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Hummerick, Mary E.

Publications and source records attributed to Hummerick, Mary E..

At least 19 records

How Does Water Delivery System Design Impact the Microbial Load of Salad Crops?

In a microgravity setting, such as the environment aboard the International Space Station (ISS), an ideal plant water delivery system is one that can grow edible crops with minimal resource consumption and minimal risk to crew members. There are also concerns associated with the ability to control fluid escape and biofilm formation resulting in potential dangers to systems, crops, or crewmembers. To identify an appropriate system, candidate systems were assembled and operated under simulated ISS environmental conditions (T,CO2,and RH) with red romaine lettuce (Lactuca sativa cultivar 'Outredgeous') as a model crop. Fluid reservoirs and randomly selected planting sites were sampled every seven days until maturity at which point edible plant biomass and root samples were also taken. Heterotrophic bacteria and fungi growth patterns throughout each planting cycle were determined by plate counts on appropriate agar media. The candidate systems were compared to a classic hydroponics system as a control and harvested crops were compared to controls as well as Veggie-grown and market produce. Plants harvested from candidate systems yielded lower average heterotrophic bacteria and fungi per gram of plant mass levels when compared to market and Veggie samples as well as those from the control system. Additional studies to evaluate the system sanitation regimen as well as testing additional crops should be considered to aid in the selection of an ideal system.

Curry, Aaron B.

Plant Microbiomes May Provide Vital Information to Plant Success

Plant associated microbiomes, the rhizosphere and phyllosphere, are composed of communities of bacteria and fungi that may be mutualistic or pathogenic. These communities have the potential to influence plant health and development and can affect plant growth. Crop plants are being investigated as a fresh and safe supplement to astronauts’ diet and it is critical to understand and characterize these microbial communities. Multi-species crops, Mizuna mustard (Brassica rapa var japonica), ‘Outredgeous’ red romaine lettuce (Lactuca sativa), and Waldman’s Green lettuce (Lactuca sativa) were grown in two Veggie units on the International Space Station (ISS) for three grow outs in various combinations of plant types. Upon harvest, plant and pillow samples were frozen and returned to Earth for analysis. Bacterial and fungal community analyses for plant leaf and root, as well as pillow components, wick and media, were completed using next generation sequencing with the goal of surveying the composition of the entire community and identifying any potential pathogens. Bacteria were identified using the 16S rRNA gene whereas, fungi were identified with the internal transcribed spacer (ITS). The community composition for these three crops was compared between crop types and between plant tissue types. It is vital to mission success for the short term and long term to add nutritious, safe to eat vegetables providing a supplement to the crew members’ dietary requirements as well as to develop planning for deep space missions as we reach for the moon and on to Mars. Veggie technology validation tests were supported by NASA’s Space Biology Program.

Khodadad, Christina L.

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 plant’s 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.

Khodadad, Christina L. M.

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.

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.

Hummerick, Mary E.

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.

Food Production

Investigating Microbiome Differences Between Red Romaine Lettuce Grown from Sanitized and Unsanitized Seeds

The International Space Station (ISS) as an integral component for the discovery and development of advanced robotics, materials, communications, medicine, agriculture, and environmental science due to it currently being the world's only microgravity laboratory of its kind. Because the ISS is a contained system with confined quarters, much research has been undertaken to assess and diminish the number of microbiological risks associated with astronauts inhabiting the station for extended periods of time. Notable microbiological risk factors include drinking water, air, and food. As an avenue for both mental/emotional respite and a source of fresh produce for astronauts, a vegetable production system has been employed on the ISS. In order to understand the microbial risks involved with a "pick and eat" vegetable system on the International Space Station (ISS), this study aims to compare microbial differences between sanitized and unsanitized seeds by tracking and identifying seedborne microbes throughout the development of red romaine lettuce (Lactuca sativa)-a plant species that has already been grown on the ISS.

Polanco, Jonilee A.

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.

Evaluation of Low-Pressure Cold Plasma for Disinfection for ISS Grown Produce and Metallic Instrumentation

Cold plasma (CP) cleaning is a dry, non-thermal process, which can provide broad-spectrum antimicrobial activity yet reportedly causes little to no damage to the object being sanitized. Since cold plasma uses no liquids, it has the distinct advantage when used in microgravity of not having to separate liquids from the item being cleaned. This paper will present results on an effort to use low pressure CP to disinfect or sterilize materials for in space applications. Exposure times from 0 to 60 minutes and pressures ranging from 10 to 100 Pa were used to optimize plasma parameters to achieve acceptable kill rates for 3 bacteria, Bacillus cereus, E. coli and Bacillus pumulis SAFR-32 and one fungi, Aspergillus niger. These tests were done on produce and metal coupons to simulate medical equipment. Produce testing was not successful, with unacceptable kill rates and the produce being negatively impacted by exposure to the plasma. The plasma caused a 5 log reduction in the number of viable bacteria on metal coupon tests, which placed the number of viable bacteria below the detection limit. This is a very promising result showing that sterilization of medical equipment with cold plasma is feasible. Scanning Electron Microscope images were taken before and after exposure. The images after plasma exposure shows that the bacteria spores have been physically affected, as their size has gotten smaller and the appearance has changed.

Hintze, Paul E.

Evaluation of Low-Pressure Cold Plasma for Disinfection of ISS Grown Produce and Metallic Instrumentation

Cold plasma (CP) cleaning is a dry, non-thermal process, which can provide broad-spectrum antimicrobial activity yet reportedly causes little to no damage to the object being sanitized. Since cold plasma uses no liquids, it has the distinct advantage when used in microgravity of not having to separate liquids from the item being cleaned. This paper will present results on an effort to use low pressure CP to disinfect or sterilize materials for in space applications. Exposure times from 0 to 60 minutes and pressures ranging from 0.10 to 1.0 mbar were used to optimize plasma parameters. Tests were done on produce and metal coupons to simulate medical equipment. Escherichia coli was used as the challenge organism on produce and Bacillus pumilus SAFR-32 was used on metal surfaces. Produce testing was not successful, with unacceptable kill rates and the produce being negatively impacted by exposure to the plasma. The plasma caused a 5 log reduction in the number of viable bacteria on metal coupon tests, which placed the number of viable bacteria below the detection limit. This is a very promising result showing that sterilization of medical equipment with cold plasma is feasible. Scanning Electron Microscope images were taken before and after exposure. The images after plasma exposure show that the bacteria spores have been physically affected, as their size has gotten smaller and their appearance has changed.

Hintze, Paul E.

Hollow Fiber Membrane Bioreactor Systems for Wastewater Processing: Effects of Environmental Stresses Including Dormancy Cycling and Antibiotic Dosing

Membrane-aerated biofilm reactors (MABRs) have been studied for a number of years as an alternate approach for treating wastewater streams during space exploration. While the technology provides a promising pre-treatment for lowering organic carbon and nitrogen content without the need for harsh stabilization chemicals, several challenges must be addressed before adoption of the technology in future missions. One challenge is the transportation of bioreactors containing intact, active biofilms as a means for rapid start-up on the International Space Station or beyond. Similarly, there could be a need for placing these biological systems into a dormant state for extended periods when the system is not in use, along with the ability for rapid restart. Previous studies indicated that there was little influence of storage condition (4 or 25 C, with or without bulk fluid) on recovery of bioreactors with immature biofilms (48 days old), but that an extensive recovery time was required (20+ days). Bioreactors with fully established biofilms (13 months) were able to recover from a 7-month dormancy within 4 days (approximately 1 residence). Further dormancy and recovery testing is presented here that examines the role of biofilm age on recovery requirements, repeated dormancy cycle capabilities, and effects of long-duration dormancy cycles (8-9 months) on HFMB systems. Another challenge that must be addressed is the possibility of antibiotics entering the wastewater stream. Currently, for most laboratory tests of biological water processors, donors providing urine may not contribute to the study when taking antibiotics because the effects on the system are yet uncharacterized. A simulated urinary tract infection event, where an opportunistic, pathogenic organism, E. coli, was introduced to the HFMBs followed by dosing with an antibiotic, ciprofloxacin, was completed to study the effect of the antibiotic on reactor performance and to also examine the development of antibiotic-resistant communities within the system.

Bioreactor Systems

Inflight Microbial Monitoring- An Alternative Method to Culture Based Detection Currently Used on the International Space Station

Previous research has shown that potentially destructive microorganisms and human pathogens have been detected on the International Space Station (ISS). The likelihood of introducing new microorganisms occurs with every exchange of crew or addition of equipment or supplies. Microorganisms introduced to the ISS are readily transferred between crew and subsystems (i.e. ECLSS, environmental control and life support systems). Current microbial characterization methods require enrichment of microorganisms and at least a 48-hour incubation time. This increases the microbial load while detecting only a limited number of the total microorganisms. The culture based method detects approximately 1-10% of the total organisms present and provides no identification. To identify and enumerate ISS microbes requires that samples be returned to Earth for complete analysis. Therefore, a more expedient, low-cost, in-flight method of microbial detection, identification, and enumeration is warranted. The RAZOR EX, a ruggedized, commercial off the shelf, real-time PCR field instrument was tested for its ability to detect microorganisms at low concentrations within one hour. Escherichia coli, Salmonella enterica Typhimurium, and Pseudomonas aeruginosa were detected at low levels using real-time DNA amplification. Total heterotrophic counts could also be detected using a 16S gene marker that can identify up to 98% of all bacteria. To reflect viable cells found in the samples, RNA was also detectable using a modified, single-step reverse transcription reaction.

Inflight Microbial

Inflight Microbial Monitoring - An Alternative Method to Culture Based Detection Currently Used on the International Space Station

Microorganisms including potential human pathogens have been detected on the International Space Station (ISS). The potential to introduce new microorganisms occurs with every exchange of crew or addition of equipment or supplies. Current microbial monitoring methods require enrichment of microorganisms and a 48-hour incubation time resulting in an increase in microbial load, detecting a limited number of unidentified microorganisms. An expedient, low-cost, in-flight method of microbial detection, identification, and enumeration is warranted.

International Space Stateion

Dormancy and Recovery Testing for Biological Wastewater Processors

Bioreactors, such as aerated membrane type bioreactors have been proposed and studied for a number of years as an alternate approach for treating wastewater streams for space exploration. Several challenges remain before these types of bioreactors can be used in space settings, including transporting the bioreactors with their microbial communities to space, whether that be the International Space Station or beyond, or procedures for safing the systems and placing them into dormant state for later start-up. Little information is available on such operations as it is not common practice for terrestrial systems. This study explored several dormancy processes for established bioreactors to determine optimal storage and recovery conditions. Procedures focused on complete isolation of the microbial communities from an operational standpoint and observing the effects of: 1) storage temperature, and 2) storage with or without the reactor bulk fluid. The first consideration was tested from a microbial integrity and power consumption standpoint; both room temperature (25 C) and cold (4 C) storage conditions were studied. The second consideration was explored; again, for microbial integrity as well as plausible real-world scenarios of how terrestrially established bioreactors would be transported to microgravity and stored for periods of time between operations. Biofilms were stored without the reactor bulk fluid to simulate transport of established biofilms into microgravity, while biofilms stored with the reactor bulk fluid simulated the most simplistic storage condition to implement operations for extended periods of nonuse. Dormancy condition did not have an influence on recovery in initial studies with immature biofilms (48 days old), however, a lengthy recovery time was required (20+ days). Bioreactors with fully established biofilms (13 months) were able to recover from a 7-month dormancy period to steady state operation within 4 days (approximately 1 residence cycle). Results indicate a need for future testing on biofilm age and health and further exploration of dormancy length.

Dormancy

Selection of Leafy Green Vegetable Varieties for a Pick-and-Eat Diet Supplement on ISS

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.

Nutrition