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

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.

Fluoxastrobin is one of the most widely used strobilurin fungicides, however, application of the fungicides may result in soil residues leading to environmental damage including oxidative stress and damage to sentinel organisms (i.e. earthworms). While this has been demonstrated in artificial soil, the biochemical response of Eisenia fetida exposed to fluoxastrobin in natural soils is unclear. This study utilized three typical natural soils (fluvo-aquic soils, red clay, and black soils) to evaluate the biochemical response of Eisenia fetida exposed to fluoxastrobin (0.1, 1.0, 2.5 mg/kg) including the production of reactive oxygen species, impact on three enzyme activities, lipid peroxidation, and 8-hydroxydeoxyguanosine after a 4-week exposure. The effects of fluoxastrobin on Eisenia fetida in different soils were assessed using an integrated biomarker response (IBR). The findings may be possible to state that the toxic effects of fluoxastrobin in artificial cannot exactly represent that in natural soils. Specifically, the fluoxastrobin subchronic toxicity was highest in red clay and lowest in black soil among the three natural soils. Furthermore, the 8-OHdG content was more sensitive to fluoxastrobin in all six environmental indicators of the present study.

Present study aims to examine the efficiency of co-vermicomposting amended with activated sludge and E. fetida earthworm for bioremediation of diesel and gasoline from contaminated soil. The diesel and gasoline removal efficiency and degradation rates coefficients were estimated with gas chromatography (GC) analysis and first-order kinetics. The removal of gasoline and diesel in different co-vermicomposting processes with and without E. fetida ranged between 65-100% and 24.94–63.93%, respectively within 90- day experiment. Removal of gasoline and diesel increased in soil with addition of earthworm (E. fetida); higher degradation rate coefficients (k) were observed for co-vermicomposting with earthworm compared with co-vermicomposting processes. The highest k (0.014) for diesel degradation was estimated for microcosm reactor 4 (R4), where high numbers of E. fetida accelerate the less biodegradable organic contaminant from the soil matrices. The reasonable survival rates of earthworms in exposure to high concentration of petroleum-derivatives contaminated soils indicated increased activity of ligninolytic diesel–degrading earthworms and microorganisms. Therefore, co-vermicomposting amended with activated sludge is suggested as feasible and promising technologies for bioremediation of high content of organic contaminants from the soil matrices.

The growing use of graphene-based nanomaterials (GBNs) for various applications increases the probability of their environmental releases and calls for a systematic assessment of their potential impacts on soil invertebrates that serve as an important link along terrestrial food chains. Here, we investigated the response of earthworms (Eisenia fetida) to three types of multi-layer graphenes (MLGs) (G1, G2 and G3 with 12–15 layers) with variable morphology (lateral sizes: 7.4 ± 0.3, 6.4 ± 0.1 and 2.8 ± 0.1 μm; thicknesses: 5.0 ± 0.1, 4.2 ± 0.1 and 4.0 ± 0.2 nm, respectively) and hydrophobicity ((O + N)/C ratios: 0.029, 0.044 and 0.075; contact angles: 122.8, 118.8 and 115.1°, respectively). Exposure to these materials was conducted for 28 days (except for 48-h avoidance test) separately in potting or farm soil at 0.2% and 1% by weight. Earthworms avoided both soils when amended with 1% of the smaller and more hydrophilic MLGs (G2 and G3), leading to a decreased trend in worm cocoon formation. The smallest and most hydrophilic MLG (G3), which was easier to assimilate, also significantly inhibited the viability (20.2–56.0%) and mitochondrial membrane potential (32.0–48.5%) of worm coelomocytes in both soils. In contrast, oxidative damage (indicated by lipid peroxides) was more pronounced upon exposure to more hydrophobic and larger graphenic materials (G1 and G2), which were attributed to facilitated adhesion to and disruption of worm membranes. These findings highlight the importance of MLG morphology and hydrophobicity in their potential toxicity and mode of action, as well as ecological risks associated with incidental and accidental releases.