Copper toxicity to earthworms: A comprehensive review and meta-analysis.

Copper can accumulate in agricultural topsoil through the use of Cu-based fungicides, which may harm soil organisms such as earthworms. This study aimed at reviewing the effects of copper on earthworms at different levels of biological organization, and to determine critical values of copper toxicity to earthworms using a meta-analysis and accounting for lethal and sub-lethal effects and different earthworm species and exposure conditions. Endpoints at the sub-individual level were more sensitive than at higher levels of organization. At the individual level, the most sensitive endpoints were reproduction and growth (hatching success, hatchling growth). Hormetic growth was clearly recognized at copper concentrations less than 80 mg kg-1 in dry soil. However, effects at the sub-individual level already occurred at lower concentrations. Considering all the exposure conditions, the calculated weighted means were 113 mg Cu kg-1 dry soil (95% CI -356; 582) for the LC50 (lethal concentration for 50% of the exposed individuals), 94.6 mg Cu kg-1 dry soil (95% CI 14.0; 175) for the EC50 reproduction, and 144 mg Cu kg-1 dry soil (95% CI -12.6; 301) for the EC50 growth or weight change. When accounting for the origin of the soil, earthworms were five times more sensitive to copper (LC50) in natural than in artificial soils. The different factors affecting Cu toxicity to earthworms explain the high variability of these values, making it difficult to derive thresholds. However, considering the potential negative effects of copper on earthworms, attention should be given to the more sustainable use of human-contributed copper in agricultural soils.

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.

Impact of chromium on the soil invertebrate model Enchytraeus crypticus (Oligochaeta) in standard reproduction and full life cycle tests.

Chromium (Cr) has many applications in industry and day-to-day life, entering the terrestrial environment mostly from anthropogenic sources. Despite the fact that Cr is essential, it can be harmful in biota at high concentrations. Full life cycle tests (FLCt) are scarce, representing a gap for soil ecotoxicology. Hence, the effects of Cr were studied using the soil organism Enchytraeus crypticus in the 46-day FLCt and the standard 21-day enchytraeid reproduction test (ERT). FLCt hatching (day-11) and time to reach maturity (day 22-25) were the most sensitive endpoints, representing a delay, partly recovered with time. Reproduction was reduced to similar levels in both the ERT and the FLCt (EC50∼320 mg Cr/kg), but survival was more affected in the ERT (LC50 = 377 mg Cr/kg) than in the FLC (LC50 = 467 mg Cr/kg). This could be due to the induction of stress response mechanisms at earlier life stages (cocoons/juveniles), providing protection to toxicity in a later stage (adults). FLCt results provided considerably more details and data points, and the additional endpoints allowed to explain the source of observed effects, e.g. a direct impact on fecundity and not only due to adult mortality. The estimated population growth curves confirmed the significant effect of 320 mg Cr/kg and onwards, reflecting the impact of Cr on both cocoon production and juvenile performance, with follow-up consequences for reproduction. An Adverse Outcome Pathway was drafted. The FLCt design is recommended as an upgrade of the current standard ERT, which could be integrated into the existing OECD ERT test guideline.

Toxicity to Eisenia andrei and Folsomia candida of a metal mixture applied to soil directly or via an organic matrix.

Regulatory limits for chemicals and ecological risk assessment are usually based on the effects of single compounds, not taking into account mixture effects. The ecotoxicity of metal-contaminated sludge may, however, not only be due to its metal content. Both the sludge matrix and the presence of other toxicants may mitigate or promote metal toxicity. To test this assumption, the toxicity of soils recently amended with an industrial sludge predominantly contaminated with chromium, copper, nickel, and zinc and soils freshly spiked with the same mixture of metals was evaluated through earthworm (Eisenia andrei) and collembolan (Folsomia candida) reproduction tests. The sludge was less toxic than the spiked metal mixture for E. andrei but more toxic for F. candida. Results obtained for the earthworms suggest a decrease in metal bioavailability promoted by the high organic matter content of the sludge. The higher toxicity of the sludge for F. candida was probably due to the additive toxic effect of other pollutants.

A Kinetic Approach for Assessing the Uptake of Ag from Pristine and Sulfidized Ag Nanomaterials to Plants.

Nanomaterials (NMs) are thermodynamically unstable by nature, and exposure of soil organisms to NMs in the terrestrial environment cannot be assumed constant. Thus, steady-state conditions may not apply to NMs, and bioaccumulation modeling for uptake should follow a dynamic approach. The one-compartment model allows the uptake and elimination of a chemical to be determined, while also permitting changes in exposure and growth to be taken into account. The aim of the present study was to investigate the accumulation of Ag from different Ag NM types (20 nm Ag0 NMs, 50 nm Ag0 NMs, and 25 nm Ag2 S NMs) in the crop plant wheat (Triticum aestivum). Seeds were emerged in contaminated soils (3 or 10 mg Ag/kg dry soil, nominal) and plants grown for up to 42 d postemergence. Plant roots and shoots were collected after 1, 7, 14, 21, and 42 d postemergence; and total Ag was measured. Soil porewater Ag concentrations were also measured at each sampling time. Using the plant growth rates in the different treatments and the changing porewater concentrations as parameters, the one-compartment model was used to estimate the uptake and elimination of Ag from the plant tissues. The best fit of the model to the data included growth rate and porewater concentration decline, while showing elimination of Ag to be close to zero. Uptake was highest for Ag0 NMs, and size did not influence their uptake rates. Accumulation of Ag from Ag2 S NMs was lower, as reflected by the lower porewater concentrations. Environ Toxicol Chem 2021;40:1861-1872. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Developmental toxicity of endocrine disrupters bisphenol A and vinclozolin in a terrestrial isopod.

Studies of the effects of endocrine-disrupting compounds (EDCs) on invertebrates are still largely underrepresented. This work aims to fill this gap by assessing the effects of bisphenol A (BPA) and vinclozolin (Vz) on the terrestrial isopod Porcellio scaber (common rough woodlouse). Male adult and sexually undifferentiated juvenile woodlice were exposed to the toxicants. Effects on molting regime and growth were investigated independently for males and female woodlice after sexual differentiation. Both chemicals elicited developmental toxicity to P. scaber by causing overall decreased growth. Nevertheless, BPA induced molting, whereas Vz delayed it. Although the LC50 values for juvenile and adult survival were fairly similar, juvenile woodlice showed an increased chronic sensitivity to both chemicals, and female woodlice were most the sensitive to BPA. We recommend the use of adults, juveniles, female, and male woodlice, as well as a large range of toxicant concentrations, to provide valuable information regarding differential dose responses, effects, and threshold values for EDCs.

Scaling relationships based on partition coefficients and body sizes have similarities and interactions.

The LC(50) of compounds with a similar biological effect, at a given exposure period, is frequently plotted log-log against the octanol-water partition coefficient and a straight line is fitted for interpolation purposes. This is also frequently done for physiological properties, such as the weight-specific respiration rate, as function of the body weight of individuals. This paper focuses on the remarkable observation that theoretical explanations for these relationships also have strong similarities. Both can be understood as result of the covariation of the values of parameters of models of a particular type for the underlying processes, while this covariation follows logically from the model structure. The one-compartment model for the uptake and elimination of compounds by organisms is basic to the BioConcentration Factor (BCF), or the partition coefficient; the standard Dynamic Energy Budget model is basic to the (ultimate) body size. The BCF is the ratio of the uptake and the elimination rates; the maximum body length is the ratio of the assimilation (i.e. uptake of resources) and the maintenance (i.e. use of resources) rates. This paper discusses some shortcomings of descriptive approaches and conceptual aspects of theoretical explanations. The strength of the theory is in the combination of why metabolic transformation depends both on the BCF and the body size. We illustrate the application of the theory with several data sets from the literature.

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

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

Bioavailability of contaminants estimated from uptake rates into soil invertebrates.

It is often argued that the concentration of a pollutant inside an organism is a good indicator of its bioavailability, however, we show that the rate of uptake, not the concentration itself, is the superior predictor. In a study on zinc accumulation and toxicity to isopods (Porcellio scaber) the dietary EC(50) for the effect on body growth was rather constant and reproducible, while the internal EC(50) varied depending on the accumulation history of the animals. From the data a critical value for zinc accumulation in P. scaber was estimated as 53 microg/g/wk. We review toxicokinetic models applicable to time-series measurements of concentrations in invertebrates. The initial slope of the uptake curve is proposed as an indicator of bioavailability. To apply the dynamic concept of bioavailability in risk assessment, a set of representative organisms should be chosen and standardized protocols developed for exposure assays by which suspect soils can be evaluated.

Influence of soil properties on the bioaccumulation and effects of arsenic in the earthworm Eisenia andrei.

This study aimed at assessing the influence of soil properties on the uptake and toxicity effects of arsenic in the earthworm Eisenia andrei exposed for 4 weeks to seven natural soils spiked with different arsenic concentrations. Water-soluble soil concentrations (AsW) and internal As concentrations in the earthworms (AsE) were greatly different between soils. These two variables were highly correlated and were key factors in earthworm toxicity response. AsW was explained by some soil properties, such as the pH, calcium carbonate content, ionic strength, texture or oxide forms. Toxicity showed a clear variation between soils, in some cases without achieving 50 % adverse effect at the highest As concentration added (600 mg kg(-1)). Nevertheless, soil properties did not show, in general, a high relation with studied toxicity endpoints, although the high correlation with AsW could greatly reduce indirectly As bioavailability and toxicity risk for earthworms. Obtained results suggest that soil properties should be part of the criteria to establishing thresholds for contaminated soils because they will be key in controlling As availability and thus result in different degrees of toxicity.

Effect of soil properties on the toxicity of Pb: assessment of the appropriateness of guideline values.

Soil contamination with lead is a worldwide problem. Pb can cause adverse effects, but its mobility and availability in the terrestrial environment are strongly controlled by soil properties. The present study investigated the influence of different soil properties on the solubility of lead in laboratory spiked soils, and its toxicity in three bioassays, including Lactuca sativa root elongation and Vibrio fischeri illumination tests applied to aqueous extracts and basal soil respiration assays. Final aim was to compare soil-dependent toxicity with guideline values. The L. sativa bioassay proved to be more sensitive to Pb toxicity than the V. fischeri and soil respiration tests. Toxicity was significantly correlated with soil properties, with soil pH, carbonate and organic carbon content being the most important factors. Therefore, these variables should be considered when defining guideline values.

Risk assessment of heavy metal pollution for detritivores in floodplain soils in the Biesbosch, The Netherlands, taking bioavailability into account.

Floodplains of the European rivers Rhine and Meuse are heavily polluted. We investigated the risk of heavy metal pollution (Cd, Cu, Pb, Zn) for detritivores living in a floodplain area, the Biesbosch, the Netherlands, affected by these rivers. Total soil, pore water and 0.01 M CaCl(2) extractable concentrations and concentrations in plant leaves, earthworms, isopods and millipedes were measured in two sites and compared with literature data to assess possible risks. Based on total metal concentrations in soil, serious effects on detritivores were expected. However, 0.01 M CaCl(2) extractable, pore water and plant leaf concentrations were similar to metal concentrations found in unpolluted areas. Concentrations of Cu and Cd in earthworms and Cu in millipedes were higher in the Biesbosch than in animals from reference areas. All other measured concentrations of heavy metals in earthworms, isopods and millipedes were similar to the ones found in reference areas. Despite high total soil concentrations, effects of Zn, Cu, Pb and Cd pollution on isopods are therefore not expected, while millipedes may only be affected by Cu. Since Cu and Cd levels in earthworms were increased compared to animals in unpolluted soils, this faunal group seems to be most at risk. Given the engineering role of earthworms in ecosystems, effects on the ecological functioning of floodplain soils therefore cannot be excluded.

Solid phase microextraction as a tool to predict internal concentrations of soil contaminants in terrestrial organisms after exposure to a laboratory standard soil.

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.

Copper kinetics and internal distribution in the marbled crayfish (Procambarus sp.).

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.

Narcotic mechanisms of acute toxicity of chlorinated anilines in Folsomia candida (Collembola) revealed by gene expression analysis.

In order to clarify the mechanisms of reproductive toxicity in a QSAR approach, the transcriptional signatures upon the 2 day exposure to the 28 days EC50 of a series of increasingly chlorinated aniline compounds and 1,2,3,4-tetrachlorobenzene were measured in Folsomia candida. In general, the transcriptional patterns associated with all compounds revealed toxicity at the cellular membranes and hence components of narcosis type I, but a principal component analysis revealed a deviating response by the pentachloroaniline and 2,3,5,6-tetrachloroaniline exposure. Moreover the expression of a subset of mainly biotransformation related genes showed a significant relationship with the logK(ow,) which stresses the presence of narcosis type I. This was confirmed by GO term enrichment at the level of cellular component. Genes involved in the three phases of xenobiotic biotransformation exhibited strict compound specific transcription patterns, which may reflect biotransformation processes in F. candida. Additional toxic mechanisms were especially observed for the 2,3,5,6-tetrachloroaniline, which possible works as an uncoupler or inhibitor of electron transport systems, which is revealed by the up-regulation of genes that encode different members of the electron transport chain. The aniline and 2,3,4-trichloroaniline exposure caused the induction of genes in the ROS defense system. Additional toxicity mechanisms were less clear, but they include the attack of microbial pathogens for the six other compounds and for 2,3,5,6-tetrachloroaniline an effect on mitochondrial protein folding.

Reproductive toxicity of the endocrine disrupters vinclozolin and bisphenol A in the terrestrial isopod Porcellio scaber (Latreille, 1804).

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.

Elimination kinetic model for organic chemicals in earthworms.

Mechanistic understanding of bioaccumulation in different organisms and environments should take into account the influence of organism and chemical depending factors on the uptake and elimination kinetics of chemicals. Lipophilicity, metabolism, sorption (bioavailability) and biodegradation of chemicals are among the important factors that may significantly affect the bioaccumulation process in soil organisms. This study attempts to model elimination kinetics of organic chemicals in earthworms by accounting for the effects of both chemical and biological properties, including metabolism. The modeling approach that has been developed is based on the concept for simulating metabolism used in the BCF base-line model developed for predicting bioaccumulation in fish. Metabolism was explicitly accounted for by making use of the TIMES engine for simulation of metabolism and a set of principal transformations. Kinetic characteristics of transformations were estimated on the basis of observed kinetics data for the elimination of organic chemicals from earthworms.

The use of Collembola avoidance tests to characterize sewage sludges as soil amendments.

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.

Ambient trace element background concentrations in soils and their use in risk assessment.

The definition of ambient background concentrations (ABCs) is used in this study to assess the potential environmental risk of trace elements in soils and parent materials from Granada, Spain. Two different layers of soil (0-20 and 20-40 cm) and parent material samples were collected at 93 sites. From cumulative frequency distribution curves, ABCs for As, Co, Cu, Cr, Ni, Pb and Zn were estimated at 3.5-20, 7-23, 13-25.6, 29-66, 7-20, 15-36, and 5.5-76 mg kg(-1), respectively. Tukey box-plots were used to discriminate different concentration classes and identify potentially contaminated sites. Weakly-weathered soils (Entisols) over carbonate materials showed the lowest background contents, the most developed soils (Alfisols) over metamorphic rocks the highest ones. Outliers were mainly found near a former iron mine where arsenic concentrations were by far exceeding the corresponding regional ABC. These soils were however, not toxic to Escherichia coli and Vibrio fischeri. The prediction of site-specific ABCs together with bioavailability and toxicity assessment is a valuable tool for giving further insight into the risk of trace elements in soils.

Comparative chronic toxicity of homo- and heterocyclic aromatic compounds to benthic and terrestrial invertebrates: Generalizations and exceptions.

The aim of the present study was to elucidate consistent patterns in chronic polycyclic aromatic compound (PAC) toxicity to soil and sediment inhabiting invertebrates. Therefore we examined our experimental dataset, consisting of twenty-one chronic effect concentrations for two soil invertebrates (Folsomia candida and Enchytraeus cripticus) and two sediment invertebrates (Lumbriculus variegatus and Chironomus riparius) exposed to six PACs (two homocyclic isomers, anthracene and phenanthrene; two azaarene isomers: acridine and phenanthridine; and two azaarene transformation products, acridone and phenanthridone). In order to determine if effect concentrations were accurately predicted by existing toxicity-K(ow) relationships describing narcosis, chronic pore water effect concentrations were plotted jointly against logK(ow). Fifteen of the twenty-one effect concentrations (71%) were above the lower limit for narcosis, showing that narcosis was the main mode of action for the majority of the tested homo- and heterocyclic PACs during chronic exposure. Toxicity of all tested compounds to soil organisms was accurately described by the toxicity-K(ow) relationship. However, for the sediment invertebrates exposed to some of the tested heterocyclic PACs deviations from narcosis were identified, related to specific physicochemical properties of the test compounds and/or species specific sensitivities. It is concluded that existing toxicity-K(ow) relationships describing narcosis in some cases underestimate chronic PAC toxicity to sediment inhabiting invertebrates.