1. 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

2. 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.

3. Abundance of chemicals on earth and moon5 10 15 20 25 30 35 40 45 50
Oxygen
Silicon
Aluminium
Iron
Calcium
Magnesium
Others %
Earth
Moon

4. 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 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.

5. There is a long list of silicon utilizing organisms.
PROTOZOA
Chrysomonadida
Silicoflagellida
Heterochlorida
Ebriida
Lobosia
Arcellinida, Arcella, Difflugia
Gromiida

6. PROTOZOA
Radiolaria
Porulosida
Oculosida
Centrohelida
Desmothoracida

7. SPONGES (PORIFERA) Hexactinellida Euplectella (Venus’s flower basket)
Hexactinellida
Euplectella (Venus’s flower basket)
Hyalonema ( Glass rope sponge)
Pheronema
Demospongia
Cliona
Poterion
Pachychalina
Spongilla

8. ALGAE Division : Chrysophycophyta
Class : Chrysophyceae (golden–brown algae)
Order : Rhizochrysidales
Chrysamoeba
Ochromonas
Class : Bacillariophyceae (yellow–green algae)
Diatoms
Class : Xanthophyceae (yellow–green algae)
Vaucheria
LICHENS –All variety, Crustose, Foliose, Frutiose.

9. 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)

10. 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)

11. 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)

12. Rice Oryza sativa
Sugarcane
Wheat
Citrus
Strawberry
Cucumber
Tomato
Rose
BACTERIA
Almost all gram positive bacteria

13. 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.

14. 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.

15. 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.

17. 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,

18. (Gram positive bacteria can grow on carbon-free silicate medium)
Organisms
(Gram positive bacteria can grow on carbon-free silicate medium)
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
Mycobacterium marinum 1
M.scrofulaceum 3 10
M. flavescens 5
M. gordonae
M. avium
M. intracellulare
M. terrae

19. M. triviale 5
M. xenopi 10 12
M. fortuitum 1
M. smegmatis 2
M. tuberculosis 3 7
Bacillus subtilis
B. pumilus
Lactobacillus casei
Streptomyces rimosus
S. venezuale

20. 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

21. Fungi when grown on carbon free medium they produced peculiar morphological patterns which are hitherto unknown to us.

22. Streptomyces spp. Streptomyces spp.
Penicillium spp.
Aspergillus spp.
Aspergillus spp.
Mucor spp.
Penicillium spp.
Epidermophyton spp.
Epidermophyton spp.
Trichophyton spp.
Streptomyces spp.
Streptomyces spp.

23. Silicon utilizing microorganisms can grow in anaerobic condition
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.

24. 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

26. 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.

27. 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 concentrations- from 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.

28. 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

29. Growth in 0.5% Silicate Solution

30. Growth in 2% Silicate Solution

31. Algal Growth in Control and 0.5% Silicate Solution

32. Algal Growth in 1.0% and 2.0% Silicate Solutions

33. Diatoms in 0.5% and 2.0% Silicate Solutions

34. Anaerobic Growth Mainly in 0.5% and 1.0% Silicate Solution
Control
0.5%
1.0%
2.0%
4.0%

35. Unidentified Anaerobic Bacteria in Silicate Solution

36. Unidentified Acid-fast Bacillary Growth in 1% Silicate Solution

37. Fungal Growth in Control, 0.5%, 1.0%, 2.0%, 4.0% Silicate Solutions

38. Scanty Growth of Unknown Microorganisms in 4% Silicate Solution

39. Control
Silicate 0.5%
Silicate 1.0%
Silicate 2.0%
Silicate 4.0%
Phytoplankton other than diatoms
1.00
0.75
0.25
0.12
Diatoms
4.00 (Macro)
4.00 (Micro)
Gram positive bacteria
2.00
0.50
Coliform
0.60
0.42

40. Control
Silicate 0.5%
Silicate 1.0%
Silicate 2.0%
Silicate 4.0%
Acid-fast bacilli ─
Plenty
Anaerobic bacteria
1.00
4.00
Biofilms with green algae
0.75
0.25
Main fungi
Rhizopus
Aspergillus

41. Control
Silicate 0.5%
Silicate 1.0%
Silicate 2.0%
Silicate 4.0%
Nitrate
1.00
1.22
1.17
1.72
1.55
Sulfate
1.53
1.58
1.42
1.65
Chloride
0.96
0.94
1.12
4.06
Iron
1.98
1.18
3.78
0.32

42. pH changes in Silicate solutions after Growth of Silicon-utilising Microorganisms2 4 6 8 10 12 14
Control
Silicate
0.5%
Silicate 1.0%
2.0%
4.0% pH

43. Phytoplanktons in Different Silicate Solutions10 20 30 40 50 60 70 80
Control
Silicate 0.5%
Silicate 1.0%
Silicate 2.0%
Silicate 4.0% %
Green algae
Brown algae
Blue green algae
Red algae
Relative diatom masses

44. Chemical Changes in Silicate Solutions after Growth of Silicon-utilising Microorganisms
100
200
300
400
500
600
700
800
Control
Silicate 0.5%
Silicate 1.0%
Silicate 2.0%
Silicate 4.0%
mg/L
Chloride
Sulfate
Nitrate -Nitrogen
Iron

45. The south pole for our primary lunar settlement

46. 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.

47. 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 Eu-eurytherm 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.

48. Other silicon-utilizing algae
Silicon-utilizing bacteria
Eu-eurytherm silicon-utilizing algae
Diatoms

49. Step 1a : Eu-eurytherm phase 3-12 months
Nitzschia Subcurvata
N. Curta
N. Cylindrus
N. Prolongatoides
N. Pneudonana
Chaetoceros Dichaeta
C. Neglectus

50. 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

51. 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.

52. Dracaena deremensis (dragon tree)
Giant Equisetum
arvense (horsetail)
Anthurium scherzerianum (Flemingo lily)
Cordyline terminalis (Ti plant) good luck plant
Calathea makoyana (peacock plant)
Chlorophytum comosum (spider plant)
Aglaonema commutatum (Chinese evergreen)

53. 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)

54. 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)

55. 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.

56. 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.

57. 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

58. Thank You
Welcome to the Moon