Microbial Environment of Spacecraft
Duane L. Pierson, Ph.D.NASA Johnson Space Center
April 2007
Duane L. Pierson, Ph.D.NASA Johnson Space Center
April 2007
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Microbiology LabJohnson Space Center-NASA
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• Infectious Diseases
• Allergies• Plant Diseases
• Material Degradation
• Immune Alteration
• Volatile Release• Food Spoilage
• Environmental Contamination
• Toxin Production
ADVERSE EFFECTS OF MICROORGANISMS
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MICROBIOLOGICAL RISKS
• Crewmembers Preflight screening, quarantine, vaccination, antimicrobials, antiviralsPreflight/inflight monitoring, biocidesPreflight analysesPreflight/inflight monitoring, filtrationPreflight/inflight monitoring, disinfectionPreflight cleaning, biosafety assessment, disinfection
• Water• Food• Air• Surfaces• Payloads
Sources Controls
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PATHOGENS
Staphylococcus aureusStreptococci sppEscherichia coliPseudomonas aeruginosaLegionella pneumophilaSalmonella sppHerpes virusesAspergillus sppCandida sppPenicillium spp
Mycobacterium tuberculosisCorynebacterium diphtheriaeBordetella pertussisNeisseria meningiditisSTDsHuman immunodeficiency virusHepatitis AHepatitis B and CInfluenza virusesExotic Viruses
Public Health Space Flight
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HEALTHHEALTH
DISEASEDISEASE
•• Crowded Living Conditions•• Closed-Loop Environment (Water/Air)•• Reduced Capability for Personal Hygiene•• Limited Clean-up and Disinfection Capability•• Inability to Isolate Contagious Crewmember•• Limited Treatment Capability and Crew Return•• Altered Immune Response
FACTORS INCREASING DISEASE RISK
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•• Crew Physical Examinations•• Immunization•• Health Stabilization Program•• Quarantine•• Preflight Food Testing•• Payload Biosafety Evaluation•• Establishment of Acceptability Limits•• Systems Design•• Environmental Monitoring•• In-Flight Housekeeping•• In-Flight Diagnostic Capabilities•• Antimicrobials
PREVENTIVE MEASURES
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10Isolation Frequency (%)
0 20 40 60 80 100
Staphylococcus sp.Micrococcus sp.
Bacillus sp.Corynebacterium sp.
Staphylococcus aureusStreptococcus sp.Acinetobacter sp.Enterobacter sp.
Unidentified gram negative rodsSerratia marcescens
Pseudomonas paucimobilisKlebsiella sp.
Flavobacterium sp.
Bacteria Isolated From Shuttle In-Flight Air (n=16)
11Isolation Frequency (%)
0 20 40 60 80 100
Penicillium sp.Aspergillus flavus
Aspergillus sp.Aspergillus nigerCladosporium sp.Unidentified yeastPaecilomyces sp.
Candida sp.Yarrowia sp.
HyphomyceteAcremonium sp.
Fungi Isolated From Air In The Dining Area (n=24)
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NASA-MIR
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14Location
0
200
400
600
800
1000
1200
SRV-Hot SRV-Cold SVO-ZV
Dominant Bacteria:Corynebacterium sp.Xanthomonas sp.CDC group EF-4Flavobacterium sp.
Col
ony
Form
ing
Uni
ts/1
00 m
LMir Water Data
Microbial Total Counts 1180
211
6.5
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Mir Water Isolates - SRV-K “Warm”Agrobacterium rhizogenesCDC Group EF-4CDC Group II-HClavibacter michiganenseCorynebacterium aquaticumFlavobacterium speciesFlavobacteriummeningosepticumFlavobacterium indologenesKingella kingaeKingella speciesKluvera ascorbataMethylobacterium extorquensPseudomonas aeruginosa
Psychrobacter glatheiRalstonia pickettiiRhizobium lotiSphingobacterium thalpophiliumSphingomonas paucimobilisSuttonella indologenesXanthomonas campestrisXanthomonas maltophiliaXanthomonas species
Microbial Concentration and Characteristics
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Gareth
Gareth Cook, Boston Globe Staff (10-1-00)
“… The aging Mir … is nearly overrun with the stuff (fungi). Visitors have found numerous fungal
patches with hues between green and black, feeding behind control panels, slowly digesting the ship’s air conditioner, communications unit, and myriad
other surfaces.”
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Wolf spent several hours working with Vinagradov to mop up a basketball-size drop of water…“I didn’t realize I bought myself anywhere from two to six hours per day doing this for the rest of the mission.”
From DRAGONFLY by Bryan Burrough
Microbial Concentration and CharacteristicsFloating Condensate – Case Study
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Acremonium speciesCandida guilliermondiiCandida kruseiCladosporium speciesFusarium speciesPenicillium speciesRhodotorula rubra
Candida guilliermondiiCandida lipolyticaFusarium speciesHansenula anomalaPenicillium speciesRhodotorula glutinisRhodotorula rubra
Candida guilliermondiiCandida lipolyticaCladosporium speciesFusarium speciesHansenula anomalaPenicillium speciesRhodotorula glutinisRhodotorula rubra
NASA 6 “SLIMY” SAMPLE
NASA 7 “SLIMY” SAMPLE
NASA 7 “FRESH” SAMPLE
MIR Condensate FungiMicrobial Concentration and Characteristics
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BIODEGRADATION
“…“…(fungi) feeding behind control (fungi) feeding behind control panels, slowly digesting the shippanels, slowly digesting the ship’’s s air conditioner, communications air conditioner, communications unit, and myriad other surfaces.unit, and myriad other surfaces.””Gareth Cook, Boston Globe Staff Gareth Cook, Boston Globe Staff (10(10--11--00)00)
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Davey and O’toole, MMBR, 2000
Dunne, Clin Microbiol Rev, 2002
21www.spaceflight.nasa.gov
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Food MicrobiologyAcceptability Limits
<20,000 CFU/gram<1 CFU/gram<1 CFU/gram <1 CFU/gram<1000 CFU/gram
Microbial Concentration and Characteristics
• Total aerobic count• Escherichia coli• Coagulase positive Staphylococci• Salmonella• Yeast and Molds
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Disqualified Food SamplesDisqualifying agentSample description
Enterobacter cloacae, Enterobacterintermedius, Pantoea agglomerans
Chicken saladAspergillus niger, Aspergillus fumigatusTrail mixYeast speciesJapanese sugar candyYeast speciesAlmond M&MsTotal aerobic (TNTC) Bacillus speciesOnion medley seasoningPseudomonas aeruginosa*Chicken saladTotal aerobic (TNTC) Bacillus speciesSan Francisco seasoningKlebsiella pneumoniae, Enterobacter cloacaeFreeze dried cornAspergillus fumigatus, Penicillium speciesFreeze dried chopped pecansEnterobacter cloacaeChicken Pineapple saladTotal aerobic (TNTC) Bacillus speciesBerry medleyStaphylococcus aureusMiso soupAspergillus flavusOatmeal with raisinsSalmonella species serotype "B"Freeze dried shrimp
Microbial Concentration and Characteristics
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Microbial Monitoring – Environment
• Air assessments• Surface assessments
• Multiple locations within the vehicles
• Hardware assessments• Random samples to
ensure quality control
• Payload assessments• Biosafety Review Board
Microbial Concentration and Characteristics
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IN-FLIGHT MICROBIAL SPECIFICATIONS AND MONITORING REQUIREMENTS FOR AIR
AND SURFACES
Acceptability LimitsSurfaces: Bacteria 10,000 CFU/100 cm2
Fungi 100 CFU/100 cm2
Air: Bacteria 1000 CFU/m3
Fungi 100 CFU/m3
Sampling FrequencyOnce per 3 months
Sampling LocationsSurfaces: 6 locationsAir: 3 locations
ModulesFGB Node 1 Service Module
U.S. Lab Node 2 Research Module 1Node 3 Research Module 2 U.S. Hab
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ISS Bacterial Isolates - Air
IBMP; Castro, et al, 2004
4/0410/0311/025/022/0210/019/016/015/01OrganismStaphylococcus auricularisStaphylococcus epidermidisAcinetobacter calcoaceticusCorynebacterium speciesStaphylococcus cohniiStaphylococcus hominisStaphylococcus saprophyticusStaphylococcus simulansStaphylococcus xylosisBacillus licheniformisStaphylococcus aureusStaphylococcus capitisStaphylococcus haemolyticusStreptococcus speciesCorynebacteriumafermemtansCorynebacterium riegeliiStaphylococcus speciesMicrococcus speciesStaphylococcus pumilisAcinetobacter lwoffii
5/03
Microbial Concentration and Characteristics
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ISS Bacterial Isolates - Air4/0410/0311/025/022/0210/019/016/015/01Organism
Staphylococcus auricularisStaphylococcus epidermidisAcinetobacter calcoaceticusCorynebacterium speciesStaphylococcus cohniiStaphylococcus hominisStaphylococcus saprophyticusStaphylococcus simulansStaphylococcus xylosisBacillus licheniformisStaphylococcus aureusStaphylococcus capitisStaphylococcus haemolyticusStreptococcus speciesCorynebacteriumafermemtansCorynebacterium riegeliiStaphylococcus speciesMicrococcus speciesStaphylococcus pumilisAcinetobacter lwoffii
5/03
IBMP; Castro, et al, 2004
Microbial Concentration and Characteristics
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ISS Bacterial Isolates - Air
4/0410/0311/025/022/0210/019/016/015/01OrganismStaphylococcus auricularisStaphylococcus epidermidisAcinetobacter calcoaceticusCorynebacterium speciesStaphylococcus cohniiStaphylococcus hominisStaphylococcus saprophyticusStaphylococcus simulansStaphylococcus xylosisBacillus licheniformisStaphylococcus aureusStaphylococcus capitisStaphylococcus haemolyticusStreptococcus speciesCorynebacteriumafermemtansCorynebacterium riegeliiStaphylococcus speciesMicrococcus speciesStaphylococcus pumilisAcinetobacter lwoffii
5/03
IBMP; Castro, et al, 2004
Microbial Concentration and Characteristics
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SURFACE ANALYSIS
• 40 sampling sessions performed
• Bacterial samples = 353 15 samples have exceeded acceptability
limit (10,000 colony forming units/100 cm2)Most common isolates – Staphylococcus and
Bacillus spp.
• Fungal samples = 31133 samples have exceeded acceptability
limit (1000 colony forming units/100 cm2)Most common isolates – Aspergillus and
Penicillium spp.
• Disinfection using Fungistat wipes performed
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Staphylococcus Sources - CrewMicrobial Concentration and Characteristics
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DNA Fingerprinting - S. aureus
• Blue - isolated from the crew of ISS-5, the crew of ISS-4, and in-flight environmental isolates
• Green - isolated from the crews of ISS-1, ISS-4, and ISS-5
• Orange - isolated from crewmembers of ISS-1 and ISS-4 and from an in-flight environmental surface
Microbial Concentration and Characteristics
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Biodegradation
• Microorganisms play a key role in the biodestruction of materials and can contribute to equipment and hardware malfunctions
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IN-FLIGHT MICROBIAL SPECIFICATIONS AND MONITORING REQUIREMENTS FOR
WATER
Acceptability Limits
Total Bacteria: 50 CFU/ml Coliforms: Non-detectable/100 ml
Sampling Locations Sampling FrequencyRussian Segment: Once every 3 months
SRV-K/Hot SRV-K/AmbientSVO-ZV
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Water Microbiology KitMicrobial Concentration and Characteristics
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Acidovorax temperansCorynebacterium speciesDechlorosoma suillumFlexibacter speciesMethylobacterium speciesMicrobacterium speciesPseudomonas aeruginosaRalstonia eutrophaRalstonia metalliduransRalstonia pickettiiSphingomonas paucimobilisSphingomonas stygialisSphingomonas xenophagaSphingomonas yanoikuyaeunidentified Gram negative rods
Leifsonia xyliMethylobacterium fugisawaenseMethylobacterium podariumMethylobacterium speciesPseudomonas fluorescensPseudomonas speciesRalstonia pickettiiSphingomonas speciesSphingomonas stygialisSphingomonas yanoikuyaeStenotrophomonas maltophiliaunidentified Gram negative rods
Organisms Isolated from ISS Water
SRV K Hot SRV K Warm SVO ZVAcidovorax temperansAcinetobacter radioresistensCaulobacter vibrioidesMethylobacterium speciesNon-viable organismsRalstonia eutrophaSphingomonas paucimobilisSphingomonas stygialisStaphylococcus species
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Microbiology Water Analysis Kit (MWAK)
•• Capability was initiated in MayCapability was initiated in May2004 to perform analysis for2004 to perform analysis forinin--flight detection of flight detection of coliformscoliformsin ISS potable and storedin ISS potable and storedwaterwater
•• Dry media is contained in Coliform Detection BagDry media is contained in Coliform Detection Bag
•• Direct bag to bag transfer of waterDirect bag to bag transfer of water
•• 44 +/44 +/-- 4 hours incubation period4 hours incubation period
•• Presence/Absence TestPresence/Absence Test
_ +
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Crew Exploration Vehicle
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Engineering/Design Solutions
• HEPA air filters• Moisture Control• Antimicrobial materials• In-line water filters• Surface disinfectants• Water biocides• Trash containment• Solid waste containment
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PREFLIGHT CREWMONITORING
PREFLIGHT CREWMONITORING
IN-FLIGHTMONITORING
IN-FLIGHTMONITORING
HOUSEKEEPINGHOUSEKEEPING
ACCEPTIBILITYLIMITS
ACCEPTIBILITYLIMITS
PREFLIGHTMONITORING
of WATER
PREFLIGHTMONITORING
of WATER
PREFLIGHTMONITORING
of FOODS
PREFLIGHTMONITORING
of FOODS
LIFESUPPORTSYSTEMS
LIFESUPPORTSYSTEMS
PREFLIGHTMONITORINGof PAYLOADS
PREFLIGHTMONITORINGof PAYLOADS
PREVENTION IS BETTER THAN REMEDIATIONPREVENTION IS BETTER THAN REMEDIATION
LESSONS LEARNED
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