Métodos convencionais e moleculares para tipagem de Salmonella Typhi isoladas no Brasil

Phenotypic and genotypic characteristics of Salmonella Typhi were studied in 30 strains, isolated in different years, from some areas in Brazil. Conventional typing methods were performed by biochemical tests, Vi phage-typing scheme, and antimicrobial susceptibility test. Molecular typing methods were performed by analysis of plasmid DNA and by random amplified polymorphic DNA (RAPD-PCR). For the latter, an optimization step was performed to ensure the reproducibility of the process in genetic characterization of S. Typhi. The predominance of 76.7% of biotype I (xylose +, arabinose -) was noticed in all studied areas. Three phage types were recognized, with prominence for the phage types A (73.3%) and I+IV (23.3%). All the strains were susceptible to the drugs used. However, 36.7% of the strains contained plasmids, with predominance of the 105 Kb plasmid. RAPD was capable of grouping the strains in 8 genotypic patterns using primer 784, in 6, using primer 787 and in 7, using primer 797. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented nonsignificant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and performance, being considered as a promising alternative typing method for S. Typhi.Características fenotípicas e genotípicas de Salmonella Typhi foram estudadas em 30 amostras originárias de certas regiões do Brasil e isoladas em diferentes anos. Os métodos convencionais foram realizados através da tipagem bioquímica, da fagotipagem Vi e do teste de suscetibilidade aos antimicrobianos. Os métodos moleculares foram realizados através das análises do DNA plasmidial e do DNA polimórfico amplificado aleatoriamente (RAPD-PCR). Para este último, uma etapa de otimização foi promovida para garantir a reprodutibilidade do processo na caracterização genotípica das cepas. Foi observada a predominância de 76,7% do biotipo I (xilose +, arabinose -) em todas as regiões consideradas. Três fagotipos foram reconhecidos, com destaque para os fagotipos A (73,3%) e I+IV (23,3%). Todas as amostras demonstraram sensibilidade às drogas testadas. No entanto, 36,7 % das amostras evidenciaram plasmídios, predominando o de 105Kb. RAPD-PCR agrupou as amostras em 8 perfis genotípicos com o iniciador 784, em 6 perfis com o iniciador 787 e em 7, com o iniciador 797. Os métodos convencionais, bem como a análise do DNA plasmidial, não mostraram poder discriminatório significativo; entretanto, a análise por RAPD-PCR mostrou poder discriminatório, reprodutibilidade, fácil interpretação e execução, sendo considerada uma alternativa promissora na tipagem de S. Typhi

Conventional and molecular typing of Salmonella typhi strains from Brazil

Submitted by Sandra Infurna ([email protected]) on 2019-11-14T11:10:55Z No. of bitstreams: 1 BiancaQuintaes_EricaFonseca_etal_IOC_2002.pdf: 131543 bytes, checksum: 6bde448a2a3285a8b4318a5e0b0bbd73 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2019-11-14T11:26:03Z (GMT) No. of bitstreams: 1 BiancaQuintaes_EricaFonseca_etal_IOC_2002.pdf: 131543 bytes, checksum: 6bde448a2a3285a8b4318a5e0b0bbd73 (MD5)Made available in DSpace on 2019-11-14T11:26:03Z (GMT). No. of bitstreams: 1 BiancaQuintaes_EricaFonseca_etal_IOC_2002.pdf: 131543 bytes, checksum: 6bde448a2a3285a8b4318a5e0b0bbd73 (MD5) Previous issue date: 2002Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bacteriologia. Rio de Janeiro, RJ, Brasil / Universidade do Estado do Rio de Janeiro. Departamento de Microbiologia Médica, Curso de Pós-Graduação. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Aggeu Magalhães. Recife, PE, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bacteriologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bacteriologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bacteriologia. Rio de Janeiro, RJ, Brasil.Phenotypic and genotypic characteristics of Salmonella Typhi were studied in 30 strains, isolated in different years, from some areas in Brazil. Conventional typing methods were performed by biochemical tests, Vi phage-typing scheme, and antimicrobial susceptibility test. Molecular typing methods were performed by analysis of plasmid DNA and by random amplified polymorphic DNA (RAPD-PCR). For the latter, an optimization step was performed to ensure the reproducibility of the process in genetic characterization of S. Typhi. The predominance of 76.7% of biotype I (xylose +, arabinose -) was noticed in all studied areas. Three phage types were recognized, with prominence for the phage types A (73.3%) and I+IV (23.3%). All the strains were susceptible to the drugs used. However, 36.7% of the strains contained plasmids, with predominance of the 105 Kb plasmid. RAPD was capable of grouping the strains in 8 genotypic patterns using primer 784, in 6, using primer 787 and in 7, using primer 797. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented nonsignificant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and performance, being considered as a promising alternative typing method for S. Typhi

Padrão de susceptibilidade a antimicrobianos e perfil plasmidial em Salmonella Muenster isoladas de suínos e do ambiente de abatedouros

Foram analisadas para a presença de plasmídios 38 amostras de Salmonella Muenster isoladas de suínos e do ambiente de abatedouros localizados no Estado do Rio de Janeiro, no período de março de 1991 a fevereiro de 1992. A seleção das amostras teve como orientação o perfil apresentado frente aos antimicrobianos estreptomicina, tetraciclina, sulfonamida e sulfametoxazol-trimetoprim, sendo analisadas 13 cepas resistentes a um ou mais antimicrobianos, 18 com grau intermediário e sete sensíveis. Plasmidios variando em tamanho de 1,2 Kb a 42 Kb foram detectados em 37 (97,36%) das 38 amostras, correspondendo a 11 perfis distintos (P1 a P11), variando em número de 1 a 6 plasmidios por modelo. O número e diversidade de plasmidios foi maior que os marcos de resistência por cepa. O plasmidio de 2,85 Kb foi o mais freqüente, estando presente em 83,78% das 37 cepas; somente o de 7,95 Kb foi detectado nos dois abatedouros. Não houve paralelismo entre padrão de resistência e perfil plasmidial, onde um mesmo antibiotipo foi encontrado em vários perfis plasmidiais. Os resultados na presente investigação permitiram concluir que a caracterização de plasmidios constituiu-se uma ferramenta útil e simples no rastreamento epidemiológico deste sorovar

Antimicrobial resistance and R-plasmid in Salmonella spp from swine and abattoir environments

Salmonella serovars isolated from swine are of particular interest not only because of the pathogenic potential for this animal species, but also due to its relevance with regard to public health. On basis of the profile of resistance to antimicrobials, 13 Salmonella strains were selected which belonged to the serovars Muenster (7), Derby (4), Typhimurium (1), and Braenderup (1). They were isolated from healthy swine as well as from the abattoir environment in the state of Rio de Janeiro. All strains of Salmonella were subjected to bacterial conjugation, and the E. coli K12 Nal r Lac+ F standard strain was used as receptor, with the purpose to verify the ability to transfer the resistance marks. Gene transfer phenomenon was detected in seven strains, and except SalmonellaTyphimurium which transconjugated to Sm, Tc and Su, the remaining ones were characterized by transferring mark Su only. By plasmidial analysis of strains used and their respective transconjugants, 63 Kb plasmid was found, which was probably related to S. Typhimurium resistance