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Doctoral Thesis
DOI
10.11606/T.42.2009.tde-29102009-091226
Document
Author
Full name
Mauricio Yamaguti
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Timenetsky, Jorge (President)
Castro, Antonio Fernando Pestana de
Ferreira, Antônio José Piantino
Moreno, Andrea Micke
Nascimento, Elmiro Rosendo do
Title in Portuguese
Isolamento de micoplasma de suínos com problemas respiratórios e tipificação dos isolados pela PFGE e seqüenciamento do gene 16S rRNA.
Keywords in Portuguese
Mycoplasma hyopneumoniae
Mycoplasma hyorhinis
Gene 16S rRNA
Microbiologia
Multiplex-PCR
PFGE
Seqüenciamento genético
Suínos
Abstract in Portuguese
As doenças respiratórias são responsáveis, na suinocultura, por grandes perdas econômicas e entre os agentes destacam-se os micoplasmas. Foram coletadas 126 amostras de muco nasal/tonsilar, e fragmentos de 78 pulmões, 2 de traquéia e 2 de tonsila. No isolamento foi utilizado o meio FRIIS modificado, na identificação, a Multiplex-PCR e na tipificação, a PFGE e sequenciamento do gene 16S rRNA. Foram obtidos 59 isolados identificados como M. hyopneumoniae (1,70%), M. flocculare (3,40%) e M. hyorhinis (94,90%). A PFGE dos isolados de M. hyorhinis, resultou em 10 e 9 perfis com a enzima AvaI e XhoI, respectivamente. O sequenciamento do gene 16S rRNA dos isolados M. hyorhinis apresentaram baixo polimorfismo quando comparados entre si e com a cepa de referência. A sequência do gene 16S rRNA do isolado de M. hyopneumoniae, quando comparada a seqüência da cepa J e os isolados 7448 e 232, resultaram em polimorfismo. O M. hyopneumoniae continua sendo o mais difícil de isolar. Os dendrogramas obtidos da PFGE resultaram em grande heterogeneidade entre os isolados de M. hyorhinis. O seqüenciamento do gene 16S rRNA permitiu o estudo de variabilidade interespecífica e intraespecífica dos isolados de micoplasmas.
Title in English
Isolation of swine origin mycoplasmas from specimens with respiratory disturbances and typification of isolates by PFGE and 16S rRNA sequencing gene.
Keywords in English
16S rRNA gene
Mycoplasma hyopneumoniae
Mycoplasma hyorhinis
Genetic Sequencing
Microbiology
Multiplex-PCR
PFGE
Swines
Abstract in English
Economic losses in swine production are common due the respiratory diseases in these animals. M. hyopneumoniae and M. hyorhinis are the most frequent microbial agents. The aim of this study was recover this isolates in FRIIS medium, indentify them by Multiplex PCR and detect their genotypic variations by PFGE and sequencing the 16s rRNA gene. One hundred twenty six swabs from tonsil and nasal mucus of swine with respiratory disturbances were analyzed. It was included 78 lungs, two trachea and two tonsils. It was obtained 59 isolates; 1.70% of M. hyopneumoniae, 3.40% of M. flocculare and 94.90% of M. hyorhinis. The PFGE of M. hyorhinis, allowed obtain 10 profiles with enzyme AvaI and nine profiles with XhoI. A low polymorphism of gene 16sRNS was detected in M. hyorhinis isolates when compared with the type strain at the GenBank. The M. hyopneumoniae isolates resulted in polymorphisms when comparated with strain J, 7448 and 232. M. hyopneumoniae is still the most difficult to isolate. M. hyorhinis isolates of different herds showed a large heterogenicity with enzymes AvaI e XhoI. The sequencing of gene 16S rRNA allowed analyse the interespecífic and intraespecífic variations of mycoplasmas isolated.
 
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Publishing Date
2010-03-08
 
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