Summary of Study ST001659

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001064. The data can be accessed directly via it's Project DOI: 10.21228/M85H6W This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001659
Study TitleIdentify putative volatile biomarkers of Coccidioides spp. grown in vitro
Study TypeUntargeted metabolomics
Study SummaryValley fever (coccidioidomycosis) is an endemic fungal pneumonia of the North and South American deserts. The causative agents of Valley fever are the dimorphic fungi Coccidioides immitis and C. posadasii, which grow as mycelia in the environment and spherules within the lungs of vulnerable hosts. The current diagnostics for Valley fever are severely lacking due to poor sensitivity and invasiveness, contributing to a 23-day median time-to-diagnosis, and therefore new diagnostic tools are needed. We are working toward the development of a breath-based diagnostic for coccidioidomycosis, and in this initial study we characterized the volatile metabolomes (or volatilomes) of in vitro cultures of Coccidioides. Using solid-phase microextraction and comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC–TOFMS), we characterized the VOCs produced by six strains of each species during mycelial or spherule growth. We detected a total of 353 VOCs that were at least two-fold more abundant in a Coccidioides culture versus medium controls and found the volatile metabolome of Coccidioides is more dependent on growth phase (spherule versus mycelia) than on the species. The volatile profiles of C. immitis and C. posadasii have strong similarities, indicating that a single suite of Valley fever breath biomarkers can be developed to detect both species.
Institute
Arizona State University
DepartmentSchool of Life Sciences
LaboratoryBean Laboratory
Last NameBean
First NameHeather
AddressPO Box 874501 Tempe, AZ 85287
EmailHeather.D.Bean@asu.edu
Phone4807273395
Submit Date2021-01-22
PublicationsLifecycle dominates the volatilome character of the dimorphic fungus Coccidioides spp Emily A. Higgins Keppler, Heather L. Mead, Bridget M. Barker, Heather D. Bean bioRxiv 2021.01.15.426916; doi: https://doi.org/10.1101/2021.01.15.426916
Raw Data AvailableYes
Raw Data File Type(s)smp
Analysis Type DetailGC-MS
Release Date2021-03-15
Release Version1
Heather Bean Heather Bean
https://dx.doi.org/10.21228/M85H6W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Collection:

Collection ID:CO001729
Collection Summary:All Coccidioides isolates were grown under BSL-3 containment, using conditions that induce mycelial or spherule growth. For mycelial growth, a 50 ml vented falcon tube containing 10 ml of RPMI media (filter sterilized RPMI 1640, 10% fetal bovine serum) was inoculated with a 1 cm x 1 cm 2xGYE agar plug for each strain. These plates were inoculated using 100 µl of glycerol stock, spread across the plate, and cultured for 30°C for two weeks. Control RPMI media was inoculated with a plug from sterile 2xGYE agar media. Each sample, including media control, was prepared in triplicate. Cultures were grown on a shaking incubator at 150 rpm, 30°C for 96 h. For spherule cultures, a 50 ml vented falcon tube containing 10 ml of RPMI media was inoculated to a final concentration of 1.0 x 10^5 arthroconidia/ml in 1xPBS. Arthroconidia were grown and harvested. Strains RMSCC2343 and RMSCC3505 did not produce enough conidia to achieve 1.0 x 10^5 arthroconidia/ml, and were inoculated at 7.0 x 10^4 and 4.0 x 10^4 arthroconidia/ml, respectively. Control media was inoculated with 1 ml of sterile 1xPBS. Cultures were grown on a shaking incubator at 150 rpm, at 39°C in 10% CO2 for 96 h. Mycelial and spherule cultures were spun at 12,000 x g at 4°C for 10 min to pellet the cells. The supernatant was removed and place in a Nanosep MF Centrifugal Devices with Bio-Inert® Membrane 0.2 µm spin filter and centrifuged at 3,200 x g for 4 min. The filtrate was stored at −80°C until volatile metabolomics analysis. The Coccidioides spp. culture filtrates and media blanks were allowed to thaw at 4°C overnight, and then 2 ml were transferred and sealed into sterilized 10 ml GC headspace vials with PTFE/silicone septum screw caps. All samples were stored for up to 12 d at 4°C until analyzed.
Sample Type:Fungi cells
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