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NASA NTRS2018

Intra-EVA Space-to-Ground Interactions when Conducting Scientific Fieldwork Under Simulated Mars Mission Constraints

The Biologic Analog Science Associated with Lava Terrains (BASALT) project is a four-year program dedicated to iteratively designing, implementing, and evaluating concepts of operations (ConOps) and supporting capabilities to enable and enhance scientific exploration for future human Mars missions. The BASALT project has incorporated three field deployments during which real (non-simulated) biological and geochemical field science have been conducted at two high-fidelity Mars analog locations under simulated Mars mission conditions, including communication delays and data transmission limitations. BASALT's primary Science objective has been to extract basaltic samples for the purpose of investigating how microbial communities and habitability correlate with the physical and geochemical characteristics of chemically altered basalt environments. Field sites include the active East Rift Zone on the Big Island of Hawai'i, reminiscent of early Mars when basaltic volcanism and interaction with water were widespread, and the dormant eastern Snake River Plain in Idaho, similar to present-day Mars where basaltic volcanism is rare and most evidence for volcano-driven hydrothermal activity is relict. BASALT's primary Science Operations objective has been to investigate exploration ConOps and capabilities that facilitate scientific return during human-robotic exploration under Mars mission constraints. Each field deployment has consisted of ten extravehicular activities (EVAs) on the volcanic flows in which crews of two extravehicular and two intravehicular crewmembers conducted the field science while communicating across time delay and under bandwidth constraints with an Earth-based Mission Support Center (MSC) comprised of expert scientists and operators. Communication latencies of 5 and 15 min one-way light time and low (0.512 Mb/s uplink, 1.54 Mb/s downlink) and high (5.0 Mb/s uplink, 10.0 Mb/s downlink) bandwidth conditions were evaluated. EVA crewmembers communicated with the MSC via voice and text messaging. They also provided scientific instrument data, still imagery, video streams from chest-mounted cameras, GPS location tracking information. The MSC monitored and reviewed incoming data from the field across delay and provided recommendations for pre-sampling and sampling tasks based on their collective expertise. The scientists used dynamic priority ranking lists, referred to as dynamic leaderboards, to track and rank candidate samples relative to one another and against the science objectives for the current EVA and the overall mission. Updates to the dynamic leaderboards throughout the EVA were relayed regularly to the IV crewmembers. The use of these leaderboards enabled the crew to track the dynamic nature of the MSC recommendations and helped minimize crew idle time (defined as time spent waiting for input from Earth during which no other productive tasks are being performed). EVA timelines were strategically designed to enable continuous (delayed) feedback from an Earth-based Science Team while simultaneously minimizing crew idle time. Such timelines are operationally advantageous, reducing transport costs by eliminating the need for crews to return to the same locations on multiple EVAs while still providing opportunities for recommendations from science experts on Earth, and scientifically advantageous by minimizing the potential for cross-contamination across sites. This paper will highlight the space-to-ground interaction results from the three BASALT field deployments, including planned versus actual EVA timeline data, ground assimilation times (defined as the amount of time available to the MSC to provide input to the crew), and idle time. Furthermore, we describe how these results vary under the different communication latency and bandwidth conditions. Together, these data will provide a basis for guiding and prioritizing capability development for future human exploration missions.

Beaton, Kara H.↗
NASA NTRS2017

Space Plants for Astronaut Consumption

Growing plants in space will be an essential part of sustaining astronauts during long-range missions. During the summer of 2017, three female NASA interns, have been engaged in research relevant to food production in space, and will present their projects to an all female program known as Girls in STEM camp. Ayla Grandpre, a senior from Rocky Mountain College, has performed data mining and analysis of crop growth results gathered through Fairchild Botanical Gardens program, Growing Beyond Earth. Ninety plants were downselected to three for testing in controlled environment chambers at KSC. Ayla has also managed an experiment testing a modified hydroponics known as PONDS, to grow mizuna mustard greens and red robin cherry tomatoes. Emma Boehm, a senior from the University of Minnesota, has investigated methods to sterilize seeds and analyzed the most common microbial communities on seed surfaces. She has tested a bleach fuming method and an ethanol treatment. Emma has also tested Tokyo bekana Chinese cabbage seeds from four commercial seed vendors to identity differences in germination and growth variability. Lastly, Payton Barnwell, a junior from Florida Polytechnic University has shown that light recipes provided by LEDs can alter the growth and nutrition of 'Outredgeous' lettuce, Chinese cabbage, and Mizuna. The results of her light quality experiments will provide light recipe recommendations for space crops that grown in the Advanced Plant Habitat currently aboard the International Space Station.

VEGGIE↗
NASA NTRS2018

A Prototype Tool for Assessing Forward Contamination from Space Suits

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

Rucker, M.↗
NASA NTRS2018

Space-to-Ground Interactions While Conducting Scientific Fieldwork Under Mars Mission Constraints

The Biologic Analog Science Associated with Lava Terrains (BASALT) project is a 4-year program dedicated to iteratively designing, implementing, and evaluating concepts of operations (ConOps) and supporting capabilities to enable and enhance scientific exploration for future human Mars missions. BASALT incorporates three field deployments during which real (non-simulated) biological and geochemical field science is conducted at two high-fidelity Mars analog locations under simulated Mars mission conditions, including communication de-lays and data transmission limitations. BASALT’s primary science objective is to investigate how the redox conditions of altered basaltic environments affect the development of microbial communities in these Mars-relevant settings. Field sites include the active East Rift Zone on the Big Island of Hawaii, reminiscent of early Mars when basaltic volcanism and interaction with water were widespread, and the dormant eastern Snake River Plain in Idaho, similar to present-day Mars where basaltic volcanism is rare and most evidence for volcano-driven hydrothermal activity is relict. BASALT’s primary science operations objective is to investigate exploration ConOps and capabilities that facilitate scientific return during human-robotic exploration under Mars mission constraints. Each field deployment consists of ten extravehicular activities (EVAs) on the volcanic flows in which two extravehicular and two intravehicular (IV) crew-members conduct the science while communicating across time delay and under bandwidth constraints with an Earth-based Mission Support Center (MSC) comprised of expert scientists and operators. Communication latencies of 5 and 15-minute one-way light time and low (0.512 Mb/s uplink, 1.54 Mb/s down-link) and high (5.0 Mb/s uplink, 10.0 Mb/s downlink) bandwidth conditions are being evaluated. EVA crewmembers communicate with the MSC via voice and text messaging and provide scientific instrument data, still imagery, video streams, and GPS tracking information. The MSC reviews this data across delay and provides recommendations for presampling and sampling tasks. The scientists used dynamic leaderboards (priority rank-ing lists), to track and rank candidate samples relative to one another and against the science objectives for the current EVA and the overall mission. Updates to the dynamic leaderboards are relayed regularly to the IV crewmembers to provide scientific feedback from Earth and to help minimize crew idle time (time spent waiting for Earth input during which no productive tasks are performed). EVA timelines are strategically designed to enable continuous (delayed) feedback from an Earth-based science team while simultaneously minimizing crew idle time. Such timelines are operationally advantageous, reducing transport costs by eliminating the need for crews to return to the same locations on multiple EVAs while still providing opportunities for recommendations from science experts on Earth, and scientifically advantageous by minimizing the potential for cross-contamination across sites. This paper will highlight the space-to-ground interaction results from the three BASALT field deployments, including planned versus actual EVA time-line data, ground assimilation times (the amount of time available to the MSC to provide input to the crew), and idle time. Furthermore, we describe how these results vary under the different communication latency and bandwidth conditions. Together, these data will provide a basis for guiding and prioritizing capability development for future human exploration missions.

Beaton, Kara H.↗
NASA NTRS2019

Using the NASA GeneLab Data System to Study the Metagenomes of Spaceships and Their Occupants

With humans pushing to live further off Earth for longer periods of time, it is increasingly important to understand the changes that occur in biological systems during spaceflight whether these be astronauts, their microbial commensals, or their plant-based life support systems. In a three-part presentation, we discuss GeneLab and recent discoveries regarding the microbiota of spacecrafts and space-flown animals. Part 1: GeneLab: Open Science for Life in Space, Jonathan Galazka, NASA Ames Research Center To accelerate the pace of discovery from precious spaceflight biological experiments, NASA as develop the GeneLab data system (genelab.nasa.gov), which allows unfettered access to omics data from spaceflight and spaceflight relevant experiments. GeneLab houses metagenomic datasets from spacecraft and relevant spacecraft models. Users can download this data and associated metadata to make new discoveries about how microbial communities may change and adapt to spaceflight.

Galazka, Jonathan M.↗
NASA NTRSDate not supplied

Seed surface sanitization and persistence of E.coli through different tissues of ‘Red Robin’ Tomato (Solanum lycopersicum cv. Red Robin)

Seed surface sanitization via chemical processes removes/reduces microbes from the external surfaces of the seed and thereby could have an impact on the plants’ health or productivity. To determine the impact of seed surface sanitization on the plants’ microbiome, sanitized and unsanitized seeds from ‘Red Robin’ Tomato (Solanumlycopersicum cv. Red Robin) were exposed to Escherichia coli (E. coli) and grown in a controlled environment growth chamber simulating environmental conditions aboard the International Space Station (ISS). Plants were harvested at four intervals, days 11, 33,42 and 76 post-germination. Changes in the microbial communities of leaf, stem, root, and fruit because of E. coli exposure and the persistence of E. coli itself were investigated using aerobic plate count (APC), qPCR and 16S rRNA sequencing. It was determined that E. coli persisted for longer periods of time in plants from sanitized versus unsanitized seeds and was identified in root tissue more frequently than in leaf or stem tissue. E. coli was not detected in fruits raised from either sanitized or unsanitized seeds. The 16S rRNA sequencing showed dynamic changes in the abundance of members of the phylum Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes in all tissue types studied. We observed minimal or no changes in the alpha diversity of leaf stem and fruit tissue with time, or between sanitized and unsanitized seeds. Roots showed significant differences in alpha diversity with time and seed sanitization status. Beta-diversity showed that time had more of an influence on all samples versus the E. coli treatment. Members of phyla Proteobacteria and Bacteroidetes were found to be differentially abundant across leaf, stem and root tissue. Our results indicated that the seed surface sanitization, although a requirement for sending seeds to space, might influence the developing microbiome. This research was funded by NASA’s Space Life and Physical Sciences Research and Applications.

Anirudha R Dixit↗
NASA NTRS2020

The Atacama Desert: A Window into Late Mars Surface Habitability?

Decades of robotic exploration have revealed that Mars was capable of sustaining life in the Noachian and Hesperian periods (> 3 billion years ago). But habitability conditions progressively deteriorated thereafter, and a lasting trend of climate change has made the surface of the planet likely uninhabitable at present times. One of the outstanding enigmas in Mars exploration is when was the last time that the surface of the planet could have sustained life. In this chapter we address this question through comparisons with one of the oldest and driest deserts on Earth, the Atacama Desert in northern Chile. We base our analysis on a latitudinal rainfall gradient that allows to investigate how microbial communities respond to increasing dryness. We posit that the ecological successions observed along the Atacama rainfall gradient represent a possible model for the adaptation of an early martian biosphere to increasing dryness throughout the planet’s geologic history, offering clues to the last surface environments that could have sustained life, and the timing of the last surface habitable period. These might in turn be the best locations to search for evidence of life.

Atacama Desert↗
NASA NTRSDate not supplied

Hydrogen Peroxide as a Method for Bioburden Reduction in Facilities with Strict Materials Requirements.

The cleanrooms used to curate NASA’s Astromaterials samples are carefully monitored for particulate and inorganic contamination. The clean labs also have a very limited set of acceptable materials and cleaning agents to further minimize the potential for contamination. Labs are cleaned primarily with isopropyl alcohol. Astromaterials samples are handled with tools made of stainless steel (304 or 316), Teflon, or aluminum alloy (6061). Although our current collections are not particularly susceptible to biological alteration or organic contamination, this will not be the case for new collections from the OSIRIS-REx mission, Hayabusa2, and from Mars Sample Return. Therefore, it is necessary to develop and test methods to reduce the bioburden in astromaterials cleanrooms without introducing unwanted contaminants. We will report on the results of three case studies where 7.5 wt% hydrogen peroxide was prepared from a stock solution of ultrapure 30 wt% hydrogen peroxide (JT Baker) using curation-grade ultrapure water. We followed CDC (Center for Disease Control) guidelines for using hydrogen peroxide as a high level disinfectant. This solution was used to clean a glovebox prior to processing Apollo samples, as well as surfaces in the Antarctic meteorite processing lab and Stardust lab after facilities monitoring indicated an unwanted increase in bioburden. In all three instances, the culturable bioburden was significantly reduced after a 30 min. exposure to the 7.5% hydrogen peroxide solution without a corresponding increase in inorganic or organic contamination. We observed 77 to 100% reductions in the bioburden recovery rate. In one case study, we also performed amplicon DNA sequencing on samples collected from the surfaces before and after cleaning. We observed a significant change in microbial community composition after peroxide cleaning. These results suggest that routine cleaning with hydrogen peroxide could be an effective way to control bioburden in astromaterials cleanrooms and other facilities with strict contamination control requirements.

A B Regberg↗
NASA NTRSDate not supplied

Understanding amyloids to prevent biofilm formation in space

There is a pressing need to search for novel approaches to combat biofilm formation, both in space and in medical applications. Many proteins have the ability to form ordered aggregates called amyloids. Amyloids are known to be an important part of biofilms. The use of anti-amyloid drugs is a novel venue for the development of antimicrobial agents. The ultrastructure of the amyloid aggregate shows a high packing of proteins, the second-order structure of which is dominated by β-sheets. The ability to form an amyloid aggregate is especially typical for proteins containing domains (protein fragments) with sufficient lability to arrange themselves in a tight β-sheet structure. Bioinformatics tools allow the prediction of such behavior of proteins in genomic data. We use GeneLab data of microbial populations identified aboard the International Space Station and other spacecraft to look for bacterial species that utilize amyloid aggregation in biofilm formation. We use a combined bioinformatic approach with a relatively high throughput molecular biology assay and biophysical assays to evaluate the anti-amyloid anti-biofilm approach. The significance of the research extends from understanding basic microbial community responses to spaceflight, to biofouling of the built environments in space as well as the long-term health of astronauts. Bioinformatics shows that onboard the ISS, bacterial species produce far more amyloid and prion proteins than are currently verified, hence their role in bacterial ecosystems is largely unknown. As we propose there is a link between amyloid formation in space and biofilm production, this research should lead to new paths for biofilm remediation in space.

Tomasz Zajkowski↗
NASA NTRSDate not supplied

Hydrogen Peroxide as a Method for Bioburden Reduction in Facilities with Strict Materials Requirements

The cleanrooms used to curate NASA’s Astromaterials samples are carefully monitored for particulate and inorganic contamination. The clean labs also have a very limited set of acceptable materials and cleaning agents to further minimize the potential for contamination. Labs are cleaned primarily with isopropyl alcohol. Astromaterials samples are handled with tools made of stainless steel (304 or 316), Teflon, or aluminum alloy (6061). Although our current collections are not particularly susceptible to biological alteration or organic contamination, this will not be the case for new collections from the OSIRIS-REx mission, Hayabusa2, and from Mars Sample Return. Therefore, it is necessary to develop and test methods to reduce the bioburden in astromaterials cleanrooms without introducing unwanted contaminants. We will report on the results of three case studies where 7.5 wt% hydrogen peroxide was prepared from a stock solution of ultrapure 30 wt% hydrogen peroxide (JT Baker) using curation-grade ultrapure water. We followed CDC (Center for Disease Control) guidelines for using hydrogen peroxide as a high level disinfectant. This solution was used to clean a glovebox prior to processing Apollo samples, as well as surfaces in the Antarctic meteorite processing lab and Stardust lab after facilities monitoring indicated an unwanted increase in bioburden. In all three instances, the culturable bioburden was significantly reduced after a 30 min. exposure to the 7.5% hydrogen peroxide solution without a corresponding increase in inorganic or organic contamination. We observed 77 to 100% reductions in the bioburden recovery rate. In one case study, we also performed amplicon DNA sequencing on samples collected from the surfaces before and after cleaning. We observed a significant change in microbial community composition after peroxide cleaning. These results suggest that routine cleaning with hydrogen peroxide could be an effective way to control bioburden in astromaterials cleanrooms and other facilities with strict contamination control requirements.

A B Regberg↗
NASA NTRSDate not supplied

Effects of Spaceflight Relevant Carbon Dioxide Levels on Pathogenesis Related Microbial Characteristics

BACKGROUND Multiple stressors in the spaceflight environment have the potential to alter microbial pathogenesis and virulence characteristics, including microgravity, radiation, and hostile/confined conditions. This combination of factors may lead to a stacking of risks, potentially creating unexpected increase in the risk of infectious disease. One understudied variable is the higher CO2 atmospheric concentration. The current Spaceflight Maximum Allowance Concentration (SMAC) for 24-hour average CO2 is 0.4% (3 mm Hg). These elevated ambient CO2 levels aboard the ISS could potentially influence the diversity and phenotypic responses of the resident microbial communities from both the spacecraft environment (air, surface, water) and crewmembers (gut, nasal, skin microbiomes). METHODS For these studies, we will evaluate the response of Staphylococcus aureus, Streptococcus pneumoniae and Enterobacter aerogenes, which are opportunistic pathogens that are of medical significance and have been or are likely to be found aboard spacecraft. These microorganisms will be cultured in both spaceflight analog and control conditions at CO2levels representing terrestrial atmospheric concentration of 0.04% (0.3 mmHg), ISS elevated concentration of 0.4% (3 mm Hg), and an unexpected elevated concentration of 1 % (7.6 mmHg) to represent a worst case scenario. DISCUSSION Overwhelming evidence from studies performed in food microbiology, marine biology and terrestrial environmental biology suggest altered gene expression, selective bacterial inhibition, increased growth and diversity, and increased antibiotic resistance of bacterial communities individually and in biofilm formation when exposed to increased levels of CO2. While many of these findings are based on levels of CO2 higher than would be found during spaceflight exploration missions, the potential for subtle changes in CO2 levels to exacerbate infectious disease risks warrants an evaluation of pathogen responses to spaceflight CO2 conditions. Our study will provide answers to spaceflight relevant atmospheric conditions, as described in the Risk of Adverse Health Effects Due to Host-Microorganism Interaction (GapMicro 103).

A A Medina-Colorado↗
NASA NTRSDate not supplied

American Society of Plant Biologists Plenary Symposium Overview Presentation

NASA’s Artemis program in collaboration with international space agencies, and various commercial partners, marks the return of humankind to the Moon. Unlike the Apollo lunar missions from fifty years ago, Artemis will set the stage for the continuous presence of humans on the Moon and pave a path for the journey to Mars and beyond. Plants will be an essential component of habitation systems that will enable humans to thrive on both the lunar and Mars surface, as well as space transit vehicles because of their nutritional and psychological benefits. Furthermore, plants will be valued in environmental control and life support systems because they generate oxygen, remove carbon dioxide, and recycle water and waste. However, an in-depth understanding of how plants and their associated microbial communities adapt to the harsh and confined environment of deep space is essential before they can be effectively used for long-duration space missions. This plenary symposium will address challenges and present potential solutions to growing plants in space. The four speakers will cover a broad range of topics, including mechanistic studies on fern resurgence after asteroid-triggered mass extinction events, artificial photosynthesis for space agriculture, and how the plant microbiome and an understanding of plant-pathogen dynamics in microgravity can guide crop production strategies in space.

Plant Space Biology↗
NASA NTRS1985

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↗
NASA NTRS2011

Microbial Diversity Analysis of the Bacterial and Archaeal Population in Present Day Stromatolites

Stromatolites are layered sedimentary structures resulting from microbial mat communities that remove carbon dioxide from their environment and biomineralize it as calcium carbonate. Although prevalent in the fossil record, stromatolites are rare in the modem world and are only found in a few locations including Highbome Cay in the Bahamas. The stromatolites found at this shallow marine site are analogs to ancient microbial mat ecosystems abundant in the Precambrian period on ancient Earth. To understand how stromatolites form and develop, it is important to identify what microorganisms are present in these mats, and how these microbes contribute to geological structure. These results will provide insight into the molecular and geochemical processes of microbial communities that prevailed on ancient Earth. Since stromatolites are formed by lithifying microbial mats that are able to mineralize calcium carbonate, understanding the biological mechanisms involved may lead to the development of carbon sequestration technologies that will be applicable in human spaceflight, as well as improve our understanding of global climate and its sustainability. The objective of my project was to analyze the archaeal and bacterial dIversity in stromatolites from Highborn Cay in the Bahamas. The first step in studying the molecular processes that the microorganisms carry out is to ascertain the microbial complexity within the mats, which includes identifying and estimating the numbers of different microbes that comprise these mats.

Ortega, Maya C.↗
NASA NTRS1995

Thermal Springs and the Search for Past Life on Mars

Ancient thermal spring sites have several features which make them significant targets in a search for past life. Chemical (including redox) reactions in hydrothermal systems possibly played a role in the origin of life on Earth and elsewhere. Spring waters frequently contain reduced species (sulfur compounds, Fe(sup +2), etc.) which can provide chemical energy for organic synthesis. Relatively cool hydrothermal systems can sustain abundant microbial life (on Earth, at temperatures greater than 110 C). A spring site on Mars perhaps might even have maintained liquid water for periods sufficiently long to sustain surface-dwelling biota had they existed. On Earth, a variety of microbial mat communities can be sampled along the wide range of temperatures surrounding the spring, thus offering an opportunity to sample a broad biological diversity. Thermal spring waters frequently deposit minerals (carbonates, silica, etc.) which can entomb and preserve both fluid inclusions and microbial communities. These deposits can be highly fossiliferous and preserve biological inclusions for geologically long periods of time. Such deposits can cover several square km on Earth, and their distinctive mineralogy (e.g., silica- and/or carbonate-rich) can contrast sharply with that of the surrounding region. As with Martian volcanoes, Martian thermal spring complexes and their deposits might typically be much larger than their counterparts on Earth. Thus Martian spring deposits are perhaps readily detectable and even accessible. Elysium Planitia is an example of a promising region where hydrothermal activity very likely remobilized ground ice and sustained springs.

DesMarais, D. J.↗
NASA NTRS2012

Microbial Lipid and C Isotopic Biosignatures of a Unique Community at Grand Prismatic Spring, Yellowstone National Park

The microbial communities found in modern hot springs are considered analogs to ones that may have existed in hydrothermal systems on the early Earth and possibly Mars. Our goal was to characterize the microbial biosignatures and to assess the preservation of organic matter in the silica-depositing Grand Prismatic Spring in Yellowstone National Park. This study combines 16S rRNA surveys, lipid biomarkers, and C isotopes to query, "Who's there and what are they doing?" On the edge of the approximately 90 m diameter blue vent pool (56.1 C, pH 8.5), a floating green streamer community grew over a benthic pink community. The membrane lipids in the green streamers and pink mat were composed of unusual ester-linked fatty acids, indicating the presence of novel bacterial groups. In particular, we discovered a series of 2-methyl and 2,X-dimethyl phospholipid fatty acids (C18-22). We are attempting to use the 16S rRNA surveys to link these compounds to source organisms. Wax esters, biomarkers for Chloroflexi, were present in both communities, but displayed different profiles. A higher proportion of branched wax esters were found in the green streamers, and were associated with a relatively high concentration of long-chain di- and trienes (C29-31). This suggests that Chloroflexus primarily grew in the green streamers, while a pink mat of Roseiflexus grew on the sinter substrate underneath. Cyanobacterial alkanes were found in the green streamers (n-C17, 7-, 6- and 5-monomethyl-C17, 7,11-dimethyl-C17, n-C19, n-C19:1). We also detected a series of monoalkylglycerylethers and geologically relevant hopanoids in both communities. Carbon isotope analyses indicated that Chloroflexus was growing photoheterotrophically using cyanobacterial photosynthate. Roseiflexus also traditionally grows photoheterotrophically, but the C isotopic signatures of the lipids in the pink mat were approximately 10 %0 lighter than the cyanobacterial and Chloroflexus lipids, indicating a potentially novel metabolic mechanism or prior secondary reworking of substrates before reaching Roseiflexus. This arrangement of Synechococcus-Chloroflexus green streamers floating over a benthic pink community of Roseiflexus is different from the classical laminated Synechococcus-Chloroflexi mats at Octopus Hot Spring.

Jahnke, Linda↗
NASA NTRS1998

A natural view of microbial biodiversity within hot spring cyanobacterial mat communities

This review summarizes a decade of research in which we have used molecular methods, in conjunction with more traditional approaches, to study hot spring cyanobacterial mats as models for understanding principles of microbial community ecology. Molecular methods reveal that the composition of these communities is grossly oversimplified by microscopic and cultivation methods. For example, none of 31 unique 16S rRNA sequences detected in the Octopus Spring mat, Yellowstone National Park, matches that of any prokaryote previously cultivated from geothermal systems; 11 are contributed by genetically diverse cyanobacteria, even though a single cyanobacterial species was suspected based on morphologic and culture analysis. By studying the basis for the incongruity between culture and molecular samplings of community composition, we are beginning to cultivate isolates whose 16S rRNA sequences are readily detected. By placing the genetic diversity detected in context with the well-defined natural environmental gradients typical of hot spring mat systems, the relationship between gene and species diversity is clarified and ecological patterns of species occurrence emerge. By combining these ecological patterns with the evolutionary patterns inherently revealed by phylogenetic analysis of gene sequence data, we find that it may be possible to understand microbial biodiversity within these systems by using principles similar to those developed by evolutionary ecologists to understand biodiversity of larger species. We hope that such an approach guides microbial ecologists to a more realistic and predictive understanding of microbial species occurrence and responsiveness in both natural and disturbed habitats.

Review↗
NASA NTRS1991

A model for diurnal patterns of carbon fixation in a Precambrian microbial mat based on a modern analog

Microbial mat communities are one of the first and most prevalent biological communities known from the Precambrian fossil record. These fossil mat communities are found as laminated sedimentary rock structures called stromatolites. Using a modern microbial mat as an analog for Precambrian stromatolites, a study of carbon fixation during a diurnal cycle under ambient conditions was undertaken. The rate of carbon fixation depends primarily on the availability of light (consistent with photosynthetic carbon fixation) and inorganic carbon, and not nitrogen or phosphorus. Atmospheric PCO2 is thought to have decreased from 10 bars at 4 Ga (10(9) years before present) to approximately 10(-4) bars today, implying a change in the availability of inorganic carbon for carbon fixation. Experimental manipulation of levels of inorganic carbon to levels that may have been available to Precambrian mat communities resulted in increased levels of carbon fixation during daylight hours. Combining these data with models of daylength during the Precambrian, models are derived for diurnal patterns of photosynthetic carbon fixation in a Precambrian microbial mat community. The models suggest that, even in the face of shorter daylengths during the Precambrian, total daily carbon fixation has been declining over geological time, with most of the decrease having occurred during the Precambrian.

Review↗