Occurrence and Fate of Fluoroalkyl Sulfonamide-Based Copolymers in Earthworms-Bioavailability, Transformation, and Potential Impact of Sludge Application.

To date, considerable knowledge and data gaps regarding the occurrence, environmental levels, and fate of polymeric perfluoroalkyl and polyfluoroalkyl substances (PFAS) exist. In the present study availability, accumulation, and transformation of C4- and C8-fluoroalkylsulfonamide (FASA)-based copolymers were assessed in laboratory-grown earthworms (Eisenia fetida, triplicate of exposure tests and control). Further, a field study on earthworms (18 pooled samples) in sludge-amended soil was conducted to assess the environmental impact of sludge-amended soil with regard to the FASA-based copolymers, together with the applied sludge (n = 3), and the field soils during the period (n = 4). In the laboratory study, the FASA-based copolymers were taken up by the earthworms in concentrations between 19 and 33 ng/g of dw for the C8- and between 767 and 1735 ng/g of dw for the C4-FASA-based copolymer. Higher biota soil accumulation factors (BAFs) were observed for the copolymer with a longer perfluorinated side-chain length (C8, average BAF value of 0.7) compared to the copolymer with a shorter side-chain length (C4, average BAF value of 0.02). Perfluorooctane sulfonamidoacetates (FOSAAs) and perfluorooctane sulfonamide (FOSA), including both branched and linear isomers, were detected after exposure to the C8-FASA-based copolymer. Two metabolites were detected in the earthworms exposed to the C4-FASA-based copolymer: perfluorobutanesulfonamide (FBSA) and perfluorobutanesulfonic acid (PFBS). Although the presence of other monomers or impurities in the copolymer formulation cannot be ruled out, the present laboratory study suggests that the FASA-based copolymers may be an indirect source of lower molecular weight PFAS in the environment through transformation. Elevated levels of C8-FASA-based copolymer were found in the field sludge-amended soil compared to nontreated soil (32 versus 11 ng/g d.w.), and higher concentrations of PFAS in earthworms living in sludge-amended soil compared to nontreated soil (566 versus 103 ng/g d.w.) were observed. These findings imply that the application of sludge is a potential pathway of PFAS to the environment.

Effects of earthworm and arbuscular mycorrhizal fungal inoculation on carbon component accumulation and allocation in rocky desertification soils.

Exploring the responses of carbon component accumulation and allocation to arbuscular mycocorrhizal fungi (AM) and earthworm inoculation can provide reference for improving carbon sequestration potential and bioremediation efficiency in rocky desertification soils. In this study, we chose Fraxinus malacophylla as the host plant to inoculate with Funneliformis mosseae (FM), earthworm (E), and E+FM, using no earthworm and mycorrhizae addition as CK to examine the spatiotemporal variations in soil carbon components (i.e., total organic carbon, microbial biomass carbon, easily oxidized organic carbon, and recalcitrant organic carbon) and their allocation (i.e., microbial biomass carbon/total organic carbon, easily oxidized organic carbon/total organic carbon, and recalcitrant organic carbon/total organic carbon). The results showed that 1) The respective and interactive inoculation of E and AM significantly promoted the accumulation of each carbon component. In contrast with the control, the average carbon component levels under three inoculation treatments were ranked as E+FM>E>FM. The three inoculation treatments significantly promoted soil microbial carbon/total organic carbon (30.5%-68.5%) and easily oxidized carbon/total organic carbon (31.2%-39.2%), but decreased recalcitrant organic carbon/total organic carbon (2.9%-16.2%). 2) The spatiotemporal variation in accumulation and allocation of soil carbon components varied between the inoculation treatments. The maximum value of each carbon component occurred in June. The increase in each carbon component was significantly higher in E+FM (33.0%-122.1%) than that in E (31.2%-95.4%) and FM (9.2%-41.3%). The maximum value of microbial biomass carbon/total organic carbon and easily oxidized organic carbon/total organic carbon was observed in June, while that of recalcitrant organic carbon/total organic carbon was recorded in December. In contrast with CK, the amplitude of variation in the proportion of carbon components in total organic carbon under the three inoculation treatments was ranked as E+FM>E>FM. The accumulation and allocation of all carbon components decreased (9.7%-146.2%) along the soil profile. The level of carbon components in the E treatment decreased the smallest. The microbial biomass carbon/total organic carbon and easily oxidized carbon/total organic carbon decreased the least and the recalcitrant organic carbon/total organic carbon decreased the greatest under the E+FM treatment. 3) Changes in soil physicochemical properties under the three inoculation treatments significantly affected the accumulation and allocation of organic carbon components. Soil pH was negatively correlated with carbon component accumulation and allocation, whereas other soil variables were positively correlated with them. 4) The results of principal component analysis showed that soil water content, total nitrogen, and total phosphorus were the main factors driving carbon component accumulation, while soil water content, total phosphorus, and pH were the main factors controlling carbon component allocation. Therefore, we concluded that the earthworms, AM fungi and their interaction affected the accumulation and allocation of carbon components in Yunnan rocky desertification soils, which would primarily depend on the changes of soil water content, acid-base property, as well as nitrogen and phosphorus conditions.

Leveraging Therapeutic Proteins and Peptides from Lumbricus Earthworms: Targeting SOCS2 E3 Ligase for Cardiovascular Therapy through Molecular Dynamics Simulations.

Suppressor of cytokine signaling 2 (SOCS2), an E3 ubiquitin ligase, regulates the JAK/STAT signaling pathway, essential for cytokine signaling and immune responses. Its dysregulation contributes to cardiovascular diseases (CVDs) by promoting abnormal cell growth, inflammation, and resistance to cell death. This study aimed to elucidate the molecular mechanisms underlying the interactions between Lumbricus-derived proteins and peptides and SOCS2, with a focus on identifying potential therapeutic candidates for CVDs. Utilizing a multifaceted approach, advanced computational methodologies, including 3D structure modeling, protein-protein docking, 100 ns molecular dynamics (MD) simulations, and MM/PBSA calculations, were employed to assess the binding affinities and functional implications of Lumbricus-derived proteins on SOCS2 activity. The findings revealed that certain proteins, such as Lumbricin, Chemoattractive glycoprotein ES20, and Lumbrokinase-7T1, exhibited similar activities to standard antagonists in modulating SOCS2 activity. Furthermore, MM/PBSA calculations were employed to assess the binding free energies of these proteins with SOCS2. Specifically, Lumbricin exhibited an average ΔGbinding of -59.25 kcal/mol, Chemoattractive glycoprotein ES20 showed -55.02 kcal/mol, and Lumbrokinase-7T1 displayed -69.28 kcal/mol. These values suggest strong binding affinities between these proteins and SOCS2, reinforcing their potential therapeutic efficacy in cardiovascular diseases. Further in vitro and animal studies are recommended to validate these findings and explore broader applications of Lumbricus-derived proteins.

Evaluating the toxicity effects of thiamethoxam and its main metabolite clothianidin to earthworms (Eisenia fetida) from the perspective of endogenous metabolites.

The widespread application of neonicotinoid insecticides (NNIs) has attracted widespread attention to their potential ecotoxicological effects. In this study, we systematically evaluated the toxic effects of thiamethoxam (TMX) and its metabolite clothianidin (CLO) on earthworms (Eisenia fetida). Specifically, the antioxidant system responses and endogenous metabolite metabolism responses in earthworms were analyzed in the temporal dimension after 7, 14, 21 and 28 days of exposure to TMX and CLO. The results found that TMX and CLO could inhibit the growth phenotype of earthworms and cause significant changes in antioxidant system related indicators. More importantly, we found that TMX and CLO could cause significant changes in the metabolic profiles of earthworms through NMR-based metabolomics. From the changes in endogenous metabolites, the toxicity effects of TMX on earthworms gradually increases with prolonged exposure time. Differently, the toxicity effects of CLO on earthworms is significantly higher than that of TMX in the early stages of exposure. Meanwhile, these impacts will not weaken with prolonged exposure time. Furthermore, the results of KEGG enrichment pathway analysis indicated that TMX and CLO could significantly interfere with energy homeostasis, redox homeostasis, osmotic regulation, amino acid metabolism and protein synthesis in earthworms. These findings further deepen our understanding of the ecotoxicological effects of NNIs on soil organism.

An innovative, sustainable, and environmentally friendly approach for wheat drought tolerance using vermicompost and effective microorganisms: upregulating the antioxidant defense machinery, glyoxalase system, and osmotic regulatory substances.

BACKGROUND: Vermicompost contains humic acids, nutrients, earthworm excretions, beneficial microbes, growth hormones, and enzymes, which help plants to tolerate a variety of abiotic stresses. Effective microorganisms (EM) include a wide range of microorganisms' e.g. photosynthetic bacteria, lactic acid bacteria, yeasts, actinomycetes, and fermenting fungi that can stimulate plant growth and improve soil fertility. To our knowledge, no study has yet investigated the possible role of vermicompost and EM dual application in enhancing plant tolerance to water scarcity. METHODS: Consequently, the current study investigated the effectiveness of vermicompost and EM in mitigating drought-induced changes in wheat. The experiment followed a completely randomized design with twelve treatments. The treatments included control, as well as individual and combined applications of vermicompost and EM at three different irrigation levels (100%, 70%, and 30% of field capacity). RESULTS: The findings demonstrated that the application of vermicompost and/or EM significantly improved wheat growth and productivity, as well as alleviated drought-induced oxidative damage with decreased the generation of superoxide anion radical and hydrogen peroxide. This was achieved by upregulating the activities of several antioxidant enzymes, including superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, glutathione peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase. Vermicompost and/or EM treatments also enhanced the antioxidant defense system by increasing the content of antioxidant molecules such as ascorbate, glutathione, phenolic compounds, and flavonoids. Additionally, the overproduction of methylglyoxal in water-stressed treated plants was controlled by the enhanced activity of the glyoxalase system enzymes; glyoxalase I and glyoxalase II. The treated plants maintained higher water content related to the higher content of osmotic regulatory substances like soluble sugars, free amino acids, glycinebetaine, and proline. CONCLUSIONS: Collectively, we offer the first report that identifies the underlying mechanism by which the dual application of vermicompost and EM confers drought tolerance in wheat by improving osmolyte accumulation and modulating antioxidant defense and glyoxalase systems.

Earthworm mucus contributes significantly to the accumulation of soil cadmium in tomato seedlings.

Whether earthworm mucus affects Cd transport behavior in soil-plant systems remains uncertain. Consequently, this study thoroughly assessed the impacts of earthworm mucus on plant growth and physiological responses, plant Cd accumulation, translocation, and distribution, as well as soil characteristics and Cd fractionation in a soil-plant (tomato seedling) system. Results demonstrated that the earthworm inoculation considerably enhanced plant Cd uptake and decreased plant Cd translocation, the effects of which were appreciably less significant than those of the earthworm mucus. This suggested that earthworm mucus may play a crucial role in the way earthworms influence plant Cd uptake and translocation. Moreover, the artificial mucus, which contained identical inorganic nitrogen contents to those in earthworm mucus, had no significant effect on plant Cd accumulation or translocation, implying that components other than inorganic nitrogen in the earthworm mucus may have contributed significantly to the overall effects of the mucus. Compared with the control, the earthworm mucus most substantially increased the root Cd content, the Cd accumulation amount of root and whole plant, and root Cd BCF by 93.7 %, 221.3 %, 72.2 %, and 93.7 %, respectively, while notably reducing the Cd TF by 48.2 %, which may be ascribed to the earthworm mucus's significant impacts on tomato seedling growth and physiological indicators, its considerable influences on the subcellular components and chemical species of root Cd, and its substantial effects on the soil characteristics and soil Cd fractionation, as revealed by correlation analysis. Redundancy analysis further suggested that the most prominent impacts of earthworm mucus may have been due to its considerable reduction of soil pH, improvement of soil DOC content, and enhancement of the exchangeable Cd fraction in soil. This work may help better understand how earthworm mucus influences the transport behavior of metals in soil-plant systems.

Freeze-thaw aging increases the toxicity of microplastics to earthworms and enriches pollutant-degrading microbial genera.

Freeze-thaw (FT) aging can change the physicochemical characteristics of microplastics (MPs). The toxic impacts of FT-aged-MPs to soil invertebrates are poorly understood. Here the toxic mechanisms of FT-aged-MPs were investigated in earthworms after 28 d exposure. Results showed that FT 50 µm PE-MPs significantly increased reactive oxygen species (ROS) by 5.78-9.04 % compared to pristine 50 µm PE-MPs (41.80-45.05 ng/mgprot), whereas FT 500 µm PE-MPs reduced ROS by 7.52-7.87 % compared to pristine 500 µm PE-MPs (51.44-54.46 ng/mgprot). FT-PP-MPs significantly increased ROS and malondialdehyde (MDA) content in earthworms by 14.82-44.06 % and 46.75-110.21 %, respectively, compared to pristine PP-MPs (40.56-44.66 ng/mgprot, 0.41-2.53 nmol/mgprot). FT-aged PE- and PP-MPs caused more severe tissue damage to earthworms. FT-aged PE-MPs increased the alpha diversity of the gut flora of earthworms compared to pristine MPs. Earthworm guts exposed to FT-aged-MPs were enriched with differential microbial genera of contaminant degradation capacity. FT-PE-MPs affected membrane translocation by up-regulating lipids and lipid-like molecules, whereas FT-PP-MPs changed xenobiotic biodegradation and metabolism by down-regulating organoheterocyclic compounds compared to the pristine PE- and PP-MPs. This study concludes that FT-aged MPs cause greater toxicity to earthworms compared to pristine MPs.

Multifunctional biomolecular corona-inspired nanoremediation of antibiotic residues.

Facing complex and variable emerging antibiotic pollutants, the traditional development of functional materials is a "trial-and-error" process based on physicochemical principles, where laborious steps and long timescales make it difficult to accelerate technical breakthroughs. Notably, natural biomolecular coronas derived from highly tolerant organisms under significant contamination scenarios can be used in conjunction with nanotechnology to tackling emerging contaminants of concern. Here, super worms (Tubifex tubifex) with high pollutant tolerance were integrated with nano-zero valent iron (nZVI) to effectively reduce the content of 17 antibiotics in wastewater within 7 d. Inspired by the synergistic remediation, nZVI-augmented worms were constructed as biological nanocomposites. Neither nZVI (0.3 to 3 g/L) nor worms (104 to 105 per liter) alone efficiently degraded florfenicol (FF, as a representative antibiotic), while their composite removed 87% of FF (3 µmol/L). Under antibiotic exposure, biomolecules secreted by worms formed a corona on and modified the nZVI particle surface, enabling the nano-bio interface greater functionality, including responsiveness, enrichment, and reduction. Mechanistically, FF exposure activated glucose-alanine cycle pathways that synthesize organic acids and amines as major metabolites, which were assembled into vesicles and secreted, thereby interacting with nZVI in a biologically response design strategy. Lactic acid and urea formed hydrogen bonds with FF, enriched analyte presence at the heterogeneous interface. Succinic and lactic acids corroded the nZVI passivation layer and promoted electron transfer through surface conjugation. This unique strategy highlights biomolecular coronas as a complex resource to augment nano-enabled technologies and will provide shortcuts for rational manipulation of nanomaterial surfaces with coordinated multifunctionalities.

Roxarsone reduces earthworm-mediated nutrient cycling by suppressing aggregate formation and enzymic activity in soil with manure application.

The application of manure and earthworms are frequently used in fertilization practices to improve C, N, and P cycling in soil, which may be adversely affected by roxarsone (ROX), as an organoarsenical pollutant. To effectively address this issue, in this work, the interactive impacts of ROX and earthworm Eisenia foetida on the aggregate formation, input of organic carbon (OC), and changes in the available N and P following 56-day cultivation were systematically investigated. Compared to the control, earthworms increased the mean weight diameter (MWD) of the soil aggregates from 0.6 to 1.1 mm. Thereby, they activated soil enzymes including catalase (CAT), sucrase (SC), urease (UE), and neutral phosphatase (NP), with the soil's pH decreased to 7.1. Consequently, the values of OC, soluble nitrite (NO3-N), and Olsen-P content were respectively increased by 0.78-, 1.69-, and 0.87- folds in the E treatment (14.3 vs. 25.5 g/kg, 12.8 vs. 33.3 mg/kg, and 7.8 vs. 14.6 mg/kg). Although the changes in the R treatment were slight, ROX reduced the earthworm-mediated improvements of soil fertility during the application of the RE treatment compared to the E treatment, i.e., the values of MWD, OC, NO3-N, and Olsen-P were reduced to 0.9 mm, 20.4 g/kg, 25.4 mg/kg, and 11.6 mg/kg, respectively. From the well-fitted structural equation models, it was demonstrated that earthworms enhanced the aggregate formation and nutrient cycling of OC, NO3-N, and Olsen-P, which were inhibited by ROX. Overall, these adverse effects can be offset by earthworm addition, which can play the dual role of monitor and driver for the soil properties. Our work provides insightful strategies for ROX-bearing manure management.

Differential inducibility of transmembrane peptidoglycan recognition proteins (PGRPs) by bacterial challenges in the earthworm, Eisenia andrei.

Peptidoglycan recognition proteins (PGRPs) and Toll-like receptors (TLRs) are highly conserved pattern recognition receptors (PRRs). Earthworms possess genes encoding TLRs that specifically respond to Gram-positive bacteria. In addition, several PGRPs have been recently identified, which are predicted to exhibit amidase activity but lack receptor function. In lophotrochozoans, a membrane-bound PRR responsible for detecting Gram-negative bacteria remains unidentified. This study reveals several novel transmembrane peptidoglycan recognition proteins (Ean-PGRPLs) in earthworms, whose mRNA expression increases in response to Gram-negative but not Gram-positive bacteria. This indicates that Ean-PGRPLs may serve as a PRR associated with intracellular signaling for Gram-negative bacteria.

Novel Insights into Stereoselective Reproductive Toxicity Induced by Mefentrifluconazole in Earthworms (Eisenia fetida): First Report of Estrogenic Effects.

Widespread use of the new chiral triazole fungicide mefentrifluconazole (MFZ) poses a threat to soil organisms. Although triazole fungicides have been reported to induce reproductive disorders in vertebrates, significant research gaps remain regarding their impact on the reproductive health of soil invertebrates. Here, reproduction-related toxicity end points were explored in earthworms (Eisenia fetida) after exposure for 28 d to soil containing 4 mg/kg racemic MFZ, R-(-)-MFZ, and S-(+)-MFZ. The S-(+)-MFZ treatment resulted in a more pronounced reduction in the number of cocoons and juveniles compared to R-(-)-MFZ treatment, and the expression of annetocin gene was significantly downregulated following exposure to both enantiomers. This reproductive toxicity has been attributed to the disruption of ovarian steroidogenesis at the transcriptional level. Further studies revealed that MFZ enantiomers were able to activate the estrogen receptor (ER). Indirect evidence for this estrogenic effect is provided by the introduction of 17ß-estradiol, which also induces reproductive disorders through ER activation.

Development of a Simple Cost Effective Oxygenation System for In Vivo Solution State NMR in 10 mm NMR Tubes.

In vivo NMR is evolving into an important tool to understand biological processes and environmental responses. Current approaches use flow systems to sustain the organisms with oxygenated water and food (e.g., algae) inside the NMR. However, such systems have the potential to leak and clog (potentially damaging costly hardware), require large volumes of media, and multiple expensive HPLC pumps. The proposed "oxygenation system", uses a simple "double slit" adapter and a single air/oxygen flow line into the NMR. The design is especially suited to larger diameter probes given that standard flow systems would require higher flow rates thus amplifying the potential and impact of leaks/clogs. Traditionally, in vivo NMR of small organisms (e.g., Daphnia) have required 2D NMR in combination with 13C enrichment to overcome susceptibility distortions and provide information rich metabolic profiles. Here Daphnia magna, Eisenia fetida and Artemia franciscana are used to demonstrate the potential of the oxygenation system. Survivability tests and 1H time-resolved monitoring were first performed on D. magna, while E. fetida contained enough biomass to permit 1H-13C HSQC, 13C-1H HETCOR and 31P NMR without isotopic enrichment. Finally, STOCSY of 1D 13C NMR was used to follow the growth of A. franciscana (without 13C enrichment) for 48 h after birth, which helps visualize trends across a series of 1D in vivo data. In summary, application of the oxygenation system toward larger diameter probes allows the collection of NMR data without enrichment, offering a promising solution to better understand processes in vivo.

Accumulation of heavy metals from single and combined olive mill wastewater and pomace in soil and bioaccumulation in tissues of two earthworm species: Endogeic (Aporrectodea trapezoides) and Epigeic (Eisenia fetida).

Soil and earthworms are threatened by anthropogenic contamination resulting from olive mill waste dumping on the soil due to their pollutant properties. While several studies have explored the effects of olive mill waste on soil properties and the accumulation of heavy metals in soil, there is currently a gap in the literature regarding the potential bioaccumulation of heavy metals from olive mill waste in earthworms. In this study, soil with earthworms from two ecological categories (endogeic: Aporrectodea trapezoides and epigeic: Eisenia fetida) was treated with increasing doses of olive mill wastewater (OMWW) and olive mill pomace (OMP), applied individually or combined, in an indoor experiment in plastic containers, under laboratory conditions. The results revealed the presence of significant concentrations of heavy metals in the two types of wastes ranging as follows: Fe˃ Zn˃ Cu˃ Cd˃ Cr for OMWW, and Fe˃ Zn˃ Cu˃ Cr for OMP (with Cd below the detection limit). The study demonstrated distinct effects of OMWW and OMP, both individually and in combination, on soil heavy metal content, ranging as follows: soil OMWW > soil Combination > soil OMP for Cd; soil Combination > soil OMWW > soil OMP for Cr and Fe; and soil Combination > soil OMP > soil OMWW for Cu and Zn. Additionally, our investigation showed that both earthworm species exhibited significant uptake of these metals into their tissues, particularly the endogeic species. Interestingly, the most significant difference between species was in the accumulation of Cu, with the epigeic species accumulating significantly lower amounts.

Differential eco-toxicological responses toward Eisenia fetida exposed to soil contaminated with naphthalene and typical metabolites.

Naphthalene (NAP) was frequently detected in polycyclic aromatic hydrocarbons (PAHs)-contaminated soil, and its residues may pose an eco-toxicological threat to soil organisms. The toxic effects of NAP were closely tied to phenolic and quinone metabolites in biological metabolism. However, the present knowledge concerning the eco-toxicological impacts of NAP metabolites at the animal level is scanty. Here, we assessed the differences in the eco-toxicological responses of Eisenia fetida (E. fetida) in NAP, 1-naphthol (1-NAO) or 1,4-naphthoquinone (1,4-NQ) contaminated soils. NAP, 1-NAO, and 1,4-NQ exposure triggered the onset of oxidative stress as evidenced by the destruction of the antioxidant enzyme system. The lipid peroxidation and DNA oxidative damage levels induced by 1-NAO and 1,4-NQ were higher than those of NAP. The elevation of DNA damage varied considerably depending on differences in oxidative stress and the direct mode of action of NAP or its metabolites with DNA. All three toxicants induced different degrees of physiological damage to the body wall, but only 1, 4-NQ caused the shedding of intestinal epithelial cells. The integrated biomarker response for different exposure times illustrated that the comprehensive toxicity at the animal level was 1,4-NQ > 1-NAO > NAP, and the time-dependent trends of oxidative stress responses induced by the three toxicants were similar. At the initial stage, the antioxidant system of E. fetida responded positively to the provocation, but the ability of E. fetida to resist stimulation decreased with the prolongation of time resulting in provocation oxidative damage. This study would provide new insights into the toxicological effects and biohazard of PAHs on soil animals.

Toxicological analysis of Eisenia fetida in soil under the coexistence of rockwool substrate andtricyclazole.

This study investigated the combined effects of rockwool, a novel seedling substrate, and tricyclazole (TCA) on the bioavailability of TCA to Eisenia fetida. The single addition of rockwool and TCA alone to the soil inhibited the growth of E. fetida. A high concentration (300 mg·L-1) of TCA significantly decreased the biomass of E. fetida. The addition of 20-mesh rockwool reduced this effect on earthworm biomass by decreasing the soil TCA through adsorption, effectively mitigating TCA bioaccumulation in earthworms. A mechanistic analysis showed that the Mg-O functional group on the rockwool surface combined with the CC functional group in TCA to generate Mg-O-C, and the adsorption process was dominated by chemisorption. Toxicology experiments demonstrated that malondialdehyde and cellulase could be used as biomarkers of inhibitory effects of combined rockwool and TCA in soil on E. fetida. Macrogenomic analyses revealed that small particle sizes and high concentrations of rockwool caused co-stress effects on earthworms when TCA was present. When the particle size of rockwool increased, the toxic effect of TCA on earthworms instead decreased at higher rockwool concentrations. Therefore, in practical agricultural production, the particle size of rockwool can be controlled to realize the adsorption of TCA and reduce the toxic effects of TCA and rockwool on earthworms.

Earthworm injury test for in-situ biomonitoring of pesticides in biobeds.

Biobeds are presented as an alternative for good pesticide wastewater management on farms. This work proposes a new test for in-situ biomonitoring of pesticide detoxification in biobeds. It is based on the assessment of visually appreciable injuries to Eisenia fetida. The severity of the injury to each exposed individual is assessed from the morphological changes observed in comparison with the patterns established in seven categories and, an injury index is calculated. A linear relationship between the proposed injury index and the pesticide concentration was determined for each pesticide sprayed individually in the biomixture. The five pesticides used were atrazine, prometryn, clethodim, haloxyfop-P-methyl and dicamba. In addition, a multiple linear regression model (i.e., a multivariate response surface) was fitted, which showed a good generalization capacity. The sensitivity range of the injury test was tested from 0.01 to 630 mg kg-1 as the total pesticide concentration. This index is then used to monitor the detoxification of these pesticides in a biomixture (composed of wheat stubble, river waste, and soil, 50:25:25% by volume) over 210 days. The results are compared with standardized tests (Eisenia fetida avoidance test and Lactuca sativa seed germination test) carried out on the same biomixture. The results are also compared with data on the removal of pesticides. The injury test showed a better correlation with the removal of pesticides than the avoidance test and seed germination test. This simple and inexpensive test has proved to be useful for decontamination in-situ monitoring in biobeds.

Co-exposure to boscalid and amoxicillin inhibited the degradation of boscalid and aggravated the threat to the earthworm.

The extensive application of pesticides and antibiotics in agricultural production makes it possible for them to coexist in farmland, and the interaction of the two pollutants can lead to changes in environmental behavior and toxicity, creating uncertainty risks to soil and soil organisms. In this study, we explored the environmental behavior and the effects of earthworms under co-exposure to amoxicillin and boscalid and further explored the accumulation and toxic effects on earthworms. The results showed that amoxicillin increased the adsorption of boscalid in soil and inhibited its degradation. In addition, we noticed that the co-exposure of amoxicillin and boscalid caused intestinal barrier damage, which increased the bioaccumulation of earthworms for boscalid and led to more severe oxidative stress and metabolic disorders in earthworms. In summary, our findings indicate that amoxicillin can increase the ecological risk of boscalid in the environment and imply that the encounter between antibiotics and pesticides in the environment can amplify the toxic effects of pesticides, which provides new insights into the ecological risks of antibiotics.

Silver sulfide nanoparticles eliminate the stimulative effects of earthworms on nutrient uptake by soybeans in high organic matter soils.

A significant knowledge gap exists regarding the impact of soil organic matter on the bioavailability of Ag2S-NPs (environmentally relevant forms of Ag-NPs) in soil-earthworm-plant systems. This study used two soils with varying organic matter content, both with and without earthworms, to investigate the bioavailability of Ag2S-NPs. The findings revealed an 80 % increase in Ag bioaccessibility to soybeans in soils with high organic matter content compared to soils with low organic matter. Additionally, the presence of earthworms significantly increased Cl concentrations from 24.3-62.2 mg L-1 to 80.1-147.2 mg L-1, triggering the elevated bioavailability of Ag. Interestingly, Ag2S-NPs eliminated the stimulative effects of earthworms on plant nutrient uptake. In the presence of earthworms, the high organic matter soil amended with Ag2S-NPs exhibited lower concentrations of essential elements (Ca, Cu, Fe, K, and P) in plant tissues compared to soils without earthworms. Our study presents evidence of the transformation of Ag2S-NPs into Ag-NPs across various soil solutions, resulting in the formation of Ag nanoparticle complexes. Particularly noteworthy is the significant reduction in particle sizes in soils incubated with earthworms and high organic matter content, from 85.0 nm to 40.2 nm. Notably, in the rhizosphere soil, a decrease in the relative abundance of nutrient cycling-related phyla was observed, with reductions of 18.5 % for Proteobacteria and 30.0 % for Actinobacteriota. These findings offer valuable insights into the biological and biochemical consequences of Ag2S-NP exposure on earthworm-mediated plant nutrient acquisition.

Soil amendments reduce PFAS bioaccumulation in Eisenia fetida following exposure to AFFF-impacted soil.

The efficacy of RemBind® 300 to immobilize per- and polyfluoroalkyl substances (PFAS) in aqueous film forming foam (AFFF)-impacted soil (∑28 PFAS 1280-8130 ng g-1; n = 8) was assessed using leachability (ASLP) and bioaccumulation (Eisenia fetida) endpoints as the measure of efficacy. In unamended soil, ∑28 PFAS leachability ranged from 26.0 to 235 µg l-1, however, following the addition of 5% w/w RemBind® 300, ∑28 PFAS leachability was reduced by > 99%. Following exposure of E. fetida to unamended soil, ∑28 PFAS bioaccumulation ranged from 18,660-241,910 ng g-1 DW with PFOS accumulating to the greatest extent (15,150-212,120 ng g-1 DW). Biota soil accumulation factors (BSAF) were significantly (p < 0.05) higher for perfluoroalkyl sulfonic acids (PFSA; 13.2-50.9) compared to perfluoroalkyl carboxylic acids (PFCA; 1.2-12.7) while for individual PFSA, mean BSAF increased for C4 to C6 compounds (PFBS: 42.6; PFPeS: 52.7; PFHxS: 62.4). In contrast, when E. fetida were exposed to soil amended with 5% w/w RemBind® 300, significantly lower PFAS bioaccumulation occurred (∑28 PFAS: 339-3397 ng g-1 DW) with PFOS accumulation 23-246 fold lower compared to unamended soil. These results highlight the potential of soil amendments for reducing PFAS mobility and bioavailability, offering an immobilization-based risk management approach for AFFF-impacted soil.

Environmentally relevant concentrations of microplastics from agricultural mulch and cadmium negatively impact earthworms by triggering neurotoxicity and disrupting homeostasis.

Recent research has highlighted the ecological risk posed by microplastics (MPs) from mulching film and heavy metals to soil organisms. However, most studies overlooked real environmental levels of MPs and heavy metals. To address this gap, pristine and aged polyethylene (PE) mulching film-derived MPs (PMPs, 500 mg/kg; AMPs, 500 mg/kg) were combined with cadmium (Cd, 0.5 mg/kg) to assess the acute toxicity to earthworms and investigate associated molecular mechanisms (oxidative stress, osmoregulation pressure, gut microbiota, and metabolic responses) at environmentally relevant concentrations. Compared to Cd alone and Cd + PMPs treatments (11.15 ± 4.19 items/g), Cd + AMPs treatment resulted in higher MPs bioaccumulation (23.73 ± 13.14 items/g), more severe tissue lesions, and increased cell membrane osmotic pressure in earthworms' intestines. Cd + AMPs induced neurotoxicity through elevated levels of glutamate and acetylcholinesterase. Earthworm intestines (0.98 ± 0.49 to 3.33 ± 0.37 mg/kg) exhibited significantly higher Cd content than soils (0.19 ± 0.01 to 0.51 ± 0.06 mg/kg) and casts (0.15 ± 0.01 to 0.25 ± 0.05 mg/kg), indicating PE-MPs facilitated Cd transport in earthworms' bodies. Metabolomic analysis showed Cd + AMPs exposure depleted energy and nucleotide metabolites, disrupted cell homeostasis more profoundly than Cd and Cd + PMPs treatments. Overall, co-exposure to AMPs + Cd induced more severe neurotoxicity and disruption of homeostasis in earthworm than Cd and PMPs + Cd treatments. Our study, using Cd and MPs with environmental relevance, underscores MPs' role in amplifying Cd accumulation and toxicity in earthworms.