International audience | In recent decades, stormwater management has developed to allow stormwater to infiltrate directly into the soils instead of being collected and routed to sewer systems. However, during infiltration, stormwater creates a sediment deposit at the soil surface as the result of high loads of suspended particles (including pollutants), leading to the settlement of sedimentary layers prone to colonization by plants and earthworms. This study aims to investigate the earthworm communities of a peculiar infiltration basin and investigate the influence of edaphic conditions (water content, organic matter content, pH, height of sediment) and of persistent organic pollutants (POPs: PCBs, PCDDs and PCDFs) on these earthworms. Attention was paid to their age (juveniles or adults) and their functional group (epigeic, endogeic, anecic). We found that the earthworm abundance was mostly driven by edaphic conditions, with only a slight impact of POPs, with a significant negative impact of PCBDLno for juveniles and endogeic, and PCDDs for epigeic. On the contrary, the height of the sediment and the water content are beneficial for their presence and reproduction. Furthermore, POPs contents are also linked to physicochemical parameters of the sediment. Bioaccumulation was clearly revealed in the studied site but does not differ between juveniles and adults, except for PCDDs. Conversely, BAF values seemed to vary between functional groups, except for PCBDL non-ortho. It strongly varies with the family types (PCBs versus PCCD/Fs) and between congeners within the same family, with specific strong bioaccumulation for a few congeners.

International audience | Assessment of wildlife exposure to organophosphorus (OP) pesticides generally involves the measurement of cholinesterase (ChE) inhibition, and complementary biomarkers (or related endpoints) are rarely included. Herein, we investigated the time course inhibition and recovery of ChE and carboxylesterase (CE) activities in the earthworm Lumbricus terrestris exposed to chlorpyrifos, and the ability of oximes to reactivate the phosphorylated ChE activity. Results indicated that these esterase activities are a suitable multibiomarker scheme for monitoring OP exposure due to their high sensitivity to OP inhibition and slow recovery to full activity levels following pesticide exposure. Moreover, oximes reactivated the inhibited ChE activity of the earthworms exposed to 12 and 48 mg kg−1 chlorpyrifos during the first week following pesticide exposure. This methodology is useful for providing evidence for OP-mediated ChE inhibition in individuals with a short history of OP exposure (≤1 week); resulting a valuable approach for assessing multiple OP exposure episodes in the field. Esterase inhibition combined with oxime reactivation methods is a suitable approach for monitoring organophosphate contamination

International audience | BioROC aims to improve knowledge of the biofilm and soil macrofauna influence in the water reuse field including constructed wetland (CW) and irrigation. This project will contribute to the adoption of innovative practices, working in two living labs of Toulouse Métropole and Montpellier Méditerranée Métropole. The challenge is to test a panel of CW with a gradient of biodiversity (+/- plants and +/- invertebrate addition), to identify the most suited modalities for reuse in irrigation. BioROC aims to find the best match between irrigation devices and CW functioning. The obstacles to be overcome are the clogging of filters and drip-irrigation systems by biofilm. The quality of the biofilms and their dynamics will be described using metagenomic analyses, sensors, and modeling. These results obtained in an interdisciplinary approach make it possible to improve current capacities to control biofilm in drip-irrigation systems and the accuracy of bioinspired CW for water reuse.

Although microplastics (MPs) are ubiquitous in agricultural soil, little is known about the effects of MPs combined with pesticides on soil organisms and their biogenic transport through the soil profile. In this study, we conducted mesocosm experiments to observe the effects of microplastics (polyethylene (LDPE-MPs) and biodegradable microplastics (Bio-MPs)) and chlorpyrifos (CPF) on earthworm (Lumbricus terrestris) mortality, growth and reproduction, as well as the biogenic transport of these contaminants through earthworm burrows. The results showed that earthworm reproduction was not affected by any treatment, but earthworm weight was reduced by 17.6% and the mortality increased by 62.5% in treatments with 28% Bio-MPs. Treatments with 28% LDPE-MPs and 7% Bio-MPs combined with CPF showed greater toxicity while the treatment with 28% Bio-MPs combined with CPF showed less toxicity on earthworm growth as compared to treatments with only MPs. The treatments with 1250 g ha−1 CPF and 28% Bio-MPs significantly decreased the bioaccumulation of CPF in earthworm bodies (1.1 ± 0.2%, w w−1), compared to the treatment with CPF alone (1.7 ± 0.4%). With CPF addition, more LDPE-MPs (8%) were transported into earthworm burrows and the distribution rate of LDPE-MPs in deeper soil was increased. No effect was observed on the transport of Bio-MPs. More CPF was transported into soil in the treatments with LDPE-MPs and Bio-MPs, 5% and 10% of added CPF, respectively. In addition, a lower level of the CPF metabolite 3,5,6-trichloropyridinol was detected in soil samples from the treatments with MPs additions than without MP additions, indicating that the presence of MPs inhibited CPF degradation. In conclusion, Bio-MPs caused significant toxicity effects on earthworms and the different types of MPs combined with CPF affected earthworms differently, and their transport along the soil profile. Thus, further research is urgently needed to understand the environmental risks of MPs and MP-associated compounds in the soil ecosystem.

As a result of the efforts to introduce the principle of the 3Rs (replacement, reduction, and refinement) into animal testing, alternative in vitro skin corrosion test methods have been developed and standardized globally. However, alternative in vitro skin corrosion test methods have some limitations in terms of the use of humanely killed rats or commercial models and kits. The present study focused on the applicability of invertebrates as alternative in vivo skin models. Even though earthworm skin comprises the same biomolecules as human skin, the possibility of using earthworm skin as an alternative for skin testing remains unexplored. In this study, we developed a half-pipe tool for earthworm skin corrosion testing and optimized the test protocol. Subsequently, the applicability of the earthworm half-pipe assay for corrosion testing with six chemicals, including inorganic acids, organic acids, and alkalis, was investigated using stereomicroscopy and electron microscopy. It was observed that the specific concentrations for earthworm skin corrosion were lower than those for animal or in vitro tests. Therefore, the sensitivity of the earthworm half-pipe assay indicates that it could be useful as a screening tool before conducting in vivo animal tests or in vitro skin tests. This new method can contribute to research on alternative skin corrosion tests by reducing ethical issues, time, and cost while achieving effective results.

Rare earth elements inevitably release into the soil due to their widespread application. However, it is unclear how they affect the soil animals. The study surveyed the growth and physiological responses of earthworm (Eisenia fetida) exposed into artificial soils spiked with La, Ce, and their mixture, and actual mine soil collected from an abandoned La–Ce mining area (Mianning, Sichuan). The results showed that the 1000–1200 mg/kg combined exposure in two soils induced significant histopathological and phenotypic changes of earthworms. Concentration significantly affected the superoxide dismutase (SOD), peroxidase (POD), malondialdehyde (MDA), and protein of E. fetida and the effects differentiated with the prolonging duration. These indicators were negatively affected under the La stress ≥800 mg/kg (SOD, POD, and protein), the 1200 mg/kg (SOD), Ce stress ≥1000 mg/kg (protein), and the combination ≥800 mg/kg (SOD, POD) and ≥1000 mg/kg (protein). Artificial combination had −15.04% (SOD), 8.87% (POD), 5.64% (MDA), and −8.34% (protein) difference compared with the contamination soil, respectively. Overall, E. fetida respond sensitively under the La and Ce stress, the antioxidant defense system and the lipid peroxidation were stimulated, and the artificial soil might overestimate eco-toxicological effect.

The scale of variation in species sensitivity to toxicants has been theoretically linked to mode of action. Specifically, it has been proposed there will be greater variations for chemicals with a putative specific biological target than for toxicants with a non-specific narcotic mechanism. Here we test the hypothesis that mode of action is related to variation in sensitivity in a specifically designed experiment for species from a single ecologically important terrestrial taxa, namely earthworms. Earthworm toxicity tests were conducted with five species for four chemicals, providing a series of increasingly complex modes of action: a putative narcotic polycyclic aromatic hydrocarbon (fluoranthene), and three insecticides (chlorpyrifos, cypermethrin, imidacloprid) with known neuronal receptor targets. Across all the chemicals, the standard epigeic test species Eisenia fetida and Lumbricus rubellus, were generally among the two least sensitive, while the endogenic Aporrectodea caliginosa and Megascolecidae Amynthas gracilis were generally more sensitive (never being among the two least sensitive species). This indicates a potential for bias in the earthworm ecotoxicology literature, which is dominated by studies in epigeic Lumbricidae, but contains few endogeic or Megascolecidae data. Results confirmed the lowest range of variation in sensitivities for effects on reproduction was for fluoranthene (2.5 fold). All insecticides showed greater variation for species sensitivity (cypermethrin: 7.5 fold, chlorpyrifos: 10.3 fold, imidacloprid: 31.5 fold) consistent with the specific mechanisms of the pesticides. Difference in toxicodynamics, based on mode of action specificity and receptor complexity was reflected in the magnitude of sensitivity variation. However, measurements of tissue concentrations also indicated the potential importance of toxicokinetics in explaining species sensitivity variations for chlorpyrifos and cypermethrin.

Microplastics (MPs) are plastic fragments less than 5 mm, which may have adverse impacts on organisms. In this study, we investigated the impacts and mechanisms of polystyrene MPs (10 μm and 100 μm) and nanoplastics (NPs, 100 nm) with different concentrations (10 mg/kg and 100 mg/kg) in soil on the uptake of metal Cd and semi-metal As in earthworms, Eisenia fetida. MPs facilitated the accumulation of (semi-)metals via damaging the integrity of earthworm intestine, and earthworms accumulated more (semi-)metals in MP treatment groups than NP treatment groups, especially in group of 100 mg/kg of 10 μm MP with concentrations of 1.13 mg/kg and 32.7 mg/kg of Cd and As, respectively. Higher genotoxicity to earthworms was observed for MPs than NPs. Antioxidant enzymes activity and their mRNA gene relative expression levels indicated that MPs with high concentration induced severer damage to earthworms, thus resulting in the increased accumulation of (semi-)metals by earthworms. In addition, proteomic and metabolomic analysis revealed that MPs (100 ppm of 10 μm) disturbed the earthworm immune and metabolic systems, resulting in the highest accumulation of (semi-)metals in earthworms. This study clarifies the influence mechanisms of MPs with different sizes and levels on the accumulation of (semi-)metals by terrestrial invertebrates.

A long-term case of residual pollution is studied after 20 years since the largest mining accident in Spain (the Aznalcóllar spill) happened. This pollution is manifested through a surface zoning consisting of bare soils (B0), sparsely vegetated soils (B1), and densely vegetated and recovered soils (B2). A biopiles treatment with a mixture of contaminated soils (B0 and B1) with recovered soils (B2) at 50% (w/w), and vermicompost addition (50 tons ha-1) was evaluated. To assess the effectiveness of treatments, total, water-soluble, and bioavailable fractions of the most polluting elements in the zone (Cu, Zn, As, Pb, Cd, and Sb) was analyzed. To evaluate the potential risk of contamination for the ecosystem, a bioassay with earthworm Eisenia andrei was carried out. Twenty years after the accident, there are still soils where total As and Pb exceed the regulatory levels and water-soluble Zn and As exceed the toxicity guidelines. According to toxicity bioassay, weight variation and juvenile production of earthworms showed an improvement after biopiles treatment, with values that trend to be similar to those of recovered soils. The only bioaccumulated element in earthworms was Cd (BAF>1), both in polluted as in treated soils, which indicates the possible existence of exclusion mechanisms of the other pollutants by earthworms. The comparison between biopiles and polluted soils showed no significant differences for the bioaccumulation factor of trace elements, with the exception of Zn and Cu, which slightly increased after treatment. According to our results, biopiles treatment combined with vermicompost addition is a good technique for the recovery of residual contaminated areas, by the improvement of soil properties and the reduction of the potential toxicity; anyway, monitoring of soils and organisms is needed to prevent the increase of bioavailability of some potentially pollutant elements over time.

Concentrations as high as thousands of milligrams per kilogram (dry weight) of nonylphenol (NP), an endocrine-disrupting chemical of great concern, have been reported in soil. Soil is considered one of the primary pathways for exposure of crop plants to NP. However, there have been few studies on the toxicity of soil NP to crop plants, especially with comprehensive consideration of the application of organic fertiliser which is a common agricultural practice. In this study, tomato plants were grown in soils treated with NP in the presence and/or absence of earthworm casts (EWCs). After four weeks, we tested the physiological and biochemical responses (accumulative levels of hydrogen peroxide (H₂O₂) and superoxide anion radicals (O₂-·), total chlorophyll content, degree of membrane lipid peroxidation, activities of defence-related enzymes, and level of DNA damage) and the changes in plant growth (elongation and biomass). The growth inhibition, reactive oxygen species (H₂O₂ and O₂-·) accumulation, decrease in chlorophyll content, increase in activity of defence-related enzymes (including superoxide dismutase, peroxidase, catalase, ascorbate peroxidase, glutathione S-transferase and glutathione reductase), enhancement of membrane lipid peroxidation, and DNA damage in NP-treated seedlings were clearly reversed by the intervention of EWCs. In particular, the suppressed elongation, biomass, and chlorophyll content in tomato plants exposed to NP alone were significantly restored by EWCs to even greater levels than those of the undisturbed control. In other words, EWCs could efficiently invigorate the photosynthesis of crops via up-regulating the chlorophyll content, thereby overwhelming the NP stress on plant growth. Accordingly, except for reducing the bioavailability of soil NP as reported in our previous study, EWCs could also help crop plants to cope with NP stress by strengthening their stress resistance ability. Our findings are of practical significance for the formulation of strategies to relieve the negative effects of soil NP on crop growth.