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- Minor and trace elements in the shell of Patella aspera (Röding 1798)Publication . Cravo, Alexandra; Foster, P.; Bebianno, Maria JoãoSpecimens of the limpet Patella aspera were collected from a clean, coastal marine site and a contaminated estuary on the south coast of Portugal. The shells were analysed individually for their minor (Mg, Sr) and trace element (Fe, Mn, Zn) content. Mean concentrations of these elements in the shell of P. aspera were 4651, 1318, 35.8, 29.9 and 5.5 μg g−1, respectively. The elemental concentrations exhibited both a marked intra- and inter-population variability. Despite the variability within individual populations, significant differences in the trace element composition were apparent between the shells taken from the two sites. Small shells (<2 g) provided the best resolution between sites for both manganese and iron. Differences in zinc were best resolved for larger shells. The shell of P. aspera has an extraordinarily high magnesium concentration, which is insensitive to gross salinity differences, and a trace metal assemblage that can be interpreted in terms of environmental exposure. On these grounds, it is recommended that the shell of P. aspera is a tissue for potential use in environmental trace metal monitoring.
- A multibiomarker approach in Mytilus galloprovincialis to assess environmental qualityPublication . Cravo, Alexandra; Lopes, Belisandra; Serafim, M.A.; Company, Rui; Barreira, Luísa; Gomes, Tânia; Bebianno, Maria JoãoA multibiomarker approach was carried out for the first time in the South Portuguese Coast using Mytilus galloprovincialis, to assess environmental quality, establish if there are adverse biological responses associated to different sources of anthropogenic contamination and to determine spatial and seasonal trends. For this purpose the battery of biomarkers selected was: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx total and Se dependent), Cytochrome P450 component system, Glutathione-S-transferase (GST) and acetylcholinesterase (AChE), metallothionein (MT) and lead-delta-aminolevulinic acid dehydratase (ALAD), lipid peroxidation (LPO) and Condition Index (CI) along with the determination of PAHs and metals (Cd, Cu, Ni, Pb and Zn). Results show that despite the levels of both organic and metallic contaminants in these eight spots in the South Coast of Portugal not being particularly high compared with other contaminated/polluted sites worldwide, the selected battery of biomarkers responded efficiently to the environmental changes and allowed an environmental assessment between seasons and sites. Different spatial and seasonal responses were evident along the South Coast of Portugal, meaning that the contamination is not homogeneous. This does not only reflect different competition, origin and intensity of contamination, but also different environmental conditions (e.g. temperature, salinity). Along the South Portuguese Coast site 8 was the most contaminated, while site 2 was considered the least contaminated. Despite environmental factors possibly causing difficulties in the general interpretation of biomarker data, those that better responded to environmental contamination were CYP450, SOD-mit and T-GPx for the summation SigmaPAHs, MT (digestive gland) for metals (especially Cu), ALAD for Pb and LPO for both organic and metallic contamination. These biomarkers were also positively correlated with temperature in summer, revealing this as a more stressful/critical season. In future environmental contamination assessments there is no need to analyse the components b5, P418, NADH and NADPH of phase I MFO system, and MT in the gills, since their responses are not evident.
- 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.
- 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.
- Bioaccumulation of metals in the soft tissue of Patella aspera: application of metal/shell weight indicesPublication . Cravo, Alexandra; Bebianno, Maria JoãoPatella aspera limpets were taken from a marine clean site (MCS) and an estuarine contaminated site (ECS). The concentrations of Fe, Mn, Zn, Cu, Cd, Ni and Co were determined individually over the available size range. Since there were significant differences in the soft tissue weight between populations, but shell characteristics were similar (length and weight), metal/shell weight indices (MSI) were calculated in order to compare populations. For both populations iron, zinc and manganese/shell weight indices were consistently higher, in the order Fe[Zn[Mn than those of Cu, Ni, Co and Cd. The results exhibited a marked intra- and interpopulation variability. The highest intra-variability was observed at ECS, particularly where a strong effect of shell weight upon the metal accumulation in the soft tissue was evident. Due to this effect, the comparison of populations was carried out between three selected shell weight ranges representative of light shells (0.45e1.95 g), intermediate shells (2.95e4.45 g) and heavy shells (5.45e 7.33 g). The results indicate that, in general, MSI at ECS (a sewage contaminated site), except for Cd, was higher than at MCS possibly reflecting a higher environmental bioavailability of these metals. The resolution between populations was higher when the smallest range of limpets was considered and it decreased with the increase of shell weight. The highest discrimination amongst populations was found for zinc/shell weight index that showed much higher values than those of Mn, Cu, Cd, Ni and Co/shell weight indices, and spatial differentiation between populations persisted amongst the selected shell weight ranges.
- DNA damage as a biomarker of genotoxic contamination in Mytilus galloprovincialis from the south coast of PortugalPublication . Catarina Almeida, Ana; G. Pereira, Catarina; Gomes, Tânia; Bebianno, Maria João; Cravo, AlexandraDNA damage was evaluated in the haemolymph of Mytilus galloprovincialis from nine sites along the south coast of Portugal using the comet assay. DNA damage was low, in the same range of sites considered to suffer low impact from genotoxic contaminants. Even so, differences between sites, seasons and genders were found. Highest values were in mussels from the main estuaries and the fishery harbour, reflecting higher genotoxin levels, whereas the lowest values can be used as a baseline for future work. Non-contaminant related factors (e.g. temperature and oxygen) were also shown to influence DNA damage. Between seasons, highest values were in summer related not only to the increase of tourism in this region (∼10-fold), but also to temperature. Between genders, males were found to be more sensitive. The condition index was also generally higher in summer. Lipid peroxidation, another damage biomarker, was measured in gills to assess if there is any association between the responses of both biomarkers and if they are similarly affected by the same environmental conditions. LPO like DNA damage was higher in summer. This work confirms that DNA damage is a sensitive biomarker to discriminate genotoxic contamination, even in areas considered to suffer low impact from genotoxins.
- A multi-biomarker approach in cross-transplanted mussels Mytilus galloprovincialisPublication . Serafim, M.A.; Lopes, Belisandra; Company, Rui; Cravo, Alexandra; Gomes, Tânia; Serrão Sousa, Vânia; Bebianno, Maria JoãoThe present work integrates the active biomonitoring (ABM) concept in mussels Mytilus galloprovincialis from the South coast of Portugal transplanted during 28 days between two sites with different sources of contamination, and vice versa, in order to assess biological effects in these mussels. For that purpose a multibiomarker approach was used. The suit of biomarkers indicative of metal contamination were metallothioneins (MT) and the enzyme δ-aminolevulinic acid dehydratase (ALAD), for organic contamination mixed function oxidase system (MFO), glutathione-S-transferase (GST) and acetylcholinesterase (AChE), as oxidative stress biomarkers superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and lipid peroxidation (LPO). These biomarkers were used to determine an index to evaluate the stress levels in these two sites. Site A is strongly influenced by metallic contamination, with higher Cu, Cr and Pb in M. galloprovincialis, as well as higher MT levels, antioxidant enzymes activities and LPO concentrations, and lower ALAD activity. In site B organic compounds (PAHs) are prevalent and native mussels show higher activities of the MFO system components and GST. Transplanted mussels had significant alterations in some biomarkers that reflect the type of contaminants present in each site, which demonstrates the primary role of the environment in determining the physiological characteristics of resident mussels. Therefore the application of ABM using a battery of biomarkers turns out to be a useful approach in sites where usually complex mixtures of contaminants occurs. In this study the biomarkers that better differentiate the impact of different contaminants at each site were MT, CYP450, SOD and CAT.
- Temporal variability of the mass exchanges between the main inlet of Ria Formosa lagoon (southwestern Iberia) and the Atlantic OceanPublication . Rosa, Alexandra; Cardeira, Sara; Pereira, Catarina; Rosa, Monica; Madureira, Pedro Miguel; Rita, Filomena; Jacob, J.; Cravo, AlexandraUnderstanding the mass exchange patterns between coastal lagoons and ocean plays a key role to explain their impact upon the water quality and biological productivity of these systems. Ria Formosa is a temperate lagoon on the southwestern coast of Iberia, one of the most important coastal system in this region in terms of biological productivity, ecological and economic values. Given its relevance and strong interconnectivity with the adjoining ocean, the quantification of these exchanges is a key issue that had not yet been thoroughly addressed. In this context, this study is focused on understanding the role of Ria Formosa's main inlet in terms of mass budgets dynamics of water, nutrients, suspended solids and chlorophyll a with the Atlantic Ocean and to identify its seasonal variability. In order to attain this purpose, the influence of the forcing mechanisms at different time scales, including tides, oceanographic/meteorological synoptic conditions and seasons, was assessed. To accomplish this, six semidiurnal tidal cycles surveys were conducted at Faro-Olhao inlet, comprising hourly water samples collection and in situ measurements at a selected cross-section of the inlet channel. Results revealed that mass exchanges variability through Faro-Olhao inlet was mainly due to oceanographic processes (upwelling and coastal countercurrent events) and, secondly, to phytoplankton activity within the lagoon. Seasonally, regardless the direction of the residual current through the inlet, Ria Formosa acted as a source of material during Spring and Summer seasons, which contributed to increase the biological productivity of the coastal ocean. Upwelling events that occurred more evidently during the Autumn survey drove an import amount of nutrients into the lagoon, enhancing its biological productivity. Furthermore, the rainfall period that prevailed before the Winter survey contributed to export material and nutrients to the adjacent ocean, confirming that Ria Formosa fertilizes the coastal ocean, even during a period of low productivity.
- Variability of nutrients and chlorophyll a in Ancao Inlet during a spring tidal cycle in April 2009Publication . Alcantara, P.; Cravo, A.; Jacob, J.; Cravo, AlexandraSao Luis or Ancao Inlet is the most western of the six inlets of the Ria Formosa lagoon, in southern Portugal. This inlet contributes relatively little to the water volume exchange over a tidal cycle (similar to 10%). Temperature, salinity, pH, dissolved oxygen, and chlorophyll a and nutrient concentrations were measured hourly at the surface, at an intermediate level, and near the bottom over a complete spring tidal cycle, at a sampling point in the middle of the inlet. Current velocity was also measured hourly over the entire cross-section of the inlet using an acoustic Doppler current meter. From these parameters, the flow rate and the transport of nutrients and chlorophyll a were calculated and integrated over the complete tidal cycle providing the net transport. The results show that this is a well-mixed dynamic inlet where, during spring tide, the tidal signal is evident particularly for the nutrient and chlorophyll a concentrations. The nutrient concentrations were low (almost depleted), while the chlorophyll a concentrations were high (4-6 mg m(-3)). The estimated transports through Ancao Inlet revealed that there was a net import of chlorophyll a, nitrate, and phosphate to the Ria Formosa lagoon, while silicate was exported to the adjacent coastal area.
- Using biochemical and isotope geochemistry to understand the environmental and public health implications of lead pollution in the lower Guadiana River, Iberia: A freshwater bivalve studyPublication . Company, Rui; Serafim, Angela; Lopes, Belisandra; Cravo, A.; Shepherd, T. J.; Pearson, G.; Bebianno, Maria João; Cravo, AlexandraLead is a natural component of aquatic ecosystems with no known biological role and is highly toxic. Its toxicity stems from its ability to mimic biologically important metals and to produce membrane damage through lipid peroxidation (LPO). Most lead poisoning symptoms are thought to occur by interfering with an essential enzyme, delta-aminolevulinic acid dehydratase (ALAD), the activity of which is markedly inhibited by lead. The purpose of this work was to study the levels and effects of lead pollution (responses of ALAD and oxidative stress biomarker LPO) in the freshwater bivalve Corbicula fluminea along the lower Guadiana River (Portugal and Spain); a major river system impacted by historic mining pollution and more recent anthropogenic inputs. The results show that the enzymatic activity of ALAD is negatively correlated with the total Pb concentration of the whole tissue suggesting that ALAD has considerable potential as a biomarker of lead exposure in C. fluminea. To identify the sources of lead to which bivalves have been exposed, high precision (206)Pb/(204)Pb, (207)Pb/(204)Pb, (208)Pb/(204)/Pb ratios for C. fluminea confirm that historical mining activities in the Iberian Pyrite Belt are the dominant source of lead pollution in the lower Guadiana River. The isotope patterns however exhibit marked seasonal and geographic variation in response to rainfall and river water management. Locally, other anthropogenic sources of lead have been detected in C. fluminea close to population centres, thus adding to its versatility as a freshwater bio-indicator. Overall, the study highlights the value of natural ecosystems as monitors of water quality and their importance for public health assessment and surveillance.