Crop Readiness Level (CRL): A Scale to Track Progression of Crop Testing for Space

The development of engineering technologies and hardware for aerospace applications is often tracked on a 1-9 scale of readiness or TRL, with a "1" representing very basic or fundamental principles, and a "9" being flight tested, functional hardware. Preparing to grow crops for supplemental food and eventual life support contributions on space missions faces similar challenges. Nearly 20 years ago, the concept of a "crop readiness level" was suggested at a bioregenerative life support conference held at Kennedy Space Center, but there was little follow up to this. We propose to revive this concept to track the preparation and testing of different crop species for eventual use in the unique environment of space. For the sake of uniformity, we recommend a 1-9 scale, with a "1" being just the identification of a potential crop, followed by some basic horticultural testing, cultivars trials, then testing growth and yield under various controlled environments, progression to more space-like environments and hardware, understanding the nutritional, organoleptic, and food safety aspects of the crop, initial testing in space, and a final stage of growing the crop for food in space ("9"). We attempted to make the scaling logical and progressive, but our main goal is to initiate a dialogue in the space, plant research community to develop a scale for assessing crop readiness

New Frontiers in Food Production Beyond LEO

New technologies will be needed as mankind moves towards exploration of cislunar space, the Moon and Mars. Although many advances in our understanding of the effects of spaceflight on plant growth have been achieved in the last 40 years, spaceflight plant growth systems have been primarily designed to support space biology studies. Recently, the need for a sustainable and robust food system for future missions beyond Low Earth Orbit (LEO) has identified gaps in current technologies for food production. The goal is to develop safe and sustainable food production systems with reduced resupply mass and crew time compared to current systems

Does Seed Sanitization Affect the Plant Rhizosphere Microbiome and Its Ability to Compete with the Human Associated Pathogen, E. coli on Salad Crops?

Cultivation of crops in controlled environmental agricultural systems may limit microbial colonization and reduce diversity of the microbial communities. Practices like seed and growth medium sanitization may further impact microbial communities in the mature plant and the plants capacity to limit the growth of pathogens through competition. As humans expand their travels to space, understanding plant growth, health, and development in closed environments will be critical to the success of producing a safe, supplemental food source for astronauts. To determine the persistence of a potential human pathogen in plant growth and development, sanitized and unsanitized seeds from, mizuna (Brassica rapa var japonica) and red romaine lettuce (Lactuca sativa cultivar Outredgeous), were inoculated with Escherichia coli, ATCC 21445, germinated under simulated International Space Station (ISS) environmental conditions and harvested every 7 days until maturity. The persistence of E. coli in the rhizosphere was determined by plating on selective media, real time PCR (Polymerase Chain Reaction) and community sequencing of the rhizosphere communities. E. coli was detected in the crops roots and leaves for several weeks post germination. At day 28, plants from sanitized seeds had significantly higher counts of E. coli on the roots than those from unsanitized seeds. E. coli was also detected on a few uninoculated plants indicating airborne cross contamination among plants in the same growth chamber and suggesting an influence of the natural microbiome on human pathogen survival and persistence in leafy greens. Sequencing analysis revealed variations in composition and diversity between the communities. Understanding the microbial community of the rhizospheric microbiome is only the first step in determining the relationships between plants. Additional studies to include genotypic and phenotypic variations in the plants should be considered to determine if the natural microbes in the rhizosphere may contribute to the health and therefore, safety of the edible plants

Preparation for Pick-and-Eat Food Production on the International Space Station: Flight Definition for the VEG-04 and VEG-05 Missions

Growth of fresh, nutritious, palatable produce for crew consumption during spaceflight may provide health-promoting, bioavailable nutrients and enhance the dietary experience as we move toward longer-duration missions. Tending plants also may serve as a countermeasure for crew psychological stresses associated with long duration spaceflight. However, requirements to support consistent growth of a variety of high quality, nutritious crops under spaceflight environmental conditions is unknown. This study is exploring the potential to grow plants for food production on the International Space Station (ISS) using the Veggie vegetable production system. Ground testing is underway to compare the impacts of several fertilizer and lighting treatments on growth, quality, and nutritional composition of the leafy green crop mizuna, and the dwarf tomato crop "Red Robin" when subjected to Veggie ISS environmental conditions. Early testing focused on the leafy crop "Tokyo Bekana" Chinese cabbage, but ground tests indicated that this plant suffered from stress responses when grown under LEDs and the chronically elevated CO2 levels found on the ISS. Mizuna, a related leafy variety that grows well in the presence of high CO2, and has excellent organoleptic characteristics, was selected as an alternate crop. Tomato crops have been grown using two fertilizer formulations and two pollination techniques, and growth tests using different red:blue lighting environments are underway. Chemical analysis is also being conducted and these data, when coupled with the growth results, will be used to down-select to the two best lighting treatments and best fertilizer treatment for future testing of each crop on the ISS. Additionally, seed-source testing has become important, with mizuna seeds from two different vendors growing very differently. A seed source has been selected, and seed-surface-sanitizing methods have been confirmed for mizuna, but these remain under development for tomato. A crop-handling protocol is also being evaluated to support food safety. All harvests reserve a subset of samples for microbial analysis to determine baseline microbial levels and help establish critical control points for food safety. Testing was initially conducted in hardware analogs of the standard Veggie plant pillows. However, a new Veggie watering system, the Passive Orbital Nutrient Delivery System or PONDS, has been designed and is being prepared for future flight experiments. With the selection of this growth system, ground tests have shifted to analog PONDS systems. Crop tests on ISS, designated VEG-04 for mizuna and VEG-05 for tomato, are planned in 2018 to evaluate any additional impacts of spaceflight on the light and fertilizer conditions down-selected from ground tests. A set of Veggie-specific questions has been developed to characterize the psychological impacts of plant growth and plant-care activities during spaceflight. Organoleptic questionnaires have been developed to assess produce attributes in microgravity taste sessions. These tests for plants growing in the Veggie hardware on ISS will help to mitigate the risk of an inadequate food supply for long duration missions by developing methods and determining hardware requirements to integrate fresh vegetables as a dietary supplement. This research was co-funded by the Human Research Program and Space Biology (MTL#1075) in the ILSRA 2015 NRA call

Leafy Greens Grown on the International Space Station May Provide a Nutritious Supplement to Astronauts' Diet

Supplemental safe food production has been an essential goal of NASA to meet the nutritional needs of astronauts on the International Space Station (ISS) as well as for future long duration missions to the moon and beyond. Food crops grown in space experience different environmental conditions than plants grown on Earth (i.e. microgravity and spaceflight physical sciences impacts). To test the growth methods and effects of the space environment, red romaine lettuce Lactuca sativa cv. 'Outredgeous', was grown in Veggie plant growth chambers on the ISS. Microbiological food safety of the plants grown on the ISS was determined by heterotrophic plate counts to assess total microbial load for bacteria and fungi as well as screening for specific pathogens and isolate identification. Molecular characterization was completed using Next Generation Sequencing (NGS) to provide valuable information on the taxonomic composition and community structure of the plant microbiome. Chemical analyses of plant tissue were conducted to understand spaceflight-induced changes in key elements in the space diet, phenolics, anthocyanin levels, and Oxygen radical absorbance capacity (ORAC), a measure of antioxidant capacity. Three growth tests of red romaine lettuce were completed on ISS, VEG-01A, VEG-01B, and VEG-03A. Plants were harvested using two harvest methods, either a single terminal harvest (after 33 days) or cut-and-come-again repetitive harvesting (64 days total growth). Ground controls were grown simultaneously with a delay to accommodate condition monitoring and replication. A comparison of the plant tissue returned to Earth showed leaves from the second grow-out had significantly higher bacterial counts than the preceding or subsequent growth test or any of the ground controls. Fungal counts were significantly higher on the final cut-and-come-again harvest of the third grow out. None of the potential foodborne pathogens that were screened for were detected. Bacterial and fungal isolate identification and community characterization indicated similar diversity between VEG-01A and VEG-01B growth tests, however, there appeared to be subtle differences in diversity and distribution among the three growth tests. Chemical analysis of plant tissue revealed significant variation in a few elemental data, but variation in levels of phenolics, anthocyanins, and ORAC was not significantly different. This study indicated that leafy vegetable crops could safely provide an edible supplement to astronauts' diet, and our analysis provided baseline data for continual operation of the Veggie plant growth units on ISS. This research was funded by NASA's space biology program

Preparing for Veg-04 and Veg-05: Improving Pick-And-Eat Food Capabilities for the International Space Station

The capability to grow nutritious, palatable food for crew consumption during spaceflight has the potential to provide health-promoting, bioavailable nutrients, enhance the dietary experience, and reduce launch mass as we move toward longer-duration missions. Studies of edible produce during spaceflight have been limited, leaving a significant knowledge gap in the methods required to grow safe, acceptable, nutritious crops for consumption in space. Researchers from Kennedy Space Center, Johnson Space Center, Purdue University and ORBITEC have teamed up to explore the potential for plant growth and food production on the International Space Station (ISS) and future exploration missions. Ground testing of Chinese cabbage and dwarf tomato crops under different LED lighting and fertilizer conditions is being conducted to allow for a preliminary down selection of the two best lighting recipes and the best fertilizer treatment. Two trials of Chinese cabbage and one trial on dwarf tomato have been completed in on-going ground tests. Horticultural data on crop growth and productivity and chemical data on specific nutrients have been collected and are being analyzed to allow preliminary down selection. Taste test evaluations are planned on the preliminary down selection treatments to allow a final down selection for flight testing. Microbial assessment for hazard analysis critical control points (HACCP) evaluation is also underway to enable implementation of food consumption. Following down selection flight preparation will commence for testing these crops in the Veggie vegetable-production system on the ISS. A crew questionnaire has been developed to better understand the impact of crop growth in Veggie on crew behavioral health. A single Veggie plant growth chamber is currently installed on ISS, and preparations are underway to launch a second Veggie, allowing side-by-side testing under different lighting conditions. Veg-04 will be the first mission that will use this dual-Veggie capability, where the selected cultivar of Tokyo bekana Chinese cabbage will be grown under two different red-to-blue light ratios. ORBITEC has developed custom lighting software allowing independent selection of red and blue light levels. The VEG-05 experiment will test similar light treatments using Red Robin dwarf tomato. These tests offer an opportunity to develop a pick-and-eat fresh vegetable component to the ISS food system as a first step to regular supplemental food production. Our work will help define light colors, levels, and horticultural best practices to achieve high yields of safe, nutritious leafy greens and tomatoes to supplement a space diet of prepackaged food. With this work we will continue the synergistic research to help close gaps in the human research roadmap, and enable humans to venture to Mars and beyond. This research was co-funded by the Human Research Program and Space Biology (MTL1075) in the ILSRA 2015 NRA call

Preparation for Pick-and-Eat Food Production on the International Space Station: Flight Definition for the VEG-04 and VEG-05 Missions

Growth of fresh, nutritious, palatable produce for crew consumption during spaceflight may provide health-promoting, bioavailable nutrients and enhance the dietary experience as we move toward longer-duration missions. Tending plants also may serve as a countermeasure for crew psychological stresses associated with long duration spaceflight. However, requirements to support consistent growth of a variety of high quality, nutritious crops under spaceflight environmental conditions is unknown. This study is exploring the potential to grow plants for food production on the International Space Station (ISS) using the Veggie vegetable production system. Ground testing is underway to compare the impacts of several fertilizer and lighting treatments on growth, quality, and nutritional composition of the leafy green crop mizuna, and the dwarf tomato crop Red Robin when subjected to Veggie ISS environmental conditions. Early testing focused on the leafy crop Tokyo Bekana Chinese cabbage, but ground tests indicated that this plant suffered from stress responses when grown under LEDs and the chronically elevated CO2 levels found on the ISS. Mizuna, a related leafy variety that grows well in the presence of high CO2, and has excellent organoleptic characteristics, was selected as an alternate crop. Tomato crops have been grown using two fertilizer formulations and two pollination techniques, and growth tests using different red:blue lighting environments are underway. Chemical analysis is also being conducted and these data, when coupled with the growth results, will be used to down-select to the two best lighting treatments and best fertilizer treatment for future testing of each crop on the ISS. Additionally, seed-source testing has become important, with mizuna seeds from two different vendors growing very differently. A seed source has been selected, and seed-surface-sanitizing methods have been confirmed for mizuna, but these remain under development for tomato. A crop-handling protocol is also being evaluated to support food safety. All harvests reserve a subset of samples for microbial analysis to determine baseline microbial levels and help establish critical control points for food safety. Testing was initially conducted in hardware analogs of the standard Veggie plant pillows. However, a new Veggie watering system, the Passive Orbital Nutrient Delivery System or PONDS, has been designed and is being prepared for future flight experiments. With the selection of this growth system, ground tests have shifted to analog PONDS systems. Crop tests on ISS, designated VEG-04 for mizuna and VEG-05 for tomato, are planned in 2018 to evaluate any additional impacts of spaceflight on the light and fertilizer conditions down-selected from ground tests. A set of Veggie-specific questions has been developed to characterize the psychological impacts of plant growth and plant-care activities during spaceflight. Organoleptic questionnaires have been developed to assess produce attributes in microgravity taste sessions. These tests for plants growing in the Veggie hardware on ISS will help to mitigate the risk of an inadequate food supply for long duration missions by developing methods and determining hardware requirements to integrate fresh vegetables as a dietary supplement. This research was co-funded by the Human Research Program and Space Biology (MTL1075) in the ILSRA 2015 NRA call

Survival of E. Coli in the Rhizosphere and Phyllosphere of Leafy Greens Grown in Controlled Environment Chambers Under International Space Station Conditions

NASA's mission for manned long- duration space exploration drives the research for crop selection to provide a nutritious and safe supplement to an astronaut's diet. Understanding plant growth, health, and the associated microbial communities in closed environments will be critical to the success of this mission. Cultivation of crops in closed controlled environment agricultural systems may limit microbial colonization and reduce diversity of the microbial communities. Furthermore, practices like seed and growth medium sanitization may impact microbial communities in the mature plant and the capacity to limit the growth of food borne pathogens through competition

Rehabilitation of Multiple Ligament Knee Injuries

Work published in Clinical Orthopaedic Rehabilitation: An Evidence-Based Approach