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Doctoral Thesis
DOI
https://doi.org/10.11606/T.10.2016.tde-21062016-124945
Document
Author
Full name
Givago Faria Ribeiro da Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Moreno, Andrea Micke (President)
Andrade, André Furugen Cesar de
Calderaro, Franco Ferraro
Knöbl, Terezinha
Oliveira, Luís Guilherme de
Title in Portuguese
Caracterização fenotípica e molecular de estirpes de Haemophilus parasuis isoladas de suínos da região Centro-sul do Brasil
Keywords in Portuguese
Haemophilus parasuis
MLST
Resistencia
Sorotipagem
Virulência
Abstract in Portuguese
Haemophilus parasuis é o agente etiológico da Doença de Glässer, que causa artrite, pneumonia, meningite e poliserosite em suínos e tem assumido grande importância na suinocultura moderna, uma vez que sua ocorrência tem aumentado significativamente nos últimos anos em rebanhos afetados pelo circovirus suíno tipo 2. No presente estudo foram avaliadas 117 amostras de H. parasuis isoladas dentre os anos de 2009 a 2014, isoladas de suínos de diferentes estados do da região Centro-Sul do Brasil. As estirpes foram submetidas à sorotipificação, confirmação do gênero/espécie pela PCR, o perfil de resistência a antimicrobianos foi avaliado através da determinação da concentração inibitória mínima (CIM), foi realizada a caracterização genotípica das amostras por eletroforese em gel de campo pulsado (PFGE) e por sequenciamento de múltiplos sítios (multilocus sequence typing - MLST) e a presença de genes de virulência vtaA foi analisada. Os sorotipos mais frequentes foram: 4 (21,3%), seguido do 5 (12,9%), do 13 (9,4%), do 14 (7,7%) e do sorotipo 1 (1,7%), e em alguns casos mais de um sorotipo foi identificado na mesma granja e até no mesmo animal, resultado este parecido ao encontrado no restante do mundo. Em todas as amostras o gene vtaA estava presente, para alguns antibióticos os índices de resistência foram elevados, como para tilosina (98,29%), danofloxacina (95,72%), sulfadimetoxina (88,03%), penicilina (77,7%) e a multirrestencia atingiu o índice alarmente de 93,16% das estirpes. Foram identificados 67 perfis diferentes no PFGE e das 9 amostras analisadas pelo MLST foram identificados novos STs, até então, não descritos mundialmente. Quando os novos STs foram comparados com os previamente descritos, estas se dispersaram entre as descritas em diferentes países. Neste estudo foi possível observar que as estirpes de H. parasuis brasileiras possuem alta variabilidade, tanto nos sorotipos, perfis de resistência, análises genômicas de PFGE e MLST
Title in English
Phenotypic and molecular characterization of Haemophilus parasuis strains isolated from pigs in the Center- South of Brazil
Keywords in English
Haemophilus parasuis
MLST
Resistance
Serotyping
Virulence
Abstract in English
Haemophilus parasuis is the etiological agent of Glässer disease that causes arthritis, pneumonia, meningitis and polyserositis in pigs and has assumed great importance in modern swine production, since its occurrence has increased significantly in recent years in herds affected by porcine circovirus type 2. In the present study 117 strains of H. parasuis isolated between 2009 to 2014 were utilized, isolated from pigs of south center of Brazil. The strains were serotyping, confirmed genus/species by PCR, the antimicrobial resistance profile was evaluated determining the minimum inhibitory concentration (MIC) and strains were genotypically characterized by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and presence of vtA virulence gene. The major serotypes identified were 4 (21.3%), 5 (12.9%), 13 (9.4%), 14 (7.7%) and at last the 1 (1.7%), In some cases more than one serotype was identified in the same farm and in the same animal, this results were identified in others parts of the world. All samples had vtA gene. The resistance for some antibiotics was high for tylosin (98.29%), danofloxacin (95.72%) sulfadimetoxin (88.03%), and penicillin (77.7%). Multidrug resistance rates reached 93.16% of the samples. A total of 67 different profiles were identified in PFGE and nine samples were analyzed by MLST. All nine strains tested were identified as new STs. When these strains were compared with MLST database, they were dispersed among the strains from other countries. In this study, it was clear that the Brazilian H. parasuis strains are highly variable considering serotypes, resistance profiles, genomic analysis of PFGE and MLST
 
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Publishing Date
2016-09-05
 
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