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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