Breath metabolome of mice infected with Pseudomonas aeruginosa
The measurement of specific volatile organic compounds in breath has been proposed as a potential diagnostic for a variety of diseases. The most well-studied bacterial lung infection in the breath field is that caused by Pseudomonas aeruginosa.
To determine a discriminatory core of molecules in the “breath-print” of mice during a lung infection with four strains of P. aeruginosa (PAO1, PA14, PAK, PA7). Furthermore, we attempted to extrapolate a strain-specific “breath-print” signature to investigate the possibility of recapitulating the genetic phylogenetic groups (Stewart et al. Pathog Dis 71(1), 20–25, 2014. https://doi.org/10.1111/2049-632X.12107).
Breath was collected into a Tedlar bag and shortly after drawn into a thermal desorption tube. The latter was then analyzed into a comprehensive multidimensional gas chromatography coupled with a time-of-flight mass spectrometer. Random forest algorithm was used for selecting the most discriminatory features and creating a prediction model.
Three hundred and one molecules were significantly different between animals infected with P. aeruginosa, and those given a sham infection (PBS) or inoculated with UV-killed P. aeruginosa. Of those, nine metabolites could be used to discriminate between the three groups with an accuracy of 81%. Hierarchical clustering showed that the signature from breath was due to a specific response to live bacteria instead of a generic infection response. Furthermore, we identified ten additional volatile metabolites that could differentiate mice infected with different strains of P. aeruginosa. A phylogram generated from the ten metabolites showed that PAO1 and PA7 were the most distinct group, while PAK and PA14 were interspersed between the former two groups.
To the best of our knowledge, this is the first study to report on a ‘core’ murine breath print, as well as, strain level differences between the compounds in breath. We provide identifications (by running commercially available analytical standards) to five breath compounds that are predictive of P. aeruginosa infection.
KeywordsBreath Volatile organic compounds (VOCs) Pseudomonas aeruginosa Comprehensive gas chromatography-time-of-flight mass spectrometer (GC×GC ToF MS)
Financial support for this work was provided by Hitchcock Foundation and the National Institutes of Health (NIH, Project # R21AI12107601). MN and CAR were supported by the Burroughs Wellcome Fund institutional program grant unifying population and laboratory based sciences to Dartmouth College (Grant#1014106). CAR was additionally supported by a T32 training grant (T32LM012204, PI: Tor D Tosteson).
Conception and design: GP, JEH, LKAL, MJW; Acquisition of the data: GP, MN, FAF, CR, AM, DN; Analysis and interpretation of the data: MN, GP, FAF; Writing and review of the manuscript: GP, MN, FAF, CR, MJW, LKAL, JEH.
Compliance with ethical standards
Conflict of interest
All authors report no potential conflicts of interest.
This article does not contain any studies with human participants performed by any of the authors.
Research involving animal rights
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
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