The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms
Ensuring biological cleanliness while assembling and launching spacecraft is critical for robotic exploration of the solar system. To date, when preventing forward contamination of other celestial bodies, NASA Planetary Protection policies have focused on endospore-forming bacteria while fungi were...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.777133/full |
| _version_ | 1818478822046040064 |
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| author | Adriana Blachowicz Snehit Mhatre Nitin Kumar Singh Jason M. Wood Ceth W. Parker Cynthia Ly Daniel Butler Christopher E. Mason Christopher E. Mason Kasthuri Venkateswaran |
| author_facet | Adriana Blachowicz Snehit Mhatre Nitin Kumar Singh Jason M. Wood Ceth W. Parker Cynthia Ly Daniel Butler Christopher E. Mason Christopher E. Mason Kasthuri Venkateswaran |
| author_sort | Adriana Blachowicz |
| collection | DOAJ |
| description | Ensuring biological cleanliness while assembling and launching spacecraft is critical for robotic exploration of the solar system. To date, when preventing forward contamination of other celestial bodies, NASA Planetary Protection policies have focused on endospore-forming bacteria while fungi were neglected. In this study, for the first time the mycobiome of two spacecraft assembly facilities at Jet Propulsion Laboratory (JPL) and Kennedy Space Center (KSC) was assessed using both cultivation and sequencing techniques. To facilitate enumeration of viable fungal populations and downstream molecular analyses, collected samples were first treated with chloramphenicol for 24 h and then with propidium monoazide (PMA). Among cultivable fungi, 28 distinct species were observed, 16 at JPL and 16 at KSC facilities, while 13 isolates were potentially novel species. Only four isolated species Aureobasidium melanogenum, Penicillium fuscoglaucum, Penicillium decumbens, and Zalaria obscura were present in both cleanroom facilities, which suggests that mycobiomes differ significantly between distant locations. To better visualize the biogeography of all isolated strains the network analysis was undertaken and confirmed higher abundance of Malassezia globosa and Cyberlindnera jadinii. When amplicon sequencing was performed, JPL-SAF and KSC-PHSF showed differing mycobiomes. Metagenomic fungal reads were dominated by Ascomycota (91%) and Basidiomycota (7.15%). Similar to amplicon sequencing, the number of fungal reads changed following antibiotic treatment in both cleanrooms; however, the opposite trends were observed. Alas, treatment with the antibiotic did not allow for definitive ascribing changes observed in fungal populations between treated and untreated samples in both cleanrooms. Rather, these substantial differences in fungal abundance might be attributed to several factors, including the geographical location, climate and the in-house cleaning procedures used to maintain the cleanrooms. This study is a first step in characterizing cultivable and viable fungal populations in cleanrooms to assess fungal potential as biocontaminants during interplanetary explorations. The outcomes of this and future studies could be implemented in other cleanrooms that require to reduce microbial burden, like intensive care units, operating rooms, or cleanrooms in the semiconducting and pharmaceutical industries. |
| first_indexed | 2024-12-10T09:52:56Z |
| format | Article |
| id | doaj.art-30f0f2d20ea04b8489152314fc972299 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-10T09:52:56Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-30f0f2d20ea04b8489152314fc9722992022-12-22T01:53:36ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-04-011310.3389/fmicb.2022.777133777133The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly CleanroomsAdriana Blachowicz0Snehit Mhatre1Nitin Kumar Singh2Jason M. Wood3Ceth W. Parker4Cynthia Ly5Daniel Butler6Christopher E. Mason7Christopher E. Mason8Kasthuri Venkateswaran9Biotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesBiotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesBiotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesBiotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesBiotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesBiotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesDepartment of Physiology and Biophysics, Weill Cornell Medicine, New York, NY, United StatesDepartment of Physiology and Biophysics, Weill Cornell Medicine, New York, NY, United StatesThe WorldQuant Initiative for Quantitative Prediction, Weill Cornell Medicine, New York, NY, United StatesBiotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, United StatesEnsuring biological cleanliness while assembling and launching spacecraft is critical for robotic exploration of the solar system. To date, when preventing forward contamination of other celestial bodies, NASA Planetary Protection policies have focused on endospore-forming bacteria while fungi were neglected. In this study, for the first time the mycobiome of two spacecraft assembly facilities at Jet Propulsion Laboratory (JPL) and Kennedy Space Center (KSC) was assessed using both cultivation and sequencing techniques. To facilitate enumeration of viable fungal populations and downstream molecular analyses, collected samples were first treated with chloramphenicol for 24 h and then with propidium monoazide (PMA). Among cultivable fungi, 28 distinct species were observed, 16 at JPL and 16 at KSC facilities, while 13 isolates were potentially novel species. Only four isolated species Aureobasidium melanogenum, Penicillium fuscoglaucum, Penicillium decumbens, and Zalaria obscura were present in both cleanroom facilities, which suggests that mycobiomes differ significantly between distant locations. To better visualize the biogeography of all isolated strains the network analysis was undertaken and confirmed higher abundance of Malassezia globosa and Cyberlindnera jadinii. When amplicon sequencing was performed, JPL-SAF and KSC-PHSF showed differing mycobiomes. Metagenomic fungal reads were dominated by Ascomycota (91%) and Basidiomycota (7.15%). Similar to amplicon sequencing, the number of fungal reads changed following antibiotic treatment in both cleanrooms; however, the opposite trends were observed. Alas, treatment with the antibiotic did not allow for definitive ascribing changes observed in fungal populations between treated and untreated samples in both cleanrooms. Rather, these substantial differences in fungal abundance might be attributed to several factors, including the geographical location, climate and the in-house cleaning procedures used to maintain the cleanrooms. This study is a first step in characterizing cultivable and viable fungal populations in cleanrooms to assess fungal potential as biocontaminants during interplanetary explorations. The outcomes of this and future studies could be implemented in other cleanrooms that require to reduce microbial burden, like intensive care units, operating rooms, or cleanrooms in the semiconducting and pharmaceutical industries.https://www.frontiersin.org/articles/10.3389/fmicb.2022.777133/fullmycobiomecleanroomchloramphenicolmetagenomeamplicon sequencing |
| spellingShingle | Adriana Blachowicz Snehit Mhatre Nitin Kumar Singh Jason M. Wood Ceth W. Parker Cynthia Ly Daniel Butler Christopher E. Mason Christopher E. Mason Kasthuri Venkateswaran The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms Frontiers in Microbiology mycobiome cleanroom chloramphenicol metagenome amplicon sequencing |
| title | The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms |
| title_full | The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms |
| title_fullStr | The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms |
| title_full_unstemmed | The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms |
| title_short | The Isolation and Characterization of Rare Mycobiome Associated With Spacecraft Assembly Cleanrooms |
| title_sort | isolation and characterization of rare mycobiome associated with spacecraft assembly cleanrooms |
| topic | mycobiome cleanroom chloramphenicol metagenome amplicon sequencing |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.777133/full |
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