Release of Free DNA by Membrane-Impaired Bacterial Aerosols Due to Aerosolization and Air Sampling

Zhen, Huajun; Han, Taewon; Fennell, Donna E.; Mainelis, Gediminas

doi:10.1128/aem.02859-13

Cited by 79 publications

(78 citation statements)

References 60 publications

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“…Some fraction of bacteria might be completely damaged and not counted under a microscope thus also leading to a higher bioluminescence output per bacterium. Damage to the integrity of the bacterial membrane was also observed in our previous study of DNA release from Escherichia coli (also a gram-negative organism) after aerosolization (Zhen et al, 2013). Our previous aerosolization studies with P. fluorescens observed damage to bacterial culturability due to prolonged aerosolization (Mainelis et al, 2005).…”

Section: Resultssupporting

confidence: 64%

“…Our previous aerosolization studies with P. fluorescens observed damage to bacterial culturability due to prolonged aerosolization (Mainelis et al, 2005). Since B. atrophaeus is a gram positive bacterium with a thick outer wall, the mechanical stress of aerosolization and sampling does not result in the cell membrane rupture (Zhen et al, 2013), but apparently the stress is still sufficient to reduce RLU output per bacterium as illustrated with RR ( Collison ) values being below unity. Here again, since the polycarbonate jar is able to absorb part of the impaction energy from the bacteria, it produces lower stress and higher RR ( Collison ) compared to the glass jar.…”

Section: Resultsmentioning

confidence: 99%

“…Our earlier study showed that when bacteria are aerosolized by a Collison nebulizer, different jar material, i.e., polycarbonate vs glass, produces different effect on the integrity of bacterial cells, likely due to higher flexibility of polycarbonate compared to glass (Zhen et al, 2013). Therefore, to assess the effect of aerosolization by the Collison nebulizer on the physiological state of the test bacteria and the resulting bioluminescence signal, glass and polycarbonate jars were used and the resulting bioluminescence signals were compared..…”

Section: Methodsmentioning

confidence: 99%

“…At the same time, different bioaerosol sampling methods are known to differ in their effect on bioaerosol culturability (Zhao et al, 2011) and cell-wall integrity (Thomas et al, 2011; Zhen et al, 2013). Since ATP-based bioluminescence is based on metabolic activity of microorganisms, the strength of ATP bioluminescence signal and accuracy of bioaerosol analysis may therefore be affected by the chosen sampling method as well.…”

Section: Introductionmentioning

confidence: 99%

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Application of ATP-based bioluminescence for bioaerosol quantification: Effect of sampling method

Han

Wren

DuBois

et al. 2015

Journal of Aerosol Science

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An adenosine triphosphate (ATP)-based bioluminescence has potential to offer a quick and affordable method for quantifying bioaerosol samples. Here we report on our investigation into how different bioaerosol aerosolization parameters and sampling methods affect bioluminescence output per bacterium, and implications of that effect for bioaerosol research. Bacillus atrophaeus and Pseudomonas fluorescens bacteria were aerosolized by using a Collison nebulizer (BGI Inc., Waltham, MA) with a glass or polycarbonate jar and then collected for 15 and 60 min with: (1) Button Aerosol Sampler (SKC Inc., Eighty Four, PA) with polycarbonate, PTFE, and cellulose nitrate filters, (2) BioSampler (SKC Inc.) with 5 and 20 mL of collection liquid, and (3) our newly developed Electrostatic Precipitator with Superhydrophobic Surface (EPSS). For all aerosolization and sampling parameters we compared the ATP bioluminescence output per bacterium relative to that before aerosolization and sampling. In addition, we also determined the ATP reagent storage and preparation conditions that that do not affect the bioluminescence signal intensity. Our results show that aerosolization by a Collison nebulizer with a polycarbonate jar yields higher bioluminescence output per bacterium compared to the glass jar. Interestingly enough, the bioluminescence output by P. fluorescens increased substantially after its aerosolization compared to the fresh liquid suspension. For both test microorganisms, the bioluminescence intensity per bacterium after sampling was significantly lower than that before sampling suggesting negative effect of sampling stress on bioluminescence output. The decrease in bioluminescence intensity was more pronounces for longer sampling times and significantly and substantially depended on the sampling method. Among the investigated method, the EPSS was the least injurious for both microorganisms and sampling times. While the ATP-based bioluminescence offers a quick bioaerosol sample analysis method, this works demonstrates that the method output depends on bioaerosol generation and sampling methods, as well as reagent storage.

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Section: Resultssupporting

confidence: 64%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Application of ATP-based bioluminescence for bioaerosol quantification: Effect of sampling method

Han

Wren

DuBois

et al. 2015

Journal of Aerosol Science

Self Cite

View full text Add to dashboard Cite

show abstract

“…The prototype EPSS has also been successfully tested with microscopy, ATP and qPCR sample analysis methods (Han et al, 2010(Han et al, , 2011(Han et al, , 2015. It also preserved integrity of the collected bacteria better than an impactor or an impinger (Zhen et al, 2013). The prototype EPSS model was further developed as a single-stage electrostatic collector with more effective materials for the sampler body and electrode, and optimized ionizer configuration (EPSS, Mark II) (Han et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Design and Evaluation of the Field-Deployable Electrostatic Precipitator with Superhydrophobic Surface (FDEPSS) with High Concentration Rate

Han

Zhen

Fennell

et al. 2015

Aerosol Air Qual. Res.

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Here we report on further development of an electrostatics-based bioaerosol collector with high concentration rate. We developed a field-deployable version of the electrostatic precipitator with superhydrophobic surface (FDEPSS), which consists of two combined half-cylinder collection chambers and an integrated control box. The collector is made of a static dissipative material and each collection chamber features a 3.2 mm wide collection electrode. The round top part of each chamber contains eight carbon fiber ionizers arranged in two lines of four. The collected particles are removed by a 20 µL rolling water droplet. Sampler's components were integrated into a control box.The FDEPSS was tested with two bacterial species, Bacillus atropheus and Pseudomonas fluorescens bacteria, and one fungal spore, Penicillium chrysogenum for 10 and 60 min collection times and showed collection efficiency of ~70% at a sampling flow rate of 20 L min -1 . The use of a collecting water droplet of 20 µL per collection chamber achieved sample concentration rates approaching 0.5 × 10 6 min -1 . The FDEPSS was also tested against BioSampler (SKC Inc., Eighty Four, PA) and Button aerosol sampler (SKC Inc.) when sampling bioaerosols outdoors for 60 min. The samples were characterized based on the total airborne adenosine triphosphate (ATP) concentration, which was reported as relative luminescence units (RLU). The FDEPSS detected 5.1 × 10 5 RLU m -3 , while the BioSampler and the Button sampler showed 4.1 × 10 5 RLU m -3 and 8.7 × 10 5 RLU m -3 , respectively. Since ATP analysis can be performed with small sample volumes, and the FDEPSS captures particles into 20 µL of liquid, resulting in a high concentration rate, we show that this sampler can detect the presence of airborne microorganisms 40× faster than the BioSampler or Button aerosol sampler. This FDEPSS feature could be integrated into bioaerosol detection systems, especially where concentrations are low and time is critical.

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Differential gene expression in Escherichia coli during aerosolization from liquid suspension

Chao

et al. 2018

Appl Microbiol Biotechnol

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Comparative transcriptome analysis was used to determine the differentially expressed genes in Escherichia coli during aerosolization from liquid suspension. Isogenic mutant studies were then used to examine the potential part played by some of these genes in bacterial survival in the air. Bioaerosols were sampled after 3 min of nebulization, which aerosolized the bacteria from the liquid suspension to an aerosol chamber (A0), and after further 30 min of airborne suspension in the chamber (A30). Bacteria at A0 showed 65 differentially expressed genes (30 downregulated and 35 upregulated) as compared to the original bacteria in the nebulizer. Droplet evaporation models predicted a drop in temperature in the bioaerosols, which coincides with the change in the expression of cold shock protein genes-cspB and cspG in the bacteria. The most notable group of differentially expressed genes was sorbitol transport and metabolism genes (srlABDEMR). Other genes associated with osmotic stress, nutrient limitation, DNA damage, and other stresses were differentially expressed in the bacteria at A0. After further airborne suspension, one gene (ypfM, which encodes a hypothetical protein with unknown function) was downregulated in the bacteria at A30 as compared to those at A0. Finally, isogenic mutants with either the dps or srlA gene deleted (both genes were upregulated at A0) had lower survival than the parental strain, which is a sign of their potential ability to protect the bacteria in the air.

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Release of Free DNA by Membrane-Impaired Bacterial Aerosols Due to Aerosolization and Air Sampling

Cited by 79 publications

References 60 publications

Application of ATP-based bioluminescence for bioaerosol quantification: Effect of sampling method

Application of ATP-based bioluminescence for bioaerosol quantification: Effect of sampling method

Design and Evaluation of the Field-Deployable Electrostatic Precipitator with Superhydrophobic Surface (FDEPSS) with High Concentration Rate

Differential gene expression in Escherichia coli during aerosolization from liquid suspension

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