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

Abstract

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.

Aerobiology: Experimental Considerations, Observations, and Future Tools

“…Aerosol generation and sampling prior to microbiological analysis are conducted for a range of bioaerosol-related research activities (e.g., determination of aerosol decay rates and inhalational infectious dose, efficacy of decontamination strategies, and evaluation of bioaerosol sampling technologies). These dynamic processes can cause damage due to shear forces acting on the microbial cells ( 12 – 27 ). Table 1 outlines some major aerosol generators and samplers used in aerobiological studies and their operating mechanisms.…”

“…However, comparative studies show that refluxing nebulizers produce the greatest loss of physiological function as a function of time in bacteria ( 16 , 19 – 21 , 24 ). The loss of function has been linked to membrane damage ( 13 , 20 , 24 ), release of ions into the medium (e.g., PO 4 2− ) ( 28 ), cell fragmentation ( 15 , 23 ), reduction in ATP activity ( 27 ), and magnitude of associated electrical charge ( 29 ), as the bacteria remaining in the nebulizer repeatedly pass through the device nozzles. Similar effects are observed for viruses ( 25 ).…”

Optimization of a Portable Adenosine Triphosphate Bioluminescence Assay Coupled with a Receiver Operating Characteristic Model to Assess Bioaerosol Concentrations on Site

“…To determine the ATP level in bioaerosols, applying a liquid medium for collecting bioaerosols was preferred over using an agar impactor when the ATP bioluminescence assay was conducted [ 2 , 12 , 16 , 18 ]. The application of an impactor and subsequent swabbing of the agar surface to obtain ATP levels may underestimate the ATP levels of bioaerosols.…”

Improvements of ATP Assay as a Substitute for the CFU Method in Estimating Viable Cell Count for BCG/rBCG Vaccine Preparations

“…In contrast to the multiweek CFU assay, it can be completed within 1 day. The ATP assay is based on the reaction of firefly luciferase with ATP which results in a bioluminescent product [6][7][8][9]. Since ATP is a major metabolite of living cells and is rapidly lost in dead cells, measuring ATP content can provide a reliable estimate of the number of living cells in a culture.…”