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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 JoĂŁo; 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 PrĂncipe (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 JoĂŁo; 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 PrĂncipe (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.
- Biomarkers of damage and protection in Mytilus galloprovincialis cross transplanted in Ria Formosa Lagoon (Portugal)Publication . Maria, Vera L.; Santos, M. A.; Bebianno, Maria JoĂŁoThe study was carried out to check the water contamination in two sites of Ria Formosa Lagoon by assessing the damage responses versus protection in the gills and digestive gland (Dg) of mussels--Mytilus galloprovincialis Lmk. Damage was assessed as DNA integrity and lipid peroxidation (LPO) while protection was evaluated by measuring catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), total glutathione (GSHt) and metallothioneins (MT). Mussels were collected in March of 2007 at Ramalhete (R) as a reference site and Faro port (F) as harbour area. Cross transplantation was from R to F and vice versa performed during 0, 3 and 6 days. Gills DNA integrity decrease was observed in mussels transplanted to F (3 days) and R (6 days). Highest Dg LPO was found in mussels sampled at R. Elevated gills CAT, GPx, GR and GST was observed in mussels transplanted to F. Dg GSHt decreased in mussels transplanted to F. Gills and Dg mussels MT increased in mussels transplanted to R. M. galloprovincialis responses show their capability as early warning signals of the contaminants presence. The adopted approach, considering simultaneously protection responses and damaging effects, also revealed its usefulness on the pollution assessment.
- Antioxidant and lipid peroxidation responses in Mytilus galloprovincialis exposed to mixtures of benzo(a)pyrene and copperPublication . Maria, Vera L.; Bebianno, Maria JoĂŁoThis study aimed to assess the antioxidant system potential and lipid peroxidative effects, in the gill and digestive gland of Mytilus galloprovincialis exposed to individual and binary mixtures of benzo(a)pyrene (BaP) and Cu for 7 days. Data demonstrated that in mussels exposed to BaP antioxidant enzymes (catalase--CAT, total glutathione peroxidase--tGPx, glutathione S-transferase--GST and glutathione reductase--GR) and lipid peroxidation (LPO) increased in the gill. On the contrary, in the digestive gland inhibitory antioxidant effects (superoxide dismutase-SOD, GR, metallothioneins-MT) and no changes in LPO levels were detected. Cu was also a potent oxidant agent since MT and LPO levels increased in mussel gill, despite no LPO effect in the digestive gland. For both single contaminants the organ specificity and distinct physiologic/metabolism roles were evident in terms of antioxidant capacity. Gill SOD inhibition, MT and GST unchanged was a result of "simple independent action" of exposure to BaP and Cu. "Interactions" in the binary mixtures, led to absence of changes in LPO effects. In the digestive gland, BaP and Cu interactions were also responsible for the GST and LPO enhancement (antagonistic effects). The current findings demonstrate the differences in antioxidant responses where the organ dependency highlights each contaminant particular mode of action. Generally, in the gill "non-interactive" effects occurred with the lowest Cu concentration while "interactions" exist for the mixture with the highest Cu concentrations. In the digestive gland, "interactions" and "no interaction" effects occurred in all the binary mixtures. Complex contaminant mixtures interact differently based on target tissue which may lead to an imbalance in the mussels health status.
- Hepatic metallothionein concentrations in the golden grey mullet (Liza aurata): relationship with environmental metal concentrations in a metal-contaminated coastal system in PortugalPublication . Oliveira, M.; Ahmad, I.; Maria, Vera L.; Serafim, A.; Bebianno, Maria JoĂŁo; Pacheco, M.; Santos, M.A.This field survey was designed to assess the environmental metal contamination status of Ria de Aveiro (Portugal). To achieve that goal, the concentrations of Cd, Hg, Cu and Zn in the sediments and water were assessed and Liza aurata hepatic metallothionein (MT) determined. The relationships between MT and environmental metal concentrations and hydrological factors were examined. Results revealed a wide distribution of metals both in water and sediments throughout the lagoon, mainly at Rio Novo do Principe (RIO) and Laranjo (LAR), at concentrations that may affect biota. MT concentrations were higher at the sites with high metal content (RIO and LAR). A significant positive correlation was found between MT and Cd in the sediments as well as with MT and Hg and Cu in the water. Moreover, a negative correlation between MT and salinity was found. Thus, the current data support MT use as a biomarker of metal exposure emphasizing the importance of hydrological parameters in its concentrations. Results suggest the continued monitoring of this lagoon system.
- Comparison of metal accumulation between âArtificial Musselâ and natural mussels (Mytilus galloprovincialis) in marine environmentsPublication . Gonzalez-Rey, Maria; Lau, T. C.; Gomes, TĂąnia; Maria, Vera L.; Bebianno, Maria JoĂŁo; Wu, R.The passive sampler called 'Artificial Mussel' (AM) developed by Wu et al. (2007) can provide a time-integrated estimate of metal concentrations in the marine environment, and offers a potential device to assess and compare metal concentration in different marine environments worldwide. The aim of this study was to compare metal accumulation on AM and natural mussel Mytilus galloprovincialis at three sites with different metal loads along the Portuguese coast for four months. M. galloprovincialis were placed in cages alongside AMs at each site. Samples were collected monthly and Cd, Cr, Cu, Pb, and Zn concentrations in whole soft tissues and AMs compared. For both Cu and Cd, the results were similar between AMs and natural mussels. Higher concentrations of Zn were observed in natural mussels, whereas the inverse was shown for Pb (about 10-fold higher). Our results showed that AMs are promising tools for assessing metal concentrations in marine environments.
- Transcriptomic effects of the non-steroidal anti-inflammatory drug Ibuprofen in the marine bivalve Mytilus galloprovincialis LamPublication . Maria, Vera L.; Amorim, Monica J. B.; Bebianno, Maria JoĂŁo; Dondero, FrancescoThe transcriptomic effects of Ibuprofen (IBU) in the digestive gland tissue of Mytilus galloprovincialis Lam. specimens exposed at low environmental concentrations (250 ng L-1) are presented. Using a 1.7 K feature cDNA microarray along with linear models and empirical Bayes statistical methods 225 differentially expressed genes were identified in mussels treated with IBU across a 15-day period. Transcriptional dynamics were typical of an adaptive response with a peak of gene expression change at day 7 (177 features, representing about 11% of sequences available for analysis) and an almost full recovery at the end of the exposure period. Functional genomics by means of Gene Ontology term analysis unraveled typical mussel stress responses i.e. aminoglycan (chitin) metabolic processes but also more specific effects such as the regulation of NF-kappa B transcription factor activity. (C) 2016 Elsevier Ltd. All rights reserved.
- Contaminant effects in shore crabs (Carcinus maenas) from Ria Formosa LagoonPublication . Maria, Vera L.; Santos, M. A.; Bebianno, Maria JoĂŁoDefence and damage biomarkers signals were studied in female and male shore crabs Carcinus maenas transplanted between two sites at Ria Formosa Lagoon (South of Portugal). The cross transplantation occurred during 6 days at a hypothetical reference site (Ramalhete), and a contaminated site (OlhĂŁo). DNA unwinding technique was used for DNA integrity measurement. General enzymatic antioxidant responses in gills and hepatopancreas (catalase and gills glutathione peroxidase activities increase) showed that cross transplanted crabs (female and male) are suffering from pro-oxidant challenges at the OlhĂŁo site. Gills and hepatopancreas glutathione-S-transferase were reduced in both gender crabs transplanted from Ramalhete to OlhĂŁo. Metallothioneins induction occurred in crabs transplanted from Ramalhete to OlhĂŁo (contaminated by metals and polycyclic aromatic hydrocarbons (PAHs)). However metallothionein differences were also observed towards gender and organ specificities. High gills lipid peroxidation exist in male crabs transplanted from Ramalhete to OlhĂŁo, while in females it was the opposite. In both gender crabs from the OlhĂŁo site, gills DNA integrity decreased compared to the Ramalhete feral crabs. Moreover, hepatopancreas DNA integrity decreased in male crabs transplanted from OlhĂŁo to Ramalhete site which may be related to the environmental conditions (lower contamination levels) revealing the difficulty of selection of reference sites in field studies. Data demonstrated that female and male C. maenas antioxidant defences and damage biomarkers were sensitive to the mixture of contaminants present in these sites as well as good indicators of general stress.
- Contamination assessment of a coastal lagoon (Ria de Aveiro, Portugal) using defence and damage biochemical indicators in gill of Liza aurata â An integrated biomarker approachPublication . Oliveira, M.; Maria, Vera L.; Ahmad, I.; Serafim, Angela; Bebianno, Maria JoĂŁo; Pacheco, M.; Santos, M. A.Fish gill importance in toxicants uptake, bioconcentration and excretion allied to meagre knowledge on branchial damage/protection responses substantiate this study. Five critical sites in Ria de Aveiro (Portugal) were assessed in comparison with a reference site (Torreira), focusing on Liza aurata gill antioxidant defences versus damage (oxidative and genetic). Only in Barra fish displayed damage (lipid peroxidation) though no differences were found in antioxidants. In all other sites, except Rio, antioxidant alterations were found. Thus, fish from Gafanha, Laranjo and Vagos showed higher total glutathione, glutathione peroxidase and catalase. Higher glutathione reductase and glutathione S-transferase activity was also found in the first and the last sites, respectively. In Laranjo, metallothionein levels were higher though lower in Gafanha and Vagos. In general, damage was not accompanied by defences weakening confirming that predicting damage based on antioxidants depletion is not straightforward. The integrated biomarker response index ranked sites as: Gafanha>Barra>Laranjo>Vagos>Rio>Torreira.
- Evaluation of oxidative DNA lesions in plasma and nuclear abnormalities in erythrocytes of wild fish (Liza aurata) as an integrated approach to genotoxicity assessmentPublication . Oliveira, M.; Ahmad, I.; Maria, Vera L.; Ferreira, C. S. S.; Serafim, A.; Bebianno, Maria JoĂŁo; Pacheco, M.; Santos, M. A.Genetic lesions (8-hydroxy-2'-deoxyguanosine (8-OHdG) and erythrocytic nuclear abnormalities (ENA) were seasonally quantified in the blood of Liza aurata caught at Ria de Aveiro (Portugal), a multi-contaminated aquatic system. Thus, five critical sites were assessed and compared with a reference site (Torreira). Oxidative DNA damage was found in Gafanha (harbour-water area), Laranjo (metal-contaminated) and Vagos (contaminated with PAHs) in the spring; Rio Novo do Principe (near a former paper-mill effluent) in the autumn; Rio Novo do Principe and Vagos in the winter. ENA were higher than Torreira at VAG (spring and winter). Torreira did not display seasonal variation neither in terms of 8-OHdG or total ENA. A positive correlation between 8-OHdG and ENA was found, suggesting oxidative stress as a mechanism involved in the formation of ENA. This study clearly demonstrates the presence of DNA-damaging substances in Ria de Aveiro and recommends the use of 8-OHdG and ENA as biomarkers of environmental contamination.