Development of polyaniline microarray electrodes for cadmium analysis

Disposable screen-printed electrodes (SPCE) were modified using a cosmetic product to partially block the electrode surface in order to obtain a microelectrode array. The microarrays formed were electropolymerized with aniline. Scanning electron microscopy was used to evaluate the modified and polymerized electrode surface. Electrochemical characteristics of the constructed sensor for cadmium analysis were evaluated by cyclic and square-wave voltammetry. Optimized stripping procedure in which the preconcentration of cadmium was achieved by depositing at –1.20 V (vs. Ag/AgCl) resulted in a well defined anodic peak at approximately –0.7 V at pH 4.6. The achieved limit of detection was 4 × 10−9 mol dm−3. Spray modified and polymerized microarray electrodes were successfully applied to quantify cadmium in fish sample digests

Environmental Particulate Matter Levels during 2017 Large Forest Fires and Megafires in the Center Region of Portugal: A Public Health Concern?

This work characterizes the dimension and the exceptionality of 2017 large- and mega-fires that occurred in the center region of Portugal through the assessment of their impact on the ambient levels of particulate matter (PM10 and PM2.5), retrieved from local monitoring stations, and the associated public health risks. PM10 and PM2.5 concentrations were increased during the occurrence of large fires and megafires, with daily concentrations exceeding the European/national guidelines in 7–14 and 1–12 days of 2017 (up to 704 µg/m3 for PM10 and 46 µg/m3 for PM2.5), respectively. PM10 concentrations were correlated with total burned area (0.500 0.05) and with monthly total burned area/distance2 (0.500 0.05). The forest fires of 2017 took the life of 112 citizens. A total of 474 cases of hospital admissions due to cardiovascular diseases and 3524 cases of asthma incidence symptoms per 100,000 individuals at risk were assessed due to exposure to 2017 forest fires. Real-time and in situ PM methodologies should be combined with protection action plans to reduce public health risks. Portuguese rural stations should monitor other health-relevant pollutants (e.g., carbon monoxide and volatile organic compounds) released from wildfires to allow performing more robust and comprehensive measurements that will allow a better assessment of the potential health risks for the exposed populations.This work was financially supported by European Union (FEDER funds through COMPETE) and National Funds (Fundação para a Ciência e Tecnologia) [projects UIDB/50006/2020, UID/EQU/00511/2013-LEPABE], by the FCT/MEC with national funds and cofounded by FEDER in the scope of the P2020 Partnership Agreement. This study was also supported by the project “PCIF/SSO/0017/2018- A panel of (bio)markers for the surveillance of firefighter’s health and safety”, funded by Portuguese National Funds through FCT—Fundação para a Ciência e Tecnologia. M. Oliveira thanks to FCT/MCTES for the CEEC-Individual 2017 Program Contract [CEECIND/03666/2017].info:eu-repo/semantics/publishedVersio

Resistência a antibióticos de último recurso em ambientes naturais

Doutoramento em BiologiaLast-resort antibiotics are the final line of action for treating serious infections caused by multiresistant strains. Over the years the prevalence of resistant bacteria has been increasing. Natural environments are reservoirs of antibiotic resistance, highly influenced by human-driven activities. The importance of aquatic systems on the evolution of antibiotic resistance is highlighted from the assumption that clinically-relevant resistance genes have originated in strains ubiquitous in these environments. We hypothesize that: a) rivers are reservoirs and disseminators of antibiotic resistance; b) anthropogenic activities potentiate dissemination of resistance to last-resort antibiotics. Hence, the main goal of the work is to compare the last-resort antibiotics resistome, in polluted and unpolluted water. Rivers from the Vouga basin, exposed to different anthropogenic impacts, were sampled. Water quality parameters were determined to classify rivers as unpolluted or polluted. Two bacterial collections were established enclosing bacteria resistant to cefotaxime (3rd generation cephalosporin) and to imipenem (carbapenem). Each collection was characterized regarding: phylogenetic diversity, antibiotic susceptibility, resistance mechanisms and mobile genetic elements. The prevalence of cefotaxime- and imipenem-resistant bacteria was higher in polluted water. Results suggested an important role in the dissemination of antibiotic resistance for Enterobacteriaceae, Pseudomonas and Aeromonas. The occurrence of bacteria resistant to non-beta-lactams was higher among isolates from polluted water as also the number of multiresistant strains. Among strains resistant to cefotaxime, extended-spectrum beta-lactamase (ESBL) genes were detected (predominantly blaCTX-M-like) associated to mobile genetic elements previously described in clinical strains. ESBL-producers were often multiresistant as a result of co-selection mechanisms. Culture-independent methods showed clear differences between blaCTX-M-like sequences found in unpolluted water (similar to ancestral genes) and polluted water (sequences identical to those reported in clinical settings). Carbapenem resistance was mostly related to the presence of intrinsically resistant bacteria. Yet, relevant carbapenemase genes were detected as blaOXA-48-like in Shewanella spp. (the putative origin of these genes), and blaVIM-2 in Pseudomonas spp. isolated from polluted rivers. Culture-independent methods showed an higher than the previously reported diversity of blaOXA-48-like genes in rivers. Overall, clear differences between polluted and unpolluted systems were observed, regarding prevalence, phylogenetic diversity and susceptibility profiles of resistant bacteria and occurrence of clinically relevant antibiotic resistance genes, thus validating our hypotheses. In this way, rivers act as disseminators of resistance genes, and anthropogenic activities potentiate horizontal gene transfer and promote the constitution of genetic platforms that combine several resistance determinants, leading to multiresistance phenotypes that may persist even in the absence of antibiotics.Antibióticos de último recurso são usados no tratamento de infecções graves causadas por estirpes multiresistentes. A prevalência de bactérias resistentes a estes antibióticos tem aumentado. Os ambientes naturais, influenciados pela actividade humana, são reservatórios de bactérias resistentes e de genes de resistência. Vários genes de resistência com grande impacto na clínica têm presumivelmente origem em estirpes ubíquas em sistemas aquáticos, o que realça a importância destes ambientes na evolução de resistência. Este estudo assenta nas seguintes hipóteses: a) os rios são reservatórios e disseminadores de resistência a antibióticos; b) as atividades antropogénicas potenciam a disseminação de resistência a antibióticos de último recurso nestes ambientes. Assim, foi estabelecido como objectivo comparar o resistoma ambiental referente a antibióticos de último recurso, em rios poluídos e não poluídos. Foram amostrados rios na Bacia Hidrográfica do Vouga, expostos a diferentes impactos antropogénicos. Os rios foram classificados como poluídos e não poluídos de acordo com parâmetros de qualidade da água. Duas colecções foram estabelecidas: bactérias resistentes a cefotaxima (cefalosporina de 3ª geração) e a imipenemo (carbapenemo). Cada colecção foi caracterizada em termos de diversidade filogenética, susceptibilidade a antibióticos, mecanismos de resistência e elementos genéticos móveis. A prevalência de bactérias resistentes foi superior em águas poluídas. Os resultados sugerem que nestes ambientes Enterobacteriaceae, Pseudomonas e Aeromonas têm um papel importante na disseminação de resistência. Os níveis de resistência a não beta-lactâmicos foram superiores em águas poluídas, assim como o número de estirpes multiresistentes. Detectaram-se genes de beta-lactamases de espectro alargado, associados a elementos genéticos móveis previamente descritos em isolados clínicos. Métodos independentes do cultivo revelaram diferenças claras entre a diversidade de sequências do tipo blaCTX-M em rios poluídos (idênticas às encontradas em isolados clínicos) e não poluídos (similares a genes ancestrais). A resistência a carbapenemos foi maioritariamente relacionada com a presença de bactérias intrinsecamente resistentes. No entanto, foram identificados genes de carbapenemases relevantes tais como blaOXA-48 em Shewanella spp. (origem putativa destes genes) e blaVIM-2 em Pseudomonas spp. de rios poluídos. Métodos independentes do cultivo mostraram que, nestes rios, a diversidade de genes similares a blaOXA-48 é superior ao que tem sido relatado. Detectaramse diferenças evidentes entre rios poluídos e não poluídos, em termos de prevalência, diversidade filogenética e susceptibilidade a antibióticos em bactérias resistentes e ocorrência de genes de resistência clinicamente relevantes. Estes dados validam as hipóteses colocadas. Assim, estes sistemas aquáticos actuam como reservatórios de genes de resistência. As actividades antropogénicas potenciam a disseminação destes genes e a constituição de plataformas genéticas complexas, originando fenótipos de multiresistência que poderão persistir mesmo na ausência de antibióticos

Caracterização molecular de estirpes de Aeromonas spp. resistentes a antibióticos beta-lactâmicos

Mestrado em Microbiologia MolecularEstudos feitos em todo o mundo têm demonstrado que Aeromonas spp. estão distribuídas universalmente e podem ser isoladas de amostras clínicas, alimentares e ambientais. São exemplo de microrganismos patogénicos emergentes. Em sistemas de aquacultura, as infecções bacterianas são a causa principal das quebras de produtividade. O uso intensivo de antibióticos beta-lactâmicos como agentes terapêuticos levou ao aparecimento de estirpes multiresistentes. Neste trabalho, foram analisadas estirpes de Aeromonas isoladas de amostras de tecido da pele e rins de trutas arco-íris (Oncorhynchus mykiss) de uma aquacultura da Universidade de Trás-os-Montes e Alto Douro. Foram incluídas também estirpes de referência. Os testes de susceptibilidade a antibióticos beta-lactâmicos revelaram que a maior parte dos isolados é resistente a penicilinas e cefalosporinas de 1ª geração. Em 7 isolados foi detectada resistência a carbapenemos (imipenemo). Por forma a caracterizar o mecanismo de resistência dos isolados resistentes ao imipenemo, foram desenhados "primers" para amplificar por PCR genes de resistência homólogos a cphA e/ou imiS, que codificam as metalo-beta-lactamases CphA e ImiS, já descritas em Aeromonas spp.. Em 5 dos isolados e em 2 estirpes referência, foi possível amplificar por PCR, com os "primers" desenhados, Aer- R e Aer-F, um fragmento com o tamanho esperado, cerca de 670 pb, a partir do DNA total das estirpes de Aeromonas spp.. A análise das sequências nucleotídicas e das sequências de aminoácidos deduzidas revelou tratarem-se de fragmentos de DNA de genes homólogos a cphA/imiS, que codificam para metalo-beta-lactamases de classe B, subclasse B2. Procedeu-se também à detecção de sequências homólogas por hibridação DNA-DNA, usando uma sonda marcada por PCR com os "primers" Aer-R e Aer-F. Ocorreu hibridação entre o fragmento de DNA usado como sonda e o DNA de todos os isolados que apresentavam actividade de carbapenemase. Pretendeu-se, também, caracterizar os isolados ao nível molecular, através de técnicas de tipagem genética como ARDRA e rep-PCR. Os perfis ARDRA obtidos por digestão com MboI e AluI permitiram identificar os isolados como Aeromonas bestiarum Aeromonas hydrophila e Aeromonas media. Os perfis REP- e BOX-PCR resultantes mostraram-se mais complexos que os perfis ARDRA e permitiram avaliar o grau de diversidade genómica dos isolados, mostrando-se úteis na tipagem genética de isolados deste grupo de procariotas.Worldwide studies have demonstrated that Aeromonas spp. are universally distributed and widely isolated from clinical, environmental and food samples. They are an example of emerging bacterial pathogens. In aquaculture explorations, bacterial infectious diseases are the major cause of decreases in productivity. The extensive use of beta-lactam antibiotics as therapeutic agents has generated multiresistant strains. In this work, tissue samples from the skin and kidney of rainbow trouts (Oncorhynchus mykiss) from a fish farm at the Universidade de Trás-os-Montes e Alto Douro were examined for the presence of Aeromonas spp. Reference strains were also included in this study. The susceptibility to antibiotics tests revealed that most of these strains are resistant to penicillins and 1st generation cephalosporins. It was detected resistance to carbapenems (imipenem) in 7 aeromonads. In order to understand the resistance mechanism of the strains that showed resistance to carbapenems, primers Aer-R and Aer-F were designed to detect by PCR, homologous genes to cphA/imiS, which code for the metallo-beta-lactamases CphA and ImiS, already described in Aeromonas spp. In 5 isolates and in 2 reference strains, it was possible to amplify by PCR a DNA fragment with 670 bp as expected, whose sequence was determined. The nucleotide and deduced amino acid sequence analysis revealed that these are DNA fragments homologous to cphA/imiS, that code for class B, subclass B2 metallo-betalactamases. DNA-DNA hybridization was also applied using a DNA fragment, amplified by PCR with primers Aer-R and Aer-F, as probe. There was hybridization between the probe and the DNA from all the strains that showed resistance to carbapenems. Aeromonads isolates were also compared using different molecular typing techniques like ARDRA and rep-PCR. ARDRA profiles obtained from enzymatic digestion of rDNA 16S wilth MboI and AluI, allowed the identification of strains as Aeromonas bestiarum, Aeromonas hydrophila and Aeromonas media. Fingerprinting by REP- and BOX- PCR provided complex genomic profiles, proving that these are good methods for typing aeromonads

Caracterização e análise de custos da Disfagia Orafaríngea numa Unidade de Cuidados Continuados de média Duração e Reabilitação

Na saúde, importa responder a necessidades ilimitadas gerindo recursos limitados. Para isso, a Economia da Saúde inclui análises económicas de custos e consequências que estruturam as decisões dos gestores. A Disfagia Orofaríngea (DO) é uma dificuldade na deglutição que afeta a movimentação do bolo alimentar da cavidade oral até ao esófago. É prevalente em múltiplas condições clínicas, provoca malnutrição/desidratação, diminuição da qualidade de vida, Pneumonia por Aspiração (PA), agravamento clínico, morte e custos. O objetivo desta dissertação é analisar a prevalência, caracterização sociodemográfica e clínica, assim como descrever p acréscimo de custos de doentes com DO numa Unidade de Cuidados Continuados de Média Duração e Reabilitação (UCCMDR). Para cumprir os objetivos, realizou-se um estudo de caso descritivo, quantitativo, retrospetivo, transversal e uma análise de descrição do custo da doença. Constatou-se numa prevalência de 43.8% de doentes com DO, com média de idades de 76.2 (DP = 11.2) e baixa escolaridade. 56.14% eram do género masculino, 59.64% tinham Acidente Vascular Cerebral (AVC), 15.79% tiveram PA, 50.87% apresentavam malnutrição, 43.85% necessitavam de sonda Nasogástrica (SNG) e 85.96% tinham origem hospitalar. Identificou-se um aumento de 17.81 dias de internamento e acréscimo monetário total de 87 954,55€. Este estudo evidencia o impacto da DO nas UCCMDR, enriquecendo os estudos nesta área em Portugal

As cinco leis para a alfabetização midiática e informacional e sua relação com os fatores críticos de sucesso no âmbito da gestão do conhecimento.

Objective. The objective of this work is to understand the relationship between the Critical Success Factors and the 5 Laws of Information Media Literacy, in order to observe if it is possible based on such factors to develop information literacy on organizational subjects. Design/Methodology/Approach. The identification of Critical Success Factors constitutes an important phase of knowledge management in organizations, and their analysis is a key element for the success of the implementation of knowledge management. The Information Literacy, in its turn, seeks to develop skills and abilities related to the conscious use of information. The research is a documentary study, exploratory and qualitative, followed by a contrastive analysis between the Critical Success Factors identified in the literature studied and the Five Laws for Informational Literacy. Results/Discussion. It was possible to compose a systematizing table containing the main Critical Success Factors found and, thus, to analyze the main bases that impact said factors, as well as to identify the interrelationships from the aspects presented by the Five Laws. Conclusions.Although the Critical Success Factors are (and should be) adaptable to different organizational realities, they permeate aspects seeking harmony between people, technology and organizational structure. From this perspective, it is possible to improve the organizational conditions and the implementation of knowledge management programs and/or projects anchored in the main pillars identified. Originality/Value. UNESCO incorporated Media and Information Literacy as part of its mission in 2017, stressing that the development of skills in these two areas provides the knowledge, skills and attitudes required for life and work in the 21st Century. In bringing these laws for discussion in the Information Science field we show support for Unesco’s positioning, as well as reinforcing the need for the area’s activity within the knowledge management

Comparison of chickpea rhizobia isolates from diverse Portuguese natural populations based on symbiotic effectiveness and DNA fingerprint

Abstract: Aims: To test the hypothesis that differences in chickpea yields obtained in four distinct Portuguese regions (Beja, Elvas-Casas Velhas , Elvas-Estacao Nacional de Melhoramento de Plantas (ENMP) and Evora) could be due to variation between the natural rhizobia populations. Methods and Results: Estimation of the size of the different rhizobial populations showed that Elvas-ENMP population was the largest one. Elvas-ENMP population also revealed a higher proportion of isolates carrying more than one plasmid. Assessment of genetic diversity of the native rhizobia populations by a DNA fingerprinting PCR method, here designated as DAPD (Direct Amplified Polymorphic DNA), showed a higher degree of variation in Elvas-ENMP and Beja populations. The symbiotic effectiveness (SE) of 39 isolates was determined and ranged 13-34%. Statistical analysis showed that SE was negatively correlated with plasmid number of the isolate. Conclusions: The largest indigenous rhizobia population was found in Elvas-ENMP. DAPD pattern and plasmid profile analysis both suggested a higher genetic diversity among the populations of Elvas-ENMP and Beja. No relationship was found between SE of the isolates and their origin site. Significance and Impact of Study: The large native population, rather than the symbiotic performance of individual rhizobia, could contribute to the higher chickpea yields obtained in Elvas-ENMP

Seasonal patterns of polycyclic aromatic hydrocarbons in digestive gland and arm of octopus (Octopus vulgaris) from the Northwest Atlantic

Among organic pollutants existing in coastal areas, polycyclic aromatic hydrocarbons (PAHs) are of great concern due to their ubiquity and carcinogenic potential. The aim of this study was to evaluate the seasonal patterns of PAHs in the digestive gland and arm of the common octopus (Octopus vulgaris) from the Northwest Atlantic Portuguese coast. In the different seasons, 18 PAHs were determined and the detoxification capacity of the species was evaluated. Ethoxyresorufin O-deethylase (EROD) and ethoxycoumarin O-deethylase (ECOD) activities were measured to assess phase I biotransformation capacity. Individual PAH ratios were used for major source (pyrolytic/petrogenic) analysis. Risks for human consumption were determined by the total toxicity equivalence approach. Generally, low levels of PAHs were detected in the digestive gland and in the arm of octopus, with a predominance of low molecular over high molecular weight compounds. PAHs exhibited seasonality in the concentrations detected and in their main emission sources. In the digestive gland, the highest total PAH levels were observed in autumn possibly related to fat availability in the ecosystem and food intake. The lack of PAH elimination observed in the digestive gland after captivity could be possibly associated to a low biotransformation capacity, consistent with the negligible/undetected levels of EROD and ECOD activity in the different seasons. The emission sources of PAHs found in the digestive gland varied from a petrogenic profile observed in winter to a pyrolytic pattern in spring. In the arm, the highest PAH contents were observed in June; nevertheless, levels were always below the regulatory limits established for food consumption. The carcinogenic potential calculated for all the sampling periods in the arm were markedly lower than the ones found in various aquatic species from different marine environments. The results presented in this study give relevant baseline data for environmental monitoring of organic pollution in coastal areas

Detecção de Rickettsias do grupo febre maculosa em cães e equinos em São José dos Pinhais, PR

Orientador: Prof. Marcelo Beltrão MolentoDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Agrárias, Programa de Pós-Graduação em Ciencias Veterinárias. Defesa: Curitiba, 2007Inclui bibliografiaA Febre Maculosa Brasileira (FMB) é uma zoonose de caráter endêmico, com casos humanos registrados em grande parte do território nacional. Entretanto o Paraná teve o primeiro registro da doença em abril de 2005, no município de São José dos Pinhais, região metropolitana de Curitiba. Com o objetivo de iniciar estudos da epidemiologia da FMB na região supracitada, foram colhidas amostras de sangue de eqüinos de carroceiros da região, através de adesão voluntária dos proprietários, e também de eqüinos e cães provenientes de 4 áreas consideradas foco (um foco humano e três focos animais). As amostras de um total de 83 eqüinos e 16 caninos foram processadas pela técnica de Reação de Imunofluorescência Indireta (RIFI), que é considerada padrão ouro para diagnóstico sorológico da FMB. Para as amostras dos eqüinos de carroceiros foram utilizados antígenos de Rickettsia rickettsii, e para as amostras dos animais dos focos, R. rickettsii e R. parkeri. Os resultados foram de soropositividade em 9,33% dos eqüinos de carroceiros. Dos eqüinos dos focos, houve positividade em 25% de soros para ambos os antígenos testados, e dos cães, 12,5% de soros positivos, todos apenas para R. rickettsii. Estes resultados representam necessidade de avaliação constante da FMB na região e devem ser complementados com pesquisa de positividade nos vetores do gênero Amblyomma sp. por meio de biologia molecularBrazilian spotted fever (BSF) is an endemic zoonosis and human cases have been registered in almost all country. However, the first registered case in Paraná State was in April of 2005, in São José dos Pinhais, suburb area of Curitiba. In order to begin the studies about BSF epidemiology in this area, it was obtain serum samples of cart horses, by owners volunteer adhesion, and of horses and dogs in 4 focus (a human focus and three animal focus). Eighty three horses and sexteen dogs samples were prossessing by Indirect Immunofluorescence Assay (IFA), the gold standart test for serologic diagnostic of BSF. Cart horse samples were done using an antigen against Rickettsia rickettsii, and for the focus animals it was used R. rickettsii and R. parkeri antigens. It was determined soropositivity in 9,33% of cart horses. In focus horses 25% was positive to both antigens, and 12,5% of dogs were positive to R. rickettsii. The results demonstrate the necessity for constant evaluation of BSF in this area, and should be complemented whit positivity vectors of Amblyomma sp. gennus investigation by molecular biolog