Acute toxicity, biochemical and gene expression responses of the earthworm Eisenia fetida exposed to polycyclic musks

Phytoremediation is a novel and promising approach for the treatment of pollutants. This study did explore the potential of Aster amellus Linn. to decolorize a sulfonated azo dye Remazol Red (RR), a mixture of dyes and a textile effluent. Induction in the activities of lignin peroxidase, tyrosinase, veratryl alcohol oxidase and riboflavin reductase was observed during RR decolorization, suggesting their involvement in the metabolism of RR. UV-Visible absorption spectrum, HPLC and FTIR analysis confirmed the degradation of RR. Four metabolites after the degradation of the dye were identified as 2-[(3-diazenylphenyl) sulfonyl] ethanesulfonate, 4-amino-5-hydroxynaphthalene-2,7-disulfonate, naphthalene-2-sulfonate and 3-(1,3,5-triazin-2-ylamino)benzenesulfonate by using GC/MS. Textile effluent and mixture of dyes showed 47% and 62% decrease respectively in American Dye Manufacturers Institute value. BOD of textile effluent and mixture of dyes were reduced by 75% and 48% respectively, COD of industrial effluent and mixture of dyes was reduced by 60% and 75% and TOC was reduced by 54% and 69% respectively after the treatment by A. amellus for 60 h; this indicated that the plant can be used for cleaning textile effluents. Toxicity study revealed the phytotransformation of RR into non-toxic products.     

Biomarkers responses of the earthworm Eisenia fetida to acetochlor exposure in OECD soil

To examine the potential of a suite of biomarkers as early warning indicators of environmental pollution, sperm count, neutral red retention time (NRRT) and DNA damage were measured in earthworm Eisenia fetida exposed to increasing concentrations of acetochlor in OECD soil. The neutral red retention time of earthworms coelomocytes was sensitive to acetochlor pollution, and decreased significantly when the concentration was more than 10mgkg(-1) after 30 and 60 days of exposure (P<0.05). The reduced neutral red retention time correlated with the soil acetochlor residual. Sperm count decreased significantly at the concentrations of 40 and 80mgkg(-1) after 15 days of exposure (P<0.05). The DNA damage of earthworms coelomocytes increased significantly after 30 days of exposure at the highest concentration (80mgkg(-1); P<0.05). Earthworms were under physiological stress at field dose of acetochlor (10mgkg(-1)). Higher concentrations of acetochlor caused sperm count decrease and DNA damage of earthworms. Such a suite of biomarkers could serve as indicators of the health of the soil environment and to evaluate the toxicity of acetochlor on earthworms or as a means of monitoring soil acetochlor pollution.     

Ecotoxicological responses of the earthworm Eisenia fetida exposed to soil contaminated with HHCB

Although polycyclic musks have been shown to cause lethal and sub-lethal effects on organisms, their biochemical toxicity to earthworms is not well understood. In the current study, we investigated the responses of antioxidant systems and lipid peroxidation after exposing Eisenia fetida to soil contaminated with 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethyl-cyclopenta-γ-2-benzopyran (HHCB). Significant increase in lipid peroxidation level was observed on day 14 at two high concentrations, 50 and 100 mg kg⁻¹. Among antioxidant enzymes, the primary response to chronic HHCB exposure can be attributed to superoxide dismutase (SOD) and catalase (CAT). Of the two enzymes, SOD exhibited more sensitive response to HHCB stress. In addition, these two enzymes could have a combined effect on fighting damage by reactive oxygen species, evidenced by a marked relationship between lipid peroxidation and enzyme activity. On the other hand, dose-dependent inhibition of peroxidase (POD) activity has been observed throughout the test. The results suggest that the variations in investigated parameters of E. fetida could be used as responsive biomarkers for oxidative stress caused by HHCB in a soil environment.     

Ecotoxicological effects on the earthworm Eisenia fetida following exposure to soil contaminated with imidacloprid

Imidacloprid, a neonicotinoid insecticide, has been used widely in agriculture worldwide. The adverse effects of imidacloprid on exposed biota have brought it increasing attention. However, knowledge about the effects of imidacloprid on antioxidant defense systems and digestive systems in the earthworm is vague and not comprehensive. In the present study, the changes in the activity of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), cellulase, reactive oxygen species (ROS), and malondialdehyde (MDA) in the earthworm Eisenia fetida exposed to artificial soil treated with imidacloprid were examined systematically. The results showed that the activity of these biomarkers was closely related to the dose and duration of the exposure to imidacloprid. The activity of SOD was stimulated significantly at doses of 0.66 and 2 mg kg^?1 imidacloprid but markedly inhibited at a dose of 4 mg kg^?1 imidacloprid with prolonged exposure. The activities of CAT and POD increased irregularly at 0.2–4 mg kg^?1 imidacloprid over different exposure times. The level of ROS at a dose of 2 or 4 mg kg^?1 imidacloprid was significantly increased over the entire exposure period. When the concentration of imidacloprid was above 0.66 mg kg^?1, the balance of the activity of the antioxidant enzymes and ROS level was interrupted. The activity of cellulase decreased significantly with prolonged exposure. At the stress of 4 mg kg^?1 imidacloprid, the content of MDA was significantly increased with increasing exposure time. The results of the present study suggest that imidacloprid has a potentially harmful effect on E. fetida and may be helpful for assessment of the risk of imidacloprid to the soil ecosystem environment. However, to obtain more comprehensive toxicity data, it is necessary to investigate the effects of imidacloprid on earthworm using native soils in the future work.     

Bioaccumulation and the soil factors affecting the uptake of arsenic in earthworm, Eisenia fetida

To better understand arsenic (As) bioaccumulation, a soil invertebrate species was exposed to 17 field soils contaminated with arsenic due to mining activity. Earthworms (Eisenia fetida) were kept in the soils for 70 days under laboratory conditions, as body burden increased and failed to reach equilibrium in all soils. After 70 days of exposure, XANES spectra determined that As was biotransformed to a highly reduced form. Uptake kinetics for As was calculated using one compartment model. Stepwise multiple regression suggested that sorbed As in soils are bioaccessible, and uptake is governed by soil properties (iron oxide, sulfate, and dissolved organic carbon) that control As mobility in soils. As in soil solution are highly related to uptake rate except four soils which had relatively high chloride or phosphate. The results imply that uptake of As is through As interaction with soil characteristics as well as direct from the soil solution. Internal validation showed that empirically derived regression equations can be used for predicting As uptake as a function of soil properties within the range of soil properties in the data set.     

Effects of biochar and the earthworm Eisenia fetida on the bioavailability of polycyclic aromatic hydrocarbons and potentially toxic elements

Polycyclic aromatic hydrocarbons (PAHs) and potentially toxic elements (PTEs) were monitored over 56 days in calcareous contaminated-soil amended with either or both biochar and Eisenia fetida. Biochar reduced total (449 to 306 mg kg⁻¹) and bioavailable (cyclodextrin extractable) (276 to 182 mg kg⁻¹) PAHs, PAH concentrations in E. fetida (up to 45%) but also earthworm weight. Earthworms increased PAH bioavailability by >40%. Combined treatment results were similar to the biochar-only treatment. Earthworms increased water soluble Co (3.4 to 29.2 mg kg⁻¹), Cu (60.0 to 120.1 mg kg⁻¹) and Ni (31.7 to 83.0 mg kg⁻¹) but not As, Cd, Pb or Zn; biochar reduced water soluble Cu (60 to 37 mg kg⁻¹). Combined treatment results were similar to the biochar-only treatment but gave a greater reduction in As and Cd mobility. Biochar has contaminated land remediation potential, but its long-term impact on contaminants and soil biota needs to be assessed.     

Ecotoxicological assessment of TiO2 byproducts on the earthworm Eisenia fetida

The increasing production of nanomaterials will in turn increase the release of nanosized byproducts to the environment. The aim of this study was to evaluate the behaviour, uptake and ecotoxicity of TiO₂ byproducts in the earthworm Eisenia fetida. Worms were exposed to suspensions containing 0.1, 1 and 10 mg/L of byproducts for 24 h. Size of TiO₂ byproducts showed aggregation of particles up to 700 μm with laser diffraction. Only worms exposed at 10 mg/L showed bioaccumulation of titanium (ICP-AES), increasing expression of metallothionein and superoxide dismutase mRNA (Real-time PCR) and induction of apoptotic activity (Apostain and TUNEL). TiO₂ byproducts did not induce cytotoxicity on cœlomocytes, but a significant decrease of phagocytosis was observed starting from 0.1 mg/L. In conclusion, bioaccumulation of byproducts and their production of reactive oxygen species could be responsible for the alteration of the antioxidant system in worms.     

Accumulation,speciation and localization of silver nanoparticles in the earthworm Eisenia fetida

Environmental Science and Pollution Research - The use of silver nanoparticles (AgNPs) in agriculture and many consumer products has led to a significant release of Ag in the environment. Although...     

The bioaccumulation of Molybdenum in the earthworm Eisenia andrei: influence of soil properties and ageing

Mo bioaccumulation in the earthworm Eisenia andrei was determined after 28 d exposure in ten different European field soils (pH 4.4-7.8) and an artificial soil, freshly spiked with Na₂MoO₄ at concentrations between 3.2 and 3200 mg Mo kg⁻¹ dry soil. Three field soils were also tested after ageing for 11 months. Earthworm Mo concentrations generally levelled off at high exposure levels but in most soils showed a (nearly) linear increase with increasing soil concentrations in the lower, non-toxic range (below EC10 or NOEC for reproduction effects). Bioaccumulation (BAF) and Bioconcentration factors (BCF) were calculated as the ratio of earthworm concentration to soil and estimated porewater concentrations, respectively. BAFs (0.35-3.44) and BCFs (1.31-276) did not seem much affected by soil concentration, suggesting that earthworms are not capable of regulating their internal Mo concentrations. BAF was best predicted by ammonium oxalate-extractable iron (Fe_ox) and phosphor (P_ox) contents of the soils.     

Comparing the sensitivity of soil invertebrates to pesticides with that of Eisenia fetida

The sole routine testing of the standard earthworm Eisenia fetida for the terrestrial risk assessment of pesticides has been under much debate since other soil invertebrates may be more sensitive than this standard test species. However, the very low availability of laboratory toxicity data for taxa other than E. fetida has greatly hampered sensitivity comparisons. In the present study, the relative tolerance (T_rel) approach was used to enable comparing toxicity thresholds obtained from the US-EPA ECOTOX database, for main terrestrial taxonomic groups and pesticidal types of action (insecticides, fungicides, herbicides, and other) separately. Analyses confirmed previously reported lower and higher sensitivity of collembolans to fungicides and insecticides, respectively. However, various other discrepancies in susceptibility relative to E. fetida were encountered as indicated by species sensitivity distributions and/or calculated 95% confidence intervals of T_rel values. Arachnids and isopods were found to be more sensitive to insecticides, and nematodes to fungicides, as compared to E. fetida. Implications of study findings for the terrestrial risk assessment of pesticides are discussed.     

Influence of activated carbon amendment on the accumulation and elimination of PCBs in the earthworm Eisenia fetida

In this study we investigated the use of activated carbon (AC) as a soil amendment for reducing bioavailability of polychlorinated biphenyls (PCBs) to the earthworm Eisenia fetida. Artificial soil was contaminated with PCBs and used in bioaccumulation experiments fresh or after aging for 19 months. PCB bioaccumulation in earthworms was reduced by 68% when AC was placed as a layer without mixing and by 94% when AC was manually mixed into the soil. Aging of the same AC mixed soil for 19 months resulted in an overall reduction of 99% in PCB biouptake. AC-treated aged soil also showed two orders of magnitude lower equilibrium aqueous concentrations of PCBs compared to untreated aged soils. The findings from this study indicate that application of engineered sorbents like AC to PCB impacted soils may greatly reduce PCB uptake at the base of the terrestrial food chain.     

Genotoxic assessment and optimization of pressmud with the help of exotic earthworm Eisenia fetida

Genotoxicity of pressmud (PM) to Allium cepa was investigated to assess its toxic potential and to elucidate the effect of vermicomposting to reduce its toxicity. The PM produced as a waste by product of the sugar cane industry was mixed with cow dung (CD) at different ratios of 0:100 (V₀), 25:75 (V₂₅), 50:50 (V₅₀), 75:25 (V₇₅) and 100:0 (V₁₀₀) (PM:CD) on a dry weight basis for vermicomposting with Eisenia fetida. Different concentrations of 100 % PM sludge extract (10 %, 20 %, 40 %, 60 %, 80 % and 100 %) and negative control (distilled water) and positive control (maleic hydrazide) were analyzed with A. cepa assay to evaluate frequency of chromosomal aberrations before and after vermicomposting. Percent aberration was greatest (30.8 %) after exposure to 100 % PM extract after 6 h but was reduced to 20.3 % after vermicomposting. Exposure to the extract induced c-mitosis, delayed anaphase, laggards, stickiness and vagrant aberrations. Microscopic examination of root meristem exposed to PM sludge extract showed significant inhibition of mitotic index. Also, the mitotic index decreased with increase in concentration of PM sludge extract. After vermicomposting the mitotic index was increased. However, increasing percentages of PM significantly affected the growth and fecundity of the worms and maximum population size was reached in the 25:75 (PM:CD) feed mixture. Nitrogen, phosphorus, sodium, electrical conductivity (EC) and pH increased from initial feed mixture to the final products (i.e., vermicompost), while organic carbon, C/N ratio and potassium declined in all products of vermicomposting. Scanning electron microscopy (SEM) was recorded to identify the changes in texture with numerous surface irregularities and high porosity that proves to be good vermicompost manure. It could be concluded that vermicomposting could be an important tool to reduce the toxicity of PM as evidenced by the results of genotoxicity.     

Oxidative stress and lipid peroxidation in the earthworm Eisenia fetida induced by low doses of fomesafen

Formesafen is a diphenyl ether herbicide that has adverse effects on non-target animals. However, knowledge about the effect of fomesafen on the antioxidant defense system in earthworms is vague. Thus, it is essential to investigate the effects of fomesafen on the antioxidant defense system in earthworms as a precautionary method. In the present study, earthworms (Eisenia fetida) were exposed to artificial soil treated with a range of concentrations of fomesafen (0, 10, 100, and 500 μg kg^?1) and were collected on the 3rd, 7th, 14th, 21st, and 28th days of exposure. Subsequently, the antioxidant enzyme activities (superoxide dismutase (SOD); catalase (CAT); and guaiacol peroxidase (POD)), reactive oxygen species (ROS) level, and malondialdehyde (MDA) content due to fomesafen treatment were examined in earthworms. Compared with the control, the SOD activity increased on the third and seventh days but decreased on the 14th day due to treatment with 100 and 500 μg kg^?1 of fomesafen. The activities of CAT and POD increased significantly on the third, seventh, and 14th days of exposure. In addition, the ROS level was significantly enhanced throughout the entire experimental period and showed a statistically dose-dependent relationship on the seventh and 14th days. The MDA content markedly increased on the seventh day of exposure; however, obvious changes were not detected at other exposure period. Low doses of fomesafen (≤500 μg kg^?1) may result in oxidative damage and lipid peroxidation in E. fetida by inducing the generation of ROS at short exposure periods (14 days). However, the adverse effects of fomesafen gradually disappear as the cooperation of antioxidant enzymes and exposure time are prolonged. This result may be helpful for further studies on the toxicological mechanisms of fomesafen to earthworms.     

Evaluation of post-Katrina flooded soils for contaminants and toxicity to the soil invertebrates Eisenia fetida and Caenorhabditis elegans

This research evaluated soil samples from a New Orleans neighborhood in the Chalmette, Saint Bernard Parish, that had been inundated by flooding associated with Hurricane Katrina. The goal was to determine if ecological risks persisted from flood waters that had come in contact with hazardous surface chemicals before inundating this low-lying neighborhood for a prolonged period. Research objectives were to establish the presence or absence of volatile organic and heavy metal contaminants, and then asses the toxicity of this soil to Eisenia fetida in a soil exposure assay and Caenorhabditis elegans in a simulated porewater exposure assay. Soil analysis revealed detectable levels of metals and organics in the surface soil at each location. No contaminant was detected in concentrations above human health screening values. Chromium and mercury were detected at levels in excess of typical ecological risk values. Soil extracts revealed concentrations of nitrate, sulfate, and chloride above those from an unflooded background sample. Toxicity testing resulted in no acute effects to either test species, but did show bioaccumulation of arsenic, cadmium, and lead in E. fetida exposed to several samples. The combination of mercury and sulfate provide the potential for mercury methylation should flooding and prolonged inundation occur again.     

Influence of earthworm Eisenia fetida on removal efficiency of N and P in vertical flow constructed wetland

This study investigates biomass, density, photosynthetic activity, and accumulation of nitrogen (N) and phosphorus (P) in three wetland plants (Canna indica, Typha augustifolia, and Phragmites austrail) in response to the introduction of the earthworm Eisenia fetida into a constructed wetland. The removal efficiency of N and P in constructed wetlands were also investigated. Results showed that the photosynthetic rate (P _n), transpiration rate (T _r), and stomatal conductance (S _cond) of C. indica and P. austrail were (p?<?0.05) significantly higher when earthworms were present. The addition of E. fetida increased the N uptake value by above-ground of C. indica, T. augustifolia, and P. australis by 185, 216, and 108 %, respectively; and its P uptake value increased by 300, 355, and 211 %, respectively. Earthworms could enhance photosynthetic activity, density, and biomass of wetland plants in constructed wetland, resulting in the higher N and P uptake. The addition of E. fetida into constructed wetland increased the removal efficiency of TN and TP by 10 and 7 %, respectively. The addition of earthworms into vertical flow constructed wetland increased the removal efficiency of TN and TP, which was related to higher photosynthetic activity and N and P uptake. The addition of earthworms into vertical flow constructed wetland and plant harvests could be the significantly sustainable N and P removal strategy.     

The development of genetically inherited resistance to zinc in laboratory-selected generations of the earthworm Eisenia fetida

The capacity of species to adapt both physiologically and genetically to contaminants may allow populations to persist in polluted environments. Such 'adaptation' can have important implications for risk assessment, since it may mean that prediction based on extrapolation of toxicity studies with na?ve populations may prove invalid for long-term contaminated sites. To investigate the evolution of zinc resistance in Eisenia fetida, worms from a previously unexposed population (parent) were selected and reared over two generations (F1, F2) while exposed to zinc in the laboratory. Relative sensitivities of unexposed and selected generations were then compared by exposing parent, F1 and F2 individuals to zinc in contact filter papers tests. Calculation of effect concentrations from this work indicated differences in sensitivity to zinc for successive generations, with higher toxicity values (LC(50), LC(90), LC(99)) found for the selected worms. The increases in resistance found for F1 and F2 worms were confirmed in a discriminating dose study. In addition to comparing the sensitivities of the parent, F1 and F2 generations for zinc, toxicity tests were also conducted with copper to assess if there was evidence of cross-resistance between the two metals. Results indicated similar increases in resistance to copper to those found for zinc. Mechanisms underlying the increased metal resistance were studied in toxicokinetic experiments. Results indicated no clear trends between the three generations indicating that physiological responses, other than differences in kinetic parameters, are responsible for the increased resistance found in the selected worms.     

Molecular and ultrastructural insights into the earthworm Eisenia fetida of the assessment of ecotoxicity during colistin exposure

Colistin is a peptide antibiotic widely used as a feed additive in animal farming, especially in poultry and swine production, for treatment and prevention of gram-negative bacterial infections, as well as for growth promotion use. When orally ingested, colistin is poorly absorbed and is eliminated almost unaltered by the enteric canal into the environment. Thus, risk of environmental toxicity cannot be ignored. In the present study, we examined the effects of colistin on Heath Shock Protein (HSP) 70, metallothionein (MT) gene expressions, and the ultrastructure of intestinal cells, following treatment of the soil indicator earthworm Eisenia fetida with 10, 20, and 100 mg/kg colistin for 7, 14, and 21 days. The results showed that, compared with the control, the expressions of HSP70 and MT genes changed significantly. Colistin caused up-regulations of HSP70’s expression while inhibited the expression of MT gene. In addition, most mitochondria and endoplasmic reticulum were damaged in the group treated with high concentration. The investigation of gene expressions of HSP70 and MT, as well as pathological alterations in the intestinal cells, may provide important information in terms of ecotoxicity of colistin and can be used as early warning system.     

The effects of the insecticide lambda-Cyhalothrin on the earthworm Eisenia fetida under experimental conditions of tropical and temperate regions

Plant Protection Products can affect soil organisms and thus might have negative impacts on soil functions. Little research has been performed on their impact on tropical soils. Therefore, the effects of the insecticide lambda-Cyhalothrin on earthworms were evaluated in acute and chronic laboratory tests modified for tropical conditions, i.e. at selected temperatures (20 and 28 °C) and with two strains (temperate and tropical) of the compost worm Eisenia fetida. The insecticide was spiked in two natural soils, in OECD artificial soil and a newly developed tropical artificial soil. The effects of lambda-Cyhalothrin did rarely vary in the same soil at tropical (LC50: 68.5-229 mg a.i./kg dry weight (DW); EC50: 54.2-60.2 mg a.i./kg DW) and temperate (LC50: 99.8-140 mg a.i./kg DW; EC50: 37.4-44.5 mg a.i./kg DW) temperatures. In tests with tropical soils and high temperature, effect values differed by up to a factor of ten.     

Assessing the impact of organic and inorganic amendments on the toxicity and bioavailability of a metal-contaminated soil to the earthworm Eisenia andrei

Metal-contaminated soil, from the El Arteal mining district (SE Spain), was remediated with organic (6 % compost) and inorganic amendments (8 % marble sludge) to reduce the mobility of metals and to modify its potential environmental impact. Different measures of metal bioavailability (chemical analysis; survival, growth, reproduction and bioaccumulation in the earthworm Eisenia andrei), were tested in order to evaluate the efficacy of organic and inorganic amendments as immobilizing agents in reducing metal (bio)availability in the contaminated soil. The inorganic amendment reduced water and CaCl₂-extractable concentrations of Cd, Pb, and Zn, while the organic amendment increased these concentrations compared to the untreated soil. The inorganic treatment did not significantly reduce toxicity for the earthworm E. andrei after 28 days exposure. The organic amendment however, made the metal-contaminated soil more toxic to the earthworms, with all earthworms dying in undiluted soil and completely inhibiting reproduction at concentrations higher than 25 %. This may be due to increased available metal concentrations and higher electrical conductivity in the compost-amended soil. No effects of organic and inorganic treatments on metal bioaccumulation in the earthworms were found and metal concentrations in the earthworms increased with increasing total soil concentrations.