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Experimental parameters defining ultra-low biomass bioaerosol analysis.

ABSTRACT: Investigation of the microbial ecology of terrestrial, aquatic and atmospheric ecosystems requires specific sampling and analytical technologies, owing to vastly different biomass densities typically encountered. In particular, the ultra-low biomass nature of air presents an inherent analytical challenge that is confounded by temporal fluctuations in community structure. Our ultra-low biomass pipeline advances the field of bioaerosol research by significantly reducing sampling times from days/weeks/months to minutes/hours, while maintaining the ability to perform species-level identification through direct metagenomic sequencing. The study further addresses all experimental factors contributing to analysis outcome, such as amassment, storage and extraction, as well as factors that impact on nucleic acid analysis. Quantity and quality of nucleic acid extracts from each optimisation step are evaluated using fluorometry, qPCR and sequencing. Both metagenomics and marker gene amplification-based (16S and ITS) sequencing are assessed with regard to their taxonomic resolution and inter-comparability. The pipeline is robust across a wide range of climatic settings, ranging from arctic to desert to tropical environments. Ultimately, the pipeline can be adapted to environmental settings, such as dust and surfaces, which also require ultra-low biomass analytics.

SUBMITTER: Luhung I

PROVIDER: S-EPMC8052325 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundDuring medical procedures with the potential to produce aerosols such as bronchoscopy, intubation, or CPR, health-care workers (HCWs) may be exposed to infectious bioaerosols. This scenario is of particular concern when high consequence pathogens such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are circulating. Thousands of HCWs have been infected with SARS-CoV-2. However, the determinants of aerosol generation during medical procedures and their relative risk to HCWs remain poorly characterized.Research questionThe goal of this study was to characterize aerosols produced during airway intubation by using an uninfected translational animal model and in human subjects undergoing elective aerosol-generating procedures. The study also determined the particle size distribution of generated particles.Study design and methodsAerosol generation was measured during highly controlled experimental (pig) intubations (N = 16) and elective bronchoscopies in uninfected patients (N = 49) using an optical particle counter. Recovery of normal respiratory flora was used as a surrogate for pathogen dispersion.ResultsThere was a small but significant (P = .03) decrease in 0.3 μm size particles during highly controlled pig intubations compared with baseline. The concentration of 1.0 μm and 5.0 μm aerosol particles did not significantly change, although oral bacteria were collected from the air. For elective patient bronchoscopies, there was a significant decrease in the generation of larger particles (1.0 μm and 5.0 μm) compared with baseline (P < .01); however, 18 of 39 (46%) patients showed increased aerosol production in 0.3 μm size particles, four of whom exhibited measurable increases.InterpretationAlthough the total amount of aerosols produced during intubation and bronchoscopy did not increase significantly relative to preprocedural levels, a small number of participants exhibited a measurable increase in submicron particle emission, meriting further research to delineate determinants of fine particle production during aerosol-generating procedures.

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Experimental parameters defining ultra-low biomass bioaerosol analysis.

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