Sampling port for real-time analysis of bioaerosol in whole body exposure system for animal aerosol model development.
BACKGROUND
Multiple factors influence the viability of aerosolized bacteria. The delivery of aerosols is affected by chamber conditions (humidity, temperature, and pressure) and bioaerosol characteristics (particle number, particle size distribution, and viable aerosol concentration). Measurement of viable aerosol concentration and particle size is essential to optimize viability and lung delivery. The Madison chamber is widely used to expose small animals to infectious aerosols.


METHODS
A multiplex sampling port was added to the Madison chamber to measure the chamber conditions and bioaerosol characteristics. Aerosols of three pathogens (Bacillus anthracis, Yersinia pestis, and Mycobacterium tuberculosis) were generated under constant conditions and their bioaerosol characteristics were analyzed. Airborne microbes were captured using an impinger or BioSampler. The particle size distribution of airborne microbes was determined using an aerodynamic particle sizer (APS). Viable aerosol concentration, spray factor (viable aerosol concentration/inoculum concentration), and dose presented to the mouse were calculated. Dose retention efficiency and viable aerosol retention rate were calculated from the sampler titers to determine the efficiency of microbe retention in lungs of mice.


RESULTS
B. anthracis, Y. pestis, and M. tuberculosis aerosols were sampled through the port. The count mean aerodynamic sizes were 0.98, 0.77, and 0.78 μm with geometric standard deviations of 1.60, 1.90, and 2.37, and viable aerosol concentrations in the chamber were 211, 57, and 1 colony-forming unit (CFU)/mL, respectively. Based on the aerosol concentrations, the doses presented to mice for the three pathogens were 2.5e5, 2.2e4 and 464 CFU.


CONCLUSIONS
Using the multiplex sampling port we determined whether the animals were challenged with an optimum bioaerosol based on dose presented and respirable particle size.
Saini
Divey
D
Duke Human Vaccine Institute, PO Box 103020, Duke University Medical Center, Durham, North Carolina 27710, USA. divey.saini@duke.edu
Hopkins
Gregory W
GW
Chen
Ching-Ju
CJ
Seay
Sarah A
SA
Click
Eva M
EM
Lee
Sunhee
S
Hartings
Justin M
JM
Frothingham
Richard
R
D000336:Aerosols / Q000032:analysis*; D000399:Air Pressure; D000818:Animals; D001273:Atmosphere Exposure Chambers; D004867:Equipment Design; D006813:Humidity; D019570:Inhalation Exposure / Q000032:analysis*; D051379:Mice; D008810:Mice, Inbred C57BL; D050296:Microbial Viability; D023421:Models, Animal; D010316:Particle Size; D013696:Temperature
D000336:Aerosols