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NASA NTRS2016

Risk of Visual Impairment and Intracranial Hypertension After Space Flight: Evaluation of the Role of Polymorphism of Enzymes Involved in One-Carbon Metabolism

Data from the Nutritional Status Assessment protocol provided biochemical evidence that the one-carbon metabolic pathway may be altered in individuals experiencing vision-related issues during and after space flight (1, 2). Briefly, serum concentrations of homocysteine, cystathionine, 2-methylcitric acid, and methylmalonic acid were significantly (P<0.001) higher (25-45%) in astronauts with ophthalmic changes than in those without such changes (1). These differences existed before, during, and after flight. Serum folate was lower (P<0.01) during flight in individuals with ophthalmic changes. Preflight serum concentrations of cystathionine and 2-methylcitric acid, and mean in-flight serum folate, were significantly (P<0.05) correlated with postflight changes in refraction (1). A follow-up study was conducted to evaluate a small number of known polymorphisms of enzymes in the one-carbon pathway, and to evaluate how these relate to vision and other medical aspects of the eye. Specifically, we investigated 5 polymorphisms in MTRR, MTHFR, SHMT, and CBS genes and their association with ophthalmic changes after flight in 49 astronauts. The number of G alleles of MTRR 66 and C alleles of SHMT1 1420 both contributed to the odds of visual disturbances (3). Block regression showed that B-vitamin status at landing and genetics were significant predictors for many of the ophthalmic outcomes studied (3). In conclusion, we document an association between MTRR 66 and SHMT1 1420 polymorphisms and space flightinduced vision changes. These data document that individuals with an altered 1-carbon metabolic pathway may be predisposed to anatomic and/or physiologic changes that render them susceptible to ophthalmic damage during space flight.

Smith, S. M.↗

NASA NTRS2015

Benefits of Microalgae for Human Space Exploration

Algae have long been known to offer a number of benefits to support long duration human space exploration. Algae contain proteins, essential amino acids, vitamins, and lipids needed for human consumption, and can be produced using waste streams, while consuming carbon dioxide, and producing oxygen. In comparison with higher plants, algae have higher growth rates, fewer environmental requirements, produce far less "waste" tissue, and are resistant to digestion and/or biodegradation. As an additional benefit, algae produce many components (fatty acids, H2, etc.) which are useful as biofuels. On Earth, micro-algae survive in many harsh environments including low humidity, extremes in temperature, pH, and as well as high salinity and solar radiation. Algae have been shown to survive inmicro-gravity, and can adapt to high and low light intensity while retaining their ability to perform nitrogen fixation and photosynthesis. Studies have demonstrated that some algae are resistant to the space radiation environment, including solar ultraviolet radiation. It remains to be experimentally demonstrated, however, that an algal-based system could fulfil the requirements for a space-based Bioregenerative Life Support System (BLSS) under comparable spaceflight power, mass, and environmental constraints. Two specific challenges facing algae cultivation in space are that (i) conventional growth platforms require large masses of water, which in turn require a large amount of propulsion fuel, and (ii) most nutrient delivery mechanisms (predominantly bubbling) are dependent on gravity. To address these challenges, we have constructed a low water biofilm based bioreactor whose operation is enabled by capillary forces. Preliminary characterization of this Surface Adhering BioReactor (SABR) suggests that it can serve as a platform for cultivating algae in space which requires about 10 times less mass than conventional reactors without sacrificing growth rate. Further work is necessary to compare the performance of microalgae-based systems, including SABR, with systems based on higher plants, as well as conventional physicochemical-based systems. Ongoing and future work in our laboratory is therefore directed determining the feasibility of using algae as a component of a BLSS in space.

Algae↗

NASA NTRS2016

Pick and Eat Crop Testing: Dwarf Tomato and Pepper as Candidate Space Crops

Several dwarf tomato and pepper varieties were evaluated under International Space Station (ISS)-simulated growth conditions (22 degrees Centigrade, 50 percent relative humidity, 1500 parts per million CO2, and 300 micromoles per square meter per second of light for 16 hours per day) with the goal of selecting those with the best growth, nutrition, and organoleptic potential for use in a pick and eat salad crop system on ISS and future exploration flights. Testing included six cultivars of tomato (Red Robin, Scarlet Sweet 'N' Neat, Tiny Tim, Mohamed, Patio Princess, and Tumbler) and six cultivars of pepper (Red Skin, Fruit Basket, Cajun Belle, Chablis, Sweet Pickle, and Pompeii). Plants were grown to an age sufficient to produce fruit (up to 106 days for tomato and 109 days for pepper) using Turface (arcillite) potting media with 18-6-8 control-release fertilizer and supplemental nutrient solution beginning around 60-days-age. Tomato fruits were harvested when they showed full red color, beginning around 70-days age and then at weekly intervals thereafter, while peppers were grown until fruits showed color and were harvested twice (first test) and just once at the end of the second test, with the final harvests including colored and green fruit. Plant sizes, yields, and nutritional attributes were measured and used to down-select to three cultivars for each species. In particular, we were interested in cultivars that were short (dwarf) but still produced high yields. Nutritional data included elemental (Ca, Mg, Fe, and K) content, vitamin K, phenolics, lycopene (for tomato), anthocyanin, lutein, and zeaxanthin. The three down-selected cultivars for each species were grown again and the harvested fruit sent to NASA's Johnson Space Center for sensory evaluation, which included overall acceptability, appearance, color intensity, aroma, flavor and texture. The combined data were compared and given weighting factors to rank the cultivars as candidates for testing in space. Weightings gave maximum importance to plant size (smaller being good) and fruit yield (greater yields being good). For tomato, the ranking was 1) cultivar Mohamed and cultivar Red Robin (tied), and 3) cultivar Sweet N' Neat. For pepper, the ranking was 1) cultivar Pompeii, 2) cultivar Red Skin, and 3) cultivar Fruit Basket. These rankings are somewhat subjective but provide a starting point for conducting higher fidelity testing with these crops (e.g., testing with light emitting diode lighting similar to the Veggie plant unit on ISS), and ultimately conducting a flight experiment.

Food↗

NASA NTRS2016

Food Fortification Stability Study

This study aims to assess the stability of vitamin content, sensory acceptability and color variation in fortified spaceflight foods over a period of 2 years. Findings will identify optimal formulation, processing, and storage conditions to maintain stability and acceptability of commercially available fortification nutrients. Changes in food quality are being monitored to indicate whether fortification affects quality over time (compared to the unfortified control), thus indicating their potential for use on long-duration missions.

Sirmons, T. A.↗

NASA NTRS2015

Nanoscale Test Strips for Multiplexed Blood Analysis

A critical component of the DNA Medicine Institute's Reusable Handheld Electrolyte and Lab Technology for Humans (rHEALTH) sensor are nanoscale test strips, or nanostrips, that enable multiplexed blood analysis. Nanostrips are conceptually similar to the standard urinalysis test strip, but the strips are shrunk down a billionfold to the microscale. Each nanostrip can have several sensor pads that fluoresce in response to different targets in a sample. The strips carry identification tags that permit differentiation of a specific panel from hundreds of other nanostrip panels during a single measurement session. In Phase I of the project, the company fabricated, tested, and demonstrated functional parathyroid hormone and vitamin D nanostrips for bone metabolism, and thrombin aptamer and immunoglobulin G antibody nanostrips. In Phase II, numerous nanostrips were developed to address key space flight-based medical needs: assessment of bone metabolism, immune response, cardiac status, liver metabolism, and lipid profiles. This unique approach holds genuine promise for space-based portable biodiagnostics and for point-of-care (POC) health monitoring and diagnostics here on Earth.

Chan, Eugene↗

NASA NTRS2016

Space Plant Biology Research at KSC

Long duration space exploration will require the capability for crews to grow their own food. Growing food is desirable from a mass-efficiency standpoint, as it is currently not feasible to carry enough prepackaged food on spacecraft to sustain crews for long duration missions. Nutritionally, fresh produce provides key nutrients that are not preserved well in pre-packaged meals (e.g. vitamins C and K) and those that are able to counteract detrimental effects of space flight, such as antioxidants to combat radiation exposure and lutein for decreasing macular degeneration. Additionally, there are significant psychological benefits of maintaining gardens, one being an indicator for the passage of time.

Veggie↗

NASA NTRS2017

Food Fortification Stability Study

This study aimed to assess the stability of vitamin content, sensory acceptability and color variation in fortified spaceflight foods over a period of two years. Findings will help to identify optimal formulation, processing, and storage conditions to maintain stability and acceptability of commercially available fortification nutrients. Changes in food quality were monitored to indicate whether fortification affects quality over time (compared to the unfortified control), thus indicating their potential for use on long-duration missions.

Sirmons, T. A.↗

NASA NTRS2018

Meal Replacement Mass Reduction Integration and Acceptability Study

The Orion Multi-Purpose Crew Vehicle (MPCV) and future exploration missions are mass constrained; therefore we are challenged to reduce the mass of the food system by 10% while maintaining safety, nutrition, and acceptability to support crew health and performance for exploration missions. Meal replacement with nutritionally balanced, 700-900 calorie bars was identified as a method to reduce mass. However, commercially available products do not meet the requirements for a meal replacement in the spaceflight food system. The purpose of this task was to develop a variety of nutritionally balanced, high quality, breakfast replacement bars, which enable a 10% food mass savings. To date, six nutrient-dense meal replacement bars have been developed, all of which meet spaceflight nutritional, microbiological, sensory, and shelf-life requirements. The four highest scoring bars were evaluated based on final product sensory acceptability, nutritional stability, qualitative stability of analytical measurements (i.e. color and texture), and microbiological compliance over a period of two years to predict long-term acceptability. All bars maintained overall acceptability throughout the first year of storage, despite minor changes in color and texture. However, added vitamins C, B1, and B9 degraded rapidly in fortified samples of Banana Nut bars, indicating the need for additional development. In addition to shelf-life testing, four bar varieties were evaluated in the Human Exploration Research Analog (HERA), campaign 3, to assess the frequency with which actual meal replacement options may be implemented, based on impact to satiety and psychosocial measurements. Crewmembers (n=16) were asked to consume meal replacement bars every day for the first fifteen days of the mission and every three days for the second half of the mission. Daily surveys assessed the crew's responses to bar acceptability, mood, food fatigue and perceived stress. Preliminary results indicate that the majority of crew members were noncompliant with daily meal replacement during the first half of the mission. Several crew members chose to forgo the meal, resulting in caloric deficits that were higher on skipped-bar days. Body mass loss was significant throughout the mission. Although there was no significant difference in body mass loss overall between the first half and second half of the mission, a higher number of individual crew members lost more body mass in the first half of the mission. Analysis is still ongoing, but current trends suggest that daily involuntary meal replacement can lead to greater individual impacts on body mass and psychological factors, while meal replacement on a more limited basis may be acceptable to most crew for missions up to 30 days. This data should be considered in Orion mass trades with health and human performance.

Sirmons, T.↗

NASA NTRS2017

Development of Storage Methods for Saccharomyces Strains to be Utilized for In situ Nutrient Production in Long-Duration Space Missions

Long-duration space missions will benefit from closed-loop life support technologies that minimize mass, volume, and power as well as decrease reliance on Earth-based resupply. A system for In situ production of essential vitamins and nutrients can address the documented problem of degradation of stored food and supplements. Research has shown that the edible yeast Saccharomyces cerevisiae can be used as an on-demand system for the production of various compounds that are beneficial to human health. A critical objective in the development of this approach for long-duration space missions is the effective storage of the selected microorganisms. This research investigates the effects of different storage methods on survival rates of the non-sporulating probiotic S. boulardii, and S. cerevisiae spores and vegetative cells. Dehydration has been shown to increase long-term yeast viability, which also allows increased shelf-life and reduction in mass and volume. The process of dehydration causes detrimental effects on vegetative cells, including oxidative damage and membrane disruption. To maximize cell viability, various dehydration methods are tested here, including lyophilization (freeze-drying), air drying, and dehydration by vacuum. As a potential solution to damage caused by lyophilization, the efficacy of various cryoprotectants was tested. Furthermore, in an attempt to maintain higher survival rates, the effect of temperature during long-term storage was investigated. Data show spores of the wild-type strain to be more resilient to dehydration-related stressors than vegetative cells of either strain, and maintain high viability rates even after one year at room temperature. In the event that engineering the organism to produce targeted nutrient compounds interferes with effective sporulation of S. cerevisiae, a more robust method for improving vegetative cell storage is being sought. Therefore, anhydrobiotic engineering of S. cerevisiae and S. boulardii is being conducted

nutrient↗

NASA NTRS2018

Hydroponics for Food Production in Space: History and Current Efforts

Goals of Food Production in Space: NASA Human Research Program (HRP) near-term food production systems - supplement key nutrients (Vitamins B1,C, K and potassium) that degrade to inadequate levels on exploration class missions; Study behavioral health aspect - caring for plants and improving the acceptability of the astronaut diet by supplementing it with fresh produce (currently anecdotal); Develop sustainable water delivery technologies - current TRL (Technological Readiness Level) 9 particulate based systems (i.e. used in research conducted in plant growth facilities APH (Advanced Plant Habitat), Veggie, BPS (Biomass Production System), LADA/SVET (Russian space plant chamber systems)) require resupply of bulky consumables and generate waste media.

Dreschel, T.↗

NASA NTRS2017

Development of Storage Methods for Saccharomyces Strains to be Utilized for In Situ Nutrient Production in Long-Duration Space Missions

life support↗

NASA NTRS2019

Dwarf Tomato and Pepper Cultivars for Space Crops

Several dwarf tomato and pepper varieties were evaluated under ISS-simulated growth conditions (22ºC, 50% RH, 1500 ppm CO2, and 300 μmol m(exp -2) s(exp -1) of light for 16 h per day) with the goal of selecting those with the best growth, nutrition, and organoleptic potential for use in a pick and eat salad crop system on ISS and future exploration flights. Testing included six cultivars of tomato (Red Robin, Scarlet Sweet ’N’ Neat, Tiny Tim, Mohamed, Patio Princess, and Tumbler) and six cultivars of pepper (Red Skin, Fruit Basket, Cajun Belle, Chablis, Sweet Pickle, and Pompeii). Plants were grown to an age sufficient to produce fruit (70 to 106 days for tomato and 109 days for pepper). Tomato fruits were harvested when they showed full red color, beginning ca. 70-days age and then at weekly intervals thereafter, while peppers were grown until numerous fruits showed color and all fruits (green and colored) were harvested once at the end of the test. Plant sizes, yields, and nutritional attributes were measured and used to down-select to three cultivars for each species. In particular, we were interested in cultivars that were short (dwarf) but still produced high yields. Nutritional data included elemental (Ca, Mg, Fe, and K) composition, vitamin K, phenolics, lycopene, anthocyanin, lutein, and zeaxanthin. The three down-selected cultivars for each species were evaluated for sensory attributes, including overall acceptability, appearance, color intensity aroma, flavor and texture. The combined data were compared and given weighting factors to rank the cultivars as potential candidates for testing in space. For tomato, the ranking was 1) cv. Mohamed, 2) cv. Red Robin, and 3) cv. Sweet N’ Neat. For pepper, the ranking was 1) cv. Pompeii, 2) cv. Red Skin, and 3) cv. Fruit Basket. These rankings are somewhat subjective but provide a good starting point for conducting higher fidelity testing with these crops (e.g., testing with LED lighting similar to the Veggie plant unit), and ultimately conducting flight experiments.

Spencer, LaShelle E.↗

NASA NTRS2019

New Technologies for Enabling Food Production Beyond LEO

NASA has identified the need for robust and sustainable ‘Pick-and-Eat’ systems for supplementing crew diets with fresh leafy green crops in near-term LEO (Low Earth Orbit), cislunar, and lunar missions. Spaceflight plant growth systems have been primarily designed for conducting space biology studies, but these systems are not optimal for sustained food production. Improved water and nutrient delivery subsystems that do not use bulky and non-reusable media are needed for decreasing the mass of the food production system. Autonomous technologies for monitoring plant health and food safety are needed for ensuring that the food produced is suitable supplementing crew diets with fresh, nutritious salad crops. Improved plant imaging techniques used for high-throughput phenotyping can be leveraged for monitoring plant health. Near-real-time measurements of the microbial ecology of food production systems are needed for assessing food safety. Furthermore, newly identified plant species and cultivars with improved growth habits and contents of antioxidants, vitamins, and minerals when grown in spaceflight environmental conditions are needed. These improvements in food production technologies will enable the design of sustainable life support systems for manned exploration missions beyond Low Earth Orbit.

Monje Mejia, Oscar A.↗

NASA NTRS2019

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↗

NASA NTRS2019

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↗

NASA NTRSDate not supplied

Changes in Plants Developed from Imbibed Seeds Irradiated by Simulated Solar Particle Events

One of the major concerns for long-term exploration missions beyond the Earth’s magnetosphere is radiation risk primarily from solar particle events (SPEs) and galactic cosmic rays (GCRs). With the goal of enabling manned Mars exploration, the production of fresh food during long duration space missions may provide nutritional supplementation and benefit astronauts’ behavioral health. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. In this study, we evaluated the effect of simulated SPEs on imbibed seeds of mizuna mustard, ‘Outredgeous’ red romaine lettuce, and ‘Red Robin’ dwarf tomato. Seeds were exposed to various doses of simulated SPEs (0, 40, 80 cGy, and 200 cGy), either acutely or at a low dose rate (LDR only for 40 and 80 cGy doses), using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Simulated SPE exposure was achieved using sequential exposures to protons with a broad energy range at 50-150 MeV/n. Control and irradiated seeds were planted either onto 0.5% MS-based agar plates containing required nutrients for early seedling analysis, or in peat plugs/pots for 28 day (mizuna and lettuce) and 91 day (tomato) grow outs. All plants were maintained under space-vehicle relevant conditions at 3000 ppm CO2, 50% RH, and 23°C, and a 16/8 h photoperiod with 300 μmol·m(exp -2)·s(exp -1) photosynthetically active radiation. Plant/fruit morphometrics and edible fresh mass were measured. Additionally, nutrient analyses were performed on mizuna and tomato fruit. Seedlings from irradiated seeds (both 40 and 80 cGy) showed significant and dose dependent impacts on cotyledon development and root length. Interestingly, small, but significant changes in vitamin content were found in mizuna and tomato fruits grown from 80 cGy (LDR) irradiated seeds. In addition, 80 cGy (LDR) simulated SPEs significantly delayed tomato early seedling development and plant growth, and reduced the quantity of fruits produced from the surviving plants. (This study was supported by NASA Human Research Program.)

J. T. Richards↗

NASA NTRS2020

Lunar Lettuce Production During Artemis III Mission to the Moon's South Pole

The Artemis Science Plan identified that the infrastructure and resources associated during a human/robotic exploration mission can be used to conduct fundamental lunar science needed to mitigate the risks of human exploration of surface systems (i.e. Mars and the Moon). In particular, studying the response of life to the combination of fractional gravity and deep space radiation of the Moon is needed to mitigate the HEOMD risks to the crew that result from inadequate diets. Essential nutrients and vitamins available to astronauts have been shown to degrade within the stored food system during long duration missions. Other risks to exploration missions were isolation and confinement, distance from the Earth, radiation and partial gravity.

O. Monje↗

NASA NTRSDate not supplied

Changes in Plants Developed from Imbibed Seeds Irradiated by Simulated Solar Particle Events

One of the major concerns for long-term exploration missions beyond the Earth’s magnetosphere is radiation risk primarily from solar particle events (SPEs) and galactic cosmic rays (GCRs). With the goal of manned Mars exploration, the production of fresh food during long duration space missions may provide nutritional supplementation and benefit astronauts’ behavioral health. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. In this study, we evaluated the effect of simulated SPEs on imbibed seeds of mizuna mustard, ‘Outredgeous’ red romaine lettuce, and ‘Red Robin’ dwarf tomato. Seeds were exposed to various doses of simulated SPEs (0, 40, 80 cGy, and 200 cGy), either acutely or at a low dose rate (LDR only for 40 and 80 cGy doses), using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Simulated SPE exposure was achieved using sequential exposures to protons with a broad energy range at 50-150 MeV/n. Control and irradiated seeds were planted either onto 0.5% MS-based agar plates containing required nutrients for early seedling analysis, or in peat plugs/pots for 28 day (mizuna and lettuce) and 91 day (tomato) grow outs. All plants were maintained under space-vehicle relevant controlled environment conditions at 3000 ppm CO2, 50% RH, and 23°C, and a 16/8 h photoperiod with 300 μmol·m-2·s-1 photosynthetically active radiation. Plant/fruit morphometrics and edible fresh mass were measured. In addition, nutrient analyses were performed for both mizuna and ripe tomatoe fruit. Seedlings from irradiated seeds (both 40 cGy and 80 cGy) showed significant changes in cotyledon development, as well as root length. Interestingly, small, but significant changes in vitamin content were found in mizuna and tomato fruits grown from 80 cGy (LDR) irradiated seeds. In addition, 80 cGy (LDR) simulated SPEs significantly affected tomato early seedling development, plant growth, and the quantity of fruits produced from the surviving plants. Generally, compared to 80 cGy, 40 cGy (LDR) of simulated SPEs displayed some impacts to the seeds and seedlings, but to a much lesser degree. (This study was supported by NASA Human Research Program.)

J.T. Richards↗