Alzheimer's Disease and Toxins Produced by Marine Dinoflagellates: An Issue to Explore.

This paper examined the toxins naturally produced by marine dinoflagellates and their effects on increases in ß-amyloid plaques along with tau protein hyperphosphorylation, both major drivers of Alzheimer's disease (AD). This approach is in line with the demand for certain natural compounds, namely those produced by marine invertebrates that have the potential to be used in the treatment of AD. Current advances in AD treatment are discussed as well as the main factors that potentially affect the puzzling global AD pattern. This study focused on yessotoxins (YTXs), gymnodimine (GYM), spirolides (SPXs), and gambierol, all toxins that have been shown to reduce ß-amyloid plaques and tau hyperphosphorylation, thus preventing the neuronal or synaptic dysfunction that ultimately causes the cell death associated with AD (or other neurodegenerative diseases). Another group of toxins described, okadaic acid (OA) and its derivatives, inhibit protein phosphatase activity, which facilitates the presence of phosphorylated tau proteins. A few studies have used OA to trigger AD in zebrafish, providing an opportunity to test in vivo the effectiveness of new drugs in treating or attenuating AD. Constraints on the production of marine toxins for use in these tests have been considered. Different lines of research are anticipated regarding the action of the two groups of toxins.

Factors influencing sorption of trace elements in contaminated waters onto ground nut shells.

Biosorbent materials such as nut shells has been considered a promising alternative to the classic methods to remove potentially toxic elements from contaminated waters, improving water quality for other uses. The present study evaluated the sorption capacity of almond, hazelnut, pistachio and walnut shells for the single and simultaneous removal of As, Cd, Co, Hg, Ni, Pb and Zn from contaminated waters. The influence of key parameters such as sorbent dose, ionic strength (ultrapure water vs. mineral water), element competition (mono-vs. multi-element spiked solution) and initial element concentration (maximum allowed value in wastewater discharges and ten times lower) was assessed. Hazelnut shells stood out as the material with higher potential to remove Cd, Co, Hg, Ni, Pb and Zn in the tested conditions, achieving up to 99% of removal (Cd and Pb). Arsenic was not removed by none of the studied materials and the removal of Cr was only obtained in a simple matrix and in mono-element solution (41% of removal). An increase of matrix complexity negatively affected the sorption capacity of all the biosorbents in the removal of all the elements tested. The same behavior was obtained when the elements were simultaneously present in solution, with Pb, Cd and Hg being highly influenced by the presence of other elements. The increase in sorbent dose from 1 to 5 g/L allowed to at least duplicate the removal of Cd, Co, Ni and Zn by all the biosorbents. When decreasing the initial element concentration ten times the maximum allowed value in wastewater discharges, a significant increase of elements removal was attained, more pronounced for Hg and Pb. The possibility to change initial conditions towards high efficiencies validate the high potential of those biosorbents to be used in water remediation, with special focus on hazelnut shells, although other sorbents such as walnut or almond shells can be considered efficient with appropriate initial conditions and depending on the target elements to be removed.

Major characteristics of microplastics in mussels from the Portuguese coast.

The present study reports the quantity, shape, colour and chemical properties of microplastics (MP) and MP-like in whole soft tissues of the mussel Mytilus spp. collected in January and February 2019 from four natural banks in the Portuguese coast. Three sites are located in estuarine areas influenced by anthropogenic pressures and freshwater discharges, and one in the coast far from urbanised areas. An alkaline digestion (KOH) of biological tissues was used and a polymeric identification of 20% of the visually sorted particles was achieved using the Fourier-transform mid-infrared spectroscopy (FT-MIR). MP and MP-like concentrations ranged from 0.54 to 3.0 items g-1 without significant differences among the sites. Particle size varied from 36 to 4439 µm, being fibers the most abundant shape (50%) followed by films (22%) and spherules (18%). FT-MIR revealed that 69% of the analysed particles were plastic, being identified six polymers and two polymeric blends, and 32% were cellulose-based materials. Fibers identified in mussel tissues were mainly composed of cotton and viscose (77%). This study emphasizes the importance of the polymer's spectroscopic identification after microscopic observation to recognise MP.

Biochemical performance of mussels, cockles and razor shells contaminated by paralytic shellfish toxins.

Marine toxins in bivalves pose an important risk to human health, and regulatory authorities throughout the world impose maximum toxicity values. In general, bivalve toxicities due to paralytic shellfish toxins (PSTs) above the regulatory limit occur during short periods, but in some cases, it may be extended from weeks to months. The present study examines whether cockles (Cerastoderme edule), mussels (Mytilus galloprovincialis) and razor shells (Solen marginatus) naturally exposed to a bloom of Gymnodinium catenatum activated or suppressed biochemical responses as result of the presence of PSTs in their soft tissues. Toxins (C1+2, C3+4, GTX5, GTX6, dcSTX, dcGTX2+3 and dcNEO) and a set of biomarkers (ETS, electron transport system activity; GLY, glycogen; PROT, protein; SOD, superoxide dismutase; CAT, catalase; GPx, glutathione peroxidase; GST, glutathione S-transferases; LPO, lipid peroxidation; reduced (GSH) and oxidized (GSSG) glutathione contents and AChE, acetylcholinesterase activity) were determined in the three bivalve species. Specimens were harvested weekly in Aveiro lagoon, Portugal, along thirteen weeks. This period included three weeks in which bivalve toxicity exceeded largely the regulatory limit and the subsequence recovery period of ten weeks. Biochemical performance of the surveyed species clearly indicated that PSTs induce oxidative stress and neurotoxicity, with higher impact on mussels and razor shells than in cockles. The antioxidant enzymes CAT and GPx seemed to be the biomarkers better associated with toxin effects.

A green method based on living macroalgae for the removal of rare-earth elements from contaminated waters.

Low recycling rates of rare earth elements (REEs) are a consequence of inefficient, expensive and/or contaminating methods currently available for their extraction from solid wastes or from liquid wastes such as acid mine drainage or industrial wastewaters. The search for sustainable recovery alternatives was the motivation for this study. For the first time, the capabilities of 6 living macroalgae (Ulva lactuca, Ulva intestinalis, Fucus spiralis, Fucus vesiculosus, Osmundea pinnatifida and Gracilaria sp.) to remove REEs (Y, La, Ce, Pr, Nd, Eu, Gd, Tb, Dy) from laboratory-prepared seawater spiked with REE solutions were evaluated. The assays lasted 72 h with REEs concentrations ranging from 10 to 500 µg L-1. The link between REEs uptake and algal metabolism, surface morphology and chemistry were addressed. Kinetics varied among the species, although most of the removal occurred in the first 24 h, with no equilibrium being reached. Lack of mortality reveal that the algae maintained their metabolism in the presence of the REEs. Green alga U. lactuca stood out as the only capable of efficiently removing at least 60% of all elements, reaching removals up to 90% in some cases. The high bioconcentration factors, derived from mass balance analysis (c.a. 2500) support that the REEs enriched algal biomass (up to 1295 µg g-1) may constitute an effective and environmentally friendly alternative source of REEs to conventional extraction from ores.

Experimental Measurement and Modeling of Hg(II) Removal from Aqueous Solutions Using Eucalyptus globulus Bark: Effect of pH, Salinity and Biosorbent Dosage.

Different experimental conditions were tested in order to optimize the Hg(II) removal by Eucalyptus globulus bark. Response surface methodology was applied to extract information about the significance of the factors and to obtain a model describing the sorption. The results were generated through the design of experiments by applying the methodology of a three-factor and three-level Box-Behnken design. The factors tested were pH (4.0, 6.5, and 9.0), salinity (0, 15, and 30), and biosorbent dosage (0.2, 0.5, and 0.8 g dm-3) to evaluate the Hg(II) removal using realistic conditions, such as contaminated natural waters with an initial Hg(II) concentration of 50 µg dm-3. The optimum response provided by the model was 81% of the metal removal under the optimal operating conditions: a pH value of 6.0, no salinity, and a biosorbent dosage of 0.55 g dm-3. Concerning the kinetic, the pseudo-second-order equation fitted better to the experimental results with R 2 between 0.973 and 0.996. This work highlights the promising valorization of this biomass, which is an industrial byproduct and makes available information about the influence of the variables for Hg(II) removal in water treatment processes.

A macroalgae-based biotechnology for water remediation: Simultaneous removal of Cd, Pb and Hg by living Ulva lactuca.

Metal uptake from contaminated waters by living Ulva lactuca was studied during 6 days, under different relevant contamination scenarios. In mono-metallic solutions, with concentrations ranging from 10 to 100 µg L-1 for Hg, 10-200 µg L-1 for Cd, and 50-1000 µg L-1 for Pb, macroalgae (500 mg L-1, d.w.) were able to remove, in most cases 93-99% of metal, allowing to achieve water quality criteria regarding both surface and drinking waters. In multi-metallic solutions, comprising simultaneously the three metals, living macroalgae still performed well, with Hg removal (c.a. 99%) not being significantly affected by the presence of Cd and Pb, even when those metals were in higher concentrations. Removal efficiencies for Cd and Pb varied between 57 and 96%, and 34-97%, respectively, revealing an affinity of U. lactuca toward metals: Hg > Cd > Pb. Chemical quantification in macroalgae, after bioaccumulation assays demonstrated that all Cd and Hg removed from solution was really bound in macroalgae biomass, while only half of Pb showed to be sorbed on the biomass. Overall, U. lactuca accumulated up to 209 µg g-1 of Hg, up to 347 µg g-1 of Cd and up to 1641 µg g-1 of Pb, which correspond to bioconcentration factors ranging from 500 to 2200, in a dose-dependent accumulation. Pseudo-first order, pseudo-second order and Elovich models showed a good performance in describing the kinetics of bioaccumulation, in the whole period of time. In the range of experimental conditions used, no mortality was observed and U. lactuca relative growth rate was not significantly affected by the presence of metals. Results represent an important contribution for developing a macroalgae-based biotechnology, applied for contaminated saline water remediation, more "green" and cost-effective than conventional treatment methods.

Superoxide dismutase enzymosomes: carrier capacity optimization, in vivo behaviour and therapeutic activity.

PURPOSE: A strategy not usually used to improve carrier-mediated delivery of therapeutic enzymes is the attachment of the enzymes to the outer surface of liposomes. The aim of our work was to design a new type of enzymosomes with a sufficient surface-exposed enzyme load while preserving the structural integrity of the liposomal particles and activity of the enzyme. METHODS: The therapeutic antioxidant enzyme superoxide dismutase (SOD) was covalently attached to the distal terminus of polyethylene glycol (PEG) polymer chains, located at the surface of lipid vesicles, to obtain SOD-enzymosomes. RESULTS: The in vivo fate of the optimized SOD-enzymosomes showed that SOD attachment at the end of the activated PEG slightly reduced the residence time of the liposome particles in the bloodstream after IV administration. The biodistribution studies showed that SOD-enzymosomes had a similar organ distribution profile to liposomes with SOD encapsulated in their aqueous interior (SOD-liposomes). SOD-enzymosomes showed earlier therapeutic activity than both SOD-liposomes and free SOD in rat adjuvant arthritis. SOD-enzymosomes, unlike SOD-liposomes, have a therapeutic effect, decreasing liver damage in a rat liver ischemia/reperfusion model. CONCLUSIONS: SOD-enzymosomes were shown to be a new and successful therapeutic approach to oxidative stress-associated inflammatory situations/diseases.

Trace element concentrations in the top predator jumbo squid (Dosidicus gigas) from the Gulf of California.

Jumbo (or Humboldt) squid, Dosidicus gigas, is a large jet-propelled top oceanic predator off the Eastern Pacific. The present study reports, for the first time, concentrations of V, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Cd and Pb in gills, mantle and digestive gland of this powerful pelagic squid in the Gulf of California. Zinc and Cu were the most abundant elements. All elements, with the exception of As, were largely stored in digestive gland; particularly Cd that reached concentrations between 57 and 509 µg g(-1). Significant relationships between tissues were found for Co (digestive gland-gills), As (gills-mantle) and Cd (digestive gland-mantle). Proportionality of Cd concentrations between mantle and digestive gland suggested that detoxification capacity by digestive gland was insufficient to avoid the transfer of this element to mantle and other tissues. Nonetheless, Cd concentrations in the mantle were always below the regulatory limit and, therefore lack of constraints for human consumption. On the basis of the fishery landings, one may estimate that up to 1t of Cd can be annually removed by jumbo squid fisheries.

Partitioning of paralytic shellfish toxins in sub-cellular fractions of the digestive gland of the cockle Cerastoderma edule: Changes under post-bloom natural conditions.

Concentrations of paralytic shellfish toxins (C1+2, B1, dcGTX2+3, dcSTX, GTX2+3 and STX) were determined by LC-FLD in composite samples of digestive glands of the cockle Cerastoderma edule and in each sub-cellular particulate fractions obtained after differential centrifugation (nuclei+debris, mitochondria, lysosomes and microsomes). The specimens were sampled during the exposure to a bloom of Gymnodinium catenatum (day 0) and in the subsequent 8, 12, 14, 19, 21 and 25 days under natural depuration conditions. Toxin profiles of digestive glands were dominated by C1+2 followed by B1 and dcGTX2+3, although the proportion between C1+2 and B1 contents decreased with the time, indicating a slower elimination of B1. All toxins, except GTX2+3 and STX, were quantified in the four sub-cellular fractions. The content of the quantified toxins decreased most markedly in nuclei+debris and microsomal fractions, during the first eight and 12 days, respectively. Conversely, different patterns were observed among toxins in mitochondrial and lysosomal fractions. The less accentuated decreases of dcGTX2+3 and dcSTX contents in the mitochondrial fraction may have resulted from the conversion of other toxins, like C1+2 and B1, associated with enzymatic activities in that fraction. The largest discrepancy was registered in lysosomal fraction for B1, since its content increased after eight days of post-bloom conditions. Input of B1 may come from the conversion of other toxins, like the abundant B2 and C1+2. These transformations are associated to the major role of lysosomes in the intra-cellular digestive process of materials acquired through vesicular transport.

Removal of chromium(III) from contaminated waters using cobalt ferrite: how safe is remediated water to aquatic wildlife?

The release of hazardous elements by industrial effluents to aquatic ecosystems is a potential threat to the environment. Chromium (Cr) is one of the elements whose levels in several freshwater ecosystems should be reduced to promote water reuse. In recent years, magnetic materials have gained increasing interest as sorbents because of their easy removal from treated water through magnetic separation. In this study, colloidal cobalt ferrite (CoFe2O4) particles were investigated as magnetic sorbents for chromium-aqueous chemical species. The oxidative stress responses of Mytilus galloprovincialis mussels exposed to 200 µg/L of Cr, resembling remediated water, were evaluated. More than 95% of Cr was removed from contaminated solutions by CoFe2O4 aqueous suspensions at pH 6 and pH 10. The kinetics of sorption experiments were examined using pseudo-1st order, pseudo-2nd order and Elovich models to evaluate which mathematical model has a better adjustment to the experimental data. The present study revealed that the levels of Cr that remained in remediated water induced limited biochemical changes in mussels, being considered safe for aquatic systems. Overall, the use of cobalt ferrite-based sorbents may constitute a promising approach to remediate contaminated water.

Trace elements in relation to trophic ecology of long-distance migratory shorebirds and seabirds in West Africa.

Shorebirds and seabirds are abundant predators in coastal habitats worldwide, relying upon a high diversity of benthic invertebrates and fish, respectively. While occupying different trophic guilds, they are differently exposed to element contamination entering the coastal food web. Therefore, these birds have been used as bioindicators of environmental contamination in marine ecosystems. We analysed the concentration of trace elements in blood samples of 16 shorebird and seabird migratory species in a major non-breeding site, the Bijagós Archipelago, in regard to their trophic ecology. Overall, our study shows low exposure of this bird community to toxic elements, except for Hg. Most species presented Hg burdens within the moderate toxicity threshold, but one species (Dunlin) presented values at a potential high Hg toxicity risk. We found a positive relationship between Se and Ni concentrations and δ15N values (a proxy for trophic level). In addition, a positive relationship was found between δ13C (a proxy for habitat characteristics) and Hg, Fe, Cu and Mn, while the opposite pattern was recorded for As. Differences were also shown for several trace elements between the two studied trophic guilds: concentrations of As, Pb and Se were higher in pelagic fish consumers (seabirds), whereas Cr, Fe and Sr burdens were higher in benthic invertebrate consumers (shorebirds). Although previous studies in the same site revealed very high concentrations of Cd and Pb in some of the prey species of shorebirds and seabirds (bivalves and fishes), values found in birds for these two elements suggest no toxicological risk. Thus, exposure to toxic elements is not currently a threat to coastal bird populations, namely those facing apparent local declines in Guinea-Bissau, one of the most important non-breeding quarters for of West Africa. Still, Hg burdens were high in some species, deserving further monitoring.

Algal sorbents and prospects for their application in the sustainable recovery of rare earth elements from E-waste.

Efficient and sustainable secondary sourcing of Rare-Earth Elements (REE) is essential to counter supply bottlenecks and the impacts associated with primary mining. Recycled electronic waste (E-waste) is considered a promising REE source and hydrometallurgical methods followed by chemical separation techniques (usually solvent extraction) have been successfully applied to these wastes with high REE yields. However, the generation of acidic and organic waste streams is considered unsustainable and has led to the search for "greener" approaches. Sorption-based technologies using biomass such as bacteria, fungi and algae have been developed to sustainably recover REE from e-waste. Algae sorbents in particular have experienced growing research interest in recent years. Despite its high potential, sorption efficiency is strongly influenced by sorbent-specific parameters such as biomass type and state (fresh/dried, pre-treatment, functionalization) as well as solution parameters such as pH, REE concentration, and matrix complexity (ionic strength and competing ions). This review highlights differences in experimental conditions among published algal-based REE sorption studies and their impact on sorption efficiency. Since research into algal sorbents for REE recovery from real wastes is still in its infancy, aspects such as the economic viability of a realistic application are still unexplored. However, it has been proposed to integrate REE recovery into an algal biorefinery concept to increase the economics of the process (by providing a range of additional products), but also in the prospect of achieving carbon neutrality (as large-scale algae cultivation can act as a CO2 sink).

Microplastics in marine mussels, biological effects and human risk of intake: A case study in a multi-stressor environment.

This study documented seasonal levels of microplastics (MPs) and biomarkers (condition index, neurotoxicity, energy, oxidative stress) in mussels (Mytilus galloprovincialis), and water physico-chemical parameters in the Douro estuary (NE Atlantic coast), and estimated the human risk of MP intake (HRI) through mussels. Mussel stress was determined through the Integrated Biomarker Response (IBR). HRI was estimated from mussel MP concentrations and consumer habits. MPs were mainly micro-fibres (72 %) with varied chemical composition. Seasonal MP means (±SEM) in mussels ranged from 0.111 ± 0.044 (spring) to 0.312 ± 0.092 MPs/g (summer). Seasonal variations of mussel stress (IBR: 1.4 spring to 9.7 summer) and MP concentrations were not related. MeO-BDEs, PBDEs, temperature, salinity and other factors likely contributed to mussel stress variation. HRI ranged from 2438 to 2650 MPs/year. Compared to the literature, MP contamination in mussels is low, as well as the human risk of MP intake through their consumption.

Impact of remobilized contaminants in Mytilus edulis during dredging operations in a harbour area: bioaccumulation and biomarker responses.

Dredging operations in harbours are recurrent to maintain accessibility and navigational depths. One of the main environmental risks of these operations is the remobilization of contaminants trapped in the sediments, rendering them more bioavailable to the biota. However, regulatory policies regarding the contamination risk of dredging chiefly apply to the disposal of dredged materials rather than the direct impact of the procedure itself. In order to assess the ecotoxicological risk of harbour dredging operations in a polluted estuary (the Tagus, W Portugal), the present study compared bioaccumulation and biomarker responses in field-deployed mussels before and after the beginning of operations, complemented by sediment characterization and risk analysis based on standardized sediment quality guidelines. The results revealed a very significant increase in genotoxicity and oxidative stress from the beginning of dredging onwards, which was accompanied by increased bioaccumulation of toxicants, especially polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs). Overall, the results indicate the importance of surveying the direct impacts of these procedures on local contamination, especially considering these sediments had been previously classified as "trace contaminated", according to normative guidelines, and therefore safe for disposal. This study shows the importance of obtaining both chemical and biological data in standard monitoring procedures and that the remobilization of contaminants by dredging operations may be grossly underestimated, which calls for caution when assessing the impact of these activities even in low to moderately polluted areas.

Hepatic proteome changes in Solea senegalensis exposed to contaminated estuarine sediments: a laboratory and in situ survey.

Assessing toxicity of contaminated estuarine sediments poses a challenge to ecotoxicologists due to the complex geochemical nature of sediments and to the combination of multiple classes of toxicants. Juvenile Senegalese soles were exposed for 14 days in the laboratory and in situ (field) to sediments from three sites (a reference plus two contaminated) of a Portuguese estuary. Sediment characterization confirmed the combination of metals, polycyclic aromatic hydrocarbons and organochlorines in the two contaminated sediments. Changes in liver cytosolic protein regulation patterns were determined by a combination of two-dimensional electrophoresis with de novo sequencing by tandem mass spectrometry. From the forty-one cytosolic proteins found to be deregulated, nineteen were able to be identified, taking part in multiple cellular processes such as anti-oxidative defence, energy production, proteolysis and contaminant catabolism (especially oxidoreductase enzymes). Besides a clear distinction between animals exposed to the reference and contaminated sediments, differences were also observed between laboratory- and in situ-tested fish. Soles exposed in the laboratory to the contaminated sediments failed to induce, or even markedly down-regulated, many proteins, with the exception of a peroxiredoxin (an anti-oxidant enzyme) and a few others, when compared to reference fish. In situ exposure to the contaminated sediments revealed significant up-regulation of basal metabolism-related enzymes, comparatively to the reference condition. Down-regulation of basal metabolism enzymes, related to energy production and gene transcription, in fish exposed in the laboratory to the contaminated sediments, may be linked to sediment-bound contaminants and likely compromised the organisms' ability to deploy adequate responses against insult.

Environmental quality assessment combining sediment metal levels, biomarkers and macrobenthic communities: application to the Óbidos coastal lagoon (Portugal).

Macroinvertebrate benthic communities are one of the key biological components considered for the assessment of benthic integrity in the context of the Water Framework Directive (WFD). However, under moderate contamination scenarios, the assessment of macrobenthic alterations at community level alone could be insufficient to discriminate the environmental quality of coastal and transitional waters. Keeping this in view, sediment quality of moderately contaminated sites in a coastal lagoon (Óbidos lagoon, Portugal) was assessed by the combination of sediment metal levels, Carcinus maenas biomarkers (accumulated metals and oxidative stress responses) and macrobenthic communities. Two sites were selected in confined inner branches (BS and BB) and a third one in the middle lagoon (ML). The site BB presented slightly higher levels of metals in sediment but biological variables calculated for macrobenthic data were not significantly different between sites. The biotic index M-AMBI that is being applied to assess environmental quality of transitional waters in the scope of the WFD pointed either to high (site ML) or good quality status (BS and BB) in the selected sites. However, crabs from BB site presented significantly higher levels of Ni in hepatopancreas than those from ML and macrobenthic community structure was significantly different between BB and ML. Additionally, spatial differences were obtained for oxidative stress parameters suggesting that BB site presented stressors for crabs (higher GST and lower GSH(t) at BB site). Factor analysis (PCA) integrating sediment contamination, biomarkers in crabs and macrobenthic data also distinguished BB site as the most environmentally disturbed. On the other hand, at ML site, some macrobenthic variables (equitability and polychaetes' diversity) were found to be enhanced by current environmental conditions, suggesting the existence of a better sediment quality. Current results pointed to the usefulness of integrating macrobenthic community alterations with responses at organism level (bioaccumulation and biochemical endpoints) in order to increase the accuracy of environmental quality assessment in lagoon systems. Moreover, the application of different statistical methods was also found to be recommendable.

Linking trophic ecology with element concentrations in a coastal fish community of the Bijagós Archipelago, West Africa.

We report the concentration of 13 elements in the muscle and liver of 17 coastal fish species of the Bijagós Archipelago, Guinea-Bissau, and link element concentrations to trophic ecology as assessed by carbon (δ13C) and nitrogen (δ15N) stable isotopes. We found higher concentrations of Hg, Fe, Mn, Cu, Zn, Se and As in liver as compared to muscle tissue, and the opposite pattern for Sr and Ca in all fish species. The concentration of Hg and Se in muscle samples increased significantly with δ15N, suggesting a biomagnification of these elements in this food chain. The concentrations of Ca and Sr, Fe and Cr, Ca and Mn, and Fe and Mn were positively correlated to each other in more than 50% of the studied species. Fish constitute the most important animal protein source for people in Guinea-Bissau, and thus assessing the concentrations of potentially toxic elements is relevant for human health.

Assessment of the genotoxic potential of contaminated estuarine sediments in fish peripheral blood: laboratory versus in situ studies.

Juvenile Senegalese soles (Solea senegalensis) were exposed to estuarine sediments through 28-day laboratory and in situ (field) bioassays. The sediments, collected from three distinct sites (a reference plus two contaminated) of the Sado Estuary (W Portugal) were characterized for total organic matter, redox potential, fine fraction and for the levels of metals, polycyclic aromatic hydrocarbons (PAHs) and organochlorines, namely polychlorinated biphenyls (PCBs) and dichloro diphenyl tricholoethane plus its main metabolites (DDTs). Genotoxicity was determined in whole peripheral blood by the single-cell gel electrophoresis (SCGE or "comet") assay and by scoring erythrocytic nuclear abnormalities (ENA). Analysis was complemented with the determination of lipid peroxidation in blood plasma by the thiobarbituric acid reactive substances (TBARS) protocol and cell type sorting. The results showed that exposure to contaminated sediments induced DNA fragmentation and clastogenesis. Still, laboratory exposure to the most contaminated sediment revealed a possible antagonistic effect between metallic and organic contaminants that might have been enhanced by increased bioavailability. The laboratory assay caused a more pronounced increase in ENA whereas a very significant increase in DNA fragmentation was observed in field-tested fish exposed to the reference sediment, which is likely linked to increased lipid peroxidation that probably occurred due to impaired access to food. Influence of natural pathogens was ruled out by unaltered leukocyte counts. The statistical integration of data correlated lipid peroxidation with biological variables such as fish length and weight, whereas the genotoxicity biomarkers were more correlated to sediment contamination. It was demonstrated that laboratory and field bioassays for the risk assessment of sediment contamination may yield different genotoxicity profiles although both provided results that are in overall accordance with sediment contamination levels. While field assays may provide more ecologically relevant data, the multiple environmental variables may produce sufficient background noise to mask the true effects of contamination.

Transcriptomic analyses in a benthic fish exposed to contaminated estuarine sediments through laboratory and in situ bioassays.

The transcription of contaminant response-related genes was investigated in juvenile Senegalese soles exposed to sediments from three distinct sites (a reference plus two contaminated) of a Portuguese estuary (the Sado, W Portugal) through simultaneous 28-day laboratory and in situ bioassays. Transcription of cytochrome P450 1A (CYP1A), metallothionein 1 (MT1), glutathione peroxidase (GPx), catalase (CAT), caspase 3 (CASP3) and 90 kDa heat-shock protein alpha (HSP90AA) was surveyed in the liver by real-time PCR. CASP3 transcription analysis was complemented by surveying apoptosis through the TUNEL reaction. After 14 days of exposure, relative transcription was either reduced or decreased in fish exposed to the contaminated sediments, revealing a disturbance stress phase during which animals failed to respond to insult. After 28 days of exposure all genes' transcription responded to contamination but laboratory and in situ assays depicted distinct patterns of regulation. Although sediments revealed a combination of organic and inorganic toxicants, transcription of the CYP1A gene was consistently correlated to organic contaminants. Metallothionein regulation was found correlated to metallic and organic xenobiotic contamination in the laboratory and in situ, respectively. The transcription of oxidative stress-related genes can be a good indicator of general stress but caution is mandatory when interpreting the results since regulation may be influenced by multiple factors. As for MT1, HSP90 up-regulation has potential to be a good indicator for total contamination, as well as the CASP3 gene, even though hepatocyte apoptosis depicted values inconsistent with sediment contamination, showing that programmed cell death did not directly depend on caspase transcription alone.