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. 2002 Sep;68(9):4646-9.
doi: 10.1128/AEM.68.9.4646-4649.2002.

Infection of mice with aerosolized Mycobacterium tuberculosis: use of a nose-only apparatus for delivery of low doses of inocula and design of an ultrasafe facility

J Reid Schwebach  1 , Bing Chen, Aharona Glatman-Freedman, Arturo Casadevall, John D McKinney, John L Harb, Patrick J McGuire, W Emmett Barkley, Barry R Bloom, William R Jacobs Jr
Affiliations

Infection of mice with aerosolized Mycobacterium tuberculosis: use of a nose-only apparatus for delivery of low doses of inocula and design of an ultrasafe facility

J Reid Schwebach et al. Appl Environ Microbiol. 2002 Sep.

Abstract

Aerosolized delivery of virulent or hypervirulent Mycobacterium tuberculosis requires careful consideration of methodology and safety. To maximize safety, we installed a nose-only aerosol apparatus that can reproducibly deliver a low dose (<100 CFU per mouse) of M. tuberculosis in a carefully designed biohazard facility.

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Figures

FIG. 1.
FIG. 1.
Facility floorplan. The BSL-3 facility contains an anteroom (A), a work area complete with multiple biosafety cabinets (B), an animal room (C), and an aerosol room (D). An autoclave joins the aerosol room with the anteroom (D), and a shower room (E) is available for emergency exit. Dimensions of the facility and appliances are indicated in feet. Airflow is indicated by arrows. The aerosol cabinet is also indicated (F).
FIG. 2.
FIG. 2.
Facility and aerosol apparatus photographs. The aerosol cabinet (F) is joined to a biohazard cabinet in the animal room (A) via a sealed, interconnecting passageway (C) that connects the two cabinets between the two rooms. Glove openings in the glove box are shown as dotted lines (J). Airflow in the rooms is indicated above the doorways (ball is pushed by air movement) (B). The control panel of the aerosol apparatus is shown (I), and the nebulizer is illustrated (H). The animal chamber is shown (E), and an AGI (G) and an animal stanchion containing a mouse (D) are illustrated. Mice (L) are contained in stanchions (K and M) that connect to the animal chamber. The nose of each mouse rests inside an aperture of the stanchion (M), allowing for nose-only aerosol exposure. The airflow direction within the aerosol apparatus is indicated by arrows.
FIG. 3.
FIG. 3.
Delivery of M. smegmatis to mice. Total CFU in the lung versus CFU per milliliter in the nebulizer as a result of the aerosolization procedure is shown. Each value represents the average for three animals. Error bars show the standard deviations of the means.
FIG. 4.
FIG. 4.
Delivery of M. tuberculosis Erdman to mice. Total CFU in the lung versus CFU per milliliter in the nebulizer as a result of the aerosolization procedure is shown. Each value represents the average for three animals. Error bars show the standard deviations of the means.
See this image and copyright information in PMC

References

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