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NASA NTRS2010

Microbial Challenge Testing of Single Liquid Cathode Feed Water Electrolysis Cells for the International Space Station (ISS) Oxygen Generator Assembly (OGA)

The International Space Station (ISS) Oxygen Generator Assembly (OGA) operational performance may be adversely impacted by microbiological growth and biofilm formation over the electrolysis cell membranes. Biofilms could hinder the transport of water from the bulk fluid stream to the membranes and increase the cell resistance resulting in higher cell voltages and a shorter cell life. A microbial challenge test was performed on duplicate single liquid cathode feed electrolyzer cells to evaluate operational performance with increasing levels of a mixture of five bacteria isolated from ISS and Space Shuttle potable water systems. Baseline performance of the single water electrolysis cells was determined for approximately one month with deionized water. Monthly performance was also determined following each inoculation of the feed tank with 100, 1000, 10,000 and 100,000 cells/ml of the mixed suspension of test bacteria. Water samples from the feed tank and recirculating water loops for each cell were periodically analyzed for enumeration and speciation of bacteria and total organic carbon. While initially a concern, this test program has demonstrated that the performance of the electrolysis cell is not adversely impacted by feed water containing the five species of bacteria tested at a concentration measured as high as 1,000,000 colony forming units (CFU)/ml. This paper presents the methodologies used in the conduct of this test program along with the performance test results at each level of bacteria concentration.

Diderich, Greg S.↗

NASA NTRS2012

Precision Subsampling System for Mars Surface Missions

The ability to analyze heterogeneous rock samples at fine spatial scales would represent a powerful addition to our planetary in situ analytical toolbox. This is particularly true for Mars, where the signatures of past environments and, potentially, habitability are preserved in chemical and morphological variations across sedimentary layers and among mineral pr.ases in a given rock specimen. On Earth, microbial life often associates with surfaces at the interface of chemical nutrients, and ultimately retains sub-millimeter to millimeter-scale layer confinement in fossilization. On Mars, and possibly other bodies, trace chemical markers (elemental, organic/molecular, isotopic, chiral, etc.) and fine-scale morphological markers (e.g., micro-fossils) may he too subtle, degraded, or ambiguous to be detected, using miniaturized instrumentation, without some concentration or isolation. This is because (i) instrument sensitivity may not be high enough to detect trace markers in bulk averages; and (ii) instrument s~lectiviry may not be sufficient to distinguish such markers from interfering/counteracting signals from the bulk. Moreover from a fundamental chemostratigraphic perspective there would be a great benefit to assessing specific chemical and stable isotopic gradients, over millimeter-to-centimeter scales and beyond, with higher precision than currently possible in situ. We have developed a precision subsampling system (PSS) that addresses this need while remaining relatively flexible to a variety of instruments that may take advantage of the capability on future missions. The PSS is relevant to a number of possible lander/rover missions, especially Mars Sample Return. Our specific PSS prototype is undergoing testing under Mars ambient conditions, on a variety of natural analog rocks and rock drill cores, using a set of complementary flight-compatible measurement techniques. The system is available for testing with other contact instruments that may benefit from precision sampling.

Mahaffy, P. R.↗

NASA NTRS2002

A geostatistical analysis of small-scale spatial variability in bacterial abundance and community structure in salt marsh creek bank sediments

Small-scale variations in bacterial abundance and community structure were examined in salt marsh sediments from Virginia's eastern shore. Samples were collected at 5 cm intervals (horizontally) along a 50 cm elevation gradient, over a 215 cm horizontal transect. For each sample, bacterial abundance was determined using acridine orange direct counts and community structure was analyzed using randomly amplified polymorphic DNA fingerprinting of whole-community DNA extracts. A geostatistical analysis was used to determine the degree of spatial autocorrelation among the samples, for each variable and each direction (horizontal and vertical). The proportion of variance in bacterial abundance that could be accounted for by the spatial model was quite high (vertical: 60%, horizontal: 73%); significant autocorrelation was found among samples separated by 25 cm in the vertical direction and up to 115 cm horizontally. In contrast, most of the variability in community structure was not accounted for by simply considering the spatial separation of samples (vertical: 11%, horizontal: 22%), and must reflect variability from other parameters (e.g., variation at other spatial scales, experimental error, or environmental heterogeneity). Microbial community patch size based upon overall similarity in community structure varied between 17 cm (vertical) and 35 cm (horizontal). Overall, variability due to horizontal position (distance from the creek bank) was much smaller than that due to vertical position (elevation) for both community properties assayed. This suggests that processes more correlated with elevation (e.g., drainage and redox potential) vary at a smaller scale (therefore producing smaller patch sizes) than processes controlled by distance from the creek bank. c2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Non-NASA Center↗

NASA NTRS1977

Engineering and simulation of life science Spacelab experiments

Approaches to the planning and realization of Spacelab life sciences experiments, which may involve as many as 16 Space Shuttle missions and 100 tests, are discussed. In particular, a Spacelab simulation program, designed to evaluate problems associated with the use of live animal specimens, the constraints imposed by zero gravity on equipment operation, training of investigators and data management, is described. The simulated facility approximates the hardware and support systems of a current European Space Agency Spacelab model. Preparations necessary for the experimental program, such as crew activity plans, payload documentation and inflight experimental procedures are developed; health problems of the crew, including human/animal microbial contamination, are also assessed.

Bush, B.↗

NASA NTRS1990

Environmental control and life support systems

Viewgraphs on Environmental Control and Life Support Systems (ECLSS) for Space Station Freedom are presented. Topics covered include: crew generated wastes processing and reclamation; water reclamation - pre- and post-treatment; simplified waste water processing; improved trace contaminant removal; and real time microbial analysis.

Ray, Charles D.↗

NASA NTRS2012

Next Generation Microbiology Requirements

As humans continue to explore deep into space, microorganisms will travel with them. The primary means to mitigate the risk of infectious disease are a combination of prudent spacecraft design and rigorous operational controls. The effectiveness of these methods are evaluated by microbiological monitoring of spacecraft, food, water, and the crew that is performed preflight, in-flight, and post-flight. Current NASA requirements associated with microbiological monitoring are based on culture-based methodology where microorganisms are grown on a semi-solid growth medium and enumerated. Subsequent identification of the organisms requires specialized labor and large equipment, which historically has been performed on Earth. Requirements that rely strictly on culture-based units limit the use of non-culture based monitoring technology. Specifically, the culture-based "measurement criteria" are Colony Forming Units (CFU, representing the growth of one microorganism at a single location on the agar medium) per a given volume, area, or sample size. As the CFU unit by definition is culture-based, these requirements limit alternative technologies for spaceflight applications. As spaceflight missions such as those to Mars extend further into space, culture-based technology will become difficult to implement due to the (a) limited shelf life of the culture media, (b) mass/volume necessary to carry these consumables, and (c) problems associated with the production of biohazardous material in the habitable volume of the spacecraft. In addition, an extensive amount of new knowledge has been obtained during the Space Shuttle, NASA-Mir, and International Space Station Programs, which gave direction for new or modified microbial control requirements for vehicle design and mission operations. The goal of this task is to develop and recommend a new set of requirements for vehicle design and mission operations, including microbiological monitoring, based upon "lessons learned" and new technology. During 2011, this study focused on evaluating potable water requirements by assembling a forum of internal and external experts from NASA, other federal agencies, and academia. Key findings from this forum included: (1) Preventive design and operational strategies should be stringent and the primary focus of NASA's mitigation efforts, as they are cost effective and can be attained with conventional technology. (2) Microbial monitoring hardware should be simple and must be able to measure the viability of microorganisms in a sample. Multiple monitoring technologies can be utilized as long as at the microorganisms being identified can also be confirmed as viable. (3) Evidence showing alterations in the crew immune function and microbial virulence complicates risk assessments and creates the need for very conservative requirements. (4) One key source of infectious agents will always be the crew, and appropriate preventative measures should be taken preflight. (5) Water systems should be thoroughly disinfected (sterilized if possible) preflight and retain a residual biocide throughout the mission. Future forums will cover requirements for other types of samples, specifically spaceflight food and environmental samples, such as vehicle air and vehicle and cargo surfaces. An interim report on the potable water forum has been delivered to the Human Research Program with a final report on the recommendations for all sample types being delivered in September 2013.

Ott, C. M.↗

NASA NTRS1985

Microbial ecology of extreme environments: Antarctic yeasts and growth in substrate-limited habitats

The high, dry valleys of the Ross Desert of Antarctic, characterized by extremely low temperatures, aridity and a depauperate biota, are used as an analog of the postulated extreme climates of other planetary bodies of the Solar System to test the hypothesis that if life could be supported by Ross, it might be possible where similar conditions prevail. The previously considered sterility of the Ross Desert soil ecosystem has yielded up an indigenous yeast, Cryptoccus vishniacci, which is able to resist the extremes of cold, wet and dry freezing, and long arid periods, while making minimal nutritional demands on the soil.

Vishniac, H. S.↗

NASA NTRS1965

Study for control of microbial growth in manned spacecraft Phase I report, 30 Oct. 1964 - 30 Jan. 1965

Microorganism growth control in manned spacecraft environment

MANNED SPACECRAFT↗

NASA NTRS1980

Literature review of human microbes' interaction with plants

Human carried microorganisms, which cannot practically be excluded from human supporting agricultural systems of extra terrestrial stations, are considered. These microorganisms damage the plants on which the people depend for oxygen and food. The inclusion of carefully screened or constructed, but more or less normal, phylloplane and rhizosphere microbial communities is studied.

Maguire, B., Jr.↗

NASA NTRS1987

Plant Growth Module (PGM) conceptual design

The Plant Growth Module for the Controlled Ecological Life Support System (CELSS), designed to answer basic science questions related to growing plants in closed systems, is described functionally with artist's conception drawings. Subsystems are also described, including enclosure and access; data acquisition and control; gas monitor and control; heating, ventilation, and air conditioning; air delivery; nutrient monitor and control; microbial monitoring and control; plant support and nutrient delivery; illumination; and internal operations. The hardware development plan is outlined.

Schwartzkopf, Steven H.↗

NASA NTRS2005

Hydrazine inactivates bacillus spores

Planetary Protection places requirements on the maximum number of viable bacterial spores that may be delivered by a spacecraft to another solar system body. Therefore, for such space missions, the spores that may be found in hydrazine are of concern. A proposed change in processing procedures that eliminated a 0.2 um filtration step propmpted this study to ensure microbial contamination issue existed, especially since no information was found in the literature to substantiate bacterial spore inactivation by hydrazine.

Bacillus↗

NASA NTRS2011

Microbial Challenge Testing of Single Liquid Cathode Feed Water Electrolysis Cells for the International Space Station (ISS) Oxygen Generator Assembly (OGA)

The International Space Station (ISS) Oxygen Generator Assembly (OGA) operational performance may be adversely impacted by microbiological growth and biofilm formation over the electrolysis cell membranes. Biofilms could hinder the transport of water from the bulk fluid stream to the membranes and increase the cell concentration overpotential resulting in higher cell voltages and a shorter cell life. A microbial challenge test was performed on duplicate single liquid-cathode feed water electrolysis cells to evaluate operational performance with increasing levels of a mixture of five bacteria isolated from ISS and Space Shuttle potable water systems. Baseline performance of the single water electrolysis cells was determined for approximately one month with deionized water. Monthly performance was also determined following each inoculation of the feed tank with 100, 1000, 10,000 and 100,000 cells/ml of the mixed suspension of test bacteria. Water samples from the feed tank and recirculating water loops for each cell were periodically analyzed for enumeration and speciation of bacteria and total organic carbon. While initially a concern, this test program has demonstrated that the performance of the electrolysis cell is not adversely impacted by feed water containing the five species of bacteria tested at a concentration measured as high as 1,000,000 colony forming units (CFU)/ml. This paper presents the methodologies used in the conduct of this test program along with the performance test results at each level of bacteria concentration.

Roy, Robert J.↗

NASA NTRS2017

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

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

International Space Station↗

NASA NTRS2018

Plant Growth Research for Future Food Production - Seed Microbiome Project

Humanity is currently on the precipice of a new era: one where human civilization is no longer bound to the confines of a single planet. Now, organizations like the National Aeronautics and Space Administration (NASA) have extended their areas of research and are beginning to focus not just on getting humans into space, but also to keep them safe, healthy, and sane. This focus falls under the Human Research Program (HRP). A focus of this program is "dedicated to discovering the best methods and technologies to support safe, productive human space travel" [1]. For "safe and productive space travel" to occur, astronauts must have proper nutrition [1]. While sending up large amounts of dried and packaged food with every shuttle might work fairly well in the short term, when only a few people need to be fed, it will not be sustainable, especially as NASA looks toward longer space journeys beyond the Earth's orbit. Research into this area falls under Advanced Life Support (ALS), whose mission is to develop regenerative life support systems to support future NASA long-duration missions [2]. This would involve growing crops in space to supplement astronaut diets [2]. An important, yet often overlooked, part of growing crops in any environment is the microbial organisms that inhabit the plants' microbiomes. The Seed Microbiome Project aims to investigate the microbial presence throughout the life stages of three crops, Mizuna Mustard, 'Outredgeous' Red Romaine Lettuce, and 'Red Robin' Tomato, that have either been or will be grown on the ISS.

Birtell, Eva↗

NASA NTRSDate not supplied

Microbiology of Human Spacecraft Environments

Microbiology and Space Missions - Astronaut health - Spaceflight foods - Vehicle habitat - Life support systems - Trash and human waste containment - Astronaut hygiene areas - Planetary protection - How do we protect the crew and extraterrestrial environments in our search for life on other planets? These priority areas for microbiology drive vehicle design, microbial monitoring, and operational research efforts

C. Mark Ott↗

NASA NTRSDate not supplied

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

Many microbial pathogen shave repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in space flight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Herein, we present data from one of these pathogens, the foodborne bacterium, S. enterica Enteritidis, which is closely related to S. enterica Typhimurium. Phenotypes evaluated included growth profiles, environmental stress responses(acid, oxidative, bile, and thermal stresses),and in vitro colonization of3-D biomimetic cultures of human intestinal tissue containing immune cells. Transcriptomic profiling and virulence studies are ongoing. We show that S. Enteritidis exhibited key alterations in pathogenic responses to LSMMG culture that suggest increased infection risk, including several responses which were different from those observed in the closely related pathovar S. Typhimurium. This information will provide critical mechanistic insight into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C M Ott↗

NASA NTRS2024

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

C.M. Ott↗

NASA NTRSDate not supplied

Spaceflight-Induced Changes in Microbial Virulence and the Impact to the Host Immune Response

Many microbial pathogens have repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in spaceflight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. The information to date is providing a better understanding into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.

C M Ott↗