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1 Organisms in Space Greg Leonard and Darren Hughes Mains Associates.

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Presentation on theme: "1 Organisms in Space Greg Leonard and Darren Hughes Mains Associates."— Presentation transcript:

1 1 Organisms in Space Greg Leonard and Darren Hughes Mains Associates

2 2 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, Laika became an international celebrity, with several countries, from Romania to Mongolia, celebrating the event with commemorative stamps.

3 3 Ham Chimpanzee January 31, 1961 Sam Rhesus monkey Two flights: 1959 & 1963 History - Primates in Space Enos Chimpanzee November 29, 1961 Two orbits

4 4 Neurolab 1998 First dedicated neurology research program in orbit ‘Felix’ AG1 (France) 1963 First cat in orbit Spacelab First reusable animal laboratory in orbit Biosat I 1966 First bacteria in orbit NASA/Mir First seed- to-seed growth of plants in orbit ‘Arabella’ Skylab First student experiment in orbit ‘Enos’ Mercury First primate in orbit ‘Laika’ Sputnik II 1957 First organism in orbit Timeline: Key Milestones (1) John Glenn Mercury First American in orbit Yuri Gagarin Vostock I 1961 First human in orbit Biosat II 1967 First seeds germinated in orbit Bion 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

5 5 Neurolab 1998 First dedicated neurology research program in orbit ‘Felix’ AG1 (France) 1963 First cat in orbit Spacelab First reusable animal laboratory in orbit Biosat I 1966 First bacteria in orbit NASA/Mir First seed- to-seed growth of plants in orbit ‘Arabella’ Skylab First student experiment in orbit ‘Enos’ Mercury First primate in orbit ‘Laika’ Sputnik II 1957 First organism in orbit Timeline: Key Milestones (2) John Glenn Mercury First American in orbit Yuri Gagarin Vostock I 1961 First human in orbit Biosat II 1967 First seeds germinated in orbit Bion 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

6 6 Some Organisms Studied in Space Bacteria Aeromonas proteolytica Bacillus mycoides Bacillus subtilis Bacillus thuringiensis Burkholderia cepacia Chaetomium globosum Deinococcus radiodurans Escherichia coli Nematospiroides dubius Rhodotorula rubra Salmonella typhimurium Trichophyton terrestre Invertebrates Acheta 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) Plants Aesculus 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) Vertebrates Canis 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) Birds Coturnix coturnix (Quail) Gallus gallus (Chicken) Aquatic species Arbacia 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)

7 7 Benefits of Studying Different Organisms Benefits to Space Exploration Risk mitigation Medical care Life support Benefits to Life on Earth Biology Medicine Technology Education

8 8 Human Studies in Space Bone Deterioration Muscle Atrophy Cardiovascular Deconditioning Immune Suppression Sleep Disturbances Balance Disorders

9 9 Why Not Just Study Humans in Space? Primary Reasons: Ethical Practical Biological Medical Logistical

10 10 Why Study Microbes in Space? Basic research Bioregenerative life support Nanotechnology

11 11 Why Grow Plants in Space? Basic research Food source Remove CO 2 Produce O 2 & water vapor Psychological benefits

12 12 Current and Future Directions Focus on cell and molecular biology Understanding of underlying mechanisms Use of “model” organisms Reference studies

13 13 Model Organisms Well-characterized Genetically sequenced Appropriate for space research

14 14 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 °C; Optimum 37°C SalinityTolerates low to moderate salinity Nutrients LB for bacteria: yeast extract, bacto-peptone, sodium chloride, water pH Growth optimum between pH Doubling rate 20 minutes to several hours Light Not required Aeration E. coli is facultative anaerobe, i.e. does not require O 2 but grows better in itspresence Wastes Gaseous (CO 2 ) and liquid (metabolites) Bacteria (E. coli) µm Growth Requirements

15 15 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 °C; Optimum 28°C SalinityYeast grows within a wide range of salt concentration Nutrients YPD : yeast extract, bacto-peptone, glucose, water pH Growth optimum between Doubling rate 1-4 hours Light Not required Aeration Does not require O 2 for growth, but grows better in its presence Wastes Gaseous (CO 2 ) and liquid (metabolites) Growth Requirements Saccharomyces cerevisiae (Yeast)

16 16 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 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 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 (CO 2 ), liquid (metabolites), and debris (dead worms) Growth Requirements Caenorhabditis elegans (Nematode)

17 17 Drosophila melanogaster (Fruit fly) Microfluidics Solid medium Sensors pH, oxygen, carbon dioxide,temperature Temperature °C with heat shock at 45°C. Nutrients YPD : yeast extract, bacto-peptone, glucose, water pH ~ optimum; range ~7.2 – 4.5 Doubling rate 1-4 hours Light Not required Aeration Active or passive aeration required Wastes Gaseous (CO 2 ) and liquid (metabolites) Growth Requirements

18 18 Microfluidics Water/nutrient delivery system Sensors pH, carbon dioxide, oxygen, ethylene, temperature, light Temperature °C; some protocols call for 15°C during dark cycle Humidity %; vegetative phase tolerant of a broad relative humidity range but above 855 can affect flowering and seed set Nutrients/waterConsistent 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 CompositionTypical air composition: 21% O 2, 78% N, 0.05% CO 2 Wastes O 2, ethylene Growth Requirements Arabidopsis thaliana (Brassica)

19 19 Microfluidics Liquid culture flowing above an adherent cell layer or cells growing in suspension in a liquid culture Sensors pH, CO 2, O 2, 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 SalinityA 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 Doubling rate 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. CO 2 levels must be kept at certain level; typically 5% Wastes Gaseous (CO 2 ) and liquid (metabolites) Growth Requirements Mammalian Cells

20 20 Rodents HabitatRodent cage: ventilated and kept free of contaminants from urine & feces Sensors O 2, CO 2, temperature, activity (video) Temperature18°C - 26°C Humidity % Food/WaterIrradiated food bars (rodent chow) with long shelf-life / automatic watering manifolds or water bottles Population Density6 rats or 10 mice per cage Light 8-10 hours/day exposure during circadian cycle VentilationControl O 2, CO 2, particulate contaminants, animal odors Wastes Urine, fecesmust be contained Housing & Husbandry Requirements

21 21 Ethical Use of Animals at NASA Bioethical Principles Respect for life Societal benefit Nonmalificence

22 22 Regulations and Oversight IACUC and federal regulations Scientific standards Agency oversight Public scrutiny

23 23 Medical Operations of Space Flight I Dr Arthur Arnold, Jr Kennedy Space Center Life into Space Space Life Sciences Experiments Eds. K Souza, G Etheridge & P. X. Callahan Model Organisms for Space Biology Research Dr Rita Briggs Lockheed Martin Fundamentals of Space Biology Eds M. Asashima & G.M. Malacinski (1990) Japan Scientific Societies Press & Springer-Verlag Early History of Space Biology and Medicine John P. Marbarger Acta Astronautica Vol. 43 No. 1-2 pp International Flight Experiments Database NASA Life Sciences Data Archive References


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