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DOE OSTI2022

A legacy of fire emerges from multiple disturbances to most shape microbial and nitrogen dynamics in a deciduous forest

Healthy forests are vital components of terrestrial ecosystems for their raw materials, high biodiversity, cycling of nutrients, and potential to sequester carbon. However, these ecosystems are sensitive to disturbances, and anthropogenic activities pose a serious threat to forest ecosystems globally. For example, human activities have dramatically altered multiple historical disturbance regimes in forests, including suppressing fire, increasing the density of large herbivores, and reducing the size of canopy gaps, among other disturbances. Such disturbances can have dramatic impacts on microbially-mediated forest soil functions, but more research is needed to determine the collective impacts of these disturbances. In this study, we investigated the interactive effects of disturbances, namely the legacies of fire, large herbivore densities, and canopy gap creation, in a deciduous forest soil. We determined that forest floor and mineral soil carbon and nitrogen pools were shaped by multiple disturbances, but fire was more influential than the other disturbances. The abundance of several functionally-relevant microbial taxa were significantly impacted by fire, and the effect was more pronounced in the mineral soil than in the forest floor. Together, these findings demonstrate that multiple disturbances, especially a legacy of fire, exerts long-term control over soil carbon, nitrogen and microbial dynamics in a deciduous forest system.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

Using Methyl Bromide for Interspecies Cell–Cell Signaling and As a Reporter in a Model Soil Consortium

Synthetic microbial communities are emerging as model systems for studying consortia-scale phenotypes. To establish synthetic biology tools for studying these communities in hard-to-image environmental materials, we evaluated whether a single member of a model soil consortium (MSC) can be programmed to report on gene expression without requiring matrix disruption. For these studies, we targeted a five member MSC that includes Dyadobacter fermentans, Ensifer adhaerens, Rhodococcus sp003130705, Streptomyces sp001905665, and Variovorax beijingensis. By coupling the expression of a methyl halide transferase to a constitutive promoter, we show that Variovorax beijingensis can be programmed to synthesize methyl halides that accumulate in the soil headspace at levels that are ≥24-fold higher than all other MSC members across a range of environmentally-relevant hydration conditions. We find that methyl halide production can report on a MSC promoter that is activated by changes in water potential, and we demonstrate that a synthetic gas signal can be read out directly using gas chromatography and indirectly using a soil-derived Methylorubrum that is programmed to produce a visual output in response to methyl halides. Finally, these tools will be useful for future studies that investigate how MSC respond to dynamic hydration conditions, such as drought and flood events induced by climate change, which can alter soil water potential and induce the release of stored carbon.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Diverse MarR bacterial regulators of auxin catabolism in the plant microbiome

Abstract Chemical signalling in the plant microbiome can have drastic effects on microbial community structure, and on host growth and development. Previously, we demonstrated that the auxin metabolic signal interference performed by the bacterial genus Variovorax via an auxin degradation locus was essential for maintaining stereotypic root development in an ecologically relevant bacterial synthetic community. Here, we dissect the Variovorax auxin degradation locus to define the genes iadDE as necessary and sufficient for indole-3-acetic acid (IAA) degradation and signal interference. We determine the crystal structures and binding properties of the operon’s MarR-family repressor with IAA and other auxins. Auxin degradation operons were identified across the bacterial tree of life and we define two distinct types on the basis of gene content and metabolic products: iac -like and iad -like. The structures of MarRs from representatives of each auxin degradation operon type establish that each has distinct IAA-binding pockets. Comparison of representative IAA-degrading strains from diverse bacterial genera colonizing Arabidopsis plants show that while all degrade IAA, only strains containing iad -like auxin-degrading operons interfere with auxin signalling in a complex synthetic community context. This suggests that iad -like operon-containing bacterial strains, including Variovorax species, play a key ecological role in modulating auxins in the plant microbiome.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Low-abundance populations distinguish microbiome performance in plant cell wall deconstruction

Abstract Background Plant cell walls are interwoven structures recalcitrant to degradation. Native and adapted microbiomes can be particularly effective at plant cell wall deconstruction. Although most understanding of biological cell wall deconstruction has been obtained from isolates, cultivated microbiomes that break down cell walls have emerged as new sources for biotechnologically relevant microbes and enzymes. These microbiomes provide a unique resource to identify key interacting functional microbial groups and to guide the design of specialized synthetic microbial communities. Results To establish a system assessing comparative microbiome performance, parallel microbiomes were cultivated on sorghum ( Sorghum bicolor L. Moench) from compost inocula. Biomass loss and biochemical assays indicated that these microbiomes diverged in their ability to deconstruct biomass. Network reconstructions from gene expression dynamics identified key groups and potential interactions within the adapted sorghum-degrading communities, including Actinotalea , Filomicrobium , and Gemmatimonadetes populations. Functional analysis demonstrated that the microbiomes proceeded through successive stages that are linked to enzymes that deconstruct plant cell wall polymers. The combination of network and functional analysis highlighted the importance of cellulose-degrading Actinobacteria in differentiating the performance of these microbiomes. Conclusions The two-tier cultivation of compost-derived microbiomes on sorghum led to the establishment of microbiomes for which community structure and performance could be assessed. The work reinforces the observation that subtle differences in community composition and the genomic content of strains may lead to significant differences in community performance.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

CFD Simulation of Aerobic Gas Fermentation to Enable Commercial Conversion of CO 2 into Aquaculture and Animal Feed: Cooperative Research and Development (Final Report)

NovoNutrients’ fermentation technology uses energy from hydrogen to transform industrial CO2 emissions into premium animal feed ingredients and other valuable products. A single NovoNutrients’ commercial manufacturing plant will capture and convert over 200,000 tons/yr of CO2 into over 100,000 tons/yr of high-protein feed. Key to the rapid and widespread deployment of the technology is maximization of its productivity and energy efficiency. Robust, physically based computational models of the technology will significantly increase productivity and efficiency, accelerating NovoNutrients’ technology to manufacturing scale. NREL has unique capabilities for creating and running such computational models. NREL's existing aerobic bioreaction computational fluid dynamics (CFD) models will be adapted to NovoNutrients’ gas fermentation (CO2, H2, O2) technology. The multiphysics CFD simulations require thousands of high-performance computing (HPC) node hours to simulate the complex geometries and contents of NovoNutrients’ industrial bioreactors. The experimentally validated CFD models were used to identify optimally efficient and productive bioreactor designs and operating conditions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2022

Versatile soil gas concentration and isotope monitoring: optimization and integration of novel soil gas probes with online trace gas detection

Abstract. Gas concentrations and isotopic signatures can unveil microbial metabolisms and their responses to environmental changes in soil. Currently, few methods measure in situ soil trace gases such as the products of nitrogen and carbon cycling or volatile organic compounds (VOCs) that constrain microbial biochemical processes like nitrification, methanogenesis, respiration, and microbial communication. Versatile trace gas sampling systems that integrate soil probes with sensitive trace gas analyzers could fill this gap with in situ soil gas measurements that resolve spatial (centimeters) and temporal (minutes) patterns. We developed a system that integrates new porous and hydrophobic sintered polytetrafluoroethylene (sPTFE) diffusive soil gas probes that non-disruptively collect soil gas samples with a transfer system to direct gas from multiple probes to one or more central gas analyzer(s) such as laser and mass spectrometers. Here, we demonstrate the feasibility and versatility of this automated multiprobe system for soil gas measurements of isotopic ratios of nitrous oxide (δ18O, δ15N, and the 15N site preference of N2O), methane, carbon dioxide (δ13C), and VOCs. First, we used an inert silica matrix to challenge probe measurements under controlled gas conditions. By changing and controlling system flow parameters, including the probe flow rate, we optimized recovery of representative soil gas samples while reducing sampling artifacts on subsurface concentrations. Second, we used this system to provide a real-time window into the impact of environmental manipulation of irrigation and soil redox conditions on in situ N2O and VOC concentrations. Moreover, to reveal the dynamics in the stable isotope ratios of N2O (i.e., 14N14N16O, 14N15N16O, 15N14N16O, and 14N14N18O), we developed a new high-precision laser spectrometer with a reduced sample volume demand. Our integrated system – a tunable infrared laser direct absorption spectrometry (TILDAS) in parallel with Vocus proton transfer reaction mass spectrometry (PTR-MS), in line with sPTFE soil gas probes – successfully quantified isotopic signatures for N2O, CO2, and VOCs in real time as responses to changes in the dry–wetting cycle and redox conditions. Broadening the collection of trace gases that can be monitored in the subsurface is critical for monitoring biogeochemical cycles, ecosystem health, and management practices at scales relevant to the soil system.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS1988

Bioisolation on the Space Station

Animal research on the Space Station presents the need for bioisolation, which is here defined as instrumental and operational provisions, which will prevent the exchange of particles greater than 0.3-micron size and microorganisms between crew and animals. Current design principles for the Biological Research Project thus call for: (1) use of specific pathogen-free animals; (2) keeping animals at all times in enclosed habitats, provided with microbial filters and a waste collection system; (3) placing habitats in a holding rack, centrifuge, and workbench, all equipped with particulate and odor filters, (4) washing dirty cage units in an equipment cleaner, with treatment and recycling of the water; (5) designing components and facilities so as to ensure maximal accessibility for cleaning; and (6) defining suitable operational procedures. Limited ground tests of prototype components indicate that proper bioisolation can thus be achieved.

Bonting, Sjoerd L.↗

NASA NTRS2010

Mutualism in a Reduced Gravity Environment (MuRGE)

Mutualism in a Reduced Gravity Environment (MuRGE) is a ground research study to determine the feasibility of assessing fungi-plant (Piriformospora indica-Arabidopsis thaliana) interactions in microgravity. Seeds from the plant Arabiddospsis thaliana (At) will be grown in the presence of Piriformospora indica (Pi) an endophytic Sebacinacae family fungus. Pi is capable of colonizing the roots of a wide variety of plant species, including non-mycorrhizal hosts like At, and promoting plant growth similarly to AMF (arbusuclar mychorrizal fungi) unlike most AMF, Pi is not an obligate plant symbiont and can be grown in the absence of a host. In the presence of a suitable plant host, Pi can attach to and colonize root tips. Interaction visualization is accomplished with strong autofluorescence in the roots, followed by root colonization via fungal hyphae, and chlamydospore production. Increased root growth can be observed even before root colonization is detectable. In addition, Pi chlamydospores generated from axenic culture in microgravity will be used to inoculate roots of At grown in 1g to determine the effect of microgravity upon the inherent virulence or beneficial effects. Based on recent reports of increased virulence of S. typhimurium, P. aeruginosa, and S. Pneumoniae in reduced gravity, differences in microbial pathogenic responses and host plant systemic acquired resistance are expected. The focus of this project within MuRGE involved the development P. indica culture media evaluation and microscopy protocol development. High, clean spore harvest yields for the detection of fungi-plant interactions microscopically was the immediate goal of this experiment.

Haire, Timothy C.↗

NASA NTRS2012

From Kennedy, to Beyond: Growing Plants in Space

Astronauts cannot have their cake and eat it too, but what about growing a salad and eating it? As NASA continues to push the envelope on Space exploration and inhabitance the need for a fresh food source becomes more vital. The Life Support team at NASA is using a system developed by ORBITEC the VEGGIE, in which astronauts aboard the ISS, and potentially the Moon and Mars, will be capable of growing food. The introduction of plants not only gives astronauts a means of independently supplying food, but also recreation, oxygen replenishment and psychological benefits. The plants were grown in "pillows", the system used for growing plants within the VEGGIE. This test included 4 types of media mixtures that are composed of a clay based media called Arcilite and Fafard #2, which is a peat moss-based media ( <1 mm Arcilite, 1-2 mm of Arcilite, 1:1 <1 mm & 1-2 mm mixture and 1:1 Arcilite & Fafard mixture). Currently, 3 lettuce cultivars are being grown in 4 mixtures of media. Tests were being conducted to see which form of media has the ratio of best growth and least amount of microbes that are harmful. That is essential because a person's body becomes more susceptible to illness when they leave Earth. As a result, test must be conducted on the "pillow" system to assess the levels of microbial activity. The cultivars were tested at different stages during their growing process for microbes. Datum show that the mix of Fafard and Arcilite had the best growth, but also the most microbes. This was due to the fact that Fafard is an organic substance so it contains material necessary for microbes to live. Data suggest that the <1 mm Arcilite has an acceptable amount of growth and a lower level of microbes, because it is non-organic.

Flemming, Cedric, II↗

NASA NTRSDate not supplied

Early Results from a Broad Compatibility Study of Various Materials with Ionic Silver Biocide

Ionic silver is baselined for microbial control in spacecraft potable water systems forfuture exploration missions, but materials compatibility analysis is required to evaluate thepassive depletion of ionic silver concentration onto wetted material surfaces over time.Various articles concerning such testing have been published that examine interactions withwater containing ionic silver biocide, but most tests have focused on only a couple ofmaterials each and comparing results of different evaluations to one another has provedchallenging.This paper reports the first results from static exposure testing of a large arrayof material coupons to a 400 parts per billion (ppb) aqueous silver fluoride (AgF) solution,using a surface to volume ratio of approximately 2 cm-1. The test is designed in two mainstages. Stage 1 is a one-week screening to evaluate silver uptake. Materials that performmodestly to well after that week are promoted to Stage 2, which is a longer test with periodicsampling to examine the silver uptake rates over time; these samples are evaluated for otherwater quality parameters in addition to the remaining silver concentration. In a tangentialinvestigation, select materials that take up some silver in Stage 1 may be “aged” by repeatingthe Stage 1 test to determine whether repeated exposure reduces silver uptake rate, andsuccessfully aged materials may then continue to Stage 2 testing. The materials under testinclude metallic and polymeric materials with various surface finishes, treatments, andcoatings, as well as select other materials historically used in spacecraft water systems. Thistest began in August 2019, and thus only includes early results; future follow-on papers willinclude additional results as the test progresses. The ultimate goal builds a broad, easilycomparable data set that can be used to guide material selections for silver biocide-compatible spacecraft water system design.

Silver Biocide↗

NASA NTRSDate not supplied

Mitigation of Silver Ion Loss from Solution by Polymer Coating of Metal Surfaces, Part IV

Ionic silver (Ag+) biocide is a leading candidate to provide residual microbial control in spacecraft potable water systems, but suffers from rapid concentration loss due to interactions with the metallic storage containers and tubing. One approach to resolve this problem is the coating of metal surfaces with an inert barrier film. In previous reports, we have described our investigations addressing Ag+ loss mitigation and adhesion performance of parylene barrier coatings on coupons of several metal alloys and 316Ltubing under static immersion. In such conditions, parylene-C and -AF4 coatings have shown excellent Ag+loss mitigation and mixed long-term adhesion performance, depending on parylene species and substrate surface chemistry/structure. In Part IV of this series, we report on our work to investigate the performance of parylene-C barrier coatings, under more challenging and realistic conditions, in order to evaluate potential suitability for use. The resilience and associated Ag+ loss with Parylene-C coating on½” (12.7 mm)316L tubing and fittings under medium-term immersion and repeated fitting dis/reassembly were investigated. Potential mechanical challenges to barrier coatings in spacecraft potable water systems were investigated, with two main focuses being identified: liquid flow/pressure cycling in tubing and cyclic operation of bellows in positive-expulsion storage tanks. Two corresponding testbeds for experimental characterization of coating performance were developed. In the Flow/Pressure Testbed System, Ag+ solution will be pumped through internally coated 316L tubing, with optional pressure cycling. In the Bellows Tank Testbed System, an internally coated edge-welded 316L bellows containing Ag+ solution will be cyclically extended and compressed, analogous to the operation of bellows tanks used for potable water storage and delivery on the International Space Station. The design and operation of these testbeds are described. Finally, the Ag+ adsorption and saturation behaviors of Kalrez and Viton, for potential use as seal materials in fittings and other connections, were characterized in limited experiments.

mitigation↗

DOE OSTI2025

Development and flight-testing of modular autonomous cultivation systems for biological plastics upcycling aboard the ISS

Cultivation of microorganisms in space has enormous potential to enable in-situ resource utilization (ISRU) Here, we develop an autonomous payload with fully programmable serial passaging and sample preservation, termed the Modular Open Biological Platform (MOBP), and flight-test the MOBP aboard the International Space Station (ISS) by conducting enzymatic and microbial plastics upcycling experiments. The MOBP is a compact, modular bioreactor system that allows for sustained microbial growth via automated media transfers, such as those for sample collection and storage for terrestrial analyses, and precise data monitoring from integrated sensors. The MOBP was flight-tested with two experiments designed to evaluate biological upcycling of the plastic poly(ethylene terephthalate) (PET). The bioproduct βKA can be polymerized into a nylon-6,6 analog with improved properties for use in the production of a variety of materials. We posit the MOBP will aid in democratizing the execution of synthetic biology in spaceflight towards enabling ISRU.

09 BIOMASS FUELS↗

DOE OSTI2025

Interrelationships among methods of estimating microbial biomass across multiple soil orders and biomes

Understanding the role of soil microbes is critical to ecosystem processes, and more thorough comparisons of measurement proxies for soil microbial biomass could broaden the inclusion of explicit microbial parameterization in soil carbon cycling and earth system models. We measured physical, chemical, and biological data from eight soil orders representing 11 major biomes and four climate regions. Four prominent methods to measure microbial abundance—chloroform fumigation extraction (CFE), total DNA yield, gene copy number by quantitative polymerase chain reaction (GCN), and phospholipid fatty acids (PLFA)—were compared to assess their relationships with each other and with soil characteristics. Correlations were observed when comparing methods, with CFE correlating strongly with total DNA yield, GCN, and PLFA; CFE with bacterial GCN and bacterial PLFA; and to a lesser extent, total PLFA and total DNA yield. Correlations improved with the removal of organic soils (Histosols, Gelisols). Comparisons involving extracted DNA were improved by correcting for clay content, due to DNA extraction inefficiencies in clay-rich soils. Correlations involving fungi (PLFA or GCN) were always less significant. These methods could serve as reliable, inter-relatable proxies for the estimation of total soil microbial biomass while recognizing that the proxies are less effective at parsing differences between bacteria and fungi. Here, we provide specific equations to relate measures of soil microbial biomass by these four different methods to enable microbial models to utilize a greater diversity of observed data sources in parameterizations and simulations. Caveats for the equations and their values are also discussed.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Data from: Temporal dynamics of free‐living nitrogen fixation in the switchgrass rhizosphere

Here we present data associated with the manuscript, Temporal dyanmics of free-living nitrogen fixation in the switchgrass rhizosphere. Free-living nitrogen fixation (FLNF) represents an important terrestrial N source and is gaining interest for its potential to contribute plant available N to bioenergy cropping systems. Switchgrass, a cellulosic bioenergy crop, may be reliant on FLNF when particularly when grown on low N marginal lands. These potential contributions of FLNF to switchgrass and the controls on this process are not well understood. We evaluated drivers of FLNF rates and N-fixing microbial community composition in field-grown switchgrass systems over two years with high temporal sampling. We found climate variables to be strong drivers of FLNF in switchgrass systems, compared to other environmental and biological factors including soil nutrients and N-fixing microbial community composition. Increased soil moisture availability tended to promote FLNF, but extreme rainfall events appeared to be detrimental. These climate-related responses suggest FLNF-derived N contributions may be reduced under projected climate shifts. We also found a significant, but weak correlation between N-fixing microbial community composition and FLNF rates including an observed shift in community composition between 2017 and 2018 and a similarly significant difference in FLNF rates between years. Lastly, we found that seasonal FLNF N contributions, based on measurement with high temporal resolution, may meet up to 80% of switchgrass N demands.

09 BIOMASS FUELS↗

DOE OSTI2021

Legacy Effects of Intercropping and Nitrogen Fertilization on Soil N Cycling, Nitrous Oxide Emissions, and the Soil Microbial Community in Tropical Maize Production

Maize-forage grasses intercropping systems have been increasingly adopted by farmers because of their capacity to recycle nutrients, provide mulch, and add C to soil. However, grasses have been shown to increase nitrous oxide (N 2 O) emissions. Some tropical grasses cause biological nitrification inhibition (BNI) which could mitigate N 2 O emissions in the maize cycle but the reactions of the N cycle and the microbial changes that explain the N 2 O emissions are little known in such intercropping systems. With this in mind, we explored intercropping of forage grasses ( Brachiaria brizantha and Brachiaria humidicola ) with distinct BNI and yield potential to increase N cycling in no-till maize production systems compared to monocrop with two N rates (0 and 150 kg ha −1 ) applied during the maize season. These grasses did not strongly compete with maize during the period of maize cycle and did not have a negative effect on grain yield. We observed a legacy of these grasses on N mineralization and nitrification through the soil microbiome during maize growth. We observed that B. humidicola , genotype with higher BNI potential, increased net N mineralization by 0.4 mg N kg −1 day −1 and potential nitrification rates by 1.86 mg NO 3 -N kg −1 day −1 , while B. brizantha increased the soil moisture, fungi diversity, mycorrhizal fungi, and bacterial nitrifiers, and reduced saprotrophs prior to maize growth. Their legacy on soil moisture and cumulative organic inputs (i.e., grass biomass) was strongly associated with enhanced mineralization and nitrification rates at early maize season. These effects contributed to increase cumulative N 2 O emission by 12.8 and 4.8 mg N 2 O-N m −2 for maize growing after B. brizantha and B. humidicola , respectively, regardless of the N fertilization rate. Thus, the nitrification inhibition potential of tropical grasses can be outweighed by their impacts on soil moisture, N recycling, and the soil microbiome that together dictate soil N 2 O fluxes.

Canisares, Lucas P.↗

NASA NTRSDate not supplied

Tracking Metabolic Changes in Microbial Culture using Redox Measurements

During long-term space missions, microbial cultures accumulate the effects of low-dose radiation, microgravity, and other factors; altered growth and metabolic activity may occur before viability effects. This could affect functionality of bioreactors or other bio-enabled mission systems, as well as shed light on human health. Spaceflight microbiology studies beyond the low Earth orbit exposure afforded by the ISS have been limited. Nanosatellites offer an increasingly popular alternative for deep space missions. However, the communications delay requires biofluidic automation of a pre-defined experimental protocol, and the lack of sample return (reliance on sensors in flight) can significantly limit feasible investigations. Previous biological CubeSats (PharmaSat, O/OREOS, EcAMSat) have used alamarBlue, an off-the-shelf formulation of the redox indicator dye resazurin, to track metabolic activity, as will BioSentinel, the upcoming interplanetary microbiology experiment. A series of ground experiments (see abstracts by Liddell, Santa Maria, and A. Kim) were conducted using a microbial culture system outfitted with an electrochemical sensor array (electrical conductivity, pH, oxidation-reduction potential, and dissolved oxygen) with alamarBlue and the same strain of Saccharomyces cerevisiae as BioSentinel. By improving mapping of measured changes in alamarBlue kinetics to physicochemical changes, and ultimately to biological alterations such as shifted metabolic pathways, this work supplements data analyses from past missions and planning for future missions using alamarBlue to characterize space radiation effects. Initial results indicate that alamarBlue acts like a redox buffer; its presence significantly changes redox kinetics in otherwise identical cultures. The initial color change (blue resazurin reduced to red/pink resorufin) appears as a redox plateau. A second plateau, likely corresponding to the second color transition (resorufin to the colorless hydroresorufin), occurs at a lower redox value. The relationship to carbon source exhaustion, dissolved oxygen depletion, cell death, and measured redox potential is complex and still under study.

Tracking↗

DOE OSTI2022

Dynamic Responses of Trace Metal Bioaccessibility to Fluctuating Redox Conditions in Wetland Soils and Stream Sediments

Natural aquatic systems undergo fluctuating redox conditions due to microbial activity, varying water saturation levels, and nutrients dynamics. With fluctuating redox conditions, trace metals can mobilize or sequester in response to changes in iron and sulfur speciation and the concentrations and lability of organic carbon. This study examined the effect of redox fluctuations on trace metal mobility in samples collected from two different natural aquatic systems: riparian wetlands and a stream. The wetland soils contained low sulfur and total Fe contents as compared to stream sediments. The mineral composition at both sites was dominated by quartz. We incubated water-saturated soils under three cycles of anoxic-oxic conditions (τanoxic:τoxic = 3) spanning 24 days and monitored the change in dissolved and bioavailable metal concentrations. For both natural systems, reduction of iron oxides under anoxic conditions caused Co and Zn release. In contrast, oxidation of sulfides mobilized Cu under oxic conditions in both sites. In wetland soils, dissolution of Fe (hydr)oxides increased Ni solubility; however, in stream sediments, Ni release occurred when sulfides or organic matter were oxidized. For stream sediments, each subsequent redox cycle increased the bioavailability of trace metals. Redox fluctuations in wetland soils increased bioavailable Zn and Cu and decreased bioavailable Ni and Co. This study illustrates that different trace metals display distinct bioavailability patterns during redox fluctuations in natural environments. The biogeochemical cycling of nutrients in systems with redox fluctuations may be influenced by these trace metal availability patterns in addition to the availability of electron donors and acceptors.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

NASA NTRS2013

Microbial Monitoring of the International Space Station

Humans living and working in the harsh environment of space present many challenges for habitability engineers and microbiologists. Spacecraft must provide an internal environment in which physical (gas composition, pressure, temperature, and humidity), chemical, and biological environmental parameters are maintained at safe levels. Microorganisms are ubiquitous and will accompany all human-occupied spacecraft, but if biological contamination were to reach unacceptable levels, long-term human space flight would be impossible. Prevention of microbiological problems, therefore, must have a high priority. Historically, prevention of infectious disease in the crew has been the highest priority, but experience gained from the NASA-Mir program showed that microbial contamination of vehicle and life-support systems, such as biofouling of water and food, are of equal importance. The major sources of microbiological risk factors for astronauts include food, drinking water, air, surfaces, payloads, research animals, crew members, and personnel in close contact with the astronauts. In our efforts to eliminate or mitigate the negative effects of microorganisms in spacecraft, the National Aeronautics and Space Administration (NASA) implemented comprehensive microbial analyses of the major risk factors. This included the establishment of acceptability requirements for food, water, air, surfaces, and crew members. A robust monitoring program was then implemented to verify that the risks were within acceptable limits. Prevention of microbiological problems is preferred over mitigation of problems during flight, and preventive steps must begin very early in the design phase. Spacecraft development must include requirements to control free water from humidity, condensate, hygiene activities, and other releases. If water is available, microbes are likely to grow because sufficient nutrients are potentially available. Materials selected for the spacecraft must not promote or support microbial growth. Air filtration can dramatically reduce the number of airborne bacteria, fungi, and particulates in spacecraft breathing air. Waterborne bacteria can be reduced to acceptable levels by thermal inactivation of bacteria during water processing, along with a residual biocide, and filtration at the point of use can ensure safety. System design must include onboard capability to achieve recovery of the system from contamination. Robust housekeeping procedures that include periodic cleaning and disinfection will prevent high levels of microbial growth on surfaces. Food for consumption in space must be thoroughly tested for excessive microbial content and pathogens before launch. Thorough preflight examination of flight crews, consumables, payloads, and the environment can greatly reduce pathogens in spacecraft. Many of the lessons learned from the Space Shuttle and previous programs were applied in the early design phase of the International Space Station, resulting in the safest space habitat to date. This presentation describes the monitoring program for the International Space Station and will summarize results from preflight and on-orbit monitoring.

Pierson, Duane L.↗