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VEG-04: The Effects of Light Quality on Mizuna Mustard Growth, Nutritional Composition, and Organoleptic Acceptability for a Space Diet

Growing fresh, nutritious, palatable produce for crew consumption during spaceflight may provide health-promoting, bioavailable nutrients and enhance the astronaut dietary experience as we move toward longer-duration missions. Tending plants may also serve as a countermeasure for crew psychological stresses associated with spaceflight. However, requirements to support consistent growth of a variety of high quality, nutritious crops under spaceflight environmental conditions remain unclear. This study explores the potential to grow crops for consumption on the International Space Station (ISS) using the Veggie vegetable-production system. VEG-04A and B were two flight tests conducted in 2019 with the leafy green crop mizuna mustard. Mizuna was grown in two Veggie chambers simultaneously, with the chambers set to different red-to-blue light formulations; one Veggie was programmed as "red-rich" and the second as "blue-rich." Light quality is known to impact plant growth, nutrition, microbiology, and organoleptic characteristics on Earth, and the Veggie flight tests examined how these impacts might differ in microgravity. VEG-04A, a 35-day growth test with a single harvest, was initiated in June and harvested in July 2019. At harvest, the astronauts froze half of the edible plant tissue to return to Earth and weighed the remaining half using the Mass Measurement Device (MMD). Weighed samples were then cleaned with produce-sanitizing wipes, and consenting crew members participated in organoleptic evaluation of the fresh produce. The remaining sanitized produce was available for crew consumption as desired. Frozen flight samples were returned at the end of August for microbial and chemical analyses to assess food safety and nutritional quality. No pathogens were detected on VEG-04A flight or ground control samples. On average, bacterial and fungal counts were significantly lower on ground control samples than flight samples. VEG-04B, a 56-day test with multiple harvests from the same plants, assessed sustained productivity. VEG-04B was initiated in October 2019 with three harvests at four, six, and eight weeks after initiation. Challenges with the watering program occurred early during VEG-04A, and several plants failed to survive in both the flight and ground control operations. Thus, prior to VEG-04B, an extra test was conducted to tailor water timing and volumes. This test determined that mizuna grew best if the wicks inside the plant pillow were allowed to dry after plants germinated, reducing persistent water around the stem. The wicks changed from being a conduit for water out of the plant pillow to being a conduit for air into the root zone. This test allowed a fine tuning of methods for VEG-04B. It is our hope that these tests on ISS will help mitigate the risk of an inadequate food supply for long-duration missions by adding fresh vegetables to the crew diet. This research was co-funded by the Human Research Program and Space Biology (MTL#1075) in the ILSRA 2015 NRA call.

Massa, Gioia D.↗

Pick-and-Eat Space Crop Production Flight Testing on the International Space Station

Growing fresh, nutritious, palatable produce for crew consumption during spaceflight may provide health-promoting, bioavailable nutrients and enhance the astronaut dietary experience as we move toward longer-duration missions. However, requirements to support consistent growth of a variety of high-quality crops under spaceflight environmental conditions remain unclear. This study explores the potential to grow crops for consumption on the International Space Station (ISS) using Veggie, NASA’s vegetable production chamber system. VEG-04A and VEG-04B were two flight tests with ground components conducted with the leafy green crop mizuna mustard. In each location, mizuna was grown in two Veggie units simultaneously, with the chambers set to different red-to-blue-to-green light formulations. Preflight verification testing with various lighting treatments was conducted to down-select two treatments that contributed to the best desirable growth and sensory qualities in mizuna mustard. For the flight tests, one Veggie was programmed as “red-rich” with an average of 270 μmol m-2 s-1 of 630 nm red light, 30 μmol m-2 s-1 of 455 nm blue light, and 30 μmol m-2 s-1 of 530 nm green light. The second Veggie was “blue-rich” with an average of 150 µmol m-2 s-1 of 630 nm red light, 150 µmol m-2 s-1 of 455 nm blue light, and 30 µmol m-2 s-1 of 530 nm green light. Light quality is known to impact plant growth, nutrition, microbiology, and sensory characteristics on Earth, and the Veggie flight tests examined how these impacts might differ in microgravity. VEG-04A, a 35-day growth test with a single harvest, was initiated in June and harvested in July 2019. VEG-04B, a 56-day test with three harvests from the same plants, assessed sustained productivity. Preflight testing for VEG-04B was conducted after the VEG-04A flight test to improve the operations, approaches, and watering requirements, which resulted in better crop establishment in the VEG-04B flight test. VEG-04B was initiated in October 2019 with harvests at four, six, and eight weeks after initiation. At all of the harvests, the astronauts froze half of the edible plant tissue to return to Earth and weighed the remaining half using the Mass Measurement Device (MMD). Weighed samples were then cleaned with produce-sanitizing wipes, and consenting crew members participated in sensory evaluations of the fresh produce. The remaining sanitized produce was available for crew consumption as desired. Frozen flight samples were returned to Earth for chemical and microbial analyses to assess nutritional quality and food safety. Flight-grown mizuna was generally more acceptable to the crew and had higher nutrient levels, although mizuna grown in the ground control performed better in terms of higher yield and lower microbial load. This presentation will focus on results from the nutritional and sensory analyses, including how nutrients identified as key for supplementing the crew diet varied across lighting treatments, harvest approaches, and spaceflight versus ground conditions. It is our hope that these tests on the ISS will help mitigate the risk of an inadequate food supply for long-duration missions by adding fresh vegetables to the crew diet. This study was supported by NASA’s Human Research and Space Biology Programs through the HERO NNJ13ZSA002N-ILSRA grant solicitation.

Jess Bunchek↗

Veggie: Space Vegetables for the International Space Station and Beyond

The Veggie vegetable production system was launched to the International Space Station (ISS) in 2014. Veggie was designed by ORBITEC to be a compact, low mass, low power vegetable production system for astronaut crews. Veggie consists of a light cap containing red, blue, and green LEDs, an extensible transparent bellows, and a baseplate with a root mat reservoir. Seeds are planted in plant pillows, small growing bags that interface with the reservoir. The Veggie technology validation test, VEG-01, was initiated with the first test crop of 'Outredgeous' red romaine lettuce. Prior to flight, lettuce seeds were sanitized and planted in a substrate of arcillite (baked ceramic) mixed with controlled release fertilizer. Upon initiation, astronauts open the packaged plant pillows, install them in the Veggie hardware, and prime the system with water. Operations include plant thinning, watering, and photography. Plants were grown on the ISS for 33 days, harvested, and returned frozen to Earth for analysis. Ground controls were conducted at Kennedy Space Center in controlled environment chambers reproducing ISS conditions of temperature, relative humidity, and CO2. Returned plant samples were analyzed for microbial food safety and chemistry including elements, antioxidants, anthocyanins and phenolics. In addition the entire plant microbiome was sequenced, and returned plant pillows were analyzed via x-ray tomography. Food safety analyses allowed us to gain approvals for future consumption of lettuce by the flight surgeons and the payload safety office. A second crop of lettuce was grown in 2015, and the crew consumed half the produce, with the remainder frozen for later analysis. This growth test was followed by testing of a new crop in Veggie, zinnias. Zinnias were grown to test a longer duration flowering crop in preparation for tests of tomatoes and other fruiting crops in the future. Zinnias were harvested in February. Samples from the second harvest of lettuce and the zinnia harvest are frozen on the ISS and will return with the next cargo return flight. Some challenges occurred in all tests, especially in the area of watering, with plants receiving insufficient or excess water leading to stressed growth. Zinnia plants were also challenged with fungal growth. Initial tests with Veggie have given us great insight into future crop production scenarios as we work to develop regular supplemental salad crop production on ISS and larger food production systems for our journey to Mars. Funding for this research comes from NASA's Space Biology Program.

Food Safety↗

Space shuttle/food system. Volume 2, Appendix C: Food cooling techniques analysis. Appendix D: Package and stowage: Alternate concepts analysis

The relative penalties associated with various techniques for providing an onboard cold environment for storage of perishable food items, and for the development of packaging and vehicle stowage parameters were investigated in terms of the overall food system design analysis of space shuttle. The degrees of capability for maintaining both a 40 F to 45 F refrigerated temperature and a 0 F and 20 F frozen environment were assessed for the following cooling techniques: (1) phase change (heat sink) concept; (2) thermoelectric concept; (3) vapor cycle concept; and (4) expendable ammonia concept. The parameters considered in the analysis were weight, volume, and spacecraft power restrictions. Data were also produced for packaging and vehicle stowage parameters which are compatible with vehicle weight and volume specifications. Certain assumptions were made for food packaging sizes based on previously generated space shuttle menus. The results of the study are shown, along with the range of meal choices considered.

Source record↗

Modelling Groundwater Depletion at Regional and Global Scales: Present State and Future Prospects.

Except for frozen water in ice and glaciers, groundwater is the world's largest distributed store of freshwater and has strategic importance to global food and water security. In this paper, the most recent advances quantifying groundwater depletion (GWD) are comprehensively reviewed. This paper critically evaluates the recently advanced modeling approaches estimating GWD at regional and global scales, and the evidence of feedbacks to the Earth system including sea-level rise associated with GWD. Finally, critical challenges and opportunities in the use of groundwater are identified for the adaption to growing food demand and uncertain climate.

Groundwater Depletion (GWD)↗

Impacts of Groundwater Pumping on Regional and Global Water Resources

Except frozen water in ice and glaciers (68%), groundwater is the world's largest distributed store of freshwater (30%), and has strategic importance to global food and water security. In this chapter, the most recent advances assessing human impact on regional and global groundwater resources are reviewed. This chapter critically evaluates the recently advanced modeling approaches quantifying the effect of groundwater pumping in regional and global groundwater resources and the evidence of feedback to the Earth system including sea‐level rise associated with groundwater use. At last, critical challenges and opportunities are identified in the use of groundwater to adapt to growing food demand and uncertain climate.

groundwater↗

Rodent Research on the International Space Station - A Look Forward

Rodent Research on the International Space Station (ISS) is one of the highest priority science activities being supported by NASA and is planned for up to two flights per year. The first Rodent Research flight, Rodent Research-1 (RR-1) validates the hardware and basic science operations (dissections and tissue preservation). Subsequent flights will add new capabilities to support rodent research on the ISS. RR-1 will validate the following capabilities: animal husbandry for up to 30 days, video downlink to support animal health checks and scientific analysis, on-orbit dissections, sample preservation in RNA. Later and formalin, sample transfer from formalin to ethanol (hindlimbs), rapid cool-down and subsequent freezing at -80 of tissues and carcasses, sample return and recovery. RR-2, scheduled for SpX-6 (Winter 20142015) will add the following capabilities: animal husbandry for up to 60 days, RFID chip reader for individual animal identification, water refill and food replenishment, anesthesia and recovery, bone densitometry, blood collection (via cardiac puncture), blood separation via centrifugation, soft tissue fixation in formalin with transfer to ethanol, and delivery of injectable drugs that require frozen storage prior to use. Additional capabilities are also planned for future flights and these include but are not limited to male mice, live animal return, and the development of experiment unique equipment to support science requirements for principal investigators that are selected for flight. In addition to the hardware capabilities to support rodent research the Crew Office has implemented a training program in generic rodent skills for all USOS crew members during their pre-assignment training rotation. This class includes training in general animal handling, euthanasia, injections, and dissections. The dissection portion of this training focuses on the dissection of the spleen, liver, kidney with adrenals, brain, eyes, and hindlimbs. By achieving and maintaining proficiency in these basic skills as part of the nominal astronaut training curriculum this allows the rodent research program to focus the mission specific crew training on scientific requirements of research and operations flow.

Experimental capabilities↗

Drosophila Habitat Developed to Support Research on the International Space Station

The Fruit Fly Lab is a hardware suite being designed to support research on the International Space Station (ISS) for use by the entire Drosophila research community. A validation mission will launch and return on SpaceX-5 in late 2014, followed by Principal Investigator-lead science flights thereafter. Space flight experiments are selected via peer-reviewed proposals open to the Drosophila community. The cassettes (containers) that will house the Drosophila cultures were successfully used to conduct an immunity study on the Space Shuttle. Results showed that the innate immune system of Drosophila melanogaster was affected by space flight with a reduction in phagocytosis function of plasmatocytes, changes in antimicrobial peptides and other gene expression levels, as well as changes in development of the animals. Scientific research topics that are of interest to NASA will be presented. Each cassette used to house the Drosophila has a removable food tray that can be replaced to sustain the growth of the culture, or can be transferred to another cassette, along with embryos and burrowed larvae, enabling multi-generational studies. The cassette can be frozen in the Minus Eighty Laboratory Freezer for ISS to preserve samples until post-flight analysis, expanding the applications of the hardware. Utilization of a centrifuge allows for on-orbit 1g controls for microgravity experiments, as well as variable g-levels for lunar or Mars environment studies. The standard form factor used also allows for implementation of modular upgrades. An observation system, circadian rhythm lighting system, and fixation capability are upgrades currently in development for near-term implementation. This hardware suite, with its flight- proven design and ability to utilize existing on-board facilities, offers the whole Drosophila research community a platform to address several key areas of the National Research Councils decadal survey, supporting the utilization of ISS for science discovery.

Drosophila↗

Myco-Architecture off Planet: Growing Surface Structures at Destination

Our work focused on filling major select key technical knowledge gaps, and technical aspects to be addressed in a Phase II proposal were identified such as the secretion of glues or plastics by the fungi or bacteria to form biocomposites. Specifically, we raised the technical TRL by assessing the growth of in-house mycelial-producing strains on potential food substrates and analyzed the advantages and disadvantages to their use off planet. Growth of the mycelia on sawdust and powdered nutrients including dried, powdered algae, was used as a baseline. We measured growth as a function of temperature, tested the relevant material properties of the mycelia products produced, and measured the mass of input and output volume for production per volume of material. Mycotecture was produced in a bag simulating the proposed mission implementation. Post-production such as heat treatment (as is done with terrestrial mycotecture)was assessed.The material properties of the dried, frozen and baked mycelial outputs included density, strength,thermal insulation and flame retardation. Tensile/compression testing on biocomposite material specimens was performed to obtain three-dimensional elastic constants, as well as to examine deformation and fracture behavior. These mechanical tests were performed on both dormant and activated samples to understand the evolution of the structural material. In addition, experimental data obtained from the mechanical testing was used to build a failure prediction model that accounts for material anisotropy. This aided in future structural design through a quantitative understanding of the mechanical limits of the material. To examine the texture, defects, fracture surfaces of the material, high-resolution microstructural imaging was utilized before and after the mechanical tests. This microstructural analysis informed us about the macro-structural behavior and influenced structural design. Thermal decomposition analysis was also performed tounderstand the thermal limits.

Biocomposites↗

VEG-01: Veggie Hardware Verification Testing

The Veggie plant/vegetable production system is scheduled to fly on ISS at the end of2013. Since much of the technology associated with Veggie has not been previously tested in microgravity, a hardware validation flight was initiated. This test will allow data to be collected about Veggie hardware functionality on ISS, allow crew interactions to be vetted for future improvements, validate the ability of the hardware to grow and sustain plants, and collect data that will be helpful to future Veggie investigators as they develop their payloads. Additionally, food safety data on the lettuce plants grown will be collected to help support the development of a pathway for the crew to safely consume produce grown on orbit. Significant background research has been performed on the Veggie plant growth system, with early tests focusing on the development of the rooting pillow concept, and the selection of fertilizer, rooting medium and plant species. More recent testing has been conducted to integrate the pillow concept into the Veggie hardware and to ensure that adequate water is provided throughout the growth cycle. Seed sanitation protocols have been established for flight, and hardware sanitation between experiments has been studied. Methods for shipping and storage of rooting pillows and the development of crew procedures and crew training videos for plant activities on-orbit have been established. Science verification testing was conducted and lettuce plants were successfully grown in prototype Veggie hardware, microbial samples were taken, plant were harvested, frozen, stored and later analyzed for microbial growth, nutrients, and A TP levels. An additional verification test, prior to the final payload verification testing, is desired to demonstrate similar growth in the flight hardware and also to test a second set of pillows containing zinnia seeds. Issues with root mat water supply are being resolved, with final testing and flight scheduled for later in 2013.

Massa, Gioia↗

Probiotics in the Space Food System: Delivery, Microgravity Effects, and the Potential Benefit to Crew Health

As mission distance and duration increase, the need grows for non‐invasive disease prevention and immunomodulation, especially given the limited medical response capability expected for these missions and the immune dysregulation documented in crew. Additionally, changes in diet, lifestyle, antibiotic usage, and the environmental stresses during spaceflight may alter crewmembers' intestinal microbiome. The addition of probiotic bacteria to the space food system is expected to confer immunostimulatory benefits on crewmembers, with the potential to counteract the immune dysregulation that has been documented in spaceflight. Based on previous studies that demonstrated unique microbiological responses to the low shear environment of spaceflight, probiotic organisms hold the potential to induce enhanced beneficial responses through mechanisms, such as beneficial interactions with human immune cells and repression of colonization of pathogens on the mucosa. The work presented here will begin to address two research gaps related to providing probiotics in spaceflight: 1) delivery, and 2) the effect of the low shear microgravity environment on probiotic attributes. The probiotic Lactobacillus acidophilus was selected for investigation due to its wide commercial use and documented benefits that include inhibition of virulence related gene expression in pathogens and mucosal stimulation of immune cells. The delivery system for probiotics has not been determined for spaceflight, where the food system is shelf stable and the lack of refrigeration prevents the use of traditional dairy delivery methods. In order to demonstrate the potential of the space food system to deliver viable probiotic bacteria to crewmembers, the probiotic L. acidophilus was packaged in high barrier flight packaging in nonfat dry milk (NFDM) or retained in commercial capsule form. Viable cells were enumerated over 8 months of storage at 22, 4, and ‐80ºC. The survival of L. acidophilus rehydrated in NFDM, in a PBS control, and directly from the capsule was also evaluated following stress challenge with simulated gastric and intestinal juices to determine the method that would deliver the most viable cells to the intestine, where they would be expected to confer beneficial effects. L. acidophilus was found to be stable to gastric and intestinal juice challenge when delivered in rehydrated NFDM, even after two hours of exposure. In comparison, L. acidophilus was reduced by 1‐5 logs when exposed to gastric and intestinal juice directly and when rehydrated in a PBS control. Shelf life data indicated that L. acidophilus would require refrigerated or frozen storage to remain viable at adequate levels over the multi‐year storage periods required for spaceflight. This study suggests that the protective effect provided by the dairy matrix, and not merely rehydration prior to consumption, extends probiotic viability and stress tolerance during storage in spaceflight and in simulated digestion conditions more adequately than a capsule. In addition to effective delivery, it is essential to understand the microgravity effects on the stress tolerances and genetic expression of probiotic bacteria to enable optimization of growth, survival, strain selection, and conferred benefits in spaceflight. In our current study, ground‐based characterization of the growth, stress response, and transcriptomic response of L. acidophilus will be accomplished using the low shear modeled microgravity (LSMMG) culture environment in the rotating wall vessel (RWV). We hypothesize that L. acidophilus will maintain or improve its growth and stress response following culture in the LSMMG environment, and that transcriptomic analysis will define the associated molecular mechanism(s), resulting in the ability to optimize strain selection.

Castro, S. L.↗

Bionutrients-1, On-Demand Production of Nutrients in Space

Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small “production pack” system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.

Hindupur, Aditya↗

BioNutrients-1: On-Demand Production of Nutrients in Space

Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small "production pack" system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.

Hindupur, Aditya↗

Veggie ISS Validation Test Results and Produce Consumption

The Veggie vegetable production system flew to the International Space Station (ISS) in the spring of 2014. The first set of plants, Outredgeous red romaine lettuce, was grown, harvested, frozen, and returned to Earth in October. Ground control and flight plant tissue was sub-sectioned for microbial analysis, anthocyanin antioxidant phenolic analysis, and elemental analysis. Microbial analysis was also performed on samples swabbed on orbit from plants, Veggie bellows, and plant pillow surfaces, on water samples, and on samples of roots, media, and wick material from two returned plant pillows. Microbial levels of plants were comparable to ground controls, with some differences in community composition. The range in aerobic bacterial plate counts between individual plants was much greater in the ground controls than in flight plants. No pathogens were found. Anthocyanin concentrations were the same between ground and flight plants, while antioxidant and phenolic levels were slightly higher in flight plants. Elements varied, but key target elements for astronaut nutrition were similar between ground and flight plants. Aerobic plate counts of the flight plant pillow components were significantly higher than ground controls. Surface swab samples showed low microbial counts, with most below detection limits. Flight plant microbial levels were less than bacterial guidelines set for non-thermostabalized food and near or below those for fungi. These guidelines are not for fresh produce but are the closest approximate standards. Forward work includes the development of standards for space-grown produce. A produce consumption strategy for Veggie on ISS includes pre-flight assessments of all crops to down select candidates, wiping flight-grown plants with sanitizing food wipes, and regular Veggie hardware cleaning and microbial monitoring. Produce then could be consumed by astronauts, however some plant material would be reserved and returned for analysis. Implementation of this plan is a step toward developing pick-and-eat food production to supplement the packaged diet on ISS and for future exploration missions where plants could make up a larger portion of the diet. Supported by NASA Space Biology Program.

Food Safety↗

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗

Advanced Life Systems for Extreme Environments: An Arctic Application

The problems of obtaining adequate pure drinking water and disposing of liquid and solid waste in the U.S. Arctic, a region where virtually all water is frozen solid for much of the year, has led to unsanitary solutions (U.S. Arctic Research Commission). These solutions are also damaging to the environment. Sanitation and a safe water supply are particularly problems in rural villages. About one-fourth of Alaska's 86.000 Native residents live in these communities. They are without running water and use plastic buckets for toilets. The outbreak of diseases is believed to be partially attributable to exposure to human waste. Villages with the most frequent outbreaks of disease are those in which running water is difficult to obtain (Office of Technology Assessment, 1994). Waste is emptied into open lagoons, rivers, or onto the sea coast. It does not degrade rapidly and in addition to affecting human health, can be harmful to the fragile ecology of the Arctic and the indigenous wildlife and fish populations. Advanced Life Systems for Extreme Environments (ALSEE) provides a solution to sanitation and safe water problems. The system uses an advanced integrated technology developed for Antarctic and space applications. ALSEE uses the systems approach to address more than waste and water problems. By incorporating hydroponic horticulture and aquaculture into the waste treatment system, ALSEE addresses the quality and quantity of fresh foods available to Arctic residents. A temperate climate is required for year-round plant growth. ALSEE facilities can be designed to include a climate controlled area within the structure. This type of environment is a change from the long periods of darkness and cold found in the Arctic and can help alleviate stress so often associated with these extremes. While the overall concept of ALSEE projects is advanced, system facilities can be operated by village residents with appropriate training. ALSEE provides continuing training and education as a part of the project. Not only is this desirable but necessary. There is ample evidence in "gravoyard" throughout rural Alaska of technologies which failed not because they were not applic4tble, but because they were not integrated and could not be operated or repaired by village residents. Waste streams in villages will vary depending on the human diet and the non-food waste stream. ALSEE units can be adapted to different village situations once the content of the waste stream is known. Units located in population hubs can serve as research, education. and demonstration centers. Appropriate modifications can be determined at these centers to transfer technology to more remote locations where waste stream conditions may differ.

Lewis, Carol E.↗