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Identify putative volatile biomarkers of Valley fever using a murine lung infection model (Human studies)

ABSTRACT: Coccidioides immitis and Coccidioides posadasii are soil-dwelling fungi of arid regions in North and South America that are responsible for Valley fever (coccidioidomycosis). Forty percent of patients with Valley fever exhibit symptoms ranging from mild, self-limiting respiratory infections, to severe, life-threatening pneumonia that requires treatment. Misdiagnosis as bacterial pneumonia commonly occurs in symptomatic Valley fever cases, resulting in inappropriate treatment with antibiotics, increased medical costs, and delay in diagnosis. In this study, we explored the feasibility of developing breath-based diagnostics for Valley fever using lung specimens from persons with community-acquired pneumonia (CAP). To investigate potential volatile biomarkers of Valley fever that arise from host-pathogen interactions, we collected bronchoalveolar lavage fluid (BALF) and sputum from patients treated at Mayo Clinic in Scottsdale, Arizona for untargeted volatile metabolomics via solid phase microextraction (SPME) and two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC-TOFMS). We identified 244 total volatile organic compounds (VOCs). Using Random Forest, we identified a subset of these VOCs that were also able to separate Coccidioides positive samples from bacteria positive samples. The data presented here show that Coccidioides and/or the host produce volatile metabolites that may yield biomarkers for a Valley fever breath test that can detect Coccidioidal infection.

ORGANISM(S): Human Homo Sapiens

TISSUE(S): Bronchoalveolar Lavage

DISEASE(S): Coccidioidomycosis

SUBMITTER: Heather Bean

PROVIDER: ST002449 | MetabolomicsWorkbench | Sat Dec 17 00:00:00 GMT 2022

REPOSITORIES: MetabolomicsWorkbench

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Project description:The oomycete P. oligandrum is a soil-inhabiting parasite and predator of both fungi and oomycetes and uses hydrolytic enzymes extensively to penetrate and hydrolyze its host or prey. Other mechanisms have been overlooked, and we investigated whether P. oligandrum-produced volatile organic compounds (VOCs) could also be contributing to antagonism. VOCs have diverse functions, including contributing to antagonism in ecological interactions and potential applications in biocontrol. The growth-inhibiting activity of P. oligandrum VOCs was tested on P. myriotylum – a host or prey of P. oligandrum – coupled with electron microscopy, biochemical assays and transcriptomic analysis. The total VOCs produced by P. oligandrum reduced P. myriotylum growth by 80% and zoospore levels by 60%. GC-MS identified twenty-three VOCs, and methyl heptenone, D-limonene, 2-undecanone and 1-octanal were potent inhibitors of P. myriotylum growth and led to increased production of reactive oxygen species at a concentration that did not inhibit P. oligandrum growth. Exposure to the total VOCs of P. oligandrum led to shrinkage of P. myriotylum hyphae, and lysis of the cellular membranes and organelles. Transcriptomic analysis of P. myriotylum exposed to the P. oligandrum VOCs at increasing levels of growth inhibition showed initially a strong upregulation of putative detoxification related genes that was not maintained later during exposure to the VOCs. The inhibition of P. myriotylum growth continued after the exposure to the VOCs was discontinued and led to reduced leaf lesion size during P. myriotylum infection of its plant host. The VOCs produced by P. oligandrum could be another factor alongside hydrolytic enzymes contributing to its ecological role as a microbial antagonist where the concentration of the VOCs may be inhibitory in particular ecological niches such as in soil. The VOCs analyzed here may also be contributing to the biocontrol of diseases using P. oligandrum commercial preparations.

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Identify putative volatile biomarkers of Valley fever using a murine lung infection model (Human studies)

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