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- 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.
- 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.
- 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.
- Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis)Publication . Serafim, Angela; Lopes, Belisandra; Company, Rui; Ferreira, A. M.; Bebianno, Maria JoãoAliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed. Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2-3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.
- Induction of cadmium-binding peptides in sorghumPublication . Pinto, Ana Paula Honrado; Serafim, Angela; Bebianno, Maria; Almeida Mota, Ana MariaHydroponic experiments with sorghum were carried out in nutrient solution medium, under controlled environmental conditions for different periods of time. Cadmium (Cd ) was added at a final concentration of 0.1, 1 and 10 mg dm−3. The effect of time and cadmium concentration on the content of thiolic groups (TSH) in the “cell sap” of sorghum tissues are discussed in relation to plant resistance/tolerance to metal stress. The increase in Cd level in the nutrient solution was associated with a higher total Cd accumulation in the plant (root and shoot), which produced a significant enhancement on the thiolic groups content. Furthermore, higher Cd concentrations in plant tissues induced not only an increase in sulphur-rich compounds but also enhanced the binding capacity of these compounds.
- Response of antioxidant systems to copper in the gills of the clam Ruditapes decussatusPublication . Geret, Florence; Serafim, Angela; Barreira, Luísa; Bebianno, Maria JoãoCopper (Cu) is an essential element for biological systems, however, when present in excess, is toxic. Metallothioneins can play an important role in Cu homeostasis and detoxification. Moreover, Cu can catalyse the production of toxic hydroxyl radicals that cause lipid peroxidation but defence systems in the cells can limit the oxidative damage. The present study was performed to investigate the effect of three Cu concentrations (0.5, 2.5 and 25 mug l(-1)) on the response of antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathion peroxidase and total glutathion peroxidase), total proteins, metallothioneins (MT), malondialdehyde (MDA) concentrations in the gills of the clam, Ruditapes decussatus. The activity of antioxidant enzymes and total proteins, MT and MDA concentrations were measured in the gills of the clams after 1, 3, 7, 14, 21 and 28 days of Cu exposure. Results indicate that Cu only induces an imbalance in the oxygen metabolism during the first week of Cu exposure due to a decrease in mitochondrial SOD and CAT, selenium-dependent and total glutathion peroxidase activities. Cu also causes lipid peroxidation, measured by the MDA formation, that was Cu dependent. In the gills of clams exposed to 25 mug Cu/l, the excess of Cu triggers the induction of MT synthesis after 3 days. of exposure.
- Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricusPublication . Company, Rui; Serafim, Angela; Bebianno, Maria João; Cosson, R.; Shillito, B.; Fiala-Médioni, A.Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se-GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 microM), Cu (0.4 microM) and Hg (0.1 microM) with different times of exposure. The experiments were performed in pressurized containers at 9+/-1 degrees C and 85 bars. Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.
- Adaptation of the antioxidant defence system in hydrothermal-vent mussels (Bathymodiolus azoricus) transplanted between two Mid-Atlantic Ridge sitesPublication . Company, Rui; Serafim, Angela; Cosson, Richard; Fiala-Medioni, Aline; Dixon, David R.; Bebianno, Maria JoãoThe vent mussel Bathymodiolus azoricus is the dominant member of the Northern Mid-Atlantic Ridge (MAR) hydrothermal megafauna, and lives in an environment characterized by temporal and spatial variations in the levels of heavy metals, methane and hydrogen sulphide, substances which are known to increase reactive oxygen species levels in the tissues of exposed organisms. To evaluate the effects of two contrasting hydrothermal environments on the antioxidant defence system of this vent mussel species, a 2-week transplant experiment was carried out involving mussels collected from the relatively deep (2300 m), and chemical rich, Rainbow vent field. These were transplanted to the shallower (1700 m), and relatively less toxic, Lucky Strike vent field. To achieve this objective, levels of superoxide dismutase, catalase (CAT), total glutathione peroxidase (GPx), selenium-dependent glutathione peroxidase and lipid peroxidation (LPO) were measured in the gills and mantle tissues of resident and transplant mussels before and after the transplant experiment. With the exception of CAT, the gills of the transplanted mussels had significantly higher antioxidant enzyme activity compared with the basal levels in the donor (Rainbow) and recipient (Lucky Strike) populations; whereas the antioxidant enzyme levels in the mantle tissues of the transplants reflected the baseline levels of activity in the native Lucky Strike mussels after 2 weeks. In contrast, LPO levels were significantly higher in both tissue types in the transplants than in either the source or the recipient populations, which suggested a response to hydrostatic pressure change (note, the transplant animals were brought to the surface for transportation between the two vent fields). The fact that the Rainbow mussels survived the transplant experience indicates that B. azoricus has a very robust constitution, which enables it to cope behaviourally, physiologically and genetically with the extreme conditions found in its naturally contaminated deep-sea environment.
- Assessment of Iodine Deficiency in Sao Tome e Principe ChildrenPublication . Serafim, Angela; Martins-Ferreira, Ana; Serafim, P.; Pedro, Paulo; Rodrigues, Celia; Coutinho, Miguel; Figueiredo, TeresaIntroduction: Iodine deficiency still constitutes one of the major public health problems in the world, having a serious social and economic impact. This global problem involves about 130 countries, including Sao Tome and Principe (STP). The aim of the present study was to determine the prevalence and distribution of iodine deficiency in male children within different host institutions of STP. Material and Methods: A descriptive, cross-sectional observational study was conducted with a non-probabilistic sample (for convenience) of 116 children. Children were aged from 6 to 17 years, with an overall mean age of 11.1 +/- 3.2 years. Iodine intake was evaluated in 6 districts of STP, through two indirect indices: (i) urine iodine (UI) determinations (ii) prevalence of goiter evaluated by cervical ultrasound. Results: The global median UI value was 109 mu g/L (P20: 78.2 mu g/Land P80: 140.3 mu g/L), with 16.4% of children <50 mu g/L and 40.5% <100 mu g/L. Considering children aged 6 - 12 years, 15.6% showed U1 <50 mu g / L, while in group aged 13-17 years this percentage increased to 17.9%. These values fulfill WHO criteria of sufficient region in iodine. On the other hand, the goiter prevalence against age was 29.7% and against Body Surface Area (BSA) was 44.6%, which is much higher than the cut-off established by the WHO (5%). Conclusion: The iodine deficiency and endemic goiter in STP are currently a public health problem. Therefore, it will be necessary to ensure that the interventions carried out are efficiently monitored, in order to eradicate endemic goiter, without risks of hyperthyroidism.
- Effect of temperature and size on metallothionein synthesis in the gill of Mytilus galloprovincialis exposed to cadmiumPublication . Serafim, Angela; Company, Rui; Bebianno, Maria João; Langston, W.JAlthough the degree of metal contamination is expected to be a primary determinant of metallothionein (MT) induction in marine mussels, at least at polluted sites, variability caused by temperature, and biotic factors such as size, may need to be considered when interpreting field data. To test the effects of these variables, mussels, Mytilus galloprovincialis, of different sizes (small: 3.5+/-0.5 cm and large: 5.2+/-0.7 cm) were exposed to Cd (100 mug 1(-1)) at different water temperatures (5, 18 and 25 degreesC) for 34 days. Resultant Cd and MT concentrations in gills were shown to be size dependent and increased significantly with temperature. At the highest temperature tested (25 degreesC) there appears to be a disproportionate effect on Cd accumulation, which raises MT synthesis to exceptionally high levels. The effect of size on MT concentrations was also temperature-dependent: at 18 and 25 degreesC, large mussels exhibited higher MT levels than smaller individuals, whilst at 5 degreesC there were no significant differences between contaminated and control mussels, in either size-group, as a result of the reduced level of MT production at this temperature. When considering the use of MT levels in mussels as a biochemical indicator of metal contamination, the potential influence of factors such as size and temperature on MT-metal relationships needs to be considered. Samples should be of uniform size as far as possible, and collection should be limited to a fixed season (avoiding climatic extremes) to ensure that the effects of these factors on baseline levels of MT is minimised.
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