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- A multibiomarker approach in the clam Ruditapes decussatus to assess the impact of pollution in the Ria Formosa lagoon, South Coast of PortugalPublication . Cravo, Alexandra; Pereira, C.; Gomes, Tânia; Cardoso, Cátia; Serafim, M.A.; Almeida, Cheila; Rocha, T.; Lopes, Belisandra; Company, Rui; Medeiros, A.; Norberto, R.; Pereira, R.; Araújo, O.; Bebianno, Maria JoãoThe Ria Formosa lagoon is an ecosystem whose water quality reflects the anthropogenic influence upon the surrounding areas. In this lagoon, the clam Ruditapes decussatus has a great economical importance and has been widely used as a biomonitor. A multibiomarker approach (d-aminolevulinic acid dehy- dratase, metallothionein, lipid peroxidation, acetylcholinesterase, alkali-labile phosphates, DNA damage) was applied to assess the environmental quality of this ecosystem and the accumulation of contaminants and their potential adverse effects on clams. Clams were sampled in different shellfish beds in the period between July 2007 and December 2008 and abiotic parameters (temperature, salinity, pH and dissolved oxygen of seawater and organic matter in the sediment), condition index, metals (Cd, Cu, Zn, Ni, Pb), TBTs and PAHs concentrations were measured in clam tissues. Data was integrated using Principal Component Analyses and biomarker indices: IBR (Integrated Biomarker Response) and HSI (Health Status Index). This multibiomarker approach enabled discrimination of a time and space trend between sites with different degrees of anthropogenic contamination, identifying one of them (site 2) as the most stressful and summer months as the most critical period for clams due to an increase of environmental stress (anthropogenic pressure along with extreme environmental conditions, e.g. temperature, dissolved oxygen, organic matter in the sediments, etc). The selected biomarkers provided an integrated response to assess the environmental quality of the system, proving to be a useful approach when complex mixtures of contaminants occur.
- Comparison of metallothionein induction in response to cadmium in the gills of the bivalve molluscs Mytilus galloprovincialis and Ruditapes decussatusPublication . Bebianno, Maria João; Serafim, A.Metal binding characteristics and metallothionein induction differ markedly among the gills of the bivalve molluscs Mytilus galloprovincialis and Ruditapes decussatus exposed to sublethal cadmium concentrations (100 μg/l). The rates of cadmium uptake as well as the percentage of cadmium bound to the cytosol were significantly higher in the gills of M. galloprovincialis when compared with that of R. decussatus. Similarly metallothionein concentration detected in the gills of M. galloprovincialis increase fourfold while for the clam R. decussatus metallothionein concentrations increased only twofold.
- Effect of a polymetallic mixture on metal accumulation and metallothionein response in the clam Ruditapes decussatusPublication . Serafim, Angela; Bebianno, Maria JoãoThe toxic effects of metals are related to changes in natural physiological and biochemical processes in organisms, which are not easily detected. Marine invertebrates developed strategies to survive in the presence of toxic metal levels, by the removal of these compounds from the soluble fraction of the cell through different processes, including the synthesis of metallothionein (MT). To better understand the effect of Cd, Cu and Zn in MT response it is important to consider that in their natural environment these metals are normally mixed. The exposure to a metal mixture result in toxicological interactions that produce different biological responses when compared to the effects of a single metal exposure. The aim of this study was to assess metal accumulation and MT response in different tissues of the clam Ruditapes decussatus exposed to a mixture of sublethal Cd, Cu and Zn concentrations. Kinetic models were applied to determine metal uptake and loss to better predict and understand the detoxification mechanisms in this species. The kinetic model showed that the effect of this polymetallic exposure compared to a single exposure to these metals caused different metal uptake and loss rates in several tissues. Moreover, the exposure of R. decussatus to a polymetallic mixture caused an increase in MT induction compared to a single metal exposure. This is probably related to the interaction between the different metals and their different affinity to this protein. Metal interactions within aquatic organisms are very important in MT synthesis and the effect of polymetallic mixtures in the environment must be taken into account in field studies.
- Metalotionina como biomarcador de contaminação metálica na amêijoa Ruditapes decussatusPublication . Serafim, M.A.; Bebianno, Maria JoãoO objectivo da presente dissertação foi contribuir para a compreensão da aplicação da metalotionina (MT) como biomarcador de contaminação metálica na amêijoa Ruditapes decussatus.
- Biomarkers in Ruditapes decussatus: a potential bioindicator speciesPublication . Bebianno, Maria João; Géret, F.; Hoarau, P.; Serafim, Angela; Coelho, M. R.; Gnassia-barelli, M.; Roméo, M.The clam Ruditapes decussatus is distributed worldwide and due to its ecological and economical interest has been proposed as a bioindicator in areas where mussels are not available. The accumulation of several anthropogenic compounds in their tissues suggests that they possess mechanisms that allow them to cope with the toxic effects of these contaminants. Besides pollutant uptake, the use of biomarkers is pointed out in this paper since it is a promising approach to monitor the effect of these contaminants in the marine environment. Biomarkers complement the information of the direct chemical characterization of different types of contaminants. Therefore, the aim of this paper is to review the role of several biomarkers: (metallothioneins (MT), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx) (total and selenium-dependent), lipid peroxidation (measured as MDA, one of the final products of lipid peroxidation), glutathione S-transferase (GST) and acetylcholinesterase (AChE), measured in different tissues of the clam R. decussatus, in laboratory conditions and under various environmental stresses, in two ecosystems (Ria Formosa lagoon- Portugal) and Bizerta lagoon (Tunisia) in a perspective of a multibiomarker approach to assess environmental changes. Experiment and field studies are in good agreement since MT levels, especially in the gills, the first target tissue of these contaminants, can be used as biomarker of exposure to Cd. GPx and MDA may also be determined in this respect. AChE activity is inhibited by pesticide and, to a less extent, by metal exposure in the gills and whole soft body of clams. However, the induction of GST isoforms experimentally demonstrated is not observed in the field because only global GST activity was determined. The whole set of results opens new research perspectives for the use of this species to assess the effect of mixtures of pollutants in the aquatic environment.
- Variation of metallothionein and metal concentrations in the digestive gland of the clamRuditapes decussatus: sex and seasonal effectsPublication . Serafim, Angela; Bebianno, Maria JoãoMetallothionein (MT) and metal (Cd, Cu, and Zn) concentrations were determined in the digestive gland of male and female Ruditapes decussates. Clams were collected monthly during the period of sexual development, from June to September, at two different sites of a coastal lagoon, the Ria Formosa Lagoon, on the south coast of Portugal. The MT concentrations were determined in the heat-treated cytosolic fraction of the digestive gland of both male and female clams. Total metal (Cd, Cu, and Zn) concentrations in the heat-treated cytosol of the digestive gland were also determined in both sexes. The MT and metal concentrations in the digestive gland were not sex dependent in this species. Therefore, these results suggest that random samples can be used to determine MT and metal concentrations in the digestive gland of R. decussatus. Seasonal and site-specific dependency, however, were detected for MT, Cd, and Cu concentrations. The maximum concentration levels of MT, Cd, and Cu were detected in July and August for both sexes in the same area. Zinc concentrations, however, did not show marked seasonal variations, indicating that this species is able to regulate zinc concentrations. The highest MT concentrations were significantly related to the highest metal concentrations.
- Kinetic model of cadmium accumulation and elimination and metallothionein response in Ruditapes decussatusPublication . Serafim, Angela; Bebianno, Maria JoãoThe aim of the present study was to determine the response of metallothionein (MT) during Cd accumulation and elimination in different tissues of the estuarine bivalve Ruditapes decussatus exposed to two nominal Cd concentrations (4 and 40 microg/L) for 40 d, followed by a depuration period of 50 d. Cadmium was accumulated in all tissues of R. decussatus at both exposure concentrations, and the accumulation was tissue dependent. Use of the kinetic model showed that in the gills and remaining tissues, Cd was assimilated faster at the beginning of the exposure and decreased with time, possibly limited by the diffusion rate of this metal within the cell. In the digestive gland, however, the Cd was continuously accumulated. This could reflect that the Cd uptake rate is considerably higher than the loss rate and, therefore, that this tissue has a higher capacity to accumulate Cd compared to the other two tissues. Moreover, the application of this kinetic model in the different subcellular fractions showed that the bioconcentration factor was significantly higher in the low-molecular-weight fraction (where MT is found), suggesting that this fraction binds Cd faster, with a high uptake rate (K(u) = 32/d), and eliminates this metal more slowly (K(1) = 0.005/d). During the depuration phase, MT decreased simultaneously with Cd elimination in all tissues, although with a shorter half-life. In conclusion, the MT response prevented Cd in the tissues of R. decussatus from interfering in the normal clam metabolism; therefore, MT acts as a detoxification mechanism of Cd.