Transcriptional expression levels and biochemical markers of oxidative stress in the earthworm Eisenia andrei after exposure to 2,4-dichlorophenoxyacetic acid (2,4-D).

This study investigated the stress response of earthworms (Eisenia andrei) to exposure to a commonly used herbicide, 2,4 dichloro-phenoxy-acetic acid (2,4-D). We evaluated both stress biomarkers and the transcriptional expression levels and activity of three enzymes involved in oxidative stress responses. Earthworms were exposed to three sublethal concentration of 2,4-D (3.5, 7, and 14 mg kg(-1)) for 7 and 14 days. Exposure to 7 and 14 mg kg(-1) 2,4-D significantly reduced both worm body weight and lysosomal membrane stability (LMS); the latter is a sensitive stress biomarker in coelomocytes. Exposure to 2,4-D caused a pronounced increase in the accumulation of malonedialdehyde (MDA), a marker of oxidative stress, and significantly increased the activity of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD),and glutathione-S-transferase (GST). Compared to expression in controls, the expression levels of the sod, cat, and gst genes increased in worms exposed to all three 2,4-D doses for 7 days. However, after 14 days of exposure, only the expression of the gst gene remained higher than controls. These data provide new insights into the cytotoxicity of 2,4-D in the earthworm E. andrei and should be carefully considered in view of the biological effects of herbicides in soils organisms.

Proteomic analysis in caged Mediterranean crab (Carcinus maenas) and chemical contaminant exposure in Téboulba Harbour, Tunisia.

This study uses proteomics approach to assess the toxic effects of contaminants in the Mediterranean crab (Carcinus maenas) after transplantation into Téboulba fishing harbour. High levels of aliphatic and aromatic hydrocarbons were detected in sediments. Although their effects on vertebrates are well described, little is known about their early biological effects in marine invertebrates under realistic conditions. Protein expression profiles of crabs caged for 15, 30 and 60 days were compared to unexposed animals. Nineteen proteins with significant expression differences were identified by capLC-µESI-IT MS/MS and homology search on databases. Differentially expressed proteins were assigned to five different categories of biological function including: (1) chitin catabolism, (2) proteolysis, (3) exoskeleton biosynthesis, (4) protein folding and stress response, and (5) transport. The proteins showing major expression changes in C. maenas after different caging times may be considered as novel molecular biomarkers for effectively biomonitoring aquatic environment contamination.

Biochemical responses in seabream (Sparus aurata) caged in-field or exposed to benzo(a)pyrene and paraquat. Characterization of glutathione S-transferases.

Gilthead seabream (Sparus aurata) specimens were caged in-field at the Téboulba harbour or exposed to benzo(a)pyrene [B(a)P] or to paraquat [PQ] plus B(a)P, and several biochemical biomarker responses were investigated. Antioxidant enzymes, such as glutathione peroxidase, catalase and glutathione reductase, significantly increased in the in-field and B(a)P+PQ exposures, but were only moderately affected by B(a)P alone. Glucose-6-phosphate and 6-phosphogluconate dehydrogenases significantly diminished after in-field exposure. Different responses with biotransformation enzymes were observed: the P4501A-associated EROD activity was highly induced in response to B(a)P and B(a)P+PQ exposures, while total activity of the glutathione S-transferase (GST) was similar to control. However, after purification of the GST proteins by affinity chromatograpy and analysis by two-dimensional electrophoresis, nineteen highly reproducible isoforms were resolved. In addition, some of reproducible isoforms showed different and specific expression patterns in response to contaminants. Thus, proteomic analysis of the purified GST subunits is a reliable tool for ecotoxicological research, useful in polluted marine ecosystem as an effective biomarker of contamination.

Biochemical responses and metals levels in Ruditapes decussatus after exposure to treated municipal effluents.

This study assessed the responses of biochemical biomarkers and metals levels in Ruditapes decussatus exposed to the increasing concentrations of treated municipal effluents (TME) discharged into the Tunisian coastal area. Clams were exposed to 0%, 1%, 3% and 10% for 7 and 14 day and the following biochemical responses were measured: (1) catalase activity and lipid peroxidation levels (TBARS) as oxidative stress biomarkers, (2) gluthathione S-transferase (GST) activity as a phase II conjugation enzyme; (3) cholinesterase activity (ChE) as biomarker of neurotoxicity, and (4) metallothioneins as a proteins highly induced by heavy metals. A significant uptake of Cu, Cd and Zn in digestive gland and serious biochemical alterations were observed. Thus, exposure of clams to croissant concentration of TME have the potential to increase the oxidative stress biomarkers (TBARS, CAT activity) and MT levels; and decrease ChE activity in both gills and digestive gland. Current experimental results suggest that CAT, GST, ChE activities and MT and TBARs levels in gills and digestive gland of clam R. decussatus are sensitive and suitable responses for assessing the effects of anthropogenic contaminants on the aquatic ecosystems, particularly effluent complex mixtures.

Increased temperatures affect oxidative stress markers and detoxification response to benzo[a]pyrene exposure in mussel Mytilus galloprovincialis.

The present research work was designed to study mussel's (Mytilus galloprovincialis) digestive gland biotransformation and detoxification responses to benzo[a]pyrene (B[a]P) exposure along with heat stress. Mussels were exposed to a sublethal dose of B[a]P [75 nM (19 µg/L/animal)] + temperature gradient (18, 20, 22, 24 and 26 °C) for 7 days. B[a]P hydroxylase (BPH) and glutathione-S-transferase (GST) activities were assessed in digestive gland tissues as phase I and phase II biotransformation parameters. Catalase (CAT) activity and malonedialdehyde (MDA) were measured as potential biomarkers of oxidative stress and lipid peroxidation. The cholinergic system was evaluated using acetylcholinesterase (AChE) activity. DNA damage was assessed using micronuclei (MN) test. BPH and GST activities showed a decreasing trend in B[a]P-exposed animals at 24 and 26 °C. CAT activity showed a bell-shaped response in B[a]P-exposed and in heat-stressed organisms at a maximum temperature of 22 °C. AChE activity was significantly inhibited in response to B[a]P being more pronounced at a temperature of 26 °C. MN in digestive gland cells suggest that B[a]P exposure induced significant DNA alteration with a maximum response in organisms coexposed to B[a]P and a temperature of 26 °C. Biomarker data are further discussed in relation B[a]P accumulation in mussels digestive gland. These data should be carefully considered in view of the biological effects of organic pollutants, particularly in organisms under the challenging effects of extreme temperature fluctuations.

Mixture toxicity assessment of nickel and chlorpyrifos in the sea bass Dicentrarchus labrax.

The present research work was designed to study Dicentrarchus labrax biotransformation and detoxification responses to acute exposure to nickel (Ni) and chlorpyrifos (CHP). Sexually immature sea bass were treated by intraperitoneal injection of nickel chloride (500 µg kg⁻¹), chlorpyrifos (10 mg kg⁻¹), and their binary mixture for 1, 3, and 7 days. Ni and CHP accumulation was quantified in liver after the exposure periods. The following biological responses were measured: (1) NADPH cytochrome P450 reductase (NCR) activity, as phase I biotransformation parameter; (2) gluthathione S-transferase (GST) activity as a phase II conjugation enzyme, acetylcholinesterase activity, and metallothionein (MT) content. Ni bioaccumulation in the liver resulted in an increasing uptake up to 15.48 µg g⁻¹ wet weight (Ni-treated animals) and 16.73 µg g⁻¹ wet weight (mixture-treated animals) after 7 days of exposure. CHP accumulation showed a distinct pattern in animals exposed to the mixture of chemicals in comparison with CHP-treated animals. NCR activity exhibited a marked activation in CHP and mixture-treated animals. GST activity was significantly increased starting from 1 day exposure in CHP-treated animals and after 3 days in Ni-treated animals. MT accumulation increased in all conditions, with a marked synergetic effect after 7 days of exposure. These data should be carefully considered in view of the biological effects of mixture pollutants, particularly in fish farming conditions.

Integrated assessment of biochemical responses in Mediterranean crab (Carcinus maenas) collected from Monastir Bay, Tunisia.

The biochemical response of Mediteranean Crab (Carcinus maenas) collected at five stations of Monastir Bay and from Kuriat station as control was studied using a set of complementary biomarkers. The catalase, glutathione S-transferase, lactate dehydrogenase, acetycholinesterase activities; and metallothionein and malonediladehyde levels in gills were evaluated. Results revealed differences among sites in relation to each specific biomarker. Hence, a suite of biomarkers can be used to discriminate sampling sites according to types of pollution, reflecting differing conditions of anthropogenic impact. Based on Integrated Biomarker Response, the highest values and critical biochemical alteration were observed at Khniss and Ksibat in response to urban and industrial discharges and the lowest IBR value was found at reference site. The current study has shown clearly that a biomarker-based index is usefulness tool in the monitoring Tunisian coast using C. maenas as sentinel specie. Further studies in progress to investigate the seasonal variations of IBR levels and its relationship to pollutants concentrations in the sediment, gills and digestive gland of Carcinus maenas from Monastir Bay.

Uptake and biochemical responses of mussels Mytilus galloprovincialis exposed to sublethal nickel concentrations.

In the present study, mussel (Mytilus galloprovincialis) digestive gland oxidative stress biomarkers and detoxification responses to acute exposure to nickel (Ni) were investigated. Mussels were exposed to two sublethal concentrations of Ni (135 µg/L per animal (2.5 µM) and 770 µg/L per animal (13 µM)) for 24, 48, 72, 96 h and 8 days. Following biological responses were measured: (1) glutathione S-transferase (GST) activity as a phase II conjugation enzyme, (2) catalase activity as antioxidant response, (3) malondialdehyde accumulation (MDA) as lipid peroxydation marker and metallothionein as specific response to metals exposure. The cholinergic system was evaluated using the acetylcholinesterase activity (AChE). Moreover, Ni uptakes during the exposure periods were assessed and the uptake rate constant determined. A correlation matrix (CM) between the investigated biomarkers and a principal component analysis (PCA) were achieved for the two tested concentrations. The Ni-uptake constant was higher in animals exposed to the lowest concentration. The CM and the PCA showed a time-dependent effect of the Ni exposure on the investigated biomarkers being more pronounced in animals exposed to the highest Ni concentration. While AChE showed a significant increase after 48 h and a further return to control values in the lowest concentration, it was drastically maintained inhibited in the highest concentration. Our data provided clues about the occurrence of different toxicokinetics and toxicodynamics of two Ni sublethal concentrations in an ecologically relevant organism.

Monitoring pollution in Tunisian coasts using a scale of classification based on biochemical markers in worms Nereis (Hediste) diversicolor.

This paper aims to assess the marine environment quality along the Tunisian coasts using a statistical approach based on biomarkers response in the polychaete worms Nereis (Hediste) diversicolor. Worms were collected from six sites: Bizerta Lagoon, Gargour, Nakta, Mahres, Skhira and from Teboulba considered as a reference site. The biomarkers selected in this work were (1) the activities of cytochrome P450-dependent NADPH cytochrome c reductase (NADPH red) as phase I enzyme, (2) glutathione S-transferase as phase II enzyme and (3) the acetylcholinesterase activity as neurotoxicity marker. Oxidative stress was evaluated using catalase activity and malondialdehyde accumulation. For each biomarker, a discriminatory factor was calculated and a response index was allocated. For each site, a multi-marker pollution index was calculated as the sum of the response index of each of the five more discriminating biomarkers. The results show differences between sites compared with the reference samples. The multi-marker approach confirms that worms from Bizerta and Mahress have been submitted to highly polluted environment. Mahress shows the highest multi-marker pollution index, indicating a highly contamination status.

Multimarker approach analysis in common carp Cyprinus carpio sampled from three freshwater sites.

The aim of this study is to assess the response of a multimarker approach in common carp Cyprinus carpio sampled from three Tunisian dam lakes selected according to different environmental and ecological characteristics. Glutathione-S-transferase (GST) activity was analyzed in carp liver and used as a phase II detoxification enzyme, hepatic metallothionein content (MTs) was used as a metallic stress indicator, and cholinesterase activities were analyzed in muscle and brain and used as neurotoxicity biomarker. Micronucleus frequency test (MN) as a genotoxicity marker. GST and MT levels showed an increase in fish from the Bir Mcherga site and a decrease in Sidi Saâd site with respect to fish from Nebhana site. Results showed a strong inhibition of cholinesterase activities in fish from Bir Mcherga and Sidi Saâd sites compared to Nebhana site. Relatively high level of MN is reported specially in fish blood from the Bir Mcherga site.

Evaluation of enzymatic biomarkers and lipoperoxidation level in Hediste diversicolor exposed to copper and benzo[a]pyrene.

This study aims to evaluate the effects of exposure to copper, benzo[a]pyrene, and to their mixture on enzymatic and lipid peroxidation biomarkers in Hediste diversicolor. Worms were submitted to 1 microM of both single compounds and to their mixture during a period of test of 12, 24, 36, and 48 h. The biomarkers selected in this work were the activities of cytochrome P450-dependent NADPH cytochrome c reductase (NADPH red) as phase I enzyme, glutathione-S-transferase (GST) as phase II enzyme, and the acetylcholinesterase (AChE) activity as neurotoxicity marker. Oxidative stress was evaluated using catalase activity (CAT) and malondialdehyde accumulation (MDA). The NADPH red activity was not significantly affected by copper exposure; it shows a drastic increase in both B[a]P and mixture-exposed organisms. GST activities were significant in B[a]P-exposed worms only after 36 h, and in animals exposed to the mixture after 12 and 48 h. The ACHE activity was inhibited only in B[a]P-exposed worms.

Seasonal variation of oxidative stress biomarkers in clams Ruditapes decussatus sampled from Tunisian coastal areas.

Seven (7) sites from to the Tunisian coastal area were monitored through four seasons using several oxidative stress biomarkers including lipid peroxidation. Catalase (CAT) specific activity was determined in clam Ruditapes decussatus digestive gland. Lipid peroxidation was assessed in the same tissues using malondialdehyde (MDA) concentration, neutral lipid (NL) and lipofuscin accumulation. Results show that catalase specific activity varies between sites and fluctuates in time. The highest CAT activities were recorded in sites from Bizerta Lagoon and sites neighbouring Sfax city. Malondialdehyde accumulation in digestive gland cells seems to be very pronounced in animals sampled in summer and autumn. Digestive gland sections of clams sampled from reported heavy metal polluted sites exhibited higher volume density of residual bodies. Our results showed that the neutral lipids and lipofuscin content in clam's digestive gland are more sensitive to general stress as compared to other biomarkers, and could be used successfully in clams as early warning tools in field biomonitoring programs.

Acute effects of chlorpyryphos-ethyl and secondary treated effluents on acetylcholinesterase and butyrylcholinesterase activities in Carcinus maenas.

The acute effects of commercial formulation of chlorpyrifos-ethyl (Dursban) and the secondary treated industrial/urban effluent (STIUE) exposure on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities in hepatopancreas and gills of Mediterranean crab Carcinus maenas were investigated. After 2 d of exposure to chlorpyriphos-ethyl, the AChE activity was inhibited in both organs at concentrations of 3.12 and 7.82 microg/L, whereas the BuChE was inhibited only at higher concentration 7.82 microg/L of commercial preparation Dursban. The exposure of crabs to Dursban (3.12 microg/L) showed a significant decrement of AChE activity at 24 and 48 h, whereas the BuChE was inhibited only after 24 h and no inhibition for both enzymes was observed after 72 h. Moreover, a significant repression of AChE activity was observed in both organs of C. maenas exposed to 5% of STIUE. Our experiments indicated that the measurement of AChE activity in gills and hepatopancreas of C. maenas would be useful biomarker of organophosphorous (OP) and of neurotoxic effects of STIUE in Tunisia.

Metallothionein induction by Cu, Cd and Hg in Dicentrarchus labrax liver: assessment by RP-HPLC with fluorescence detection and spectrophotometry.

Metallothionein was quantified in sea bass Dicentrarchus labrax intraperitoneally (i.p.) injected with different Cu, Cd and Hg doses (50-250 microg kg(-1) wet wt) after 48 h exposure. A distinct peak with 16.8 min retention time was obtained by reversed-phase high performance liquid chromatography coupled to fluorescence detection (RP-HPLC-FD) with the three metals. Total metallothionein levels assayed in unheated liver extracts by RP-HPLC-FD were significantly higher (1.3-1.95-fold) than those obtained by the well-established spectrophotometric method. In the RP-HPLC-FD method, metallothionein increased linearly with Cu and Hg doses, being saturated beyond 100 mug kg(-1) Cd. Maximum induction was obtained at 100 microg kg(-1) Cd (5.3-fold), and 250 microg kg(-1) Cu or Hg (8- and 5.1-fold, respectively). At low doses no metallothionein induction was shown by the less sensitive spectrophotometric assay.

Protective Effects of Dietary Garlic Powder Against Cadmium-induced Toxicity in Sea Bass Liver: a Chemical, Biochemical, and Transcriptomic Approach.

To investigate the protective effect of garlic powder on cadmium-induced toxicity sea bass liver, juvenile fishes where maintained under three food diets (diet 1: normal without garlic supply, diet 2: 2% garlic powder; diet 3: 6% garlic powder). After 30 days of specific diets, each group was injected with 500 µg kg-1of Cd. The control group was the one fed with normal diet and not injected with Cd. Liver Cd, Zn, and Se loads was assessed after 1 and 3 days of Cd injections. Moreover, antioxidant enzymes activities termed as catalase, superoxide dismutase, and glutathione peroxydase as well as their gene expression levels were monitored. Finally, metallothionein protein accumulation and its gene expression regulation (MTa) were determined. In fish fed with 2 and 6% garlic powder, the amounts of Cd, Zn, and Se significantly increase in liver tissues. Two percent garlic powder specific diet reversed the Cd-induced inhibition of catalase (CAT), superoxide dismutase (SOD), and gluthathione peroxydase (GPx) and restored the Cd-induced lipid peroxidation (MDA). The increase of liver metallothionein proteins as well as the MTa gene expression level under Cd influence was more pronounced in animals maintained for 30 days under garlic power 2% diet. Our data must be carefully considered in view of the garlic powder introduction in sea bass food composition at 2% since it is an efficient prevention against Cd-induced alterations.

Impact of heavy metal contamination on oxidative stress of Eisenia andrei and bacterial community structure in Tunisian mine soil.

The aims of this work were firstly to study the effect of heavy metal-polluted soils from Tunisian mine on earthworm biochemical biomarkers and on bacterial communities and therefore to analyze the interaction between earth worms and bacterial communities in these contaminated soils. For this purpose, we had introduced earthworm Eisenia andrei in six soils: one from mine spoils and five from agricultural soils, establishing a gradient of contamination. The response of worms to the presence of heavy metal was analyzed at the biochemical and transcriptional levels. In a second time, the impact of worm on bacterial community structure was investigated using automated ribosomal intergenic spacer analysis (ARISA) fingerprinting. An impact of heavy metal-contaminated soils on the oxidative status of E. andrei was observed, but this effect was dependent of the level of heavy metal contamination. Moreover, our results demonstrate that the introduction of earthworms E. andrei has an impact on bacterial community; however, the major change was observed in the less contaminated site. Furthermore, a significant correlation between earthworm oxidative status biomarkers and bacterial community structure was observed, mainly in the mine spoils. Therefore, we contribute to a better understanding of the relationships between epigenic earthworms and bacterial communities in heavy metal-contaminated soils.

Biomarker responses of Eisenia andrei to a polymetallic gradient near a lead mining site in North Tunisia.

Eisenia andrei earthworms were exposed for 7 and 14 days to six samples of soil taken from around an abandoned lead (Pb) mine and characterized by different levels of metal contamination (S6-S1, this latter being the most contaminated soil). The organisms were analyzed for metal bioaccumulation and for biological parameters as biomarkers of stress (lysosomal membrane stability; lipofuscin lysosomal content; lysosomal/cytoplasmic volume ratio) and genotoxicity (Micronucleus frequency). Chemical analysis showed the loads of Pb, Cd, Zn, and Cu in the worms following exposure. Among the stress biomarkers, lysosomal membrane stability was significantly affected in the coelomocytes of the earthworms exposed already 7 days to different contaminated soils. Organisms exposed for 14 days to S1 showed in the cells of the chloragogenous tissue, a particularly relevant increase in lipofuscin, a biomarker of oxidative stress, and an increase in the lysosome/cytoplasm volume ratio, indicating stressful condition at the tissue level. Moreover, in the same conditions, a decrease in total body weight was observed. At the longer exposure time, the coelomocytes of worms exposed to S1, S2, and S3 (soils with higher metal concentrations) showed a significant increase in micronuclei (MNi) frequency. Expressions of the P21 and topoisomerase genes, which are involved in DNA repair, showed significant up-regulation in the cells of worms exposed to S1, S2, S3, S4 and to a less extend S6. This may indicate that the worms were only able to successfully reduce the level of DNA damage in S4 and S5 if considering MN frequency data. The biomarker data was integrated by the Earthworm Expert System, allowing an objective interpretation of the complex biological data and clearly defining the areas in which the presence of chemicals is toxic for the edaphic organisms.

Using environmental proteomics to assess pollutant response of Carcinus maenas along the Tunisian coast.

Biochemical responses to pollutants were studied at four Tunisia littoral sites using Carcinus maenas as a bioindicator. Proteomic analysis was used to assess the global impact of complex pollution mixtures, and to provide new biomarkers and basic insights into pollutant toxicity. Metal contents and metallothionein levels followed a gradient based on sampling sites: Bizerte ≫ Teboulba > Gargour~Mahres. Approximately 900 and 700 spots were resolved in digestive glands and gills, respectively. Gills from Bizerte animals had the maximum number of altered spots, mostly upregulated. In other locations, the number of altered spots in gills decreased in parallel to total metals in in the following order: Teboulba > Gargour > Mahres (mostly downregulated). Out of the 39 spots excised, ten proteins were identified in digestive glands and eight in gills. Digestive glands of Bizerte crabs had higher levels of ferritin, three vitellogenin forms and mannose-binding protein, while Gargour crabs had higher levels of four cryptocyanin forms. Gills of Bizerte crabs had higher levels of ferritin, three vitellogenins forms, lectin 4C, actin, and collagenolytic serine protease. Proteins with altered expression in crabs from Tunisia littoral are related to molting, oxidative stress and inflammation, innate immune response, and proteolysis.

Specific mechanisms of tolerance to copper and cadmium are compromised by a limited concentration of glutathione in alfalfa plants.

The induction of oxidative stress is a characteristic symptom of metal phytotoxicity and is counteracted by antioxidants such as glutathione (GSH) or homoglutathione (hGSH). The depletion of GSH│hGSH in fifteen-day-old alfalfa (Medicago sativa) plants pre-incubated with 1mM buthionine sulfoximine (BSO) affected antioxidant responses in a metal-specific manner under exposure to copper (Cu; 0, 6, 30 and 100µM) or cadmium (Cd; 0, 6 and 30µM) for 7 days. The phytotoxic symptoms observed with excess Cu were accompanied by an inhibition of root glutathione reductase (GR) activity, a response that was augmented in Cd-treated plants but reverted when combined with BSO. The synthesis of phytochelatins (PCs) was induced by Cd, whereas the biothiol concentration decreased in Cu-treated plants, which did not accumulate PCs. The depletion of GSH│hGSH by BSO also produced a strong induction of oxidative stress under excess Cu stress, primarily due to impaired GSH│hGSH-dependent redox homeostasis. In addition, the synthesis of PCs was required for Cd detoxification, apparently also determining the distribution of Cd in plants, as less metal was translocated to the shoots in BSO-incubated plants. Therefore, specific GSH│hGSH-associated mechanisms of tolerance were triggered by stress due to each metal.

Comparative study of the bioaccumulation and elimination of trace metals (Cd, Pb, Zn, Mn and Fe) in the digestive gland, gills and muscle of bivalve Pinna nobilis during a field transplant experiment.

The purpose of this study was to investigate the long-term bioaccumulation and elimination of Cd, Pb, Mn, Zn and Fe by Pinna nobilis tissues after their 90 day-transplantation period at Téboulba fishing harbor. During the transplantation period, the Cd, Pb, Mn, Zn and Fe concentrations in the different tissues of the mussels were measured before and after exposure period. Metal (Cd, Pb, Mn, Zn and Fe) accumulation in P. nobilis mussels varied depending on the analyzed tissue and the caging times. Notable differences in Cd, Pb, Mn, Zn and Fe accumulation patterns within the digestive gland, gills and muscle were found and may be due to the ability of each tissue to accumulate metals. During the depuration phase, the elimination of Cd, Pb, Mn, Zn and Fe depended on the target tissue and the metal speciation. Cd, Pb, Mn and Fe were eliminated rapidly from one organ and increased in other when compared to those of 90 day transplanted mussels. The increase of metal loads during the elimination phase is not clear and particularly what kind of processes is responsible for such response. However, it is reasonable to assume that metals increase is related to the existence of an accumulation/detoxification mechanism, which involves the transport of metals from an organ to another. The data obtained indicate that because of the significantly high quantities of Cd, Pb, Mn, Zn and Fe accumulated during the exposure phase, the transplanted mussels are suitable bioindicators for monitoring trace metals in marine ecosystem.