Feasibility of Cultivating Arthrospira platensis as a Food

Source Using Mars Regolith Simulant and Urine Simulant

for Mars Exploration and Colonization

Why Mars?

Planned Missions to Mars

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●

How Can We Survive on Mars? • Robert Zubrin ushered in

the idea of using indigenous resources on Mars to survive.

• Our future on Mars depends on our ability to use local resources and devise creative ways to exploit them to their benefit.

• The local Martian resources used in this study are urine and Mars regolith

What is Arthrospira platensis?

Spirulina• It’s NOT algae!• Cyanobacteria• Extremophile• Grows in water • Easy to process• Extremely

Nutritious

Benefits of Spirulina

Benefits of Spirulina

Why Urine?

Nutrients in Urine•

•

•

•

Why Mars Regolith?

Regolith CompositionMain trace elements that are needed are iron, zinc, and magnesium

Potassium, sodium, and calcium are also needed.

Literature Reviews

Gruenwald, J. (2014) "Human Outposts on Mars: Engineering and Scientific Lessons Learned from History."

• Identified basic issues in Mars colonization from analysis of concepts of historical human colonization.

• Hunger and malnutrition.

• Self Sufficiency

Khan et al. (2005) "Nutritional and therapeutic potential of Spirulina."

• Evaluated the nutritional and therapeutic value of Spirulina.

• Concluded that Spirulina can help treat variety of diseases.

Dao-lun Feng, and Zu-cheng Wu (2006) "Culture of Spirulina platensis in human urine for biomass production and O2 evolution."

● Cultivated Spirulina using real (RHU) and artificial (ZM) urine.

● Their urine was diluted 180-fold.

● Trace elements were added.

● Achieved similar growth to known media.

• Can Spirulina be cultivated effectively using a composite of nutrients derived from Mars regolith simulant and urine simulant?

Question

Hypothesis• If Mars regolith simulant is used in

unison with urine simulant, then it will have greater growth than if used separately.

Methods: Photobioreactor• A photobioreactor is

an apparatus for producing ample heat, light and, aeration/agitation suitable to spirulina growth.

Methods: Photobioreactor

Methods: Spirulina Inoculum • Spirulina inoculum was

acquired from Algae Research Supply

• Inoculum was stored on a shaker table running at 100 RPM until nutrient media was formulated.

Methods: ARS Medium

• Spirulina medium was also purchased from Algae Research Supply

• Was comprised of concentrated nutrient media and buffering salts that would be mixed with water

Methods: Urine Simulant• Urine simulant was

prepared as described by Shmaefsky et al 1992.

• Due to the high salt content, the urine was diluted by adding 3 parts distilled water to 1 part urine simulant.

Methods: Mars Regolith Simulant• Martian regolith

simulant (1 kg of JSC Mars-1A) was purchased

• The regolith simulant consisted of soils collected from a volcanic site in Hawaii.

Methods: Mars Dirt Solution• To extract any water soluble

nutrients and elements that may aid spirulina growth, Martian Regolith Simulant was added to 1 gallon of distilled water in a large flask.

• A magnetic stir table mixed the solution at 300 RPM for 3 days.

Methods: Mars Dirt Solution

• The solution was filtered through filter paper.

• A sample of MDS was sent off to Elemental Analysis Inc. to find out what elements made their way into the water.

Methods: pH Buffering• MDS and Urine

Simulant is slightly acidic with a pH of 6.5.

Methods: pH Buffering

• To raise and buffer the pH, sodium bicarbonate and sodium carbonate were added.

Methods: Culture Medium Dilution Layout

50% Urine +50% MDS

50% Urine +50% MDS

50% Urine +50% MDS

50% Urine +50%H20

50% Urine +50%H20

50% Urine +50%H20

ARS Spirulina Medium

ARS Spirulina Medium

ARS Spirulina Medium

50% MDS +50%H20

50% MDS +50%H20

50% MDS +50%H20

Methods: Measurement of Spirulina (Hemocytometer)• Only spirulina in the square of the

hemocytometer are counted.• Measured the number of turns in the

helix filament and the total amount of filaments .

Methods: Measurement of the Spirulina (Dry Weight) • After the 20 day

growing period, all cultures were filtered with filter paper.

• Following filtration, the papers were dried overnight.

Methods: Measurement of the Spirulina (Dry Weight) • Once dried, the papers

were weighed and averaged with corresponding samples.

• An unused piece of filter paper served as the weight comparison.

Results

Growth Average: Filaments

Growth Average: Turns

Visuals: Day 5

Visuals: Day 15

Visuals: Day 20

Dry WeightDry Weight Averages

Average Weight (Milligrams)

MDS 273.2

50% Urine, 50% MDS

9.8

Urine 222.4

ARS Spirulina Medium

170.3

Elemental Analysis of MDS

● All elements detected in MDS do aid the growth of spirulina.

● Due to detection limits in the tests, no nitrogen, phosphorus, or other trace elements were detected.

● More detailed tests are needed.

0.38%

48 ppm

2.8 ppm

24.8 ppm

Conclusions• All alternative mediums successfully allowed for the

cultivation of spirulina. • However, all cultures containing urine simulant lysed.

• We can infer that with further research, Martian astronauts can use the inexhaustible supply of urine and Martian regolith to grow spirulina and provide their food supply.

Discussion• Our results indicate that spirulina can be

cultivated using Martian resources.

• Is spirulina is truly the optimal method for feeding astronauts on our neighbor planet?

Future Work• Repeat testing and have more trials• Find/create a better Mars regolith simulant (Perchlorates) • Test different urine simulants or use real urine• Use different light levels• Use a different control medium• Different dilutions of urine to MDS• Investigate chemical reactions that may have caused cell lysis. • Compare with other food sources that could be used on Mars• Have a more detailed elemental analysis done• Earthbound Applications

ReferencesAllen, Carlton C., et al. "JSC Mars-1-Martian regolith simulant." Lunar and Planetary Science Conference. Vol. 28. 1997.

Allen, Carlton C., et al. "Martian soil simulant available for scientific, educational study." Eos, Transactions American Geophysical Union 79.34 (1998): 405-409.

Ciferri, Orio. "Spirulina, the edible microorganism." Microbiological reviews 47.4 (1983): 551.

Feng, Dao-lun, and Zu-cheng Wu. "Culture of Spirulina platensis in human urine for biomass production and O2 evolution." Journal of Zhejiang University SCIENCE B 7.1 (2006): 34-37.

Gruenwald, J. (2014) "Human Outposts on Mars: Engineering and Scientific Lessons Learned from History."CEAS Space J CEAS Space Journal 6.2 (2014): 73-77.

Khan, Zakir, Pratiksha Bhadouria, and P. S. Bisen. (2005) "Nutritional and therapeutic potential of Spirulina." Current pharmaceutical biotechnology 6.5 : 373-379.

Shmaefsky, Brian R. (1990) "Artificial Urine for Laboratory Testing." The American Biology Teacher 52.3: 170-72. Print.

Tuantet, Kanjana, Hardy Temmink, Grietje Zeeman, Marcel Janssen, René H. Wijffels, and Cees J.n. Buisman. (2014) "Nutrient Removal and Microalgal Biomass Production on Urine in a Short Light-path Photobioreactor." Water Research 55: 162-174.

Special Thanks To:• T.I.M.E. Science

• Mrs. J. Williams• Mr. M. Tuckey• Mrs. L. Patch• Dr. K. Wilcox

• T.C. 4-H• Mrs. M. Arnaudin

• Elemental Analysis Inc.• Orbitec• Pisgah Forest Rotary Club• Duke Energy• Transylvania County Schools• Viewers like you