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Experimental respiratory infection with Bordetella pertussis in mice: comparison of two methods
S A Halperin1, S A Heifetz, A Kasina
1Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia.
Abstract:
The mouse respiratory model is being used increasingly to study the pathogenesis and immunology of Bordetella pertussis infection. Two methods of inoculation, aerosol and intranasal, are routinely used to establish the infection. We compared the two methods of inoculation for reproducibility of infection using quantitative lung cultures and distribution of infection with [35S] methionine labeled bacteria and pulmonary histopathology. Ability to produce a respiratory infection intranasally was related to the inoculum volume; a minimum of 20 microliters was required although considerable variability remained. Lung bacterial counts in identically inoculated mice varied 1,000 fold following intranasal inoculation compared to only 5 fold following aerosol inoculation. Distribution of pulmonary 35S-labeled bacteria varied widely (right lung, 43-84%; left lung 16-57%) following intranasal in comparison to aerosol inoculation (right, 60-68%; left 32-40%). Finally, intranasal inoculation produced a scant, patchy, bronchopneumonia whereas diffuse pathology involving all pulmonary segments was seen following aerosol infection. Due to the superior reproducibility and predictable distribution of infection and pathology, aerosol inoculation is the method of choice for establishing the mouse model of pertussis respiratory infection.
Insights
For studying Bordetella pertussis, aerosol inoculation in mice offers superior reproducibility and predictable infection distribution compared to intranasal methods. This makes aerosol the preferred choice for establishing the mouse respiratory infection model.
Area of Science:
- Infectious Disease Research
- Immunology
- Microbiology
Background:
- The mouse respiratory model is crucial for understanding Bordetella pertussis pathogenesis and immunology.
- Aerosol and intranasal inoculation are common methods for establishing respiratory infections in mice.
Purpose of the Study:
- To compare the reproducibility and infection characteristics of aerosol versus intranasal inoculation methods for Bordetella pertussis in mice.
- To determine the optimal inoculation method for establishing a reliable mouse model of pertussis respiratory infection.
Main Methods:
- Comparison of aerosol and intranasal inoculation techniques in mice.
- Quantitative lung cultures to assess bacterial load.
- Use of [35S] methionine labeled bacteria to track infection distribution.
- Pulmonary histopathology to evaluate disease severity and spread.
Main Results:
- Intranasal inoculation required a minimum of 20 microliters, with significant variability in infection reproducibility (1,000-fold variation in lung bacterial counts).
- Aerosol inoculation demonstrated superior reproducibility (only 5-fold variation) and a more predictable distribution of infection.
- Histopathology revealed patchy bronchopneumonia with intranasal inoculation versus diffuse pathology throughout the lungs with aerosol inoculation.
Conclusions:
- Aerosol inoculation provides significantly better reproducibility and more predictable infection distribution and pathology compared to intranasal inoculation.
- Aerosol inoculation is the recommended method for establishing the mouse model of Bordetella pertussis respiratory infection due to its superior performance.