Toxicity of binary mixtures of Cu, Cr and As to the earthworm Eisenia andrei

Chromated copper arsenate (CCA) mixtures were used in the past for wood preservation, leading to large scale soil contamination. This study aimed at contributing to the risk assessment of CCA-contaminated soils by assessing the toxicity of binary mixtures of copper, chromium and arsenic to the earthwormEisenia andreiin OECD artificial soil. Mixture effects were related to reference models of Concentration Addition (CA) and Independent Action (IA) using the MIXTOX model, with effects being related to total and available (H2O and 0.01 M CaCl(2)extractable) concentrations in the soil. Since only in mixtures with arsenic dose-related mortality occurred (LC(50)92.5 mg/kg dry soil), it was not possible to analyze the mixture effects on earthworm survival with the MIXTOX model. EC(50)s for effects of Cu, Cr and As on earthworm reproduction, based on total soil concentrations, were 154, 449 and 9.1 mg/kg dry soil, respectively. Effects of mixtures were mainly antagonistic when related to the CA model but additive related to the IA model. This was the case when mixture effects were based on total and H2O-extractable concentrations; when based on CaCl2-extractable concentrations effects mainly were additive related to the CA model except for the Cr-As mixture which acted antagonistically. These results suggest that the CCA components do interact leading to a reduced toxicity when present in a mixture.Peer reviewe

An uptake and elimination kinetics approach to assess the bioavailability of chromium, copper, and arsenic to earthworms (Eisenia andrei) in contaminated field soils

The aim of this study was to determine the bioavailability of metals in field soils contaminated with chromated copper arsenate (CCA) mixtures. The uptake and elimination kinetics of chromium, copper, and arsenic were assessed in the earthworm Eisenia andrei exposed to soils from a gradient of CCA wood preservative contamination near Hartola, Finland. In soils contaminated with 1480–1590 mg Cr/kg dry soil, 642–791 mg Cu/kg dry soil, and 850–2810 mg Ag/kg dry soil, uptake and elimination kinetics patterns were similar for Cr and Cu. Both metals were rapidly taken up and rapidly excreted by Eisenia andrei with equilibrium reached within 1 day. The metalloid As, however, showed very slow uptake and elimination in the earthworms and body concentrations did not reach equilibrium within 21 days. Bioaccumulation factors (BAF) were low for Cu and Cr (Peer reviewe

No effect of selected engineered nanomaterials on reproduction and survival of the springtail Folsomia candida

Although the number of studies on engineered nanomaterial (NM) toxicity to soil invertebrates is increasing, only a few studies have reported toxicity of NMs to soil dwelling model species, such as the invertebrate Folsomia candida. The main objective of this study was to determine the toxicity of five different engineered NMs (WCCo, CuO, Fe2O3, organic pigment and MWCNTs) for the springtail F. candida. Copper(ii), cobalt and iron chlorides were taken as positive controls. A standardized OECD test was used to measure effects on reproduction and survival, and toxicity was related to metal concentrations in soil and pore water. None of the NMs exerted adverse effects on springtail reproduction and survival at concentrations up to 6400 mg per kg dry soil, whereas the Cu, Co and Fe chlorides resulted in 50% decline in springtail reproduction at 981, 469 and 569 mg metal ion per kg dry soil, respectively. The absence of toxicity of the NMs could partly be explained by the low porewater metal concentrations, suggesting low solubility or slow solubilisation. The fate of engineered NMs in soil is rather complex but needs better understanding to facilitate predicting exposure of soil organisms

Resilience of soil microbial communities to metals and additional stressors : DNA-based approaches for assessing "Stress-on-Stress" responses

International audienceMany microbial ecology studies have demonstrated profound changes in community composition caused by environmental pollution, as well as adaptation processes allowing survival of microbes in polluted ecosystems. Soil microbial communities in polluted areas with a long-term history of contamination have been shown to maintain their function by developing metal-tolerance mechanisms. In the present work, we review recent experiments, with specific emphasis on studies that have been conducted in polluted areas with a long-term history of contamination that also applied DNA-based approaches. We evaluate how the "costs" of adaptation to metals affect the responses of metal-tolerant communities to other stress factors ("stress-on-stress"). We discuss recent studies on the stability of microbial communities, in terms of resistance and resilience to additional stressors, focusing on metal pollution as the initial stress, and discuss possible factors influencing the functional and structural stability of microbial communities towards secondary stressors. There is increasing evidence that the history of environmental conditions and disturbance regimes play central roles in responses of microbial communities towards secondary stressors

Toxicokinetics of Zn and Cd in the earthworm Eisenia andrei exposed to metal-contaminated soils under different combinations of air temperature and soil moisture content

This study evaluated how different combinations of air temperature (20 °C and 25 °C) and soil moisture content (50% and 30% of the soil water holding capacity, WHC), reflecting realistic climate change scenarios, affect the bioaccumulation kinetics of Zn and Cd in the earthworm Eisenia andrei. Earthworms were exposed for 21 d to two metal-contaminated soils (uptake phase), followed by 21 d incubation in non-contaminated soil (elimination phase). Body Zn and Cd concentrations were checked in time and metal uptake (k1) and elimination (k2) rate constants determined; metal bioaccumulation factor (BAF) was calculated as k1/k2. Earthworms showed extremely fast uptake and elimination of Zn, regardless of the exposure level. Climate conditions had no major impacts on the bioaccumulation kinetics of Zn, although a tendency towards lower k1 and k2 values was observed at 25 °C + 30% WHC. Earthworm Cd concentrations gradually increased with time upon exposure to metal-contaminated soils, especially at 50% WHC, and remained constant or slowly decreased following transfer to non-contaminated soil. Different combinations of air temperature and soil moisture content changed the bioaccumulation kinetics of Cd, leading to higher k1 and k2 values for earthworms incubated at 25 °C + 50% WHC and slower Cd kinetics at 25 °C + 30% WHC. This resulted in greater BAFs for Cd at warmer and drier environments which could imply higher toxicity risks but also of transfer of Cd within the food chain under the current global warming perspective.publishe

Toxicokinetics of silver nanoparticles in the mealworm Tenebrio molitor exposed via soil or food

Silver nanoparticles (AgNPs) may reach the soil compartment via sewage sludge or nanoagrochemical applications. Understanding how NPs interact with biological systems is crucial for an accurate hazard assessment. Therefore, this study aimed at determining the Ag toxicokinetics in the mealworm Tenebrio molitor, exposed via Lufa 2.2 soil or via food to different Ag forms (uncoated 50 nm AgNPs, paraffin coated 3–8 nm and PVP-stabilised 60 nm, Ag2S NPs 20 nm, and ionic Ag). Mealworms were exposed for 21 days followed by a 21-day elimination phase (clean soil/food). A one-compartment kinetics model with inert fraction (simulating a storage compartment, where detoxified forms are located) was used to describe Ag accumulation. Fully understanding the uptake route in mealworms is difficult. For that reason several approaches were used, showing that food, soil and pore water all are valid uptake routes, but with different importance. Silver taken up from soil pore water or from soil showed to be related to Ag dissolution in soil pore water. In general, the uptake and elimination rate constants were similar for 3–8 nm and 60 nm AgNPs and for AgNO3, but significantly different for the uncoated 50 nm AgNPs. Upon food exposure, uptake rate constants were similar for 50 nm AgNPs and AgNO3, while those for 60 nm and 3–8 nm AgNPs and for Ag2S NPs also grouped together. NP exposure in soil appeared more difficult to characterize, with different patterns obtained for the different NPs. But it was evident that upon soil or food exposure, particle characteristics highly affected Ag bioavailability and bioaccumulation. Although Ag2S NPs were taken up, their elimination was faster than for other Ag forms, showing the lowest inert fraction. The significantly different elimination rate constants suggest that the mechanism of elimination may not be the same for different AgNPs either

The use of gene expression to unravel the single and mixture toxicity of abamectin and difenoconazole on survival and reproduction of the springtail Folsomia candida

Pesticides risk assessments have traditionally focused on the effects on standard parameters, such as mortality, reproduction and development. However, one of the first signs of adverse effects that occur in organisms exposed to stress conditions is an alteration in their genomic expression, which is specific to the type of stress, sensitive to very low contaminant concentrations and responsive in a few hours. The aim of the present study was to evaluate the single and binary mixture toxicity of commercial products of abamectin (Kraft® 36 EC) and difenoconazole (Score® 250 EC) to Folsomia candida. Laboratory toxicity tests were conducted to access the effects of these pesticides on springtail survival, reproduction and gene expression. The reproduction assays gave EC50 and EC10 values, respectively, of 6.3 and 1.4 mg a.s./kg dry soil for abamectin; 1.0 and 0.12 mg a.s./kg dry soil for Kraft® 36 EC; and 54 and 23 mg a.s./kg dry soil for Score® 250 EC. Technical difenoconazole did not have any effect at the concentrations tested. No significant differences in gene expression were found between the abamectin concentrations tested (EC10 and EC50) and the solvent control. Exposure to Kraft® 36 EC, however, significantly induced Cyp6 expression at the EC50 level, while VgR was significantly downregulated at both the EC10 and EC50. Exposure to the simple pesticide mixture of Kraft® 36 EC + Score® 250 EC caused significant up regulation of ABC transporter, and significant down regulation of VgR relative to the controls. GABA receptor also showed significant down-regulation between the EC10 and EC50 mixture treatments. Results of the present study demonstrate that pesticide-induced gene expression effects precede and occur at lower concentrations than organism-level responses. Integrating "omic" endpoints in traditional bioassays may thus be a promising way forward in pesticide toxicity evaluations

Phenotypic and transcriptional responses associated with multi-generation exposure of Folsomia candida to engineered nanomaterials

Sublethal effects of toxicants may cumulate over time and become apparent only when test organisms are exposed for multiple generations. In this study we determined phenotypic effects and transcriptional responses in the parthenogenetic soil invertebrate Folsomia candida over four generations, followed by two generations of recovery. Animals were exposed to two metal-based nanomaterials (NMs): copper oxide (CuO) and tungsten carbide–cobalt (WCCo), both homogenously mixed in with the soil. Survival and reproduction were not affected in any of four consecutive generations of F. candida exposed to CuO-NM at concentrations as high as 6400 mg Cu per kg dry LUFA 2.2 soil. WCCo-NM affected reproduction and survival from the third generation onwards, with EC50 values between 2400 and 5600 mg NM per kg dry soil, but recovery was seen in recovery generations 1 and 2 when kept in clean soil. Histological investigations showed that WCCo-NM (3200 mg kg−1) induced tissue damage and loss of villi from the gut epithelial cells. Expression of four target genes known to be responsive to stress were investigated by quantitative PCR at different exposure levels and in different generations. Expression of all genes was significantly affected by NMs even though exposures were below toxic threshold concentrations. In addition, gene expression did not always return to control levels during consecutive recovery over two generations in clean soil. This shows that gene expression assays can detect physiological alterations cumulating from one generation to the next initially without visible effects on phenotypic variables such as reproduction. The possibility of multi-generation carry-over of sublethal toxicity needs more attention in environmental risk assessment

Transcriptome assembly and microarray construction for Enchytraeus crypticus, a model oligochaete to assess stress response mechanisms derived from soil conditions

The soil worm Enchytraeus crypticus (Oligochaeta) is an ecotoxicology model species that, until now, was without genome or transcriptome sequence information. The present research aims at studying the transcriptome of Enchytraeus crypticus, sampled from multiple test conditions, and the construction of a high-density microarray for functional genomic studies. Over 1.5 million cDNA sequence reads were obtained representing 645 million nucleotides. After assembly, 27,296 contigs and 87,686 singletons were obtained, from which 44% and 25% are annotated as protein-coding genes, respectively, sharing homology with other animal proteomes. Concerning assembly quality, 84% of the contig sequences contain an open reading frame with a start codon while E. crypticus homologs were identified for 92% of the core eukaryotic genes. Moreover, 65% and 77% of the singletons and contigs without known homologs, respectively, were shown to be transcribed in an independent microarray experiment. An Agilent 180 K microarray platform was designed and validated by hybridizing cDNA from 4 day zinc- exposed E. crypticus to the concentration corresponding to 50% reduction in reproduction after three weeks (EC50). Overall, 70% of all probes signaled expression above background levels (mean signal + 1x standard deviation). More specifically, the probes derived from contigs showed a wider range of average intensities when compared to probes derived from singletons. In total, 522 significantly differentially regulated transcripts were identified upon zinc exposure. Several significantly regulated genes exerted predicted functions (e.g. zinc efflux, zinc transport) associated with zinc stress. Unexpectedly, the microarray data suggest that zinc exposure alters retro transposon activity in the E. crypticus genome. An initial investigation of the E. crypticus transcriptome including an associated microarray platform for future studies proves to be a valuable resource to investigate functional genomics mechanisms of toxicity in soil environments and to annotate a potentially large number of lineage specific genes that are responsive to environmental stress conditions