Isolation and characterisation of metallothionein from the clam Ruditapes decussatus

Metallothioneins (MT) were obtained after purification from metal-exposed clams (Ruditapes decussatus) using gel-permeation and ion-exchange chromatography. Four cadmium-metallothioneins (CdMTs) were resolved by ion-exchange chromatography and they all had similar molecular weights, high cadmium content and an absorption spectra indicative of the presence of characteristic Cd-S aggregates. The NH2-terminal sequence suggests the presence of at least two class I clam MT isoforms. For the other two putative clam CdMTs isolated, the results of the amino acid determination were inconclusive. One was slightly contaminated and the other one had a blocked NH2-terminal. These clam metalothioneins contain glycine, which seems to be a common feature of molluscan MT family and exhibited more similarity to oysters than to mussels. Further investigation on the inducibility of these isoforms will be necessary if clams are to be used as biomarkers of metal exposure

Accumulation of Heavy Metals in Franciscanas from Buenos Aires Province, Argentina

Marine mammals accumulate heavy metals in their tissues at different concentrations according to trophic levels and environmental conditions. The franciscana (Pontoporia blainvillei) is a small coastal species inhabiting the marine and estuarine areas of the Southwestern Atlantic Ocean. Its diet includes numerous species of small fish, squid and crustaceans. The aims of this study were to (i) assess the heavy metal concentration and burden distribution in different franciscana age classes and sex, and to (ii) evaluate both the accumulation processes and the transplacental transference of zinc, cadmium, copper and total mercury. Heavy metal concentrations (wet weight) were determined in eighteen dolphins by Atomic Absorption Spectrophotometry (AAS), by the cold vapour technique (mercury) or with air/acetylene flame (cadmium, zinc and copper). Liver showed the highest concentrations of mercury (max. 8.8 mg/g), zinc (max. 29.7 mg/g) and copper (max. 19.0 mg/g), whereas the highest cadmium concentrations (max. 6.7 mg/g) were found in kidney. Adults contained the highest concentrations for all heavy metals, followed by juveniles and calves in decreasing order, suggesting an age-related accumulation. No differences (p<0.05) were found between sexes within each age class. Organ burden distribution followed the same pattern for all metals and age classes: liver tissues contained maximum burdens. Mercury concentrations were higher than those of cadmium in both foetuses and newborns; and neither metal could be detected in the foetus. The analysed data suggested differences in the placental transference between metals, being significant for mercury and almost null in the case of cadmium. We can conclude that franciscana accumulates heavy metals and, due to its coastal distribution, it may be considered as a biomonitor of its environment. SPANISH: Los mamíferos marinos acumulan metales pesados en sus tejidos cuyas concentraciones están en relación con su nivel trófico y las condiciones ambientales. La franciscana (Pontoporia blainvillei) es una especie costera que habita áreas marinas y estuariales en el Atlántico Sudoccidental. Su dieta está constituída por peces, como item alimentario principal, calamares y crustáceos. El objetivo del presente trabajo es estudiar la distribución de metales pesados en diferentes clases de edad y en ambos sexos, evaluando procesos de acumulación y cargas de cadmio, mercurio total, cinc y cobre. Las concentraciones de metales pesados (en peso húmedo) fueron determinadas en dieciocho delfines por Espectrofotometría de Absorción Atómica (EAA), usando la técnica de vapor frío (mercurio) o llama de aire/acetileno (cadmio, cinc y cobre). El hígado presentó las concentraciones más altas de mercurio (máx. 8,8 mg/g), cinc (máx. 29,7 mg/g) y cobre (máx. 19,0 mg/g), mientras que las más altas de cadmio (máx. 6,7 mg/g) fueron encontradas en el riñón. Los adultos presentaron los niveles más altos, presentando los juveniles y cachorros concentraciones menores, lo cual sugirió una acumulación con la edad. No se encontraron diferencias significativas (p < 0,05) entre sexos dentro de cada clase de edad. Las cargas de metales pesados en los órganos presentaron la misma disribución para todos los metales y clases de edad. Los valores más altos fueron encontrados en el hígado, incluyendo los correspondientes a cadmio. Las concentraciones de mercurio y cadmio fueron no detectables en el feto, mientras que las de mercurio fueron superiores a las de cadmio en los cachorros. Los datos encontrados en el feto sugieren una transferencia nula a través de la placenta. Podemos concluir que P.blainvillei acumula metales pesados en sus tejidos y debido a su distribución costera, esta especie puede ser considerada como un biomonitor de su ambiente

Genotoxic and stress inductive potential of cadmium in Xenopus laevis larvae

The present investigation evaluates the toxic potential of Cd in larvae of the frog Xenopus laevis after 12 days of exposure to environmentally relevant contamination levels, close to those measured in the river Lot (France). Several genotoxic and detoxification mechanisms were analyzed in the larvae: clastogenic and/or aneugenic effects in the circulating blood by micronucleus (MN) induction, metallothionein (MT) production in whole larvae, gene analyses and Cd content in the liver and also in the whole larvae. The results show: (i) micronucleus induction at environmental levels of Cd contamination (2, 10, 30 μg L−1); (ii) an increased and concentration-dependent quantity of MT in the whole organism after contamination with 10 and 30 μg Cd L−1 (a three- and six-fold increase, respectively) although no significant difference was observed after contamination with 2 μg Cd L−1; (iii) Cd uptake by the whole organism and by the liver as a response to Cd exposure conditions; (4) up-regulation of the genes involved in detoxification processes and response to oxidative stress, while genes involved in DNA repair and apoptosis were repressed. The results confirm the relevance of the amphibian model and highlight the complementarity between a marker of genotoxicity, MT production, bioaccumulation and genetic analysis in the evaluation of the ecotoxicological impact

Biomarkers: a strategic tool in the assessment of environmental quality of coastal waters

Ecosystems are under the pressure of complex mixtures of contaminants whose effects are not always simple to assess. Biomarkers, acting as early warning signals of the presence of potentially toxic xenobiotics, are useful tools for assessing either exposure to, or the effects of these compounds providing information about the toxicant bioavailability. In fact, it has been argued that a full understanding of ecotoxicological processes must consider an integrated multi-level approach, in which molecular impact is related with higher-order biological consequences at the individual, population and community levels. Monitoring programs should make use of this tool to link contaminants and ecological responses fulfilling strategies like those launched by OSPAR (Commissions of Oslo and Paris) Convention on the protection of the marine environment of the North-East Atlantic and the International Council for the Exploration of the Sea (ICES). An overview of the work done in the past few years using biomarkers as in situ tools for pollution assessment in Portuguese coastal waters is presented as a contribution to the set up of a biomonitoring program for the Portuguese coastal zone. Considering the data set available the biomonitoring proposal should include the analysis of biomarkers and effects at individual levels. The aim of the program will include a spatial and temporal characterization of the biomarkers acetyl-cholinesterase, metallothioneins, DNA damage, adenylate energy charge and scope-for-growth levels. The investigation of the spatial variation of biomarkers is crucial to define sites for long term monitoring, which will be integrated with a chemical monitoring program. This framework will be a major contribution to the implementation of a national database for the use of biomarkers along the Portuguese coast.info:eu-repo/semantics/publishedVersio

Heavy metal intoxication

Although some debate exists with regards to the subject, elements which are classified under ‘heavy metals’ have come to be those which pose a threat to humans in terms of toxicity. Intoxication with heavy metals is not a typical diagnosis as it is fairly uncommon. This can impose a risk on people who fail to be diagnosed and removed from the source of exposure, increasing morbidity and mortality. For the purposes of this review, Cadmium and Mercury will be discussed. A brief introduction of each element’s chemical and physical properties will be given, as well as its sources in the environment and any uses. Each metal’s toxicity will be illustrated using actual cases of poisoning. Any treatments for intoxication will be explained at the end of each section.peer-reviewe

Metallothionein as an indicator of water quality: assessment of the bioavailability of cadmium, copper, mercury and zinc in aquatic animals at the cellular level

The study of metallothioneins (MTs) has greatly improved our understanding of body burdens, metal storage and detoxification in aquatic organisms subjected to contamination by the toxic heavy metals, Cd, Cu, Hg and Zn. These studies have shown that in certain organisms MT status can be used to assess impact of these metals at the cellular level and, whilst validation is currently limited to a few examples, this stress response may be linked to higher levels of organisation, thus indicating its potential for environmental quality assessment. Molluscs, such as Mytilus spp., and several commonly occurring teleost species, are the most promising of the indicator species tested. Natural variability of MT levels caused by the organism's size, condition, age, position in the sexual cycle, temperature and various stressors, can lead to difficulties in interpretation of field data as a definitive response-indicator of metal contamination unless a critical appraisal of these variables is available. From laboratory and field studies these data are almost complete for teleost fish. Whilst for molluscs much of this information is lacking, when suitable controls are utilised and MT measurements are combined with observations of metal partitioning, current studies indicate that they are nevertheless a powerful tool in the interpretation of impact, and may prove useful in water quality assessment

Long-chain class III metallothioneins as a mechanism of cadmium tolerance in the marine diatom Phaeodactylum tricornutum Bohlin

Cadmium tolerance of the marine diatom Phaeodactylum tricornutum Bohlin was studied. P. tricornutum growth was significantly reduced by cadmium concentrations of 5 mg l−1 or higher. The EC50 value was 22.39 mg l−1 of cadmium after four days of exposure. Therefore, P. tricornutum is more tolerant to cadmium than other microalgal cells. In response to exposure to cadmium, P. tricornutum synthesize class III metallothioneins which sequester the metal in a harmless form. However, cadmium tolerance of P. tricornutum cells is not only due to the ability of this microalga to synthesize class III metallothioneins (γ-Glu-Cys)n-Gly, but also because these metallothioneins are composed of long-chain polypeptides. The cadmium-metallothionein complexes of P. tricornutum cells contain predominantly polypeptides of n = 4−9. Long-chain metallothioneins are more efficient in binding heavy metals. Capillary electrophoresis was used to separate class III metallothioneins. This is a recent technique used to separate many molecules. With this technique, the length of the separated polypeptides can also be estimated.Xunta de Galicia; XUGA 10301B9

Metallothioneins in the clam Ruditapes decussatus: an overview

The clam Ruditapes decussatus is a suspension-feeding bivalve mollusc widely distributed in European waters and in the Mediterranean. Due to is economic importance it is heavily harvested in many countries, and particularly in Portugal. Its ability to accumulate high metal concentrations along with its economic importance was the main reason for its selection as a bioindicator. Metallothionein (MT) concentrations in the clams R. decussatus followed by gel filtration chromatography, differential pulse polarography and SDS-PAGE, after Cd exposure, revealed that MT is induced in different tissues (whole soft tissues, gills, digestive gland and remaining tissues) but the level of MT induction is tissue dependent. MT from the gills and the digestive gland give a more sensitive response to assess the effects of metal exposure directly from the water or from the food than in the whole soft tissues. MT levels were also measured in the gills, digestive gland and remaining tissues of R. decussatus collected in the Ria Formosa lagoon (Portugal) from areas of different metal load and during the period of sexual differentiation of the clam. Data revealed that there were significant differences of MT concentrations among sites and season but not among sex. Purification of MT from the digestive gland of R. decussatus revealed four MT isoforms. The molecular weight of one of these isoforms, determined by SDS-PAGE, was of the same order of magnitude as that of MT from other bivalve species. Similarly the amino acid sequence of the beta domain of the MT of the digestive gland of the clam also shows some degree of similarity with the similar MT sequence from mussels and oysters. It is, therefore suggested that there is some degree of similarity in the MT structure among these species.info:eu-repo/semantics/publishedVersio