Comprehensive Microbial Diversity Characterization of the International Space Station and Analogous Closed Habitats on Earth
Jessica Cisneros1,2, Parag Vaishampayan1, Pablo de Leon3, Jay Perry4, Kasthuri Venkateswaran1
1 Biotechnology and Planetary Protection Group, Jet Propulsion Laboratory, California Institute of Technology, USA
2 Department of Biology, California State University, Los Angeles
3 University of North Dakota, Grand Forks, USA
4 Marshall Space Flight Center, Huntsville, AL* Contact: [email protected]
Acknowledgements
Part of the research described in this study was carried out at the Jet Propulsion Laboratory, California Institute of
Technology, under contract with the National Aeronautics and Space Administration. This research was funded by
a 2012 Space Biology NNH12ZTT001N grant # 19-12829-26 under Task Order NNN13D111T award to K.
Venkateswaran. The authors gratefully acknowledge ISS Expedition 31 crew. Cisneros, a Louis Stokes Alliance
for Minority Participation – Bridge to the Doctorate (LSAMP-BD) fellow, was supported by the LSAMP-BD (Cohort
X) program (National Science Foundation grant # HRD-1246662).
Abstract
• Maintaining safety of astronauts during long-term habitation in the International Space Station (ISS) is
an essential undertaking that must be attained for sustained human presence in space.
• Of great concern is the ubiquitous presence of microorganisms, which have the potential to not only
affect crew health but also have adverse effects on crew systems. Increased microbial burden on ISS
could lead to infection or disease outbreaks in the closed habitat.
• The threat of such an outbreak is especially hazardous due to the limited treatment and patient
quarantine options.
• To date, monitoring the microbial diversity of the ISS was performed utilizing culture-based methods
for detailed molecular characterization.
Observations and Conclusions
Methodology
Data obtained from the 454
pyrosequencing was subjected to
computational analysis to estimate the
microbial diversity and community
differences among the three
environments .
High-throughput pyrosequencing
(454)
Results
Viable cells
PMA/Light PCR
Non-viable cellsPMA intercalated DNA:
non amplifiable by PCR
Viable cell selection
DNA Extraction
Promega.com
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
1.00E+08
1.00E+09
ILMH-0 ILMH-10
16
S rR
NA
co
py
nu
mb
er/
1 m
2
D.
21%
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
1.00E+08
1.00E+09
ISS-Lint ISS-Debris SAF-Lint SAF-debris
16
S rR
NA
co
py
nu
mb
er /
g
C.
Figure A-B: Total and viable microbial burden as estimated by the ATP assay. The viable microbial population was higher in ISS when compared to SAF samples.Figure C-D: Total and viable bacterial burden as estimated by the qPCR assay. In contrast to ATP assay, the measurement estimated by PMA-qPCR exhibited higher viable bacterial population in SAF (~45-75%) when compared to ISS (3-11%) samples. This results show that ISS debris samples might have more fungal population than the SAF samples where the high pH cleaning agents used might be detrimental for fungal survival.
Sample Collection Sites
Habitat – Specific bacterial Phyla as depicted by heat map. ISS habitat samples were enriched with Actinobacteria, SAF samples contained
predominately Proteobacteria, and ILMH surface locations were abundant with Firmicute members. Since ILMH air is circulated with external air,
environmental microbes such as Firmicutes were observed. Likewise, high human traffic were documented in SAF thus presence of
Proteobacteria was expected. Unlike these two Earth analogues, ISS was a closed habitat and robust Actinobacterial group was evident.
• Percent abundance of OTUs given by their Phyla. Samples treated with and without PMA.
• Similar trends with Proteobacteria abundance was observed with SAF and ILMH locations
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
1.00E+08
ISS-Lint ISS-Debris SAF-Lint SAF-Debris
RLU
/g 34% 86% 9% 31%
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
1.00E+08
ILMH-0 ILMH-10 PBS
RLU
/cm
2
17% 72% 37%
• The microbial diversity of three distinct closed habitats such as the ISS, Inflatable Lunar-Mars Habitat
(ILMH) at University of North Dakota and a NASA spacecraft assembly facility (SAF) was characterized
using the state-of-the-art molecular techniques.
• Particles accumulated in the vacuum cleaner bags (ISS and SAF) and the floor samples of ILMH were
aseptically sampled and analyzed.
• Cultivation techniques augmented with rapid molecular techniques were used to measure total, viable,
and cultivable microbial burden.
• ATP assays was used to estimate the total and viable microbial burden.
• Propidium monoazide (PMA) treatment followed by qPCR was used to estimate viable bacterial burden.
• In addition, next-generation sequencing technologies were used to elucidate the full spectrum of total
and viable bacterial, archaeal, and fungal diversity.
3% 11% 75% 44%
Actinobacteria
Firmicutes
Proteobacteria
Bacterial Phyla Bacterial Agent
example
Propionibacterium acnes Acne disease
Staphylococcus epidermidis Skin rashes
Methylobacterium No disease
Disease
-2 0 2
Co
un
t
02
00
40
0
Total Viable
ISS
Total Viable
SAF
Total Viable
ILMH
Total Viable
ISS
Total Viable
SAF
Total Viable
ILMH
Total Viable
ISS
Total Viable
SAF
Total Viable
ILMH
• As estimated by ATP assay, the viable microbial burden of the debris collected from the ISS
are ~1 log higher than observed in SAF cleanroom environments. SAF floors are regularly
maintained with high pH cleaning agents where as such cleaning exercises are not routinely
performed in ISS unless otherwise any problem arise.
• Prior to human occupation, the ILMH surfaces were kept impeccably clean due to rigorous
maintenance. Subsequent 10-days ILMH occupation by “student astronauts” influenced
increase in viable microbial (106 RLU per m2; ATP-assay) and bacterial (108 rrn per m2; PMA-
qPCR-assay) burden in these ILMH surfaces. The source of these microbes might be
attributed to the environmental control system (air exchange) and human shedding.
• Two logs of higher bacterial burden as estimated by PMA-qPCR need further research. One
possible explanation for the higher bacterial signal is that by average ~10 rrn copies might be
present in a given bacterial cell.
• The approach adapted during this study was the first ever-comprehensive molecular effort to
assess the viability of microbial cells associated with space-related enclosed habitat and to
correlate differential viability with phylogenetic affiliation.
• The results of this study also improved our understanding of background microbial
contamination, thus facilitating the development of biosensors to monitor closed habitats like
ISS and future manned missions.
The ILMH surfaces exhibited accumulation of ~106 microbes and ~108 bacterial population per m2 after 10 days of “student astronauts”
occupation. Among these population ~70% microbial and ~21% bacterial populations are viable.
Legend:
Dataset HM
0
6
12
20
4913
9806
14700
Color ranges:
Proteobacteria
Other
Actinobacteria
Firmicutes
Bacteroidetes
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Legend:
Dataset HM
0
6
12
20
4913
9806
14700
Color ranges:
Proteobacteria
Other
Actinobacteria
Firmicutes
Bacteroidetes
Acidobacteria
IOD
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IJD
D03G
7P
VX
IOD
IJDD
03F
U6W
0
IOD
IJDD
03F
UX
DF
IOD
IJD
D03G
KC
GP
IEP
H9V
M01C
6T
0M
ILA
NS
YW
03G
XA
31
IOD
IJD
D03G
NQ
R0
ILA
NS
YW
03G
4F
JA
ILA
NS
YW
03H
E5M
I
IOD
IJD
D03H
FQ
8I
IEP
H9V
M01E
BC
Q1
IOD
IJD
D03G
KT
1W
IOD
IJD
D03H
AM
HE
IEP
H9V
M01B
BG
Z2
IOD
IJD
D03F
T3Y
9
IOD
IJD
D0
3H
DJT
2
IOD
IJD
D03
G5
CD
1
IOD
IJD
D0
3G
577O
IOD
IJD
D0
3G
I1Q
S
IOD
IJD
D03
G7
HO
Y
IOD
IJD
D0
3G
CM
WS
IOD
IJD
D03G
QP
DV
IOD
IJD
D03G
UT
PF
IOD
IJD
D0
3G
C8
4D
ILA
NS
YW
03FX
W0J
ILA
NS
YW
03G
P0D
V
IOD
IJD
D03G
46U
D
IOD
IJD
D03G
06G
3
IOD
IJD
D03FTLM
K
IOD
IJD
D03G
MTM
7
IOD
IJD
D03
HA
VC
D
IOD
IJD
D03
FZZPL
IOD
IJD
D03
G8Q
I7
IOD
IJD
D03G
570C
IOD
IJD
D03G
NR
07
IODIJ
DD03
GPK03
IODIJ
DD03
HEK9F
IODIJ
DD03
F72G
A
IODIJ
DD03
G4W
1U
IODIJ
DD
03G
AK3MIO
DIJ
DD03
FSTV
G
IODIJ
DD03
GX8E
X
IODIJ
DD03
GM
HRS
IODIJ
DD03F
UG
04
IODIJ
DD03
GRN94
IOD
IJD
D03
GLN
JG
IOD
IJD
D03
HC
OXZ
IOD
IJD
D03
HG
FW
K
IOD
IJD
D03
GC
M50
IOD
IJD
D0
3F
0T
5O
IOD
IJD
D03
GK
N0S
ILA
NS
YW
03
GR
QU
C
IODIJD
D03G
ZKVR
IEPH9V
M01A
P392
IODIJD
D03G
BLJ8
IOD
IJDD
03G
9W
0B
IOD
IJDD
03G
AC
US
IOD
IJDD
03FO
SS
M
IOD
IJDD
03G
G4LG
IOD
IJDD
03F4N
MT
IOD
IJDD
03FS
52R
IOD
IJDD
03H
AC
LG
IOD
IJDD
03G
NX
QT
IOD
IJDD
03G
OO
LY
IOD
IJDD
03GA2Z
9
IOD
IJDD
03GC
DN
T
IOD
IJDD
03GG
SD
5
IOD
IJDD
03G0S
VI
IOD
IJDD
03GO
4QL
IODIJD
D03G
OK3X
IODIJD
D03G
90CQ
IODIJD
D03G
YX2S
IOD
IJDD
03GO
9BS
IODIJ
DD03H
C0FH
IODIJ
DD03FLIDO
IODIJD
D03FMEKW
ILANSYW
03GM
4CZ
ILAN
SYW03G
20IE
IEPH
9VM
01B62Z
F
ILA
NS
YW
03H
HO
6L
IEP
H9V
M01
DM
SY
4
ILAN
SYW
03H
CF9Y
ILAN
SYW
03F3Q
MV
ILAN
SYW
03F74W
L
IEP
H9V
M01A
5TK
Y
ILA
NS
YW
03F
5K
AF
IEP
H9V
M01D
JPR
K
IEP
H9V
M01E
CJI3
IEP
H9V
M01E
JVV
6
IEP
H9V
M01D
NH
JQ
IEP
H9V
M01E
CY
7C
ILA
NS
YW
03G
CR
2C
IEP
H9V
M01D
B64Y
IEP
H9V
M01B
6I2
2IE
PH
9V
M01D
WP
RL
IEP
H9V
M01B
EJO
TIE
PH
9V
M01C
27P
UIE
PH
9V
M01B
SB
5N
ILA
NS
YW
03
GQ
H5
AIE
PH
9V
M0
1D
ZE
2W
IEP
H9
VM
01E
ZN
W6
IEP
H9V
M01C
802I
IEP
H9
VM
01B
33E
L
ILA
NS
YW
03G
MW
CP
ILA
NS
YW
03FQ
Q8T
ILA
NS
YW
03H
GIZ
M
IEP
H9V
M01B
TS
KT
IEP
H9V
M01B
UV
7U
IEP
H9V
M01B
Z1M
N
IEP
H9V
M01D
IHU
0
ILAN
SYW
03FW
L9P
ILAN
SYW
03HD
R0Z
IEPH9V
M01C
6W5S
IEPH9V
M01D
9DNK
IEPH9V
M01C
P9K
1
ILAN
SYW
03GW
E9S
ILANSYW
03GPYAH
ILANSYW03FZO
FE
ILANSYW03G
KJTA
ILANSYW
03HB55O
ILANSYW
03GDZIZ
ILANSYW
03HE047
ILANSYW
03GM
3FBILANSYW03G
JCGN
ILANSYW
03G7C
UA
ILANSYW
03HBA8Y
ILANSYW03GC367
IODIJDD03GSW28
IODIJDD03FUAVU
IODIJDD03G7C0M
IODIJDD03GCDRPIODIJDD03G8F2Q
IODIJDD03HDG6F
IODIJDD03GJJGV
IODIJDD03F7W1E
IODIJDD03F771I
IODIJDD03GXZV1
IODIJDD03F3RJ5
ILANSYW03F6Q1O
ILANSYW03GX9GS
ILANSYW03G2X8B
ILANSYW03FX3NTILANSYW03G7HKA
IEPH9VM01EOSLJILANSYW03GDT24
ILANSYW03GZHSJILANSYW03FW4HCILANSYW03GSMGX
IEPH9VM01CXRQJ
ILANSYW03F0MTUIODIJDD03GZRJD
IODIJDD03GBWGL
IODIJDD03HENGG
IODIJDD03GC2D1
IODIJDD03HCZXSIODIJDD03F3GXE
IODIJDD03F9QQZ
ILANSYW03G3YPX
IODIJDD03FKDPIIODIJDD03GLHAW
IODIJDD03G2XVY
IODIJDD03G88PTIODIJDD03GK581
IODIJDD03FNCLN
IODIJDD03F0SMX
IODIJDD03F0OW8
IODIJDD03G7MWN
IODIJDD03FT36L
IODIJDD03G0SUS
IODIJDD03GC6ZX
IODIJDD03GSL2O
IODIJDD03FTU44
ILANSYW03GPLIM
ILANSYW03F2C38
IODIJDD03GCOUI
IODIJDD03GEBX6IODIJDD03FKGWIILANSYW03GL4J0IODIJDD03GS13V
IODIJDD03GQFMV
IODIJDD03GJ3L5IODIJDD03GI6JK
IODIJDD03G2J77
IODIJDD03G0TDW
IEPH9VM01DWFIYIEPH9VM01AKOEAIEPH9VM01EGIEI
ILANSYW03HDQ2J
IODIJDD03G1EGR
IODIJDD03F3MH1
IODIJDD03GSADN
IODIJDD03GT43M
IODIJDD03FUYTF
ILANSYW03GBUWZ
IODIJD
D03HDW9P
ILANSYW
03HD0LE
ILANSYW03GKVW
DIO
DIJDD03FZEHF
IODIJDD03GLI5X
IODIJDD03GE65U
IODIJDD03GRPMO
ILANSYW03F1SV7
IODIJDD03GWFKG
IODIJDD03GUESG
IODIJDD03G11ZUIODIJDD03FVHTW
IODIJDD03GK0NP
IODIJDD03GIJGL
IODIJDD03HB70ZIODIJDD03GLJ9Z
IEPH9VM01BOXSD
IEPH9VM01AM1JQ
ILANSYW03GMVYO
ILANSYW03GV3QM
IEPH9VM01EI3AT
IEPH9VM01A8N27IEPH9VM01A3WND
IEPH9VM01CCRJA
ILANSYW03GGU5B
IEPH9VM01ALGU8
IODIJDD03GD9JN
IODIJDD03G50K2
IODIJDD03GXOHH
IODIJDD03FT54TIODIJDD03FVLYY
ILANSYW03GFFBZ
IEPH9VM01ECSO8
ILANSYW03GQO6BIODIJDD03F42DG
ILANSYW03G3PVN
ILANSYW03F0V9I
ILANSYW03HCR8O
ILANSYW03G16FX
ILANSYW03GQ6JM
ILANSYW03F6T9Z
ILANSYW03GLUHW
ILANSYW03F5HDQ
ILANSYW03GD7XD
ILANSYW03G0OSY
ILANSYW03GZ7F9
ILANSYW03FZ3D6
ILANSYW03G
FZ1J
IEPH9VM
01D73ZR
ILANSYW
03HEN
0C
ILANSYW
03HG
8DW
ILANSY
W03G
IAIE
ILANSYW
03HEG
SQ
ILANSYW03FSH
IF
ILANSYW03HBRC2
ILANSYW03G
AK9G
ILANSYW03FZ59M
ILANSYW03GJ5V6
ILANSYW03GRAK2
ILANSYW03GKHYO
ILANSYW03GC5VU
ILANSYW03HEM6E
ILANSYW03F9Z7Y
ILANSYW03G43MX
ILANSYW03GPMHE
ILANSYW03FZNG4
ILANSYW03GUZUJ
IODIJDD03G6NZYILANSYW03FPJJG
IEPH9VM01C4H16
IODIJDD03GM3XGIEPH9VM01C8X77
IODIJDD03F43LFIODIJDD03G4YJS
IODIJDD03FWI82
ILANSYW03GSAH8IODIJDD03GF3PB
IODIJDD03GM9YO
ILANSYW03FTX8E
IEP
H9
VM
01E
EP
JQ
IEP
H9
VM
01
DT
K2
YIL
AN
SY
W03
GD
CH
Q
IOD
IJD
D0
3H
AU
M0
ILA
NS
YW
03G
YT
6W
ILA
NS
YW
03F
1LC
ZIE
PH
9V
M01C
VW
JR
IEP
H9V
M01E
V5P
VIE
PH
9V
M01D
B7S
FIL
AN
SY
W03G
9JV
H
IEP
H9V
M01
EN
QU
F
ILA
NS
YW
03H
D58Z
ILA
NS
YW
03H
HJIY
ILA
NS
YW
03G
87D
W
IOD
IJD
D03F
X31F
IOD
IJD
D03G
882V
IOD
IJD
D03
FS
Z8A
IOD
IJD
D03
HC
6QH
IOD
IJD
D03G
OR
BH
IODIJ
DD03G
9Q4S
IODIJ
DD03F6W
8Y
IODIJ
DD03
G7W
PM
IEPH
9VM
01C2V
B1
IODIJDD03
GVU5C
IODIJ
DD03
GQ
ETE
IODIJ
DD03
GM
I0U
IODIJ
DD03
GR5H
M
IODIJ
DD03
G7V
93
IODIJ
DD03
F1O
G3
IOD
IJD
D03
HG
4RR
IEPH
9VM
01CVG
3M
IODIJ
DD03GTB7V
IODIJ
DD03GUB89
IODIJ
DD03F3I8J
IODIJDD03GYFSI
IODIJDD03GS211
ILANSYW03F15WG
IODIJDD03FQIOPIO
DIJDD03GU6UX
ILANSYW03G7RJ2IODIJDD03GBLAD
IODIJDD03HG68B
IODIJD
D03HE9Q8
ILANSYW
03FJX4X
ILANSYW
03G8FTN
IODIJ
DD03GEW4X
IOD
IJD
D03
G1O
DY
IOD
IJD
D03
GW
BBO
IOD
IJD
D03
GQ
SE4
IOD
IJD
D03
G8R
7Z
IOD
IJD
D03F
5K
45
ILA
NS
YW
03G
AJG
D
ILA
NS
YW
03H
D2LG
IOD
IJD
D03G
0N
4C
IEP
H9V
M01C
WR
ZR
IOD
IJD
D03FM
5D
N
IOD
IJD
D03H
AV
38
ILA
NS
YW
03FN
UW
X
IEP
H9V
M01C
7JG
7
IOD
IJD
D03H
F77F
IEP
H9V
M01A
I6C
H
ILA
NS
YW
03G
9T
QX
Legend:
Dataset HM
0
6
12
20
4913
9806
14700
Color ranges:
Proteobacteria
Other
Actinobacteria
Firmicutes
Bacteroidetes
Acidobacteria
IOD
IJD
D03F
W39G
IOD
IJD
D03G
OLT
Z
IOD
IJD
D03G
8V
L1
IOD
IJD
D03G
D7X
4
IOD
IJD
D03H
FLA
U
IOD
IJD
D03G
5Y
2J
IOD
IJD
D0
3G
JT
2C
IOD
IJD
D03
FK
DF
R
IOD
IJD
D03G
LG
F9
IOD
IJD
D0
3F
VN
F3
IOD
IJD
D0
3G
D4
4A
IOD
IJD
D0
3F
VT
4A
IOD
I JD
D0
3F
P0
L7
ILA
NS
YW
03
GG
XS
B
I OD
IJD
D0
3G
KP
SP
IOD
IJD
D0
3F
8D
XL
IOD
IJD
D0
3F
7W
U0
IEP
H9V
M01C
RR
N1
IOD
IJD
D03F
ZB
4S
IOD
IJD
D03F
O9B
X
IOD
IJD
D03F
UU
US
IOD
IJD
D03G
YS
FU
IOD
IJD
D03H
AN
XK
IOD
IJD
D03G
7P
VX
IOD
IJDD
03F
U6W
0
IOD
IJDD
03F
UX
DF
IOD
IJD
D03G
KC
GP
IEP
H9V
M01C
6T
0M
ILA
NS
YW
03G
XA
31
IOD
IJD
D03G
NQ
R0
ILA
NS
YW
03G
4F
JA
ILA
NS
YW
03H
E5M
I
IOD
IJD
D03H
FQ
8I
IEP
H9V
M01E
BC
Q1
IOD
IJD
D03G
KT
1W
IOD
IJD
D03H
AM
HE
IEP
H9V
M01B
BG
Z2
IOD
IJD
D03F
T3Y
9
IOD
IJD
D0
3H
DJT
2
IOD
IJD
D03
G5
CD
1
IOD
IJD
D0
3G
577O
IOD
IJD
D0
3G
I1Q
S
IOD
IJD
D03
G7
HO
Y
IOD
IJD
D0
3G
CM
WS
IOD
IJD
D03G
QP
DV
IOD
IJD
D03G
UT
PF
IOD
IJD
D0
3G
C8
4D
ILA
NS
YW
03FX
W0J
ILA
NS
YW
03G
P0D
V
IOD
IJD
D03G
46U
D
IOD
IJD
D03G
06G
3
IOD
IJD
D03FTLM
K
IOD
IJD
D03G
MTM
7
IOD
IJD
D03
HA
VC
D
IOD
IJD
D03
FZZPL
IOD
IJD
D03
G8Q
I7
IOD
IJD
D03G
570C
IOD
IJD
D03G
NR
07
IODIJ
DD03
GPK03
IODIJ
DD03
HEK9F
IODIJ
DD03
F72G
A
IODIJ
DD03
G4W
1U
IODIJ
DD
03G
AK3MIO
DIJ
DD03
FSTV
G
IODIJ
DD03
GX8E
X
IODIJ
DD03
GM
HRS
IODIJ
DD03F
UG
04
IODIJ
DD03
GRN94
IOD
IJD
D03
GLN
JG
IOD
IJD
D03
HC
OXZ
IOD
IJD
D03
HG
FW
K
IOD
IJD
D03
GC
M50
IOD
IJD
D0
3F
0T
5O
IOD
IJD
D03
GK
N0S
ILA
NS
YW
03
GR
QU
C
IODIJD
D03G
ZKVR
IEPH9V
M01A
P392
IODIJD
D03G
BLJ8
IOD
IJDD
03G
9W
0B
IOD
IJDD
03G
AC
US
IOD
IJDD
03FO
SS
M
IOD
IJDD
03G
G4LG
IOD
IJDD
03F4N
MT
IOD
IJDD
03FS
52R
IOD
IJDD
03H
AC
LG
IOD
IJDD
03G
NX
QT
IOD
IJDD
03G
OO
LY
IOD
IJDD
03GA2Z
9
IOD
IJDD
03GC
DN
T
IOD
IJDD
03GG
SD
5
IOD
IJDD
03G0S
VI
IOD
IJDD
03GO
4QL
IODIJD
D03G
OK3X
IODIJD
D03G
90CQ
IODIJD
D03G
YX2S
IOD
IJDD
03GO
9BS
IODIJ
DD03H
C0FH
IODIJ
DD03FLIDO
IODIJD
D03FMEKW
ILANSYW
03GM
4CZ
ILAN
SYW03G
20IE
IEPH
9VM
01B62Z
F
ILA
NS
YW
03H
HO
6L
IEP
H9V
M01
DM
SY
4
ILAN
SYW
03H
CF9Y
ILAN
SYW
03F3Q
MV
ILAN
SYW
03F74W
L
IEP
H9V
M01A
5TK
Y
ILA
NS
YW
03F
5K
AF
IEP
H9V
M01D
JPR
K
IEP
H9V
M01E
CJI3
IEP
H9V
M01E
JVV
6
IEP
H9V
M01D
NH
JQ
IEP
H9V
M01E
CY
7C
ILA
NS
YW
03G
CR
2C
IEP
H9V
M01D
B64Y
IEP
H9V
M01B
6I2
2IE
PH
9V
M01D
WP
RL
IEP
H9V
M01B
EJO
TIE
PH
9V
M01C
27P
UIE
PH
9V
M01B
SB
5N
ILA
NS
YW
03
GQ
H5
AIE
PH
9V
M0
1D
ZE
2W
IEP
H9
VM
01E
ZN
W6
IEP
H9V
M01C
802I
IEP
H9
VM
01B
33E
L
ILA
NS
YW
03G
MW
CP
ILA
NS
YW
03FQ
Q8T
ILA
NS
YW
03H
GIZ
M
IEP
H9V
M01B
TS
KT
IEP
H9V
M01B
UV
7U
IEP
H9V
M01B
Z1M
N
IEP
H9V
M01D
IHU
0
ILAN
SYW
03FW
L9P
ILAN
SYW
03HD
R0Z
IEPH9V
M01C
6W5S
IEPH9V
M01D
9DNK
IEPH9V
M01C
P9K
1
ILAN
SYW
03GW
E9S
ILANSYW
03GPYAH
ILANSYW03FZO
FE
ILANSYW03G
KJTA
ILANSYW
03HB55O
ILANSYW
03GDZIZ
ILANSYW
03HE047
ILANSYW
03GM
3FBILANSYW03G
JCGN
ILANSYW
03G7C
UA
ILANSYW
03HBA8Y
ILANSYW03GC367
IODIJDD03GSW28
IODIJDD03FUAVU
IODIJDD03G7C0M
IODIJDD03GCDRPIODIJDD03G8F2Q
IODIJDD03HDG6F
IODIJDD03GJJGV
IODIJDD03F7W1E
IODIJDD03F771I
IODIJDD03GXZV1
IODIJDD03F3RJ5
ILANSYW03F6Q1O
ILANSYW03GX9GS
ILANSYW03G2X8B
ILANSYW03FX3NTILANSYW03G7HKA
IEPH9VM01EOSLJILANSYW03GDT24
ILANSYW03GZHSJILANSYW03FW4HCILANSYW03GSMGX
IEPH9VM01CXRQJ
ILANSYW03F0MTUIODIJDD03GZRJD
IODIJDD03GBWGL
IODIJDD03HENGG
IODIJDD03GC2D1
IODIJDD03HCZXSIODIJDD03F3GXE
IODIJDD03F9QQZ
ILANSYW03G3YPX
IODIJDD03FKDPIIODIJDD03GLHAW
IODIJDD03G2XVY
IODIJDD03G88PTIODIJDD03GK581
IODIJDD03FNCLN
IODIJDD03F0SMX
IODIJDD03F0OW8
IODIJDD03G7MWN
IODIJDD03FT36L
IODIJDD03G0SUS
IODIJDD03GC6ZX
IODIJDD03GSL2O
IODIJDD03FTU44
ILANSYW03GPLIM
ILANSYW03F2C38
IODIJDD03GCOUI
IODIJDD03GEBX6IODIJDD03FKGWIILANSYW03GL4J0IODIJDD03GS13V
IODIJDD03GQFMV
IODIJDD03GJ3L5IODIJDD03GI6JK
IODIJDD03G2J77
IODIJDD03G0TDW
IEPH9VM01DWFIYIEPH9VM01AKOEAIEPH9VM01EGIEI
ILANSYW03HDQ2J
IODIJDD03G1EGR
IODIJDD03F3MH1
IODIJDD03GSADN
IODIJDD03GT43M
IODIJDD03FUYTF
ILANSYW03GBUWZ
IODIJD
D03HDW9P
ILANSYW
03HD0LE
ILANSYW03GKVW
DIO
DIJDD03FZEHF
IODIJDD03GLI5X
IODIJDD03GE65U
IODIJDD03GRPMO
ILANSYW03F1SV7
IODIJDD03GWFKG
IODIJDD03GUESG
IODIJDD03G11ZUIODIJDD03FVHTW
IODIJDD03GK0NP
IODIJDD03GIJGL
IODIJDD03HB70ZIODIJDD03GLJ9Z
IEPH9VM01BOXSD
IEPH9VM01AM1JQ
ILANSYW03GMVYO
ILANSYW03GV3QM
IEPH9VM01EI3AT
IEPH9VM01A8N27IEPH9VM01A3WND
IEPH9VM01CCRJA
ILANSYW03GGU5B
IEPH9VM01ALGU8
IODIJDD03GD9JN
IODIJDD03G50K2
IODIJDD03GXOHH
IODIJDD03FT54TIODIJDD03FVLYY
ILANSYW03GFFBZ
IEPH9VM01ECSO8
ILANSYW03GQO6BIODIJDD03F42DG
ILANSYW03G3PVN
ILANSYW03F0V9I
ILANSYW03HCR8O
ILANSYW03G16FX
ILANSYW03GQ6JM
ILANSYW03F6T9Z
ILANSYW03GLUHW
ILANSYW03F5HDQ
ILANSYW03GD7XD
ILANSYW03G0OSY
ILANSYW03GZ7F9
ILANSYW03FZ3D6
ILANSYW03G
FZ1J
IEPH9VM
01D73ZR
ILANSYW
03HEN
0C
ILANSYW
03HG
8DW
ILANSY
W03G
IAIE
ILANSYW
03HEG
SQ
ILANSYW03FSH
IF
ILANSYW03HBRC2
ILANSYW03G
AK9G
ILANSYW03FZ59M
ILANSYW03GJ5V6
ILANSYW03GRAK2
ILANSYW03GKHYO
ILANSYW03GC5VU
ILANSYW03HEM6E
ILANSYW03F9Z7Y
ILANSYW03G43MX
ILANSYW03GPMHE
ILANSYW03FZNG4
ILANSYW03GUZUJ
IODIJDD03G6NZYILANSYW03FPJJG
IEPH9VM01C4H16
IODIJDD03GM3XGIEPH9VM01C8X77
IODIJDD03F43LFIODIJDD03G4YJS
IODIJDD03FWI82
ILANSYW03GSAH8IODIJDD03GF3PB
IODIJDD03GM9YO
ILANSYW03FTX8E
IEP
H9
VM
01E
EP
JQ
IEP
H9
VM
01
DT
K2
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AN
SY
W03
GD
CH
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D0
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M0
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03G
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6W
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03F
1LC
ZIE
PH
9V
M01C
VW
JR
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H9V
M01E
V5P
VIE
PH
9V
M01D
B7S
FIL
AN
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W03G
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IEP
H9V
M01
EN
QU
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ILA
NS
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03H
D58Z
ILA
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03H
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ILA
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03G
87D
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IOD
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D03F
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D03G
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D03
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D03G
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BH
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PM
IEPH
9VM
01C2V
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IODIJDD03
GVU5C
IODIJ
DD03
GQ
ETE
IODIJ
DD03
GM
I0U
IODIJ
DD03
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IODIJ
DD03
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93
IODIJ
DD03
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G3
IOD
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D03
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IEPH
9VM
01CVG
3M
IODIJ
DD03GTB7V
IODIJ
DD03GUB89
IODIJ
DD03F3I8J
IODIJDD03GYFSI
IODIJDD03GS211
ILANSYW03F15WG
IODIJDD03FQIOPIO
DIJDD03GU6UX
ILANSYW03G7RJ2IODIJDD03GBLAD
IODIJDD03HG68B
IODIJD
D03HE9Q8
ILANSYW
03FJX4X
ILANSYW
03G8FTN
IODIJ
DD03GEW4X
IOD
IJD
D03
G1O
DY
IOD
IJD
D03
GW
BBO
IOD
IJD
D03
GQ
SE4
IOD
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D03
G8R
7Z
IOD
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D03F
5K
45
ILA
NS
YW
03G
AJG
D
ILA
NS
YW
03H
D2LG
IOD
IJD
D03G
0N
4C
IEP
H9V
M01C
WR
ZR
IOD
IJD
D03FM
5D
N
IOD
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D03H
AV
38
ILA
NS
YW
03FN
UW
X
IEP
H9V
M01C
7JG
7
IOD
IJD
D03H
F77F
IEP
H9V
M01A
I6C
H
ILA
NS
YW
03G
9T
QX
ILMH
SAF
ISS
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Actinobacteria
Proteobacteria
Firmicutes
Cyanobacteria
Fusobacteria
Bacteroidetes
Other
Total Viable
ISS
Total Viable
SAF
Total Viable
ILMH • Phylogenetic tree showing the relationship in heatmap form of the shared OTU among the three sampling locations. Tree only displays the viable (PMA treated) OTUs.
Viable Microbial Burden-ATP Assay
Viable Bacterial Burden-qPCR
ILMH SAF
ISS
qPCR analysis
-Nature-Adam J Friedman, et al. 2013
-CDC
-Anesti, et al, 2005
Jet Propulsion Laboratory
California Institute of Technology
George C. Marshall Space Flight Center
University of North Dakota
Copyright 2014. All rights reserved.
