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- Source and impact of lead contamination on δ-aminolevulinic acid dehydratase activity in several marine bivalve species along the Gulf of CadizPublication . Company, Rui; Serafim, M.A.; Lopes, Belisandra; Cravo, Alexandra; Kalman, J.; Riba, I.; DelValls, T. A.; Blasco, J.; Delgado, J.; Sarmiento, A . M.; Nieto, J. M.; Shepherd, T. J.; Nowell, G.; Bebianno, Maria JoãoCoastal areas and estuaries are particularly sensitive to metal contamination from anthropogenic sources and in the last few decades the study of space-time distribution and variation of metals has been extensively researched. The Gulf of Cadiz is no exception, with several rivers draining one of the largest concentrations of sulphide deposits in the world, the Iberian Pyrite Belt (IPB). Of these rivers, the Guadiana, one of the most important in the Iberian Peninsula, together with smaller rivers like the Tinto and Odiel, delivers a very high metal load to the adjacent coastal areas. The purpose of this work was to study the source and impact of lead (Pb) drained from historical or active mining areas in the IPB on the activity of a Pb inhibited enzyme (δ-aminolevulinic acid dehydratase, ALAD) in several bivalve species along the Gulf of Cadiz. Seven marine species (Chamelea gallina, Mactra corallina, Donax trunculus, Cerastoderma edule, Mytilus galloprovincialis, Scrobicularia plana and Crassostrea angulata) were collected at 12 sites from Mazagón, near the mouth of the rivers Tinto and Odiel (Spain), to Cacela Velha (Ria Formosa lagoon system, Portugal). Lead concentrations, ALAD activity and lead isotope ratios ((206)Pb/(204)Pb, (207)Pb/(204)Pb and (208)Pb/(204)Pb) were determined in the whole soft tissues. The highest Pb concentrations were determined in S. plana (3.50±1.09 μg g(-1) Pb d.w.) and D. trunculus (1.95±0.10 μg g(-1) Pb d.w.), while M. galloprovincialis and C. angulata showed the lowest Pb levels (<0.38 μg g(-1) Pb d.w.). In general, ALAD activity is negatively correlated with total Pb concentration. However this relationship is species dependent (e.g. linear for C. gallina ALAD=-0.36[Pb]+0.79; r=0.837; or exponential for M. galloprovincialis ALAD=2.48e(-8.3[Pb]); r=0.911). This indicates that ALAD activity has considerable potential as a biomarker of Pb and moreover, in marine bivalve species with different feeding habits. Lead isotope data showed significant seasonal and spatial changes in bivalve isotopic composition reflecting seasonal and geographic differences in bioaccumulation. Within the study area, Pb can be modelled as a mixing between geogenic Pb and mine-related, discharges of Pb from the IPB. For some sites at the mouth of the Guadiana River, the bivalves show contamination from other anthropogenic sources, such as leaded boat/aviation fuel and/or leaded paint. Finally, the study demonstrates convincingly the need to consider species-specific variation when using bivalve ALAD activity as a biomarker for Pb.
- Recent evolution of the tidal prisms at the inlets of the western sector of the Ria Formosa, south coast of PortugalPublication . Jacob, J.; Cravo, AlexandraThe Ria Formosa is a coastal lagoon comprising six tidal inlets. The western sector encompassing three inlets, Ancao, Faro-Olhao and Armona is the most important one since it represents approximately 90% of the total tidal prism. The Ancao inlet is characterized by a cyclic eastward migration pattern, but until now the knowledge of the real impact of its dynamics in the last 20 years in terms of tidal prisms and interconnectivity between these three inlets is scarce. This work presents the most exhaustive data set acquired so far, comprising 25 field surveys led in the period 2004-2013 on the western sector of Ria Formosa. This provides a long-term analysis of the recent temporal evolution of the tidal prisms from the three inlets during a single migration cycle (18 years) of the Ancao inlet. It determines the relative contribution of the three inlets to the total prism of the western sector of Ria Formosa before the last artificial opening of the Ancao inlet. Results show that at Ancao inlet the tidal prism decreased in spring tide mainly due to its eastward migration. The other natural inlet, Armona inlet, depict a progressive narrowing responsible for the decrease of the tidal prism in neap tide while during the same tidal phase, at the Faro-Olhao inlet, the most important one in terms of exchanges, the tidal prism increased. Those alterations at the three inlets could be related to morphodynamic changes and morphological evolution and confirmed the strong interconnectivity between them to compensate the variations of their relative contributions to the total prism. The tidal prisms errors were low for all cases (< 10%) and confirm that, globally, there was an increase of the prisms with time.
- Role of the Ria Formosa inlets on the physical, chemical and biological exchanges with the adjoining oceanPublication . Cravo, Alexandra; Jacob, J.A coastal lagoon is a “shallow coastal water body separated from the ocean by a barrier, connected at least intermittently to the ocean by one or more restricted inlets” (Kjerfve, 1994). Coastal lagoons are typically found along low-lying coastlines, affected by a tidal range < 4 m and generally < 5 m deep (Bird, 1994; Kjerfve,1994). Coastal lagoons are important ecosystems because these support a wide range of natural services, highly valuable for society. These complex systems provide food, storm protection, tourism, among others. So, they contribute to the overall productivity of coastal waters by sustaining a variety of habitats, including salt marshes, seagrasses, and/or mangroves particularly important for many fish and shellfish species. Water quantity and quality in a lagoon is influenced by the rate at which the lagoon loses or gains water from exchange with the ocean, surface runoff, evaporation, precipitation and groundwater (Allen et al., 1981).
- 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.
- Integrated approach to assess ecosystem health in harbor areasPublication . Bebianno, Maria João; Pereira, C.; Rey, F.; Cravo, Alexandra; Duarte, D. N.; D'Errico, G.; Regoli, F.Harbors are critical environments with strategic economic importance but with potential environmental impact: health assessment criteria are a key issue. An ecosystem health status approach was carried out in Portimão harbor as a case-study. Priority and specific chemical levels in sediments along with their bioavailability in mussels, bioassays and a wide array of biomarkers were integrated in a biomarker index (IBR index) and the overall data in a weight of evidence (WOE) model. Metals, PAHs, PCBs and HCB were not particularly high compared with sediment guidelines and standards for dredging. Bioavailability was evident for Cd, Cu and Zn. Biomarkers proved more sensitive namely changes of antioxidant responses, metallothioneins and vittellogenin-like proteins. IBR index indicated that site 4 was the most impacted area. Assessment of the health status by WOE approach highlighted the importance of integrating sediment chemistry, bioaccumulation, biomarkers and bioassays and revealed that despite some disturbance in the harbor area, there was also an impact of urban effluents from upstream.
- Genotoxicity of copper oxide and silver nanoparticles in the mussel Mytilus galloprovincialisPublication . Gomes, Tânia; Araújo, Olinda; Pereira, Rita; Catarina Almeida, Ana; Cravo, Alexandra; Bebianno, Maria JoãoThough there is some information on cytotoxicity of copper nanoparticles and silver nanoparticles on human cell lines, there is no information on their genotoxic and cytotoxic behaviour in bivalve molluscs. The aim of this study was to investigate the genotoxic impact of copper oxide and silver nanoparticles using mussels Mytilus galloprovincialis. Mussels were exposed to 10 μg L⁻¹ of CuO nanoparticles and Cu²⁺ and Ag nanoparticles and Ag⁺ for 15 days to assess genotoxic effects in hemocytes using the comet assay. The results obtained indicated that copper and silver forms (nanoparticles and ionic) induced DNA damage in hemolymph cells and a time-response effect was evident when compared to unexposed mussels. Ionic forms presented higher genotoxicity than nanoparticles, suggesting different mechanisms of action that may be mediated through oxidative stress. DNA strand breaks proved to be a useful biomarker of exposure to genotoxic effects of CuO and Ag nanoparticles in marine molluscs.
- Development of a multiresidue method for the determination of 24 pharmaceuticals in clams by QuEChERS and liquid chromatography-triple quadrupole tandem mass spectrometryPublication . Rodrigues, Joao; Albino, Stephanie; Silva, Sofia; Cravo, Alexandra; Cardoso, Vitor Vale; Benoliel, Maria Joao; Almeida, Cristina M. M.Data on different therapeutic classes of pharmaceutical compounds (PhCs) in clams or other bivalves living in natural conditions are scarce. The aim of this work was the optimization and validation of a method for the determination of PhCs in clams for further evaluation of any potential human exposure risk due to their consumption. A quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach is proposed for sample clean-up and concentration of 24 PhCs in clams, with subsequent analysis by liquid chromatography-tandem mass spectrometry. This method showed a good linear range for all PhCs with determination coefficients (r(2)) between 0.9949 and 0.9993 and coefficients of variation (CVm) lower than 5.5%. This method allowed the quantification of target compounds at trace concentration levels (ngg(-1)), being the most abundant PhC in clam caffeine. This PhC was detected in more than 70% of samples with concentrations ranging from 0.10 to 12ngg(-1) wet weight.
- Genotoxicity in two bivalve species from a coastal lagoon in the south of PortugalPublication . Catarina Almeida, Ana; G. Pereira, Catarina; Gomes, Tânia; Cardoso, Cátia; Bebianno, Maria João; Cravo, AlexandraDNA damage was evaluated by comet assay in the haemolymph of two bivalve species Ruditapes decussatus and Mytilus galloprovincialis from the Ria Formosa lagoon (south Coast of Portugal). Clams and mussels were sampled from sites close to each other to determine interspecific responses to similar environmental conditions, considering also seasonal and gender differences. Coupled with genotoxic effect, another damage biomarker (lipid peroxidation) was analysed to verify if the conditions that instigate DNA damage can be related with injury to cell membranes. For both species, DNA damage was low, reflecting the low levels of genotoxic contaminants in the lagoon, and no interspecific differences were found. However, seasonal differences were established for both bivalve species, reflecting higher environmental stress in summer. Regarding gender susceptibility, only clams showed differences in percentage of Tail DNA, with females more sensitive to DNA damage than males. Additionally, results for clams point out that factors responsible for LPO may not be the same as those causing genotoxicity. This study demonstrated that DNA damage is a sensitive biomarker to discriminate spatial, temporal and gender differences, being an appropriate biomarker for genotoxicity evaluation even in places of low contamination, such as the Ria Formosa lagoon.
- Interspecific variability of endocrine disruption and oxidative stress in two bivalve species from the Ria Formosa Lagoon (south coast of Portugal)Publication . Pereira, Catarina; Gomes, Tânia; Cardoso, Catia; Almeida, Ana Catarina; Araújo, Olinda; Bebianno, Maria João; Cravo, AlexandraEstuaries are highly productive ecosystems that have been increasingly affected by anthropogenic activities. In this study, the presence of endocrine disrupting compounds (EDCs) and their relation to oxidative stress were assessed in two bivalve species (mussels Mytilus galloprovincialis and clams Ruditapes decussatus) from the Ria Formosa Lagoon (south Portugal). We applied the alkali-labile phosphate (ALP) assay to determine vitellogenin-like protein levels, and the lipid peroxidation (LPO) method to assess oxidative damage. Mussels and clams exhibited the same pattern of ALP concentrations, with no differences between genders and higher levels in winter. R. decussatus showed higher ALP values than M. galloprovincialis, possibly associated with a stronger responsiveness of clams to EDCs or due to the influence of the reproductive cycle. Mussels exhibited a more evident seasonal effect for LPO, indicating that these organisms are more influenced by seasonal fluctuations in the water column compared to the sediment-associated clams. Both species appear to be under the influence of EDC exposure (which could induce Vg-like protein synthesis) and oxidative stressors (leading to lipid peroxidation) in the Ria Formosa Lagoon, particularly in winter, reflecting the presence of different contaminants in Ria Formosa and environmental changes, as well as complex interactions between them.
- Nutrients and chlorophyll-a exchanges through an inlet of the Ria Formosa Lagoon, SW Iberia during the productive season - Unravelling the role of the driving forcesPublication . Cravo, Alexandra; Cardeira, Sara; Pereira, Catarina; Rosa, Monica; Alcantara, Pedro; Madureira, Miguel; Rita, Filomena; Correia, Cátia; Rosa, Alexandra; Jacob, J.Despite the advances and increase of observations of physical and chemical processes to better understand thefunctioning of coastal environments, a large gap still exists in quantifying exchanges and interactions betweenadjacent coastal systems. It is important to bear in mind that the dynamics and mass exchanges depend on thevariability of the driving mechanisms and respond to several time scales (tidal, seasonal and inter-annual). Thegeneral objective of this work was to unravel the role of the driving forces on the nutrients and chlorophyll-aexchanges at the Ancão inlet of Ria Formosa lagoon during the most productive season - spring. This is thesmallest inlet of the western system, the closest to continental interface and the one where migratory patterns areevident. So, the changes along time could be more evident there than at the other two inlets of the westernsector. Ancão inlet will serve as a reference to compare the mass budgets exchanged with the ocean with theother two inlets. Specifically, this work intends to: i) better understand the behaviour of this inlet; ii) its role inthe mass exchanges; and iii) its interplay with phytoplankton productivity during the peak season. In thiscontext, the transport of water, nutrients and chlorophyll-a(proxy of phytoplankton density) were estimatedacross the Ancão inlet section during the spring season. Surveys were conducted under extreme fortnightly tidalconditions during 2007, 2009 and 2012, covering different hydrodynamic, meteorological and environmentalconditions. To estimate the mass transport,field velocities, nutrients and chlorophyll-aconcentrations weremeasured concurrently along its cross section. Results show that although the hydrodynamic and morphologicchanges recorded at this inlet provoked a loss of hydraulic efficiency apparently these did not markedly affectthe mass exchanges during the spring productive season. Globally, the contribution and intensity of the me-teorological/physical, chemical and biological drivers superimposed those affected by tides, reflected in thedifferences between thefive surveys conducted. Upwelling was frequent during the spring season and played akey role on the exchanges and mass transport through the Ancão inlet. Data also show that nutrients behaveddissimilarly; silicate was consistently exported, nitrate mostly imported, dependent on the intensity of upwellingand biological consumption, while phosphate generally followed the direction of residual circulation.Chlorophyll-ashowed a general import except in both 2012 campaigns, when it was exported in small amounts(< 1 kg). Data support that during spring season, particularly in periods when upwelling is relevant, theavailability of nutrients on the coast will promote afterwards the growth of phytoplankton that enters andfertilizes the Ria Formosa, increasing its productivity. The estimated amounts are relative to the smallest inlet ofthe western sector of Ria Formosa. In consequence, it may be anticipated that through the other main inlets fromthis sector the exchanges are intensified and may enhance their contribution to the Ria Formosa fertilization.