EXPERIMENT “BIOPHOBOS/ANABYOSIS”
IN PROJECT “PHOBOS-GRUNT”
О.I.Orlov1
V.R.Alekseev2
Е.А.Vorobyeva3
V.F.Galchenko4
Е.А.Deshevaya1
А.V.Zakharov5
М.А.Levinskikh1
О.V.Muravenko6
N.D.Novikova1
S.V.Poddubko1
N.А.Polykarpov1
V.N.Sychev1
1 - SSC RF – Institute of Biomedical Problems RAS, Moscow
2 - Zoological Institute RAS, St.Petersburg
3 - Soil Science Faculty, Moscow State University, Moscow
4 - Institute of Microbiology RAS, Moscow
5 - Space Research Institute RAS, Moscow
6 - Institute of Molecular Biology RAS, Moscow
LONG-TERM ASTROBIOLOGICAL RESEARCH
PROGRAM
Пути достижения поставленной цели
Long-term program of applied
science research and planned
experiments onboard RS ISS
MARS-500 Program
Satellite experiments
Experiments : Biorisk-MSV; Biorisk-
MSN; Expose-R; Aquarium; PlantsSeeds
Project “PHOBOS-GRUNT”
Experiment
“Biophobos/Anabiosis”
AIMS OF THE ASTROBIOLOGICAL RESEARCH
Elucidation of viability of resting forms of
organisms of different taxonomical groups
under conditions of spaceflight and continuous
interplanetary flights for solving problems of
planetary quarantine, astrobiology and biomedical problems of manned cosmonautics.
THE STUDY OF DORMANT FORMS OF VARIOUS
ORGANISMS UNDER SPACE FLIGHT CONDITIONS
To expose dormant forms of various organisms on the external
surface of the ISS, which include not only bacterial and fungal
spores but also seeds and dormant embryos of lower
crustaceans, as well as multi-cellular organism in the
cryptobiotic state.
Cooperation with:
INSIDE:
RASTENIYA-2
AQUARIUM
BRADOS
BIORISK - KM
BIORISC - MSV
OUTSIDE:
BIORISK – MSN
EXPOSE-R
Moscow State University, Moscow,
Russia;
Zoological Institute of RAS, St.
Petersburg, Russia;
Institute of microbiology of RAS,
Moscow, Russia;
St. Petersburg St. State University,
St. Petersburg, Russia;
Space Dynamic Laboratory, Utah
State University, Utah, USA;
Research Institute for
Bioresources, Okayama
University, Okayama, Japan;
National Institute of
Agrobiological Sciences, Ibaraki,
Japan.
EXPERIMENT “BIORISK-MSN-2”
HIGHER PLANTS
Air-dry seeds of mustard Brassica rapa, Improved Basic line
Air-dry seeds of mustard Brassica rapa, High Anthocyanin line
Air-dry seeds of mustard Brassica rapa, Anthocyaninless line
Air-dry seeds of mustard Brassica rapa, Rosette line
Air-dry seeds of mustard Brassica rapa, AstroPlants line
Air-dry seeds of mustard Brassica rapa, Somatic Variegation line
Air-dry seeds of mustard Brassica juncea, RG
Air-dry seeds of Arabidopsis thaliana, K-310 line
Air-dry seeds of Arabidopsis thaliana, Columbia line
Air-dry seeds of weed Nicandra physaloides
Air-dry seeds of tomatoes Lycopersicum esculentium, Micro-Tom
var.
Air-dry seeds of radishes Raphanus sativus, Cherry Bomb var.
Air-dry seeds of barley Hordeum vulgare, Haruna Nijo cv
Air-dry seeds of barley Hordeum vulgare, K-305 cv
Air-dry seeds of rice Oryza sativa, Rsp-5
Air-dry seeds of rice Oryza sativa, Rsp-7
VERTEBRATES
Dry eggs of fish Nothobranchius guenteri
INSECTS
Cryptobiotic larvae of chyronamide Polypedium vanderplankii
LOWER CRUSTACEANS
Dormant eggs of Daphnia magna
Dormant eggs of Daphnia pullicaria
Dormant eggs of Artemia salina
Dormant eggs of Streptocephalus torvicornis
Dormant eggs of Eucypris ornate
MOLDS
Spores of Aspergillus sedowii # 9-6
Spores of Aspergillus versicolor # 4-3-4
Spores of Penicillium aurantiogresium # 9-9
Spores of Penicillium expansum # 4-3-3
BACTERIA
Spores of Bacillus subtilis # 20
Spores of Bacillus licheniformis # 24
Spores of Bacillus subtilis # 25
Spores of Bacillus subtilis # 2335\105
Duration of the exposure:
The 1-st container – 13 months
The 2-nd container - 18 months
The 3-rd container – 31 months
EXPERIMENT “BIORISK-MSN” ONBOARD OF RUSSIAN
SEGMENT OF THE INTERNATIONAL SPACE STATION
“Biorisk-MSN” hardware and its location on
“Pirs” Docking Compartment
Growth of Bacillus subtilis # 24
strain after 13-months exposure
on the outer side of ISS
Germinated “space” seeds of barley Hordeum vulgare cv. Haruna
Nijo and plants of radish Raphanus sativus strain «Cherry Bomb»,
grown from “space” seeds
Larvae of chironomid
Polypedilum vanderplanki
revived after 13, 18 and 31
months of exposition on the
outer side of RS of ISS
Reactivation of “space” and control resting
eggs of Artemia salina
EXPERIMENT “EXPOSE-R” (Start March 10, 2009)
At present Russian scientists participate ESA
experiment EXPOSE-R onboard ISS. In
contrast to Biorsik-MSN experiment, a part
of biological samples in EXPOSE-R is being
exposed to solar UV.
BACTERIA
Group 18, subgroup Bacillaceae, genera Bacillus, species:
Bacillus subtilis-2335/105 ;
Bacillus subtilis-20 ;
Bacillus pumilus-25 ;
Bacillus licheniformis-24.
Gram-positive rods, 0,5-2,5 x 1,2-10 mkm, spore - 6 mkm
FUNGI
Taxon - Mitosporic fungi, genera Aspergillus, Penicillium, Geomyces species:
Aspergillus sydowi - 9-6;
Geomyces pannorum – 2241;
Aspergillus versicolor - 4-3-4;
Penicillium aurantiogresium – 9-9;
Penicillium expansum – 4-3-3.
Spore 3-10 mkm
ANIMALS
Dried encysted embryos (diameter of a single egg is 200 micron) Daphnia magna
belongs to family Daphniidae, order Daphniiformes, class Crustacea.
Dried encysted embryos (diameter of a single egg is 160 micron) Streptocephalus
torvicornis belongs to family Streptocephalidae, order Anostraca, class
Crustacea.
Dried encysted embryos (diameter of a single egg is 180-200 micron) Artemia salina
belongs to family Artemiidae, order Anostraca, class Branchiopoda,
subphylum: Crustacea.
Dried encysted embryos (diameter of a single egg is 180-200 micron) Eucypris
species belongs to family Cyprididae, order Ostracoda, class Crustacea.
Cryptobiotic dried larvae (length 5 mm   0,8 mm) of Polypedilum vanderplanki
belongs to family Chironomidae, order Diptera, class Insecta.
PLANTS
Seeds of Lycopersicum esculentum (Tomato Micro-Tom)
Seeds of Raphanus sativus (Radish Cherry Bomb)
Seeds of Arabidopsis thaliana line WT–2.
EXPERIMENT “EXPOSE-R” (Start March 10, 2009)
The biological objects in “EXPOSE-R” experiment are placed in plastic film bags 7x7
mm. The bags are fixed in the cells of special sets assembled in trays. EXPOSE-R
hardware contains three trays composed by 4 sets of the cells with biological samples.
The biological samples arranged in three levels, and those of the top level are exposed
to solar UV.
EXPERIMENT “BIOPHOBOS/ANABYOSIS”
For realization of biological research in “Phobos-Grunt” project, a
Workgroup chaired by academician RAS O.I. Orlov was organized by
decision of the section “Space biology and physiology” of RAS Council on
March 19, 2007. The workgroup consists of representatives of GSC IMBP
RAS, ISS RAS, ZIN RAS, MSU, IMB RAS, FSCP “SAO after S.A.
Lavochkin”. The workgroup has defined the main aims of biological
experiments (Experiment “BioPhobos/Anabiosis”) in “Phobos-Grunt”
project:
• investigation of survival of resting stages of bacteria, fungi, animals and
plants under conditions of interplanetary flights for estimation of
damaging effect of factors of space environment on cryptobiotic stages of
organisms and further development of technology of long-term
preservation of organisms in conditions of biological resting;
• investigation of biological, biochemical and molecular-genetic changes in
organisms after long-term spaceflight;
EXPERIMENT “BIOPHOBOS/ANABYOSIS”
• estimation of risk related to possibility of transformation of microorganisms
inhabiting the space object after their exposure to outer space with further
return inside of the space object.
• comparison of survival under conditions of the spaceflight of resting stages
with different level of survivability, including that of artificially induced be
mean of cultivation with biologically active substances, for developing new
biotechnological methods of investigations of resting stages for continuous
space transportation.
• comparison of survival of resting stages of organisms under space flight
conditions (BioPhobos/Anabiosis) and an orbital flight in control experiment
on low Earth orbit on the outer side of RS of ISS (experiment “Biorisk-MSN”).
EXPERIMENT “BIOPHOBOS/ANABYOSIS”
Род, вид
Штамм
Leptolyngbia sp.
Dp 6606
Methanosarcina lacustris sp.
nov. ZS (= DSM13486T), VKM B-2268
Methylosinus trichosporium
№ 34
Methylosinus sporium
№ 29
Methylocystis minimus
№ 28
Methylobacter chroococcum
№ 72
Deinococcus radiodurans
№ NP902
Artrobacter oxydens
№ Prm1
Pseudomonas aurefaciens
№ В1393
Bacillus subtilis
# 20
Bacillus licheniformis
#24
Bacillus pumilus
# 25
Bacillus subtilis
# 2335\105
Ferroplasma acidiphilum
gen. nov., sp. nov.
Acinetobacter sp.
К30А
Rhodococcus sp.
K-25
Sphingomonas sp.
К42
Acetobacterium tundrae sp.
nov. Z-4493 (=DSM1 9173T)
Micrococcus luteus
6/90-111
Aquaspirillum sp.
6/90-111/С505
Rudobacter spheroides
323
Streptomyces sp.
315
Spores of the
following
prokaryotes
(bacteria) are
included in the list
of experiment
“BioPhobos/
Anabiosis”
EXPERIMENT “BIOPHOBOS/ANABYOSIS”
Род, вид
Штамм
Ulocladium botrytis
16-12
Cladosporium cladosporioides
2-3
Aspergillus sydowii
9-6
Aspergillus versicolor
4-3-4
Penicillium aurantiogresium
9-9
Penicillium expansum
4-3-3
Aspergillus sydowii
ВКМ
Penicillium aurantiogresium
ВКМ
Aspergillus versicolor
ВКМ
Penicillium expansum
ВКМ
Aureobasidium pullulans
Viala&Boyer КБП-3847
Ulocladium botrytis
Preuss M-54 ВКМ F-4032D
Saccharomyces cerevisiae
Strain W303. ATCC 200060
Spores of the
following
eukaryotes (fungi)
are included in the
list of experiment
“BioPhobos/
Anabiosis”
EXPERIMENT “BIOPHOBOS/ANABYOSIS”
Animals
Род, вид
Порода
Artemia salina
Видовая форма
Streptocephalus torvicornis
Видовая форма
Eucypris sp.
Видовая форма
Daphnia magna
Видовая форма
Hemidiaptomus ingens
Видовая форма
Polypedilum vanderplankii
Видовая форма
Plants
Род, вид
Сорт
Arabidopsis thaliana
Видовая форма
Zingeria beibersteiniana
Видовая форма
Mesembryanthemum crystallinum
Видовая форма
Nicandra physaloides
Видовая форма
Resting forms of the
following higher
eukaryotes are included
in the list of experiment
“BioPhobos/Anabiosis”
Capsule with Phobos’ grunt and biological samples to be
returned to the Earth
For placing of the
biological objects in
two titanium
“Phobos-capsules”
with the size of  11
 35 mm, technology
similar to that of
“EXPOSE-R”
experiment will be
used.
2 “Phobos-capsule” with 122 (1010 mm) packs
with different biological objects
Thank you for your
attention