Molecular characterization of Pasteurella multocida isolates from swine lungs by Randomly Amplified Polymorphic DNA

• Published in: Ciência rural 2016-01, Vol.46 (1), p.119-125
• Main Authors: Chitarra, Cristiane Silva, Silva, Mayara Inácio Vincenzi da, Brandão, Laila Natasha Santos, Kagueyama, Francielle Cristina, Candido, Stefhano Luis, Rosa, Janaina Marcela Assunção, Nakazato, Luciano, Dutra, Valéria
• Format: Article
• Language: English
• Subjects: Abattoirs; AGRONOMY; Amplification; Animals; Antimicrobial agents; Atrophic rhinitis; Bronchopneumonia; Deoxyribonucleic acid; Disease; DNA; DNA polymerase; Economic impact; Fragments; Genes; Genetic diversity; Hogs; Laboratories; Lesions; Livestock; Lungs; Molecular weight; Pasteurella; Pasteurella multocida; pasteurelose pulmonar; Phenols; Pneumonia; Polymerase chain reaction; RAPD; Respiratory diseases; Rhinitis; suíno; Swine; Vaccines
• ISSN: 1678-4596; 0103-8478; 1678-4596
• DOI: 10.1590/0103-8478cr20150153

Swine respiratory diseases such as atrophic rhinitis and bronchopneumonia caused by Pasteurella (P.) multocida cause important economic losses to the modern swine industry. The purpose of this study was to characterize P. multocida strains isolated from swine lungs by RAPD (Randomly Amplified Polymorphic DNA) to demonstrate their genetic diversity. Ninety-four samples of fragments from lungs with pneumonia and sixty one samples without pneumonia were collected in slaughterhouses in Mato Grosso during the period from December 2009 to March 2010. Clinical cases in 2012 and 2013 were also included in this study. Among the lung fragments with macroscopic lesions, without macroscopic lesions and clinical samples, 40.42%, 4.49% and 100% were positive for P. multocida, respectively. Bacterial identification culturing was confirmed by PCR (polymerase chain reaction) by means of the amplification of the gene kmt1. RAPD technique was performed for 46 isolates, and in every isolate, a total of 7 to 11 amplification bands were detected, composed of 8 clusters based on genetic similarity. Thus, treatment, control and preventive measures should consider the genetic diversity of P. multocida populations in swine herds in order to improve the development of new protocols to produce antimicrobials and vaccines. RESUMO: As doenças respiratórias suínas como a rinite atrófica e broncopneumonia, associada a Pasteurella (P.) multocida causam importantes perdas econômicas na suinocultura moderna. O objetivo deste trabalho foi caracterizar isolados de P. multocida de pulmão suíno através do Randomly Amplified Polymorphic DNA (RAPD) para demonstrar a diversidade genética. Noventa e quatro amostras de fragmentos de pulmões com lesões de pneumonia e sessenta e uma amostras sem lesão foram coletadas em frigoríficos no Estado do Mato Grosso, durante o período de dezembro de 2009 a março de 2010. Amostra de casos clínicos ocorridos em 2012 e 2013 também foram inlcuídos. Amostras de pulmões com lesões macroscópicas, sem lesões macroscópicas e amostras clínicas apresentaram presença de 40,42%, 4,49% e 100% de isolamento para P. multocida, respectivamente. Os isolados foram todos confirmados através da PCR (Polymerase Chain Reaction) pela amplificação do gene kmt1. A técnica de RAPD foi realizada em 46 amostras e em cada isolado foi detectado 7 a 11 bandas, que foram agrupadas em 8 grupos baseados em suas similaridades genéticas. Dessa forma, tratamento, controle e medidas preventivas dev