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Investigating the relationship between breath aerosol size and exhaled breath condensate (EBC) metabolomic content.

ABSTRACT: Exhaled breath aerosols contain valuable metabolomic content due to gas exchange with blood at the alveolar capillary interface in the lung. Passive and selective filtering of these aerosols and droplets may reduce the amount of saliva contaminants and serve as an aid to enhance targeted metabolomic content when sampled in exhaled breath condensate (EBC). It is currently unknown if breath aerosol size distribution affects the types or abundances of metabolites sampled through EBC. This pilot study uses a previously described hand-held human breath sampler device with varying notch filter geometries to redirect the trajectory of breath aerosols based on size. Ten notch filter lengths were simulated with the device to calculate the effect of filter length on the breath aerosol size distribution and the proportion of aerosols which make their way through to an EBC collection tube. From three notch filter lengths, we investigate metabolite content of various aerosol fractions. We analyzed the non-volatile fraction of breath condensate with high performance liquid chromatography-mass spectrometry for broad metabolite coverage. We hypothesize that: (1) increasing the length of the notch filter in this device will prevent larger aerosols from reaching the collection tube thus altering the breath aerosol size distribution sampled in EBC; and (2) there is not a systematic large-scale difference in EBC metabolomic content that correlates with breath aerosol size. From simulation results, particles typically larger than 10 µm were filtered out. This indicates that a longer notch filter in this device prevents larger particles from reaching the collection tube thus altering the aerosol particle size distribution. Most compounds were commonly present in all three filter lengths tested, and we did not see strong statistical evidence of systematic metabolite differences between breath aerosol size distributions.

SUBMITTER: Schmidt AJ

PROVIDER: S-EPMC7880424 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The non-invasive, quick, and safe collection of exhaled breath condensate makes it a candidate as a diagnostic matrix in personalized health monitoring devices. The lack of standardization in collection methods and sample analysis is a persistent limitation preventing its practical use. The collection method and hardware design are recognized to significantly affect the metabolomic content of EBC samples, but this has not been systematically studied. Here, we completed a series of experiments to determine the sole effect of collection temperature on the metabolomic content of EBC. Temperature is a likely parameter that can be controlled to standardize among different devices. The study considered six temperature levels covering two physical phases of the sample; liquid and solid. The use of a single device in our study allowed keeping saliva filtering and collector surface effects as constant parameters and the temperature as a controlled variable; the physiological differences were minimized by averaging samples from a group of volunteers and a period of time. After EBC collection, we used an organic solvent rinse to collect the non-water-soluble compounds from the condenser surface. This additional matrix enhanced metabolites recovery, was less dependent on temperature changes, and may possibly serve as an additional pointer to standardize EBC sampling methodologies. The collected EBC samples were analyzed with a set of mass spectrometry methods to provide an overview of the compounds and their concentrations present at each temperature level. The total number of volatile and polar non-volatile compounds slightly increased in each physical phase as the collection temperature was lowered to minimum, 0?°C for liquid and -30, -56?°C for solid. The low-polarity non-volatile compounds showed a weak dependence on the collection temperature. The metabolomic content of EBC samples may not be solely dependent on temperature but may be influenced by other phenomena such as greater sample dilution due to condensation from the ambient air at colder temperatures, or due to adhesion properties of the collector surface and occurring chemical reactions. The relative importance of other design parameters such as condenser coating versus temperature requires further investigation.

Project description:The lung microbiota is commonly sampled using relatively invasive bronchoscopic procedures. Exhaled breath condensate (EBC) collection potentially offers a less invasive alternative for lung microbiota sampling. We compared lung microbiota samples retrieved by protected specimen brushings (PSB) and exhaled breath condensate collection. We also sought to assess whether aerosolized antibiotic treatment would influence the lung microbiota and whether this change could be detected in EBC. EBC was collected from 6 conscious sheep and then from the same anesthetized sheep during mechanical ventilation. Following the latter EBC collection, PSB samples were collected from separate sites within each sheep lung. On the subsequent day, each sheep was then treated with nebulized colistimethate sodium. Two days after nebulization, EBC and PSB samples were again collected. Bacterial DNA was quantified using 16S rRNA gene quantitative PCR. The V2-V3 region of the 16S rRNA gene was amplified by PCR and sequenced using Illumina MiSeq. Quality control and operational taxonomic unit (OTU) clustering were performed with mothur. The EBC samples contained significantly less bacterial DNA than the PSB samples. The EBC samples from anesthetized animals clustered separately by their bacterial community compositions in comparison to the PSB samples, and 37 bacterial OTUs were identified as differentially abundant between the two sample types. Despite only low concentrations of colistin being detected in bronchoalveolar lavage fluid, PSB samples were found to differ by their bacterial compositions before and after colistimethate sodium treatment. Our findings indicate that microbiota in EBC samples and PSB samples are not equivalent.IMPORTANCE Sampling of the lung microbiota usually necessitates performing bronchoscopic procedures that involve a hospital visit for human participants and the use of trained staff. The inconvenience and perceived discomfort of participating in this kind of research may deter healthy volunteers and may not be a safe option for patients with advanced lung disease. This study set out to evaluate a less invasive method for collecting lung microbiota samples by comparing samples taken via protected specimen brushings (PSB) to those taken via exhaled breath condensate (EBC) collection. We found that there was less bacterial DNA in EBC samples compared with that in PSB samples and that there were differences between the bacterial communities in the two sample types. We conclude that while EBC and PSB samples do not produce equivalent microbiota samples, the study of the EBC microbiota may still be of interest.

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Investigating the relationship between breath aerosol size and exhaled breath condensate (EBC) metabolomic content.

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