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Home / Equine Research Bank / Appl Environ Microbiol / Release of free DNA by membrane-impaired bacterial aerosols ...
Applied and environmental microbiology2013; 79(24); 7780-7789; doi: 10.1128/AEM.02859-13

Release of free DNA by membrane-impaired bacterial aerosols due to aerosolization and air sampling.

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.
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Publication Date: 2013-10-04 PubMed ID: 24096426PubMed Central: PMC3837826DOI: 10.1128/AEM.02859-13Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article explores how stress on bacteria due to aerosolization and air sampling can lead to considerable membrane damage, causing the bacteria to release DNA as free molecules. This has crucial implications for the analysis of bioaerosol samples.

Understanding the Process

  • The researchers conducted experiments on Escherichia coli and Bacillus atrophaeus bacteria which were aerosolized and then collected either directly into liquid or through other collection media and then into liquid.
  • The DNA amount released as a result of membrane impairment was calculated as the cell membrane Damage Index (ID). The Damage Index represents the ratio of the number of 16S rRNA gene copies present in the supernatant liquid to the total number in the bioaerosol sample. This method offered a quantifiable way of measuring the amount of DNA released due to cell damage

Environmental Influences on Bacterial DNA Release

  • The study found that during aerosolization by a Collison nebulizer, the Damage Index of E. coli and B. atrophaeus in the nebulizer suspension gradually rose over a period of 60 minutes of continuous aerosolization.
  • The study also revealed that the material of the Collison jar (glass vs polycarbonate), and the bacterial species (E. coli vs B. atrophaeus) significantly affected the Damage Index of bacteria during aerosolization.

Impact of Sampling Methods on Bacterial DNA Release

  • It was discovered that the method of collection could significantly influence the Damage Index. When E. coli was collected for 5 minutes by filtration, impaction, and impingement, its Damage Index values varied considerably. However, when it was collected by electrostatic precipitation, the Damage Index values were significantly lower than those with other sampling methods.
  • This pattern was also seen in air samples collected inside an equine facility for 2 hours by filtration and impingement that exhibited Damage Index values in a certain range.

Considerations for Future Analysis of Bioaerosol Samples

  • This study suggests that the amount of cell damage during bioaerosol sampling and the consequent release of DNA can be substantial.
  • This is an important consideration for future bioaerosol sample analysis, as released DNA could potentially alter the interpretation of such samples. Therefore, studying the conditions and factors that increase or decrease DNA release through cell damage could greatly enhance the accuracy of bioaerosol analysis.

Cite This Article

APA
Zhen H, Han T, Fennell DE, Mainelis G. (2013). Release of free DNA by membrane-impaired bacterial aerosols due to aerosolization and air sampling. Appl Environ Microbiol, 79(24), 7780-7789. https://doi.org/10.1128/AEM.02859-13

Publication

Applied and environmental microbiology
ISSN: 1098-5336
NlmUniqueID: 7605801
Country: United States
Language: English
Volume: 79
Issue: 24
Pages: 7780-7789

Researcher Affiliations

Zhen, Huajun
  • Rutgers University, Department of Environmental Sciences, New Brunswick, New Jersey, USA.
Han, Taewon
    Fennell, Donna E
      Mainelis, Gediminas

        MeSH Terms

        • Aerosols
        • Air Microbiology
        • Animals
        • Bacillus / physiology
        • Cell Membrane / physiology
        • DNA, Bacterial / isolation & purification
        • Escherichia coli / physiology
        • Horses
        • Housing, Animal
        • Specimen Handling / methods
        • Stress, Mechanical

        Grant Funding

        • R01 OH009783 / NIOSH CDC HHS
        • R01OH009783 / ACL HHS
        • R01-OH009783 / NIOSH CDC HHS

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