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At least 55 records · Page 3

Veggie ISS Validation Test Results and Produce Consumption

The Veggie vegetable production system flew to the International Space Station (ISS) in the spring of 2014. The first set of plants, Outredgeous red romaine lettuce, was grown, harvested, frozen, and returned to Earth in October. Ground control and flight plant tissue was sub-sectioned for microbial analysis, anthocyanin antioxidant phenolic analysis, and elemental analysis. Microbial analysis was also performed on samples swabbed on orbit from plants, Veggie bellows, and plant pillow surfaces, on water samples, and on samples of roots, media, and wick material from two returned plant pillows. Microbial levels of plants were comparable to ground controls, with some differences in community composition. The range in aerobic bacterial plate counts between individual plants was much greater in the ground controls than in flight plants. No pathogens were found. Anthocyanin concentrations were the same between ground and flight plants, while antioxidant and phenolic levels were slightly higher in flight plants. Elements varied, but key target elements for astronaut nutrition were similar between ground and flight plants. Aerobic plate counts of the flight plant pillow components were significantly higher than ground controls. Surface swab samples showed low microbial counts, with most below detection limits. Flight plant microbial levels were less than bacterial guidelines set for non-thermostabalized food and near or below those for fungi. These guidelines are not for fresh produce but are the closest approximate standards. Forward work includes the development of standards for space-grown produce. A produce consumption strategy for Veggie on ISS includes pre-flight assessments of all crops to down select candidates, wiping flight-grown plants with sanitizing food wipes, and regular Veggie hardware cleaning and microbial monitoring. Produce then could be consumed by astronauts, however some plant material would be reserved and returned for analysis. Implementation of this plan is a step toward developing pick-and-eat food production to supplement the packaged diet on ISS and for future exploration missions where plants could make up a larger portion of the diet. Supported by NASA Space Biology Program.

Food Safety↗

A Prototype Tool for Assessing Forward Contamination from Space Suits

Crewed exploration missions dramatically increase the capability for large-scale sample collection and return activities, but they also increase the possibility and likelihood of forward contamination. Systematic research on forward contamination from uncrewed spacecraft has steadily progressed since the Viking missions, but parallel research on contamination from space suits has not. Current space suits have leakage rates as high as 100 cc gas/min., but it is unclear how many or what types of microbes are exiting the suits along with this gas. The Human Forward Contamination Assessment team at NASA's Johnson Space Center (JSC) has developed a prototype swab tool that is capable of maintaining sterility during pressure changes associated with entering and exiting vacuum. The primary objective of recent Extravehicular Activity (EVA) Swab Kit testing is to characterize the type of microorganisms typically found on spacesuit external surfaces under suit differential pressure conditions. Most human-associated microorganisms can fit through a 0.5 to 1.0 μm gap. Understanding potential leak paths will inform future hardware design decisions. Knowing which types of microorganisms may leak from EVA suits provides a basis for subsequent studies to characterize the viability of those organisms under destination conditions, as well as how far they might spread through natural or human-influenced processes. The results of EVA suit molecular microbial community analyses will inform NASA exploration mission operations and hardware design, and help close Strategic Knowledge Gap B5, Forward Contamination to Mars.

Rucker, M.↗

Development of Computational Environmental Microbiome Workflows for the Laboratory and the International Space Station

Identification of microorganisms in the spaceflight environment is critical for crew health risk assessment on the International Space Station (ISS). Since 2017, nanopore sequencing technology has been used to support thein situ identification of microbial species during spaceflight. Beginning in 2018, a culture-independent, swab-to-sequencer method was implemented onboard the ISS to provide a more thorough insight of the ISS microbiome. Eliminating microbial culture enables identification of difficult-to-culture organisms, reduces risks associated with potentially pathogenic cultures, and could significantly reduce the time from sample-to-answer. However, this molecular-based approach generates large metagenomic datasets that require substantial computational resources for analysis. To process nanopore-generated sequencing data, the JSC Microbiology Laboratory established a bioinformatics workflow on Amazon EC2 under the security guidance of the NASA Science Managed Cloud Environment (SMCE).This resource allows for the development, testing, and accessing of computational tools for processing large and complex datasets. The work described here will address the downlinking of data from the ISS, the automated pipeline developed to identify targeted bacterial and fungal organisms, and the time from sampling onboard to microbial identification. The pipelines have been enhanced to address high and low biomass samples using optimization based on sample source (air, water, or surface) and type of collection (filter, colony, or swab).The resulting microbiome data can be assessed beyond microbial identifications to gain understanding toward population changes over time, potential selective environmental pressures, and evaluating correlations with a wide range of additional data sets. Metagenome analysis pipelines in development could allow for simultaneous identification of microbial species, gene function, and gene pathways present in the environment. Beyond the ground processing, the developed analysis pipeline is currently deployed onboard the ISS to allow for near real-time assessments of the ISS microbiome. This study serves as a critical foundation for exploration missions, where rapid microbiome analyses will be required.

G. Marie Sharp↗

Culture-Independent Fungal Profiling for the International Space Station using Nanopore Sequencing: Method Development

Microbial monitoring of the International Space Station (ISS) environment is a crew health requirement that encompasses both bacterial and fungal identification. To achieve this currently, culture-based methods are used for sample collection, and these samples must be returned to the laboratory for analysis. The use of culture and the need for sample return to Earth results in a bias toward culturable organisms and causes a significant delay between sample collection and delivery of final data (weeks to months), respectively. Recently, advancements in molecular technology have aided a broad range of applications, including medical, industrial, and basic sciences. Additionally, increases in portability and ease-of-use of molecular platforms have provided point-of-use capabilities demonstrated by the miniPCR thermal cycler (miniPCR bio) and the MinION sequencer (Oxford Nanopore Technologies). Together, these devices have been applied to, and validated for, the identification of bacteria onboard the ISS. Building on this work, we have developed a spaceflight-compatible fungal workflow. Molecular-based fungal analysis is complicated by low biomass, difficult-to-lyse spores, debate regarding the region for taxonomic assignment, and the lack of bioinformatic pipelines and reference databases. To overcome these difficulties, primers yielding an ~ 2 Kb amplicon were validated against a wide range of ISS fungal isolates. The current spaceflight library preparation was substantially optimized, a bioinformatic pipeline was created, and refinements to the UNITE database were implemented. To compare this optimized method to the current culture-based standard, 30 sample sets (60 total swabs, two swabs held in tandem) were evaluated. Parallel fungal profiles were obtained between the two methods, with the culture-independent method revealing increased diversity. The addition of this method to the already established bacterial process fulfills the crew health identification requirement. Moreover, the implementation of this method onboard ISS will enhance our understanding of its unique fungal microbiome.

Hang N. Nguyen↗

VEG-05 Tomato Crop Testing on the International Space Station

Production of fresh, nutritious, and tasty produce for astronauts during spaceflight may provide health-promoting, bioavailable nutrients, enhance the dietary experience, and reduce menu fatigue as we move into longer-duration missions. Growing and caring for plants may also reduce the psychological stresses associated with spaceflight and enhance connection to Earth. A diversity of crops will be required to provide nutrition, variety, and resiliency, however requirements for consistent growth under spaceflight environmental conditions remain unclear. VEG-05 is part of a series of experiments with pick-and-eat salad crops to better define best practices for crop production in space. VEG-05 and predecessor experiments VEG-04A and VEG-04B, grew salad crops in the Veggie vegetable production facilities on the International Space Station using different lighting treatments. In VEG-05 we grew ‘Red Robin’ dwarf cherry tomatoes under two different red: blue lighting spectra. Light can impact the growth habit, yield, nutritional composition, microbial levels, and even flavor attributes within crops, and our goal was to assess these characteristics in ‘Red Robin’ tomatoes during VEG-05. Considerable pre-flight verification testing was performed prior to launch in Nov. 2022. Prior to the flight and ground experiments, lighting in both Veggie units on ISS was measured and lights were standardized between flight and ground hardware. VEG-05 flight operations ran between December 2022 and March 2023, with a ground control initially 48 hours delayed. Due to environmental challenges of very low humidity during the first week of the experiment, seed germination was low for both flight and ground plants. The flight experiment ultimately had 4 - 5 plants per treatment out of the planned 6 plants, but the initial ground control had only three plants in one treatment and none in the other, so this ground control was restarted at the beginning of Feb. 2023 and ran through May, with successful growth of all 12 plants. Both flight and ground control ran 100 days, with harvests of fruit at day 83, day 90, and day 100. Flight plants had uneven growth, and following the early drying events, excess water was frequently observed, which led to a variety of plant stress responses including uneven plant growth, excess adventitious root formation, flower and fruit abortion, and visible microbial growth. In total, from the five surviving red-rich lighted plants, only 5 ripe fruit were produced, and from the four surviving blue-rich lighted plants, 10 fruit were produced with only 6 of these ripe by day 100. Because of the small fruit number and the unsatisfactory growth, crew members were not allowed to consume the tomatoes, and all fruit, as well as large branches with leaves, samples of the adventitious roots, two plant rooting pillows from each treatment, microbial sampling swabs, and some water samples were returned for analysis. Because of the small sample sizes and factors affecting growth on the ISS, objectives of assessing light quality effect (red: blue light treatments) will not be achieved. Revised objectives of this study include to compare stressed flight plants with normal ground plants to determine the impact of plant overwatering stress in space on food safety and the plant microbial community, to determine nutrient content changes in fruit and leaves from stressed plants, and to evaluate stress metabolism changes in returned tissue by transcriptomic analysis. Postflight analysis is underway with the following analyses being conducted: A. culturable microbiology and food safety as well as molecular microbial community analysis of 1. ripe fruit, 2. leaves, stems, and adventitious roots, 3. pillow components (roots, wicks, and substrates), 4. swabs, and 5. water samples from root mats before and after growth. B. transcriptomics of leaf tissue and adventitious roots, and C. elemental analysis of leaf tissue. If sufficient tissue remains elemental analyses will also be conducted on fruit. While not generating the desired information on spaceflight growth responses of healthy crops, our team is hopeful that these analyses will shed light on tomato responses to stress in this environment as plant overwatering stress is a mission-relevant condition that could occur in future space crop growth systems. This research was co-funded by the Human Research Program and Space Biology (MTL#1075) in the ILSRA 2015 NRA call.

Gioia D. Massa↗

The Future of in-Situ Sequencing-Based Microbial Monitoring: Development of a Shelf-Stable Method for Artemis and Beyond

Microbial monitoring onboard the International Space Station (ISS) is essential for assessing the efficiency of the Environmental Control and Life Support Systems (ECLSS) and providing insight into potential risk to both crew and spacecraft. Historically, this monitoring required the need to culture organisms onboard, return these cultures to Earth, and then complete the identifications, a process that would take months. Over the past decade, and through numerous payloads, advances in molecular biology have enabled in-flight microbial identifications using nanopore sequencing. The swab-to-sequencer method resulting from these efforts was transitioned from research to operations for microbial monitoring under the Crew Health Care Systems (CHeCS) BioMole. Collectively, these accomplishments have propelled the swab-to-sequencer method to be selected as the Microbial Surface Monitor (MSM) for Gateway, as well as a payload on Artemis IV. However, the lack of cold stowage availability for Artemis requires modifications to the entire method due to the thermal instability of the reagents required for sample preparation. To achieve this, new development, optimization, and validations were undertaken. Key considerations included enzyme concentration, buffer compatibility, and equal or enhanced sensitivity and specificity. At each step, thorough side-by-side comparisons with the current ISS method were performed. The development of a robust shelf-stable method will ensure continued sequencing-based microbial monitoring for Artemis and beyond, providing data in near real-time, enhancing risk response time, and yielding clear insight into the microbiome of spacecraft.

Christian G Mena↗

Automatic agar tray inoculation device

Automatic agar tray inoculation device is simple in design and foolproof in operation. It employs either conventional inoculating loop or cotton swab for uniform inoculation of agar media, and it allows technician to carry on with other activities while tray is being inoculated.

Wilkins, J. R.↗

Services provided in support of the planetary quarantine requirements

Results are presented of laboratory experiments conducted on the thermal resistance of naturally occurring airborne spores and microbiological examinations of space hardware using long-term slit samplers and rodac plate and swab-rinse methods of sampling environmental surfaces.

Favero, M. S.↗

Release of Bacterial Spores from the Inner Walls of a Stainless Steel Cup Subjected to Thermal Stresses and Mechanical Shock

The release and fallout of particulates from surfaces afforded thermal or impact stress is of concern for control of contamination of Mars from planetary landing vehicles. A metal vessel contaminated by aerosols of spores was used as a model system and the fallout of spores as affected by various mechanisms was examined. Thermal stresses simulating those expected on the Mars lander dislodged approximately .01% of the aerosol deposited surface burden as did a landing shock of 8 to 10G deceleration. Spores imprinted by finger or swab contact yielded similar results. In all cases where repeated cycling of temperature, motion, or shock were employed the majority of fallout occurred in the first cycle. Particles released from the surface were predominantly in the size range 1 to 5 microns.

Wolochow, H.↗

Automatic surface inoculation of agar trays.

Description of a machine and technique for the automatic inoculation of a plastic tray containing agar media with a culture, using either a conventional inoculation loop or a cotton swab. The design of the machine is simple, it is easy to use, and it relieves the operator from the manual task of streaking cultures. The described technique makes possible the visualization of the overall qualitative and, to some extent, quantitative relationships of various bacterial types in a sample tested.

Wilkins, J. R.↗

The Orbiting Primate Experiment (OPE)

Instrumentation and life support systems are described for an experiment to determine the physiological effects of long term space flight on unrestrained, minimally instrumented rhesus macaques flown in orbit for periods up to six months or one year. On return from orbit, vestibular, cardiovascular, and skeletal muscle function will be tested. Blood chemistry and hematological studies will be conducted as well as tests of the immunological competence of selected animals. Nasal, rectal, and throat swabs will be used for bacterial and viral studies, and histopathological and histochemical investigations will be be made of all organs using light and electron microscopy. The experiment is being considered as a payload for the biomedical experiment scientific satellite.

Bourne, G. H.↗

Medical support and technology for long-duration space missions

The current philosophy and development directions being taken towards realization of medical systems for use on board space stations are discussed. Data was gained on the performance of physical examinations, venipuncture and blood flow, blood smear and staining, white blood cell differential count, throat culture swab and colony count, and microscopy techniques during a 28-day period of the Skylab mission. It is expected that the advent of Shuttle flights will rapidly increase the number of persons in space, create a demand for in-space rather than on-earth medical procedures, and necessitate treatments for disorders without the provision for an early return to earth. Attention is being given to pressurized environment and extravehicular conditions of treatment, the possibilities of the use of the OTV for moving injured or ill crewmembers to other space stations, and to isolation of persons with communicable diseases from station crews.

Furukawa, S.↗

Experimental and theoretical investigation of passive damping concepts for member forced and free vibration

Potential passive damping concepts for use in space structures are identified. The effectiveness of copper brush, wool swab, and silly putty in chamber dampers is investigated through natural vibration tests on a tubular aluminum member. The member ends have zero translation and possess partial rotational restraints. The silly putty in chamber dampers provide the maximum passive damping efficiency. Forced vibration tests are then conducted with one, two, and three damper chambers containing silly putty. Owing to the limitation of the vibrator used, the performance of these dampers could not be evaluated experimentally until the forcing function was disengaged. Nevertheless, their performance is evaluated through a forced dynamic finite element analysis conducted as a part of this investigation. The theoretical results based on experimentally obtained damping ratios indicate that the passive dampers are considerably more effective under member natural vibration than during forced vibration. Also, the maximum damping under forced vibration occurs at or near resonance.

Razzaq, Zia↗

Milder Etchant For Penetrant Inspection

New etching solution for chemical penetrant inspection of Inconel(R) 718 castings and weldments. Etchant does not introduce artifacts mistaken for defects. Applied by swabbing or by immersion. Used to detect unwanted residues of Nioro(R) (or equivalent) gold brazing alloy on type 347 stainless steel.

O'Tousa, Joseph E.↗

Preflight and postflight microbiological results from 25 space shuttle crews

Clinical-microbiological investigations are an important aspect of the crew health stabilization program. To ensure that space crews have neither active nor latent infections, clinical specimens, including throat and nasal swabs and urine samples, are collected at 10 days (L-10) and 2days (L-2) before launch, and immediately after landing (L+0). All samples are examined for the presence of bacteria and fungi. In addition, fecal samples are collected at L-10 and examined for bacteria, fungi and parasites. This paper describes clinical-microbiological findings from 144 astronauts participating in 25 Space Shuttle missions spanning Space Transportation System (STS)-26 to STS-50. The spectrum of microbiological findings from the specimens included 25 bacterial and 11 fungal species. Among the bacteria isolated most frequently were Staphylococcus aureus, Enterobacter aerogenes, Enterococcus faecalis, Escherichia coli, Proteus mirabilis and Streptococcus agalactiae. Candida albicans was the most frequently isolated fungal pathogen.

Pierson, Duane L.↗

Device For Sampling Surface Contamination

Specially designed cotton swab suitable for use in clean room. Contamination-sampling device improved version of basic cotton-tipped wooden applicator, designed for extreme cleanliness and to enhance utility for sampling according to exacting specifications. Device does not shed cotton and wood fibers.

Delgado, Felix A.↗

Evaluations of an Enhanced Total Hydrocarbon Analyzer With Complex Mixtures of Volatile Organic Compounds

From the earliest manned missions, the volatile organic compound (VOC) content of spacecraft air has been a concern because of a much greater potential for contamination than air in most terrestrial settings. First, the volume of air is small compared to the mass of man- made materials comprising the interior furnishings of the spacecraft. These man-made materials offgas VOCs trapped during manufacture. Second, the nitrogen fraction of the air is recycled. Any VOCs not scrubbed out with charcoal filters or aqueous condensate (mainly water expired by the crew) will accumulate in the air. Third, the crew emits metabolic VOCs. Fourth, experimental payloads can also offgas or accidentally release a VOC; in fact a major organic constituent of the atmosphere is the disinfectant isopropanol released from swabs used in medical experiments.

Cross, John H.↗