Molecular and serological characterization of Leptospira kirschneri serogroup Pomona isolated from a human case in a Brazilian rural area

Submitted by Sandra Infurna ([email protected]) on 2017-11-30T11:27:51Z No. of bitstreams: 1 juliana_vitalbrasil_etal_IOC_2017.pdf: 1011010 bytes, checksum: 4ea03b48249a25f91b3c7a098c03779d (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2017-11-30T11:35:59Z (GMT) No. of bitstreams: 1 juliana_vitalbrasil_etal_IOC_2017.pdf: 1011010 bytes, checksum: 4ea03b48249a25f91b3c7a098c03779d (MD5)Made available in DSpace on 2017-11-30T11:35:59Z (GMT). No. of bitstreams: 1 juliana_vitalbrasil_etal_IOC_2017.pdf: 1011010 bytes, checksum: 4ea03b48249a25f91b3c7a098c03779d (MD5) Previous issue date: 2017Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Zoonoses Bacterianas. Centro de Referência Nacional para Leptospirose. World Health Organization/Pan American Health Organization. Centro Colaborador para Leptospirose - Coleção de Leptospira. Rio de Janeiro, RJ. Brasil..Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Zoonoses Bacterianas. Centro de Referência Nacional para Leptospirose. World Health Organization/Pan American Health Organization. Centro Colaborador para Leptospirose - Coleção de Leptospira. Rio de Janeiro, RJ. Brasil..Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Zoonoses Bacterianas. Centro de Referência Nacional para Leptospirose. World Health Organization/Pan American Health Organization. Centro Colaborador para Leptospirose - Coleção de Leptospira. Rio de Janeiro, RJ. Brasil..Fundação Estadual de Produção e Pesquisa em Saúde. Instituto de Pesquisas Biológicas. Laboratório Central. Porto Alegre, RS, Brasil..Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Zoonoses Bacterianas. Centro de Referência Nacional para Leptospirose. World Health Organization/Pan American Health Organization. Centro Colaborador para Leptospirose - Coleção de Leptospira. Rio de Janeiro, RJ. Brasil..Leptospirosis is an important health concern in Brazil. Currently, information on the epidemiology of the disease in the rural areas of the country is lacking

Aplicação da proteína ErpY-like em IgG-ELISA indireto para o diagnóstico da leptospirose em humanos

A leptospirose é uma antropozoonose difundida mundialmente, muito comum em regiões tropicais e em países em desenvolvimento, acometendo animais domésticos, silvestres e também humanos. Atualmente, o padrão ouro para diagnóstico da leptospirose é o teste de aglutinação microscópica (MAT), método este reconhecido pela Organização Mundial da Saúde (OMS). Contudo, esse método é laborioso e demorado e, além disso, pode levar a resultados errôneos em consequência de diferentes interpretações. Recentemente nosso grupo de pesquisa demonstrou que ErpY-like protege hamsters contra infecção letal (OLIVEIRA et al., 2018) e é capaz de diagnosticar a infecção em suínos, quando empregada em um teste sorológico do tipo ELISA (PADILHA et al., 2019). Estes resultados encorajam o prosseguimento das pesquisas, objetivando o desenvolvimento de novos insumos para o controle da enfermidade, que sejam fáceis de utilizar pelos serviços de saúde, mais sensíveis e específicos, e que empreguem tecnologia nacional, diminuindo custos. Diante disso, o presente estudo buscou avaliar o potencial da proteína recombinante ErpY-like de L. interrogans, no desenvolvimento de um teste do tipo ELISA (Enzyme-Linked Immunosorbent Assay), para detecção de anticorpos IgG no soro de humanos naturalmente infectados

Multiplex PCR-based detection of Leptospira in environmental water samples obtained from a slum settlement

The aim of this study was to apply a molecular protocol to detect leptospiral DNA in environmental water samples. The study was carried out in a peri-urban settlement in Petrópolis, state of Rio de Janeiro. A multiplex PCR method employing the primers LipL32 and 16SrRNA was used. Three out of 100 analysed samples were positive in the multiplex PCR, two were considered to have saprophytic leptospires and one had pathogenic leptospires. The results obtained supported the idea that multiplex PCR can be used to detect Leptospira spp in water samples. This method was also able to differentiate between saprophytic and pathogenic leptospires and was able to do so much more easily than conventional methodologies

The involvement of tetA and tetE tetracycline resistance genes in plasmid and chromosomal resistance of Aeromonas in Brazilian strains

This study analyzed the involvement of tetA and tetE genes in the tetracycline resistance of 16 strains of genus Aeromonas, isolated from clinical and food sources. Polymerase chain reactions revealed that 37.5% of the samples were positive for tetA, and also 37.5% were tetE positive. One isolate was positive for both genes. Only the isolate A. caviae 5.2 had its resistance associated to the presence of a plasmid, pSS2. The molecular characterization of pSS2 involved the construction of its restriction map and the determination of its size. The digestion of pSS2 with HindIII originated two fragments (A and B) that were cloned separately into the pUC18 vector. The tetA gene was shown to be located on the HindIII-A fragment by PCR. After transforming a tetracycline-sensitive strain with pSS2, the transformants expressed the resistance phenotype and harbored a plasmid whose size was identical to that of pSS2. The results confirmed the association between pSS2 and the tetracycline resistance phenotype, and suggest a feasible dissemination of tetA and tetE among strains of Aeromonas. This study suggests the spreading tetA and tetE genes in Aeromonas in Brazil and describes a resistance plasmid that probably contributes to the dissemination of the resistance

Seroprevalence of and risk factors for leptospirosis in the City of Manaus, State of Amazonas, Brazil

Abstract INTRODUCTION Leptospirosis is caused by a bacterium of the genus Leptospira. This study aimed at investigating the seroprevalence of and risk factors for leptospirosis in humans in Manaus, State of Amazonas. METHODS Interviews were performed, and 1,000 blood serum samples were examined using a microscopic agglutination test. RESULTS Forty-three cases were positive; there were 10 serotypes, with coagglutination in 8 cases. The most frequently occurring serotypes were Icterohaemorrhagiae (20.7%), Cynopteri (20.7%), Australis (18.8%), and Copenhageni (16.9%), and the Midwest (54.7%) and South (23.8%) had the most cases; these areas lack basic sanitation. CONCLUSIONS Disease occurrence might be reduced through improved basic infrastructural conditions

Characterization of Leptospira sp reference strains using the pulsed field gel electrophoresis technique

Submitted by Sandra Infurna ([email protected]) on 2018-03-13T16:47:33Z No. of bitstreams: 1 rachel_riberio_etal_IOC_2010.pdf: 677571 bytes, checksum: 6a76dba0531d5351c61bda6b4254c354 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2018-03-13T16:55:10Z (GMT) No. of bitstreams: 1 rachel_riberio_etal_IOC_2010.pdf: 677571 bytes, checksum: 6a76dba0531d5351c61bda6b4254c354 (MD5)Made available in DSpace on 2018-03-13T16:55:10Z (GMT). No. of bitstreams: 1 rachel_riberio_etal_IOC_2010.pdf: 677571 bytes, checksum: 6a76dba0531d5351c61bda6b4254c354 (MD5) Previous issue date: 2010Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Referência Nacional para Leptospirose. Centro Colaborador da Organização Mundial da Saúde. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Referência Nacional para Leptospirose. Centro Colaborador da Organização Mundial da Saúde. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Referência Nacional para Leptospirose. Centro Colaborador da Organização Mundial da Saúde. Rio de Janeiro, RJ. Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Referência Nacional para Leptospirose. Centro Colaborador da Organização Mundial da Saúde. Rio de Janeiro, RJ. Brasil.Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Professor Paulo de Góes. Rio de Janeiro, RJ, BrasilUniversidade Federal do Rio de Janeiro. Instituto de Microbiologia Professor Paulo de Góes. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Referência Nacional para Leptospirose. Centro Colaborador da Organização Mundial da Saúde. Rio de Janeiro, RJ. Brasil.A leptospirose é uma zoonose endêmica, mundialmente distribuída, causada por bactérias do gênero Leptospira. Este gênero compreende espécies patogênicas e saprofíticas, com mais de 200 sorovares distintos, dificultando sua caracterização. A técnica de pulsed field gel electrophoresis tem sido empregada como uma ferramenta para auxiliar nesta caracterização. Os objetivos deste trabalho foram padronizar a técnica de PFGE, determinar os perfis moleculares das cepas de referência utilizadas pelo Laboratório de Referência Nacional para Leptospirose/Centro Colaborador da Organização Mundial de Saúde para Leptospirose e criar um banco de dados com estes perfis. Métodos: Foram analisadas, por PFGE, dezenove cepas utilizando a enzima de restrição NotI. Resultados: Cada cepa apresentou um perfil único que pode ser considerado como uma identidade genômica específica, com exceção dos sorovares Icterohaemorrhagiae e Copenhageni, cujos perfis foram indistinguíveis. Conclusões: Dessa forma, foi possível a criação de um banco de perfis moleculares que está sendo utilizado no Laboratório para a comparação e identificação de cepas isoladas de quadros clínicos.Leptospirosis is an endemic zoonosis of worldwide distribution, caused by bacteria of the genus Leptospira. This genus includes pathogenic and saprophytic species, with more than 200 different serovars, thus making it difficult to characterize. The technique of pulsed field gel electrophoresis has been used as a tool to aid in this characterization. The aims of this study were to standardize the PFGE technique, determine the molecular profiles of reference strains used at the National Reference Laboratory for Leptospirosis/World Health Organization Collaborating Center for Leptospirosis and create a database with these profiles. Methods: Nineteen strains were analyzed by means of PFGE, using the restriction enzyme NotI. Results: Each strain presented a unique profile that could be considered to be a specific genomic identity, with the exception of the serovars Icterohaemorrhagiae and Copenhageni, whose profiles were indistinguishable. Conclusions: It was possible to create a database of molecular profiles, which are being used in the Laboratory for comparing and identifying strains isolated from clinical cases

Characterization of Leptospira isolates from humans and the environment in Uruguay

ABSTRACT Laboratory diagnosis of human leptospirosis usually relies on indirect methods exploring specific immune response. Isolation and identification of the involved strains are cumbersome, but can provide biological resources for pathogenic studies and relevant information for guiding prevention and control measures. The aim of the research we are hereby reporting was the characterization of Leptospira isolates obtained from humans and the environment in Uruguay. Blood cultures were performed from early samples of 302 Uruguayan patients, mainly rural workers, and from 36 water samples taken from their living or working environments. Eight human isolates and seven environmental isolates were obtained and analyzed by end point Polymerase Chain Reaction (PCR), Multilocus Variable Number of Tandem Repeat Analysis (MLVA) and other molecular methods. Human isolates corresponded to several serogroups and serovars of Leptospira interrogans and Leptospira kirschneri species, probably reflecting the infection with similar involved Leptospira species and serovars of an extended animal reservoir in rural settings of the country, mostly dedicated to meat and dairy production. Culture-positive patients were older than usually affected workers, and presented signs and symptoms of severe illness. A high organic and circulating bacterial burden may explain an easier positive result from these workers’ samples. Environmental isolates were mainly identified as Leptospira biflexa strains, with a single L. meyeri isolate of uncertain significance