Polymerase chain reaction tests for the identification of Ross River, Kunjin and Murray Valley encephalitis virus infections in horses

Objective To develop and validate specific, sensitive and rapid diagnostic tests using RT-PCR for the detection of Ross River virus (RRV), Kunjin virus (KV) and Murray Valley encephalitis virus (MVEV) infections in horses. Methods Primer sets based on nucleotide sequence encoding the envelope glycoprotein E2 of RRV and on the nonstructural protein 5 (NS5) of KV and MVEV were designed and used in single round PCRs to test for the respective viruses in infected cell cultures and, in the case of RRV, in samples of horse blood and synovial fluid. Results The primer pairs designed for each of the three viruses amplified a product of expected size from prototype viruses that were grown in cell culture. The identity of each of the products was confirmed by nucleotide sequencing indicating that in the context used the RT-PCRs were specific. RRV was detected in serums from 8 horses for which there were clinical signs consistent with RRV infection such that an acute-phase serum sample was taken and submitted for RRV serology testing. The RRV RT-PCR was analytically sensitive in that it was estimated to detect as little as 50 TCID50 of RRV per mL of serum and was specific in that the primer pairs did not amplify other products from the 8 serum samples. The RRV primers also detected virus in three independent mosquito pools known to contain RRV by virus isolation in cell culture. Samples from horses suspected to be infected with KV and MVEV were not available. Conclusion Despite much anecdotal and serological evidence for infection of horses with RRV actual infection and associated clinical disease are infrequently confirmed. The availability of a specific and analytically sensitive RT-PCR for the detection of RRV provides additional opportunities to confirm the presence of this virus in clinical samples. The RTPCR primers for the diagnosis of KV and MVEV infections were shown to be specific for cell culture grown viruses but the further validation of these tests requires the availability of appropriate clinical samples from infected horses.

A Educação Ambiental nos encontros do Congo com os cotidianos escolares de uma Escola Municipal da Barra do Jucu, Vila Velha, ES

A pesquisa “A Educação Ambiental nos encontros do Congo com os cotidianos escolares de uma Escola Municipal da Barra do Jucu, Vila Velha, ES” se apoia na produção narrativa em Educação Ambiental com os cotidianos, com a intenção de problematizar os saberes e fazeres socioambientais produzidos nos encontros entre a produção cultural do congo com as práticas escolares cotidianas. Com as conversas e narrativas dos sujeitos da pesquisa, alunos(as), professores(as), congueiros(as), pais de alunos e outros representantes da comunidade escolar, problematizamos e mapeamos esses saberesfazeres socioambientais, bem como compreendemos suas diferentes traduções. Os cotidianos escolares são considerados espaços de burla às hierarquizações culturais, ambientais, sociais e curriculares, por serem espaços privilegiados de práticas curriculares que vão além das propostas instituídas, e enfatizamos como o entorno da escola com suas especificidades locais se constituem potências para a produção de saberes e a criação de currículos cotidianamente. As oficinas de congo que acontecem na Escola Municipal da Barra do Jucu foram acompanhadas e mostraram-se potentes para o desenvolvimento da Educação Ambiental numa perspectiva pós-colonial, que acredita na possibilidade de um pensamento que vá além da relação dicotômica cultura/natureza e da não homogeneização cultural. As produções culturais desenvolvidas nas oficinas possibilitaram “outras” alternativas para a produção de subjetividades e para trocas de saberes socioambientais e culturais, baseados na solidariedade que potencializa práticas sociais sustentáveis.

Entre rios, praias e planetas : travessias do congo da Barra do Jucu

O Espírito Santo abriga grande diversidade de expressões populares tradicionais que se dividem em grupos, saberes e celebrações. Uma das práticas mais evidentes são as bandas de Congo que abrangem grande parte do Estado e ganham cada vez mais notoriedade. A Barra do Jucu é uma das comunidades mais antigas do Espírito Santo, uma vila de pescadores que séculos atrás compunha parte da grande fazenda Araçatiba fundada pelos Jesuítas e administrada no século XIX pelo coronel Sebastião Vieira Machado. A diversidade cultural dessa região habitada por africanos, indígenas e europeus, propiciou o surgimento de práticas populares como a Marujada, a Folia de Reis e as Bandas de Congo, que persistem até hoje nos municípios de Cariacica, Vila Velha, Guarapari e Viana, no passado, pertencentes à grande fazenda Araçatiba. A formação do Congo na Barra do Jucu é resultado de rodas informais realizadas por conguistas de comunidades ribeirinhas vizinhas, promovidas principalmente pelo senhor Ignácio Vieira Machado, descendente do coronel Sebastião e lideradas por Alcides Gomes da Silva, descendente de negros africanos e açorianos habitantes da fazenda Araçatiba. Durante muito tempo essa prática foi marginalizada, mas atualmente o Congo é considerado um ícone da cultura capixaba. Essa ressignificação está ligada a um processo de valorização e projeção midiática que se iniciou nos anos 1980 e se intensificou nas décadas seguintes. Boa parte dos movimentos e eventos que estimularam esse processo ocorreu na comunidade da Barra do Jucu. Por outro lado, políticos, empresários, indústria do entretenimento e outros setores sociais, aproveitam cada vez mais dessa ascensão do Congo utilizando os grupos tradicionais para fins lucrativos e promocionais. De prática marginal até se tornar ícone cultural, o Congo da Barra do Jucu passou por fazendas, rios, praias e planetas. E sobre essas travessias não só geográficas, mas sociais, culturais e políticas é que este trabalho pretende refletir, além das práticas tradicionais dos grupos e seus usos, buscando trazer à tona as relações entre conguistas, instituições públicas e privadas, mídia e público.

ANDRÉ DO COUTO GODINHO: HOMEM PRETO, FORMADO EM COIMBRA, MISSIONÁRIO NO CONGO EM FINS DO SÉCULO XVIII

O fio condutor deste texto é a trajetória de um homem de cor livre nascido na vila de Mariana, no ano de 1720, tendo falecido no Reino do Congo, em data desconhecida, provavelmente no último decênio do século XVIII. Entre as Minas Gerais e o Reino do Congo, vários anos vividos em Coimbra e Lisboa demarcaram a trajetória de André do Couto Godinho. A narrativa, aparentemente excepcional, não se restringe entretanto a episódios da vida pessoal de Godinho, ao contrário, trata de coletivos mais amplos, constituindo-se, quiçá, em uma "janela privilegiada" para o mundo dos homens de cor livres em diferentes localidades do Império português, na segunda metade dos Setecentos. Assim, ao seguir os passos de André Godinho, focalizo contextos históricos que dão sentido e podem, ao mesmo tempo, ser reinterpretados à luz das possibilidades, escolhas e limitações que constituíram sua história pessoal.

Development of a simple analytical method for the simultaneousdetermination of paracetamol, paracetamol-glucuronide andp-aminophenol in river water

Paracetamol is among the most worldwide consumed pharmaceuticals. Although its occurrence in the environment is well documented, data about the presence of its metabolites and transformation products is very scarce. The present work describes the development of an analytical method for the simultaneous determination of paracetamol, its principal metabolite (paracetamol-glucuronide) and its main transformation product (p-aminophenol) based on solid phase extraction (SPE) and high performance liquid chromatography coupled to diode array detection (HPLC-DAD). The method was applied to analysis of river waters, showing to be suitable to be used in routine analysis. Different SPE sorbents were compared and the use of two Oasis WAX cartridges in tandem proved to be the most adequate approach for sample clean up and pre-concentration. Under optimized conditions, limits of detection in the range 40–67 ng/L were obtained, as well as mean recoveries between 60 and 110% with relative standard deviations (RSD) below 6%. Finally, the developed SPE-HPLC/DAD method was successfully applied to the analysis of the selected compounds in samples from seven rivers located in the north of Portugal. Nevertheless all the compounds were detected, it was the first time that paracetamol-glucuronide was found in river water at concentrations up to 3.57 μg/L.

A high throughput colorimetric assay of beta-1,3-D-glucans by Congo red dye

Mushroom strains contain complex nutritional biomolecules with a wide spectrum of therapeutic and prophylactic properties. Among these compounds, β-d-glucans play an important role in immuno-modulating and anti-tumor activities. The present work involves a novel colorimetric assay method for β-1,3-d-glucans with a triple helix tertiary structure by using Congo red. The specific interaction that occurs between Congo red and β-1,3-d-glucan was detected by bathochromic shift from 488 to 516 nm (> 20 nm) in UV–Vis spectrophotometer. A micro- and high throughput method based on a 96-well microtiter plate was devised which presents several advantages over the published methods since it requires only 1.51 μg of polysaccharides in samples, greater sensitivity, speed, assay of many samples and very cheap. β-d-Glucans of several mushrooms (i.e., Coriolus versicolor, Ganoderma lucidum, Pleurotus ostreatus, Ganoderma carnosum, Hericium erinaceus, Lentinula edodes, Inonotus obliquus, Auricularia auricular, Polyporus umbellatus, Cordyseps sinensis, Agaricus blazei, Poria cocos) were isolated by using a sequence of several extractions with cold and boiling water, acidic and alkaline conditions and quantified by this microtiter plate method. FTIR spectroscopy was used to study the structural features of β-1,3-d-glucans in these mushroom samples as well as the specific interaction of these polysaccharides with Congo red. The effect of NaOH on triple helix conformation of β-1,3-d-glucans was investigated in several mushroom species.

Optimization of River Water Quality Surveys by Multivariate Analysis of Physicochemical, Bacteriological and Ecotoxicological Data

This study aims to optimize the water quality monitoring of a polluted watercourse (Leça River, Portugal) through the principal component analysis (PCA) and cluster analysis (CA). These statistical methodologies were applied to physicochemical, bacteriological and ecotoxicological data (with the marine bacterium Vibrio fischeri and the green alga Chlorella vulgaris) obtained with the analysis of water samples monthly collected at seven monitoring sites and during five campaigns (February, May, June, August, and September 2006). The results of some variables were assigned to water quality classes according to national guidelines. Chemical and bacteriological quality data led to classify Leça River water quality as “bad” or “very bad”. PCA and CA identified monitoring sites with similar pollution pattern, giving to site 1 (located in the upstream stretch of the river) a distinct feature from all other sampling sites downstream. Ecotoxicity results corroborated this classification thus revealing differences in space and time. The present study includes not only physical, chemical and bacteriological but also ecotoxicological parameters, which broadens new perspectives in river water characterization. Moreover, the application of PCA and CA is very useful to optimize water quality monitoring networks, defining the minimum number of sites and their location. Thus, these tools can support appropriate management decisions.

Occurrence of Cryptosporidium oocysts and Giardia cysts in raw water from the Atibaia river, Campinas, Brazil

Cryptosporidium parvum and Giardia duodenalis are waterborne parasites that have caused several outbreaks of gastrointestinal disease associated with drinking water. Due to the lack of studies about the occurrence of these protozoa in water in the Southeast of Brazil, an investigation was conducted to verify the presence of cysts and oocysts in superficial raw water of the Atibaia River. The water samples were submitted to membrane filtration (3.0 mum) and elution was processed by (1) scraping and rinsing of membrane (RM method) and (2) acetone-dissolution (ADM method). Microbiologic and chemical parameters were analyzed. Aliquots of the pellets were examined by immunofluorescence (Merifluor, Meridian Diagnostics, Cincinnati, Ohio). All water samples were positive for Cryptosporidium and Giardia, in spite of the high turbidity. Higher recovery rates occurred in samples treated by the RM method than by the ADM technique. The goal for future work is the assessment of viability of cysts and oocysts to determine the public health significance of this finding.

Detection of Vibrio parahaemolyticus and Vibrio cholerae in oyster, Crassostrea rhizophorae, collected from a natural nursery in the Cocó river estuary, Fortaleza, Ceará, Brazil

Oysters are edible organisms that are often ingested partially cooked or even raw, presenting therefore a very high risk to the consumers' health, especially in tropical regions. The presence of Vibrio cholerae and Vibrio parahaemolyticus in oysters sampled at an estuary in the Brazilian northeastern region was studied, with 300 oysters tested through an 8-months period. The salinity of the water at the sampling point varied between 3% and 27‰. V. cholerae was the most frequently detected species (33.3% of the samples), and of the 22 V. cholerae isolates, 20 were identified as non-O1/non-O139, with two of the colonies presenting a rough surface and most of remaining ones belonging to the Heiberg II fermentation group. V. parahaemolyticus was isolated from just one of the samples. Other bacteria such as Providencia spp., Klebsiella spp. and Morganella morganii were also isolated.

SEROLOGICAL DETECTION OF HEPATITIS A VIRUS IN FREE-RANGING NEOTROPICAL PRIMATES (Sapajus spp., Alouatta caraya) FROM THE PARANÁ RIVER BASIN, BRAZIL

Nonhuman primates are considered as the natural hosts of Hepatitis A virus (HAV), as well as other pathogens, and can serve as natural sentinels to investigate epizootics and endemic diseases that are of public health importance. During this study, blood samples were collected from 112 Neotropical primates (NTPs) (Sapajus nigritus and S. cay, n = 75; Alouatta caraya, n = 37) trap-captured at the Paraná River basin, Brazil, located between the States of Paraná and Mato Grosso do Sul. Anti-HAV IgG antibodies were detected in 4.5% (5/112) of NTPs, specifically in 6.7% (5/75) of Sapajus spp. and 0% (0/37) of A. caraya. In addition, all samples were negative for the presence of IgM anti-HAV antibodies. These results suggest that free-ranging NTPs were exposed to HAV within the geographical regions evaluated.

Jean Renoir. The River: Uma história ou um lugar? Passagem pela Índia

Dissertação apresentada para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Ciências da Comunicação

Fluvial system restoration – case study: river Tua

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente, Perfil de Engenharia de Sistemas Ambientais