Application of flow cytometry for the assessment of preservation and recovery efficiency of bioaerosol samplers spiked with Pantoea agglomerans

Exposure assessment of biological aerosols requires trade-offs between efficient sampling of airborne microorganisms as either particles or viable units. The main objective of this work was to characterize aspects of bioaerosol measurement efficiency. A known concentration of the vegetative bacteria Pantoea agglomerans was spiked onto different samplers (AGI-30, BioSampler, and membrane filters) and then run for increasing time periods using HEPA filtered air. Measurement efficiency was evaluated based on total, viable, and culturable counts. Total and viable counts were determined by flow-cytometry (FCM); culturable counts were evaluated by standard plating. FCM as a method for assaying viability showed excellent agreement with known proportions of live/dead organisms (slope = 0.82, R-2 = 0.99). P. agglomerans recoveries (total, viable, and culturable) in order of best sampler performance included the BioSampler (75%, 52%, and 50%), filtration (50%, 13%, and 2%), and the AGI-30 (< 30%, 15%, and 5%). The difference between viability and culturability provided an indication of viable but nonculturable (VBNC) cells. VBNC efficiency for sampling by filter, AGI-30, and BioSampler was 80%, 50%, and 100%, respectively. This research helps characterize recovery, survival, and culturability efficiencies while sampling environmentally sensitive airborne bacteria for purposes of exposure assessment, epidemiologic studies, and homeland security.