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Rutgers symposium on lunar settlements 3-8 June 2007 Rutgers University A simple differential production method of silicon utilizing organisms for future use in lunar settlements Satadal Das Peerless Hospital & B. K. Roy Research Centre Kolkata, India Silicon utilizing organisms are probably the fittest living creatures having a capacity of survival in extraterrestrial situations where they can tolerate more environmental stress and strain than their equals on Earth. One can also classify them according to their silicon utilizing capacity. Abundance of chemicals on earth and moon 50 45 40 35 30 % Earth 25 Moon 20 15 10 5 0 Oxygen Silicon Aluminium Iron Calcium Magnesium Others It is well known that organisms with high silicon content can survive in extremes of temperature, pressure and radiation. In fact, Reynolds described temperature tolerance of silicon compounds in living creatures as early as in 1893. Thus organisms with high silicon content can aptly be utilized within artificial environments in extraterrestrial situations. There are distinct Silicon accumulator plants like Cyperaceae, Graminae, Juncaceae and Moquiles spp. Organisms like marine phytoplanktons, marine brown algae, ‘horsetails’, foraminifera and porifera contain enough silicon, in the range of 60,000-4,37,000 mg per kg dry matter, and bacteria contain about 180 mg silicon per Kg dry matter. There is a long list of silicon utilizing organisms. PROTOZOA Chrysomonadida Silicoflagellida Heterochlorida Ebriida Lobosia Arcellinida, Arcella, Difflugia Gromiida PROTOZOA Radiolaria Porulosida Oculosida Centrohelida Desmothoracida SPONGES (PORIFERA) Hexactinellida Euplectella (Venus’s flower basket) Hyalonema ( Glass rope sponge) Pheronema Demospongia Cliona Poterion Pachychalina Spongilla ALGAE Division Class Order Class Class : Chrysophycophyta : Chrysophyceae (golden–brown algae) : Rhizochrysidales Chrysamoeba Ochromonas : Bacillariophyceae (yellow–green algae) Diatoms : Xanthophyceae (yellow–green algae) Vaucheria LICHENS –All variety, Crustose, Foliose, Frutiose. FUNGI Aspergillus Penicillium Alternaria Cladosporium PLANTS Dryland grasses such as oats and rye Wetland Grasses Bamboo e.g. Bambusa glaucesscens Chlorophytum comosum (Spider Plant) Anthurium scherzerianum (Flemingo Lily) Calathea makoyana (Peacock Plant) Aechmea fasciata (Silver Vase) Spathipyllum (Peace Lily) Nephrolepsis exaltata (Boston Fern) Asparagus seteceus (Asparagus Fern) Equisetum arvense (Horsetail) Bambusa glaucescens (Bamboo) Agave Americana (Century Plant) Chamaedorea elegans (Parlor Palm) Codiaeum variegatium (Croton) Howea forsteriana (Kentia Palm) Schefflera actinophylla (Umbrella Tree) Syngonium podophyllum (Arrowhead Plant) Hedera helix (Ivy) Cordyline terminalis (Ti plant) good luck plant Hedera helix (Tree Ivy, Pia) Hypoestes phyllostachya (Pink Splash) Gynura aurantiaca (Purple Passion) Ficus benjamina (Weeping Fig) Philodendron scandens (Philodendron) Acalypha pendula (Red-hot cat’s tail) Aglaonema commutatum (Chinese Evergreen) Cyperus alternifolius (Umbrella Sedge) Peperomia clusifolia (Baby Rubber Plant) Epipremnum aureum (Pothos) Dieffenbachia maculata (Dumb Cane) Dracaena deremensis (Dragon Tree) Dracaena marginata (Dragon Tree) Rice Oryza sativa Sugarcane Wheat Citrus Strawberry Cucumber Tomato Rose BACTERIA Almost all gram positive bacteria There are some similarities between carbon and silicon as they both belong to period IV of the periodic table. Although carbon compounds are abundantly found in living creatures on Earth and they are the basis of evolution of life on earth, there was at least a minor role of silicon compounds in the development of the primitive forms of life when the earth was quite inhospitable for the development of carbon based life. Trevors (1997) Bacterial evolution and silicon. Antonie Van Leeuwenhoek, 71(3):271-6. Silicon utilizing organisms when cultivated on medium prepared with carbon free constituents containing little nitrogen and phosphates they could grow better after repeated subcultures probably with the help of a trace amount of carry-over carbon during inoculation procedures. When silicon level was studied by electron prove microanalyser after thorough washing steps we find that silicon in cells grown in carbon free silicate medium was 24.9% while when they were on conventional carbon based medium they contain only 0.84% silicon. In a series of studies by us we find that many gram-positive bacteria and fungi can grow on silicate medium prepared with carbon free chemicals. In almost all cases initial growth was earlier on silicate medium, however, further growth was not good on carbon- free silicate medium. Das et al (1992) Metabolism of silicon as a probable pathogenecity factor for Mycobacterium and Nocardia Sp. Indian J. Medical Research (A) 95,59 – 65. Das S (1995) “ Silicon utilization” – an important pathogenecity marker of Mycobacterium tuberculosis. The Japanese J. Clinical Pathology, 43 (Supple.), 261. Das et al (2000) Role of silicon in modulating the internal morphology and growth of Mycobacterium tuberculosis. Indian J. Tuberculosis. 47: 2000, 87-91. Organisms (Gram positive bacteria can grow on carbonfree silicate medium) Mycobacterium marinum Average no. of days required for appearance of growth on carbon free silicate medium Average no. of days required for appearance of growth on carbon-based routine medium 1 1 M.scrofulaceum 3 10 M. flavescens 3 5 M. gordonae 3 3 M. avium 3 10 M. intracellulare 10 10 M. terrae 5 5 M. triviale 5 5 M. xenopi 10 12 M. fortuitum 1 1 M. smegmatis 2 1 M. tuberculosis 3 7 Bacillus subtilis 1 1 B. pumilus 1 1 Lactobacillus casei 1 1 Streptomyces rimosus 5 1 S. venezuale 7 1 Nocardia asteroides 3 2 N. braziliensis 3 1 N. caviae 3 1 Penicillium notatum 1 1 Aspergillus spp. 1 1 Rhizopus spp. 10 1 Trochophyton rubrum 3 1 T. violaceum 3 1 T. tonsurans 3 1 T. mentagrophytes 3 1 Fungi when grown on carbon free medium they produced peculiar morphological patterns which are hitherto unknown to us. Penicillium spp. Aspergillus spp. Aspergillus spp. Mucor spp. Penicillium spp. Epidermophyton spp. Trichophyton spp. Streptomyces spp. Epidermophyton spp. Streptomyces spp. Silicon utilizing microorganisms can grow in anaerobic condition. They can tolerate different types of radiations. It was found that although there are some metabolic changes in silicon utilizing microorganisms in radiation, its gives a positive impact on the nutritional quality owing to reduction of C:P ratio. Commercial gardening experiment in international space stations indicated that seed to seed life cycle is possible in space. Plants may help in bioregenerative life support system to perform chemistry of life support. Plants not only release precious oxygen but they also help in recycle drinking water. Microgravity situation may induce less lignin formation in plants but this will not prevent growth of these organisms Si li c o in g na m r fo nd T i ta n i u m fo r T er r a It was also found that when titanium is present the growth of silicon utilizing organisms were more on solid medium while the growth was less in liquid medium. This creates an unique opportunity on lunar surface where both silicon and titanium are present. Silicon utilizing organisms can thrive in sodium metasilicate (SM) solution as high as up to 4% concentration. To confine common silicon utilizing organisms from the environment for future use in lunar settlements one has to prepare SM solutions of four different concentrations- 0.5%, 1%, 2% and 4%. After preparation of such solutions in plastic containers one has to keep them in a greenhouse for as long as 5 years. Different varieties of organisms will grow in different concentrationsfrom a light green color growth in 0.5% SM solution, yellow color growth in 1% SM solution, orange color growth in 2% SM solution and a scanty whitish color growth in 4% SM solution. Besides many unknown microorganisms, algae are present in every solution but are of different kinds. Diatoms of diverse varieties are found in profound numbers in 0.5% and 2% SM solutions; plenty unknown acid-fast bacilli are also found in 1% SM solution Growth in 0.5% Silicate Solution Growth in 2% Silicate Solution Algal Growth in Control and 0.5% Silicate Solution Control 0.5% silicate Algal Growth in 1.0% and 2.0% Silicate Solutions 1.0% silicate 2.0% silicate Diatoms in 0.5% and 2.0% Silicate Solutions 0.5% silicate 2.0% silicate Anaerobic Growth Mainly in 0.5% and 1.0% Silicate Solution Control 0.5% 1.0% 2.0% 4.0% Unidentified Anaerobic Bacteria in Silicate Solution Unidentified Acid-fast Bacillary Growth in 1% Silicate Solution Fungal Growth in Control, 0.5%, 1.0%, 2.0%, 4.0% Silicate Solutions Scanty Growth of Unknown Microorganisms in 4% Silicate Solution Control Silicate 0.5% Silicate 1.0% Silicate 2.0% Phytoplankton other than diatoms 1.00 0.75 0.25 Diatoms 1.00 4.00 (Macro) 1.00 4.00 (Micro) 0.25 Gram positive bacteria 1.00 1.00 2.00 0.50 0.25 Coliform 1.00 0.75 0.60 0.42 0.12 0.25 Silicate 4.0% 0.12 Control Silicate 0.5% ─ ─ Anaerobic bacteria 1.00 4.00 Biofilms with green algae 1.00 Rhizopus Acid-fast bacilli Main fungi Silicate 1.0% Silicate 2.0% Silicate 4.0% ─ ─ 4.00 1.00 ─ 0.75 0.25 ─ ─ Aspergi llus Aspergi llus Aspergi llus ─ Plenty Control Silicate 0.5% Nitrate 1.00 1.22 Sulfate 1.00 1.53 Chloride 1.00 Iron 1.00 Silicate 1.0% Silicate 2.0% Silicate 4.0% 1.72 1.55 1.58 1.42 1.65 0.96 0.94 1.12 4.06 1.98 1.18 3.78 0.32 1.17 pH changes in Silicate solutions after Growth of Silicon-utilising Microorganisms 14 12 10 8 pH 6 4 2 0 Control Silicate 0.5% Silicate 1.0% Silicate 2.0% Silicate 4.0% Phytoplanktons in Different Silicate Solutions 80 70 60 % 50 Green algae Brown algae 40 Blue green algae Red algae 30 Relative diatom masses 20 10 0 Control Silicate 0.5% Silicate 1.0% Silicate 2.0% Silicate 4.0% Chemical Changes in Silicate Solutions after Growth of Silicon-utilising Microorganisms 800 700 600 Chloride 500 mg/L Sulfate Nitrate -Nitrogen 400 Iron 300 200 100 0 Control Silicate 0.5% Silicate 1.0% Silicate 2.0% Silicate 4.0% The south pole for our primary lunar settlement A simple protocol may be followed to use these silicate-utilizing organisms in lunar settlements. After providing minimum essential requirements for life in lunar extraterrestrial situation, these organisms may be utilized. Otherwise the protocol may be followed directly on a lunar crater to allow the organisms to find out a suitable zone for their growth. Lunar Crater Protocol : Step 1 : Microterraforming on moon In the initial venture antibiosis between various species should be prevented. Thus phytoplankton should be used before zooplanktons. Diatoms of Eueurytherm variety of Nitzschia and Chaetoceros group may be selected initially. Then golden algae grown in 2% and then algae grown in 0.5%SM solutions may be scattered to boost up the algal inhabitants. Other siliconutilizing algae Silicon-utilizing bacteria Eu-eurytherm siliconutilizing algae Diatoms Step 1a : Eu-eurytherm phase 3-12 months Nitzschia Subcurvata N. Curta N. Cylindrus N. Prolongatoides N. Pneudonana Chaetoceros Dichaeta C. Neglectus Step 1b : High silicon utilizing algal phase 3-12 months Algae grown in 2.0% silicate Step 1c : Low silicon utilizing algal phase 3-12 months Algae grown in 0.5% silicate Step 1d : Lichens and gram-positive bacterial phase 3-12 months Sub cultivations even blind passage may be done if necessary for 5-10 times during extending steps. This is because active and passive dispersal mechanism will be less on lunar surface Step 2 : Macroterraforming of moon Important silicon utilizing plants (specific silicon utilizing strains) like horsetails, grasses, lilies, silver vase, spider plant and following that organisms (only extremophile variety) like rotifers, tardigrades, nematodes, protozoa, fungi and other bacteria may be added which will live in close association of small silicon utilizing plants and this process may continue. Dracaena deremensis (dragon tree) Giant Equisetum arvense (horsetail) Anthurium scherzerianum (Flemingo lily) Calathea makoyana (peacock plant) Aglaonema commutatum (Chinese evergreen) Cordyline terminalis (Ti plant) good luck plant Chlorophytum comosum (spider plant) Step 2a : High Silicon metabolizing plants phase 1-5 years Dryland grasses such as oats and rye Bamboo e.g. Bambusa Glaucesscens Chlorophytum comosum (Spider Plant) Anthurium scherzerianum (Flemingo Lily) Calathea makoyana (Peacock Plant) Aechmea fasciata (Silver Vase) Spathipyllum ( Peace Lily) Step 2a : Equisetum arvense (Horsetail) Schefflera actinophylla (Umbrella Tree) Hedera helix (Ivy) Cordyline terminalis (Ti plant) good luck plant Dracaena deremensis (Dragon tree) Dracaena marginata (Dragon tree) Step 2b : Silicon accumulator plant phase – continued phase in close association of all previous organisms Rice Oryza sativa Sugarcane Wheat Citrus Strawberry Cucumber Tomato Rose etc. etc. Step 2c : Introduction of rotifers, tardigrades, nematodes, protozoa. Artificial support protocol : In this protocol silicon utilizing organisms may be used to support growth of non silicon-utilizing organisms and to produce a biosphere in artificial support situations.As it is not practicable to carry all essential nutrients for lunar settlements creation of such biosphere is essential for future survival of inhabitants in lunar settlements. Regolith containing top Iron frame with thick glasses inside the outer border of regolith top Solar energy lights may provide occasional exposure in long darkness Welcome to the Moon