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Organisms in Space

Greg Leonard and Darren HughesMains Associates

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History - Dogs in Space

A Russian stray dog named Laika was the first biological specimen to orbit the Earth. She flew aboard the Sputnik 2 spacecraft launched on November 3, 1959.

Laika became an international celebrity, with several countries, from Romania to Mongolia, celebrating the event with commemorative stamps.

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HamChimpanzeeJanuary 31, 1961

SamRhesus monkeyTwo flights:1959 & 1963

History - Primates in Space

EnosChimpanzeeNovember 29, 1961Two orbits

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Neurolab 1998

First dedicated neurology research program in orbit

‘Felix’AG1

(France)1963

First cat in orbit

Spacelab 31985

First reusable animal laboratory in orbit

Biosat I1966

First bacteria in orbit

NASA/Mir1995-98

First seed-to-seed growth of plants in orbit

‘Arabella’Skylab 3

1973

First student experiment in orbit

‘Enos’Mercury 5

1961

First primate in orbit

‘Laika’Sputnik II

1957

First organism in orbit

Timeline: Key Milestones (1)

John Glenn Mercury 61961 First American in orbit

Yuri GagarinVostock I 1961First human in orbit

Biosat II1967

First seeds germinated in orbit

Bion 11973

First of 11 unmanned Russian biological research capsules

Future milestones (ISS and beyond):

• First mammal born in space

• First biology experiments beyond Earth orbit

• First multi-generational mammalian studies in space

• First self-sustaining ecosystem in space

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Neurolab 1998

First dedicated neurology research program in orbit

‘Felix’AG1

(France)1963

First cat in orbit

Spacelab 31985

First reusable animal laboratory in orbit

Biosat I1966

First bacteria in orbit

NASA/Mir1995-98

First seed-to-seed growth of plants in orbit

‘Arabella’Skylab 3

1973

First student experiment in orbit

‘Enos’Mercury 5

1961

First primate in orbit

‘Laika’Sputnik II

1957

First organism in orbit

Timeline: Key Milestones (2)

John Glenn Mercury 61961 First American in orbit

Yuri GagarinVostock I 1961First human in orbit

Biosat II1967

First seeds germinated in orbit

Bion 11973

First of 11 unmanned Russian biological research capsules

Future milestones (ISS and beyond):

• First mammal born in space

• First biology experiments beyond Earth orbit

• First multi-generational mammalian studies in space

• First self-sustaining ecosystem in space

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Some Organisms Studied in SpaceBacteriaAeromonas proteolyticaBacillus mycoidesBacillus subtilisBacillus thuringiensisBurkholderia cepaciaChaetomium globosumDeinococcus radioduransEscherichia coliNematospiroides dubiusRhodotorula rubraSalmonella typhimuriumTrichophyton terrestre

InvertebratesAcheta domesticus (Cricket)Araneus diadematus (Spider)Biomphalaria glabrata (Snail)Caenorhabditis elegans (Nematode)Cynops pyrrhogaster (Newt)Drosophila melanogaster (Fruit fly)Habrobracon juglandis (Wasp)Manduca sexta (Tobacco hornworm)Pelomyxa carolinensis (Amoeba)Pothetria dispar (Gypsy moth)Tribolium confusum (Beetle)Trigonoscelis gigas (Beetle)

PlantsAesculus hippocastanum L. (Horse chestnut)Arabidopsis thaliana (Thale cress)Avena sativa (Oat)Brassica rapa (Field mustard)Capsicum annuum (Ornamental pepper)Ceratodon (Moss)Ceratopteris (Fern)Ceratophyllum demersum (Hornweed)Cucumis sativus (Cucumber)Dactylis glomerata L. (Orchard grass)Daucus carota (Carrot)Digitalis lanata (Foxglove)Digitalis purpurea L. (Foxglove)Elodea (Waterweed)Flammulina velutipes, Agaricales (Fungus)Glycine max (Soybean)Haplopappus gracilis (Haplopappus)Helianthus annuus L. (Sunflower)Hemerocallis (Daylily)Lepidium sativum (Garden cress)Linum usitatissimum (Flax)Lycoperscion esculentum (Tomato)Neurospora crassa (Fungus)Nicotiana tabacum (Tobacco)Oryza sativa (Rice)Physarum polycephalum (Slime mold)Pseudotsuga menziesii (Douglas fir)Pseudotsuga taeda (Loblolly pine)Saccharomyces cerevisiae (Yeast)Tradescantia (Spiderwort)Triticum aestivum (Wheat)Triticum vulgare (Wheat)Vigna radiata (Mung bean)Zea mays (Corn)

VertebratesCanis familiaris (Dog)Felix maniculata (Cat)Homo sapiens (Human)Macaca mulatta (Rhesus monkey)Macaca nemestrina (Pigtail macaque monkey)Mus musculus (Mouse)Oryctolagus cuniculus (Rabbit)Pan troglodytes (Chimpanzee)Perognathus longimembris (Pocket mouse)Rattus norvegicus (Rat)Saimiri sciureus (Squirrel monkey)Testudo horsfieldi Gray (Tortoise)

BirdsCoturnix coturnix (Quail)Gallus gallus (Chicken)

Aquatic speciesArbacia punctulata (Sea urchin)Aurelia aurita (Jellyfish)Fundulus heteroclitus (Killifish)Lytechinus pictus (Sea urchin)Opsanus tau (Toadfish)Oreochromis mossambicus (Cichlid fish)Oryzias latipes (Medaka fish)Rana catesbeiana (Bullfrog)Rana pipiens (Frog)Strongelocentrotus pupuratus (Sea urchin)Xenopus laevis (Frog)Xenopus laevis Daudin (South African toad)Xiphophorus helleri (Swordtail fish)

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Benefits of Studying Different Organisms

Benefits to Space Exploration

• Risk mitigation

• Medical care

• Life support

Benefits to Life on Earth

• Biology

• Medicine

• Technology

• Education

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Human Studies in Space

• Bone Deterioration

• Muscle Atrophy

• Cardiovascular Deconditioning

• Immune Suppression

• Sleep Disturbances

• Balance Disorders

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Why Not Just Study Humans in Space?

Primary Reasons:

• Ethical

• Practical

• Biological

• Medical

• Logistical

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Why Study Microbes in Space?

• Basic research

• Bioregenerative life support

• Nanotechnology

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Why Grow Plants in Space?

• Basic research

• Food source

• Remove CO2

• Produce O2 & water vapor

• Psychological benefits

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Current and Future Directions

• Focus on cell and molecular biology

• Understanding of underlying mechanisms

• Use of “model” organisms

• Reference studies

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Model Organisms

• Well-characterized

• Genetically sequenced

• Appropriate for space research

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Escherichia coli (Bacteria)

Microfluidics Liquid culture likely for flight, can be grown on solid medium

Sensors pH, oxygen, carbon dioxide,temperature

Temperature Will grow between 10 - 45°C; Optimum 37°C

Salinity Tolerates low to moderate salinity

Nutrients LB for bacteria: yeast extract, bacto-peptone, sodium chloride, water

pH Growth optimum between pH 5.5 - 8.0

Doubling rate 20 minutes to several hours

Light Not required

Aeration E. coli is facultative anaerobe, i.e. does not require O2 but grows better in its

presence

Wastes Gaseous (CO2) and liquid (metabolites)

Bacteria (E. coli) 0.5-1.5µm

Growth Requirements

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Yeast (S. cer.) 5-12µm

Microfluidics Liquid culture likely for flight, can be grown on solid medium

Sensors pH, oxygen, carbon dioxide,temperature, pressure

Temperature Will grow at temperatures between 3 - 40°C; Optimum 28°C

Salinity Yeast grows within a wide range of salt concentration

Nutrients YPD : yeast extract, bacto-peptone, glucose, water

pH Growth optimum between 5.0 - 6.5

Doubling rate 1-4 hours

Light Not required

Aeration Does not require O2 for growth, but grows better in its presence

Wastes Gaseous (CO2) and liquid (metabolites)

Growth Requirements

Saccharomyces cerevisiae (Yeast)

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Microfluidics Liquid culture or solid culture. Organism is ~ 1mm in length and grows in axenic defined liquid media or on solid media using bacteria as food

Sensors carbon dioxide, oxygen, temperature

Temperature 17°C optimum for growth; heat shock at 25°C; 30°C for > 20 hrs is lethal

Salinity 0.1 - 0.5 molar simple inorganic salts (NaCl, KHPO4), wide tolerance range

Nutrients Consumes dissolved nutrients in axenic liquid culture media, or on solid media consumes bacteria (E. coli).

pH Optimum pH 6.0; tolerates pH 3 - 9

Doubling rate 3-6 days to mature; temperature dependent

Light Generally not required

Aeration Chamber ventilation required for axenic liquid culture in gas-permeable opticell cartridges or for culture on solid media

Wastes Gaseous (CO2), liquid (metabolites), and debris (dead worms)

Growth Requirements

Caenorhabditis elegans (Nematode)

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Drosophila melanogaster (Fruit fly)

Microfluidics Solid medium

Sensors pH, oxygen, carbon dioxide,temperature

Temperature 22 - 30°C with heat shock at 45°C.

Nutrients YPD : yeast extract, bacto-peptone, glucose, water

pH ~ 6.5 - 5.0 optimum; range ~7.2 – 4.5

Doubling rate 1-4 hours

Light Not required

Aeration Active or passive aeration required

Wastes Gaseous (CO2) and liquid (metabolites)

Growth Requirements

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Microfluidics Water/nutrient delivery system

Sensors pH, carbon dioxide, oxygen, ethylene, temperature, light

Temperature 17 - 25°C; some protocols call for 15°C during dark cycle

Humidity 65 - 100%; vegetative phase tolerant of a broad relative

humidity range but above 855 can affect flowering and seed set

Nutrients/water Consistent with soil composition; well aerated soil required

Doubling rate 4-8 week growth cycle

Light 16hr light, 8 hr dark cycle; light intensity 250 u mol

Gas Composition Typical air composition: 21% O2, 78% N, 0.05% CO2

Wastes O2, ethylene

Growth Requirements

Arabidopsis thaliana (Brassica)

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Microfluidics Liquid culture flowing above an adherent cell layer or cells growing in suspension in a liquid culture

Sensors pH, CO2 , O2 , temperature, pressure, flow rate

Temperature Tolerate only a very narrow temperature range, typically between 37°C - 42°C. Optimal temperature is 37°C ± 0.5°C

Humidity ~ 80% in incubators

Salinity A variety of commercial buffered saline solutions used; most popular are Hank’s and Earle’s

Nutrients Different cell types require different media formulations, e.g. DMEM: glucose, L- glutamine, sodium pyruvate, phenol red. Between 5-20% fetal bovine serum or horse serum is a common supplement.

pH pH range between 6.8 - 7.2

Doubling rate 12 - 48 hours

Light Not required

Aeration Air, as oxygen source, must be added to culture in a way to avoid sheer stress to which mammalian cells are very sensitive. CO2 levels must be kept at certain level; typically 5%

Wastes Gaseous (CO2) and liquid (metabolites)

Growth Requirements

Mammalian Cells

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Rodents

Habitat Rodent cage: ventilated and kept free of contaminants from urine & feces

Sensors O2 , CO2 , temperature, activity (video)

Temperature 18°C - 26°C

Humidity 30 - 70%

Food/Water Irradiated food bars (rodent chow) with long shelf-life / automatic watering

manifolds or water bottles

Population Density 6 rats or 10 mice per cage

Light 8-10 hours/day exposure during circadian cycle

Ventilation Control O2 , CO2 , particulate contaminants, animal odors

Wastes Urine, feces must be contained

Housing & Husbandry Requirements

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Ethical Use of Animals at NASA

Bioethical Principles• Respect for life• Societal benefit• Nonmalificence

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Regulations and Oversight

• IACUC and federal regulations• Scientific standards• Agency oversight• Public scrutiny

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Medical Operations of Space Flight IDr Arthur Arnold, JrKennedy Space Center

Life into Space Space Life Sciences Experiments 1991-1998Eds. K Souza, G Etheridge & P. X. Callahan

Model Organisms for Space Biology Research Dr Rita BriggsLockheed Martin

Fundamentals of Space BiologyEds M. Asashima & G.M. Malacinski (1990)Japan Scientific Societies Press & Springer-Verlag

Early History of Space Biology and MedicineJohn P. MarbargerActa Astronautica Vol. 43 No. 1-2 pp 9-12 1998

International Flight Experiments Databasehttp://www.mainsgate.com/IFE/index.html

NASA Life Sciences Data Archivehttp://www.lsda.jsc.nasa.gov

References