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.

The enantioselective toxic effect and environmental behavior of chiral pesticides have attracted increasing research attention. In this study, the enantioselective toxicity and residues of hexaconazole (HEX) in earthworms (Eisenia fetida) were investigated. In the present study, significant enantioselective degradation characteristics were observed in artificial soil with the R-enantiomer preferentially degrading (p < 0.05); however, no significant enantioselective bioaccumulation was observed in the earthworms (p > 0.05). The acute toxicity of S-(+)-HEX was higher than that of R-(−)-HEX in earthworms, with 48-h LC₅₀ values of 8.62 and 22.35 μg/cm², respectively. At 25 mg/kg, enantiospecific induction of oxidative stress was observed in earthworms; moreover, S-(+)-HEX had a greater influence on the contents of malonaldehyde, cytochrome P450, and 8-hydroxy-2-deoxyguanosine than R-(−)-HEX. These results were consistent with those of the enrichment analysis of differentially expressed genes. The transcriptome sequencing results showed that S-(+)-HEX had a more significant influence on steroid biosynthesis, arachidonic acid metabolism, and cell cycle processes than R-(−)-HEX, leading to abnormal biological function activities. These results indicate that S-(+)-HEX may pose a higher risk to soil organisms than R-(−)-HEX. This study suggests that the environmental risk of chiral pesticides to nontarget organisms should be assessed at the enantiomeric level.

Selenium (Se) is an essential micronutrient for animals with a narrow margin between essentiality and toxicity. Se toxicity is largely related to inorganic forms of Se in soil, i.e., selenite and selenate that enter food chains through plant uptake, threatening higher trophic level organisms. This experiment investigated effects of earthworm activity on Se bioavailability in soil and the subsequent plant uptake, using earthworm Eisenia fetida and bean plant Phaseolus vulgaris L, both exposed to either selenite or selenate at 1 or 4 mg Se kg⁻¹ for 16 weeks. Plants took up selenate (up to 221-fold) faster than selenite, with up to 84% of the Se rapidly transported to shoots. In the presence of earthworms, Se accumulation obviously increased for selenate-supplied plants, leading to an up to 4% increase in Se translocation factor for all treatments except for 1 mg kg⁻¹ selenite treatment. Earthworms also concentrated Se faster in tissues (up to 274 mg kg⁻¹ DW) at exposure to selenate. For Se toxicity, Se speciation analysis was conducted on the plants and earthworms using XAS. Compared to worm-free treatments, the percentage of organo-Se, i.e., SeMet and CysSeSeCys, increased in beans (up to 34%) in the presence of earthworms for selenate, while the elemental Se portion was significantly reduced or absent, opposite to the results for selenite. Surprisingly, elemental Se (up to 65%) dominated earthworms, regardless of the form of Se supplied. In conclusion, earthworms clearly enhanced Se uptake and translocation in plants, leading to elevated Se levels in shoots. To prevent resulting hazards to humans and other animals, caution should be taken while consuming the shoots, particularly beans, harvested from the Se contaminated soil where earthworm activity is high. Finally, the significant reduction in soil Se suggests phytoextraction of Se from the soil could be improved using earthworms as an aid to plants.

The remediation of cadmium (Cd) contaminated soil has become a global problem due to its toxicity to living organisms. In this study, earthworm (Eisenia fetida) alone or combined with EDTA or bean dregs were used for Cd removal from soils. The total and available Cd in soils, soil physicochemical and biological (soil enzyme) properties, Cd accumulation in the earthworm and its antioxidant responses towards Cd, were determined during the 35 days of soil incubation experiment. Our results showed that earthworms were capable of removing Cd from soils, and the remediation process was accelerated by both EDTA and bean dregs. By translocation of Cd from soils, the content of Cd in earthworm steadily increased with the exposure time to 8.11, 12.80, and 9.26 mg kg⁻¹ on day 35 for T2 (earthworm alone), T3 (EDTA enhancement), and T4 (bean dregs enhancement), respectively. Consequently, a great reduction in the Cd contents in soils was achieved in T3 (36.53%) and T4 (30.8%) compared with T2 (28.95%). The concentrations of water/DTPA extractable Cd were also reduced, indicating the low Cd mobility after amendment. Finally, the soil became more fertile and active after wermi-remediation. The soil pH, EC, NO₃⁻-N, available P, and K contents increased, while soil SOM, DOC, and NH₄⁺-N contents were decreased. There were higher soil enzyme activities (including acid phosphatase activity, β-glucosidase activity, and urease activity) among treatments with earthworms. Additionally, the operational taxonomic units (OTUs) increased by 100–150 units, and the higher chao1 and Shannon indexes indicated the enhanced microbial community after wermi-remediation, especially among treatment with EDTA and bean dregs. Therefore, we concluded that earthworms, alone or combined with EDTA and bean dregs, are feasible for the remediation of Cd-contaminated soil.

Humic acid (HA) plays vital roles in regulating the environmental behaviors of metals and thus their toxicity to biota. However, the inner relation between metal bioavailability to soil organisms and the presence of HA with different molecular weight (Mw) is not well documented. In this study, we separated HAs into four fractions with Mw range of 5-30k Da, and discussed their ability to alleviating the toxicity of Cd and Pb to earthworm. The bioaccumulation capacities (Cₘₐₓ) increased in order of: UF1<UF2<UF3<UF4, which is in line with the variations of bioavailable concentrations of Cd and Pb in soil. Variations of Mw and binding capacities of HA determine the accumulation behavior in soil solution. The unsatisfactory of biotic ligand model fitting and the differences in fractions of the total biotic ligand sites (f) in earthworm bound by Cd and Pb suggested that only free species of Cd could be considered as biological available to earthworm, while the Pb–HAs complexes have potential ability to interact with earthworm membrane. Antioxidant enzymes are effective biomarkers, and HA with lower Mw play more important roles in restricting the toxicity of soil Cd and Pb to earthworm. These results reveal the different mechanism for HA controlling metal bioavailability between Cd and Pb in soil environment.

Environmental pollution can disrupt the interactions between animals and their symbiotic bacteria, which can lead to adverse effects on the host even in the absence of direct chemical toxicity. It is therefore crucial to understand how environmental pollutants affect animal microbiomes, especially for those chemicals that are designed to target microbes. Here, we study the effects of two biocidal nanoparticles (NPs) (Ag and CuO) on the soil bacterial community and the resident gut microbiome of the earthworm Eisenia fetida over a 28-day period using metabarcoding techniques. Exposures to NPs were conducted following OECD test guidelines and effects on earthworm reproduction and juvenile biomass were additionally recorded in order to compare effects on the host to effects on microbiomes. By employing a full concentration series, we were able to link pollutants to microbiome effects in high resolution. Multivariate analysis, differential abundance analysis and species sensitivity distribution analysis showed that Ag-NPs are more toxic to soil bacteria than CuO-NPs. In contrast to the strong effects of CuO-NPs and Ag-NPs on the soil bacterial community, the earthworm gut microbiome is largely resilient to exposure to biocidal NPs. Despite this buffering effect, CuO-NPs did negatively affect the relative abundance of some earthworm symbionts, including ‘Candidatus Lumbricincola’. Changes in the soil bacterial community and the earthworm microbiome occur at total copper concentrations often found or modelled to occur in agricultural fields, demonstrating that soil bacterial communities and individual taxa in the earthworm microbiome may be at risk from environmental copper exposure including in nanomaterial form.

Degradation of metolachlor in surface soil is extremely important to its potential mobility and overall persistence. In this study, the effects of earthworms (Eisenia fetida) on the degradation of metolachlor at two concentration levels (5 and 20 mg kg⁻¹) in soil were investigated via the column experiment. The degradation kinetics of metolachlor indicate that addition of earthworms enhances metolachlor degradation significantly (P < 0.05), with the enhanced degradation rate of 30% and 63% in the low and high concentration treatments at the 15th day, respectively. Fungi rather than bacteria are primarily responsible for metolachlor degradation in soil, and earthworms stimulate metolachlor degradation mainly by stimulating the metolachlor-degrading functional microorganisms and improving fungal community structure. Earthworms prefer to promote the possible fungal degraders like order Sordariales, Microascales, Hypocreales and Mortierellales and the possible bacteria genus Rubritalea and strengthen the relationships between these primary fungi. Two metabolites metolachlor oxanilic (MOXA) and moetolachlor ethanesulfonic acid (MESA) are detected in soil and earthworms in the high concentration treatments. Earthworms stimulate the formation of MOXA and yet inhibit the formation of MESA in soil. Another metabolite metolachlor-2-hydroxy (M2H) is also detected in earthworms, which is reported firstly. The study provides an important information for the remediation of metolachlor-polluted soil.

Although, health and environmental hazards of Ni are ironclad; however, that of Nickle oxide nanoparticles (NiO-NPs) are still obscure. Therefore, impact of NiO-NPs exposure (0, 5, 50, 200, 500 and 1000 mg kg⁻¹ soil) on the earthworm (Eisenia fetida) survival (at 28th day), reproduction (at 56th day), histopathology, ultrastructures, antioxidant enzymes and oxidative DNA damage was appraised in full life cycle study. Lower concentrations of NiO-NPs (5, 50 and 200) did not influence the survival, reproduction and growth rate of adult worms significantly. However, reproduction reduced by 40–50% with 500 and 1000 mg kg⁻¹ exposure, which also induced oxidative stress leading to DNA damage in earthworms. Ultrastructural observation and histology of earthworms exposed to higher NiO-NPs concentrations revealed abnormalities in epithelium layer, microvilli and mitochondria with underlying pathologies of epidermis and muscles, as well as adverse effects on the gut barrier. To the best of our knowledge, this is the first study unveiling the adverse effects of NiO-NPs on a soil invertebrate (Eisenia fetida). Our findings clue towards looking extensively into the risks of NiO-NPs on soil organisms bearing agricultural and environmental significance.

Hexabromocyclododecanes (HBCDDs) are common chemical additives in expanded polystyrene foam (EPS). To evaluate the bioaccumulation potential of endogenous HBCDDs in EPS microparticles by earthworms, two ecologically different species of earthworms (Eisenia fetida and Metaphire guillelmi) were exposed to soil added with EPS microparticles of different particle sizes (EPS2000, 830–2000 μm and EPS830, <830 μm). To clarify the accumulation mechanisms, leaching experiments using EPS microparticles in different solutions were conducted. After exposure to EPS microparticles-amended soils (S-EPS) for 28 d, the total concentrations of HBCDDs reached 307–371 ng g−1 dw in E. fetida and 90–133 ng g−1 dw in M. guillelmi, which were higher than those in earthworms exposed to the soil that was artificially contaminated with a similar level of HBCDDs directly (ACS). The accumulation of HBCDDs in earthworms was significantly influenced by EPS microparticles' size and earthworms' species. The total concentrations of HBCDDs in earthworms' cast were significantly higher than the theoretical concentration of HBCDDs in S-EPS, which suggested that EPS microparticles can be ingested by earthworms. The release rate of HBCDDs from EPS5000 (2000–5000 μm) into water-based solutions (<1%) after a 3.5-h incubation was far lower than that into earthworm digestive fluid (7%). These results illustrated that the ingestion of EPS microparticles and consequent solubilization of HBCDDs by digestive fluid play an important role in the accumulation of HBCDDs contained in EPS microparticles in earthworms. After a 28-d incubation with the soil solution, 4.9% of the HBCDDs was accumulatively leached from the EPS5000, which indicated that HBCDDs can be released from EPS microparticles to soil environment, and then accumulated by earthworms. Moreover, similar to those exposed to ACS, the diastereoisomer- and enantiomer-specific accumulation of HBCDDs in earthworms occurred when exposed to S-EPS. This study provides more evidence for the risk of microplastics to the soil ecosystem.