Microbiological research onboard the ISS and planetary protection with a special reference to dormancy problem

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Microbiological research onboard the ISS and
planetary protection with a special reference to
dormancy problem Natalia Novikova Institute
for Biomedical Problems, Russia
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THE MICROBIOLOGICAL FACTOR OF SPACE FLIGHT
Original microbiota of decorative-finish and
structural materials, gaseous and liquid
environments
Microbial contamination of interior and
equipment at the phases of assembly and
preflight preparation
Automicroflora of crew members
Contamination by microorganisms in the course
of loads delivery for space object
construction in near Earths orbit
Formation of microbial community in the internal
volume of space vehicle
Evolution of microbial community due to the
specifically changed environment of space
vehicle and cosmophysic factors
Devolvement of medical and technical
(technological) risks associated with the
existence of microorganisms in space vehicle
The system of preventive measures, scheduled
sanitary-hygienic operations, methods, means,
and technologies to counteract and mitigate
microbiological risks
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Hardware used for microbiological sampling of air
in the ISS
Refrigerator Thermostat Criogem-03
Ecosphere kit
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Hardware used for microbiological sampling of
internal surfaces and equipment in the ISS
View of the inside of the surface pipette kit
View of the outside of the surface pipette kit
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77species of microorganisms were found
Fungi
Bacteria
14 genera
11 genera
36 species
41 species
Including
Opportunistic pathogens BACTERIA Staphylococcus
aureus Streptococcus sp. Bacillus cereus
Opportunistic pathogens FUNGI Aspergillus
flavus Aspergillus niger Candida
parapsilosis Rhodoturula Rubra
FUNGI biodestructors Aspergillus niger
Aspergillus versicolor Penicillium
aurantiogriseum Cladosporium herbarum,
Cladosporium cladosporioides Ulocladium botrytis
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MICROBIOLOGICAL RISKS IN SPACE FLIGHT
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FUNGAL GROWTH IN THE INTERIOR OF
ORBITAL STATION SALYUT-7
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CONTAMINATION OF WINDOW BY MOLD FUNGI IN
LABORATORY EXPERIMENT
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GROWTH OF MOLD FUNGI ON THE
COMMUNICATION DEVICE WHITE AND BLACK TUBES
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GROWTH OF MOLD FUNGI ON THE COMMUNICATION DEVICE
INSULATION BLOCK
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Fungal Growth on the Fire Detecter
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Biocorrosion of the Fire Detecter Needle
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MATERIALS BIODEGRADATION
RUBBER
TITANIUM
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MATERIALS BIODEGRADATION
ALUMINIUM
ELECTRICAL TAPE
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DAMAGES of ALUMINIUM
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A SYSTEM OF MEASURES AIMED AT PROVIDING
MICROBIAL SAFETY OF THE ORBITAL STATION
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Problems

• Lately (14-17 basic expeditions) the quality a
  sanitary-microbiological conditions of FGB module
  has decreased. This includes exceeding normative
  parameters of microorganisms, mostly fungi,
  presence in FGB environment and on surfaces.

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Exceeding of standard on microorganisms
contents in FGB during ISS-15, 16 and ISS-17
flights
Air 98th day of the flight of ISS-16 and 197th
day of the flight of ISS-17 CFU/ m3 (standard
given in ISS MORD 50 260 bacterium 1000
CFU/m3, fungi 100 CFU /m3)
Surfaces 168th day of the flight of ISS-15 CFU
/ 100 cm2 (standard given in ISS MORD 50 260
bacterium 10,000 CFU/ 100 cm2, fungi 100 CFU
/100 cm2)
Sampling areas Fungi Fungi
FGB 242 1056
Tested surfaces Bacterium Fungi
FGB, behind the panel 230 1,5?102 1,2?105
FGB, on the panel 408 contamination area 2,8?104 4,0?104
FGB, on the panel 404 contamination area 8,5?104 5,0?101
SM, behind the panel 139 3,8?104 Non detectet
SM, niche 3,8?104 2,4?106
Contamination on the panel 408 FGB
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Reasons

• Unauthorized usage of FGB for washdowns and wet
  towels and clothes drying by crew members

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FGB Panel 406
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Problem solutions KIT FUNGISTAT
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Problem solutions Potok 150MK

• Apparently the onboard operation of the air
  sterilization device Potok 150MK was beneficial
  for the reduction of microbial air contamination
  of SM.
• In January 2009 the second Potok 150MK was
  delivered to ISS and was disposed in FGB.

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POTOK 150MK unit for International Space Station,
module Zvezda
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POTOK technologyAir sterilisation and fine
filtration
The main technical data

• - Efficiency of sterilization - 99100
• - Efficiency of filtration particle size 0,0110
  micron - up to 99
• - Weight 9 kg
• - Dimension 420322360 mm

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Inactivation through the combination of field and
current effects
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Mechanism of action of POTOK technology
Before Effects After
Total structural destruction (Saccharomyces cerevisiae)
Multiple perforation of membrane (Pseudomonas fluorescens)
Explosion of cytoplasm and cell membrane deformation (Micrococcus luteus)
Method Electronic microscopy / Cryofractography
ultra-thin sections
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The development of new means and methods for
providing materials antimicrobial resistance

• Biocides
• Application of various biocides as liquid means
  of cleansing has a number of disadvantages. They
  are
• - Large labor expenditures for the executive
• - Short-term effect
• - Necessity of periodic repetition of a
  cleansing
• - Poor efficiency in case of violation of the
  application technology when the part of the
  material remains untreated
• - Toxicity for human.

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GROWTH FUNGI ON A MATERIAL (after application by
a biocide)
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GROWTH FUNGI ON A MATERIAL (after application by
a biocide)
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New method for protection

• The most perspective model of protection is the
  creation of such material where the biocide is
  fixed with its surface and forms molecular layer.
• The advantages of this method are the following
• - the technological characteristics of the
  material in its volume do not change
• - high concentration of a biocide is formed on
  the surface
• - due to chemical bond with the surface of
  material the prolonged effect of protection is
  provided.

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EXPERIMENTAL CLIMATE CHAMBER
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Scheme of Experimental chamber

• 1 Thermostat
• 2 - Drum
• 3 Drum's slide rails
• 4 Holder with the specimens
• 5 Flat low freqiency electromagnetic radiator
• 6 Pin antennae of high frequency radiator
• 7 Temperature sensor
• 8 Humidity sensor
• 9 - Electrolyte bath (CAM)
• 10 Ventilator
• 11 Heater
• 13 -Support
• 14 Neutron and gamma sources
• 15 Radiation shielding
• 12 -Window

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Selection of material samples

• I. ALUMINUM FOIL AMG-6
• II. ORGANIC GLASS
• (Polymethylmethacrylate)

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ALUMINUM FOIL AMG-6
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ORGANIC GLASS (Polymethylmethacrylate)
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Result of the research

• Anti-microbial film coatings with different
  biocide concentration have been developed and
  prepared.
• On the basis of these complexes compositions have
  been derived and coatings have been formed on the
  substrates from aluminum allow and
  polymethylmetacrillate (PMMA).

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Space experiment Biorisk

• Petri dish with sistem microorganisms -
  structural material

Equipment for the Biorisk experiment
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• Microorganisms used in the
• Biorisk experiment

BACTERIA Bacillus pumilus Bacillus
licheniformis Bacillus subtilis
FUNGI Aspergillus versicolor Penicillium
expansum Cladosporium cladosporioides
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Cells of Bacillus subtilis
After flight
Before flight
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Hyphae of Aspergillus versicolor
After flight
Before flight
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Dynamics of P.expansum acid formation

• control before flight
• experiment after 7 months of exposure in outer
  space
•  

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Biological objects used in the 2-nd stage of
Biorisk experiment
BACTERIA Bacillus subtilis Bacillus subtilis Bacillus pumilus Bacillus licheniformis. FUNGI Aspergillus sydowii Aspergillus versicolor Penicillium aurantiogresium Penicillium expansum
ANIMALS Daphnia magna Daphnia pulicarias Streptocephalus torvicornis Artemia salina Eucypris species Polypedium vanderplanki Nothobranchius guenhteri SEEDS Brassica rapa Aabidopsis thalianaNaruna nijo
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BIORISK outer space experiment onboard Russian
segment of ISS
Germinated space seeds of Hordeum vulgare cv.
Haruna Nijo and plants of radish Raphanus sativus
strain Cherry Bomb, planted from space seeds
Biorisk-MSN canisters and their location on the
PIRS Docking compartment
Growth of Bacillus subtilis 24 after 13
month of exposure to outer space on the outer
side of ISS
Sleeping chironomid larvae Nemuri-usurika
rehydrated after 13 months in outer space
Reactivation of space and control Artemia
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Experiment EXPOSE-R (started 10.03.2009)
Containers , where assembled samples are placed
Assembled container with samples. The diameters
of the openings are from 7 to 10 mm. UV
irradiation passing from 0 to 100
General view of EXPOSE-R
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• Location of biological samples in the trays of
  EXPOSE-R

Pressurized Teflon packages 7?7 mm and 10?10 mm
with biological samples (spores of fungi, dried
embryos of lower crustaceans, dried chironomid
larvae, seeds of plant)
Trays with fixed Teflon packages containing
biological samples
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THE PROGRAM OF PHOBOS-GRUNT

• In October this year, in the frame of
  Phobos-Grunt program, Russian automatic
  spacecraft with sample return mission will be
  launched to Phobos, one of the moons of Mars. The
  main aim of this experiment is to return a soil
  sample from Phobos to Earth. At the same time,
  containers containing more than 60 different
  biological samples in latent forms will be
  installed in the same spacecraft for further
  returning to Earth, together with the Phobos soil
  sample. At the moment we conduct preparation of
  the biological samples for this flight.

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THE PROGRAM OF PHOBOS-GRUNT

• The goal of this experiment is
• Investigation of the survivability
• of resting stages of different organisms under
  conditions of long-duration space flight to
  solving different tasks related to planetary
  quarantine and astrobiology.

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THE PROGRAM OF PHOBOS-GRUNT MISSION PLANETARY
PROTECTION
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• Thank you
• for your attention!