Integrated biomarker responses of an estuarine invertebrate to high abiotic stress and decreased metal contamination

An integrated chemical-biological effects monitoring was performed in 2010 and 2012 in two NW Iberian estuaries under different anthropogenic pressure. One is low impacted and the other is contaminated by metals. The aim was to verify the usefulness of a multibiomarker approach, using Carcinus maenas as bioindicator species, to reflect diminishing environmental contamination and improved health status under abiotic variation. Sampling sites were assessed for metal levels in sediments and C. maenas, water abiotic factors and biomarkers (neurotoxicity, energy metabolism, biotransformation, anti-oxidant defences, oxidative damage). High inter-annual and seasonal abiotic variation was observed. Metal levels in sediments and crab tissues were markedly higher in 2010 than in 2012 in the contaminated estuary. Biomarkers indicated differences between the study sites and seasons and an improvement of effects measured in C. maenas from the polluted estuary in 2012. Integrated Biomarker Response (IBR) index depicted sites with higher stress levels whereas Principal Component Analysis (PCA) showed associations between biomarker responses and environmental variables. The multibiomarker approach and integrated assessments proved to be useful to the early diagnosis of remediation measures in impacted sites.

Fresh-cut aromatic herbs: Nutritional quality stability during shelf-life

Fresh-cut vegetables are a successful convenient healthy food. Nowadays, the presence of new varieties of minimally processed vegetables in the market is common in response to the consumers demand for new flavours and high quality products. Within the most recent fresh-cut products are the aromatic herbs. In this work, the objective was to evaluate the nutritional quality and stability of four fresh-cut aromatic herbs. Several physicochemical quality characteristics (colour, pH, total soluble solids, and total titratable acidity) were monitored in fresh-cut chives, coriander, spearmint and parsley leaves, stored under refrigeration (3 ± 1 ºC) during 10 days. Their nutritional composition was determined, including mineral composition (phosphorous, potassium, sodium, calcium, magnesium, iron, zinc, manganese and copper) and fat- and water-soluble vitamin contents. Total soluble phenolics, flavonoids and the antioxidant capacity were determined by spectrophotometric methods. The aromatic herbs kept their fresh appearance during the storage, maintaining their colour throughout shelf-life. Their macronutrient composition and mineral content were stable during storage. Coriander had the highest mineral and fatsoluble vitamin content, while spearmint showed the best scores in the phenolic, flavonoid and antioxidant capacity assays. Vitamins and antioxidant capacity showed some variation during storage, with a differential behaviour of each compound according to the sample.

Joint effects of salinity and the antidepressant sertraline on the estuarine decapod Carcinus maenas

Concurrent exposure of estuarine organisms to man-made and natural stressors has become a common occurrence. Numerous interactions of multiple stressors causing synergistic or antagonistic effects have been described. However, limited information is available on combined effects of emerging pharmaceuticals and natural stressors. This study investigated the joint effects of the antidepressant sertraline and salinity on Carcinus maenas. To improve knowledge about interactive effects and potential vulnerability,experiments were performed with organisms from two estuaries with differing histories of exposure to environmental contamination. Biomarkers related to mode of action of sertraline were employed to assess effects of environmentally realistic concentrations of sertraline at two salinity levels. Synergism and antagonism were identified for biomarkers of cholinergic neurotransmission, energy production,anti-oxidant defences and oxidative damage. Different interactions were found for the two study sites highlighting the need to account for differences in tolerance of local ecological receptors in risk evaluations.

Targeting specific nutrient deficiencies in proteinrestricted diets: some practical facts in PKU dietary management

Among aminoacidopathies, phenylketonuria (PKU) is the most prevalent one. Early diagnosis in the neonatal period with a prompt nutritional therapy (low natural-protein and phenylalanine diet, supplemented with phenylalanine-free amino acid mixtures and special low-protein foods) remains the mainstay of the treatment. Data considering nutrient contents of cooked dishes is lacking. In this study, fourteen dishes specifically prepared for PKU individuals were analysed, regarding the lipid profile and iron and zinc contents. These dishes are poor sources of essential nutrients like Fe, Zn or n-3 fatty acids, reinforcing the need for adequate supplementation to cover individual patients’ needs. This study can contribute to a more accurate adjustment of PKU diets and supplementation in order to prevent eventual nutritional deficiencies. This study contributes to a better understanding of nutrient intake from PKU patients’ meals, showing the need for dietary supplementation.

Teor mineral e capacidade antioxidante em chás e infusões

Mestrado em Engenharia Química – Ramo Optimização Energética na Indústria Química

Lettuce (Lactuca sativa L.) leaf-proteome profiles after exposure to cylindrospermopsin and a microcystin-LR/cylindrospermopsin mixture: A concentration-dependent response

The intensification of agricultural productivity is an important challenge worldwide. However, environmental stressors can provide challenges to this intensification. The progressive occurrence of the cyanotoxins cylindrospermopsin (CYN) and microcystin-LR (MC-LR) as a potential consequence of eutrophication and climate change is of increasing concern in the agricultural sector because it has been reported that these cyanotoxins exert harmful effects in crop plants. A proteomic-based approach has been shown to be a suitable tool for the detection and identification of the primary responses of organisms exposed to cyanotoxins. The aim of this study was to compare the leaf-proteome profiles of lettuce plants exposed to environmentally relevant concentrations of CYN and a MC-LR/CYN mixture. Lettuce plants were exposed to 1, 10, and 100 lg/l CYN and a MC-LR/CYN mixture for five days. The proteins of lettuce leaves were separated by twodimensional electrophoresis (2-DE), and those that were differentially abundant were then identified by matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF/TOF MS). The biological functions of the proteins that were most represented in both experiments were photosynthesis and carbon metabolism and stress/defense response. Proteins involved in protein synthesis and signal transduction were also highly observed in the MC-LR/CYN experiment. Although distinct protein abundance patterns were observed in both experiments, the effects appear to be concentration-dependent, and the effects of the mixture were clearly stronger than those of CYN alone. The obtained results highlight the putative tolerance of lettuce to CYN at concentrations up to 100 lg/l. Furthermore, the combination of CYN with MC-LR at low concentrations (1 lg/l) stimulated a significant increase in the fresh weight (fr. wt) of lettuce leaves and at the proteomic level resulted in the increase in abundance of a high number of proteins. In contrast, many proteins exhibited a decrease in abundance or were absent in the gels of the simultaneous exposure to 10 and 100 lg/l MC-LR/CYN. In the latter, also a significant decrease in the fr. wt of lettuce leaves was obtained. These findings provide important insights into the molecular mechanisms of the lettuce response to CYN and MC-LR/CYN and may contribute to the identification of potential protein markers of exposure and proteins that may confer tolerance to CYN and MC-LR/CYN. Furthermore, because lettuce is an important crop worldwide, this study may improve our understanding of the potential impact of these cyanotoxins on its quality traits (e.g., presence of allergenic proteins).

Picocyanobacteria From a Clade Of Marine Cyanobium Revealed Bioactive Potential Against Microalgae, Bacteria, And Marine

The production of bioactive compounds either toxic or with pharmacological applications by cyanobacteria is well established. However, picoplanktonic forms within this group of organisms have rarely been studied in this context. In this study, the toxicological potential of picocyanobacteria from a clade of marine Cyanobium strains isolated from the Portuguese coast was examined using different biological models. First, strains were identified by applying morphological and molecular approaches and cultured under lab conditions. A crude extract and three fractions reflecting a preliminary segregation of lipophilic metabolites were tested for toxicity with the marine microalga Nannochloropsis sp., the bacteria Pseudomonas sp., the brine shrimp Artemia salina, and fertilized eggs of the sea urchin Paracentrotus lividus. No significant apparent adverse effects were noted against Artemia salina. However, significant adverse effects were found in all other assays, with an inhibition of Nannochloropsis sp. and Pseudomonas sp. growth and marked reduction in Paracentrotus lividus larvae length. The results obtained indicated that Cyanobium genus may serve as a potential source of interesting bioactive compounds and emphasize the importance of also studying smaller picoplanktonic fractions of marine cyanobacteria.

The role of lifelong learning (LLL) in the creation of a european knowledge- based society

This chapter appears in Encyclopaedia of Human Resources Information Systems: Challenges in e-HRM edited by Torres-Coronas, T. and Arias-Oliva, M. Copyright 2009, IGI Global, www.igi-global.com. Posted by permission of the publisher. URL:http://www.igi-pub.com/reference/details.asp?id=7737

Desenvolvimento de um método potenciométrico para deteção de salmonella typhimurium

A Salmonella é um microrganismo responsável por grande parte das doenças alimentares, podendo por em causa a saúde pública da área contaminada. Uma deteção rápida, eficiente e altamente sensível e extremamente importante, sendo um campo em franco desenvolvimento e alvo de variados e múltiplos estudos na comunidade cientifica atual. Foi desenvolvido um método potenciométrico para a deteção de Salmonellas, com elétrodos seletivos de iões, construídos em laboratório com pontas de micropipetas, fios de prata e sensores com composição otimizada. O elétrodo indicador escolhido foi um ESI seletivo a cadmio, para redução da probabilidade de interferências no método, devido a pouca abundancia do cadmio em amostras alimentares. Elétrodos seletivos a sódio, elétrodos de Ag/AgCl de simples e de dupla juncão foram também construídos e caracterizados para serem aplicados como elétrodos de referência. Adicionalmente otimizaram-se as condições operacionais para a analise potenciométrica, nomeadamente o elétrodo de referencia utilizado, condicionamento dos elétrodos, efeito do pH e volume da solução amostra. A capacidade de realizar leituras em volumes muito pequenos com limites de deteção na ordem dos micromolares por parte dos ESI de membrana polimérica, foi integrada num ensaio com um formato nao competitivo ELISA tipo sanduiche, utilizando um anticorpo primário ligado a nanopartículas de Fe@Au, permitindo a separação dos complexos anticorpo-antigénio formados dos restantes componentes em cada etapa do ensaio, pela simples aplicação de um campo magnético. O anticorpo secundário foi marcado com nanocristais de CdS, que são bastante estáveis e é fácil a transformação em Cd2+ livre, permitindo a leitura potenciométrica. Foram testadas várias concentrações de peroxido de hidrogénio e o efeito da luz para otimizar a dissolução de CdS. O método desenvolvido permitiu traçar curvas de calibração com soluções de Salmonellas incubadas em PBS (pH 4,4) em que o limite de deteção foi de 1100 CFU/mL e de 20 CFU/mL, utilizando volumes de amostra de 10 ƒÊL e 100 ƒÊL, respetivamente para o intervalo de linearidade de 10 a 108 CFU/mL. O método foi aplicado a uma amostra de leite bovino. A taxa de recuperação media obtida foi de 93,7% } 2,8 (media } desvio padrão), tendo em conta dois ensaios de recuperação efetuados (com duas replicas cada), utilizando um volume de amostra de 100 ƒÊL e concentrações de 100 e 1000 CFU/mL de Salmonella incubada.

Diagnosis of cytomegalovirus infections by qualitative and quantitative PCR in HIV infected patients

A high incidence of cytomegalovirus (CMV) infections is observed in Brazil. These viruses are causatives of significant morbidity and mortality among patients with advanced human immunodeficiency virus (HIV) infection. This work, shows the application of a PCR on determination of CMV load in the buffy coat and plasma. We analyzed the samples of 247 HIV infected patients in order to diagnose CMV infection and disease. We developed a semi-quantitative PCR that amplifies part of the glycoprotein B (gB) gene of CMV. The semi-quantitative PCR was carried out only in positive clinical samples in a qualitative PCR confirmed by a nested-PCR. CD4 lymphocyte count, HIV viral load and CMV disease symptom were correlated with CMV load. CMV genome was detected in the buffy coat of 82 of 237 (34.6%) patients, in 10 of these the CMV load was determined varying between 928 and 332 880 viral copies/mug DNA. None of these 237 patients developed any suggestive manifestation of CMV disease. For the other 10 HIV infected patients selected based on the suspicion of CMV disease, CMV genome was detected in only one case. This patient presented a high CMV load, 8 000 000 copies/mug DNA, and developed a disseminated form of CMV disease including hepatitis and retinitis. Our results were greatly influenced by the impact of the highly active antiretroviral therapy that reduced incidence of CMV viremia and occurrence of CMV disease in the HIV infected patients.

Fronteira e guerra civil de Espanha. Dominação, resistência e usos da memória em Barrancos

Tese de Doutoramento em Antropologia especialidade em Poder, Resistência e Movimentos Sociais

Botulism: a laboratory investigation on biological and food samples from cases and outbreaks in Brazil (1982-2001)

Laboratory investigation of botulism from 1982 to 2001 confirmed the occurrence of eight positive outbreaks/cases of botulism in Brazil. From those, type A botulism was observed in seven of them. Biological material of one case (serum and feces) was positive in the first step of the bioassay, but the amount of sample was not sufficient for typification. One of the outbreaks that occurred in 2001 was negative for botulinum toxin in samples of serum, gastric washing and feces, collected eight days before the onset of the symptoms in the affected person who was clinically diagnosed as presenting the disease. Other two cases presenting compatible clinical diagnoses presented negative results. However, in those cases, the collection of samples was (1) after antiserum administration or (2) later than eight days of the onset of symptoms. Investigation was performed by mouse bioassay, as described in the Compendium of Methods for the Microbiological Examination of Foods (compiled by American Public Health Association - APHA)11, using specific antiserum from Centers for Disease Control (CDC), USA.

Avaliação e Otimização do Consumo de Água no Processo e do Sistema de Tratamento de Efluentes

O elevado consumo de água associado à escassez deste recurso contribuiu para que alternativas de reutilização/reciclagem de água fossem estudadas que permitam diminuir o seu consumo e minimizar a dependência das indústrias. Monitorizar e avaliar os consumos de água, a nível industrial, é imprescindível para assegurar uma gestão sustentável dos recursos hídricos, sendo este o objetivo da presente dissertação. As alternativas encontradas na unidade industrial em estudo foram a substituição do equipamento sanitário e o aproveitamento do efluente tratado para operações de lavagem e/ou arrefecimento por contacto direto. A maioria do equipamento sanitário não é eficiente, tendo-se proposto a substituição desse sistema por um de menor consumo que permitirá uma poupança de 30 % no consumo de água, que corresponderá a 12 149,37 €/ano, sendo o retorno do investimento estimado em 3 meses. O efluente industrial na entrada da ETAR e nas diferentes etapas - tratamento primário de coagulação/floculação; tratamento secundário ou biológico em SBR; tratamento terciário de coagulação/floculação - foi caracterizado através da medição da temperatura, pH, oxigénio dissolvido e pela determinação da cor, turvação, sólidos suspensos totais (SST), azoto total, carência química de oxigénio (CQO), Carência Bioquímica de Oxigénio ao fim de 5 dias (CBO5) e razão CBO5/CQO. Esta caracterização permitiu avaliar o efluente industrial bruto que se caracteriza por um pH alcalino (8,3 ± 1,7); condutividade baixa (451 ± 200,2 μS/cm); elevada turvação (11 255 ± 8812,8 FTU); cor aparente (63 670 ± 42293,4 PtCo) e cor verdadeira (33 621 ± 19547,9 PtCo) elevadas; teores elevados de CQO (24 753 ± 11806,7 mg/L O2) SST (5 164 ± 3845,5 mg/L) e azoto total (718 mg/L) e um índice de biodegradabilidade baixo (razão CBO5/CQO de 1,4). Este estudo permitiu verificar que a eficiência global do tratamento do efluente foi 82 % na remoção da turvação, 83 % na remoção da cor aparente, 96 % na remoção da cor verdadeira, 85 % na remoção da CQO e 30 % na remoção dos SST. Quanto às eficiências de remoção associadas ao tratamento primário no que diz respeito à turvação, cor aparente, CQO e SST, apresentam valores inferiores aos referidos na literatura para o mesmo tipo de tratamento em efluentes similares. As eficiências de remoção obtidas no tratamento secundário são inferiores às do tratamento primário: turvação, cor aparente, CQO e SST, pelo que procurou-se otimizar a primeira etapa do processo de tratamento Neste estudo de otimização estudou-se a influência de cinco coagulantes – Sulfato de Alumínio, PAX XL – 10, PAX 18, cloreto de ferro e a conjugação de PAX 18 com sulfato de ferro - e seis floculantes – Superfloc A 150, Superfloc A 130, PA 1020, Ambifloc 560, Ambifloc C58 e Rifloc 54 - no tratamento físico-químico do efluente. O PAX 18 e o Ambifloc 560 UUJ foram os que apresentaram as mais elevadas eficiências de remoção (99,85 % na cor, 99,87 % na turvação, 90,12 % na CQO e 99,87 % nos SST). O custo associado a este tratamento é de 1,03 €/m3. Pela comparação com os critérios de qualidade no guia técnico ERSAR, apenas o parâmetro da CQO excede o valor, contudo o valor obtido permite diminuir os custos associados a um tratamento posterior para remoção da CQO remanescente no efluente residual tratado.

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.

A potentiometric magnetic immunoassay for rapid detection of Salmonella typhimurium

Potentiometric detection with homemade polymeric membrane microelectrodes was coupled to a magnetic sandwich immunoassay for Salmonella typhimurium determination. Cadmium and sodium ion selective electrodes were used respectively as indicator and pseudo-reference electrodes and were prepared in pipette tips to allow potentiometric measurements in microliter sample volumes. In the proposed method, the concentration of S. typhimurium was proportional to the amount of cadmium released upon dissolution of a CdS nanoparticle labeled to the secondary detection antibody. The limit of detection was 2 cells per 100 μL. The immunomagnetic assay with potentiometric detection is suitable for sensitive and rapid (average total time per assay of 75 minutes) detection of S. typhimurium in milk samples. The proposed method is easy to perform, safe, sensitive, and low cost and has potential for in situ analysis.