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Uptake and toxicity of Ni, Co and their mixtures in Enchytraeus crypticus after different exposure times (4, 7, 10 and 14d) were predicted using the WHAM-FTOX model, which incorporates the effects of metal speciation, affinity and competition of metals for binding sites. The combined toxicity of metals was quantified by the toxicity function (FTOX), a linear combination of the amount of metal binding to non-specific ligand sites (vi) and a toxicity coefficient (αi). Observed body concentrations of Ni and Co in the animals correlated well with the WHAM-calculated amounts binding to humic acid, supporting the use of humic acid as a surrogate for metal binding sites of E. crypticus. The toxicity of metals at different exposure times was well predicted by the WHAM-FTOX model. The derived αNi increased with time and reached equilibrium after approximately 14d, while αCo remained almost independent of time. This suggests for Ni more time is needed than for Co to reach equilibrium of body concentrations, so the toxicity of Ni is much more time-dependent. The WHAM-FTOX model provides a new tool for evaluating the potential mixture toxicity of metals to soil organisms in a dynamic environment. However, as αi varied with exposure time, caution is warranted when using the parameters estimated from acute toxicity experiments for predicting the chronic toxicity of metal mixtures., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Biotic ligand models (BLMs) predicting metal toxicity for a fixed exposure time are available, but it is uncertain how to extrapolate predictions to a dynamic environment with time-variable exposure. Three BLM-based models were developed to describe change of Ni toxicity to Enchytraeus crypticus in time. These models assumed that: (a) biotic ligand binding constants of Ni and competing cations (K(NiBL) and K(CBL)) and the fraction of biotic ligands occupied by Ni to produce 50% mortality (f50) are fixed with time, (b) K(NiBL) and K(CBL) remain constant while f50 varies with time, and (c) K(NiBL), K(CBL), and f50 are all time-dependent. Model (a) successfully described the 7-d toxicity of Ni but failed in explaining Ni toxicity at longer exposure times. Both models (b) and (c) well described Ni toxicity, within a factor of 2, at varying solution chemistries and different exposure times. This shows that the acute BLM cannot directly be applied for predicting chronic metal toxicity and that some BLM parameters may vary with time. Our findings provide plausible explanations for differences in mechanisms of acute and chronic toxicity, offering a framework for incorporating toxicokinetic and toxicodynamic processes in describing Ni toxicity in time., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

This study aimed at assessing the photodegradation of the insecticide chlorantraniliprole (CAP) in deionized water and in tap water amended with humic acids and nitrate. Photolysis was carried out under simulated solar or UV-A light. CAP (39 μM) photodegradation was slightly faster in tap water than in deionized water with half lives of 4.1 and 5.1 days, respectively. Photodegradation rate of CAP was hardly affected by humic acids (up to 100 mg L(-1)) and nitrate. Photodegradation pattern was different in slightly acidic (pH=6.1) deionized water compared to basic (pH=8.0) tap water. Four main degradation products have been isolated and characterized spectroscopically, and crystal structure was recorded for the first two photodegradation products. CAP also degraded in the dark controls, but only at basic pH (23% loss at pH 8.0 in tap water after 6 days), resulting in the formation of one single degradation product. Our study shows that the degradation of chlorantraniliprole in water is a combination of chemical and photochemical reactions, which are highly dependent on the pH of the solution., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Quantitative structure-activity relationships (QSARs) are an established tool in environmental risk assessment and a valuable alternative to the exhaustive use of test animals under REACH. In this study a QSAR was developed for the toxicity of a series of six chloroanilines to the soil-dwelling collembolan Folsomia candida in standardized natural LUFA2.2 soil. Toxicity endpoints incorporated in the QSAR were the concentrations causing 10% (EC10) and 50% (EC50) reduction in reproduction of F. candida. Toxicity was based on concentrations in interstitial water estimated from nominal concentrations in the soil and published soil-water partition coefficients. Estimated effect concentrations were negatively correlated with the lipophilicity of the compounds. Interstitial water concentrations for both the EC10 and EC50 for four compounds were determined by using solid-phase microextraction (SPME). Measured and estimated concentrations were comparable only for tetra- and pentachloroaniline. With decreasing chlorination the disparity between modelled and actual concentrations increased. Optimisation of the QSAR therefore could not be accomplished, showing the necessity to move from total soil to (bio)available concentration measurements., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Glucosinolates are compounds produced by commercial crops which can hydrolyse in a range of natural toxins that may exert detrimental effects on beneficial soil organisms. This study examined the effects of 2-phenylethyl isothiocyanate and 3-phenylpropionitrile on the survival and growth of the woodlouse Porcellio scaber exposed for 28 d. 2-Phenylethyl isothiocyanate dissipated from the soil with half-lives ranging from 19 to 96 h. Exposure through soil showed toxic effects only on survival. The LC50s after 28 d were significantly different at 65.3 mg kg(-1) for 2-phenylethyl isothiocyanate and 155 mg kg(-1) for 3-phenylpropionitrile. A toxicokinetic-toxicodynamic (TKTD) approach, however, revealed that both compounds in fact have very similar effect patterns. The TKTD model was better suited to interpret the survival data than descriptive dose-response analysis (LC(x)), accounting for the fast dissipation of the compounds in the soil. Found effects were within environmentally relevant concentrations. Care should therefore be taken before allowing these natural toxins to enter soil ecosystems in large quantities., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Enchytraeids are ecologically relevant soil organisms, due to their activity in decomposition and bioturbation in many soil types worldwide. The enchytraeid reproduction test (ERT) guidelines ISO 16387 and OECD 220 are exclusive to the genus Enchytraeus and recommend using the species E. albidus with a 6-week test period. The suggested alternative, E. crypticus has a shorter generation time which may enable the ERT to be twice as fast. To confirm the suitability of a 3-week test period for E. crypticus, the toxicity of five chemicals, with distinct properties and modes of action, was assessed in LUFA 2.2 soil. In all controls the validity criteria were met, as survival of E. crypticus was above 92% and more than 772 juveniles were produced. The good performance supports its appropriateness as model species. Reproduction was more sensitive than survival, with only cadmium and 3,5-dichloroaniline causing significant lethal effects in the tested concentration ranges. The effect concentration causing 50% reduction in the number of juveniles (EC50) was 35 mg kg(-1) for cadmium, <1.0 mg kg(-1) for carbendazim, 145 mg kg(-1) for phenanthrene, 275 mg kg(-1) for pentachloroaniline and 102 mg kg(-1) for 3,5-dichloroaniline. To evaluate the sensitivity of E. crypticus, the present results were compared to literature data for E. albidus. In conclusion, E. crypticus is a suitable model species in soil ecotoxicology, with advantages such as good control performance and speed, leading to a reliable and faster ERT., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Metal pollution e.g. copper, in water bodies occurs worldwide. Although copper is an essential trace metal, at certain levels it is still considered as pollutant. The aim of this study was to investigate the effect of exposure concentration on copper bioaccumulation in marbled crayfish (Procambarus sp.) by determining uptake and elimination kinetics. Crayfish were exposed to sub-lethal copper concentrations (average measured concentrations of 0.031 and 0.38 mg Cu L(-1)) for 14 d and transferred to copper-free water for another 14 d. At different time points during the uptake and elimination phases copper concentrations were measured in five organs (exoskeleton, gills, muscle, ovaries and hepatopancreas). At 0.031 mg Cu L(-1), copper levels in the crayfish organs were not significantly increased compared to the control animals, suggesting effective regulation. Exposure to 0.38 mg Cu L(-1) did lead to not significantly increased copper levels in muscles and ovaries, while the gills and exoskeleton, which are in direct contact with the water, showed significantly higher copper concentrations. In these four organs, copper showed fast uptake kinetics with equilibrium reached within 10 d of exposure. Copper accumulation was highest in the hepatopancreas; uptake in this storage organ steadily increased with time and did not reach equilibrium within the 14-d exposure period. Copper accumulation levels in the marbled crayfish found in this study were hepatopancreas>gills>exoskeleton>muscle., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Mo bioaccumulation in the earthworm Eisenia andrei was determined after 28 d exposure in ten different European field soils (pH 4.4-7.8) and an artificial soil, freshly spiked with Na₂MoO₄ at concentrations between 3.2 and 3200 mg Mo kg⁻¹ dry soil. Three field soils were also tested after ageing for 11 months. Earthworm Mo concentrations generally levelled off at high exposure levels but in most soils showed a (nearly) linear increase with increasing soil concentrations in the lower, non-toxic range (below EC10 or NOEC for reproduction effects). Bioaccumulation (BAF) and Bioconcentration factors (BCF) were calculated as the ratio of earthworm concentration to soil and estimated porewater concentrations, respectively. BAFs (0.35-3.44) and BCFs (1.31-276) did not seem much affected by soil concentration, suggesting that earthworms are not capable of regulating their internal Mo concentrations. BAF was best predicted by ammonium oxalate-extractable iron (Fe(ox)) and phosphor (P(ox)) contents of the soils., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

This study aimed at determining the uptake and elimination kinetics of molybdenum in the earthworm Eisenia andrei, and the influence of soil properties on molybdenum bioaccumulation. Three natural and four artificial soils were spiked at concentrations of 10 and 100 microg Mo g(-1) dry soil. Earthworms were exposed individually to spiked soils and sampled at different time intervals for 21 d. Remaining earthworms were transferred to non-spiked soil to determine elimination, also for 21 d. Uptake and elimination rate constants and bioaccumulation factors (BAFs) were calculated using a one-compartment model, and related to total, water and CaCl2 extractable molybdenum concentrations in soil. Molybdenum was rapidly accumulated by the earthworms with uptake rate constants between 0.05 and 1.70 g dry soil g(-1) dry earthworm d(-1), and equilibrium generally was reached within 10 d. Molybdenum was not strongly bioaccumulated (BAF

Two reference models are commonly used to predict mixture toxicity, Concentration Addition and Independent Action. For accurately predicting mixture effects, both reference models need a full description of the dose-response curve for all single chemicals present in the mixture. We studied the mixture effect of nickel and chlorpyrifos on survival of the soil-dwelling collembolan Folsomia candida exposed for 7weeks in Lufa 2.2 soil. Especially chlorpyrifos toxicity showed extremely steep dose-response curves, making it impossible to fulfil the assumptions required by both reference models. Nevertheless, we showed that by monitoring the development of toxicity in time for both the single compounds and the mixtures it was possible to determine mixture effects. Even at exposure levels below the No Effect Concentration, chlorpyrifos was found to reduce nickel toxicity. In addition, this study clearly showed that toxicity is a dynamic process and that mixture effects may be dependent on exposure time., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Endocrine disrupting compounds (EDCs) have been studied due to their impact on human health and increasing awareness of their impact on wildlife species. Studies concerning the organ-specific molecular effects of EDC in invertebrates are important to understand the mechanisms of action of this class of toxicants but are scarce in the literature. We have used a dose/response approach to unravel the protein expression in different organs of isopods exposed to bisphenol A (BPA) and vinclozolin (Vz) and assess their potential use as surrogate species. Male isopods were exposed to a range of Vz or of BPA concentrations. After animal dissection, proteins were extracted from gut, hepatopancreas and testes. Protein profiles were analysed by electrophoresis and differentially expressed proteins were identified by MALDI mass spectrometry. EDCs affected proteins involved in the energy metabolism (arginine kinase), proteins of the heat shock protein family (Hsp70 and GRP78) and most likely microtubule dynamics (tubulin). Different proteins expressed at different concentrations in different organs are indicative of the organ-specific effects of BPA and Vz. Additionally, several proteins were up-regulated at lower but not higher BPA or Vz concentrations, bringing new data to the non-monotonic response curve controversy. Furthermore, our findings suggest that some common responses to EDCs in both vertebrates and invertebrates may exist.

Endocrine Disruptor Compounds (EDCs) have been largely studied concerning their effects on vertebrates. Nevertheless, invertebrates as targets for these chemicals have been neglected and few studies are available. Specifically for edaphic invertebrates, data concerning the effects of EDCs is residual. Influences of EDCs on the reproduction systems of these organisms, with consequences at the population level, are expected but have not been confirmed. This work aimed to study the effects of bisphenol A (BPA) and vinclozolin (Vz) on the reproduction of the terrestrial isopod Porcellio scaber. Isopods were coupled and exposed to increasing concentrations of Vz and BPA and the females' reproductive cycle followed for 56d. Both compounds elicited reproductive toxicity. Vz and BPA decreased female reproductive allocation. Vz reduced pregnancy duration; increased the abortion percentage; decreased the number of pregnancies; and decreased the number of juveniles per female while BPA increased abortions at the lowest and highest test concentrations. The reproductive endpoints presented in here are indicative of the possible impact that this type of compounds might have on isopod population dynamics, which may eventually lead to population decline., (2009 Elsevier Ltd. All rights reserved.)

Terrestrial avoidance behavior is proposed as a fast and cost-effective method for assessing effects of pesticides on earthworms. Tropical species however, have rarely been used in avoidance tests. Avoidance tests were performed with Perionyx excavatus, a tropical species, and Eisenia andrei as the standard species, using chlorpyrifos and carbofuran in artificial and natural soil. Earthworms were exposed to concentrations of 1-900 (chlorpyrifos) and 1-32 (carbofuran)mg a.i. kg(-1) dry soil in a two-chamber system under tropical conditions (26+/-2 degrees C, 48 h). No significant difference was found in the control tests comparing the two soils used, suggesting soil type did not affect the distribution of the worms. The results suggest a higher sensitivity of E. andrei, with EC50S for the effect on avoidance behavior for both pesticides being a factor of 2-3 lowers than for P. excavatus. Earthworm avoidance tests with local species should therefore be used with caution when applied as a tool for pesticide risk assessment in the tropics. Endpoints generated through avoidance tests in this study are shown to be less sensitive than reproduction and more sensitive than survival. This was further confirmed by literature data available. Earthworm avoidance tests therefore can only replace survival tests as an initial screening tool for risk assessment.

The ecotoxicological characterization of sewage sludge takes into account the additive, antagonistic and synergistic effects that occur as a result of multi-chemical interactions. Such an evaluation therefore is essential to complement the chemical analysis that, although required by law, is clearly insufficient. Using a tiered approach in the toxic evaluation of sewage sludge allows for characterization of toxicity in a timely manner. According to the literature, reproduction tests with Folsomia candida are suitable tools for the toxic assessment of organic sludges. Therefore, the inclusion of Collembola avoidance tests at a screening level (low tier), and acting as a trigger for longer-period tests (high tier; e.g. reproduction test), may provide a successful strategy, and may complement the currently proposed test battery. To evaluate the use of both avoidance and reproduction tests with collembolans in such a tiered approach, three sewage sludges (urban, olive and electroplating industries) were mixed in with a field-collected soil at different concentrations. Avoidance and reproduction tests were performed with the soil-sludge mixtures after 0, 4 and 12 weeks of incubation. The tests detected no toxicity in soil-sludge mixtures of urban and olive sludges at any incubation period. Mixtures with sludge from the electroplating industry induced toxicity only in the avoidance tests with freshly prepared and 4-week incubated samples. These results demonstrate the ability of Collembola avoidance tests to assess sewage sludge toxicity over time and its potential for hazardous sludge characterization at low tier levels.

Expansion of agriculture in the tropics has increased the use of pesticides that may affect the soil ecosystems. Few studies so far determined the effects of pesticides in the tropics and tropical risk assessment therefore often relies on data from temperate conditions. Hence we compared the toxicity of chlorpyrifos, carbofuran and carbendazim to the earthworm Eisenia andrei at two different temperatures reflecting temperate and tropical conditions. The toxicity of the three pesticides in both conditions decreased in the order carbendazim>carbofuran>chlorpyrifos. For chlorpyrifos and carbofuran, but not for carbendazim, survival was more sensitive at the higher temperature, probably due to increased earthworm activity. Sub-lethal effects (reproduction and growth) however, varied inconsistently with temperature and soil types. We conclude that toxicity of pesticides in tropics may not be predicted from data generated under temperate conditions, even within the same species.

Imidacloprid is a neonicotinoid insecticide with neurotoxic action that, as a possible alternative for commonly used organophosphorus pesticides, has gained registration in about 120 countries for use in over 140 agricultural crops. Only few data are available on its toxicity for soil invertebrates. We therefore assessed the effects of imidacloprid on survival, weight gain, feeding rate, total protein content, glutathione S-transferase activity (GST), and digestive gland epithelial thickness in juveniles and adults of the terrestrial isopod Porcellio scaber. After two weeks of feeding on imidacloprid-dosed food, weight gain (NOEC 5 microg/g dry food) and feeding rate (NOEC 10 microg/g) in juveniles, and feeding rate (NOEC<10 microg/g) and digestive gland epithelial thickness (NOEC<10 microg/g) in adults were most affected. In juveniles induction of GST activity and increase of total protein content per wet animal weight was detected at 5 microg/g dry food, whereas in adults a reduction of GST was observed at 25 microg/g (NOEC 10 microg/g). An estimate of actual intake rates suggests that imidacloprid affects isopods at similar exposure concentrations as insects. The toxicity of imidacloprid was similar to that of the organophosphorus pesticide diazinon, tested earlier using the same methods [Stanek, K., Drobne, D., Trebse, P., 2006. Linkage of biomarkers along levels of biological complexity in juvenile and adult diazinon fed terrestrial isopod (Porcellio scaber, Isopoda, Crustacea). Chemosphere 64, 1745-1752]. At actual environmental concentrations, diazinon poses a higher risk to P. scaber. Due to its increasing use in crop protection and higher persistence in soil, imidacloprid might however, be potentially more dangerous after long-term application. We conclude that toxicity testing with P. scaber provides relevant, repeatable, reproducible and comparable toxicity data that is useful for the risk assessment of pesticides in the terrestrial environment.

The effect of zinc on soil nitrification and composition of the microbial community in soil was investigated using a full factorial experiment with five zinc concentrations and four levels of biological complexity (microbes only, microbes and earthworms (Eisenia fetida), microbes and Italian ryegrass (Lolium multiflorum var. Macho), and microbes, ryegrass and earthworms). After 6 weeks of exposure, the activity of soil nitrifying bacteria was measured and the microbial community structure was characterized by phospholipid fatty acid (PLFA) analysis. Soil nitrification and several PLFA markers were significantly influenced by either zinc addition and/or the presence of earthworms or ryegrass, and one of the most pronounced changes was the increase of fungi and decrease of bacteria with increasing concentrations of zinc. Of particular interest, however, was the potential interaction between the presence of plants and/or earthworms and the effect of zinc, which the factorial study design allowed us to explore. Such an effect was observed in two cases: Earthworms reduced the positive effect of zinc on the fungal biomass (ANOVA, p=0.03), and the effect of earthworms on the soil nitrification activity depended on zinc concentration (ANOVA, p<0.05). The effect of earthworm presence was not very large, but it does show that multispecies tests might give information about metal toxicity or bioavailability that cannot be predicted from single-species tests.

Uptake and accumulation of three chlorobenzenes was studied in both biota (Enchytraeus crypticus) and 30 mum polydimethylsiloxane (PDMS) solid phase microextraction (SPME) fibers after exposure to spiked OECD soil. The OECD soil was spiked with three different concentrations of all contaminants. Uptake of all three chlorobenzenes in E. crypticus was fast and steady state levels were reached within 2-4 days. Also in the PDMS-SPME fibers uptake was very fast for all three compounds, with steady state levels reached after 1 day. Comparison of steady state levels in biota and in the PDMS-SPME fibers showed a relationship which was consistent over the range of concentrations of chlorobenzenes in soil and the difference in logKow. This shows that measuring the concentrations of hydrophobic chemicals in a hydrophobic phase such as PDMS can be used as a simple tool to estimate internal concentrations of these chemicals in biota exposed to complex matrices such as soil.