Release of free DNA by membrane-impaired bacterial aerosols due to aerosolization and air sampling.
Authors
Zhen-H; Han-T; Fennell-DE; Mainelis-G
Source
Appl Environ Microbiol 2013 Dec; 79(24):7780-7789
Abstract
We report here that stress experienced by bacteria due to aerosolization and air sampling can result in severe membrane impairment, leading to the release of DNA as free molecules. Escherichia coli and Bacillus atrophaeus bacteria were aerosolized and then either collected directly into liquid or collected using other collection media and then transferred into liquid. The amount of DNA released was quantified as the cell membrane damage index (ID), i.e., the number of 16S rRNA gene copies in the supernatant liquid relative to the total number in the bioaerosol sample. During aerosolization by a Collison nebulizer, the ID of E. coli and B. atrophaeus in the nebulizer suspension gradually increased during 60 min of continuous aerosolization. We found that the ID of bacteria during aerosolization was statistically significantly affected by the material of the Collison jar (glass > polycarbonate; P < 0.001) and by the bacterial species (E. coli > B. atrophaeus; P < 0.001). When E. coli was collected for 5 min by filtration, impaction, and impingement, its ID values were within the following ranges: 0.051 to 0.085, 0.16 to 0.37, and 0.068 to 0.23, respectively; when it was collected by electrostatic precipitation, the ID values (0.011 to 0.034) were significantly lower (P < 0.05) than those with other sampling methods. Air samples collected inside an equine facility for 2 h by filtration and impingement exhibited ID values in the range of 0.30 to 0.54. The data indicate that the amount of cell damage during bioaerosol sampling and the resulting release of DNA can be substantial and that this should be taken into account when analyzing bioaerosol samples.
Keywords
Bacteria; Aerosols; Microorganisms; Airborne-particles; Air-samples; Sampling-methods; Air-sampling-equipment; Air-sampling-techniques; Microbiology; DNA-adducts; Free-radicals; Cellular-structures; Cell-damage; Membrane-dysfunction; Recombinant-DNA; Genes; Filtration; Impingers; Electrostatic-precipitation; Physiological-factors; Analytical-instruments; Air-quality
Contact
Gediminas Mainelis, Rutgers University, Department of Environmental Sciences, New Brunswick, NJ, USA
Document Type
Journal Article
Email Address
mainelis@envsci.rutgers.edu
Identifying No.
Grant-Number-R01-OH-009783
Source Name
Applied and Environmental Microbiology
Performing Organization
Rutgers University, New Brunswick, New Jersey
