A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis

Pulmonary melioidosis, a disease manifestation caused by the bacterium Burkholderia pseudomallei, has been studied using aerosols or intranasal inoculation in small animal models. Both have inherent disadvantages which may not accurately model primary pulmonary melioidosis in humans. Intratracheal...

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Main Authors: David Aaron Revelli, Julie A Boylan, Frank C Gherardini
Format: Article
Language:English
Published: Frontiers Media S.A. 2012-12-01
Series:Frontiers in Cellular and Infection Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/Journal/10.3389/fcimb.2012.00164/full
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author David Aaron Revelli
Julie A Boylan
Frank C Gherardini
author_facet David Aaron Revelli
Julie A Boylan
Frank C Gherardini
author_sort David Aaron Revelli
collection DOAJ
description Pulmonary melioidosis, a disease manifestation caused by the bacterium Burkholderia pseudomallei, has been studied using aerosols or intranasal inoculation in small animal models. Both have inherent disadvantages which may not accurately model primary pulmonary melioidosis in humans. Intratracheal inoculation by direct visualization of the tracheal opening offers an alternative technique for infection that overcomes the disadvantages of aerosol and intranasal challenge. In this study, we describe a method which requires relatively inexpensive equipment, little training, and is compliant with the operational constraints of a BSL3 laboratory. Results obtained using trypan demonstrated that an inoculum can be accurately delivered into the lungs of mice within a biosafety cabinet. Whole body imaging and histopathology confirmed that mice inoculated intratracheally with B. pseudomallei develop the primary focus of infection in the lungs, and not the nasal passages which can lead to invasion of the central nervous system and potential neurologic complications. Further, based on colony counts and bioluminescent imaging, dissemination to secondary organs occurred as expected. Taken together, this intratracheal method of inoculation fulfills four goals: (1) to accurately deliver B. pseudomallei into the lungs of the animal model, (2) to avoid potentially confounding complications due to primary infections at sites other than the lung, (3) to maintain normal organ dissemination, and (4) to be BSL3 compliant.
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spelling doaj.art-8ed75a26abfd4d9080acdc72fcd689c62022-12-22T01:30:41ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882012-12-01210.3389/fcimb.2012.0016440191A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary MelioidosisDavid Aaron Revelli0Julie A Boylan1Frank C Gherardini2National Institutes of Allergy and Infectious Disease, National Institutes of HealthNational Institutes of Allergy and Infectious Disease, National Institutes of HealthNational Institutes of Allergy and Infectious Disease, National Institutes of HealthPulmonary melioidosis, a disease manifestation caused by the bacterium Burkholderia pseudomallei, has been studied using aerosols or intranasal inoculation in small animal models. Both have inherent disadvantages which may not accurately model primary pulmonary melioidosis in humans. Intratracheal inoculation by direct visualization of the tracheal opening offers an alternative technique for infection that overcomes the disadvantages of aerosol and intranasal challenge. In this study, we describe a method which requires relatively inexpensive equipment, little training, and is compliant with the operational constraints of a BSL3 laboratory. Results obtained using trypan demonstrated that an inoculum can be accurately delivered into the lungs of mice within a biosafety cabinet. Whole body imaging and histopathology confirmed that mice inoculated intratracheally with B. pseudomallei develop the primary focus of infection in the lungs, and not the nasal passages which can lead to invasion of the central nervous system and potential neurologic complications. Further, based on colony counts and bioluminescent imaging, dissemination to secondary organs occurred as expected. Taken together, this intratracheal method of inoculation fulfills four goals: (1) to accurately deliver B. pseudomallei into the lungs of the animal model, (2) to avoid potentially confounding complications due to primary infections at sites other than the lung, (3) to maintain normal organ dissemination, and (4) to be BSL3 compliant.http://journal.frontiersin.org/Journal/10.3389/fcimb.2012.00164/fullBurkholderia pseudomalleiMelioidosispulmonary melioidosisintratracheal inoculationnoninvasive inoculation method
spellingShingle David Aaron Revelli
Julie A Boylan
Frank C Gherardini
A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis
Frontiers in Cellular and Infection Microbiology
Burkholderia pseudomallei
Melioidosis
pulmonary melioidosis
intratracheal inoculation
noninvasive inoculation method
title A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis
title_full A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis
title_fullStr A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis
title_full_unstemmed A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis
title_short A Noninvasive Intratracheal Inoculation Method for the Study of Pulmonary Melioidosis
title_sort noninvasive intratracheal inoculation method for the study of pulmonary melioidosis
topic Burkholderia pseudomallei
Melioidosis
pulmonary melioidosis
intratracheal inoculation
noninvasive inoculation method
url http://journal.frontiersin.org/Journal/10.3389/fcimb.2012.00164/full
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