Biomarkers in Solea Senegalensis Kaup, 1858 exposed to contaminated estuarine sediments: a multi-level approach

Thesis submitted to the Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia for the degree of Doctor of Philosophy in Environmental Sciences

Aeromonas spp. isolated from oysters (Crassostrea rhizophorea) from a natural oyster bed, Ceará, Brazil

Between April and October 2002, thirty fortnightly collections of oysters (Crassostrea rhizophorea) from a natural oyster bed at the Cocó River estuary in the Sabiaguaba region (Fortaleza, Ceará, Brazil) were carried out, aiming to isolate Aeromonas spp. strains. Oyster samples were submitted to the direct plating (DP) and the presence/absence (P/A) methods. Aeromonas were identified in 15 (50%) samples analyzed by the DP method and in 13 (43%) analyzed by the P/A method. A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii and Aeromonas sp. were isolated. The predominant species was A. veronii (both biovars), which was identified in 13 (43%) samples, followed by A. media in 11 (37%) and A. caviae in seven (23%). From the 59 strains identified, 28 (48%) presented resistance to at least one of the eight antibiotics tested.

N-acetyl-β -d-glucosaminidase activity in feral Carcinus maenasexposed to cadmium

Cadmium is a priority hazardous substance, persistent in the aquatic environment, with the capacity to interfere with crustacean moulting. Moulting is a vital process dictating crustacean growth, reproduction and metamorphosis. However, for many organisms, moult disruption is difficult to evaluate in the short term, what limits its inclusion in monitoring programmes. N-acetyl-β-d-glucosaminidase (NAGase) is an enzyme acting in the final steps of the endocrine-regulated moulting cascade, allowing for the cast off of the old exoskeleton, with potential interest as a biomarker of moult disruption. This study investigated responses to waterborne cadmium of NAGase activity of Carcinus maenas originating from estuaries with different histories of anthropogenic contamination: a low impacted and a moderately polluted one. Crabs from both sites were individually exposed for seven days to cadmium concentrations ranging from 1.3 to 2000 μg/L. At the end of the assays, NAGase activity was assessed in the epidermis and digestive gland. Detoxification, antioxidant, energy production, and oxidative stress biomarkers implicated in cadmium metabolism and tolerance were also assessed to better understand differential NAGase responses: activity of glutathione S-transferases (GST), glutathione peroxidase (GPx) glutathione reductase (GR), levels of total glutathiones (TG), lipid peroxidation (LPO), lactate dehydrogenase (LDH), and NADP+-dependent isocitrate dehydrogenase (IDH). Animals from the moderately polluted estuary had lower NAGase activity both in the epidermis and digestive gland than in the low impacted site. NAGase activity in the epidermis and digestive gland of C. maenas from both estuaries was sensitive to cadmium exposure suggesting its usefulness for inclusion in monitoring programmes. However, in the digestive gland NAGase inhibition was found in crabs from the less impacted site but not in those from the moderately contaminated one. Altered glutathione levels were observed in cadmium-treated crabs from the contaminated site possibly conferring enhanced tolerance to these animals through its chelator action. Investigation of enhanced tolerance should thus be accounted for in monitoring programmes employing NAGase as biomarker to avoid data misinterpretation.

Sertraline accumulation and effects in the estuarine decapod Carcinusmaenas: Importance of the history of exposure to chemical stress

Sertraline is widely prescribed worldwide and frequently detected in aquatic systems. There is, however, a remarkable gap of information on its potential impact on estuarine and coastal invertebrates. This study investigated sertraline accumulation and effects in Carcinus maenas. Crabs from a moderately contaminated (Lima) and a low-impacted (Minho) estuary were exposed to environmental and high levels of sertraline (0.05, 5, 500 μg L−1). A battery of biomarkers related to sertraline mode of action was employed to assess neurotransmission, energy metabolism, biotransformation and oxidative stress pathways. After a seven-day exposure, sertraline accumulation in crabs’ soft tissues was found in Lima (5 μg L−1: 15.3 ng L−1 ww; 500 μg L−1: 1010 ng L−1 ww) and Minho (500 μg L−1: 605 ng L−1 ww) animals. Lima crabs were also more sensitive to sertraline than those from Minho, exhibiting decreased acetylcholinesterase activity, indicative of ventilatory and locomotory dysfunction, inhibition of anti-oxidant enzymes and increased oxidative damage at ≥0.05 μg L−1. The Integrated Biomarker Response (IBR) index indicated their low health status. In addition, Minho crabs showed non-monotonic responses of acetylcholinesterase suggestive of hormesis. The results pointed an influence of the exposure history on differential sensitivity to sertraline and the need to perform evaluations with site-specific ecological receptors to increase relevance of risk estimations when extrapolating from laboratory to field conditions.

Integration of sediment contamination with multi–biomarker responses in a novel bioindicator candidate (Sepia officinalis) for risk assessment in impacted estuaries

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente

Numerical methods and tangible interfaces for pollutant dispersion simulation

Dissertação apresentada para obtenção do Grau de Doutor em Engenharia do Ambiente, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Can the complex mixture of sediment contaminants cause endocrine disruption on a benthic fish? A case study with Solea senegalensis

Pollution in coastal ecosystems is a serious threat to the biota and human populations there residing. Anthropogenic activities in these ecosystems are the main cause of contamination by endocrine disruption compounds (EDCs), which can interfere with hormonal regulation and cause adverse effects to growth, stress response and reproduction. Although the chemical nature of many EDCs is unknown, it is believed that most are organic contaminants. Under an environmental risk assessment for a contaminated estuary (the Sado, SW Portugal), the present work intended to detect endocrine disruption in a flatsfish, Solea senegalensis Kaup, 1858, and its potential relationship to organic toxicants. Animals were collected from two areas in the estuary with distinct influences (industrial and rural) and from an external reference area. To evaluate endocrine disruption, hepatic vitellogenin (VTG) concentrations in males and gonad histology were analysed. As biomarkers of exposure to organic contaminants, cytochrome P450 (CYP1A) induction and the ethoxyresorufin-O-deethylase (EROD) activity were determined. The results were contrasted to sediment contamination levels, which are overall considered low, although the area presents a complex mixture of toxicants. Either males or females were found sexually immature and showed no significant evidence of degenerative pathologies. However, hepatic VTG concentrations in males from the industrial area in estuary were superior than those from the Reference, even reaching levels comparable to those in females, which may indicate an oestrogenic effect resulting from the complex contaminant mixture. These individuals also presented higher levels of CYP1A induction and EROD activity, which is consistent with contamination by organic substances. The combination of the results suggest that the exposure of flatfish to an environment contaminated by mixed toxicants, even at low levels, may cause endocrine disruption, therefore affecting populations, which implies the need for further research in identification of potential EDCs, their sources and risks at ecosystem scale.

Ocorrência de Alevinos de Piramutaba (Brachyplatystoma vaillantii) na Baía de Marajó

Experimental fishing performed in the estuary of river Pará suggest that the bay of Marajó is a spawning and feeding ground for the Piramutaba, where differente age classes (20-600mm) are present during March-April.

Forest biomass and wood consumption In the lower course of the Amazon: a case study of rhe Urubueua Island

Várzea and terra-firme forests in the lower course of the Amazon were compared in terms of forest structure, wood volume increments and forest biomass. The wood volume of várzea forests was smaller than that of terra-firme forests, particularly when severe human intervention such as the cultivation of açaí palm occurred. The difference was even greater in the forest weight comparison because of the lower wood density of várzea trees. These trees are not directly influenced by water stress during the dry season, while late wood with a high density is formed in the terra-firme trees. The annual forest disappearance area due to firewood for tile factories was estimated to be about 276 ha on the island investigated, which had an area of 36,200 ha. Assuming that the forests are rotatively cultivated every 25 to 30 years, the total deforestation area is 6,870-6,948 ha in 25 years and 8,244~8,337 ha in 30 years. This result means that the balance between forest biomass and utilization is not in crisis, however, this balance might be lost as long as substitutive energy such as electricity is not supplied.

Time-variations of equivalent water heights'from Grace Mission and in-situ river stages in the Amazon basin

Gravity Recovery and Climate Experiment (GRACE) mission is dedicated to measuring temporal variations of the Earth's gravity field. In this study, the Stokes coefficients made available by Groupe de Recherche en Géodésie Spatiale (GRGS) at a 10-day interval were converted into equivalent water height (EWH) for a ~4-year period in the Amazon basin (from July-2002 to May-2006). The seasonal amplitudes of EWH signal are the largest on the surface of Earth and reach ~ 1250mm at that basin's center. Error budget represents ~130 mm of EWH, including formal errors on Stokes coefficient, leakage errors (12 ~ 21 mm) and spectrum truncation (10 ~ 15 mm). Comparison between in situ river level time series measured at 233 ground-based hydrometric stations (HS) in the Amazon basin and vertically-integrated EWH derived from GRACE is carried out in this paper. Although EWH and HS measure different water bodies, in most of the cases a high correlation (up to ~80%) is detected between the HS series and EWH series at the same site. This correlation allows adjusting linear relationships between in situ and GRACE-based series for the major tributaries of the Amazon river. The regression coefficients decrease from up to down stream along the rivers reaching the theoretical value 1 at the Amazon's mouth in the Atlantic Ocean. The variation of the regression coefficients versus the distance from estuary is analysed for the largest rivers in the basin. In a second step, a classification of the proportionality between in situ and GRACE time-series is proposed.

Chemical composition of phytoplankton from the estuaries of Eastern Amazonia

Phytoplankton is important bioindicator of chemical and biological modifications of natural ecosystems. The objective of this study was to determine the total chemical composition of the phytoplankton of the Pará and Mocajuba estuaries on the eastern coast of the Amazon region in the Brazilian state of Pará. The chemical composition of the surface water, bottom sediments (total sample and bioavailable fraction), and the phytoplankton were determined by inductively coupled plasma optical emission spectrometry. Phytoplankton contained high concentrations of Ca, P, Mn, Fe, Zn, Al, Ba, and Pb. The phytoplankton of the Mocajuba estuary is rich in Fe (2,967-84,750 µg g-1), while those from the Pará is rich in Al (1,216-15,389 µgg-1), probably reflecting divergent anthropogenic inputs. Both samples indicated a high bioconcentration factor derived from both the water and the bioavailable fraction, reflecting the efficiency of these organisms in the concentration of metals.

Relatório de atividade profissional

Dissertação de mestrado em Ciências - Formação Contínua de Professores (área de especialização em Biologia e Geologia)

Avaliação do transporte sedimentar na bacia do rio Lima baseada em ferramentas de modelação

Dissertação de mestrado integrado em Engenharia Civil

A celebração do contrato de seguro à distância

Dissertação de mestrado em Direito dos Contratos e da Empresa

საჯარო სკოლაში შიდა კონტროლის მექანიზმები და მართვის სტილი საქართველოში

საქართველოში განხორციელებული განათლების სისტემის რეფორმის ვექტორის ხშირმა ცვლამ გაზარდა სკოლების ცენტრალიზაციის პროცესი. ბოლო ორი წლის განმავლობაში ზოგადი განათლების შესახებ კანონში შევიდა ცვლილებები, რომლის შედეგად გაძლიერდა განათლებისა და მეცნიერების სამინისტროს როლი სკოლის მართვის პროცესში. მინისტრის მიერ გამოცემულმა ბრძანებებმა , დირექტორის სტანდარტის, მასწავლებლისა და მოსწავლის ეთიკის სტანდარტის დამტკიცებამ , 2010 წლის აგვისტო- სექტემბერში საქართველოს საჯარო სკოლებმა განაახლეს სკოლის შინაგანაწესი. სკოლის შინაგანაწესებმა ასახა სისტემის მართვაში შესული ცვლილებები. რამაც საბოლოო ჯამში ნულოვანი ტოლერანტობის ტენდენციების გაძლიერებას შეუწყო ხელი. სკოლის დისციპლინური მართვის პროცედურები გაძლიერდა, სკოლის შინაგანაწესებმა განსაზღვრა ხისტი სადამსჯელო ზომები დისციპლინურ გადაცდომებთან დაკავშირებით, თუმცა დისციპლინური გადაცდომების ზუსტი აღწერა შემუშავებულ შინაგანაწესებში არ ჩანს. ამ ნაშრომში შესწავლილია საქართველოს საჯარო სკოლის შინაგანაწესები, რომლითაც განისაზღვრება სკოლის მართვის სტილი და კონტროლის მექანიზმები. ნაშრომში ნაჩვენებია განათლების პოლიტიკის სიტემური, კერძოდ დისციპლინური მართვის სისტემის რეფორმირების საჭიროება.