Search NASA⌕ Search

SEARCH · Search NASA

Results for “Microbial biomass”

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.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

105 records · Page 6

Global Partitioning of NOx Sources Using Satellite Observations: Relative Roles of Fossil Fuel Combustion, Biomass Burning and Soil Emissions

This document contains the following abstract for the paper "Global partitioning of NOx sources using satellite observations: Relative roles of fossil fuel combustion, biomass burning and soil emissions." Satellite observations have been used to provide important new information about emissions of nitrogen oxides. Nitrogen oxides (NOx) are significant in atmospheric chemistry, having a role in ozone air pollution, acid deposition and climate change. We know that human activities have led to a three- to six-fold increase in NOx emissions since pre-industrial times, and that there are three main surface sources of NOx: fuel combustion, large-scale fires, and microbial soil processes. How each of these sources contributes to the total NOx emissions is subject to some doubt, however. The problem is that current NOx emission inventories rely on bottom-up approaches, compiling large quantities of statistical information from diverse sources such as fuel and land use, agricultural data, and estimates of burned areas. This results in inherently large uncertainties. To overcome this, Lyatt Jaegle and colleagues from the University of Washington, USA, used new satellite observations from the Global Ozone Monitoring Experiment (GOME) instrument. As the spatial and seasonal distribution of each of the sources of NOx can be clearly mapped from space, the team could provide independent topdown constraints on the individual strengths of NOx sources, and thus help resolve discrepancies in existing inventories. Jaegle's analysis of the satellite observations, presented at the recent Faraday Discussion on "Atmospheric Chemistry", shows that fuel combustion dominates emissions at northern mid-latitudes, while fires are a significant source in the Tropics. Additionally, she discovered a larger than expected role for soil emissions, especially over agricultural regions with heavy fertilizer use. Additional information is included in the original extended abstract.

Jaegle, Lyatt↗

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.↗

Comprehensive Measurement of Microbial Burden in Nutrient-Deprived Cleanrooms

Spacecraft surfaces that are destined to land on potential life-harboring celestial bodies are required to be rigorously cleaned and continuously monitored for spore bioburden as a proxy for spacecraft cleanliness. The NASA standard spore assay (NSA), used for spacecraft bioburden estimates, specifically measures spores that are cultivable, aerobic, resistant to heat shock, and grow at 30˚C in a nutrient-rich medium. Since the vast majority of microorganisms cannot be cultivated using the NSA assay, it is necessary to utilize state-of-the art molecular techniques to better understand the presence of all viable microorganisms, not just those measured with the NSA. In this study, the nutrient-deprived low biomass cleanrooms, where spacecraft are assembled, were used as a surrogate to spacecraft surfaces to measure the ratio of NSA spores in relation to the total viable microorganism population to compare with a 2006 space studies report that estimates that for every 1 spore there is approximately 50,000 viable organisms. Ninety-eight surface wipe samples were collected from the spacecraft assembly facility (SAF) cleanroom at the Jet Propulsion Laboratory (JPL) over a 6-month period. The samples were processed and analyzed using classical microbiology along with molecular assays. Traditional microbiology plating methods were used to determine the cultivable bacterial, fungal, and spore populations. Molecular assays were used to determine the total organisms (TO, dead and live) and the viable organisms (VO, live). The TO was measured using adenine triphosphate (ATP) and quantitative polymerase chain reaction (qPCR) assays. The VO was measured using internal ATP, propidium monoazide (PMA)-qPCR, and flow cytometry (after staining for viable microorganisms) assays. Based on the results, it was possible to establish a ratio between spore counts and VO for each viability assay. The ATP based spore to VO ratio ranged from 149 – 746 and the bacterial PMA-qPCR assay based ratio ranged from 314 – 1491 VO. The most conservative estimate came from FACS, which estimated the ratio to be 12,091 VO per 1 NSA spore. Since archaeal (<1%) and fungal (~2%) populations were negligible, the spore to VO ratios were based on bacterial population estimates. The most conservative ratio from this study can be used as a replacement for the SSB estimate on nutrient-deprived (oligotrophic) desiccated spacecraft surfaces, to estimate the VO from NSA measurements without utilizing state-of-the art molecular methods that are costly and require more biomass than is typically found of spacecraft surfaces.

Venkateswaran, Kasthuri↗

Response of soybean rhizosphere communities to human hygiene water addition as determined by community level physiological profiling (CLPP) and terminal restriction fragment length polymorphism (TRFLP) analysis

In this report, we describe an experiment conducted at Kennedy Space Center in the biomass production chamber (BPC) using soybean plants for purification and processing of human hygiene water. Specifically, we tested whether it was possible to detect changes in the root-associated bacterial assemblage of the plants and ultimately to identify the specific microorganism(s) which differed when plants were exposed to hygiene water and other hydroponic media. Plants were grown in hydroponics media corresponding to four different treatments: control (Hoagland's solution), artificial gray water (Hoagland's+surfactant), filtered gray water collected from human subjects on site, and unfiltered gray water. Differences in rhizosphere microbial populations in all experimental treatments were observed when compared to the control treatment using both community level physiological profiles (BIOLOG) and molecular fingerprinting of 16S rRNA genes by terminal restriction fragment length polymorphism analysis (TRFLP). Furthermore, screening of a clonal library of 16S rRNA genes by TRFLP yielded nearly full length SSU genes associated with the various treatments. Most 16S rRNA genes were affiliated with the Klebsiella, Pseudomonas, Variovorax, Burkholderia, Bordetella and Isosphaera groups. This molecular approach demonstrated the ability to rapidly detect and identify microorganisms unique to experimental treatments and provides a means to fingerprint microbial communities in the biosystems being developed at NASA for optimizing advanced life support operations.

NASA Discipline Life Support Systems↗

Developing Model Benchtop Systems for Microbial Experimental Evolution

Understanding how microbes impact an ecosystem has improved through advances of molecular and genetic tools, but creating complex systems that emulate natural biology goes beyond current technology. In fact, many chemical, biological, and metabolic pathways of even model organisms are still poorly characterized. Even then, standard laboratory techniques for testing microbial impact on environmental change can have many drawbacks; they are time-consuming, labor intensive, and are at risk of contamination. By having an automated process, many of these problems can be reduced or even eliminated. We are developing a benchtop system that can run for long periods of time without the need for human intervention, involve multiple environmental stressors at once, perform real-time adjustments of stressor exposure based on current state of the population, and minimize contamination risks. Our prototype device allows operators to generate an analogue of real world micro-scale ecosystems that can be used to model the effects of disruptive environmental change on microbial ecosystems. It comprises of electronics, mechatronics, and fluidics based systems to control, measure, and evaluate the before and after state of microbial cultures from exposure to environmental stressors. Currently, it uses four parallel growth chambers to perform tests on liquid cultures. To measure the population state, optical sensors (LED/photodiode) are used. Its primary selection pressure is UV-C radiation, a well-studied stressor known for its cell- and DNA-damaging effects and as a mutagen. Future work will involve improving the current growth chambers, as well as implementing additional sensors and environmental stressors into the system. Full integration of multiple culture testing will allow inter-culture comparisons. Besides the temperature and OD sensors, other types of sensors can be integrated such as conductivity, biomass, pH, and dissolved gasses such as CO and O. Additional environmental stressor systems like temperature (extreme heat or cold), metal toxicity, and other forms of radiation will increase the scale and testing range.

Developing↗

Bionutrients-1, On-Demand Production of Nutrients in Space

Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small “production pack” system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.

Hindupur, Aditya↗

BioNutrients-1: On-Demand Production of Nutrients in Space

Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small "production pack" system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.

Hindupur, Aditya↗

Biomarkers Detection With the Autonomous and Remotely Operated SOLID-LDChip Instrument in A Mars Drilling Simulating Campaign

One of the main goals in Mars exploration is to determine whether life has ever existed on the red planet. To achieve this goal, it is important to verify the performance, robustness, and maturity of instrumentation devoted to life detection, for example through field-testing in Mars analog environments.The Atacama Desert is considered one of the best terrestrial analogs of the surface of Mars due to abrupt temperature shifts, dryness, high UV radiation, and extremely low biomass. This, together with the lack of vegetation and the geochemistry of the regolith, make it an ideal scenario for testing instrumentation, performance, and concepts of operations. The Atacama Rover Astrobiology Drilling Studies (ARADS) is a NASA PSTAR project conceived for maturing and testing life-detection instrumentation in the Atacama Desert(1). The project is based on the K-REX2 rover equipped with a robotic 1-meter arm that delivers samples toon board instruments. This includes SOLID3.1 (Signs of Life Detector), designed as a life-detection instrument for finding complex organic molecules by means of the LDCHip (Life Detector Chip), an antibody microarray sensor(2). The instrument can extract organic compounds from soil, rocks, and sediments into a liquid solvent in the extraction cell (EC), allowing the search for hundreds of microbial molecular biomarkers at once (3). The robustness and reliability of SOLID were tested in the 2019 ARADS Mars drilling simulating campaign and confirmed with other analytical methods.

M Moreno-Paz↗

On the use of antibiotics to reduce rhizoplane microbial populations in root physiology and ecology investigations

No straightforward method exists for separating the proportion of ion exchange and respiration due to rhizoplane microbial organisms from that of root ion exchange and respiration. We examined several antibiotics that might be used for the temporary elimination of rhizoplane bacteria from hydroponically grown wheat roots (Triticum aestivum cv. Veery 10). Each antibiotic was tested for herbicidal activity and plate counts were used to enumerate bacteria and evaluate antibiotic kinetics. Only lactam antibiotics (penicillins and cephalosporins) did not reduce wheat growth rates. Aminoglycosides, the pyrimidine trimethoprim, colistin and rifampicin reduced growth rates substantially. Antibiotics acted slowly, with maximum reductions in rhizoplane bacteria occurring after more than 48 h of exposure. Combinations of nonphytotoxic antibiotics reduced platable rhizoplane bacteria by as much as 98%; however, this was generally a reduction from about 10(9) to 10(6) colony forming units per gram of dry root mass, so that many viable bacteria remained on root surfaces. We present evidence which suggests that insufficient bacterial biomass exists on root surfaces of nonstressed plants grown under well-aerated conditions to quantitatively interfere with root nitrogen absorption measurements.

NASA Discipline Life Support Systems↗

Tidal Marsh Outwelling of Dissolved Organic Matter and Resulting Temporal Variability in Coastal Water Optical and Biogeochemical Properties

Coastal wetlands are highly dynamic environments at the land-ocean interface where human activities, short-term physical forcings and intense episodic events result in high biological and chemical variability. Long being recognized as among the most productive ecosystems in the world, tidally-influenced coastal marshes are hot spots of biogeochemical transformation and exchange. High temporal resolution observations that we performed in several marsh-estuarine systems of the Chesapeake Bay revealed significant variability in water optical and biogeochemical characteristics at hourly time scales, associated with tidally-driven hydrology. Water in the tidal creek draining each marsh was sampled every hour during several semi-diurnal tidal cycles using ISCO automated samplers. Measurements showed that water leaving the marsh during ebbing tide was consistently enriched in dissolved organic carbon (DOC), frequently by more than a factor of two, compared to water entering the marsh during flooding tide. Estimates of DOC fluxes showed a net DOC export from the marsh to the estuary during seasons of both low and high biomass of marsh vegetation. Chlorophyll amounts were typically lower in the water draining the marsh, compared to that entering the marsh during flooding tide, suggesting that marshes act as transformers of particulate to dissolved organic matter. Moreover, detailed optical and compositional analyses demonstrated that marshes are important sources of optically and chemically distinctive, relatively complex, high molecular weight, aromatic-rich and highly colored dissolved organic compounds. Compared to adjacent estuarine waters, marsh-exported colored dissolved organic matter (CDOM) was characterized by considerably stronger absorption (more than a factor of three in some cases), larger DOC-specific absorption, lower exponential spectral slope, larger fluorescence signal, lower fluorescence per unit absorbance, and higher fluorescence at visible wavelengths. Observed patterns in water optical and biogeochemical variables were very consistent among different marsh systems and throughout the year, despite continued tidal exchange, implying rapid transformation of marsh DOM in the estuary through both photochemical and microbial processes. These findings illustrate the importance of tidal marsh ecosystems as sources, sinks and/or transformers of biologically important nutrients, carbon and colored dissolved organic compounds, and their influence on short-term biological, optical and biogeochemical variability in coastal waters.

Tzortziou, Maria↗

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↗

Preliminary Design of a Downstream Processing System for Protein Production in Space

Biomanufacturing is a promising technology to convert in situ resources into essential products including enzymes, therapeutics, biopolymers and other chemicals required to support deep-space missions that may not be easily supplied or produced by alternative means. In addition to the biomass production operations, vital down-stream steps including biomass harvesting/concentration, cell lysis, protein capture and purification are needed to produce an application-ready product. Commercially available terrestrial processes commonly require complex, heavy equipment and highly trained operators, which are not practical in deep space environments. In this work, we aim to identify approaches required to produce an intracellular, His-tagged recombinant enzyme using E. coli at 1 L production scales within the constraints of a deep-space mission as a model use-case scenario. Based on extensive literature review and commercially available products, we identified candidate technologies and products that could be integrated for deep space biomanufacturing. Different preliminary designs were then compared in terms of total system impacts on up-mass, processing time, and consumables required. Our analysis indicated that a biomass concentrator would significantly reduce the processing time and consumables required for the overall system without a large increase in the total mass. We also identified viable technologies for other steps such as cell lysis and protein purification. Predictions from our trade study will be validated in the laboratory by testing the most promising products with the results used to optimize the design. This research will help transfer technology that is well developed on Earth to a space-ready format to produce biological products from a wide variety of microorganisms that can support deep-space missions.

Biomanufacturing↗

Acronema sippewissettensis Gen. Nov. Sp. Nov., microbial mat bicosoecid (Bicosoecales = Bicosoecida)

A heterotrophic mastigote from the flat laminated Microcoleus-dominated intertidal microbial mat at the Sippewissett salt marsh, Cape Cod, Massachusetts, was isolated into monoprotist culture in the same anoxic medium that led to spirochete and other anaerobic bacterial enrichments. The protist grew vigorously and was transferred indefinitely in oxic marine medium. Videomicroscopy as well as scanning and transmission electron microscopy were used to document its features. The swimming and perching behavior, nutritional mode (bactivory) and morphology including ultra-structure identify it as an aloricate bicosoecid. The presence of heteromorphic acronematic undulipodia, bilateral bipartite tubular mastigonemes, absence of a cytostome, absence of extrusomes, and presence of "Dauerstadien" (duration stages) distinguish this from other Cafeteriaceae bicosoecids. Cell division involves a closed intranuclear spindle. The unspecialized bicosoecid morphology and behavior juxtaposed with oomycete-like vesicles and mastigonemes suggest that this protist may be an extant descendant of a common ancestor of bicosoecids and other stramenopiles (e.g. labyrinthulids, thraustochytrids and oomycetes). A new genus and species, Acronema sippewissettensis, are proposed.

Non-NASA Center↗

Quantitative determination of microbial activity and community nutritional status in estuarine sediments: evidence for a disturbance artifact

In estuarine sediments with a high degree of vertical heterogeneity in reduced substrate and terminal electron acceptor concentrations, the method of exposure of the microbiota to labeled substrates can introduce a "disturbance artifact" into measures of metabolic activity. The detection of this artifact is based on quantitative measurement of the relative rates of incorporation of [14C]acetate into phospholipid fatty acids (PLFA) and endogenous storage lipid, poly-beta-hydroxyalkanoate (PHA). Previous studies have shown that PLFA synthesis measures cellular growth and that PHA synthesis measures carbon accumulation (unbalanced growth). The "disturbance artifact" of exposure to [14C]acetate was demonstrated by comparing injection of a core with the usual or pore-water replacement or slurry techniques. Only injection of labeled substrate allowed detection of preassay disturbance of the sediment with a garden rake. The raking increased PLFA synthesis with little effect to differences in concentration or distribution of [14C]acetate in the 10-min incubation. Bioturbation induced by sand dollar feeding in estuarine sediment could be detected in an increased PLFA/PHA ratio which was due to decreased PHA synthesis if the addition of labeled substrate was by the injection technique. Addition of labeled precursors to sediment by slurry or pore-water replacement induces greater disturbance artifacts than injection techniques.

Phospholipids/analysis/biosynthesis↗

Model system for studies of microbial dynamics at exuding surfaces such as the rhizosphere

An autoclavable all-glass system for studying microbial dynamics at permeable surfaces is described. Standard hydrophobic or hydrophilic membranes (46-mm diameter) of various pore sizes were supported on a glass frit through which nutrient solutions were pumped by a peristaltic pump. The pump provided a precisely controlled flow at speeds of 0.5 to 500 ml of defined or natural cell exudates per h, which passed through the membrane into a receiving vessel. The construction allowed a choice of membranes, which could be modified. The system was tested with a bacterium, isolated from rape plant roots (Brassica napus L.), that was inoculated on a hydrophilic membrane filter and allowed to develop into a biofilm. A defined medium with a composition resembling that of natural rape root exudate was pumped through the membrane at 0.5 ml/h. Scanning electron microscopic examinations indicated that the inoculum formed microcolonies embedded in exopolymers evenly distributed over the membrane surface. The lipid composition and content of poly-beta-hydroxybutyrate in free-living and adhered cells were determined by gas chromatography. The bacterial consumption of amino acids in the exudate was also studied.

NASA Discipline Number 61-10↗