Data on the identification of microsatellite markers in Eisenia fetida and Eisenia andrei.

Eisenia fetida and Eisenia andrei are closely related earthworm species that play a crucial part in soil and influence its structure and organic matter cycling. Due to their essential environmental role, they are widely used as model organisms in a vast spectrum of research areas. In this work, we partially sequenced genomes of E. fetida and E. andrei, using Illumina technology (Nano 2 × 250 v2 - MiSeq) and de novo assembly strategy. A total of 3785 and 4258 microsatellite or Simple Sequence Repeat (SSR) markers were identified within E. fetida and E. andrei genomic DNA, respectively. The microsatellite markers will facilitate the analyses of genetic diversity and population genetics studies for the two selected earthworm species and their interspecific hybrids.

Polymorphic microsatellite markers demonstrate hybridization and interspecific gene flow between lumbricid earthworm species, Eisenia andrei and E. fetida.

The lumbricid earthworms Eisenia andrei (Ea) and E. fetida (Ef) have been used as model organisms for studies on hybridization. Previously they have been identified by species specific sequences of the mitochondrial COI gene of maternal origin ('a' or 'f') and the nuclear 28S gene of maternal/paternal origin ('A' or 'F'). In experimental crosses, these hermaphroditic species produce progeny of genotypes Ea (aAA), Ef (fFF) and hybrids (aAF and fFA) originating by self-fertilization or cross-fertilization. To facilitate studies on new aspects of the breeding biology and hybridization of earthworms, polymorphic microsatellite markers were developed based on 12 Ea and 12 Ef specimens and validated on DNA samples extracted from 24 genotyped specimens (aAA, fFF, aAF and fFA) from three laboratory-raised families and 10 of them were applied in the present study. The results indicate that microsatellite markers are valuable tools for tracking interspecific gene flow between these species.

In-site and Ex-site Date Palm Exposure to Heavy Metals Involved Infra-Individual Biomarkers Upregulation.

As a tree of considerable importance in arid regions-date palm, Phoenix dactylifera L. survival in contaminated areas of Sfax city has drawn our attention. Leaf samples of the plants grown in the study area showed high levels of cadmium (Cd), copper (Cu), and chromium (Cr). On the basis of this finding, the cellular mechanisms that explain these metal accumulations were investigated in controlled conditions. After four months of exposure to Cd, Cr, or Cu, high bioconcentration and translocation factor (TF>1) have been shown for date palm plantlets exposed to Cd and low TF values were obtained for plantlets treated with Cr and Cu. Moreover, accumulation of oxidants and antioxidant enzyme activities occurred in exposed roots to Cu and Cd. Secondary metabolites, such as polyphenols and flavonoids, were enhanced in plants exposed at low metal concentrations and declined thereafter. Accumulation of flavonoids in cells may be correlated with the expression of the gene encoding Pdmate5, responsible for the transport of secondary metabolites, especially flavonoids. Other transporter genes responded positively to metal incorporation, especially Pdhma2, but also Pdabcc and Pdnramp6. The latter would be a new candidate gene sensitive to metallic stress in plants. Expressions of gene coding metal chelators were also investigated. Pdpcs1 and Pdmt3 exhibited a strong induction in plants exposed to Cr. These modifications of the expression of some biochemical and molecular based-markers in date palm helped to better understand the ability of the plant to tolerate metals. They could be useful in assessing heavy metal contaminations in polluted soils and may improve accumulation capacity of other plants.

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

The use of silver nanoparticles (AgNPs) in agriculture and many consumer products has led to a significant release of Ag in the environment. Although Ag toxicity in terrestrial organisms has been studied extensively, very little is known about the accumulation capacity and coping mechanisms of organisms in Ag-contaminated soil. In this context, we exposed Eisenia fetida earthworms to artificial OECD soil spiked with a range of concentrations of Ag (AgNPs or AgNO3). The main aims were to (1) identify the location and form of accumulation of Ag in the exposed earthworms and (2) better understand the physiological mechanisms involved in Ag detoxification. The results showed that similar doses of AgNPs or AgNO3 did not have the same effect on E. fetida survival. The two forms of Ag added to soil exhibited substantial differences in speciation at the end of exposure, but the Ag speciation and content of Ag in earthworms were similar, suggesting that biotransformation of Ag occurred. Finally, 3D images of intact earthworms obtained by X-ray micro-computed tomography revealed that Ag accumulated preferentially in the chloragogen tissue, coelomocytes, and nephridial epithelium. Thus, E. fetida bioaccumulates Ag, but a regulation mechanism limits its impact in a very efficient manner. The location of Ag in the organism, the competition between Ag and Cu, and the speciation of internal Ag suggest a link between Ag and the thiol-rich proteins that are widely present in these tissues, most probably metallothioneins, which are key proteins in the sequestration and detoxification of metals.

Medium-term effects of Ag supplied directly or via sewage sludge to an agricultural soil on Eisenia fetida earthworm and soil microbial communities.

The widespread use of silver nanoparticles (AgNPs) in consumer products that release Ag throughout their life cycle has raised potential environmental concerns. AgNPs primarily accumulate in soil through the spreading of sewage sludge (SS). In this study, the effects of direct exposure to AgNPs or indirect exposure via SS contaminated with AgNPs on the earthworm Eisenia fetida and soil microbial communities were compared, through 3 scenarios offering increasing exposure concentrations. The effects of Ag speciation were analyzed by spiking SS with AgNPs or AgNO3 before application to soil. SS treatment strongly impacted Ag speciation due to the formation of Ag2S species that remained sulfided after mixing in the soil. The life traits and expression of lysenin, superoxide dismutase, cd-metallothionein genes in earthworms were not impacted by Ag after 5 weeks of exposure, but direct exposure to Ag without SS led to bioaccumulation of Ag, suggesting transfer in the food chain. Ag exposure led to a decrease in potential carbon respiration only when directly added to the soil. The addition of SS had a greater effect on soil microbial diversity than the form of Ag, and the formation of Ag sulfides in SS reduced the impact of AgNPs on E. fetida and soil microorganisms compared with direct addition.

Species-specific Cd-detoxification mechanisms in lumbricid earthworms Eisenia andrei, Eisenia fetida and their hybrids.

Hermaphroditic lumbricid Eisenia sp. earthworms are ubiquitous and highly resistant to a variety of environmental stressors, including heavy metals. Among the progeny of laboratory mated inter-specific pairs of Eisenia fetida (Ea) and Eisenia andrei (Ef) there are fertile Ha hybrids derived from Ea ova fertilized by Ef spermatozoa and very rare sterile Hf hybrids from Ef ova fertilized by Ea spermatozoa. The aim of the first part of the experiment was to compare the life traits and whole body accumulation of cadmium in adult earthworms from genetically defined Ea, Ef and their hybrids (Ha) exposed for four weeks to commercial soil either unpolluted (control) or cadmium-spiked leading to moderate (M) or high (H) soil pollution (M = 425 and H = 835 mg kg-1 dry soil weight). Such exposure impaired cocoon production but not affected earthworm viability despite the massive Cd bioaccumulation in the whole earthworm bodies reaching at M and H groups 316-454, 203-338, 114-253, and 377-309 mg kg-1 dry body weights of Ea, Ef1, Ef2, and Ha, respectively, surprisingly reaching maximum accumulation quantities in hybrids. The second part of the experiment aimed to investigate cadmium-related defense mechanisms at transcriptomic level in coelomocytes non-invasively extruded from coelomic cavities of the new sets of Ea, Ef, Ha, and Hf earthworms exposed to Cd in microcosms for 0 days (control), 2 days, and 7 days (M = 425 mg kg-1). Expression level of stress-induced Cd-metallothionein (mt) and superoxide dismutase (sod) were gradually up-regulated, while the immune-connected lysenin (lys) was rapidly down-regulated; the expression of glutathione S-transferase (gst) and phytochelatin synthase (pcs) remained unaffected. Mt and sod gene up-regulation and lys gene down-regulation were especially pronounced in Ea-derived hybrids. In sum, capacity of cadmium bioaccumulation and detoxification mechanisms is more efficient in interspecific hybrids than in the pure Ea and Ef species.

Transcriptome assembly and abiotic related gene expression analysis of date palm reveal candidate genes involved in response to cadmium stress.

Date palm in Tunisia is of major economic importance but are also factors of social, environmental and economic stability. An annotated assembly of the transcriptome of cultivar Deglet Nour was reported. RNA was isolated from plant Cd-contaminated leaves, and 37,049 unique Illumina RNA-seq reads were used in a transcriptome assembly. The draft transcriptome assembly consists of 6789 contigs and 17.285 singletons with a means length of 858 bp and 1.042 bp, respectively. The final assembly was functionally annotated using Blast2GO software, allowing the identification of putative genes controlling important agronomic traits. The annotated transcriptome data sets were used to query all known Kyoto Encyclopedia of Genes and Genomes pathways. The most represented molecular functions and biological processes were nucleotide binding and transcription, transport and response to stress and abiotic and biotic stimuli. A prediction of the genes interaction network was proposed by selecting corresponding functionally similar genes from Arabidopsis datasets, downloaded by GeneMANIA version 2.1. Several Cd-responsive genes expression was monitored in in vitro isolated explant of Cd stressed Deglet Nour. Some chelators encoding genes were upregulated confirming in silico findings. Genes encoding HMs transporters in date palm showed expression enhancement more pronounced after 20 days of exposure. P. dactylifera transcriptome provides a valuable resource for future functional analysis of candidate genes involved in metal stress response.

Ecotoxicology of silver nanoparticles and their derivatives introduced in soil with or without sewage sludge: A review of effects on microorganisms, plants and animals.

Silver nanoparticles (AgNPs) are widely incorporated in many products, partly due to their antimicrobial properties. The subsequent discharge of this form of silver into wastewater leads to an accumulation of silver species (AgNPs and derivatives resulting from their chemical transformation), in sewage sludge. As a result of the land application of sewage sludge for agricultural or remediation purposes, soils are the primary receiver media of silver contamination. Research on the long-term impact of AgNPs on the environment is ongoing, and this paper is the first review that summarizes the existing state of scientific knowledge on the potential impact of silver species introduced into the soil via sewage sludge, from microorganisms to earthworms and plants. Silver species can easily enter cells through biological membranes and affect the physiology of organisms, resulting in toxic effects. In soils, exposure to AgNPs may change microbial biomass and diversity, decrease plant growth and inhibit soil invertebrate reproduction. Physiological, biochemical and molecular effects have been documented in various soil organisms and microorganisms. Negative effects on organisms of the dominant form of silver in sewage sludge, silver sulfide (Ag2S), have been observed, although these effects are attenuated compared to the effects of metallic AgNPs. However, silver toxicity is complex to evaluate and much remains unknown about the ecotoxicology of silver species in soils, especially with respect to the possibility of transfer along the trophic chain via accumulation in plant and animal tissues. Critical points related to the hazards associated with the presence of silver species in the environment are described, and important issues concerning the ecotoxicity of sewage sludge applied to soil are discussed to highlight gaps in existing scientific knowledge and essential research directions for improving risk assessment.

Bioaccumulation, antioxidative response, and metallothionein expression in Lupinus luteus L. exposed to heavy metals and silver nanoparticles.

Yellow-lupin (Lupinus luteus L.) was grown on soils contaminated with heavy metals during two parallel studies. In the first one, the soil was contaminated by industrial activities whereas, in the second one, the soil was artificially contaminated with a single metal including Cd, Pb, Zn, Ni (in nitrate form), and Ag (in nitrate and nanoparticles form). The study was performed to assess a plant's response to contamination including its antioxidative response and molecular mechanisms involved in metal detoxification through the expression level of metallothioneins (MTs). Overall, the study provided insights into identification and validation of housekeeping genes (HKG) in L. luteus under exposure to metal stress and showed the effects of selected heavy metals and silver nanoparticles on the expression of metallothioneins, the activity of guaiacol peroxidase (GPX) and bioaccumulation of metals in leaves of L. luteus. As such, HKG validation using BestKeeper, NormFinder, and geNorm software allowed for the selection of four most stable reference genes in a context metal contamination for the selected plant. Moreover, a significant increase in the expression levels of MT was observed in plants grown under heavy metal stress and none on plants grown on 25 mg kg-1 of silver nanoparticles. Also, the GPX activity and MT expression showed statistically significant changes between different conditions and doses which means that they can be used as highly sensitive stress markers for planning the phytoremediation process on a large scale.

The efficiency and economic aspects of phytoremediation technology using Phalaris arundinacea L. and Brassica napus L. combined with compost and nano SiO2 fertilization for the removal of PAH's from soil.

The paper presents an evaluation of efficiency and economic potential of the phytoremediation technology, based on the use of energy crops (P. arundinacea L. and B. Napus L.), combined with the fertilization with compost, supported by the addition of nano SiO2. The experiment was conducted in in-situ conditions, using two experimental blocks, divided according to used plant species. Each block included four types of plots with different fertilization treatments (control plots; treatment with nano SiO2; treatment with compost; treatment with mixture of compost and nano SiO2). During the studied period (three vegetation seasons), a cyclic analysis of 16 PAH's content were conducted. Furthermore, a quantitative determination of biomass production was performed as well as assessment of economic potential of different strategies. Data collected during research, shows that method based on energy crops use, due to the join effect of the dangerous pollutants removal and the production of removable energy resource, can be considered as sustainable and should be recommended for use during heavy polluted soil remediation. Moreover, it should be also noted, that the best results for both PAH's removal efficiency and biomass production, were acquired on plots with P. arundinacea L. cultivation, fertilized with mix of compost and nano SiO2.

Antioxidative enzymes and expression of rbcL gene as tools to monitor heavy metal-related stress in plants.

The aim of the study was to evaluate sensitivity and potential applications of selected biomarkers in phytoremediation under complex heavy metal contamination in Sinapis alba L., Robinia pseudoacacia L. and Lupinus luteus L as a potential tools in effective phytoremediation management. The toxicity assessment was conducted using selected measurement endpoints, both classical and advanced, i.e., germination index, roots length, guaiacol peroxidase activity (GPX), chlorophyll and protein content, the amount of total phenolic compounds (TPC) and level of expression of one of the ribulose-bisphosphate carboxylase genes (rbcL). Moreover, the influence of organic additives: cattle, horse manure, and vermicompost on lowering plant abiotic stress caused by complex heavy metal contamination was studied to assess the possible applications of selected stress markers in large scale phytoremediation planning. The results demonstrated the beneficial effects of selected soil additives on plant development. The 5% difference in the quantity of applied amendment caused statistically significant differences in GPX, TPC, chlorophyll content and expression level of rbcL. Among all endpoints, GPX activity, chlorophyll, and phenolic compounds content, as well as the expression of rbcL, turned out to be the most reliable assays for determination of the type and dosage of selected soil amendments (fertilizers) in the assisted phytoremediation process. Selected markers can be used to achieve the desired level of plant abiotic stress and consequently photosynthesis efficiency and CO2 sequestration. The results showed, that presented assays can be used in different taxonomical groups such as Fabaceae for planning effective phytoremediation process.

Insight into the expression variation of metal-responsive genes in the seedling of date palm (Phoenix dactylifera).

Phytochelatin synthase and metallothionein gene expressions were monitored via qPCR in order to investigate the molecular mechanisms involved in Cd and Cr detoxification in date palm (Phoenix dactylifera). A specific reference gene validation procedure using BestKeeper, NormFinder and geNorm programs allowed selection of the three most stable reference genes in a context of Cd or Cr contamination among six reference gene candidates, namely elongation factor α1, actin, aldehyde dehydrogenase, SAND family, tubulin 6 and TaTa box binding protein. Phytochelatin synthase (pcs) and metallothionein (mt) encoding gene expression were induced from the first days of exposure. At low Cd stress (0.02 mM), genes were still up-regulated until 60th day of exposure. At the highest metal concentrations, however, pcs and mt gene expressions decreased. pcs encoding gene was significantly up-regulated under Cr exposure, and was more responsive to increasing Cr concentration than mt encoding gene. Moreover, exposure to Cd or Cr influenced clearly seed germination and hypocotyls elongation. Thus, the results have proved that both analyzed genes participate in metal detoxification and their expression is regulated at transcriptional level in date palm subjected to Cr and Cd stress. Consequently, variations of expression of mt and pcs genes may serve as early-warning biomarkers of metal stress in this species.

Copper toxicity and date palm (Phoenix dactylifera) seedling tolerance: Monitoring of related biomarkers.

Date palm (Phoenix dactylifera) seeds were exposed to different copper (Cu) solutions to examine plant stress responses. Low Cu concentrations (0.02 and 0.2 mM) caused an increase of seed germination, whereas higher Cu amounts (2 mM) significantly inhibited seed germination, delayed hypocotyl elongation, increased seedling mortality, and reduced the germination index by more than 90%. Metal-related toxicity symptoms appeared after 15 d of 2 mM of Cu exposure. Biochemical activities such as amylase activity and redox balance elements were examined to study the relationship between external Cu amount and internal plant response. The present study showed that amylolytic activity was dose- and time-dependent. Likewise, H2 O2 production increased after exposure to Cu, which was correlated with thiobarbituric acid reactive substance (TBARS) accumulation. Furthermore at low Cu concentrations, superoxide dismutase (SOD) and catalase (CAT) activities increased, suggesting that date palm seed stimulated its metal homeostasis networks. However, the highest cupric ion amounts increased cell oxidant accumulation and reduced enzyme production. Gene expression level measures of P. dactylifera phytochelatin synthase (Pdpcs) and P. dactylifera metallothionein (Pdmt) encoding genes have been carried out to investigate the implication of PdPCS and PdMT proteins in Cu homeostasis and/or its sequestration. Phoenix dactylifera metallothionein induction reached a peak after 30 d of exposure to 0.2 mM of Cu. However, it was down-regulated in plants exposed to higher Cu concentrations. In the same conditions, Pdpcs was overexpressed during 1 mo of exposure before it decreased thereafter. These observations provide a new insight into date palm cell response to Cu, a metal that can be toxic but that is also an essential element. Environ Toxicol Chem 2018;37:797-806. © 2017 SETAC.

Protective role of metallothionein during regeneration in Eisenia andrei exposed to cadmium.

Lumbricid earthworms are often exposed to simultaneous action of various environmental stressors like soil contamination, temperature fluctuation or predators' attacks, which may induce extrusion of coelomocyte-containing coelomic fluid or loss of tail segments. If the injuries are not lethal, renewal of the immune-competent cells and soluble components of coelomic fluid and/or the regeneration of tail segments occurs. The aim of our investigations was to test the hypothesis that exposure of adult earthworms Eisenia andrei to cadmium-polluted soil at room temperature (RT) and/or low temperature (6°C) have adverse effects on restoration of experimentally depleted coelomocytes or on regeneration of amputated posterior segments. Intact control earthworms and their experimental counterparts subjected to electrostimulation-induced coelomocyte depletion or surgical amputation of posterior segments were maintained either in control soil or in soil spiked with cadmium chloride (500mg/kg air-dried soil) at RT or 6°C. Four weeks after the beginning of experiments, cadmium accumulation in earthworm bodies was significantly lower at 6°C than at room temperature. The numbers of restored cells and fluorophore contents were hardly affected by temperature or cadmium. However, cocoon production was reduced by cadmium and completely abolished at 6°C and regeneration of amputated posterior segments was inhibited in cold but was enhanced by cadmium exposure at RT. Independently on the temperature, the 4-week cadmium exposure of adult earthworms was connected with significantly upregulated expression of Cd-metallothionein (but not of catalase, lysenin and phytochelatin) in coelomocytes.

The evaluation of polycyclic aromatic hydrocarbons (PAHs) biodegradation kinetics in soil amended with organic fertilizers and bulking agents.

The aim of this study was to investigate the polycyclic aromatic hydrocarbons (PAHs) biodegradation kinetics in soils fertilized with organic amendments (sewage sludge, compost), bulking agents (mineral sorbent, silicon dioxide in form of nano powder), and novel compositions of those materials. The scope of conducted works includes a cyclic CO2 production measurements and the determinations of PAHs content in soil samples, before and after 3-months of incubation. Obtained results show that the use of both type of organic fertilizers have a positive effect on the PAHs removal from soil. However, the CO2 emission remains higher only in the first stage of the process. The best acquired means in terms of PAHs removal as well as most sustained CO2 production were noted in samples treated with the mixtures of organic fertilizers and bulking agents. In conclusion the addition of structural forming materials to the organic fertilizers was critical for the soil bioremediation efficiency. Therefore, the practical implementation of collected data could find a wide range of applications during the design of new, more effective solutions for the soil bioremediation purposes.

The presence of contaminations in sewage sludge - The current situation.

Sewage sludge/biosolids are by-wastes of municipal and industrial wastewater treatment. As sources of nutrients (C, N, P) they are widely used in intensive farming where large supplementation of organic matter to maintain fertility and enhance crop yields is needed. However, according to the report of European Commission published in 2010, only 39% of produced sewage sludge is recycled into agriculture in the European Union. This situation occurs mainly due to the fact, that the sewage sludge may contain a dangerous volume of different contaminants. For over decades, a great deal of attention has been focused on total concentration of few heavy metals and pathogenic bacteria Salmonella and Escherichia coli. The Sewage Sludge Directive (86/278/EEC) regulates the allowable limits of Zn, Cu, Ni, Pb, Cd, Cr and Hg and pathogens and allows for recovery of sludge on land under defined sanitary and environmentally sound conditions. In this paper, a review on quality of sewage sludge based on the publications after 2010 has been presented. Nowadays there are several papers focusing on new serious threats to human health and ecosystem occurring in sewage sludge - both chemicals (such as toxic trace elements - Se, Ag, Ti; nanoparticles; polyaromatic hydrocarbons; polychlorinated biphenyl; perfluorinated surfactants, polycyclic musks, siloxanes, pesticides, phenols, sweeteners, personal care products, pharmaceuticals, benzotriazoles) and biological traits (Legionella, Yersinia, Escherichia coli O157:H7).

Determination of the performance of vermicomposting process applied to sewage sludge by monitoring of the compost quality and immune responses in three earthworm species: Eisenia fetida, Eisenia andrei and Dendrobaena veneta.

The aim of this study was to assess the effectiveness of vermicomposting process applied on three different sewage sludge (precomposted with grass clippings, sawdust and municipal solid wastes) using three different earthworm species. Selected immune parameters, namely biomarkers of stress and metal body burdens, have been used to biomonitor the vermicomposting process and to assess the impact of contaminants on earthworm's physiology. Biotic and abiotic parameters were also used in order to monitor the process and the quality of the final product. Dendrobaena veneta exhibited much lower resistance in all experimental conditions, as the bodyweight and the total number of circulating immune cells decreased in the most contaminated conditions. All earthworm species accumulated heavy metals as follows Cd>Co>Cu>Zn>Ni>Pb>Cr: Eisenia sp. worms exhibited the highest ability to accumulate several heavy metals. Vermicompost obtained after 45days was acceptable according to agronomic parameters and to compost quality norms in France and Poland.

Sewage sludge disposal strategies for sustainable development.

The main objective of the present review is to compare the existing sewage sludge management solutions in terms of their environmental sustainability. The most commonly used strategies, that include treatment and disposal has been favored within the present state-of-art, considering existing legislation (at European and national level), characterization, ecotoxicology, waste management and actual routs used currently in particular European countries. Selected decision making tools, namely End-of-waste criteria and Life Cycle Assessment has been proposed in order to appropriately assess the possible environmental, economic and technical evaluation of different systems. Therefore, some basic criteria for the best suitable option selection has been described, in the circular economy "from waste to resources" sense. The importance of sewage sludge as a valuable source of matter and energy has been appreciated, as well as a potential risk related to the application of those strategies.

Biochar amendment for integrated composting and vermicomposting of sewage sludge - The effect of biochar on the activity of Eisenia fetida and the obtained vermicompost.

Sewage sludge derived biochar (SSDB) was used as a supplementary material for municipal sewage sludge (SS) and wood chips mixtures (WC) treated by combined composting and vermicomposting. SSDB added to the mixture before composting resulted in significantly higher reproduction rate: on week 4 the number of cocoons increased by 213% when compared to the mixture with no biochar. On week 6 the average number of juveniles increased 11-fold in the mixture with biochar added before composting and 5-fold in the mixtures with biochar added after composting when compared to the mixture with no biochar. Biochar added before composting reduced bioavailability of Cd and Zn to E. fetida. The biochar-added vermicomposts showed good fertilizing properties except for elevated concentrations of Cr. The pH of all vermicomposts was in the range of 5.27-5.61. The obtained vermicomposts can be used as a growing medium for horticultural purposes or as an amendment in calcareous soils.

Vermiremediation of polycyclic aromatic hydrocarbons and heavy metals in sewage sludge composting process.

The main objective of this work was to study the dynamics of the degradation of polycyclic aromatic hydrocarbons (PAHs) during sewage sludge vermicomposting. This eco-biotechnology employing earthworms as natural bioreactors for decomposing of organic matter may be used for vermiremediation of particular pollutants present in various organic matter sources. In this experiment, sewage sludge was mixed with bulking agents and precomposted. Afterward, adult Eisenia andrei specimens were introduced into the process. Total heavy metal and PAHs concentration were measured in composts and earthworms before and after the process. While heavy metal concentrations fluctuated mildly in the substratum, several metals clearly accumulated in the earthworms' bodies. Body Accumulation Factors could be ranked as follows (Cd > Cu > Zn > Ni > Cr > Pb). Interestingly, addition of earthworms into the process has led to the high percentage of PAHs removal and some of the 16 priority PAHs analyzed in this study have been accumulated in earthworms' bodies. Applied conditions did not affect worms' viability but they almost completely inhibited their reproduction.