Differential microbiota shift on whole romaine lettuce subjected to source or forward processing and on fresh-cut products during cold storage
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A research project in the food production group of the Closed Ecological Life Support System (CELSS) program sought to define optimum conditions for photosynthetic productivity of a higher plant food crop. The effects of radiation and various atmospheric compositions were studied.
Plants will be an important component of future long-term space missions. Lighting systems for growing plants will need to be lightweight, reliable, and durable, and light-emitting diodes (LEDs) have these characteristics. Previous studies demonstrated that the combination of red and blue light was an effective light source for several crops. Yet the appearance of plants under red and blue lighting is purplish gray making visual assessment of any problems difficult. The addition of green light would make the plant leave appear green and normal similar to a natural setting under white light and may also offer a psychological benefit to the crew. Green supplemental lighting could also offer benefits, since green light can better penetrate the plant canopy and potentially increase plant growth by increasing photosynthesis from the leaves in the lower canopy. In this study, four light sources were tested: 1) red and blue LEDs (RB), 2) red and blue LEDs with green fluorescent lamps (RGB), 3) green fluorescent lamps (GF), and 4) cool-white fluorescent lamps (CWF), that provided 0%, 24%, 86%, and 51% of the total PPF in the green region of the spectrum, respectively. The addition of 24% green light (500 to 600 nm) to red and blue LEDs (RGB treatment) enhanced plant growth. The RGB treatment plants produced more biomass than the plants grown under the cool-white fluorescent lamps (CWF treatment), a commonly tested light source used as a broad-spectrum control.
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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.
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Addition of chlorine-based antimicrobial substances to fresh-cut leafy green wash water is done to minimize microbial cross-contamination during processing. We developed the FDA Leafy Green Risk Assessment Model (FDA-LGRAM) to quantify the impact of free chlorine concentration in wash water during fresh-cut lettuce processing on the extent of water-mediated cross-contamination between shredded lettuce and the associated risk of illness due to exposure to Escherichia coli O157:H7. At different contamination prevalence and levels of E. coli O157:H7 on incoming lettuce heads, the model compared the predicted prevalence of contaminated fresh-cut lettuce packages and the risk of illness per serving between: (1) a scenario where fresh-cut lettuce was packaged without washing; and (2) scenarios involving washing fresh-cut lettuce with different levels of free chlorine (0 ppm, 5 ppm, 10 ppm, 15 ppm, and 20 ppm) prior to packaging. Our results indicate that the free chlorine level in wash water has a substantial impact on the predicted prevalence of contaminated fresh-cut lettuce packages and the risk of illness associated with E. coli O157:H7 in fresh-cut lettuce. Results showed that the required level of free chlorine that can minimize water-mediated cross-contamination and reduce the corresponding risk of illness depended on contamination prevalence and levels of E. coli O157:H7 on incoming lettuce heads. Our model also indicated that the pathogen inactivation rate in wash water via free chlorine was a key model parameter that had a significant impact on the extent of cross-contamination during washing and the predicted associated risk of illness.
The Salad Machine Research has continued to be a two path effort with the research at Marshall Space Flight Center (MSFC) focusing on the design, construction, and operation of a semiautomated system (Salad Machine) for the production of salad vegetables within a standard rack. Boeing Corporation in cooperation with NASA MSFC constructed a four drawer Salad Machine which was occasionally placed within the Space Station Freedom Mockup facility for view by selected visitors. Final outfitting of the Salad Machine is awaiting the arrival of parts for the nutrient delivery system. Research at the Alabama A&M facilities focused on compatibility of radish and lettuce plants when grown on the same nutrient solution. Lettuce fresh weight shoot yield was significantly enhanced when lettuce plants were grown on nutrient solution which was shared with radish. Radish tuber production was not significantly affected although there was a trend for radish from shared solutions to be heavier than those grown on separate nutrient solutions. The effect of sharing nutrient solutions on carbohydrate partitioning reflected the effect of sharing solution on fresh weight yield. Lettuce shoot dry weight was significantly greater for plants from shared solutions than from separate. There was no significant effect on sharing nutrient solution on radish tuber dry weight. Partitioning of nitrogen, calcium, magnesium, and potassium was not affected by sharing, there was, however, a disproportionate amount of potassium in the tissues, suggesting luxury consumption of potassium in all plants and tissues. It is concluded that lettuce plants benefit from sharing nutrient solution with radish and that radish is not harmed.
The ability of West Indian manatees (Trichechus manatus) to move between fresh and salt water raises the question of whether manatees drink salt water. Water turnover rates were estimated in captive West Indian manatees using the deuterium oxide dilution technique. Rates were quantified in animals using four experimental treatments: (1) held in fresh water and fed lettuce (N=4), (2) held in salt water and fed lettuce (N=2), (3) acutely exposed to salt water and fed lettuce (N=4), and (4) chronically exposed to salt water with limited access to fresh water and fed sea grass (N=5). Animals held in fresh water had the highest turnover rates (145+/-12 ml kg-1 day-1) (mean +/- s.e.m.). Animals acutely exposed to salt water decreased their turnover rate significantly when moved into salt water (from 124+/-15 to 65+/-15 ml kg-1 day-1) and subsequently increased their turnover rate upon re-entry to fresh water (146+/-19 ml kg-1 day-1). Manatees chronically exposed to salt water had significantly lower turnover rates (21+/-3 ml kg-1 day-1) compared with animals held in salt water and fed lettuce (45+/-3 ml kg-1 day-1). Manatees chronically exposed to salt water and fed sea grass had very low turnover rates compared with manatees held in salt water and fed lettuce, which is consistent with a lack of mariposia. Manatees in fresh water drank large volumes of water, which may make them susceptible to hyponatremia if access to a source of Na+ is not provided.
Several varieties of leafy vegetables were evaluated with the goal of selecting those with the best growth, nutrition, and organoleptic acceptability for ISS. Candidate species were narrowed to commercially available cultivars with desirable growth attributes for space (e.g., short stature and rapid growth). Seeds were germinated in controlled environment chambers under conditions similar to what might be found in the Veggie plant growth chamber on ISS. Eight varieties of leafy greens were grown: 'Tyee' spinach, 'Flamingo' spinach, 'Outredgeous' Red Romaine lettuce, 'Waldmann's Dark Green' leaf lettuce, 'Bull's Blood' beet, 'Rhubarb' Swiss chard, 'Tokyo Bekana' Chinese cabbage, and Mizuna. Plants were harvested at maturity and biometric data on plant height, diameter, chlorophyll content, and fresh mass were obtained. Tissue was ground and extractions were performed to determine the tissue elemental content of Potassium (K), Magnesium (Mg), Calcium (Ca) and Iron (Fe). Following the biometric/elemental evaluation, four of the eight varieties were tested further for levels of anthocyanins, antioxidant (ORAC-fluorescein) capacity, lutein, zeaxanthin, and Vitamin K. For sensory evaluation, 'Outredgeous' lettuce, Swiss chard, Chinese cabbage, and Mizuna plants were grown, harvested when mature, packaged under refrigerated conditions, and sent to the JSC Space Food Systems Laboratory. Tasters evaluated overall acceptability, appearance, color intensity, bitterness, flavor, texture, crispness and tenderness. All varieties received acceptable scores with overall ratings greater than 6 on a 9-point hedonic scale. Chinese cabbage was the highest rated, followed by Mizuna, 'Outredgeous' lettuce, and Swiss chard. Based on our results, the selected varieties of Chinese cabbage, lettuce, Swiss chard and Mizuna seem suitable for a pick-and-eat scenario on ISS with a ranking based on all factors analyzed to help establish priority.
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.
Plant growth in a controlled ecological life support system may entail the build-up over extended time periods of phytotoxic concentrations of volatile organic compounds produced by the plants themselves. Ethylene is a prominent gaseous emission of plants, and is the focus of this report. The objective was to determine the rate of ethylene release by spring wheat, white potato, and lettuce during early, middle, and late growth stages, and during both the light and dark segments of the diurnal cycle. Plants grown hydroponically using the nutrient film technique were covered with plexiglass containers for 4 to 6 h. At intervals after enclosure, gas samples were withdrawn with a syringe and analyzed for ethylene with a gas chromatograph. Lettuce produced 10 to 100 times more ethylene than wheat or potato, with production rates ranging from 141 to 158 ng g-dry/wt/h. Wheat produced from 1.7 to 14.3 ng g-dry/wt/h, with senescent wheat producing the least amount and flowering wheat the most. Potatoes produced the least amount of ethylene, with values never exceeding 5 ng g-dry/wt/h. Lettuce and potatoes each produced ethylene at similar rates whether in dark period or light period. Ethylene sequestering of 33 to 43 percent by the plexiglass enclosures indicated that these production estimates may be low by one-third to one-half. These results suggest that concern for ethylene build-up in a contained atmosphere should be greatest when growing lettuce, and less when growing wheat or potato.
Ethylene production by 20-sq m stands of wheat, soybean, lettuce and potato was monitored throughout growth and development in NASA's Controlled Ecological Life Support System (CELSS) Biomass Production Chamber. Chamber ethylene concentrations rose during periods of rapid growth for all four species, reaching 120 parts per billion (ppb) for wheat, 60 ppb for soybean, and 40 to 50 ppb for lettuce and potato. Following this, ethylene concentrations declined during seed fill and maturation (wheat and soybean), or remained relatively constant (potato). Lettuce plants were harvested during rapid growth and peak ethylene production. The highest ethylene production rates (unadjusted for chamber leakage) ranged from 0.04 to 0.06 ml/sq m/day during rapid growth of lettuce and wheat stands, or approximately 0.8 to 1.1 ml/g fresh weight/h. Results suggest that ethylene production by plants is a normal event coupled to periods of rapid metabolic activity, and that ethylene removal or control measures should be considered for growing crops in a tightly closed CELSS.
The 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. A systematic evaluation of rooting media and nutrient delivery systems were evaluated for three lettuce cultivars that have shown promise as candidates for a surface based food production system. 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 test. A Hazard Analysis and Critical Control Point (HACCP) evaluation was performed using ProSan(tm) as sanitation agent prior to consumption was approved, and the crew was allowed to consume the lettuce grown using the VEGGIE light cap and gravity based nutrient delivery system at the completion of the 14-day DRAT field test. The DRAT field test validated the crew operations; Growth of all lettuce cultivars was excellent. 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 excursion module. A number of areas have been identified for future work, to minimize the "footprint" of the Food Production system through creative use of unused wall and floor space in the unit.
The 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. A systematic evaluation of rooting media and nutrient delivery systems were evaluated for three lettuce cultivars that have shown promise as candidates for a surface based food production system. 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 test. A Hazard Analysis and Critical Control Point (HACCP) evaluation was performed using ProSantm as sanitation agent prior to consumption was approved, and the crew was allowed to consume the lettuce grown using the VEGGIE light cap and gravity based nutrient delivery system at the completion of the 14-day DRAT field test. The DRAT field test validated the crew operations; Growth of all lettuce cultivars was excellent. 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 excursion module. A number of areas have been identified for future work, to minimize the "footprint" of the Food Production system through creative use of unused wall and floor space in the unit.
Edible biomass from four crops of wheat (Triticum aestivum L.), four crops of lettuce (Lactuca sativa L.), four crops of potato (Solanum tuberosum L.), and three crops of soybean (Glycine max (L.) Merr.) grown in NASA's CELSS Biomass Production Chamber were analyzed for proximate composition. All plants were grown using recirculating nutrient (hydroponic) film culture with pH and electrical conductivity automatically controlled. Temperature and humidity were controlled to near optimal levels for each species and atmospheric carbon dioxide partial pressures were maintained near 100 Pa during the light cycles. Soybean seed contained the highest percentage of protein and fat, potato tubers and wheat seed contained the highest levels of carbohydrate, and lettuce leaves contained the highest level of ash. Analyses showed values close to data published for field-grown plants with several exceptions: In comparison with field-grown plants, wheat seed had higher protein levels; soybean seed had higher ash and crude fiber levels; and potato tubers and lettuce leaves had higher protein and ash levels. The higher ash and protein levels may have been a result of the continuous supply of nutrients (e.g., potassium and nitrogen) to the plants by the recirculating hydroponic culture.