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.

Nitrous oxide (N₂O) is a devastating greenhouse gas and acts as an ozone-depleting agent. Earthworms are a potential source of soil N₂O emissions. Application of biochar can mitigate earthworm-induced N₂O emissions. However, the underlying interactive mechanism between earthworms and biochar in soil N₂O emissions is still unclear. A 35-day laboratory experiment was conducted to examine the soil N₂O emission dynamics for four different treatments, earthworm presence with biochar application (EC), earthworm presence without biochar application (E), earthworm absence with biochar application (C) and earthworm absence without biochar application, and the control. Results indicated a negative impact of biochar on earthworm activity, displaying a significantly (p ≤ 0.05) lower survival rate and biomass of earthworms in treatment EC than E. Compared with the control, earthworm presence significantly (p ≤ 0.05) increased cumulative N₂O emissions, while application of biochar in the presence of earthworms significantly (p ≤ 0.05) decreased cumulative N₂O emissions (485 and 690 μg kg⁻¹ for treatments EC and E, respectively). Treatments E and EC significantly (p ≤ 0.05) increased soil microbial biomass carbon (MBC), ammonium (NH₄⁺-N), nitrate (NO₃⁻N), and dissolved organic carbon (DOC) content and soil pH as compared with the control. The gene copy number of 16 S rRNA, AOA, AOB, nirS, and nosZ increased for all treatments when compared with the control; however, a significant (p ≤ 0.05) difference among the studied genes was only observed for the nosZ gene (2.05 and 2.56 × 10⁶ gene copies g⁻¹ soil for treatments E and EC, respectively). Earthworm-induced soil N₂O emissions were significantly (p ≤ 0.05) reduced by biochar addition. The possible underlying mechanisms may include: (1) short-term negative impacts on earthworm activity; (2) a change of functional gene abundance in earthworm casts; and (3) an increase in soil pH due to addition of biochar.

Earthworms and arbuscular mycorrhizal fungi (AMF) act synergistically in the rhizosphere and may increase host plant tolerance to Cd. However, mechanisms by which earthworm-AMF-plant partnerships counteract Cd phytotoxicity are unknown. Thus, we evaluated individual and interactive effects of these soil organisms on photosynthesis, antioxidant capacity, and essential nutrient uptake by Solanum nigrum, as well as on soil quality following Cd exposure (0–120 mg kg⁻¹). Decreases in biomass and photosynthetic activity, as well as nutrient imbalances were observed in Cd-stressed plants; however, the addition of AMF and earthworms reversed these effects. Cd exposure increased superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities, whereas inoculation with Rhizophagus intraradices decreased those. Soil enzymatic activity decreased by 15–60% with increasing Cd concentrations. However, Cd-mediated toxicity was partially reversed by soil organisms. Earthworms and AMF ameliorated soil quality based on soil enzyme activity. At 120 mg kg⁻¹ Cd, the urease, catalase, and acid phosphatase activities were 1.6-, 1.4-, and 1.2-fold higher, respectively, in soils co-incubated with earthworms and AMF than in uninoculated soil. Cd inhibited shoot Fe and Ca phytoaccumulation, whereas AMF and earthworms normalized the status of essential elements in plants. Cd detoxification by earthworm-AMF-S. nigrum symbiosis was manifested by increases in plant biomass accumulation (22–117%), chlorophyll content (17–63%), antioxidant levels (SOD 10–18%, POD 9–25%, total polyphenols 17–22%, flavonoids 15–29%, and glutathione 7–61%). It also ameliorated the photosynthetic capacity, and macro- and micronutrient statuses of plants; markedly reduced the levels of malondialdehyde (20–27%), superoxide anion (29–36%), and hydrogen peroxide (19–30%); and upregulated the transcription level of FeSOD. Thus, the combined action of earthworms and AMF feasibly enhances metal tolerance of hyperaccumulating plants and improves the quality of polluted soil.

Toxicokinetic (TK) model provides a new approach to mechanistically elucidate the natural variation of metal handling strategy by adaptive and sensitive earthworm populations. Here, TK model was applied to explore the metal handling and resistance strategy of wild Metaphire californica with different historical exposure history through a 12-day re-exposure and another 12-day elimination incubation. M. californica populations showed different kinetic strategies for non-essential metals (Cd and Pb) and essential metals (Zn and Cu), which were closely related to their exposure history. M. californica from the most serious Cd-contaminated soil showed the fastest kinetic rates of both Cd uptake (K₁ = 0.78 gₛₒᵢₗ/gwₒᵣₘ/day) and elimination (K₂ = 0.23 day⁻¹), and also had the lowest Cd half-life (t₁/₂ = 3.01 day), which demonstrated the potential Cd-resistance of wild M. californica from Cd-contaminated soils. Besides, the comparative experiment showed totally different metal kinetics of laboratory Eisenia fetida from field M. californica, suggesting the impacts of distinct exposure history and species-specifical sensitivities. These findings provide a novel approach to identify and quantify resistance using TK model and also imply the risk of overlooking existing exposure background and interspecies extrapolation in eco-toxicological studies and risk assessments.

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.

The large accumulation of heavy metals in the soil surrounding steel factories has become a severe environmental problem. However, few studies have focused on how the earthworm gut microbiota responds to heavy metals in the soil. This study used research sites at a steel factory in Nanjing, China, to investigate how the soil bacterial community and earthworm gut microbiota respond differently to heavy metal contamination using Illumina high-throughput sequencing targeting 16S rRNA genes. The bacterial community of earthworm guts showed a distinct structure compared with that of the soil, featuring a higher relative abundance of Proteobacteria (45.7%) and Bacteroidetes (18.8%). The bacterial community in the earthworm gut appeared more susceptible to heavy metal contamination compared with the soil community. For example, we identified 38 OTUs (Operational taxonomic units) significantly influenced by contamination among 186 abundant OTUs in the soil, whereas 63 out of the 127 abundant OTUs in the earthworm gut were altered significantly under contamination. This susceptibility may be partly explained by the lower alpha diversity and distinct microbial interactions in the gut. In addition, the accumulation of heavy metals also stimulated the growth of potential plant growth promoting bacteria (PGPB) in the earthworm gut, especially those related to indole-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) production, which may potentially benefit the phyto-remediation of heavy metals. These results contribute to our understanding of the soil biota and its interactions under heavy metal contamination and may provide further insights into the phyto-remediation of metal-contaminated soil.

In this study, we explored the influence of two metal oxide nanoparticles, nano CuO and nano ZnO (10, 50, 250 mg/kg), on accumulation of bifenthrin (100 μg/kg) in earthworms (Eisenia fetida) and its mechanism. The concentrations of bifenthrin in earthworms from binary exposure groups (bifenthrin + CuO and bifenthrin + ZnO) reached up to 23.2 and 28.9 μg/g, which were 2.65 and 3.32 times of that in bifenthrin exposure group without nanoparticles, respectively, indicating that nanoparticles facilitated the uptake of bifenthrin in earthworms. The contents of biomarkers (ROS, SOD, and MDA) in earthworms indicated that nanoparticles and bifenthrin caused damage to earthworms. Ex vivo test was utilized to investigate the toxic effects of the pollutants to cell membrane of earthworm coelomocytes and mechanism of increased bifenthrin accumulation. In ex vivo test, cell viability in binary exposure groups declined up to 30% and 21% compared to the control group after 24 h incubation, suggesting that coelomocyte membrane was injured by the pollutants. We conclude that nanoparticles damage the body cavity of earthworms, and thus lead to more accumulation of bifenthrin in earthworms. Our findings provide insights into the interactive accumulation and toxicity of nanoparticles and pesticides to soil organisms.