Culturable bacterial community of the estuarine surface microlayer

Mestrado em MicrobiologiaA camada superficial aquática (1-1000 μm) é um ecossistema único, definido como a interface entre a hidrosfera e a atmosfera. É uma camada exposta a altas intensidades de radiação solar Ultra-Violeta, sendo enriquecida com compostos orgânicos e poluentes antropogénicos. Além disso, está sujeita a condições instáveis de temperatura e salinidade. Assim sendo, é razoável colocar-se a hipótese de que esta camada é habitada por comunidades bacterianas distintas e especializadas. Apenas alguns estudos sobre este tema foram publicados e os resultados foram frequentemente divergentes. Apesar do já reconhecido enviesamento introduzido pelas metodologias dependentes do cultivo, tais técnicas permanecem essenciais para a compreensão da fisiologia e ecologia da comunidade bacteriana. Os estuários são ambientes confinados e frequentemente muito poluídos, o que provavelmente favorece a formação de camadas superficiais claramente distintas das águas subjacentes. Portanto, o objectivo deste trabalho foi comparar as comunidades bacterianas cultiváveis da camada superficial aquática e da coluna de água. Foram escolhidos três locais ao longo do estuário Ria de Aveiro atendendo a diferentes parâmetros ambientais e exposição a poluentes. A amostragem foi realizada utilizando o método 'Glass- Plate'. As amostras foram obtidas em maré baixa, durante o dia e noite, em cinco campanhas, tendo em vista a quantificação das unidades formadoras de colónias e subsequente isolamento para caracterização filogenética. Para estes fins, usámos dois meios de cultura: GSP (Pseudomonas Aeromonas Selective Agar Base) e EA (Estuarine Agar). A quantificação das UFC indica que o número de bactérias provenientes da camada superficial (bacterioneuston) é cerca de três vezes mais abundante do que o proveniente da coluna de água (bacterioplâncton). Verifica-se uma diminuição da abundância de bacterioneuston de dia para noite, ao contrário do bacterioplâncton, que tende a aumentar durante o mesmo período. Dos isolados obtidos, o rDNA 16S foi e digerido com a enzima HaeIII. A partir de 402 isolados, foram identificados 72 perfis diferentes. Desses, 21 perfis foram exclusivos da camada superficial e 28 foram exclusivos da coluna de água. Representantes dos diferentes perfis foram analisados por sequenciação e bactérias pertencentes a 5 Filos: Proteobacteria, Bacteroidetes, Actinobacteria, Firmicutes e Deinococci-Thermus; e 9 Classes: Gammaproteobacteria, Alphaproteobacteria, Betaproteobacteria, Epsilonproteobacteria, Actinobacteria, Flavobacteria, Sphingobacteria, Deinococci e Bacilli foram identificadas. Os isolados afiliaram com sequências provenientes de ambientes aquáticos bem como de áreas altamente contaminadas. Os resultados apontam para uma comunidade cultivável distinta/particular na microcamada superficial estuarina. ABSTRACT: The sea surface microlayer (SML) is an unique ecosystem, defined as the interfacial film (uppermost 1–1000 μm) between the atmosphere and the ocean. Thereby, it is exposed to high intensities of solar radiation, and is enriched with organic compounds and pollutants from anthropogenic inputs. Also it is subjected to unstable temperature and salinity conditions. Thus, it is proper to hypothesize that the SML is inhabited by distinct and specialized microbial communities. Only a few studies on this topic were published and results wee frequently divergent. Despite the previously recognized biases introduced by culture-dependent methodologies, such techniques remain essential to understand bacterial population’s physiology and ecology. Estuaries are confined and frequently highly polluted environments, which probably favor the formation of distinct surface layers clearly distinct from underlying waters. Therefore, our goal was to compare the culturable bacterial communities occurring in SML and underlying waters (UW). Our work concerned three sampling sites in the estuary Ria de Aveiro, corresponding to different environmental parameters and exposure to pollutants. Sampling was conducted using the so-called ‘Glass-Plate’ method. The UW samples were collected directly into a sterilized glass bottle from a depth of approximately 0.4 m. Samples were obtained at low-tide, during day and night, in five campaigns, regarding the CFU (Colony Forming Units) quantification and subsequent recovery of bacterial isolates. For these purposes we used two culture media: GSP (Pseudomonas Aeromonas Selective Agar Base) and EA (Estuarine Agar). CFU quantification indicates that bacterioneuston is about three times more abundant than bacterioplankton. Generally bacterioneuston abundance decreases from day to night while bacterioplankton usually increases during the same period. From all the obtained isolates the 16S rDNA was amplified using universal primers and digested with the enzyme HaeIII. The profiles were analyzed using the software GelCompar and representatives of each pattern were selected for sequencing. From 402 isolates, 72 different profiles were identified. From those 21 profiles were exclusive from SML samples and 28 were exclusive from UW samples. Sequencing results allowed identifying bacteria belonging to 5 different Phyla: Proteobacteria, Bacteroidetes, Actinobacteria, Firmicutes and Deinococci-Thermus; and 9 Classes: Gammaproteobacteria, Alphaproteobacteria, Betaproteobacteria, Epsilonproteobacteria, Actinobacteria, Flavobacteria, Sphingobacteria, Deinococci e Bacilli. Isolates affiliated with sequences from aquatic environments as well as highly contaminated areas. The results point to a distinct/particular culturable community within the SML of this estuarine environment

Development of cross-resistance by Aspergillus fumigatus to clinical azoles following exposure to prochloraz, an agricultural azole

Background: The purpose of this study was to unveil whether azole antifungals used in agriculture, similar to the clinical azoles used in humans, can evoke resistance among relevant human pathogens like Aspergillus fumigatus, an ubiquitous agent in nature. Additionally, cross-resistance with clinical azoles was investigated. Antifungal susceptibility testing of environmental and clinical isolates of A. fumigatus was performed according to the CLSI M38-A2 protocol. In vitro induction assays were conducted involving daily incubation of susceptible A. fumigatus isolates, at 35°C and 180 rpm, in fresh GYEP broth medium supplemented with Prochloraz (PCZ), a potent agricultural antifungal, for a period of 30 days. Minimal inhibitory concentrations (MIC) of PCZ and clinical azoles were monitored every ten days. In order to assess the stability of the developed MIC, the strains were afterwards sub-cultured for an additional 30 days in the absence of antifungal. Along the in vitro induction process, microscopic and macroscopic cultural observations were registered. Results: MIC of PCZ increased 256 times after the initial exposure; cross-resistance to all tested clinical azoles was observed. The new MIC value of agricultural and of clinical azoles maintained stable in the absence of the selective PCZ pressure. PCZ exposure was also associated to morphological colony changes: macroscopically the colonies became mostly white, losing the typical pigmentation; microscopic examination revealed the absence of conidiation. Conclusions: PCZ exposure induced Aspergillus fumigatus morphological changes and an evident increase of MIC value to PCZ as well as the development of cross-resistance with posaconazole, itraconazole and voriconazole.IFR and IMM are supported by FCT (Fundação Ciência e Tecnologia). IFR is supported by FCT PhD grant (SFRH/BD/91155/2012). I.MM is supported by FCT, Ciência 2008 and co-financed by the European Social Fund. The authors would like to thank Isabel Santos for the excellent technical assistance. Part of the results was presented at the 52nd Interscience Conference on Antimicrobial Agents and Chemotherapy, ICAAC, held San Francisco, USA, September 2012.info:eu-repo/semantics/publishedVersio

Environmental azole fungicide, prochloraz, can induce cross-resistance to medical triazoles in Candida glabrata

Acquisition of azole resistance by clinically relevant yeasts in nature may result in a significant, yet undetermined, impact in human health. The main goal of this study was to assess the development of cross-resistance between agricultural and clinical azoles by Candida spp. An in vitro induction assay was performed, for a period of 90 days, with prochloraz (PCZ) - an agricultural antifungal. Afterward, the induced molecular resistance mechanisms were unveiled. MIC value of PCZ increased significantly in all Candida spp. isolates. However, only C. glabrata developed cross-resistance to fluconazole and posaconazole. The increased MIC values were stable. Candida glabrata azole resistance acquisition triggered by PCZ exposure involved the upregulation of the ATP binding cassette multidrug transporter genes and the transcription factor, PDR1. Single mutation previously implicated in azole resistance was found in PDR1 while ERG11 showed several synonymous single nucleotide polymorphisms. These results might explain why C. glabrata is so commonly less susceptible to clinical azoles, suggesting that its exposure to agricultural azole antifungals may be associated to the emergence of cross-resistance. Such studies forward potential explanations for the worldwide increasing clinical prevalence of C. glabrata and the associated worse prognosis of an infection by this species.info:eu-repo/semantics/publishedVersio

A oxigenoterapia nasal de alto fluxo na pessoa com infeção por SARS-CoV-2 – uma scoping review

Introduction: the SARS-CoV-2 infection has come to change medical health care by pressing for the testing of innovative oxygen administration technics. The high flow nasal cannula therapy (HFNC) stood out in this pandemic, as rather conflicting approaches were taken when caring for COVID-19 patients. Objectives: the purpose of this scoping review is to map available data on the benefits of HFNC in patients with SARS-CoV-2 infection. Material and methods: this scoping review will have Joanna Briggs Institute methodology as guidance. Some criteria of eligibility were taken into consideration to define the population, concept and context, taking into account adults with SARS-CoV-2 (population), high flow nasal cannula (concept) and hospital setting (context). Literature published and not publish in Portuguese, English and Spanish was reviewed, specifically from the year of 2020, and from the following databases: MEDLINE complete, CINAHL complete, PubMed, Cochrane, Nursing & Allied Health Collection and OpenGrey and RCAAP. Two independent reviewers screened the titles and summaries, and latter analysed the complete text of the selected documents, using the relevant information. Results: during the course of this search, 119 articles were found of which 12 were included in this scoping review. In general, all studies presented evidences of the benefits of high flow nasal cannula therapy when caring for patients with SARS-CoV-2 infections, as the reduction of the rate of mechanical ventilation; time of hospital stay; better tolerance of the technic as well as better comfort when compared to other oxygen administration technics. Risk in aerosolization was considered to be reduced provided that specific security measurements are secured. Conclusions: the HFNC in considered to be a secure technic with specific benefits in the care of the SARS-CoV-2 infected patient. We note that nurses have a rather crucial role in the supervision and safety of the HFNC patient.Introdução: a infeção por SARS-CoV-2 veio modificar os cuidados de saúde, levando a que técnicas inovadoras de administração de oxigénio fossem testadas. A Oxigenoterapia nasal de alto fluxo (ONAF) ganhou destaque nesta pandemia, tornando-se, muitas vezes, controversa na abordagem ao doente com COVID-19. Objetivos: o objetivo desta scoping review é mapear a evidência disponível sobre os benefícios do uso da ONAF no doente com infeção por SARS-CoV-2.  Materiais e métodos: a presente scoping review será guiada segundo a metodologia do Joanna Briggs Institute. Foram definidos critérios de elegibilidade com base na população, conceito e contexto, considerando adultos com infeção por SARS-CoV-2 (população), oxigenoterapia nasal de alto fluxo (conceito), contexto hospitalar (contexto). Considerou-se bibliografia publicada e não publicada em português, inglês e espanhol, sendo limitada para publicações do ano 2020, nas bases de dados MEDLINE complete, CINAHL complete, PubMed, Cochrane, Nursing & Allied Health Collection e ainda OpenGrey e RCAAP. Dois revisores independentes triaram os títulos e resumos e posteriormente analisaram o texto completo dos documentos selecionados, extraindo a informação útil. Resultados: pela pesquisa em bases de dados encontraram-se 119 artigos, tendo sido apenas incluídos 12 na scoping review. Na generalidade, todos os estudos apresentam benefícios acerca da utilização da ONAF na pessoa com infeção por SARS-CoV-2, nomeadamente na redução da taxa de ventilação mecânica, na redução do tempo de internamento, uma melhor tolerância da técnica, bem como mais conforto, comparativamente a outras terapias de administração de oxigénio. Considera-se que o risco de contaminação por aerossolização é reduzido, caso sejam garantidas as devidas medidas de segurança. Conclusões: a ONAF é considerada uma técnica segura e com benefícios na abordagem ao doente com infeção por SARS-CoV-2. Ressalvamos que os enfermeiros têm um papel fundamental na vigilância e segurança da pessoa submetida à ONAF

Confiabilidade da informação sobre antecedente de varicela em crianças pré-escolares

OBJECTIVE: To verify how reliable is the information provided by parents about the history of varicella in their children. METHODS: 204 parents of previously healthy children attending two municipal day-care centers of São Paulo city were interviewed between August 2003 and September 2005. A standardized form was filled out with information regarding age, sex, history of varicella and other diseases, drug use and antecedent of immunization, After medical history, physical examination and checking of immunization records, 5 ml of blood were collected for ELISA (in house) varicella test. Exclusion criteria were: age less than 1 year or more than 60 months, previous immunization against chickenpox, presence of co-morbidities or recent use of immunosuppressive drugs. Data were filed in a data bank using the Excel 2003 Microsoft Office Program and stored in a PC computer. The exact Fisher test was employed to calculate sensibility, specificity, positive and negative predictive values of history of varicella informed by children's parents. RESULTS: The age of the children varied from 12 to 54 months (median, 26 months; 49 (24%) children had positive history of varicella, 155 (76%) a negative or doubtful history. The predictive positive and negative values of the information were 90% and 93%, respectively (p = 0.0001). CONCLUSIONS: The degree of reliability of information about history of varicella informed by parents of children attending day care centers was high and useful to establish recommendations on varicella blocking immunization in day-care centers.OBJETIVOS: Verificar o grau de confiabilidade da informação fornecida pelos pais de crianças atendidas em creches sobre o antecedente de varicela. MÉTODOS: Os pais de 204 crianças previamente saudáveis matriculadas em duas creches da cidade de São Paulo foram entrevistados entre Agosto de 2003 e Setembro de 2005 para preenchimento de um questionário padronizado com informações sobre idade, sexo, antecedente de varicela e outras doenças, uso de medicamentos e história vacina. Após anamnese, exame físico e verificação da carteira de vacinação, foram colhidos 5 ml de sangue para realização do teste ELISA (in house) para varicela. Os critérios de exclusão foram: idade < 1 ano ou acima de 60 meses, vacinação prévia contra varicela, presença de comorbidades e uso recente de imunossupressores. Os dados foram registrados num banco de dados do programa Excel 2003 Microsoft Office e armazenados em microcomputador. Utilizou-se o teste exato de Fisher para calcular a sensibilidade, especificidade e valores preditivo positivo e negativo das informações fornecidas pelos pais sobre o antecedente de varicela das crianças. RESULTADOS: A idade das crianças variou entre 12 e 54 meses (mediana, 26 meses); 49 crianças (24%) tinham antecedente positivo para varicela e 155 (76%) negativo ou duvidoso. Os valores preditivo positivo e negativo da informação foram de 90% e 93%, respectivamente (p = 0.0001). CONCLUSÕES: O grau de confiabilidade das informações sobre varicela informadas pelos pais de crianças atendidas em creches foi elevado e útil para estabelecer recomendações para vacinação de bloqueio de surtos de varicela em creches

Splenic autoimplant and hyperbaric oxygen therapy in rats

OBJECTIVE: To study the functional and morphological features of splenic autoimplants in rats subjected, or not, to postoperative hyperbaric oxygen therapy. METHODS: One hundred and five male Wistar rats, weighing 251.6 g ± 29.5 g, were studied ot early (11th day) and late (70th day ) postoperative periods. For each period the animals were distributed in the following groups: A (n=40), splenic manipulation; B (n=65),splenic autoimplants, not treated (nt) (A11nt- n=10, B11nt- n=21, A70nt- n= 10, B70nt-n= 18) and treated with hyperbaric oxygen (t) ( A11t- n= 10, B11t- n=15, A70t - n=10, B70t- n=11). Blood was collected for measurement of lipids and immunoglobulins , platelet and Howell-Jolly body count before and after surgery. The spleen and autoimplants were removed for histologic analyses. RESULTS: There was an increase of total cholesterol (p=0.00068), LDL-cholesterol (p=0.002), VLDL-cholesterol and triglycerides (p=0.0033) in B11nt group. There was an increase of total cholesterol (p= 0.0007) and LDL-cholesterol (p=0.0000) in B70nt group, and no alterations were observed in the other groups. IgM decreased in B groups (p=0.0000) and no changes were observed in the splenic manipulation group. Number of Howell-Jolly bodies were lesser in Bt groups than in Bnt groups( p=0.03). Platelets were increased in B11t and B11nt groups (p=0.0000) and remained unchanged in the other groups. Autoimplants microscopic viability was better in Bt groups than in Bnt. CONCLUSION: The splenic autoimplants of rats treated with hyperbaric oxygen therapy showed a better functionality and viability than those subjected to other procedures.OBJETIVO: Estudar aspectos morfológicos e funcionais dos autoimplantes esplênicos de ratos submetidos ou não à oxigenioterapia hiperbárica (OHB). MÉTODOS: Foram estudados em dois períodos distintos 105 ratos Wistar. No período mediato (n= 56) os animais foram avaliados até o 11º dia de pós-operatório, e no período tardio (n= 49), até o 70º dia. Em cada período os ratos foram distribuídos nos grupos: A- Simulação, B- Autoimplante esplênico, tratados com oxigênio hiperbárico ou não. Nos animais do Grupo A realizou-se apenas manipulação do baço. Nos animais do Grupo B realizou-se esplenectomia total e a seguir quatro fragmentos do baço foram implantados no grande omento. Em todos os animais foram dosados os lípides e imunoglobulinas e contadas as plaquetas e os corpúsculos de Howell-Jolly no pré-operatório e no 11º ou 70º dia de pós-operatório. O baço dos animais do Grupo A e os autoimplantes dos animais do Grupo B foram retirados e enviados para avaliação morfológica. RESULTADOS: No grupo B11nt houve aumento do colesterol total, LDL-colesterol, VLDL-colesterol e triglicérides. No grupo B70nt houve aumento do colesterol total e LDL-colesterol. Nos grupos tratados não houve alterações lipídicas. A IgM caiu nos grupos B e não alterou nos grupos A. Os corpúsculos de Howell - Jolly foram menos freqüentes nos grupos Bt que nos grupos Bnt. As plaquetas aumentaram nos grupos B11t e B11nt e não se alteraram nos demais grupos. A viabilidade microscópica dos grupos Bt foi melhor que a dos grupos Bnt. CONCLUSÃO: Os autoimplantes esplênicos dos animais tratados com OHB apresentaram melhor função e viabilidade do que os autoimplantes dos animais não tratados.22623

Reliability of information on varicella history in preschool children

OBJECTIVE: To verify how reliable is the information provided by parents about the history of varicella in their children. METHODS: 204 parents of previously healthy children attending two municipal day-care centers of São Paulo city were interviewed between August 2003 and September 2005. A standardized form was filled out with information regarding age, sex, history of varicella and other diseases, drug use and antecedent of immunization, After medical history, physical examination and checking of immunization records, 5 ml of blood were collected for ELISA (in house) varicella test. Exclusion criteria were: age less than 1 year or more than 60 months, previous immunization against chickenpox, presence of co-morbidities or recent use of immunosuppressive drugs. Data were filed in a data bank using the Excel 2003 Microsoft Office Program and stored in a PC computer. The exact Fisher test was employed to calculate sensibility, specificity, positive and negative predictive values of history of varicella informed by children's parents. RESULTS: The age of the children varied from 12 to 54 months (median, 26 months; 49 (24%) children had positive history of varicella, 155 (76%) a negative or doubtful history. The predictive positive and negative values of the information were 90% and 93%, respectively (p = 0.0001). CONCLUSIONS: The degree of reliability of information about history of varicella informed by parents of children attending day care centers was high and useful to establish recommendations on varicella blocking immunization in day-care centers.OBJETIVOS: Verificar o grau de confiabilidade da informação fornecida pelos pais de crianças atendidas em creches sobre o antecedente de varicela. MÉTODOS: Os pais de 204 crianças previamente saudáveis matriculadas em duas creches da cidade de São Paulo foram entrevistados entre Agosto de 2003 e Setembro de 2005 para preenchimento de um questionário padronizado com informações sobre idade, sexo, antecedente de varicela e outras doenças, uso de medicamentos e história vacina. Após anamnese, exame físico e verificação da carteira de vacinação, foram colhidos 5 ml de sangue para realização do teste ELISA (in house) para varicela. Os critérios de exclusão foram: idade < 1 ano ou acima de 60 meses, vacinação prévia contra varicela, presença de comorbidades e uso recente de imunossupressores. Os dados foram registrados num banco de dados do programa Excel 2003 Microsoft Office e armazenados em microcomputador. Utilizou-se o teste exato de Fisher para calcular a sensibilidade, especificidade e valores preditivo positivo e negativo das informações fornecidas pelos pais sobre o antecedente de varicela das crianças. RESULTADOS: A idade das crianças variou entre 12 e 54 meses (mediana, 26 meses); 49 crianças (24%) tinham antecedente positivo para varicela e 155 (76%) negativo ou duvidoso. Os valores preditivo positivo e negativo da informação foram de 90% e 93%, respectivamente (p = 0.0001). CONCLUSÕES: O grau de confiabilidade das informações sobre varicela informadas pelos pais de crianças atendidas em creches foi elevado e útil para estabelecer recomendações para vacinação de bloqueio de surtos de varicela em creches

Empire Building and Modernity

Following the organization, in 2009, of the first conference on The British Empire: Ideology, Perspectives, Perception, the Research Group dedicated to Culture Studies at the University of the Lisbon Centre for English Studies organized, in 2010, a second conference under the general title Empire Building and Modernity. This conference constitutes the second part of a three year project undertaken by the group, which will be followed, in 2011, by a third initiative, called Reviewing Imperial Conflicts. The proceedings of the second conference are now presented in this book. Empire Building and Modernity gives a larger scope to the original project, which was developed more strictly around the British Empire, and provides the opportunity to deal with questions related to the formation of modern European empires, namely the Portuguese Colonial Empire. The different chapters in this book reveal a variety of approaches that are very often at the cutting edge of the methodologies adopted in cultural studies, particularly in the field of post-colonial studies. The building of new perceptions on imperial issues interpreted through literature, the visual arts, history and political science, the role of museums, questions of gender and race and the construction of identity through language constitute the guidelines of the contributions presented in this volume. I hope you will enjoy reading it as much as we enjoyed discussing the issues that contributed to its making.Fundação para a Ciência e a Tecnologi