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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2018.tde-08022018-152121
Document
Author
Full name
Nadia Catalina Alfonso Vargas
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2017
Supervisor
Committee
Schenberg, Ana Clara Guerrini (President)
Ferreira, Rita de Cassia Cafe
Maldonado, Gabriel Padilla
Sato, Maria Inês Zanoli
Simionato, Maria Regina Lorenzetti
Title in Portuguese
Caracterização, detecção  e quantificação de Vibrio cholerae em amostras de água.
Keywords in Portuguese
Galleria mellonella
Vibrio cholerae
Análise de sequências multilocus
Antibiograma
Genes associados à virulência
PCR em tempo real
Abstract in Portuguese
Vibrio cholerae é uma bactéria autóctone em ecossistemas aquáticos, os fatores responsáveis pela virulência podem contribuir com a patogenicidade, influenciados por fatores genéticos e ambientais. Considerando a importância de conhecer e monitorar o V. cholerae, o estudo pretende caracterizar isolados da especie e padronizar uma metodologia para detecção em amostras de água. Os isolados foram avaliados por metodologias clássicas e moleculares, para confirmar espécie. Também, foi avaliada a presença de genes de virulência, susceptibilidade aos antibióticos e resposta em modelo invertebrado. Tres marcadores moleculares foram avaliados por PCR quantitativa. Observou-se que setenta dos isolados pertenciam a espécie V. cholerae e mostraram variação na prevalência dos genes de virulência e ao perfil de suscetibilidade ao antibióticos. Mostrou uma influencia da temperatura e concentração do inoculo no modelo invertebrado. Os marcadores moleculares selecionados mostraram a viabilidade da metodologia proposta neste estudo pela alta especificidade e sensibilidade.
Title in English
Characterization, detection and quantification of Vibrio cholerae in water samples.
Keywords in English
Galleria mellonella
Vibrio cholerae
Antibiogram
Factors associated with virulence
Multilocus sequence analysis
Real time PCR
Abstract in English
Vibrio cholerae is an autochthonous bacterium in aquatic ecosystems, factors responsible for virulence may contribute to pathogenicity, influenced by genetic and environmental factors. Considering the importance of knowing and monitoring V. cholerae, the study pretend to characterize selected isolates and to standardize a methodology for detection in water samples. The isolates were evaluated by classical and molecular methodologies to confirm species. Also, the presence of factors associated with virulence, antibiotics susceptibility and response in invertebrate model were evaluated. Three molecular markers were evaluated by quantitative PCR. It was observed that seventy of the isolates belonged to the V. cholerae species and showed a variation in the prevalence of the virulence genes and the antibiotic susceptibility profile. Also, showed an influence of the inoculum temperature and concentration on the invertebrate model. The selected molecular markers showed the viability of the methodology proposed in this study for the high specificity and sensitivity.
 
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Publishing Date
2018-02-08
 
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