Rutgers symposium on lunar settlements

3-8 June 2007Rutgers 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

0

5

10

15

20

25

30

35

40

45

50

Oxygen Silicon Aluminium Iron Calcium Magnesium Others

%

Earth

Moon

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

ALGAEDivision : 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.

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 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 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.

Streptomyces spp.

Aspergillus spp.

Penicillium spp.

Mucor spp.

Trichophyton spp.

Epidermophyton spp.

Streptomyces spp.

Epidermophyton spp.

Penicillium spp.

Aspergillus 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

n um f

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 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.

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

0.5%Control 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%

Silicate 4.0%

Phytoplankton other than diatoms

1.00 0.75 0.25 0.25 0.12

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

Control Silicate 0.5%

Silicate 1.0%

Silicate 2.0%

Silicate 4.0%

Acid-fast bacilli ─ ─ Plenty ─ ─

Anaerobic bacteria

1.00 4.00 4.00 1.00 ─

Biofilms with green algae

1.00 0.75 0.25 ─ ─

Main fungi Rhizopus Aspergillus

Aspergillus

Aspergillus

─

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 1.00 1.53 1.58 1.42 1.65

Chloride 1.00 0.96 0.94 1.12 4.06

Iron 1.00 1.98 1.18 3.78 0.32

pH changes in Silicate solutions after Growth of Silicon-utilising Microorganisms

0

2

4

6

8

10

12

14

Control Silicate 0.5%

Silicate1.0%

Silicate2.0%

Silicate4.0%

pH

Phytoplanktons in Different Silicate Solutions

0

10

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

Chemical Changes in Silicate Solutions after Growth of Silicon-utilising Microorganisms

0

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

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 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.

Diatoms

Silicon-utilizing bacteria

Other silicon-utilizing algae

Eu-eurytherm silicon-utilizing algae

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 monthsAlgae grown in 2.0% silicateStep 1c : Low silicon utilizing algal phase 3-12 monthsAlgae grown in 0.5% silicateStep 1d : Lichens and gram-positive bacterial phase 3-12 monthsSub 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 moonImportant 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)

Cordyline terminalis (Ti plant) good luck plant

Chlorophytum comosum

(spider plant)

Anthurium scherzerianum (Flemingo lily)

Aglaonema commutatum (Chinese

evergreen)

Calathea makoyana (peacock 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 sativaSugarcaneWheatCitrusStrawberryCucumberTomatoRose 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.

Solar energy lights may provide occasional

exposure in long darkness

Regolith containing top

Iron frame with thick glasses inside the

outer border of regolith top

Welcome to the Moon