment. A sludge-untreated soil and two sludge-treated soils were spiked with ZnO-NPs (0–1,000 mg/kg soil). Soil ecotoxicity was assessed with Eisenia fetida (acute and sublethal end points), and the unfilterable and filterable (0.02 μm) soil leachates were tested with a battery of biomarkers using Chlorella vulgaris, Daphnia magna, and the fish cell line RTG-2 (Oncorhynchus mykiss). The production of E. fetida cocoons in sludge-treated soils was lower than that in sludge-untreated soils. The highest effect in the algal growth inhibition test was detected in sludge-untreated soil, most likely caused by the loss of organic matter in these samples. The D. magna results were always negative. Toxic effects (lysosomal cell function and production of reactive oxygen species) in RTG-2 cells were only observed in sludge-treated soils. In general, the toxicity of ZnO-NPs in sludge-treated soils was similar to that of sludge-untreated soil, and the filterable leachate fraction [Zn salt (Zn²⁺)] did not produce greater effects than the unfilterable fraction (ZnO-NPs). Thus, after the addition of ZnO-NP—enriched sewage sludge to agricultural soil, the risk of toxic effects for soil and aquatic organisms was shown to be low. These findings are important because repeated use of organic amendments such as sewage sludge may cause more and more increased concentrations of ZnO-NPs in soils over the long-term.     

Sewage sludge applied to agricultural soil: Ecotoxicological effects on representative soil organisms

Application of sewage sludge to agricultural lands is a current practice in EU. European legislation permits its use when concentrations of metals in soil do not increase above the maximum permissible limits. In order to assess the fate and the effects on representative soil organisms of sewage sludge amendments on agricultural lands, a soil microcosm (multi-species soil system—MS 3) experiment was performed. The MS 3 columns were filled with spiked soil at three different doses: 30, 60 and 120 t ha^?1 fresh wt. Seed plants (Triticum aestivum, Vicia sativa and Brassica rapa) and earthworms (Eisenia fetida) were introduced into the systems. After a 21-d exposure period, a statistically significant increase for Cd, Cu, Zn and Hg concentrations was found for the soils treated with the highest application rate. Dose-related increase was observed for nickel concentrations in leachates. Plants and earthworm metal body burden offer much more information than metal concentrations and help to understand the potential for metal accumulation. Bioaccumulation factor (BAF_plant?soil) presented a different behavior among species and large differences for BAF_{earthworm?soil}, from control or sewage-amended soil, for Cd and Hg were found. B. rapa seed germination was reduced. Statistically significant decrease in fresh biomass was observed for T. aestivum and V. sativa at the highest application rate, whereas B. rapa biomass decreased at any application rate. Enzymatic activities (dehydrogenase and phosphatase) as well as respiration rate on soil microorganisms were enlarged.     

Toxic effects of nine polycyclic aromatic compounds on Enchytraeus crypticus in artificial soil in relation to their properties

The aim of this study was to compare the toxic effects of selected two- and three-ringed PAHs (naphthalene, phenanthrene, and anthracene) and their N-heterocyclic analogs with one (quinoline, acridine, and phenanthridine) or two (quinoxaline, phenazine, and 1,10-phenanthroline) nitrogen atoms on the survival and reproduction of Enchytraeus crypticus in artificial soil. Toxicity of compounds was recalculated to soil pore-water concentrations using the data of chemical analyses of 0.01 M CaCl₂ extracts of spiked soils. When toxicity was based on molar concentrations in pore water (μmol/L), it significantly increased with increasing K_ow value. This relationship indicates nonpolar narcosis as the general toxicity mechanism of the tested compounds. In addition, significant correlation between the toxicity of PACs and their ionization potential has been identified by multidimensional QSAR models.     

Assessing the toxicity of contaminated soils using the nematode Caenorhabditis elegans as test organism

In this study, nine uncontaminated reference soils and 22 contaminated soils with different physico-chemical properties and contamination patterns were tested with a standardized toxicity test, using the nematode, Caenorhabditis elegans, as test organism. Fertility, growth and reproduction of C. elegans in the soils were compared with the exposure in standard soil Lufa St.2.2. C. elegans showed 100% fertility and a very low variability of growth in the reference soils. Although, reproduction varied considerably between the various reference soils, validity criteria (>30 offspring per test organism) were met in all reference soils. Moreover, Lufa St. 2.2 turned out to be a suitable and representative control soil. In order to clearly classify the effects of the polluted soils on C. elegans, toxicity thresholds were derived for nematode fertility (20% inhibition), growth (10% inhibition) and reproduction (40% inhibition) on the basis of the test inherent variability (MDD=minimal detectable difference), as well as their variability between the uncontaminated reference soils (MTI=maximal tolerable inhibition). The contaminated soils showed clear toxic effects on the nematodes, whereas the toxicity was better correlated to organic than to heavy metal contamination in bulk soil. Interestingly, the results of the nematode toxicity test were not well correlated with those of tests with oligochaetes, collembolans and plants, performed with the same soils, showing that the results are not redundant. The toxicity test using C. elegans turned out to be suitable for testing the toxicity of field collected soils and might by a valuable addition to soil test batteries.     

Effect of humic acid during concurrent chronic waterborne exposure of rainbow trout (Oncorhynchus mykiss) to copper, cadmium and zinc

The effects of commercial dissolved organic carbon (DOC) in moderating accumulation, biochemical responses and toxicity of a waterborne mixture of copper (Cu), cadmium (Cd) and zinc (Zn) were investigated during a chronic exposure. Juvenile rainbow trout (Oncorhynchus mykiss) were exposed to a ternary metals mixture containing (nominal concentrations in μg/l): Cu 30, Cd 15, and Zn 150 in hard water (260 mg/l as CaCO₃) with and without addition of 5 mg/l DOC as Aldrich humic acid (HA) for 28 days. Mortality, growth, metals accumulation, ionoregulatory impairment, and oxidative stress response were measured. While growth was unaffected, 19% mortality occurred during the first week of the exposure in fish exposed to the metals mixture without added HA. The early mortality was associated with transitory whole-body sodium (Na) loss and inhibition of branchial Na⁺, K⁺-ATPase activity. Although these ionoregulatory responses mechanistically suggested that Cu was the more potent toxicant than either Cd or Zn, they were not correlated uniquely with elevated tissue Cu concentrations. The effects of HA on accumulation were metal-specific and depended on the organ examined and exposure duration. Specifically, Zn accumulation occurred only in the gill early in the exposure and HA reversed it, while protection against accumulation was absent or complete for Cu and absent or partial for Cd, dependent on tissue and exposure duration. The computed ambient free metal ion activities could explain the Cd but not the Cu and Zn accumulation indicating the involvement of physiological regulatory mechanisms in defining accumulation of essential metals. Surprisingly, the metals mixture (with and without added HA) reduced the concentrations of malondialdehyde (MDA) in gill suggesting induction of reductive rather than oxidative stress. Overall these data indicate that the free metal ion activity alone is not universally a good predictor of metals mixture accumulation and chronic effects nor does consideration of the mechanisms of toxicity unambiguously identify the more potently toxic metal in a mixture.     

Optimization of cow dung spiked pre-consumer processing vegetable waste for vermicomposting using Eisenia fetida

This paper reports the optimization of cow dung (CD) spiked pre-consumer processing vegetable waste (PPVW) for vermicomposting using Eisenia fetida in a laboratory scale study. Vermicomposting process decreased carbon and organic matter concentration and increased N, P and K content in the vermicompost. The C:N ratio was decreased by 45-69% in different vermireactors indicating stabilization of the waste. The heavy metal content was within permissible limits of their application in agricultural soils. It has been concluded from the results that addition of PPVW up to 40% with CD can produce a good quality vermicompost. Whereas, growth and fecundity of E. fetida was best when reared in 20% PPVW+80% CD feed mixture. However, higher percentages of PPVW in different vermireactors significantly affected the growth and fecundity ofworms.     

Acute Toxicity, Toxicokinetics, and Tissue Target of Lead and Uranium in the Clam Corbicula fluminea and the Worm Eisenia fetida: Comparison with the Fish Brachydanio rerio

The general objective of our work was to propose new reference material for chemical toxicity testing and new sentinel organisms for environmental quality survey programs (freshwater or soils). We also wanted to provide basic toxicological data on the environmental effects of uranium. Thus, we conducted a comparative study to establish the acute toxicity and toxicokinetics of lead (Pb) and uranium (U) to the bivalve mollusc Corbicula fluminea and the terrestrial annelid Eisenia fetida andrei and to compare these findings with those of the well-known teleost fish Brachydanio rerio. We then measured the concentration of these metals in various tissues of the clam and the worm after two periods of exposure (4 and 11 days) to identify the affinities of these tissues for Pb and U. Our results have shown that Pb and U are very toxic to Eisenia and relatively nontoxic to Corbicula. By comparison, Pb was relatively nontoxic and U appeared to be very toxic to the fish. The toxicokinetic studies indicated that the three species are able to accumulate Pb and U, the rate and level of accumulation depending both on the species and the metal. We also found that fish and clams depurate the two metals. Data collected for the worm were conflicting: Pb was not depurated whereas tissue concentrations of U declined after the eighth day of exposure. Our study has also shown that the tissue distribution of Pb in the mollusc and in the earthworm differs significantly from that of U, both after 4 and 11 days exposure. In conclusion, these three species showed potential as bioindicators of environmental contamination by metals. Indeed, they could be used in conjunction to test different compartments of an ecosystem: worms for soils, fish for the water column, and clams for the water/sediment interface. Received: 30 October 1996/Accepted: 1 September 1998     

Short-Term Changes of Metal Availability in Soil. Part I: Comparing Sludge-Amended With Metal-Spiked Soils

Sewage sludge application to soils is regulated by its total metal content. However, the real risk of metals is determined by the fraction that is biologically available. The available fraction is highly related to the strength of metal binding by the matrix, which is a dynamic process. The evaluation of the fate of metals in time can contribute increased accuracy of ecological risk assessment. Aiming to evaluate short-term changes in metal availability when metals were applied to soil directly (metal-spiked) or by way of an organic matrix (sludge-amended), a laboratory experiment was performed using open microcosms filled with agricultural soil. A concentration gradient of industrial sludge (11, 15, 55, and 75 t/ha) that was contaminated predominantly with chromium, copper, nickel, and zinc, or soil freshly spiked with the same concentrations of these metals, were applied on top of the agricultural soil. After 0, 3, 6, and 12 weeks, total (HNO(3) 69 %) and 0.01 M CaCl(2)-extractable metal concentrations in soil and metal content in the percolates were measured. Results demonstrated that comparison between sludge-amended and metal-spiked soils may give important information about the role of sludge matrix on metal mobility and availability in soil. In sludge-amended soils, extractable-metal concentrations were independent of the sludge concentration and did not change over time. In metal-spiked soils, metal extractability decreased with time due to ageing and transport of metals to deeper layers. In general, the sludge matrix increased the adsorption of metals, thus decreasing their mobility in soils.     

Utilizing an Earthworm Bioassay (Eisenia andrei) to Assess a South African Soil Screening Value with Regards to Effects from a Copper Manufacturing Industry

Metal contamination of soil due to industrialization has become an increasingly important problem in South Africa. This study aimed to investigate the potential impact of a copper (CuSO₄·5H₂O) production company on the soil environment. Bioassays using Eisenia andrei were performed to assess changes in biomass, reproduction and a biomarker, neutral red retention time, over a 28 day period. Earthworms exposed to soils from the Cu production site differed significantly (p < 0.05) from those exposed to soils 500 m and 5 km away in terms of the measured endpoints. These findings are consistent with the results from the chemical analysis which showed an elevated soil Cu content for both sites closest to the chemical production company compared to the reference site. The results confirm the importance and predictive value of using bioassays in conjunction with chemical analysis during soil quality assessments.     

Usefulness of the sensitivity-resistance index to estimate the toxicity of copper on bacteria in copper-contaminated soils

Examination was made of the fluctuations of numbers of the total bacteria and copper (Cu)-resistant bacteria with soluble/exchangeable Cu (Ex-Cu) fraction in three types of soils spiked with Cu at four concentrations. Drastic increase in Cu-resistant bacteria was observed in three soils spiked with 20 mmol Cu kg(-1) after 2 weeks of incubation, indicating the strong selection of individuals originally resistant to Cu. Adaptation and proliferation of bacteria were also observed in the soil environment under the long-term exposure to extremely high concentration of Cu (800 mg kg(-1) soil of Ex-Cu), deriving from the development of Cu resistance. These bacterial fluctuations and the toxic effects of Cu depended on soil types, due to the chemical forms in which Cu occurs. It was also found that the ratio of Cu-resistant bacterial number to total bacteria was significantly correlated with the amount of Ex-Cu in the soils. This sensitivity-resistance index seems to be useful for evaluating the toxic effects of Cu on the soil bacterial community. Whereas the toxicity of Cu depended on the soil properties, they also changed with time. This phenomenon can be explained by the decrease in the most labile Cu phase, Ex-Cu, with time in the soils.     

Effects of metal-contaminated forest soils from the Canadian shield to terrestrial organisms

The effects of elevated metal concentrations in forest soils on terrestrial organisms were investigated by determining the toxicity of six site soils from northern Ontario and Quebec, Canada, using a battery of terrestrial toxicity tests. Soils were collected from three sites on each of two transects established downwind of nickel (Sudbury, ON, Canada) and copper (Rouyn-Noranda, PQ, Canada) smelting operations. Site soils were diluted to determine if toxicity estimates for the most-contaminated site soils could be quantified as a percent of site soil. Rouyn-Noranda soils were toxic following acute exposure (14 d) to plants, but not to invertebrates (7 d for collembola and 14 d for earthworms). However, Rouyn-Noranda soils were toxic to all species following chronic exposure (21, 35, and 63 d for plants, collembola, and earthworms, respectively). The toxicity of the Rouyn-Noranda site soils did not correspond to the gradient of metal concentrations in soil. Metal-contaminated Sudbury soils were toxic to plants but not to invertebrates, following acute exposure. Chronic exposure to Sudbury soils caused adverse effects to plant growth and invertebrate survival and reproduction. The toxicity of Sudbury soils corresponded to the metal concentration gradient, with one exception: The reference soil collected in October was toxic to collembola following acute and chronic exposure. This study evaluated the applicability of the new Environment Canada terrestrial toxicity test methods, developed using agricultural soils, to forest soils and also provided useful data to assess the ecological risk associated with mixtures of metals in soil.     

Toxicity of four nitrogen-heterocyclic polyaromatic hydrocarbons (NPAHs) to soil organisms

The aims of this study were: (i) to investigate the toxicity of N-heterocyclic polyaromatic hydrocarbons (NPAHs) quinoline, acridine, phenazine, and 1,10-phenanthroline to the soil invertebrates Eisenia fetida, Enchytraeus crypticus, Folsomia candida, and Caenorhabditis elegans, (ii) to compare the toxicity of four NPAHs and the species sensitivity, and (iii) to discuss possible risks of these compounds in soils. Different toxicities were found for the tested NPAHs which might be partially explained by their structure and properties. Effect concentrations expressed as soil pore-water concentrations were related to log K_ow, which indicated narcosis as the most probable mode of toxic action. The species sensitivity decreased in the rank: springtails >enchytraeids=earthworms> nematodes. Predicted no-effect concentration (PNEC) values were calculated for all tested species giving values from 0.5 to 6.8 mg/kg. It is unlikely that there is a risk for soil organisms in natural soils where lower NPAHs concentrations are expected.