Analysis of proteins from marine molluscs

Application of the two-dimensional gel electrophoresis (2DE) protocols which were developed for samples of mammalian origin gives unsatisfactory results when used in samples from marine molluscs. This chapter describes a detailed protocol of 2DE that can be applied to these organisms, especially for Ruditapes decussatus and Bathymodiolus azoricus.info:eu-repo/semantics/publishedVersio

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

Partitioning of trace metals between soft tissues and shells of Patella aspera

This paper describes the partitioning of iron, manganese, zinc, copper, cadmium, cobalt, nickel and lead between the soft tissues and the shells of the gastropod mollusc Patella aspera. Specimens of the limpet P aspera were collected from a clean coastal marine site (CMS) and from an estuarine contaminated site (ECS) on the south coast of Portugal. Fe and Zn concentrations were significantly lower on both populations and less variable in the shells than in the soft tissues. In contrast Mn concentrations were higher in the shells than in the soft tissues at ECS. The partitioning factor (PF), defined as the ratio between the mean metal concentrations in soft tissues and in the shells, was maximum for iron and minimum for manganese. Between the soft tissues and the shells, only manganese at ECS recorded a significant relationship between metal concentrations, suggesting that the mechanism that controls the accumulation of this essential metal in both tissues was different from the others. From the obtained data, in biomonitoring sewage contamination studies, shells of P. aspera can be considered as a good indicator for Mn while the soft tissues for Zn. As copper, cadmium, nickel and cobalt concentrations were much higher in the soft tissues than in the shells ( < 0.1 mug g(-1)), this indicates that the shells were not a good indicator for these metals because they do not reflect the environmental bioavailability as do the soft tissues.FRMH/BD/403/92info:eu-repo/semantics/publishedVersio

Responses of CYP450 dependent system to aliphatic and aromatic hydrocarbons body burden in transplanted mussels from South coast of Portugal

Mussels Mytilus galloprovincialis were cross-transplanted at South Portugal from a reference site (site 1) to a site more contaminated with hydrocarbon compounds (site 2), and vice versa, in an active biomonitoring (ABM) concept, to assess the biotransformation capacity catalyzed by the mixed function oxygenase (MFO) system. Total alkanes (TAlk), the unresolved complex mixture (UCM), and total polycyclic aromatic hydrocarbons (TPAHs) concentration increased respectively 6, 4.4 and 4.2 fold relatively to control, in mussels transplanted from site 1 to 2. In the cross-transplant, a 48, 57 and 62% depuration of TAlk, UCM and TPAHs concentrations occurred by the end of the 3-4th week. Petrogenic and biogenic (marine and terrigenous) sources of AHs, and petrogenic and pyrolitic (biomass and oil/fuel incomplete combustion) sources of PAHs were detected at both sites. CYP450, CYT b (5) and NADPH-RED in mussels transplanted from site 1 to 2 were induced from day 0 to 28, with a total increase of 35, 32 and 35%, respectively, while biochemical equilibrium to lesser environmental contamination occurs in mussels transplanted from site 2 to 1. A significant relationship between CYP450 and NADPH-RED was found with TPAH, with distinctive behavior at the two sites. MFO system components increase with exposure time at one site and decreases in the other, reflecting an adaptation to distinct environmental hydrocarbon loads. The ABM strategy proved to be useful to understand the environment real impact on the biochemical responses in mussels' local populations. In this study, CYP450 and NADPH-RED are a useful biomarker for hydrocarbon exposure.FCT: PRAXIS XXI/BD/3740/94info:eu-repo/semantics/publishedVersio

Problemas de contaminação na zona costeira : os desafios do século XXI

No artigo são enunciados alguns dos desafios que o século XXI apresenta em termos da poluição dos oceanos, muito em particular da zona costeira. São referidas algumas fontes de contaminação das águas, que constituem outras tantas ameaças: lançamento ao mar de metais pesados, acidentes com petroleiros, acidentes em centrais nucleares. São igualmente mencionados os efeitos de grandes perturbações ambientais, o desenvolvimento industrial e correspondente aumento populacional nas zonas costeiras. O homem é o responsável pela situação de poluição com mercúrio, cobre, cádmio e essa substância terrível que é o TBT, apresentando exemplos como o da Ria Formosa e outros locais da costa algarvia onde se verificam níveis superiores aos normais. Por fim refere algumas medidas para a eliminação das substâncias nocivas do ambiente aquático, apontando a necessidade de coordenação dos vários organismos responsáveis ou mesmo da sua fusão para conseguir uma maior eficácia de actuação

The composition of phosphate granules in the digestive glands of marine prosobranch gastropods: variation in relation to taxonomy

The composition of some 1150 phosphate granules in the digestive glands of over 40 species of marine prosobranch gastropods has been surveyed using a simple preparation technique and semiquantitative SEM x-ray microanalysis. Spectral peaks for Mg, K, Ca, Mn, Fe and Zn were compared to that of P. Four major types of phosphate granule can be recognised, each generally characteristic of a taxonomic grouping: high Mg in archaeogastropods and littorinids, multiple metal in higher mesogastropods, and, in neogastropods, Mg-Ca in muricoideans and high Zn in buccinoideans. At least one Conus species (C. ventricosus) has high-Mg granules. Some causes of variation in granule composition are discussed: speculatively, it is suggested a palaeoenvironmental influence seems possible.info:eu-repo/semantics/publishedVersio

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

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

Transcriptomic effects of the non-steroidal anti-inflammatory drug Ibuprofen in the marine bivalve Mytilus galloprovincialis Lam

The transcriptomic effects of Ibuprofen (IBU) in the digestive gland tissue of Mytilus galloprovincialis Lam. specimens exposed at low environmental concentrations (250 ng L-1) are presented. Using a 1.7 K feature cDNA microarray along with linear models and empirical Bayes statistical methods 225 differentially expressed genes were identified in mussels treated with IBU across a 15-day period. Transcriptional dynamics were typical of an adaptive response with a peak of gene expression change at day 7 (177 features, representing about 11% of sequences available for analysis) and an almost full recovery at the end of the exposure period. Functional genomics by means of Gene Ontology term analysis unraveled typical mussel stress responses i.e. aminoglycan (chitin) metabolic processes but also more specific effects such as the regulation of NF-kappa B transcription factor activity. (C) 2016 Elsevier Ltd. All rights reserved