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Nanoscale aerovirology: An efficient yet simple method to analyze the viral distribution of single bioaerosols

ABSTRACT The aerosolization mechanisms of viruses are poorly known, because of the challenges related to their sampling and observation. For example, single particle studies are needed to improve our understanding of bioaerosol enrichment processes. Such studies would help to develop models of airborne disease propagation. We propose a novel approach to study viral aerosols in single particles using a combination of fluorescence and transmission electron microscopy (TEM). This method allows for rapid analysis of labeled bacteriophages aerosolized and captured on a black membrane filter. It also requires performing image analyses on fluorescent spots. TEM is necessary to determine a single bacteriophage dimensions. Thus, the clustering of bacteriophage PP01 in a single aerosol particle was investigated and found to give a comparable number of virions to what was observed with TEM. The impact of the GFP (green fluorescent protein) in the head of PP01 virion compared to wild type (WT) PP01 was also tested by comparing the clustering of similar bioaerosol sizes generated by the aerosolization of PP01 WT, PP01-GFP, and PP01-GFP labeled with syto-red dye. Surprisingly, the PP01 WT bioaerosols were enriched compared to the PP01-GFP ones (64.9 ± 17.5% more). PP01-GFPs were also found to be more numerous compared to those produced by PP01-GFP labeled with syto-red dye (28.9 ± 16.9% more). The aerosolization process might be dependent on the electrochemical properties of the viruses and the environment. Changes of this nature could affect the mechanism of the aerosol formation in natural forming aerosols as demonstrated in this study for artificially generated aerosols. © 2016 American Association for Aerosol Research