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Master's Dissertation
Document
Author
Full name
Mirela Caroline Vilela de Oliveira
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Knöbl, Terezinha (President)
Menão, Marcia Cristina
Moreno, Andrea Micke
Title in Portuguese
Análise filogenética de Salmonella Typhimurium e Escherichia coli diarreiogênica isoladas de pombos (Columba livia) que realizam o forrageamento em recintos de zoológicos: implicações zoonóticas
Keywords in Portuguese
Columba livia
EPEC
Salmoneloses
STEC
Zoonoses
Abstract in Portuguese
Pombos (Columba livia) são aves exóticas, sinantrópicas, reservatórios de algumas doenças de caráter zoonótico. O presente trabalho teve por objetivo caracterizar genotipicamente isolados de Salmonella spp. e Escherichia coli diarreiogênicas de pombos domésticos. Foram coletados suabes de 118 pombos provenientes de dois zoológicos de São Paulo. Para a identificação de Salmonella spp, as amostras foram submetidas ao pré-enriquecimento em água peptonada 0,1% e incubadas a 37º C por 24-48 horas, seguida do enriquecimento seletivo em caldo de tetrationato de sódio, com incubação a 37º C por 24-48h e plaqueamento em meio seletivo XLT4, a 37º C por 24-48h. Para o isolamento de E. coli, as amostras foram semeadas em ágar MacConkey, com incubação a 37º C por 24 h. As colônias suspeitas foram identificadas por série bioquímica. A técnica de PCR foi utilizada para a confirmação do gênero Salmonella spp, identificação dos sorovares de S. Enteritidis e S. Typhimurium e para a determinação dos fatores de virulência de Escherichia coli. Dentre os 118 animais analisados, 4,23% (5/118) foram positivos para Salmonella spp, sendo todos do sorovar S. Typhimurium. Foram identificadas estirpes diarreiogênicas de E. coli, positivas para os fatores de virulência eae (8/118), bfp (4/118) e stx2f (3/118). Na análise filogenética pela técnica de AFLP, as estirpes de E. coli diarreiogênicas apresentaram uma diversidade genética, mas as estirpes de S. Typhimurium foram consideradas clonais. Os dados deste trabalho destacam o risco sanitário relacionado ao forrageamento da espécie Columba livia em zoológicos, por meio da confirmação da presença de Salmonella Typhimurium, EPEC típica, EPEC atípica e STEC nas excretas destas aves
Title in English
Phylogenetic analysis of Salmonella Typhimurium and diarrheagenic Escherichia coli isolated a pigeons (Columba livia) that perform foraging in zoos grounds: zoonotic implications
Keywords in English
Columba livia
EPEC
Salmonellosis
STEC
Zoonoses
Abstract in English
Pigeons (Columba livia) are exotic birds, synanthropic and reservoirs of some zoonotic diseases. The aim of this study was to characterize genotypically isolates of Salmonella spp. and diarrheagenic strains of Escherichia coli isolated from domestic pigeons. Fecal swabs of 118 pigeons were collected from two zoos in São Paulo. The samples were subjected to a pre-enrichment step being placed in peptone water solution of 0.1% and incubated for 24-48 h at 37º C. Thereafter it was performed selective enrichment step where the samples were transferred in sodium tetrathionate broth with incubation for 24-48 h at 37º C, followed by plating on selective medium XLT4 incubation for 24-48h at 37º C for isolation and identification of Salmonella spp. The samples were also placed in BHI broth for a period of 24 h and then plated on MacConkey medium with incubation for 24h at 37º C for isolation of E. coli. Subsequent microbiological step, the isolates were subjected to DNA extraction using the methodology described by Boom for PCR. Polymerase chain reaction (PCR) was employed for identification of the Salmonella serovar Enteritidis and Typhimurium and virulence factors of E. coli. Of the 118 animals tested, 4.23% (5/118) were positive for Salmonella spp., all characterized as being serovar S. Typhimurium. Some E. coli strains (8/118) were positive for virulence factors: eae, 3,38% (4/118), bfp and 2.54% (3/118) to stx2f. In phylogenetic analysis by AFLP, strains of diarrheagenic E. coli showed a genetic diversity, but S. Typhimurium presented a clonal relationship. Data from this study highlight the health risk related to the foraging of Columba livia in zoos, because they can be reservoirs of Salmonella Typhimurium, typical and atypical EPEC and STEC
 
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Publishing Date
2017-01-26
 
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