#METABOLOMICS WORKBENCH ehiggins_20210122_122728 DATATRACK_ID:2415 STUDY_ID:ST001659 ANALYSIS_ID:AN002710 PROJECT_ID:PR001064 VERSION 1 CREATED_ON January 22, 2021, 1:24 pm #PROJECT PR:PROJECT_TITLE Volatile Biomarkers for a Valley Fever Breath Test PR:PROJECT_TYPE GCxGC-TOFMS metabolomics PR:PROJECT_SUMMARY Coccidioidomycosis, or valley fever, is prevalent in AZ, with more than 12,000 PR:PROJECT_SUMMARY new human infections diagnosed every year. In highly endemic areas, e.g., PR:PROJECT_SUMMARY Phoenix and Tucson, up to 30% of community-acquired pneumonia may be caused by PR:PROJECT_SUMMARY Valley fever, and cases are on the rise. The current diagnostics for Valley PR:PROJECT_SUMMARY fever are severely lacking due to invasiveness (biopsy) and poor sensitivity PR:PROJECT_SUMMARY (serology), strongly contributing to an unacceptable 23-day median PR:PROJECT_SUMMARY time-to-diagnosis. There is a critical need for sensitive and non-invasive PR:PROJECT_SUMMARY diagnostics for identifying Valley fever lung infections. Our long-term goal is PR:PROJECT_SUMMARY to substantially shorten the time-to-diagnosis for Valley fever through the PR:PROJECT_SUMMARY development of sensitive and specific breath-based diagnostics for PR:PROJECT_SUMMARY coccidioidomycosis lung infections. The overall objective of this application is PR:PROJECT_SUMMARY to identify and validate putative volatile biomarkers of Coccidioides infections PR:PROJECT_SUMMARY via metabolomics analyses of in vitro cultures, mouse model lung infections, and PR:PROJECT_SUMMARY lung specimens from humans with Valley fever. At the completion of the proposed PR:PROJECT_SUMMARY study, we expect to have identified and validated a panel of 10-15 volatile PR:PROJECT_SUMMARY biomarkers for the sensitive and specific detection of valley fever in lung PR:PROJECT_SUMMARY specimens. PR:INSTITUTE Arizona State University PR:DEPARTMENT School of Life Sciences PR:LABORATORY Bean Laboratory PR:LAST_NAME Bean PR:FIRST_NAME Heather PR:ADDRESS PO Box 874501, Tempe, AZ, 85287, USA PR:EMAIL Heather.D.Bean@asu.edu PR:PHONE 480-727-3395 PR:FUNDING_SOURCE Arizona Biomedical Research Centre New Investigator Award to HDB #STUDY ST:STUDY_TITLE Identify putative volatile biomarkers of Coccidioides spp. grown in vitro ST:STUDY_TYPE Untargeted metabolomics ST:STUDY_SUMMARY Valley fever (coccidioidomycosis) is an endemic fungal pneumonia of the North ST:STUDY_SUMMARY and South American deserts. The causative agents of Valley fever are the ST:STUDY_SUMMARY dimorphic fungi Coccidioides immitis and C. posadasii, which grow as mycelia in ST:STUDY_SUMMARY the environment and spherules within the lungs of vulnerable hosts. The current ST:STUDY_SUMMARY diagnostics for Valley fever are severely lacking due to poor sensitivity and ST:STUDY_SUMMARY invasiveness, contributing to a 23-day median time-to-diagnosis, and therefore ST:STUDY_SUMMARY new diagnostic tools are needed. We are working toward the development of a ST:STUDY_SUMMARY breath-based diagnostic for coccidioidomycosis, and in this initial study we ST:STUDY_SUMMARY characterized the volatile metabolomes (or volatilomes) of in vitro cultures of ST:STUDY_SUMMARY Coccidioides. Using solid-phase microextraction and comprehensive ST:STUDY_SUMMARY two-dimensional gas chromatography coupled to time-of-flight mass spectrometry ST:STUDY_SUMMARY (GC×GC–TOFMS), we characterized the VOCs produced by six strains of each ST:STUDY_SUMMARY species during mycelial or spherule growth. We detected a total of 353 VOCs that ST:STUDY_SUMMARY were at least two-fold more abundant in a Coccidioides culture versus medium ST:STUDY_SUMMARY controls and found the volatile metabolome of Coccidioides is more dependent on ST:STUDY_SUMMARY growth phase (spherule versus mycelia) than on the species. The volatile ST:STUDY_SUMMARY profiles of C. immitis and C. posadasii have strong similarities, indicating ST:STUDY_SUMMARY that a single suite of Valley fever breath biomarkers can be developed to detect ST:STUDY_SUMMARY both species. ST:INSTITUTE Arizona State University ST:DEPARTMENT School of Life Sciences ST:LABORATORY Bean Laboratory ST:LAST_NAME Bean ST:FIRST_NAME Heather ST:ADDRESS PO Box 874501 Tempe, AZ 85287 ST:EMAIL Heather.D.Bean@asu.edu ST:PHONE 4807273395 ST:PUBLICATIONS Lifecycle dominates the volatilome character of the dimorphic fungus ST:PUBLICATIONS Coccidioides spp Emily A. Higgins Keppler, Heather L. Mead, Bridget M. Barker, ST:PUBLICATIONS Heather D. Bean bioRxiv 2021.01.15.426916; doi: ST:PUBLICATIONS https://doi.org/10.1101/2021.01.15.426916 #SUBJECT SU:SUBJECT_TYPE Fungi SU:SUBJECT_SPECIES Coccidioides posadasii;Coccidioides immitis #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS C. posadasii B3221 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=B3221 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3221 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=B3221 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3221 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=B3221 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3221 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=B3221 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3221 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=B3221 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3221 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=B3221 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3222 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=B3222 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3222 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=B3222 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3222 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=B3222 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3222 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=B3222 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3222 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=B3222 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii B3222 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=B3222 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS Media Control Mycelia_2 treatment:Blank | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=Control Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS Media Control Mycelia_3 treatment:Blank | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=Control Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS Media Control Mycelia_4 treatment:Blank | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=Control Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS Media Control Spherule_1 treatment:Blank | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=Control Spherule_1.SMP SUBJECT_SAMPLE_FACTORS Media Control Spherule_2 treatment:Blank | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=Control Spherule_2.SMP SUBJECT_SAMPLE_FACTORS Media Control Spherule_3 treatment:Blank | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=Control Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii GT-166 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=GT-166 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii GT-166 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=GT-166 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii GT-166 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=GT-166 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. posadasii GT-166 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=GT-166 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii GT-166 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=GT-166 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii GT-166 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=GT-166 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2006 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2006 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2006 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2006 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2006 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2006 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2006 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2006 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2006 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2006 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2006 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2006 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2009 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2009 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2009 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2009 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2009 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2009 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2009 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2009 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2009 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2009 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2009 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2009 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2010 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2010 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2010 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2010 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2010 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2010 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2010 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2010 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2010 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2010 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2010 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2010 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC2343 Mycelia_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2343 Mycelia_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC2343 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2343 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC2343 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2343 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC2343 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2343 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC2343 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2343 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC2343 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2343 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2395 Mycelia_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2395 Mycelia_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2395 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2395 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2395 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC2395 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2395 Spherule _1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2395 Spherule _1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2395 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2395 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC2395 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC2395 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC3505 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC3505 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC3505 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC3505 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC3505 Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC3505 Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC3505 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC3505 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC3505 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC3505 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RMSCC3505 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC3505 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC3506 Mycelia_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC3506 Mycelia_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC3506 Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC3506 Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC3506 Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RMSCC3506 Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC3506 Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC3506 Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC3506 Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC3506 Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii RMSCC3506 Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RMSCC3506 Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RS Mycelia_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RS Mycelia_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RS Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RS Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. immitis RS Mycelia_4 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=RS Mycelia_4.SMP SUBJECT_SAMPLE_FACTORS C. immitis RS Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RS Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. immitis RS Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RS Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. immitis RS Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=RS Spherule_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii Silveira Mycelia_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=Silveira Mycelia_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii Silveira Mycelia_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=Silveira Mycelia_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii Silveira Mycelia_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:30°C | culture_conditions:normoxia RAW_FILE_NAME=Silveira Mycelia_3.SMP SUBJECT_SAMPLE_FACTORS C. posadasii Silveira Spherule_1 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=Silveira Spherule_1.SMP SUBJECT_SAMPLE_FACTORS C. posadasii Silveira Spherule_2 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=Silveira Spherule_2.SMP SUBJECT_SAMPLE_FACTORS C. posadasii Silveira Spherule_3 treatment:Fungi | culture_time (h):96 | culture_medium:RPMI | culture_tempurature:39°C | culture_conditions:10% CO2 RAW_FILE_NAME=Silveira Spherule_3.SMP #COLLECTION CO:COLLECTION_SUMMARY All Coccidioides isolates were grown under BSL-3 containment, using conditions CO:COLLECTION_SUMMARY that induce mycelial or spherule growth. For mycelial growth, a 50 ml vented CO:COLLECTION_SUMMARY falcon tube containing 10 ml of RPMI media (filter sterilized RPMI 1640, 10% CO:COLLECTION_SUMMARY fetal bovine serum) was inoculated with a 1 cm x 1 cm 2xGYE agar plug for each CO:COLLECTION_SUMMARY strain. These plates were inoculated using 100 µl of glycerol stock, spread CO:COLLECTION_SUMMARY across the plate, and cultured for 30°C for two weeks. Control RPMI media was CO:COLLECTION_SUMMARY inoculated with a plug from sterile 2xGYE agar media. Each sample, including CO:COLLECTION_SUMMARY media control, was prepared in triplicate. Cultures were grown on a shaking CO:COLLECTION_SUMMARY incubator at 150 rpm, 30°C for 96 h. For spherule cultures, a 50 ml vented CO:COLLECTION_SUMMARY falcon tube containing 10 ml of RPMI media was inoculated to a final CO:COLLECTION_SUMMARY concentration of 1.0 x 10^5 arthroconidia/ml in 1xPBS. Arthroconidia were grown CO:COLLECTION_SUMMARY and harvested. Strains RMSCC2343 and RMSCC3505 did not produce enough conidia to CO:COLLECTION_SUMMARY achieve 1.0 x 10^5 arthroconidia/ml, and were inoculated at 7.0 x 10^4 and 4.0 x CO:COLLECTION_SUMMARY 10^4 arthroconidia/ml, respectively. Control media was inoculated with 1 ml of CO:COLLECTION_SUMMARY sterile 1xPBS. Cultures were grown on a shaking incubator at 150 rpm, at 39°C CO:COLLECTION_SUMMARY in 10% CO2 for 96 h. Mycelial and spherule cultures were spun at 12,000 x g at CO:COLLECTION_SUMMARY 4°C for 10 min to pellet the cells. The supernatant was removed and place in a CO:COLLECTION_SUMMARY Nanosep MF Centrifugal Devices with Bio-Inert® Membrane 0.2 µm spin filter and CO:COLLECTION_SUMMARY centrifuged at 3,200 x g for 4 min. The filtrate was stored at −80°C until CO:COLLECTION_SUMMARY volatile metabolomics analysis. The Coccidioides spp. culture filtrates and CO:COLLECTION_SUMMARY media blanks were allowed to thaw at 4°C overnight, and then 2 ml were CO:COLLECTION_SUMMARY transferred and sealed into sterilized 10 ml GC headspace vials with CO:COLLECTION_SUMMARY PTFE/silicone septum screw caps. All samples were stored for up to 12 d at 4°C CO:COLLECTION_SUMMARY until analyzed. CO:SAMPLE_TYPE Fungi cells #TREATMENT TR:TREATMENT_SUMMARY All Coccidioides isolates were grown under BSL-3 containment, using conditions TR:TREATMENT_SUMMARY that induce mycelial or spherule growth. See Collection Protocol for details #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Filtrate samples were heated to 50 degrees Celsius with agitation, and volatiles SP:SAMPLEPREP_SUMMARY were sampled for 10 minutes using solid-phase microextraction (SPME) and SP:SAMPLEPREP_SUMMARY analyzed using comprehensive gas chromatography coupled to time-of-flight mass SP:SAMPLEPREP_SUMMARY spectrometry. SP:EXTRACTION_METHOD Solid-phase microextraction (SPME) #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Samples analyzed using comprehensive two-dimensional gas chromatography (GCxGC). CH:CHROMATOGRAPHY_SUMMARY See attached method file. CH:CHROMATOGRAPHY_TYPE GCxGC CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Multidimensional column configuration #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Leco Pegasus 4D GCxGC TOF MS:INSTRUMENT_TYPE GC x GC-TOF MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS see attached MS methods MS:MS_RESULTS_FILE ST001659_AN002710_Results.txt UNITS:Peak areas Has m/z:No Has RT:Yes RT units:Seconds #END