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Leverage additive manufacturing to fortify ECLSS (Environmental Control and Life Support Systems) manifolds and other vulnerable components against biofilm growth during dormancy. This project aims to improve dormancy tolerance by eliminating dead legs to reduce stagnate fluid volume and to limit available nutrients, and by printing from inherently biocidal materials.
It should be possible to construct a general equation describing the survival of microbes transferred between Solar System bodies. Such an equation will be useful for constraining the likelihood of transfer of viable organisms between bodies throughout the lifetime of the Solar System, and for refining Planetary Protection constraints placed on future missions. We will discuss the construction of such an equation, present a plan for definition of pertinent factors, and will describe what research will be necessary to quantify those factors. Description: We will examine the case of microbes transferred between Solar System bodies as residents in meteorite material ejected from one body (the "intial body") and deposited on another (the "target body"). Any microbes transferred in this fashion will experience four distinct phases between their initial state on the initial body, up to the point where they colonize the target body. Each of these phases features phenomena capable of reducing or exterminating the initial microbial population. They are: 1) Ejection: Material is ejected from the initial body, imparting shock followed by rapid desiccation and cooling. 2) Transport: Material travels through interplanetary space to the target body, exposing a hypothetical microbial population to extended desiccation, irradiation, and temperature extremes. 3) Infall: Material is deposited on the target body, diminishing the microbial population through shock, mass loss, and heating. 4) Adaptation: Any microbes which survive the previous three phases must then adapt to new chemophysical conditions of the target body. Differences in habitability between the initial and target bodies dominate this phase. A suitable general-form equation can be assembled from the above factors by defining the initial number of microbes in an ejected mass and applying multiplicitive factors based on the physical phenomena inherent to each phase. It should be possible to present the resulting equation in terms of initial ejection mass, ejection shock magnitude, transfer time, initial microbial load and/or other terms and generate graphs defining the number of surviving microbes. The general form of the equation is: x(sub f) = x(sub i) f(sub1) f(sub 2) f(sub 3) f(sub 4) Where x(sub f) is the final number of microbes to survive transfer, x(sub i) is the initial population prior to ejection, and f(sub 1-4) are mortality factors for the four phases described above. Among other considerations, f(sub 1) will vary with respect to impact shock magnitude and f(sub 2) will be time-dependent. Considerable research has been performed to date to quantify the survival rates of various microbes in response to portions of these four phases, both as vegetative cells and/or spores. Results indicate that many species tend to respond differently to the pertinent mortality factors, especially in the case of extremophiles. Therefore, a complete equation will include species-specific responses to the mortality factors.
The Automated Microbial Metabolism Laboratory (AMML) 1971-1972 program involved the investigation of three separate life detection schemes. The first was a continued further development of the labeled release experiment. The possibility of chamber reuse without inbetween sterilization, to provide comparative biochemical information was tested. Findings show that individual substrates or concentrations of antimetabolites may be sequentially added to a single test chamber. The second detection system which was investigated for possible inclusion in the AMML package of assays, was nitrogen fixation as detected by acetylene reduction. Thirdly, a series of preliminary steps were taken to investigate the feasibility of detecting biopolymers in soil. A strategy for the safe return to Earth of a Mars sample prior to manned landings on Mars is outlined. The program assumes that the probability of indigenous life on Mars is unity and then broadly presents the procedures for acquisition and analysis of the Mars sample in a manner to satisfy the scientific community and the public that adequate safeguards are being taken.
During the Antarctica 2000 Expedition, sponsored by the Planetary Studies Foundation, meteorites and ice microbiota were collected from the Patriot Hills, and Thiel Mountains of Antarctica and snow samples were at the South Pole. Psychrophilic and psychrotrophic microbiota were obtained from blue ice, cryoconite and ice-bubble systems. Twenty frozen meteorites were collected using aseptic techniques from the blue ice fields near the Moulton Escarpment of the Thiel Mountains (85 S, 94 W) and from the Morris Moraine of the Patriot Hills (80 S, 81 W) Ellsworth Mountains. These ice and meteorite samples are of potential significance to Astrobiology. They may help refine chemical and morphological biomarkers and refine characteristics of microbial life in one of the harshest environments on Earth. We discuss the Antarctica 2000 Expedition and provide preliminary results of the investigation of the meteorites and ice microbiota recovered.
Planetary Protection (PP) at NASA aims to 1) protect solar system bodies from contamination by Earth life and 2) protect Earth from possible life forms that may be returned from other solar system bodies. It is particularly relevant for missions focused on life detection. PP specialists at MSFC act as subject matter experts to interpret PP requirements and communicate with other NASA centers and/or vendors to ensure compliance. This includes insight and oversight to help projects consider and plan PP implementation during design phases. It also includes physical implementation during build, assembly, and test operations, (i.e. microbial sampling of environments and spacecraft) to help maintain cleanliness. MSFC has also expanded to include PP research which included standing up a fully functional BSL2 microbiology laboratory.
The three main topics of this program as described initially in our May 2003 proposal are: 1) Shock-induced damage and attenuation in planetary materials. 2 ) Shock-induced melting and phase changes. 3) Impact-induced volatilization and vapor speciation of planetary materials Topic 4 has been the subject of a continuing investigation since approximately 1990. On Topic 5, we have a paper in preparation and have submitted a proposal to Astrobiology. 4) Responses of planetary atmospheres to giant impact, 5) Effects of impact-induced shock waves on microbial life
We have designed and built a prototype Supercritical CO? Cleaning (SCC) system at JPL. The key features of the system are: 1) the parts inside a high-pressure vessel can be rotated at high speeds; 2) the same thermodynamic condition is maintained during First-In First-Out flushing to keep solvent power constant; and 3) the boil-off during decompression is induced in a separate vessel downstream. Our goal is to demonstrate SCC's ability to remove trace amounts of microbial and organic contaminants down to parts per billion levels from spacecraft material surfaces for future astrobiology missions. The initial cleaning test results showed that SCC can achieve cleanliness levels of 0.01 microgram/cm(sup 2) or less for hydrophobic contaminants such as dioctyl phthalate and silicone and it is less effective in the removal and inactivation of the hydrophilic bacterial spores as expected. However, with the use of a polar co-solvent, the efficacy may improve dramatically. The same results were obtained using liquid CO?. This opens up the possibility of using subcritical cleaning conditions, which may prove to be more compatible with certain spacecraft hardware.
Microorganisms grow differently in spaceflight than they do on Earth. While numerous factors likely contribute, one ubiquitous feature of the space environment is altered gravity - but because microbes are generally considered too small to detect gravity directly, they likely experience the effects of gravity via changes in their fluid environment, primarily a reduction in convective mixing. This theory is supported by evidence that individual microbial strains experience starvation and acid stress in microgravity. If it is true, then microgravity would also have measurable effects on microbes in mixed communities, because many interspecies interactions involve the exchange of soluble metabolites through their medium (cross-feeding). Specifically, cooperative cross-feeding communities would grow more slowly in microgravity, and this effect should be reproducible in ground-based rotational culture systems (rotating wall vessels, RWVs) designed to simulate the quiescent environment of microgravity. Here we present preliminary results of our efforts to test this hypothesis using using a model system of Eschericia coli and Salmonella enterica that grow only when they can exchange methionine and acetate through the medium. We report on the development of a novel laboratory system that allows us measure growth rates and species ratios in real-time using fluorescence spectrometry as the organisms grow in RWVs. We are able to track growth rates of individual species and the co-culture continuously with high temporal resolution for several hours, a substantial improvement over prior RWV systems that require destructive sampling. We vary the fidelity of the microgravity simulation by varying the rotation rate, and find that rotation rate affects growth rates of the community more than that of individual strains, consistent with the claim that RWVs can simulate a microgravity-like quiescent environment. While more work needs to be done to probe the mechanisms of this effect, we hope that our system will ultimately allow us to generate quantitative predictions about the effects of spaceflight on organisms that are tiny but essential to sustaining long-term human space exploration.
The White House Office of Science and Technology Policy (OSTP) has declared 2023 to be the Year of Open Science following an initiative to democratize scientific knowledge. Simultaneously, new sensor technologies have broadened the experimental space available to bioastronautics research. With these open-science goals and technological advances in mind, we have designed and constructed a data acquisition platform for high-precision, real-time monitoring of liquid culture systems. The vessel rig is fitted with six Atlas Scientific probes (micro pH, electrical conductivity, dissolved oxygen, oxidation-reduction potential, liquid temperature, air CO2) and a custom optical density probe similar to the one on BioSentinel’s BioSensor payload. A custom dielectric spectroscopy probe is also planned. The structure of the vessel is resin 3-D printed on a hobbyist-level machine, reducing the production cost and iteration time by over 60% each while increasing extensibility. Data acquisition and storage is controlled with a standalone C state machine-based program running on a Raspberry Pi 3 Model B. When not running headless, an additional program automatically generates and updates plots for live data visualization. Validation of the rig as a data collection system was performed with a yeast liquid culture experiment. While the vessel rig is currently used for standalone experiments, it can also be used as the base perception unit in a self-driving laboratory (SDL). SDLs are high-throughput data collection systems that employ automation and artificial intelligence to conduct and manage routine experiments. Here, we envision an SDL driven by several vessel rigs in which an automated script compares key results, informing the design of future experiments. A vessel rig SDL would streamline many operations, including 1) strain selection for the Lunar Explorer Instrument for space biology Applications (LEIA) investigation and 2) the study of bioregenerative life support systems (BLSS). Ultimately, the datasets that can now be acquired will provide crucial information for accelerating bioastronautics application development in the era of commercial space.
Since the Viking missions in the mid-1970s, traditional culture-based methods have been used for microbial enumeration by various NASA programs. Viable microbes are of particular concern for spacecraft cleanliness, for forward contamination of extraterrestrial bodies (proliferation of microbes), and for crew health/safety (viable pathogenic microbes). However, a "true" estimation of viable microbial population and differentiation from their dead cells using the most sensitive molecular methods is a challenge, because of the stability of DNA from dead cells. The goal of this research is to evaluate a rapid and sensitive microbial detection concept that will selectively estimate viable microbes. Nucleic acid amplification approaches such as the polymerase chain reaction (PCR) have shown promise for reducing time to detection for a wide range of applications. The proposed method is based on the use of a fluorescent DNA intercalating agent, propidium monoazide (PMA), which can only penetrate the membrane of dead cells. The PMA-quenched reaction mixtures can be screened, where only the DNA from live cells will be available for subsequent PCR reaction and microarray detection, and be identified as part of the viable microbial community. An additional advantage of the proposed rapid method is that it will detect viable microbes and differentiate from dead cells in only a few hours, as opposed to less comprehensive culture-based assays, which take days to complete. This novel combination approach is called the PMA-Microarray method. DNA intercalating agents such as PMA have previously been used to selectively distinguish between viable and dead bacterial cells. Once in the cell, the dye intercalates with the DNA and, upon photolysis under visible light, produces stable DNA adducts. DNA cross-linked in this way is unavailable for PCR. Environmental samples suspected of containing a mixture of live and dead microbial cells/spores will be treated with PMA, and then incubated in the dark. Thereafter, the sample is exposed to visible light for five minutes, so that the DNA from dead cells will be cross-linked. Following this PMA treatment step, the sample is concentrated by centrifugation and washed (to remove excessive PMA) before DNA is extracted. The 16S rRNA gene fragments will be amplified by PCR to screen the total microbial community using PhyloChip DNA microarray analysis. This approach will detect only the viable microbial community since the PMA intercalated DNA from dead cells would be unavailable for PCR amplification. The total detection time including PCR reaction for low biomass samples will be a few hours. Numerous markets may use this technology. The food industry uses spore detection to validate new alternative food processing technologies, sterility, and quality. Pharmaceutical and medical equipment companies also detect spores as a marker for sterility. This system can be used for validating sterilization processes, water treatment systems, and in various public health and homeland security applications.
This report describes the MSFC space station water degradation study (WDS) and presents interim results from the first 24 months of testing. The WDS simulates the stagnant storage of water in distribution lines before the activation of the space station's water processor by storing processed water at ambient temperature in valved sections of 1-in stainless steel and titanium tube. The WDS seeks to determine whether the water quality will degrade unacceptably and whether microbial growth will proceed to an unmanageable extent during extended stagnation. During the first 24 months, significant changes have occurred. Although iodine, which is used as a biocide, was nearly depleted within the first 6 months of testing, microbial growth has been minimal. This report describes the decrease in iodine concentration and the results of microbial and biofilm analyses. Increases in total organic carbon, iodide, chloride, nickel, iron, and chromium concentrations are presented and discussed. The observed increase in conductivity and the decreases in pH and turbidity are also presented. The authors conclude that, with proper preparation, potable water can be stored under stagnant conditions without unmanageable degradation in water quality; a flushing operation and subsequent processing of the degraded water should render the water system ready for use.
Although present Martian surface conditions appear unfavorable for life as we know it, there is compelling geological evidence that the climate of early Mars was much more Earth-like, with a denser atmosphere and abundant surface water. The fact that life developed on the Earth within the first billion years of its history makes it quite plausible that life may have also developed on Mars. If life did develop on Mars, it is likely to have left behind a fossil record. This has led to the development of a new subdiscipline of paleontology, herein termed 'exopaleontology', which deals with the exploration for fossils on other planets. The most important factor enhancing microbial fossilization is the rapid entombment of microorganisms by fine-grained, stable mineral phases, such as silica, phosphate, or carbonate. The oldest body fossils on Earth are preserved in this way, occurring as permineralized cells in fine-grained siliceous sediments (cherts) associated with ancient volcanic terranes in Australia and South Africa. Modern terrestrial environments where minerals may precipitate in the presence of microorganisms include subaerial thermal springs and shallow hydrothermal systems, sub-lacustrine springs and evaporitic alkaline lakes, zones of mineralization within soils where 'hardpans' (e.g. calcretes, silcretes) form, and high latitude frozen soils or ground ice.
NASA has been developing a new spacesuit, called the Exploration Extravehicular Mobility Unit (xEMU) for over a decade. This spacesuit is underdevelopment to support missions to the International Space Station (ISS) and also to the Moon. The xEMU uses controlled evaporative cooling through a membrane to reject waste heat from the crewmember, avionics, and environmental in-leak during autonomous operation. Given the unique mass and volume constraints imposed by the spacesuit application and the opportunities presented by the selection of a given membrane, the xEMU does not possess a separate feedwater loop to feed the evaporation process, rather the feedwater for the evaporation process is fed directly from the transport loop which collects heat from the sources within the suit. This has many benefits including mass/volume reductions from a lower component count within the system but does pose some additional potential constraints on water quality within the system in order to provide a working fluid that can be pumped, filtered, and used in contact with the remainder of the system while both avoiding fouling of the membrane as well as other deleterious effects such as creating conditions for corrosion of wetted surfaces within the transport loop. The discussion will seek to address the main sources of contamination and the applied controls. Sources considered will include: self-generated via materials selection and processes applied, influent working fluid, contamination of the influent working fluid via Ground Support Equipment (GSE) or processing equipment, microbial activity, and environmental influences such as elevated ambient CO2.
Planetary protection (PP) policy seeks to avoid harmful contamination by limiting biological and relevant organic contamination from spacecraft as well as preventing adverse changes to Earth’s biosphere when extraterrestial samples are brought back to Earth. The PP policy at NASA was updated in 2021 (NPR 8715.24) and 2022 (NASA-STD-8719.27) to enable missions by expanding the decades old prescriptive requirements to allow for an option of adopting performance-based requirements that are objectivesdriven, risk-informed and case-assured. In parallel, the final PP knowledge gap workshop was completed representing the international consensus on the key areas to be considered in developing crew PP policy. These knowledge gaps focused on key technology development areas in 1) microbial and human health monitoring, 2) technical and operations needed for contamination control and 3) natural transport of contamination on Mars. As robotic missions start to implement performance-based approaches and research and technology efforts commence to inform crew policy the demand for data quality driven verification and validation in relevant space environments. Examples of the types of testing that is envisioned includes test as you fly validation and verification of decontamination systems in a relevant on-orbit and Mars environment, developing lethality curves of terrestrial organisms to further our understanding of the biocidal impacts of Mars and the space environment, and particle transport model validation and verification. Thus, the PP discipline has identified the need for groundbased space environments to perform preliminary testing as validation and verification of flight systems and to advance the technology readiness level prior to further testing on-orbit or lunar environments to prepare for Mars.
Planetary protection (PP) policy seeks to avoid harmful contamination by limiting biological and relevant organic contamination from spacecraft as well as preventing adverse changes to Earth’s biosphere when extraterrestrial samples are brought back to Earth. The PP policy at NASA was updated in 2021 (NPR 8715.24) and 2022 (NASA-STD-8719.27) to enable missions by expanding the decades old prescriptive requirements to allow for an option of adopting performance-based requirements that are objectives-driven, risk-informed and case-assured. In parallel, the final PP knowledge gap workshop was completed representing the international consensus on the key areas to be considered in developing crew PP policy. These knowledge gaps focused on key technology development areas in 1) microbial and human health monitoring, 2) technical and operations needed for contamination control and 3) natural transport of contamination on Mars. As robotic missions start to implement performance-based approaches and research and technology efforts commence to inform crew policy the demand for data quality driven verification and validation in relevant space environments. Examples of the types of testing that is envisioned includes test as you fly validation and verification of decontamination systems in a relevant on-orbit and Mars environment, developing lethality curves of terrestrial organisms to further our understanding of the biocidal impacts of Mars and the space environment, and particle transport model validation and verification. Thus, the PP discipline has identified the need for ground-based space environments to perform preliminary testing as validation and verification of flight systems and to advance the technology readiness level prior to further testing on-orbit or lunar environments to prepare for Mars.
Fresh, nutritious, palatable produce for crew consumption on long-duration spaceflight missions may provide health-promoting, bioavailable nutrients and enhance the dietary experience. VEG-04A and VEG-04B explored growing leafy greens on the International Space Station using the Veggie Vegetable Production System. Two flight tests with ground controls were conducted in 2019 growing mizuna mustard, where Veggie chambers were set to different red-to-blue-to-green light formulations. Light quality affects plant growth, nutrition, microbiology, and organoleptic characteristics on Earth, and we examined how these vary in microgravity and under different harvest scenarios. Astronauts harvested and weighed mizuna and completed organoleptic evaluations. Flight samples were returned to Earth for nutritional quality and microbial food safety analyses. Yield and chemistry differed between ground and flight samples and light treatments, and bacterial and fungal counts were lower in ground than in flight samples. This research helps increase our understanding of the requirements for growing high-quality crops in spaceflight.
Lunar and martian materials can be processed and used at planetary outposts to reduce the need (and thus the cost) of transportng supplies from Earth. A variety of uses for indigenous, on-site materials have been suggested, including uses as rocket propellants, construction materials, and life support materials. Utilization of on-site resources will supplement Regenerative Life Support Systems (RLSS) that will be needed to regenerate air, water, wastes, and to produce food (e.g., plants) for human consumption during long-duration space missions. Natural materials on the Moon and/or Mars may be used for a variety of RLSS needs including (1) soils or solid-support substrate for plant growth, (2) sources for extraction of essential plant-growth nutrients, (3) sources of O2, H2, CO2, and water, (4) substrates for microbial populations in the degradation of wastes, and (5) shielding materials surrounding outpost structures to protect humans, plants, and microorganisms from radiation. In addition to the regolith, the martian atmosphere will provide additional resources at a Mars outpost, including water, CO2 and other atmospheric gases.
The ongoing search for life on other worlds and the prospects of eventual human exploration of the Moon and Mars indicate the need for new ethical guidelines to direct our actions as we search and how we respond if we discover microbial life on other worlds. Here we review how life on other worlds presents a novel question in environmental ethics. We propose a principle of protecting and expanding the richness and diversity of life as the basis of an ethic for astrobiology research and space exploration. There are immediate implications for the operational policies governing how we conduct the search for life on Mars and how we plan for human exploration throughout the Solar System.