Impact of pesticides on non-target invertebrates in agricultural ecosystems.

In fact, less than 1% of applied pesticides reach their target pests, while the remainder pollute the neighboring environment and adversely impact human health as well as non-target organisms in agricultural ecosystem. Pesticides can contribute to the loss of agrobiodiversity, which are essential to maintaining the agro-ecosystem's structure and functioning in order to produce and secure enough food. This review article examines the negative effects of pesticides on non-target invertebrates including earthworms, honeybees, predators, and parasitoids. It also highlights areas where further research is needed to address unresolved issues related to pesticide exposure, aiming to improve conservation efforts for these crucial species. These organisms play crucial roles in ecosystem functioning, such as soil health, pollination, and pest control. Both lethal and sub-lethal effects of pesticides on the selected non-target invertebrates were discussed. Pesticides affect DNA integrity, enzyme activity, growth, behavior, and reproduction of earthworms even at low concentrations. Pesticides could also induce a reduction in individual survival, disruption in learning performance and memory, as well as a change in the foraging behavior of honeybees. Additionally, pesticides adversely affect population growth indices, reproduction, development, longevity, and consumption of predators and parasitoids. As a result, pesticides must pass adequate ecotoxicological risk assessment to be enlisted by regulatory authorities. Therefore, it is important to adopt integrated pest management (IPM) strategies that minimize pesticide use and promote the conservation of beneficial organisms in order to maintain agrobiodiversity and sustainable agricultural systems. Furthermore, adopting precision agriculture and organic farming lessen these negative effects as well.less than.

Nickel oxide nanoparticles decrease the accumulation of atrazine in earthworms.

Nickel oxide nanoparticles (NiO-NPs) are common nanomaterials that may be released into the environment, affecting the toxicity of other contaminants. Atrazine (ATZ) is a commonly used herbicide that can harm organisms due to its persistence and bioaccumulation in the environment. Although the toxicity of ATZ to earthworms is well-documented, the risk of co-exposure with NiO-NPs increases as more nanoparticles accumulate in the soil. In this study, we investigated the effects and mechanisms of NiO-NPs on the accumulation of ATZ in earthworms. The results showed that after day 21, the antioxidant system of the cells under ATZ treatment alone was adversely affected, with ROS content 36.05 % higher than that of the control (CK) group. However, the addition of NiO-NPs reduced the ROS contents in the earthworms by 0.6 %- 32.3 %. Moreover, analysis of earthworm intestinal sections indicates that NiO-NPs mitigated cellular and tissue damage caused by ATZ. High-throughput sequencing revealed that NiO-NPs in earthworm intestines increased the abundance of Pseudomonas aeruginosa and Aeromonas aeruginosa. Additionally, the enhanced function of the ABC transport system in the gut resulted in lower accumulation of ATZ in earthworms. In summary, NiO-NPs can reduce the accumulation and thus the toxicity of ATZ in earthworms. Our study contributes to a deeper understanding of the effects of NiO-NPs on co-existing pollutants.

Novel mechanistic insights into Cr(VI) and Cr(III) induced discrepancies of cellular toxicity and oxidative injury events in Eisenia fetida.

Chromium (Cr) poses a high ecological risk, however the toxic mechanisms of Cr in different valence states to soil organisms at cellular and molecular levels are not exactly. In this study, the Eisenia fetida coelomocytes and Cu/Zn-superoxide dismutase (Cu/Zn-SOD) were chosen as the target subjects to investigate the effects and mechanisms of cellular toxicity induced by Cr(VI) and Cr(III). Results indicated that Cr(VI) and Cr(III) significantly reduced the coelomocytes viability. The level of reactive oxygen species (ROS) was markedly increased after Cr(VI) exposure, which finally reduced antioxidant defense abilities, and induced lipid peroxidation and cellular membrane damage in earthworm coelomocytes. However, Cr(III) induced lower levels of oxidative stress and cellular damage with respect to Cr(VI). From a molecular perspective, the binding of both Cr(VI) and Cr(III) with Cu/Zn-SOD resulted in protein backbone loosening and reduced ß-Sheet content. The Cu/Zn-SOD showed fluorescence enhancement with Cr(III), whereas Cr(VI) had no obvious effect. The activity of Cu/Zn-SOD continued to decrease with the exposure of Cr. Molecular docking indicated that Cr(III) interacted more readily with the active center of Cu/Zn-SOD. Our results illustrate that oxidative stress induced by Cr(VI) and Cr(III) plays an important role in the cytotoxic differences of Eisenia fetida coelomocytes and the binding of Cr with Cu/Zn-SOD can also affect the normal structures and functions of antioxidant defense-associated protein.

Multilevel investigation of the ecotoxicological effects of sewage sludge biochar on the earthworm Eisenia fetida.

The ecological risks of sewage sludge biochar (SSB) after land use is still not truly reflected. Herein, the ecological risks of SSB prepared at different temperature were investigated using the earthworm E. fetida as a model organism from the perspectives of organismal, tissue, cellular, and molecular level. The findings revealed that the ecological risk associated with low-temperature SSB (SSB300) was more pronounced compared to medium- and high-temperature SSB (SSB500 and SSB700), and the ecological risk intensified with increasing SSB addition rates, as revealed by an increase in the integrated biomarker response v2 (IBRv2) value by 2.59-25.41 compared to those of SSB500 and SSB700. Among them, 10% SSB300 application caused significant oxidative stress and neurotoxicity in earthworms compared to CK (p < 0.001). The weight growth rate and cocoon production rate of earthworms were observed to decrease by 25.06% and 69.29%, respectively, while the mortality rate exhibited a significant increase of 33.34% following a 10% SSB300 application, as compared to the CK. Moreover, 10% SSB300 application also resulted in extensive stratum corneum injury and significant longitudinal muscle damage in earthworms, while also inducing severe collapse of intestinal epithelial cells and disruption of intestinal integrity. In addition, 10% SSB300 caused abnormal expression of earthworm detoxification and cocoon production genes (p < 0.001). These results may improve our understanding of the ecotoxicity of biochar, especially in the long term application, and contribute to providing the guidelines for applying biochar as a soil amendment.

Co-exposure of decabromodiphenyl ethane and cadmium increases toxicity to earthworms: Enrichment, oxidative stress, damage and molecular binding mechanisms.

The increase of electronic waste worldwide has resulted in the exacerbation of combined decabromodiphenyl ethane (DBDPE) and cadmium (Cd) pollution in soil, posing a serious threat to the safety of soil organisms. However, whether combined exposure increases toxicity remains unclear. Therefore, this study primarily investigated the toxic effects of DBDPE and Cd on earthworms at the individual, tissue, and cellular levels under single and combined exposure. The results showed that the combined exposure significantly increased the enrichment of Cd in earthworms by 50.32-90.42 %. The toxicity to earthworms increased with co-exposure, primarily resulting in enhanced oxidative stress, inhibition of growth and reproduction, intensified intestinal and epidermal damage, and amplified coelomocyte apoptosis. PLS-PM analysis revealed a significant and direct relationship between the accumulation of target pollutants in earthworms and oxidative stress, damage, as well as growth and reproduction of earthworms. Furthermore, IBR analysis indicated that SOD and POD were sensitive biomarkers in earthworms. Molecular docking elucidated that DBDPE and Cd induced oxidative stress responses in earthworms through the alteration of the conformation of the two enzymes. This study enhances understanding of the mechanisms behind the toxicity of combined pollution and provides important insights for assessing e-waste contaminated soils.

Elevated temperature magnifies the acute and chronic toxicity of clothianidin to Eisenia fetida.

Pesticide residue and thermal stress resulting from global climate change are parallel stressors for soil fauna. However, it remains ambiguous how elevated temperatures and pesticides can interact to threaten soil fauna. In the study, the acute and chronic clothianidin (CTD) toxicity to earthworms (Eisenia fetida) at different temperatures, and the effect of increasing temperature on antioxidant defense mechanisms in response to CTD were investigated. The acute toxicity of CTD was exacerbated by increased temperature in both filter paper contact tests (a decrease in the 48-h median lethal concentration (LC50) from 0.077 µg/cm2 at 20 °C to 0.009 µg/cm2 at 30 °C) and natural soil tests (a decrease in the 48-h LC50 from 0.774 mg/kg at 20 °C to 0.199 mg/kg at 30 °C). Exposure to CTD or high temperature (30 °C) triggered reactive oxygen species (ROS) overgeneration and increased antioxidant enzyme activities in earthworms; and the effect was particularly pronounced after exposure to both higher temperatures and CTD. At 20 and 25 °C, there was no significant change in the growth and reproduction of E. fetida after 56-d exposure to CTD-contaminated soil. However, the combined effect of CTD and high temperature (30 °C) significantly reduced the weight change rate, cocoon number, hatching rate, and number of juveniles on day 56. These results indicated that elevated temperature could aggravate acute and chronic CTD toxicity to earthworms. The findings emphasize that evaluating changes in pesticide toxicity under global warming is worth further investigation.

Bioconversion of cow manure through vermicomposting: effects of tylosin concentration on the weight of worms and manure quality.

This study investigated batch-fed vermicomposting of cow manure, with a specific focus on assessing the effects of tylosin on the weight of earthworms and the overall quality of the resulting manure. Five reactors, including three concentrations of tylosin (50, 100, and 150 mg/kg) and two control reactors, were employed. Residual tylosin concentrations were measured using high-performance liquid chromatography (HPLC). Quality parameters such as pH, temperature, volatile solids (VS), organic carbon content (OCC), electrical conductivity (EC), ash content, C/N ratio, total Kjeldahl nitrogen (TKN), and microbial content were evaluated. The toxicity and maturity of vermicompost were assessed by determining the germination index (GI). The study also monitored variations in the earthworm's weight. The results demonstrated a decreasing trend in VS, OCC, C/N, and fecal coliforms, along with increased pH, EC, ash content, and TKN during the vermicomposting process. Furthermore, investigations revealed significant reductions in the reactors with tylosin concentrations of 50, 100, and 150 mg/kg, resulting in the removal of 98%, 90.48%, and 89.38% of the initial tylosin, respectively. This result confirms the faster removal of tylosin in reactors with lower concentrations. Degradation of tylosin also conforms to first-order kinetics. The findings showed a significant influence of tylosin on the weight of Eisenia fetida earthworms and the lowest antibiotic concentration led to the highest weight gain. Finally, the high percentage of germination index (90-100%) showed that the quality and maturity of vermicompost is by national and international standards.

Combined effects of cadmium and sulfamethoxazole on Eisenia fetida: Insights into accumulation, subcellular partitioning, biomarkers and toxicological responses.

Cadmium (Cd) and sulfamethoxazole (SMX) frequently coexist in farmlands, yet their synergistic toxicological impacts on terrestrial invertebrates remain unexplored. In this study, earthworms were exposed to artificial soils percolated with Cd (5 mg/kg), SMX (5 mg/kg) or combination of them for 7 days, followed by a 12-day elimination phase in uncontaminated soil. The uptake of Cd and SMX by the earthworms, along with their subcellular distribution, was meticulously analyzed. Additionally, a suite of biomarkers-including superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and weight loss-were evaluated to assess the health status of the earthworms and the toxicological effects of the Cd and SMX mixture. Notably, the cotreatment with Cd and SMX resulted in a significantly higher weight loss in Eisenia fetida (41.25 %) compared to exposure to Cd alone (26.84 %). Moreover, the cotreatment group exhibited substantially higher concentrations of Cd in the total internal body, fraction C (cytosol), and fraction E (tissue fragments and cell membranes) in Eisenia fetida compared to Cd alone counterparts. The combined exposure also significantly elevated the SMX levels in the total body and fraction C compared with the SMX-only treated earthworms. Additionally, Eisenia fetida subjected to the combined treatment showed markedly increased activities of SOD, CAT, and MDA compared to those treated with Cd alone. The effect addition indices (EAIs), ranging from 1.00 to 2.23, unequivocally demonstrated a synergistic effect of the combined treatments. Interestingly, relocating the earthworms to clean soil did not mitigate the observed adverse effects. These findings underscore the increased risk posed by the Cd-SMX complex to terrestrial invertebrates in agricultural areas.

Joint toxic mechanism of clothianidin and prochloraz in the earthworm (Eisenia fetida).

Pesticides are typically present as combinations within soil ecosystems and have detrimental effects on untamed surroundings. However, the collective impacts and fundamental mechanisms of pesticides on soil living beings are currently inadequately assessed. In our current work, we evaluated the interactive consequences of clothianidin (CLO) and prochloraz (PRO) on earthworms (Eisenia fetida) using several toxicological tests, such as acute adverse effects, biocatalytic activity, and alterations in transcriptional activity. The findings revealed that CLO (with a 14-day LC50 value of 6.08 mg kg-1) exhibited greater toxicity compared to PRO (with a 14-day LC50 value of 79.41 mg kg-1). Moreover, the combinations of CLO and PRO had synergistic acute effects on E. fetida. Additionally, the activities of POD, CAT, and GST were significantly varied in most instances of single and mixed treatments when compared to the control. Surprisingly, the transcriptional levels of four genes (gst, sod, crt, and ann), related to oxidative load, metabolic detoxification systems, endoplasmic reticulum, and oxytocin neuropeptide, respectively, were also altered in response to single and mixture exposures, as compared to the control. Alterations in enzyme activity and gene transcriptional level could serve as early indicators for detecting co-exposure to pesticides. The findings of this research offered valuable holistic understanding regarding the toxicity of pesticide combinations on earthworms. Further research should be conducted to investigate the persistent effects of pesticide mixtures on terrestrial invertebrates in order to draw definitive conclusions about the associated risks.

Effect of conventional and biodegradable microplastics on earthworms during vermicomposting process.

The potential effect of microplastics is an increasingly growing environmental issue. However, very little is known regarding the impact of microplastics on the vermicomposting process. The present study explored the effect of non-biodegradable (low density polyethylene; LDPE) and biodegradable (polybutylene succinate-co-adipate; PBSA) microplastics on earthworm Eisenia fetida during vermicomposting of cow dung. For this, earthworms were exposed to different concentrations (0, 0.5, 1 and 2%) of LDPE and PBSA of 2 mm size. The cow dung supported the growth and hatchlings of earthworms, and the toxicity effect of both LDPE and PBSA microplastics on Eisenia fetida was analyzed. Microplastics decreased the body weight of earthworms and there was no impact on hatchlings. The body weight of earthworm decreased from 0 to 60th day by 18.18% in 0.5% of LDPE treatment, 5.42% in 1% of LDPE, 20.58% in 2% of LDPE, 19.99% in 0.5% of PBSA, 15.09% in 1% of PBSA and 16.36% in 2% of PBSA. The physico-chemical parameters [pH (8.55-8.66), electrical conductivity (0.93-1.02 (S/m), organic matter (77.6-75.8%), total nitrogen (3.95-4.25 mg/kg) and total phosphorus (1.16-1.22 mg/kg)] do not show much significant changes with varying microplastics concentrations. Results of SEM and FTIR-ATR analysis observed the surface damage of earthworms, morphological and biochemical changes at higher concentrations of both LDPE and PBSA. The findings of the present study contribute to a better understanding of microplastics in vermicomposting system.

Oxidative stress and DNA alteration on the earthworm Eisenia fetida exposed to four commercial pesticides.

Modern agriculture is mainly based on the use of pesticides to protect crops but their efficiency is very low, in fact, most of them reach water or soil ecosystems causing pollution and health hazards to non-target organisms. Fungicide triazoles and strobilurins based are the most widely used and require a specific effort to investigate toxicological effects on non-target species. This study evaluates the toxic effects of four commercial fungicides Prosaro® (tebuconazole and prothioconazole), Amistar®Xtra (azoxystrobin and cyproconazole), Mirador® (azoxystrobin) and Icarus® (Tebuconazole) on Eisenia fetida using several biomarkers: lipid peroxidation (LPO), catalase activity (CAT), glutathione S-transferase (GST), total glutathione (GSHt), DNA fragmentation (comet assay) and lysozyme activity tested for the first time in E. fetida. The exposure to Mirador® and AmistarXtra® caused an imbalance of ROS species, leading to the inhibition of the immune system. AmistarXtra® and Prosaro®, composed of two active ingredients, induced significant DNA alteration, indicating genotoxic effects. This study broadened our knowledge of the effects of pesticide product formulations on earthworms and showed the need for improvement in the evaluation of toxicological risk deriving from the changing of physicochemical and toxicological properties that occur when a commercial formulation contains more than one active ingredient and several unknown co-formulants.

Toxicity evaluation of neonicotinoids to earthworm (Eisenia fetida) behaviors by a novel locomotion tracking assay.

Pesticides are substances used for controlling, preventing, and repelling pests in agriculture. Among them, neonicotinoids have become the fastest-growing class of insecticides because of their efficiency in targeting pests. They work by strongly binding to nicotinic acetylcholine receptors (nAChRs) in the central nervous system of insects, leading to receptor blockage, paralysis, and death. Despite their selectivity for insects, these substances may be hazardous to non-target creatures, including earthworms. Although earthworms may be invasive in some regions like north America, they contribute to the development of soil structure, water management, nutrient cycling, pollution remediation, and cultural services, positively impacting the environment, particularly in the soil ecosystem. Thus, this study aimed to develop a novel earthworm behavior assay since behavior is a sensitive marker for toxicity assay, and demonstrated its application in evaluating the toxicity of various neonicotinoids. Here, we exposed Eisenia fetida to 1 and 10 ppb of eight neonicotinoids (acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram pestanal, thiacloprid, thiametoxam, and sulfoxaflor) for 3 days to observe their behavior toxicities. Overall, all of the neonicotinoids decreased their locomotion, showed by a reduction of average speed by 24.94-68.63% and increment in freezing time movement ratio by 1.51-4.25 times, and altered their movement orientation and complexity, indicated by the decrement in the fractal dimension value by 24-70%. Moreover, some of the neonicotinoids, which were acetamiprid, dinotefuran, imidacloprid, nitenpyram, and sulfoxaflor, could even alter their exploratory behaviors, which was shown by the increment in the time spent in the center area value by 6.94-12.99 times. Furthermore, based on the PCA and heatmap clustering results, thiametoxam was found as the neonicotinoid that possessed the least pronounced behavior toxicity effects among the tested pesticides since these neonicotinoid-treated groups in both concentrations were grouped in the same major cluster with the control group. Finally, molecular docking was also conducted to examine neonicotinoids' possible binding mechanism to Acetylcholine Binding Protein (AChBP), which is responsible for neurotransmission. The molecular docking result confirmed that each of the neonicotinoids has a relatively high binding energy with AChBP, with the lowest binding energy was possessed by thiametoxam, which consistent with its relatively low behavior toxicities. Thus, these molecular docking results might hint at the possible mechanism behind the observed behavior alterations. To sum up, the present study demonstrated that all of the neonicotinoids altered the earthworm behaviors which might be due to their ability to bind with some specific neurotransmitters and the current findings give insights into the toxicities of neonicotinoids to the environment, especially animals in a soil ecosystem.

Combined toxicity of polyethylene microplastics and nickel oxide nanoparticle on earthworm (Eisenia andrei): oxidative stress responses, bioavailability and joint effect.

The co-occurrence of heavy metals and microplastics (MPs) is an emerging issue that has attracted considerable attention. However, the interaction of nickel oxide nanoparticle (nano-NiO) combined with MPs in soil was poorly researched. Here, experiments were conducted to study the influence of nano-NiO (200 mg/kg) and polyethylene (PE) MPs with different concentrations (0.1, 1, and 10%) and sizes (13, 50, and 500 µm) on earthworms for 28 days. Compared to control, the damage was induced by PE and nano-NiO, which was evaluated by biomarker Integrated Biomarker Response index: version 2 (IBRv2) based on six biomarkers including SOD, POD, CAT, MDA, AChE, Na+/K+-ATPase and cellulase. The majority of the chosen biomarkers showed significant but complicated responses with increasing contaminant concentrations after 28 days of exposure. Moreover, the joint effect was assessed as antagonism by the effect addition index (EAI). Overall, this work expands our understanding of the combined toxicity of PE and nano-NiO in soil ecosystems.

Mixture toxicity study of two metal oxide nanoparticles and chlorpyrifos on Eisenia andrei earthworms.

Co-exposure soil studies of pollutants are necessary for an appropriate ecological risk assessment. Here, we examined the effects of two-component mixtures of metal oxide nanoparticles (ZnO NPs or goethite NPs) with the insecticide chlorpyrifos (CPF) under laboratory conditions in short-term artificial soil assays using Eisenia andrei earthworms. We characterized NPs and their mixtures by scanning electron microscopy, atomic force microscopy, dynamic light scattering and zeta potential, and evaluated effects on metal accumulation, oxidative stress enzymes, and neurotoxicity related biomarkers in single and combined toxicity assays. Exposure to ZnO NPs increased Zn levels compared to control in single and combined exposure (ZnO NPs + CPF) at 72 h and 7 days, respectively. In contrast, there was no indication of Fe increase in organisms exposed to goethite NPs. One of the most notable effects on oxidative stress biomarkers was produced by single exposure to goethite NPs, showing that the worms were more sensitive to goethite NPs than to ZnO NPs. Acetylcholinesterase and carboxylesterase activities indicated that ZnO NPs alone were not neurotoxic to earthworms, but similar degrees of inhibition were observed after single CPF and ZnO NPs + CPF exposure. Differences between single and combined exposure were found for catalase and superoxide dismutase (goethite NPs) and for glutathione S-transferase (ZnO NPs) activities, mostly at 72 h. These findings suggest a necessity to evaluate mixtures of NPs with co-existing contaminants in soil, and that the nature of metal oxide NPs and exposure time are relevant factors to be considered when assessing combined toxicity, as it may have an impact on ecotoxicological risk assessment.

Earthworms improve the rhizosphere micro-environment to mitigate the toxicity of microplastics to tomato (Solanum lycopersicum).

Microplastics (MPs) are widespread in agricultural soil, potentially threatening soil environmental quality and plant growth. However, toxicological research on MPs has mainly been limited to individual components (such as plants, microbes, and animals), without considering their interactions. Here, we examined earthworm-mediated effects on tomato growth and the rhizosphere micro-environment under MPs contamination. Earthworms (Eisenia fetida) mitigated the growth-inhibiting effect of MPs on tomato plant. Particularly, when exposed to environmentally relevant concentrations (ERC, 0.02% w/w) of MPs, the addition of earthworms significantly (p < 0.05) increased shoot and root dry weight by 12-13% and 13-14%, respectively. MPs significantly reduced (p < 0.05) soil ammonium (NH4+-N) (0.55-0.69 mg/kg), nitrate nitrogen (NO3--N) (7.02-8.65 mg/kg) contents, and N cycle related enzyme activities (33.47-42.39 µg/h/g) by 37.7-50.9%, 22.6-37.2%, and 34.2-48.0%, respectively, while earthworms significantly enhanced (p < 0.05) inorganic N mineralization and bioavailability. Furthermore, earthworms increased bacterial network complexity, thereby enhancing the robustness of the bacterial system to resist soil MPs stress. Meanwhile, partial least squares modelling showed that earthworms significantly influenced (p < 0.01) soil nutrients, which in turn significantly affected (p < 0.01) plant growth. Therefore, the comprehensive consideration of soil ecological composition is important for assessing MPs ecological risk.

Toxicity assessment of animal manure composts containing environmental microplastics by using earthworms Eisenia andrei.

Nowadays, animal manure composting constitutes a sustainable alternative for farmers to enhance the level of nutrients within soils and achieve a good productivity. However, pollutants may be present in manures. This study focuses on the detection of environmental microplastics (EMPs) into composts, as well as on the assessment of their potential toxicity on the earthworm Eisenia andrei. To these aims, animals were exposed to two types of compost, namely bovine (cow) and ovine (sheep) manure, besides to their mixture, for 7 and 14 days. The presence and characterization of EMPs was evaluated in all the tested composts, as well as in tissues of the exposed earthworms. The impact of the tested composts was assessed by a multi-biomarker approach including cytotoxic (lysosomal membrane stability, LMS), genotoxic (micronuclei frequency, MNi), biochemical (activity of catalase, CAT, and glutathione-S-transferase, GST; content of malondialdehyde, MDA), and neurotoxic (activity of acetylcholinesterase, AChE) responses in earthworms. Results indicated the presence of high levels of EMPs in all the tested composts, especially in the sheep manure (2273.14 ± 200.89 items/kg) in comparison to the cow manure (1628.82 ± 175.23 items/kg), with the size <1.22 µm as the most abundant EMPs. A time-dependent decrease in LMS and AChE was noted in exposed earthworms, as well as a concomitant increase in DNA damages (MNi) after 7 and 14 days of exposure. Also, a severe oxidative stress was recorded in animals treated with the different types of compost through an increase in CAT and GST activities, and LPO levels, especially after 14 days of exposure. Therefore, it is necessary to carefully consider these findings for agricultural good practices in terms of plastic mitigation in compost usage, in order to prevent any risk for environment health.

Insights into tolerance mechanisms of earthworms (Eisenia fetida) in copper-contaminated soils by integrating multi-omics analyses.

Earthworms can resist high levels of soil copper (Cu) contamination and play an essential role in absorbing them effectively. However, the molecular mechanisms underlying Cu tolerance in earthworms are poorly understood. To address this research gap, we studied alterations of Eisenia fetida in antioxidant enzymes, gut microbiota, metabolites, and genes under varying levels of Cu exposure soils (0, 67.58, 168.96, 337.92 mg/kg). Our results revealed a reduction in antioxidant enzyme activities across all treatment groups, indicating an adaptive response to alleviate Cu-induced oxidative stress. Analysis of gut microbiota revealed a significant increase in the abundance of bacteria associated with nutrient uptake and Cu2+ excretion under Cu stress. Furthermore, metabolomic analysis discovered an increase in certain metabolites associated with energy metabolism, such as pyruvic acid, L-malic acid, and fumaric acid, as Cu concentration escalated. These results suggested that enhanced energy supply contributes to the elevated tolerance of E. fetida towards Cu. Additionally, transcriptome analysis not only identified crucial detoxification genes (Hsp70, CTSL, GST, CHAC, and GCLC), but also confirmed the critical role of glutathione metabolism as a key pathway in E. fetida Cu detoxification processes. These findings provide a new perspective on the molecular mechanisms of Cu tolerance in earthworms.

Phytometabolites from coral jasmine flower extracts: Toxic effects on Spodoptera litura and enzyme inhibition in nontarget earthworm Eisenia fetida as an alternative approach.

Green pesticides, derived from natural sources, have gained wider attention as an alternative to synthetic pesticides for managing polyphagous pests, such as Spodoptera litura. In this study, the methanolic flower extract of Nyctanthes arbor-tristis (Mx-Na-t) was subjected to chemical screening, and 3-hydroxy-1,2-dimethyl-4(1H)-pyridone (3H-dp) and tyrosol (Ty-ol) were identified as the major derivatives. The toxic effects of Mx-Na-t (500 ppm) were highest in third-instar S. litura larvae (96.4%), while those of 3H-dp and Ty-ol (5 ppm) were highest in second-instar larvae (76.5% and 81.4%, respectively). The growth and development of S. litura larvae and pupae were significantly reduced by all three treatments. Fecundity rates were also reduced by all treatments [from 1020 eggs (control) to 540 eggs by Mx-Na-t treatment, 741 eggs by 3H-dp treatment, and 721 eggs by Ty-ol treatment]. The extract and its active constituents decreased adult emergence and slowed total larval development in a dose-dependent manner. A decrease was noted in the major gut enzymes of young S. litura larvae exposed to Mx-Na-t, 3H-dp, and Ty-ol. Moreover, midgut tissues of fourth-instar larvae were severely damaged by Mx-Na-t (250 ppm), 3H-dp (2.5 ppm), and Ty-ol (2.5 ppm); the treatments induced structural damage to the epithelial cells and gut lumen. The earthworm Eisenia fetida was used to assess nontarget toxicity. Compared with cypermethrin, the phytochemicals exhibited minimal effects on the earthworm's detoxifying enzymes superoxide dismutase and catalase after 14 days of treatment. Moreover, in silico predictions using BeeTox and ProTox-II indicated little or no toxicity of 3H-dp and Ty-ol toward honey bees and other nontarget species.

Surviving in a multistressor world: Gene expression changes in earthworms exposed to heat, desiccation, and chemicals.

An investigation of the effects of anthropogenic stress on terrestrial ecosystems is urgently needed. In this work, we explored how exposure to heat, desiccation, and chemical stress alters the expression of genes that encode heat shock proteins (HSPs), an enzyme that responds to oxidative stress (CAT), hypoxia-related proteins (HIF1 and HYOU), and a DNA repair-related protein (PARP1) in the earthworm Eisenia fetida. Exposure to heat (31°C) for 24 h upregulated HSPs and hypoxia-related genes, suggesting possible acquired thermotolerance. Desiccation showed a similar expression profile; however, the HSP response was activated to a lesser extent. Heat and desiccation activated the small HSP at 24 h, suggesting that they may play a role in adaptation. Simultaneous exposure to endosulfan and temperature for 7 h upregulated all of the evaluated genes, implicating a coordinated response involving multiple biological processes to ensure survival and acclimation. These results highlight the relevance of multistress analysis in terrestrial invertebrates.

Integrated evaluation of the biological response of the earthworm Eisenia fetida using two glyphosate exposure strategies: soil enriched and soils collected from crops in Southeastern Mexico.

Under laboratory conditions, the toxicological effects of pesticides tend to be less variable and realistic than under field conditions, limiting their usefulness in environmental risk assessment. In the current study, the earthworm Eisenia fetida was selected as a bioindicator for assessing glyphosate toxic effects in two different trials to solve this dilemma. In Trial 1, the worms were exposed for 7 and 14 days to concentrations of a commercial glyphosate formulation (1 to 500 mg a.i. kg-1) currently used in the field. In Trial 2, the worms were kept in nine soils collected from different plots with crops for 14 days of exposure. In both experiments, glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and acetylcholinesterase (AChE) activities and contents of lipid peroxidation (LPO) were evaluated. In T1, the glyphosate formulation produced a 40% inhibition of AChE activity and a significant increase in GST, SOD, CAT, and GPx activities and LPO contents in E. fetida on day 7. In T2, higher concentrations of glyphosate were detected in the soils of soybean, papaya, and corn (0.92, 0.87, and 0.85 mg kg-1), which induced a positive correlation between the levels of glyphosate residues with GST, SOD, CAT, GPx, and LPO and a negative correlation with AChE. These findings indicate that crop soils polluted with glyphosate elicited higher oxidative stress than under laboratory conditions, confirmed by IBRv2, PCA, and AHC analyses.