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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
NASA NTRS2010

Microbial Anomalies Encountered on the International Space Station

Microorganisms in our living environments are unavoidable. A community of microbes arrived in space with the delivery of the first element of the International Space Station (ISS), attached to hardware and on the bodies of the humans tasked with the initial assembly missions. The risk that microorganisms could cause adverse effects in the health of both the human occupants of the ISS as well as the physical integrity of the station environment and life support systems has been both a driver and a function of engineering and operational controls. Scientists and engineers at NASA have gone to extensive measures to control microbial growth at levels safe for the crewmembers and the spacecraft environment. Many of these measures were initiated with the design of the spacecraft and its systems. Materials used in the ISS were tested for resistance to fungi, such as mold and a paint with a fungus-killing chemical was also used. Controlling the humidity of the air in the Station is also an effective way of discouraging microbe growth. The breathing air is reconditioned by the Environmental Control Life Support System (ECLSS) prior to distribution, utilizing High Efficiency Particulate Air (HEPA) filtration. Requirements restricting the accumulation of water condensate in the air handlers and habitable volume of the ISS were other safeguards added. Water for drinking and food rehydration is disinfected or filtered. A robust in-flight housekeeping regimen for the ISS significantly reduces inappropriate growth of microorganisms and includes a regular cleaning of accessible surfaces with disinfectant wipes. Most of these requirements were suggested by microbiologists to mitigate and possibly prevent many microbiological risks. In addition to these controls, before flight monitoring and analyses of the cabin air, exposed surfaces, water and food, consumables, and crew members are conducted to mitigate microbial risk to the crew and spacecraft. Many microbial risks are much easier to identify and resolve before launch than during space flight. Although the focus has been on prevention of microbiologically related, not all problems can be anticipated. A number of microbial anomalies have occurred on ISS. This paper will discuss the occurrences, root-cause investigations, and mitigation steps taken to remediate the contamination.

Bruce, Rebekah J.↗
NASA NTRSDate not supplied

A Lunar Ground Truth of Microbes that are Integral to Sustaining Biological Life Support Systems

A lunar outpost such as Gateway or a Mars transit vehicle will use a semi-closed to closed loop biological life support system (BLSS) adapted for microgravity and high levels of radiation. A moon habitat will likely exist under partial gravity conditions and sustained levels of high radiation as a semi-closed loop system able to get resources from Earth via Gateway. Additionally, the moon will act as a closed-loop testbed for Mars habitat operations and will employ various elements of in situ resource utilization (ISRU). Perhaps the longest running BLSS collective study to date is that of the European MELiSSA (Micro Ecological Life Support System Alternative) project, a circular life support system, established to gain knowledge on regenerative systems aimed at the highest degree of autonomy to produce food, water and oxygen from mission wastes. This setup has evaluated the use of discrete microbial compartments and a higher plant compartment to carry out the necessary life support functions to support crew. Such compartments utilize thermophilic anaerobes to break down human and inedible plant wastes, photoheterotrophic bacteria that can further metabolize volatile fatty acids, nitrifying bacteria that can convert ammonium to plant and microalgae available nitrates, and photoautotrophic bacteria and higher plants which will convert carbon dioxide to oxygen, purify water, and provide food for human consumption. This being the most advanced BLSS model example to date -aside from the Yuegong-1, Chinese Lunar Palace- with the most defined compartment composition, and the assumption that other BLSSs will build upon such models, it becomes relevant to study the effects of the lunar environment upon the microbes that are integral to a BLSS. In ramping up to advanced stage BLSS systems, relatively simple experiments can be conducted on Artemis missions using continuous culturing and sampling of the model microbes, subsequently subjected to DNA sequencing for mutational analysis monitoring and chemical analysis to assess the sustained ability to carry out their hallmark biochemical processes efficiently. A suite of microbes should be assessed, prioritizing those with the duel capacity to be utilized in an BLSS and the ability to biochemically facilitate ISRU goals, such as the ability to transform the biogeochemistry of Moon or Martian regolith into materials that can support crop growth or extract elements of industrial significance such as aluminum or iron.

Biological Life Support Systems↗
NASA NTRSDate not supplied

Benchmarking Computational Tools for Calling SNPs and Indels in Complex Microbial Populations

The NASA BioNutrients missions seek to understand the suitability of microorganisms for bioproduction during space flight. One topic of interest is the stability of microbial genomes during long-term ambient storage and subsequent rehydration and growth. To address these questions, samples from 8 species were flown to ISS for 5 years of desiccated storage at ambient temperature (Stasis Packs) and 2 species were packaged along with powdered media inside a bioreactor system to allow hydration and growth in microgravity (Production Packs). For both systems, Whole Genome Sequencing (WGS) of the DNA extracted from the returned samples and paired ground controls will be conducted to identify changes in genome stability due to time, storage conditions and growth in space. Across the technical replicates, ground controls, 10 timepoints, and multiple experimental conditions, ~300 samples have been selected for initial analysis with WGS sequencing to 100x coverage. A flexible and resource efficient mutation calling pipeline is needed to process this large dataset and allow for comparisons between species. Many bioinformatics tools for calling Indels and Single Nucleotide Variants (SNVs) are designed for use with pure isolates, where true variations from the reference genome are expected to dominate the reads aligning to the location of mutation. In contrast, DNA from the Stasis Pack (SP) samples was collected directly after recovery from desiccated storage and the Production Pack (PP) samples were collected after fermentation. In this context, reads with mutations are expected to be less frequent than reads that align with the reference genome, as each sample will include multiple lines of cells. Thus, BioNutrients samples are expected to be similar to samples from cancer cell or “pooled” sequencing approaches. In preparation for the analysis of the BioNutrients samples, we have tested three mutation calling tools (GATK for Microbes, BreSeq and DiscoSNP) designed for complex samples. A challenge of validating mutation identification pipelines is a lack of “Ground Truth” datasets, especially for complex samples. To compare these three tools, we sought to identify mutations in pre-existing WGS data collected from populations of Chlamydomonas reinhardtii that were exposed to UV mutagenesis and growth in LEO as part of the Space Algae-1 mission. Here we present a summary of these tools against the analysis originally conducted using the CRISP tool. Critical metrics are compared such as runtime, the number of SNPs, the number and size of Indels, and patterns of transversion and transitions identified by each tool are reported. By sharing these benchmarking results collected in support of the BioNutrients mission, we aim to guide others seeking to identify SNVs in similarly complex microbial samples.

Biology↗
NASA NTRSDate not supplied

Harnessing Synthetic Communities and Microbial Recycling of Space Waste Streams for Biomanufacturing Applications

The long-term habitation of extraterrestrial environments such as the Moon or Mars presents significant challenges including supplying materials to sustain life. Off-world recycling of waste materials into biomanufacturer products may ameliorate this. Current literature highlights the need for efficient waste recycling systems to support the bioproduction of essential materials including foods, pharmaceuticals, and biomaterials. The study herein concerns itself with the investigation of three key aspects: 1) formulating an optimal wastewater media on which to grow recombinant microbes for bioproduction in space, 2) examining the potential for constructing stable and metabolically synergistic synthetic microbial communities for largescale and multi-tiered biomanufacturing, and 3) testing the efficacy of one such bioengineered enzyme, cutinase, on the degradation of PET plastics characteristic of those found in ISS waste as a model for recombinant recycling-based biomanufacturing of mission-critical substrates. Formulation of an optimal wastewater media involved growing several microorganisms on mixtures of synthetic planetary wastes representative of those found in space waste systems, combined with simple carbon sources derived from a physio-chemical CO conversion system to determine their growth potential. Potential synthetic microbial communities were conceptually designed, and their stability and metabolic synergism was evaluated within the context of co-cultures. Cutinase activity assays were utilized to determine the efficacy of bioengineered cutinase on PET plastic degradation. Findings will contribute to optimization of wastewater-based media formulations, data on stable synthetic microbial communities for bioproduction, and effective methods for measuring cutinase-based PET plastic degradation. These outcomes support the development of sustainable waste recycling systems for space habitation and aim to fill gaps in the current literature and proposing innovative solutions for waste recycling in space environments. By leveraging synthetic biology this study seeks to enhance the feasibility of long-term extraterrestrial habitation through sustainable resource management.

Bioprocessing↗
NASA NTRS1996

Significance of Plant Root Microorganisms in Reclaiming Water in CELSS

Since many microorganisms demonstrate the ability to quickly break down complex mixtures of waste and environmental contaminants, examining their potential use for water recycling in a closed environment is appealing. Water contributes approximately 90 percent of the life sustaining provisions in a human space habitat. Nearly half of the daily water requirements will be used for personal hygiene and dish washing. The primary contaminants of the used "gray" water will be the cleansing agents or soaps used to carry out these functions. Reclaiming water from the gray water waste streams is one goal of the NASA program, Controlled Ecological Life Support Systems (CELSS). The microorganisms of plane roots are well documented to be of a beneficial effect to promote plant growth. Most plants exhibit a range of bacteria and fungi which can be highly plant-specific. In our investigations with lettuce grown in hydroponic culture, we identified a microflora of normal rhizosphere. When the roots were exposed to an anionic surfactant, the species diversity changed, based on morphological characteristics, with the numbers of species being reduced from 7 to 2 after 48 hours of exposure. In addition, the species that became dominant in the presence of the anionic surfactant also demonstrated a dramatic increase in population density which corresponded to the degradation of the surfactant in the root zone. The potential for using these or other rhizosphere bacteria as a primary or secondary waste processor is promising, but a number of issues still warrant investigation; these include but are not limited to: (1) the full identification of the microbes, (2) the classes of surfactants the microbes will degrade, (3) the environmental conditions required for optimal processing efficiency and (4) the ability of transferring the microbes to a non-living solid matrix such as a bioreactor.

Bubenheim, David L.↗
NASA NTRS2014

Application of Synthetic Biology to Bioregenerative Life Support for Human Spaceflight

The conversion of carbon dioxide into higher value products is a key challenge for the development of closed-loop life support systems for human space flight. Much of the past research on bioregenerative life support systems has focused on plant growth chambers as a solution for CO2 removal and O2 generation, but photosynthetic microorganisms may also have a role to play in these functions. Cyanobacteria have the advantages of relatively high CO2 fixation rates and fairly well-developed molecular biology tools, allowing for genetic engineering approaches to strain improvement. Manned missions to Mars or other targets beyond low Earth orbit will require advances in the nutritional systems for life support on these longer duration missions. A key challenge will likely be supplementing pre-packaged meals with specific nutrients that will be deficient due to problems in long-term storage or low abundance. Vitamin K is one such nutrient that may be important as a supplement. Production of vitamin K for nutrient supplementation during spaceflight will likely require genetic engineering of microorganisms to increase vitamin titers. A microbial bioreactor system that could efficiently convert CO2 to nutritional supplements would be a valuable component for a future advanced life support system. We are exploring biological systems to determine the feasibility of using bioreactors to convert CO2 to higher-value products. We are examining the performance of photosynthetic bacteria engineered to produce sugars, determining rates of production and reliability. We are also engineering microbes to produce higher titers of vitamin K and other potentially important nutrients. The results of this research will offer demonstrations of potential technologies that could be developed further in the future. This work will also provide valuable information for understanding basic science questions about the use of genetically engineered microbes in the microgravity environment.

Bioregenerative life support↗