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Identification of Fungal Colonies on Ground Control and Flight Veggie Plant Pillows

The Veggie system focuses on growing fresh produce that can be harvested and consumed by astronauts. The microbial colonies in each Veggie experiment are evaluated to determine the safety level of the produce and then differences between flight and ground samples. The identifications of the microbial species can detail risks or benefits to astronaut and plant health. Each Veggie ground or flight experiment includes six plants grown from seeds that are glued into wicks in Teflon pillows filled with clay arcillite and fertilizer. Fungal colonies were isolated from seed wicks, growth media, and lettuce (cv. 'Outredgeous') roots grown in VEG-01B pillows on ISS and in corresponding ground control pillows grown in controlled growth chambers. The colonies were sorted by morphology and identified using MicroSeq(TM) 500 16s rDNA Bacterial Identification System and BIOLOG GEN III MicroPlate(TM). Health risks for each fungal identification were then assessed using literature sources. The goal was to identify all the colonies isolated from flight and ground control VEG-01B plants, roots, and rooting medium and compare the resulting identifications.

Scotten, Jessica E.

Sustained Veggie: A Preliminary Look at Continuous On-Orbit Food Production

The Veggie system on the International Space Station (ISS) intermittently supplements the crew diet with fresh, leafy green crops. For 120 days, Sustained Veggie assessed the potential of continuous on-orbit crop production. Crops grown in Veggie have been grown concurrently, but Sustained Veggie staggered plant initiation and harvest to provide more constantly available produce. The objective of this preliminary study was to compare two growth schemes to determine the methodology for required inputs, optimal yield, food safety, and crew considerations.

Bunchek, Jess M.

Potential of a Plant Gas Exchange Mechanistic Model to Predict Plant Transpiration in Veggie on ISS

Plants are an essential part of long-duration space travel, as they enable food production and contribute to air revitalization through photosynthesis, and water recycling through transpiration. Understanding their growth mechanisms is essential to use them to sustain human life in space. In particular, gas exchange – e.g., CO2 absorption and water transpiration – are modified in microgravity because of the lack of buoyancy-driven convection, and in the long run, this could result in impaired plant growth. Water absorbed by the plants mainly depends on their size and on environmental conditions (air temperature, humidity and ventilation), but in microgravity watering plants is a delicate operation – too much water results in flooded roots and too little water leads in a few hours to wilted plants. This is regularly experienced in the Veggie system on ISS, which enables small-scale food production in microgravity since 2014. This presentation explores how a mechanistic model of plant gas exchange can help predict plant transpiration in Veggie and thus better predict daily watering. For each plant, inputs on canopy leaf area (acquired with daily photos), air temperature and relative humidity in the plant compartment, as well as airspeed at the top of its canopy enable accurate predictions of transpiration in microgravity. This brings a better understanding of water movement through the plant in microgravity in relation to ventilation and plant size and would result in easier management of plant watering in Veggie. Ultimately, this work could be applied to any space crop production in microgravity and be used for water management and yield predictions. This work was funded by NASA Space Biology through NASA postdoctoral program / USRA.

Lucie Poulet

Potential of a Plant Gas Exchange Mechanistic Model to Predict Plant Transpiration in Veggie on ISS

Plants are an essential part of long-duration space travel, as they enable food production and contribute to air revitalization through photosynthesis, and water recycling through transpiration. Understanding their growth mechanisms is essential to use them to sustain human life in space. In particular, gas exchange – e.g., CO2 absorption and water transpiration – are modified in microgravity because of the lack of buoyancy-driven convection, and in the long run, this could result in impaired plant growth. Water absorbed by the plants mainly depends on their size and on environmental conditions (air temperature, humidity and ventilation), but in microgravity watering plants is a delicate operation – too much water results in flooded roots and too little water leads in a few hours to wilted plants. This is regularly experienced in the Veggie system on ISS, which enables small-scale food production in microgravity since 2014. This presentation explores how a mechanistic model of plant gas exchange can help predict plant transpiration in Veggie and thus better predict daily watering. For each plant, inputs on canopy leaf area (acquired with daily photos), air temperature and relative humidity in the plant compartment, as well as airspeed at the top of its canopy enable accurate predictions of transpiration in microgravity. This brings a better understanding of water movement through the plant in microgravity in relation to ventilation and plant size and would result in easier management of plant watering in Veggie. Ultimately, this work could be applied to any space crop production in microgravity and be used for water management and yield predictions. This work was funded by NASA Space Biology through NASA postdoctoral program / USRA.

Lucie Poulet

Plant Growth Research for Food Production: Development and Testing of Expandable Tuber Growth Module

Controlled and reliable growth of a variety of vegetable crops is an important capability for manned deep space exploration systems for providing nutritional supplementation and psychological benefits to crew members. Because current systems have been limited to leafy vegetables that require minimal root space, a major goal for these systems is to increase their ability to grow new types of crops, including tuber plants and root vegetables that require a large root space. An expandable root zone module and housing was developed to integrate this capability into the Vegetable Production System (Veggie). The expandable module uses a waterproof, gas-permeable bag with a structure that allows for root space to increase vertically throughout the growth cycle to accommodate for expanding tuber growth, while minimizing the required media mass. Daikon radishes were chosen as an ideal tuber crop for their subterraneous tuber size and rapid growth cycle, and investigations were done to study expanding superabsorbent hydrogels as a potential growth media. These studies showed improved water retention, but restricted oxygen availability to roots with pure gel media. It was determined that these hydrogels could be integrated in lower proportions into standard soil to achieve media expansion and water retention desired. Using the constructed module prototype and ideal gel and soil media mixture, daikon radishes are being grown in the system to test the capability and success of the system through a full growth cycle.

food production

VEGGIE Pillow Testing: Microbial Analysis of Cut-and-Come-Again Species Testing

With NASA focused on researching and developing technology for deep space missions, the need for a reliable supplementary food source must also be considered. For the ISS, resupplying the food source is more practical and cost effect since the facility is in low Earth orbit. However, as NASA attempts to push the frontier in space, the costs and distance for resupply will surely increase. Plants would contribute to the proportion of food and reduce the dependency on food from Earth. In addition, plants would provide oxygen production, carbon dioxide removal, and psychological benefits. As a result, a vegetable production system, VEGGIE, was developed for NASA to produce salad crops with minimal resources and study the beneficial effects. The VEGGIE pillow is a single use bag for growing crops that is used with the VEGGIE hardware. The VEGGIE pillow was tested with four different species of plants with the cut-and-come-again harvest method to determine the greatest yield. Instead of harvesting the entire plant, the harvest consisted of cutting leaves to allow the plant to regrow leaves. The harvest methods included cutting the plants weekly, bi-weekly, and monthly. A fifth plant species, radishes, was also harvested and replanted. Microbial load analysis and an ANOVA significance test were utilized. The data suggest that the two Brassica plants have the greatest yields; however, the microbial load is also greatest for the two plants per gram of fresh weight. Furthermore, the results support the reuse of pillows for multiple harvests as shown by the replanted radishes.

Nguyen, Bao-Thang

Effects of Residual Water System Silver on Space Crop Microbiome and Nutrient Content

Ionic silver (Ag+) is being investigated as a residual biocide for use in spacecraft potable water systems on future crewed missions. In addition to providing clean water to the crew and other life support system functions, the potable water is used to irrigate space crop production units such as the Vegetable Production System (Veggie) and the Advanced Plant Habitat (APH). We have evaluated the impact of different concentrations of Ag+ biocide solutions in comparison to a control in both substrate (arcillite-based) and substrate-less (hydroponics-based) growth set ups. Here, we provide evidence that increasing the concentration of silver in the irrigation water impacts the root zone microbiome in both setups, with plant growth and elemental nutrient content also affected in the hydroponic set up. This suggests a need for a silver removal step to achieve acceptable silver levels in irrigation water before application to space crops in a substrate-less hydroponics system. This removal step is also recommended for a substrate-based system, although it is not as critical as in a hydroponics system.

Aubrie O’Rourke

Fall 2012 Graduate Engineering Internship Summary

In the fall of 2012, I participated in the National Aeronautics and Space Administration (NASA) Pathways Intern Employment Program at the Kennedy Space Center (KSC) in Florida. This was my second internship opportunity with NASA, a consecutive extension from a summer 2012 internship. During my four-month tenure, I gained valuable knowledge and extensive hands-on experience with payload design and testing as well as composite fabrication for repair design on future space vehicle structures. As a systems engineer, I supported the systems engineering and integration team with the testing of scientific payloads such as the Vegetable Production System (Veggie). Verification and validation (V&V) of the Veggie was carried out prior to qualification testing of the payload, which incorporated a lengthy process of confirming design requirements that were integrated through one or more validatjon methods: inspection, analysis, demonstration, and testing. Additionally, I provided assistance in verifying design requirements outlined in the V&V plan with the requirements outlined by the scientists in the Science Requirements Envelope Document (SRED). The purpose of the SRED was to define experiment requirements intended for the payload to meet and carry out.

Ehrlich, Joshua

3D Printed Substrate

In preparation for long-term, manned, deep space missions, NASA requires a sustainable system for crop and food production. This system has a variety of benefits, including a fresh food supply, improvements in air quality, a lower need to resupply and psychological benefits for the gardeners. Specifically, NASA is attempting to improve and iterate their Vegetable Production System (VEGGIE), a plant growth unit currently aboard the International Space Station (ISS). The VEGGIE unit is highly dependent on a variety of factors, including passive water delivery, growth lights, rooting pillows and cabin conditions. While previous teams have improved and redesigned water reservoirs, the primary objective of the project was to create a new substrate unit to replace the current rooting pillow design. The current design consists of an electrostatic bag filled with arcillite and a slow-release fertilizer pellet. The prototype design retains the fertilizer pellet for nutrient consistency, however the outer bag and arcillite fillings have been replaced with a 3D printed lattice block. This lattice block has several key points that allow it to function in similar ways to the current design: 1. The lattice planes are porous, with each pore offset such that the overall porosity is the same as the arcillite filling. 2. The block demonstrates a wicking nature and is able to pull water upwards towards the roots. This minimizes time needed to integrate with the existing water reservoir. 3. The filament used is highly flexible and is able to pull apart to accommodate root growth. 4. The lattice planes are connected by microfibers left behind from the printing process. These fibers provide support for the growing roots and keep the lattice planes properly aligned. The substrate block design seeks to reduce the payload costs by being entirely 3D printed. While filament would still need to be provided to the ISS, it is far less expensive than shipping arcillite due to the significantly lower weight. The final design has iterated the lattice plane concept and utilizes vertically placed planes with pores running parallel to the water reservoir. This design has reliably shown water uptake and retention and has been successful in growing multiple romaine lettuce plants. Future work should include further growth testing using multiple plant species, compost and reusability testing, food safety testing and microgravity growth testing.

Affan Bhutta

Concept for Sustained Plant Production on ISS Using VEGGIE Capillary Mat Rooting System

Plant growth in microgravity presents unique challenges associated with maintaining appropriate conditions for seed germination, seedling establishment, maturation and harvest. They include maintaining appropriate soil moisture content, nutrient balance, atmospheric mixing and containment. Sustained production imposes additional challenges of harvesting, replanting, and safety. The VEGGIE is a deployable (collapsible) plant growth chamber developed as part of a NASA SBIR Phase II by Orbitec, Madison, WI. The intent of VEGGIE is to provide a low-resource system to produce fresh vegetables for the crew on long duration missions. The VEGGIE uses and LED array for lighting, an expandable bellows for containment, and a capillary matting system for nutrient and water delivery. The project evaluated a number of approaches to achieve sustained production, and repeated plantings, using the capillary rooting system. A number of different root media, seed containment, and nutrient delivery systems were evaluated and effects on seed germination and growth were evaluated. A number of issues limiting sustained production, such as accumulation of nutrients, uniform water, elevated vapor pressure deficit, and media containment were identified. A concept using pre-planted rooting packs shown to effectively address a number of those issues and is a promising approach for future development as a planting system for microgravity conditions.

Stutte, Gary W.

Plant Growth Research for Food Production: Development and Testing of Expandable Tuber Growth Module

Controlled and reliable growth of a variety of vegetable crops is an important capability for manned deep space exploration systems for providing nutritional supplementation and psychological benefits to crew members. Because current systems have been limited to leafy vegetables that require minimal root space, a major goal for these systems is to increase their ability to grow new types of crops, including tuber plants and root vegetables that require a large root space. An expandable root zone module and housing was developed to integrate this capability into the Veggie growth system. The expandable module uses a waterproof, gas-permeable bag with a structure that allows for root space to increase vertically throughout the growth cycle to accommodate for expanding tuber growth, while minimizing the required media mass. Daikon radishes were chosen as an ideal tuber crop for their subterraneous tuber size and rapid growth cycle, and investigations were done to study expanding superabsorbent hydrogels as a potential growth media. These studies showed improved water retention, but restricted oxygen availability to roots with pure gel media. It was determined that these hydrogels could be integrated in lower proportions into standard soil to achieve media expansion and water retention desired. Using the constructed module prototype and ideal gel and soil media mixture, Daikon radishes were grown in the system to test the capability and success of the system through a full growth cycle.

space

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

Growth of Three Lettuce Cultivars in NASA's HDU PEM During the 2010 DRATS Test

NASA's 2010 Desert Research and Technology Studies (DRATS) of the VEGGIE Food Production System in the Habitat Demonstration Unit (HDU) Pressurized Excursion Module (PEM) was the first operational evaluation of salad crop production technology in a NASA analog test. Rooting media and slow release fertilizers were evaluated for three lettuce cultivars that had shown promise as candidates for a surface based food production system. These tests involved comparing growth, color and quality of the lettuce cultivars grown under VEGGIE LED array (Orbitec, Madison, WI) or Biomass Production System for Education ((BSEe), Orbitec, Madison, WI) compact fluorescent lamps using a gravity feed water delivery system. Mission relevant conditions of CO2, temperature and RH were maintained using controlled environment chambers (EGC, Chagrin Falls, OH). Growth data was obtained for the two red leaf lettuce cultivars, Outredgeous and Firecracker, and the green Bibb lettuce cultivar, Flandria. Growth and quality was evaluated using different concentrations (7.5 g/L and 15 g/L) of commercial slow release fertilizer (Osmocote Plus 15-9-12, Scotts, Maryville, OH) and Nutricote 18-6-8 (Florikan, Sarasota, FL) in either a peat/vermiculite media (sunshine LP5 MiX, Sungro, Bellview, WA) or calcined montmorillonite clay [(arcillite,)Turface Proleague, Profile LLC, Buffalo Grove, IL]. The commercial peat/vermiculite mix generally resulted in larger plants than those grown in arcillite. Increasing the concentration of Osmocote from 7.5 to 15 g/L increased the height, dry mass, and leaf area of lettuce cultivars. In contrast, there was a decrease in growth parameters when concentration of Nutricote was increased from 7.5 to 15 g/L. The best growth was obtained with the 7.5 g/L Nutricote using a commercial peat/vermiculite mixture. This media was used for field testing VEGGIE plant system in the 2010 DRAT test. The VEGGIE nutrient delivery system worked well, was able to be maintained by multiple operators with a minimum of training, and supported excellent lettuce growth for the duration of the 14-day test. The operational DRAT field testing in the HDU identified light quality issues related to morphology and pigment development that will need to be addressed through additional testing. Feedback from the crew, ground support personnel, and human factors leads was uniformly positive on the psychological value of having the crop production system in the pressurized excursion module. Data are being used to design a plant atrium with LED lighting to evaluate salad crop growth during NASA's 2011 DRATS test.

Stutte, Gary W.

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.

Veggie

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.

LED light

Continued Environmental Microbiology Monitoring of The International Space Station (ISS) Veggie Unit Used for In-Flight, Crop-Based Food Systems

The International Space Station is a closed environment where rotating sets of Crewmembers live and work. This environment is monitored to ensure occupants’ health and safety during their spaceflight residency by routine Environmental Health System (EHS) collection of microbial samples including air, surface, and water. The microbial samples are collected, enumerated, and analyzed quarterly to monitor on-board system contamination and potential risks to crew health. Quarterly monitoring of the microorganisms in the ISS environment supports crew safety and contributes to a large set of microbial concentration and diversity data. The current in-inflight microbial requirements were developed using this historical data collected by the routine environmental monitoring. These in-flight microbial requirements have been established to maintain the health and safety of the spacecraft environment. This study leverages quarterly operational EHS sampling by collecting additional microbial samples from the surface of the Veggie plant production system on ISS. These samples will yield microbial concentration and diversity that can be compared and analyzed with nominal surface samples from the vehicle. The data collected in this study will aid in the development of requirements for spaceflight-based food production systems. Continued surface sampling of the internal and external surfaces of the Veggie system, along with collaboration from both Johnson Space Center (JSC) & Kennedy Space Center (KSC) scientists studying the microbiome of the veggie-crop systems, will be implemented as part of the future development of crop-based food system requirements for the ISS and beyond.

Christian Mena