Metals In Earthworms Research Articles

Comparative Bio-accumulation Potential of Chromium and Cadmium and Their Toxic Effects on Morphology of Earthworm’s Species, Pheretima posthuma

İn this study research was conducted to investigate the accumulative ability and devastating effect of Chromium sulfate (CrSO4) and Cadmium chloride (CdCl2) on the growth of earthworm (EW), Pheretima posthuma. An experimental trial conducted in the Labortory of the Department of Zoology, Wildlife and Fisheries, University of Agriculture Faisalabad. After acclimatization, earthworms reared in jars were divided into five groups with three replicates, Group A controlled, Group B was given 200mg/kg CrSO4, Group C was given 300 mg/kg CrSO4, Group D was given 200mg/kg CdCl2, Group E was given for 300mg/kg CdCl2 treated-soil, respectively. The flame spectrophotometer was used for the quantification of absorption of heavy metals. After 30 days post-exposure, the highest Cr accumulation (8.88±0.07 mg/kg ) was detected in group C while highest accumulation of Cd (7.81±0.09 mg/ kg) was detected in group E. Furthermore, it was noted that these significant accumulations resulted in weight and length reduction. The highest weight reduction (-1.23±0.02g) was observed in group C while the lowest weight reduction (-1.15±0.01g) was observed in group E. Similarly, the highest length reduction (-1.82±0.03cm) was observed in group C while lower length reduction (-1.745±0.24cm) was noted in group E. It was evident that Cr has a more deleterious effect on earthworm growth than Cd. The control showed weight gain (0.26±0.05g) and length gain (1.98±0.27cm). The result of the study were statistically significant (P<0.05). Taken together, it was concluded that earthworm has great potential for bio-accumulation of available heavy metals (Cd and Cr) in the soil and Cr has a higher potential for bio-accumulation than Cd. Although earthworm has a great tendency to survive in a heavy metal environment, their body mass is reduced due to heavy metal toxicity. This study aims to understand the storage of heavy metals in earthworms and to investigate their bioaccumulation ability. This study aims to how heavy metals are stored in earthworms and investigate their capacity for bioaccumulation.

Heavy metal accumulation, biochemical and transcriptomic biomarkers in earthworms Eisenia andrei exposed to industrially contaminated soils from south-eastern Tunisia (Gabes Governorate)

Heavy metal pollution is causing harmful consequences on soil fertility, and earthworms are frequently employed as test organisms to evaluate the ecotoxicity of polluted soils. In this study, Eisenia andrei was exposed for 7 and 14 days to polymetallic contaminated soils collected from an industrial zone in the south-eastern Tunisia. Earthworm growth, heavy metal accumulation, genotoxicity, cytotoxicity, biochemical and transcriptional responses were determined. Results revealed a higher accumulation of heavy metals in earthworms after 14 than 7 days of soil exposure, a reduction in lysosomal membrane stability (LMS), besides an increase in micronuclei frequency (MN). Moreover, earthworm oxidative status was affected in terms of increases in malondialdehyde (MDA) and metallothionein (MTs) content, and enhancement of catalase (CAT) and glutathione-S-transferase (GST) activities. An inhibition of acetylcholinesterase (AChE) activity was also observed in treated earthworms, whereas transcriptional data demonstrated an up-regulation of cat, gst, mt, p21 and topoisomerase genes. Overall, these findings support the use of earthworms as suitable bioindicator species for pollution monitoring and assessment, advance our understanding of the interaction between heavy metals and earthworms, and provide valuable information about the harmful impact of biota exposure to naturally contaminated soils.

Microplastics impact the accumulation of metals in earthworms by changing the gut bacterial communities

Microplastics (MPs) are defined as plastic debris with particle size smaller than 5 mm, which have been frequently detected in environments. In this study, earthworms (Eisenia foetida) were exposed to three different sized polystyrene (PS; 0.1, 10 and 100 μm) at 10 and 100 mg per kg soil for 21 days. We examined the contents of metals (Cu, Zn, Ni and Pb) and gut microbial communities in earthworms exposed to MPs. Results showed that MPs reduced the accumulation of Ni and Pb in earthworms on the 21st day. The composition of gut bacterial communities was altered in earthworms exposed to MPs, especially 10 μm MPs, featuring a higher relative abundance of Proteobacteria (44.5%) and Bacteroidetes (27.1%) than the control group. Additionally, gut microorganisms including genus Paenibacillus and Achromobacter in earthworms were expected to be potential biomarkers for Cu and Zn. The microbial community networks showed that MPs affected bacterial community connections by size effect, and MPs with smaller size increased the complexity of earthworm microbial community networks. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that metabolism was the main difference in the bacterial communities in different treatments, especially carbohydrate metabolism and amino acid metabolism, which was relevant to the survival and growth of bacteria. This study provides insight into the environmental risks of MPs on terrestrial organisms.

Size effects of polystyrene microplastics on the accumulation and toxicity of (semi-)metals in earthworms

Microplastics (MPs) are plastic fragments less than 5 mm, which may have adverse impacts on organisms. In this study, we investigated the impacts and mechanisms of polystyrene MPs (10 μm and 100 μm) and nanoplastics (NPs, 100 nm) with different concentrations (10 mg/kg and 100 mg/kg) in soil on the uptake of metal Cd and semi-metal As in earthworms, Eisenia fetida. MPs facilitated the accumulation of (semi-)metals via damaging the integrity of earthworm intestine, and earthworms accumulated more (semi-)metals in MP treatment groups than NP treatment groups, especially in group of 100 mg/kg of 10 μm MP with concentrations of 1.13 mg/kg and 32.7 mg/kg of Cd and As, respectively. Higher genotoxicity to earthworms was observed for MPs than NPs. Antioxidant enzymes activity and their mRNA gene relative expression levels indicated that MPs with high concentration induced severer damage to earthworms, thus resulting in the increased accumulation of (semi-)metals by earthworms. In addition, proteomic and metabolomic analysis revealed that MPs (100 ppm of 10 μm) disturbed the earthworm immune and metabolic systems, resulting in the highest accumulation of (semi-)metals in earthworms. This study clarifies the influence mechanisms of MPs with different sizes and levels on the accumulation of (semi-)metals by terrestrial invertebrates.

Microplastics Impact the Accumulation of Metals in Earthworms by Changing the Gut Bacterial Communities

Microplastics Impact the Accumulation of Metals in Earthworms by Changing the Gut Bacterial Communities

Ecological risk assessment of heavy metal-contaminated soil using the triad approach

Heavy metal soil-pollution is of concern due to its adverse effects on the ecosystem and human health. However, conventional ecological risk assessments considering only chemical concentration have limitations because of an increased uncertainty in assessing soil-risk. Therefore, this study applied the Triad approach, using a tier system, for assessing site-specific ecological risk in heavy metal-contaminated soil. In addition, risk-level was determined to analyze the necessity of remediation in a contaminated site. Soil samples from six different sites, including heavy metal-contaminated regions (S1–S5) and a control site (C, unpolluted with heavy metals) were collected. Soil properties were assessed based on a tier system to evaluate the ecological risk of each site. The first step tier-1, involved calculating the soil pollution index (SPI) and the second, tier-2, involved assessing site-specific ecological risk based on three lines of evidence (LoEs), viz., chemistry (ChemLoE), ecotoxicology (EcotoxLoE), and ecology (EcoLoE). The total and bioavailable fractions of heavy metals in soil and bio-accumulated heavy metals in earthworms (Eisenia fetida) were measured for ChemLoE. A bioassay involving mortality and weight reduction rates of earthworms, and a root-elongation test of lettuce (Lactuca sativa sp.) was conducted for EcotoxLoE. Soil biological properties, soil basal respiration, and soil enzyme activities, were evaluated for EcoLoE. Final integrated risk (IR) was obtained by combining the risk-values of each LoE, determined using IR values and deviation of each LoE. Although three out of the five sampling sites had SPI > 1 in tier-1, all five sampling sites were subjected to tier-2 to evaluate site-specific ecological risk. In tier-2, ChemLoE (0.53–0.99) was higher than EcotoxLoE (0.05–0.43) and EcoLoE (0.00–0.29), indicating that chemical aspects impacted risk evaluation more than the toxicological and ecological aspects. The final IR ranged between 0.24 and 0.85; risk-level was classified as low (S2, S3, S5), moderate (S1), and high (S4) according to the IR value. Based on the risk-level, site S4 could require remediation for agricultural land-use. The IR of each sampled site varied depending on the impact of heavy metals. Based tier-1 and tier-2 results, a thorough site-specific evaluation is required to understand the adverse effects of heavy metals on soil ecosystems, and the triad approach could be useful for managing heavy metals in soil.

БИОИНДИКАЦИЯ ПОЧВ ГОРОДА МОСКВЫ С ПОМОЩЬЮ ДОЖДЕВЫХ ЧЕРВЕЙ

Relevance. One of the numerous and representative groups of soil animals-bioindicators, represented in all biogeocenoses - are earthworms. A change in the chemistry of the habitat leads to a change in their numbers. The article presents data on the indicator potential of earthworms in soil contamination with heavy metals. Methods and Results. Sampling of soil and earthworms was carried out in 11 sections of the South-Western Administrative District of the city of Moscow, confined to areas of various technogenic loads. The article presents data on the indicator potential of earthworms in soil contamination with heavy metals. A strong feedback was established between the total soil pollution coefficient and the number of worms, a weak feedback with their mass, and a strong direct relationship with the total concentration coefficient of heavy metals in earthworms.

Assessment of trace metal contamination and ecological risk in the forest ecosystem of dexing mining area in northeast Jiangxi Province, China

Samples of soil, earthworms, and tree roots from the forest ecosystem in the Dexing Pb/Zn mining area of Jiangxi Province were collected and the status of trace metal pollution analyzed to assess potential ecological risks. Chemometric and geographic information system methods were used to identify and describe the spatial distributions and the main contamination sources of trace metals. The order of potential ecological risks of trace metals in this area are as follows: cadmium (Cd) > arsenic (As) > copper (Cu) > nickel (Ni) > lead (Pb) > chromium (Cr) > zinc (Zn). Elemental spatial distribution maps showed the existence of zones heavily polluted by trace metals around the mining area. Earthworms and roots of three tree species were also heavily contaminated, with concentrations of trace metals in earthworms much higher than in previous studies. The potential ecological risk index and other soil quality indices indicated that soil had moderate to severe contamination and there were high ecological risks, with Cd making the greatest contribution. Multivariate statistical analyses showed that Cd, As, Cu, Pb, and Zn in soil came from a mining activity source, whereas Ni and Cr primarily originated from a natural source.

Bioaccumulation of heavy metals in earthworms from field contaminated soil in a subtropical area of China

Bioaccumulation factors for heavy metals (Cd, Zn, Cu, and Pb) were examined for selected earthworm species (Metaphire californica, Amynthas homochaetus, Amynthas pecteniferus, and Amynthas heterochaetus) that inhabit metal-polluted soils in a subtropical area (Hunan Province) of South China. The earthworms had high plasticity in inhabiting in situ contaminated areas and showed high uptake of the heavy metals and capability for their accumulation in the tissues. The bioaccumulation factor (BAF) were greatest for cadmium and ranked as follows: Cd (10.6–18.8) >> Zn (1.15–1.75) > Cu (1.01–1.35) > Pb (0.56–0.95). Earthworm species with the similar BAF of heavy metals (p > 0.05) belong to the same ecological group. Within individual groups, Cd, Cu, Zn, Cu, and Pb concentrations in earthworms are consistently predicted by total and extractable fraction (DTPA-extractable) in soil. Our results provide insights into the ecological relationships and variations in the uptake and accumulation of heavy metals in different earthworm species in contaminated soils in China.

Toxic responses of cytochrome P450 sub-enzyme activities to heavy metals exposure in soil and correlation with their bioaccumulation in Eisenia fetida

The dose- and time- dependent responses of cytochrome P450 (CYP) sub-enzyme activities to heavy metals in soil, and the relationships between biomarker responses and metal bioaccumulation in Eisenia fetida were evaluated. Earthworms were exposed to soils spiked with increasing doses of Cd, Cu, Pb or Zn for 21 d. Results demonstrated that EROD and CYP3A4 activities responded significantly with increasing dose and exposure duration. EROD activity significantly (P < 0.05) correlated with CYP3A4 activity exposed to Pb and Cu. The earthworm metal burdens had significant correlation with the total metal concentrations in soil (P < 0.01). The bioaccumulation factor (BAF) decreased with the increasing metal concentration in soil. The order of metal bioavailability to E. fetida was Cd > Zn > Cu > Pb. CYP3A4 activity in Pb-exposed earthworms had a significant correlation with the accumulated metal (P < 0.05). Both EROD and CYP3A4 activities in Cu-exposed worms negatively correlated with BAF (P < 0.05). Based on Discriminant Analysis (DA), CYPs activities were sensitive biomarkers of heavy metals exposure, and we also concluded that different biomarkers with multiple durations could be conducted in the eco-toxicological diagnosis of soil pollution.

Assessment of Trace Element Accumulation by Earthworms in an Orchard Soil Remediation Study Using Soil Amendments

This study assessed potential bioaccumulation of various trace elements in grasses and earthworms as a consequence of soil incorporation of organic amendments for in situ remediation of an orchard field soil contaminated with organochlorine and Pb pesticide residues. In this experiment, four organic amendments of differing total organic carbon content and quality (two types of composted manure, composted biosolids, and biochar) were added to a contaminated orchard field soil, planted with two types of grasses, and tested for their ability to reduce bioaccumulation of organochlorine pesticides and metals in earthworms. The experiment was carried out in 4-L soil microcosms in a controlled environment for 90 days. After 45 days of orchardgrass or perennial ryegrass growth, Lumbricus terrestris L. were introduced to the microcosms and exposed to the experimental soils for 45 days before the experiment was ended. Total trace element concentrations in the added organic amendments were below recommended safe levels and their phytoavailablity and earthworm availability remained low during a 90-day bioremediation study. At the end of the experiment, total tissue concentrations of Cu, Cd, Mn, Pb, and Zn in earthworms and grasses were below recommended safe levels. Total concentrations of Pb in test soil were similar to maximum background levels of Pb recorded in soils in the Eastern USA (100 mg kg−1 d.w.) because of previous application of orchard pesticides. Addition of aged dairy manure compost and presence of grasses was effective in reducing the accumulation of soil-derived Pb in earthworms, thus reducing the risk of soil Pb entry into wildlife food chains.

A Hierarchic Method for Studying the Distribution of Phenanthrene in Eisenia fetida

The distribution of heavy metals in earthworms has been widely studied, highlighting the importance of the fate of these metals. However, little information is available on the distribution of hydrophobic organic contaminants (HOCs) within earthworms. The aim of this study was to propose a hierarchic method to study the distribution of phenanthrene (PHE), a typical HOC, in Eisenia fetida at several levels: sub-organism (pre-clitellum, clitellum and post-clitellum), tissue (body wall, gut and body fluid) and subcellular (intracellular and extracellular fractions). Earthworms were incubated in the soils amended with low (LC, 10 mg kg−1) and high concentrations (HC, 50 mg kg−1) of PHE and sampled at different time intervals. At the sub-organism level, the distribution of PHE was homogeneous among the sub-organism fractions in the LC treatment but heterogeneous in the HC treatment and gradually reached the following form of post-clitellum ≈ clitellum > pre-clitellum. The uptake and elimination kinetics of PHE in the sub-organism were well described by a one-compartment model. At the tissue level, the concentration of PHE followed the order of gut > body fluid > body wall; while at the subcellular level, the concentration of PHE in the extracellular fraction was 1.23 to 4.68 times higher than that in the intracellular fraction. Therefore, the simple circulatory system of earthworms may account for the PHE distribution at the sub-organism level. Partition pathways (passive diffusion) of PHE between the body wall, body fluid and gut as well as the processes of PHE entrance into the inner cellular compartment via passive diffusion were experimentally supported.

Soil properties and metal accumulation by earthworms in the Siena urban area (Italy)

This paper reports the results of a study focused on the metal (Cd, Co, Cr, Cu, Ni, Pb, Sb, U and Zn) distribution in soils and uptake and accumulation by earthworms Nicodrilus caliginosus (Savigny) from urban, peri-urban, green-urban and non-urban zones of Siena municipality (central Italy). The main goal was to define the influence of soil properties and metal soil contents on the uptake of these contaminants by earthworms. Data indicated that Cd, Cu, Pb, Sb and Zn soil contents increased in the following order: non-urban<green-urban<peri-urban<urban soils, suggesting that vehicular traffic affects the distribution of these metals. Pb and Sb were the main soil contaminants and their highest enrichments were found in urban sites where stop-and-go traffic occurs. Concentrations of these traffic-related metals in earthworms showed a distribution pattern similar to that in soil, suggesting that soil contamination influenced the uptake of Cd, Cu, Pb, Sb and Zn by N. caliginosus. There were significant positive correlations between Cd, Pb and Sb earthworm concentrations and their soil contents. The lack of correlation for Cu and Zn could be due to the physiological regulation of these elements by earthworms. Statistical analysis pointed out that the uptake and accumulation of Cd, Cu, Pb, Sb and Zn by earthworms were affected by some soil physicochemical properties such as the organic carbon and carbonate contents that are able to rule the bioavailability of metals in soils.

Effects of gypsum on trace metals in soils and earthworms.

Mined gypsum has been beneficially used for many years as an agricultural amendment. A large amount of flue gas desulfurization (FGD) gypsum is produced by removal of SO from flue gas streams when fuels with high S content are burned. The FGD gypsum, similar to mined gypsum, can enhance crop production. However, information is lacking concerning the potential environmental impacts of trace metals, especially Hg, in the FGD gypsum. Flue gas desulfurization and mined gypsums were evaluated to determine their ability to affect concentrations of Hg and other trace elements in soils and earthworms. The study was conducted at four field sites across the United States (Ohio, Indiana, Alabama, and Wisconsin). The application rates of gypsums ranged from 2.2 Mg ha in Indiana to 20 Mg ha in Ohio and Alabama. These rates are 2 to 10 times higher than typically recommended. The lengths of time from gypsum application to soil and earthworm sampling were 5 and 18 mo in Ohio, 6 mo in Indiana, 11 mo in Alabama, and 4 mo in Wisconsin. Earthworm numbers and biomass were decreased by FGD and mined gypsums in Ohio. Among all the elements examined, Hg was slightly increased in soils and earthworms in the FGD gypsum treatments compared with the control and the mined gypsum treatments. The differences were not statistically significant except for the Hg concentration in the soil at the Wisconsin site. Selenium in earthworms in the FGD gypsum treatments was statistically higher than in the controls but not higher than in the mined gypsum treatments at the Indiana and Wisconsin sites. Bioaccumulation factors for nondepurated earthworms were statistically similar or lower for the FGD gypsum treatments compared with the controls for all elements. Use of FGD gypsum at normal recommended agricultural rates seems not to have a significant impact on concentrations of trace metals in earthworms and soils.

Vermiremoval of heavy metal in sewage sludge by utilising Lumbricus rubellus

Experiments were conducted to remove heavy metals (Cr, Cd, Pb, Cu and Zn) from urban sewage sludge (SS) amended with spent mushroom compost (SMC) using worms, Lumbricus rubellus, for 105 days, after 21 days of pre-composting. Five combinations of SS/SMC treatments were prepared in triplicate along with a control for each treatment in microcosms. Analysis of the earthworms' multiplication and growth and laboratory analysis were conducted during the tenth and fifteenth week of vermicomposting. Our result showed that the final biomass of earthworms (mg) and final number of earthworms showed significant differences between treatments i.e. F=554.70, P=0.00 and F=729.10, P=0.00 respectively. The heavy metals Cr, Cd and Pb contained in vermicompost were lower than initial concentrations, with 90–98.7percent removal on week ten. However, concentrations of Cu and Zn, that are considered as micronutrients, were higher than initial concentrations, but they were 10–200-fold lower than the EU and USA biosolid compost limits and Malaysian Recommended Site Screening Levels for Contaminated Land (SSLs). An increment of heavy metals were recorded in vermicompost for all treatments on week fifteen compared to week ten, while concentration of heavy metals in earthworms' tissue were lower compared to vermicompost. Hence, it is suggested that earthworms begin to discharge heavy metals into their surroundings and it was evident that the earthworms' heavy metals excretion period was within the interval of ten to fifteen weeks.

Biosynthesis of luminescent quantum dots in an earthworm

The synthesis of designer solid-state materials by living organisms is an emerging field in bio-nanotechnology. Key examples include the use of engineered viruses as templates for cobalt oxide (Co(3)O(4)) particles, superparamagnetic cobalt-platinum alloy nanowires and gold-cobalt oxide nanowires for photovoltaic and battery-related applications. Here, we show that the earthworm's metal detoxification pathway can be exploited to produce luminescent, water-soluble semiconductor cadmium telluride (CdTe) quantum dots that emit in the green region of the visible spectrum when excited in the ultraviolet region. Standard wild-type Lumbricus rubellus earthworms were exposed to soil spiked with CdCl(2) and Na(2)TeO(3) salts for 11 days. Luminescent quantum dots were isolated from chloragogenous tissues surrounding the gut of the worm, and were successfully used in live-cell imaging. The addition of polyethylene glycol on the surface of the quantum dots allowed for non-targeted, fluid-phase uptake by macrophage cells.

Short-term changes of metal availability in soil. II: The influence of earthworm activity

This study aimed to evaluate short-term earthworm-induced changes in the availability of metals applied to soil directly (metal-spiked) or via an organic matrix (sludge-amended). A laboratory experiment was performed using destructive sampling of microcosms filled with agricultural soil. A concentration gradient of industrial sludge contaminated predominantly with Cr, Cu, Ni, and Zn, and a soil freshly spiked with the same metal concentrations were applied on top of the soil columns. Individuals of Dendrobaena veneta (mimicking a realistic density of 500 earthworms per m 2) were introduced in half of the replicates of each treatment. Total and 0.01 M CaCl 2 extractable metal concentrations were measured in soil after 0, 3, 6 and 12 weeks and metal concentrations in earthworms and percolates were measured after 3, 6, and 12 weeks. Earthworm activity did not affect metal availability of any treatment over time, but Ni and Cu concentrations in D. veneta were higher at the highest treatment levels. Earthworm Zn concentrations were similar in all treatments while Cr concentrations increased with increasing soil total metal content only for sludge treatments. Existing relationships of earthworm metal concentrations with total metal content in soil, taken from the literature, were not able to predict the metal levels measured in D. veneta. Results demonstrated that although over 12 weeks earthworm activity did not affect metal availability in soil, their burrowing activities did influence the metal concentrations of percolates over time.

Biokinetic modeling of heavy metals in earthworms

Studies concerning bioaccumulation kinetics and bioconcentration factor (BCF) of heavy metals like zinc (Zn), lead (Pb), chromium (Cr), cadmium (Cd), and copper (Cu) in earthworm Eudrilus eugeniae tissues including integument, gizzard, clitellum, and head region were undertaken. Calculated BCF, predicted K ow, and predicted K oc showed a significant correlation between heavy metals in different earthworm tissues, in substrate spiked with heavy metals. The regression coefficient (r 2) between heavy metal uptake concentration and exposure time varied between 0.73 and 0.99, indicating significant correlation. The K oc was a maximum of 13.9016 in case of Cu and integument at an exposure time of 100 days and a minimum of 0.1114 in case of Cr with respect to head at the same exposure time. Earthworms accumulated heavy metals following chronic exposure to municipal solid waste containing heavy metals. BCF and uptake rate kinetics of heavy metals were calculated and showed increased values in head tissue followed by integument.

Body metal concentrations and glycogen reserves in earthworms (Dendrobaena octaedra) from contaminated and uncontaminated forest soil

Stress originating from toxicants such as heavy metals can induce compensatory changes in the energy metabolism of organisms due to increased energy expenses associated with detoxification and excretion processes. These energy expenses may be reflected in the available energy reserves such as glycogen. In a field study the earthworm, Dendrobaena octaedra, was collected from polluted areas, and from unpolluted reference areas. If present in the environment, cadmium, lead and copper accumulated to high concentrations in D. octaedra. In contrast, other toxic metals such as aluminium, nickel and zinc appeared to be regulated and kept at low internal concentrations compared to soil concentrations. Lead, cadmium and copper accumulation did not correlate with glycogen reserves of individual worms. In contrast, aluminium, nickel and zinc were negatively correlated with glycogen reserves. These results suggest that coping with different metals in earthworms is associated with differential energy demands depending on the associated detoxification strategy.

Sewage sludge applied to agricultural soil: Ecotoxicological effects on representative soil organisms

Application of sewage sludge to agricultural lands is a current practice in EU. European legislation permits its use when concentrations of metals in soil do not increase above the maximum permissible limits. In order to assess the fate and the effects on representative soil organisms of sewage sludge amendments on agricultural lands, a soil microcosm (multi-species soil system—MS 3) experiment was performed. The MS 3 columns were filled with spiked soil at three different doses: 30, 60 and 120 t ha −1 fresh wt. Seed plants ( Triticum aestivum, Vicia sativa and Brassica rapa) and earthworms ( Eisenia fetida) were introduced into the systems. After a 21-d exposure period, a statistically significant increase for Cd, Cu, Zn and Hg concentrations was found for the soils treated with the highest application rate. Dose-related increase was observed for nickel concentrations in leachates. Plants and earthworm metal body burden offer much more information than metal concentrations and help to understand the potential for metal accumulation. Bioaccumulation factor (BAF plant−soil) presented a different behavior among species and large differences for BAF earthworm−soil, from control or sewage-amended soil, for Cd and Hg were found. B. rapa seed germination was reduced. Statistically significant decrease in fresh biomass was observed for T. aestivum and V. sativa at the highest application rate, whereas B. rapa biomass decreased at any application rate. Enzymatic activities (dehydrogenase and phosphatase) as well as respiration rate on soil microorganisms were enlarged.