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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.
- Effect of different hydrothermal vent conditions in the proteome of vent mussel Bathymodiolus azoricusPublication . Company, Rui; Serafim, A.; Bebianno, Maria João; Cajaraville, M.; Antúnez, O.; Torreblanca, A.The mytilid mussel Bathymodiolus azoricus is the most representative species of north Mid Atlantic Ridge (MAR) hydrothermal vents and is abundantly found at Menez-Gwen, Lucky Strike and Rainbow vent sites.
- Metallothioneins in the clam Ruditapes decussatus: an overviewPublication . Bebianno, Maria João; Serafim, MA; Simes, DinaThe 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.
- DNA damage and lipid peroxidation vs. protection responses in the gill of Dicentrarchus labrax L. from a contaminated coastal lagoon (Ria de Aveiro, Portugal)Publication . Ahmad, I.; Maria, Vera L.; Oliveira, M.; Serafim, Angela; Bebianno, Maria João; Pacheco, M.; Santos, M.A.The present research work aimed to investigate the damage vs. protection responses in gill of European sea bass (Dicentrarchus labrax) captured at a polluted coastal lagoon, Ria de Aveiro (Portugal), as a tool to evaluate the human impacts on environmental health. Damage was assessed as DNA strand breakage and lipid peroxidation (LPO) whereas protection was evaluated by measuring catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), glutathione S-transferase (GST), total glutathione (GSHt), thiols and metallothioneins (MT). Fish were caught at five locations: Gafanha (GAF), Rio Novo do Príncipe (RIO), Laranjo (LAR) and Vagos (VAG) presenting each a different recognized source of contamination, and Torreira (TOR), assumed as reference site. Among the surveyed sites, gill damage was observed only at GAF, as measured by DNA integrity loss and LPO increase. An overall induction in enzymatic antioxidant protection was perceptible in fish from GAF and VAG, expressed as higher GPX, GR and GST activities. In addition, LAR fish showed elevated GST and CAT activities. Thiols content was higher in all study sites irrespective to the pollution spectrum, whereas GSHt increase was only observed at GAF and VAG. The highest MT level was detected in fish from VAG and the lowest level from RIO suggesting a low contamination degree on this particular site. Globally, the results expressed site-specific response patterns, signalling two critical areas - GAF and VAG. Additionally, a joint analysis (damage vs. protection) allowed the following ordering of surveyed sites according to the contamination degree and risk to fish health; GAF>VAG>LAR>RIO>TOR. D. labrax gill responses demonstrated their efficacy as early warning signals of the contaminants presence. Moreover, the adopted approach, considering simultaneously protection responses and damaging effects, also revealed its usefulness on the pollution extent assessment.
- Habitat quality of estuarine nursery grounds: Integrating non-biological indicators and multilevel biological responses in Solea senegalensisPublication . Fonseca, V. F.; Vasconcelos, R. P.; Tanner, S. E.; Franca, S.; Serafim, M.A.; Lopes, Belisandra; Company, Rui; Bebianno, Maria João; Costa, M. J.; Cabral, H. N.Estuaries are highly valuable ecosystems that provide various goods and services to society, such as food provision and supporting nursery habitats for various aquatic species. Estuarine habitat quality assessment is thus critical in managing both ecological and economic value. In this work, various biological and non-biological indicators of habitat quality in estuarine nursery areas were determined, encompassing local environmental conditions, chemical contamination, anthropogenic pressures, juvenile Solea senegalensis condition, biomarkers response to contamination and juvenile density. The various indicators provided an integrated view on habitat quality and their responses were broadly concordant. Nursery quality assessment based on anthropogenic pressure indicators and fish biomarker responses were very similar, signaling nursery areas with higher anthropogenic pressure in Tejo and Ria de Aveiro estuaries. Yet, favorable environmental conditions across all sites could have contributed to lessen the potential hazardous biological effects of exposure to anthropogenic stressors, resulting in soles' fairly good condition and generally high juvenile density. Nevertheless, a mismatch between high juvenile density and high estuarine contribution to adult coastal populations was observed in areas with higher anthropogenic pressures. Although a causal relationship cannot be established, the results emphasize the need to fully understand how the estuarine period spent in estuaries and local processes determine the quantity and quality of juveniles exported to marine adult populations, which is critical to achieve the full potential of the fish production service of estuaries and coastal stock replenishment. (C) 2015 Elsevier Ltd. All rights reserved.
- 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.
- Wild juvenile Dicentrarchus labrax L. liver antioxidant and damage responses at Aveiro Lagoon, PortugalPublication . Maria, Vera L.; Ahmad, I.; Oliveira, M.; Serafim, Angela; Bebianno, Maria João; Pacheco, M.; Santos, M. A.The Aveiro Lagoon, at the north-western coast of Portugal, has been under considerable anthropogenic pressure for the last 5 decades. In order to perform an adequate survey of the effects induced by the contaminants in presence, wild juveniles Dicentrarchus labrax (sea bass) were selected. Thus, sea bass was captured at five sites: Torreira (TOR, as reference site), Gafanha (GAF), Rio Novo Príncipe (RIO), Laranjo (LAR) and Vagos (VAG) in autumn 2005. Liver defence responses such as catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), total glutathione (GSHt), total non-protein thiols (NP-SH) and metallothioneins (MT) were measured. Liver damage was determined as lipid peroxidation (LPO) and DNA integrity loss. RIO, LAR and VAG presented lower CAT, GR, GST activities and NP-SH and GSHt depletion in comparison to TOR. VAG and LAR showed higher GPx activity when compared to TOR. The highest MT level was found at GAF and VAG. The NP-SH and DNA integrity decreased at GAF compared to TOR. This field study demonstrated that not only antioxidant induction but also inhibitory responses must be considered as a signal of contamination.
- Short-term variability of multiple biomarker response in fish from estuaries: Influence of environmental dynamicsPublication . Fonseca, V.F.; França, S.; Vasconcelos, R.P.; Serafim, M.A.; Company, Rui; Lopes, Belisandra; Bebianno, Maria João; Cabral, H.N.Short-term variability in biomarker responses and the effects of temperature and salinity variation were explored in three fish species (Dicentrarchus labrax, Solea senegalensis and Pomatoschistus microps) occurring in the Tejo estuary. Short-term variability in biomarkers was observed in all species although no pattern was discerned over time (days to weeks). Antioxidant enzymes activity (superoxide dismutase (SOD) and catalase (CAT)) showed low temporal variability, indicating some constancy or baseline level in antioxidant responses. Only CAT activity in S. senegalensis was correlated with temperature, suggesting that exposure to contaminants triggered antioxidant acclimation. Higher short-term variability was observed in xenobiotic biotransformation enzymes activity (phase I ethoxyresorufin O-deethylase (EROD) and phase II glutathione S-transferase (GST)). Yet a significant correlation between EROD and GST in D. labrax and S. senegalensis suggests a concomitant response to contaminants. Moreover the lack of correlation between xenobiotic biotransformation enzymes and abiotic variables on concordant time scales, suggest a high specificity of these biomarkers to chemical exposure, rather than high variability due to environmental dynamics.
- Temporal variation in the antioxidant defence system and lipid peroxidation in the gills and mantle of hydrothermal vent mussel Bathymodiolus azoricusPublication . Company, Rui; Serafim, Angela; Cosson, Richard; Fiala-Médioni, Aline; Dixon, David; Bebianno, Maria JoãoHydrothermal vent mussels are exposed continually to toxic compounds, including high metal concentrations and other substances like dissolved sulphide, methane and natural radioactivity. Fluctuations in these parameters appear to be common because of the characteristic instability of the hydrothermal environment. Temporal variation in the antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidases (Total GPx), selenium dependent glutathione peroxidases (Se-GPx)), metallothioneins and lipid peroxidation (LPO) in the gills and mantle of the mussel Bathymodiolus azoricus from Menez-Gwen hydrothermal vent site was evaluated and related to the accumulated metal concentrations (Ag, Cu, Cd, Fe, Mn and Zn) in the tissues. Maximum antioxidant enzyme activities in the gills were detected in the beginning of summer, followed by a gradual decrease throughout the following months. One year after, the levels of antioxidant enzyme activities were similar to those reported one year before. LPO in this tissue exhibited a similar temporal variation trend. A different pattern of temporal variation in antioxidant enzyme activities was observed in the mantle, with a gradual increase from summer to the end of autumn (November). LPO in the mantle exhibited an almost reverse trend of temporal variation to that of antioxidant enzyme activities in this tissue. Antioxidant defences in the gills of B. azoricus were significantly enhanced with increasing concentrations of Ag, Cu and Mn, while negative relationships between antioxidant enzymes and Cd, Cu, Mn and Zn concentrations in the mantle were observed, suggesting different pathways of reactive oxygen species (ROS) production and that these tissues responded differently to the metal accumulation. However, temporal variation in biomarkers of defence and damage were in general similar to coastal bivalve species and can be associated with temporal variations of the physiological status due to reproduction. These variations might also be linked to the highly unstable nature of the hydrothermal environment. (c) 2006 Elsevier Ltd. All rights reserved.
- Sub-lethal effects of cadmium on the antioxidant defence system of the hydrothermal vent mussel Bathymodiolus azoricusPublication . Company, Rui; Serafim, M.A.; Cosson, Richard P.; Fiala-Médioni, Aline; Camus, Lionel; Serrão-Santos, Ricardo; Bebianno, Maria JoãoThe mussel Bathymodiolus azoricus is one of the most abundant species in the Mid-Atlantic Ridge hydrothermal vents and is continually exposed to the high-temperature venting fluids containing high metal concentrations and enriched in sulphides and methane, which constitute a potential toxic environment for marine species. The aim of this study was to assess the effects of a sub-lethal Cd concentration on the antioxidant defence system of this mussel. B. azoricus were collected at Menez Gwen vent site (37 degrees 51'N, 32 degrees 31'W) and exposed to Cd (50 microg l(-1)) during 24 days, followed by a depuration period of six days. A battery of stress related biomarkers including antioxidant enzymes (superoxide dismutase-SOD, catalase-CAT; glutathione peroxidases-GPx), metallothioneins (MT), lipid peroxidation (LPO) and total oxyradical scavenging capacity (TOSC) were measured in the gills and mantle of B. azoricus. Cd was accumulated linearly during the exposure period in both tissues and no significant elimination occurred after the 6 days of depuration. Antioxidant enzymes activities were significantly higher in the gills. Cyt-SOD, T-GPx and Se-GPx were induced during the experiment but this was also observed in control organisms. Mit-SOD and CAT activities remained relatively unchanged. MT levels increased linearly in the gills of exposed mussels in the first 18 days of exposure. No significant differences were observed between LPO levels of control and exposed mussels. TOSC levels remained unchanged in control and exposed mussels. This suggests that although Cd is being accumulated in the tissues of exposed mussels, MT defence system is enough to detoxify the effect of Cd accumulated in the tissues. Furthermore, other factors besides the presence of Cd are influencing the antioxidant defence system in B. azoricus.