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At least 343 records · Page 19
All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

Reproducible growth of Brachypodium in EcoFAB 2.0 reveals that nitrogen form and starvation modulate root exudation

Understanding plant-microbe interactions requires examination of root exudation under nutrient stress using standardized and reproducible experimental systems. We grew Brachypodium distachyon hydroponically in fabricated ecosystem devices (EcoFAB 2.0) under three inorganic nitrogen forms (nitrate, ammonium, and ammonium nitrate), followed by nitrogen starvation. Analyses of exudates with liquid chromatography–tandem mass spectrometry, biomass, medium pH, and nitrogen uptake showed EcoFAB 2.0’s low intratreatment data variability. Furthermore, the three inorganic nitrogen forms caused differential exudation, generalized by abundant amino acids–peptides and alkaloids. Comparatively, nitrogen deficiency decreased nitrogen-containing compounds but increased shikimates-phenylpropanoids. Subsequent bioassays with two shikimates-phenylpropanoids (shikimic and p-coumaric acids) on soil bacteria or Brachypodium seedlings revealed their distinct capacity to regulate both bacterial and plant growth. Our results suggest that (i) Brachypodium alters exudation in response to nitrogen status, which can affect rhizobacterial growth, and (ii) EcoFAB 2.0 is a valuable standardized plant research tool.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Biofilm mitigation in hybrid chemical-biological upcycling of waste polymers

Accumulation of plastic waste in the environment is a serious global issue. To deal with this, there is a need for improved and more efficient methods for plastic waste recycling. One approach is to depolymerize plastic using pyrolysis or chemical deconstruction followed by microbial-upcycling of the monomers into more valuable products. Microbial consortia may be able to increase stability in response to process perturbations and adapt to diverse carbon sources, but may be more likely to form biofilms that foul process equipment, increasing the challenge of harvesting the cell biomass. To better understand the relationship between bioprocess conditions, biofilm formation, and ecology within the bioreactor, in this study a previously-enriched microbial consortium (LS1_Calumet) was grown on (1) ammonium hydroxide-depolymerized polyethylene terephthalate (PET) monomers and (2) the pyrolysis products of polyethylene (PE) and polypropylene (PP). Bioreactor temperature, pH, agitation speed, and aeration were varied to determine the conditions that led to the highest production of planktonic biomass and minimal formation of biofilm. The community makeup and diversity in the planktonic and biofilm states were evaluated using 16S rRNA gene amplicon sequencing. Results showed that there was very little microbial growth on the liquid product from pyrolysis under all fermentation conditions. When grown on the chemically-deconstructed PET the highest cell density (0.69 g/L) with minimal biofilm formation was produced at 30°C, pH 7, 100 rpm agitation, and 10 sL/hr airflow. Results from 16S rRNAsequencing showed that the planktonic phase had higher observed diversity than the biofilm, and that Rhodococcus, Paracoccus, and Chelatococcus were the most abundant genera for all process conditions. Biofilm formation by Rhodococcus sp. And Paracoccus sp. Isolates was typically lower than the full microbial community and varied based on the carbon source. Ultimately, the results indicate that biofilm formation within the bioreactor can be significantly reduced by optimizing process conditions and using pure cultures or a less diverse community, while maintaining high biomass productivity. The results of this study provide insight into methods for upcycling plastic waste and how process conditions can be used to control the formation of biofilm in bioreactors.

36 MATERIALS SCIENCE↗
DOE OSTI2024

A contactless in situ EFISH method for measuring electrostatic potential profile of semiconductor/electrolyte junctions

In photoelectrochemical cells, promising devices for directly converting solar energy into storable chemical fuels, the spatial variation of the electrostatic potential across the semiconductor–electrolyte junction is the key parameter that determines the cell performance. In principle, electric field induced second harmonic generation (EFISH) provides a contactless in situ spectroscopic tool to measure the spatial variation of electrostatic potential. However, the total second harmonic generation (SHG) signal contains the contributions of the EFISH signals of semiconductor space charge layer and the electric double layer, in addition to the SHG signal of the electrode surface. The interference of these complex quantities hinders their analysis. In this work, to understand and deconvolute their contributions to the total SHG signals, bias-dependent SHG measurements are performed on the rutile TiO2(100)–electrolyte junction as a function of light polarization and crystal azimuthal angle (angle of the incident plane relative to the crystal [001] axis). A quadratic response between SHG intensity and the applied potential is observed in both the accumulation and depletion regions of TiO2. The relative phase difference and amplitude ratio are extracted at selected azimuthal angles and light polarizations. At 0° azimuthal angle and s-in–p-out polarization, the SHG intensity minimum has the best match with the TiO2 flatband potential due to the orthogonal relative phase difference between bias-dependent and bias-independent SHG terms. We further measure the pH-dependent flatband potential and probe the photovoltage under open circuit conditions using the EFISH technique, demonstrating the capability of this contactless method for measuring electrostatic potential at semiconductor–electrolyte junctions.

Chemistry↗
NASA NTRS2003

Autotrophic Ecosystems on the Early Earth

Ophiolite sequences, sections of lower oceanic crust and upper mantle that have been thrust onto continental craton, are located in northern and central California and provide easily accessible areas that serve as good analogs for similar, more extensive areas of the early Earth. We have begun investigating and characterizing these sites in order to understand better the processes that may be responsible for the water chemistry, mineralogy and biology of similar environments on the early Earth. The geophysical and geochemical processes in these terranes provide niches for unique communities of extremeophiles and likely provide a good analog to the location that first gave rise to life on Earth. The ophiolites found in northern and central California include the Trinity, Josephine, Coast Range and Point Sal, all of which are approximately 160 million years old. Fluids from serpentinizing springs are generally alkaline with high pH and H2 contents, indicating that the mafic rock compositions control the fluid composition through water-rock reactions during relatively low-grade hydrothermal processes. There are significant amounts of primary mineralogy remaining in the rocks, meaning that substantial alteration processes are still occurring in these terranes. The general reaction for serpentinization of olivine is given by one of the authors. olivine + H2O = serpentine + brucite + magnetite + H2. We have analyzed the mineralogical composition of several rock samples collected from the Coast Range Ophiolite near Clear Lake, CA by electron microprobe. The remnant primary mineralogy is fairly urnform in composition, with an olivine composition of Fo(sub 90), and with pyroxene compositions of En(sub 90) for orthopyroxene and En(sub 49)Wo(sub 48)Fs(sub 03) for the clinopyroxene. Other primary phases observed include chromites and other spinels. Examination of petrographic thin sections reveals that serpentinization reactions have occurred in these locations. The serpentine resulting from aqueous alteration of olivine resides in veins that are see to cross cut the primary mineral grains. There are several generations of alteration products, comprised mostly of serpentines that are magnesium rich, with magnetite, brucite and carbonates observed as accessory minerals. The formation of carbonates can be taken to indicate the presence of CO2 in the altering fluids. We collected samples from a spring in the Coast Range Ophiolite in order to determine whether the geochemical environment serves as a habitat for chemotrophic microorganisms. DNA was extracted from the sediment samples and the 16s rRNA gene was PCR amplified using universal Archaeal primers. Denaturing gradient gel electrophoresis (DGGE) was used to determine the community of Archaea thriving in these samples. Our results indicate that there were 8 different genera of Archaea from a single sample. A sequence was obtained from one of these eight. The sequence is of an organism similar to Halorubrum tibetense, and alkalophilic Archaeon. This result suggests that these environments are likely hosts for communities of organisms that are adapted for the unique chemistry provided by the alkaline spring.

Schulte, M.↗
DOE OSTI2020

Anaerobic respiration pathways and response to increased substrate availability of Arctic wetland soils

The availability of labile carbon (C) compounds in Arctic wetland soils is expected to increase due to thawing permafrost and increased fermentation as a result of decomposition of organic matter with warming. How microbial communities respond to this change will affect the balance of CO 2 and CH 4 emitted during anaerobic organic matter decomposition, and ultimately the net radiative forcing of greenhouse gas emissions from these soils. While soil water content limits aerobic respiration, the factors controlling methanogenesis and anaerobic respiration are poorly defined in suboxic Arctic soils. In this study, we conducted incubation experiments on two tundra soils from field sites on the Seward Peninsula, Alaska, with contrasting pH and geochemistry to determine the pathways of anaerobic microbial respiration and changes with increasing substrate availability upon warming. In incubation of soils from the circumneutral Teller site, the ratio of CO 2 to CH 4 dropped from 10 to <2 after 60 days, indicating rapid depletion of alternative terminal electron acceptors (TEAs). Addition of acetate stimulated production of CO 2 and CH 4 in a nearly 1:1 ratio, consistent with methanogenesis, and the composition of the microbial community shifted to favor clades capable of utilizing the added acetate such as the Fe(III)-reducing Geobacter and the methanogenic archaea Methanosarcina. In contrast, both CO 2 and CH 4 production declined with acetate addition during incubation of soils from the more acidic Council site, and fermentative microorganisms increased in abundance despite the high availability of fermentation products. These results demonstrate that the degree to which increasing substrate availability stimulates greenhouse gas production in tundra wetlands will vary widely depending on soil pH and geochemistry.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS2000

High relative humidity increases yield, harvest index, flowering, and gynophore growth of hydroponically grown peanut plants

Growth chamber experiments were conducted to study the physiological and growth response of peanut (Arachis hypogaea L.) to 50% and 85% relative humidity (RH). The objective was to determine the effects of RH on pod and seed yield, harvest index, and flowering of peanut grown by the nutrient film technique (NFT). 'Georgia Red' peanut plants (14 days old) were planted into growth channels (0.15 x 0.15 x 1.2 m). Plants were spaced 25 cm apart with 15 cm between channels. A modified half-Hoagland solution with an additional 2 mM Ca was used. Solution pH was maintained between 6.4 and 6.7, and electrical conductivity (EC) ranged between 1100 and 1200 microS cm-1. Temperature regimes of 28/22 degrees C were maintained during the light/dark periods (12 hours each) with photosynthetic photon flux (PPF) at canopy level of 500 micromoles-m-2s-1. Foliage and pod fresh and dry weights, total seed yield, harvest index (HI), and seed maturity were greater at high than at low RH. Plants grown at 85% RH had greater total and individual leaflet area and stomatal conductance, flowered 3 days earlier and had a greater number of flowers reaching anthesis. Gynophores grew more rapidly at 85% than at 50% RH.

NASA Discipline Life Support Systems↗
NASA NTRS1980

Studies of the chemical basis of the origin of protein synthesis Initiation and direction of peptide growth

The data presented in this paper show that the ease of nonenzymatic activation of carboxylic acids by ATP at pH 5 varies directly with the pKa of the carboxyl group, and is consistent with the idea that it is the protonated form of the carboxyl group which participates in the activation reaction. Consequently, since most N-blocked amino acids have higher pKas than do their unblocked forms, they are activated more readily, and it has been demonstrated that this principle applies to peptides as well, which are activated more rapidly than single amino acids. It is proposed that this fact may be partly responsible for the origin of two important features still observed in contemporary protein synthesis: (1) initiation in prokaryotes is accomplished with an N-blocked amino acid, and (2) elongation in all living systems occurs at the carboxyl end of the growing peptide.

Mullins, D. W., Jr.↗
DOE OSTI2023

N-induced soil acidification triggers metal stimulation of soil methane oxidation in a temperate steppe ecosystem

Methane (CH 4 ) is a potent greenhouse gas, and it is well established that low nitrogen (N) stimulates- and high N suppresses CH 4 oxidation in grassland ecosystems. Here, in this study, we examined the response of CH 4 uptake to long-term (>10 years) multi-level N additions in a temperate steppe of northern China. The N impacts on CH 4 uptake transitioned from positive to negative at the N addition rate of 4 g N m -2 y -1 . However, the high-N suppression on CH 4 uptake was partially relieved when the continuous (>10 years) high N inputs (16 g N m -2 y -1 to 64 g N m -2 y -1 ) caused soil pH drop to < 5.7. Further experiments revealed that continuous high-N inputs acidified soil to pH < 5.7 that released metal ions to stimulate monooxygenase enzyme activity; therefore, the CH 4 oxidation suppression was partially relieved under continuous N loading. Structural equation model results confirmed that metal ions, such as Iron (Fe 3+ ), Manganese (Mn 4+ ), and Copper (Cu) desorbed by soil particles, were negatively correlated with soil pH while positively correlated with CH 4 oxidation under high N inputs. These analyses suggested that continuous high N inputs-induced soil acidification can release previously bounded metal elements that partially alleviated the N suppression on CH 4 uptake. The N-induced soil acidification impacts on CH 4 uptake might play a critical role in global CH 4 cycling and deserve further investigation as external N inputs continue climbing.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRSDate not supplied

Advancing Satellite-Constrained Modeled Air-Sea CO 2 Fluxes With a Focus on the Strength of the Southern Ocean Carbon Sink

Challenge and Motivation: The ocean plays a critical role in mitigating climate change by removing approximately a quarter of annual anthropogenic CO 2 emissions from the atmosphere. Model-based estimates point to the Southern Ocean as a key marine region, responsible for approximately 40 % of the anthropogenic carbon uptake by the global ocean. However, the contemporary strength of the Southern Ocean carbon sink has recently come into question. On the one hand, airborne-based observations of atmospheric CO 2 gradients indicate that the Southern Ocean represents a strong net sink of atmospheric CO 2 , consistent in magnitude with atmospheric inversion estimates and surface-ocean partial pressure of CO 2 (pCO 2 )-based products. On the other hand, estimates of pCO 2 based on in situ pH measurements taken by biogeochemical profiling floats yield strong wintertime outgassing fluxes that greatly reduce the Southern Ocean’s annually integrated CO 2 uptake. This uncertainty in the strength of the Southern Ocean air-sea CO 2 flux and its role in the global carbon cycle hinders our ability to constrain global carbon fluxes, one of the major goals of NASA’s Carbon Monitoring System (CMS). Opportunity: The NASA Ocean Biogeochemical Model (NOBM) produces near-global pCO 2 and air-sea CO 2 flux estimates that are currently included into the NASA’s Goddard Earth Observing System (GEOS) models in support of the CMS effort to monitor global carbon fluxes. The NOBM assimilates ocean color data to improve the representation of biogeochemical fluxes and overcome spatial and temporal gaps in the space-based retrievals. Here, we propose to advance the satellite-constrained flux estimates by investigating the uncertainties in the Southern Ocean air-sea CO 2 flux produced by the NOBM, and assess the value that remote sensing ocean color data can have in providing improved estimates of carbon fluxes in the ocean. Our proposed work includes the delivery of refined in situ float-based carbon fluxes to serve as a constraint on the model-based estimates. Taking advantage of the model’s integration of satellite ocean color data to represent multiple phytoplankton groups, we propose to deliver maps of biogenic carbon export specific to each modeled phytoplankton type and investigate the role of ecological plankton complexity in regulating marine carbon uptake and export. Goals: (a) Delivery of seasonally-adjusted float-based Southern Ocean air-sea CO 2 fluxes: We will produce updated and improved float-based air-sea CO 2 fluxes that will serve as a bias-reduced float-based constraint to evaluate our model-based estimates of the NOBM. (b) Investigation of uncertainties in Southern Ocean air-sea CO 2 flux from the NOBM: Modeled air-sea carbon fluxes will be evaluated against the updated float product as well as ship- and airborne-based data to identify uncertainties and potential model deficiencies. (c) Delivery of model-based carbon export partitioning by phytoplankton functional types (PFTs): We will produce depth-resolved maps of particulate organic export production integrated for all phytoplankton groups and allocated to each individual PFT in the model. The expected significance of this goal is to quantify the role that the functional-oriented diversity in phytoplankton groups represented in the NOBM plays in regulating air-sea CO 2 fluxes in the Southern Ocean.

Lionel A. Arteaga↗
DOE OSTI2020

Multidecadal shifts in forest plant diversity and community composition across glacial landforms in northern lower Michigan, USA

Understanding how plant community assemblage is affected by spatial and temporal patterns is crucial to understanding forest ecosystem responses to disturbance, including future climate change. In this article, we tracked how diversity and composition are distributed through space and time in a midsuccessional mixed hardwood forest in northern lower Michigan, United States. This region’s geographically and abiotically distinct glacial landforms influence both the spatial and temporal dynamics of its forest communities. Vegetation sampling plots (n = 87) were established at the University of Michigan Biological Station in 1990 and resampled in 2015. Vegetation in the overstory, sapling, and groundcover layers was censused. Abiotic variables, including elevation, pH, and soil nutrients, were measured in a subset of plots (n = 40). There were strong differences in diversity and community composition among glacial landforms, with the moraine having a 31% greater species richness in the groundcover layer compared with those of the other glacial landforms. Surprisingly, plant communities across all three vegetation layers showed little change over the 25-year period, and we found no evidence of differences in successional rates among glacial landforms. Our findings indicate that glacial landforms have a large influence on the production and maintenance of local plant diversity and community composition in this area and suggest that successional dynamics may manifest themselves over much longer time periods in these northern biomes.

Forestry↗
DOE OSTI2026

Disentangling High Harmonic Generation from Surface and Bulk States of a Topological Insulator

The discovery of topological phases has introduced a new dimension to materials science. Three-dimensional (3D) topological insulators (TIs) are a remarkable class of matter that is insulating in the bulk while hosting conductive topological surface states (TSSs) with unique charge and spin properties. High-order harmonic generation (HHG) has emerged as a powerful tool to probe condensed matter systems by providing insights into their electronic structure and dynamic behavior. Here, we investigate HHG in the prototype 3D-TI Bi$_2$Se$_3$. We demonstrate that the contributions of bulk and surface states to the harmonic emission can be controlled by tuning the thickness of thin film samples. An ultrathin (6 nm) film substantially enhances HHG from the surface states, while the bulk states dominate HHG in a thicker (50 nm) film. By applying a quasi-static terahertz perturbing field, we disentangle the bulk and surface responses and reveal the significant impact of the surface states' shift vector and Berry curvature on HHG. Our study provides effective methods for isolating the optical responses of TSSs from those of the bulk, which opens the door to resolving an ongoing debate regarding whether it is possible to reliably extract topological signatures in HHG.

Atomic Physics (physics.atom-ph)↗
NASA NTRS1991

The variability of the solar diameter

It is argued here that most of the variation in observations of the solar radius of over three centuries measure properties of the sun's limb darkening function and are affected by many sources of degradation of the solar image. They do not directly measure a true solar radius. From a long series of visual observations made by a single observer as well as recent photoelectric observations, evidence is found for periodicities in the apparent radius that occur in both modern and historical records. The magnitude of the observed variations is quite different in visual and photoelectric observations, suggesting that the process responsible for the periodicities is either one that modifies the solar limb darkening function or one that causes systematic variations in image blurring by the earth's atmosphere. A connection between solar magnetic activity an apparent radius seems likely, with evidence for such a relation dating back as fast as the Maunder minimum.

Ribes, E.↗
NASA NTRS2004

Optoelectronic Instrument Monitors pH in a Culture Medium

An optoelectronic instrument monitors the pH of an aqueous cell-culture medium in a perfused rotating-wall-vessel bioreactor. The instrument is designed to satisfy the following requirements: It should be able to measure the pH of the medium continuously with an accuracy of 0.1 in the range from 6.5 to 7.5. It should be noninvasive. Any material in contact with the culture medium should be sterilizable as well as nontoxic to the cells to be grown in the medium. The biofilm that inevitably grows on any surface in contact with the medium should not affect the accuracy of the pH measurement. It should be possible to obtain accurate measurements after only one calibration performed prior to a bioreactor cell run. The instrument should be small and lightweight. The instrument includes a quartz cuvette through which the culture medium flows as it is circulated through the bioreactor. The cuvette is sandwiched between light source on one side and a photodetector on the other side. The light source comprises a red and a green light-emitting diode (LED) that are repeatedly flashed in alternation with a cycle time of 5 s. The responses of the photodiode to the green and red LEDs are processed electronically to obtain a quantity proportional to the ratio between the amounts of green and red light transmitted through the medium.

Anderson, Melody M.↗
NASA NTRS2000

Effects of pH and dose on nasal absorption of scopolamine hydrobromide in human subjects

PURPOSE: The present study was conducted to evaluate the effects of formulation pH and dose on nasal absorption of scopolamine hydrobromide, the single most effective drug available for the prevention of nausea and vomiting induced by motion sickness. METHODS: Human subjects received scopolamine nasally at a dose of 0.2 mg/0.05 mL or 0.4 mg/0.10 mL, blood samples were collected at different time points, and plasma scopolamine concentrations were determined by LC-MS/MS. RESULTS: Following administration of a 0.2 mg dose, the average Cmax values were found to be 262+/-118, 419+/-161, and 488+/-331 pg/ mL for pH 4.0, 7.0, and 9.0 formulations, respectively. At the 0.4 mg dose the average Cmax values were found to be 503+/-199, 933+/-449, and 1,308+/-473 pg/mL for pH 4.0, 7.0, and 9.0 formulations, respectively. At a 0.2 mg dose, the AUC values were found to be 23,208+/-6,824, 29,145+/-9,225, and 25,721+/-5,294 pg x min/mL for formulation pH 4.0, 7.0, and 9.0, respectively. At a 0.4 mg dose, the average AUC value was found to be high for pH 9.0 formulation (70,740+/-29,381 pg x min/mL) as compared to those of pH 4.0 (59,573+/-13,700 pg x min/mL) and pH 7.0 (55,298+/-17,305 pg x min/mL) formulations. Both the Cmax and AUC values were almost doubled with doubling the dose. On the other hand, the average Tmax, values decreased linearly with a decrease in formulation pH at both doses. For example, at a 0.4 mg dose, the average Tmax values were 26.7+/-5.8, 15.0+/-10.0, and 8.8+/-2.5 minutes at formulation pH 4.0, 7.0, and 9.0, respectively. CONCLUSIONS: Nasal absorption of scopolamine hydrobromide in human subjects increased substantially with increases in formulation pH and dose.

Controlled Clinical Trial↗
DOE OSTI2020

Novel Foaming Solutions for High Level Waste (HLW) Processing - 20195

Foaming occurs during treatment of high level waste (HLW) at Savannah River Site's Defense Waste Processing Facility (DWPF) in the Chemical Process Cell (CPC) due to high gas generation from process steam and chemical offgas products [1]. The presence of amphiphilic particles in the waste slurry stabilizes the foam [2], [3]. Efficient processing of HLW requires foam control, as foam-overs lead to lower productivity and potential radioactive contamination of condensate streams. DWPF currently employs Siovation Antifoam 747, a surfactant produced by Momentive Performance Materials, as an antifoaming agent during waste treatment. Despite its ability to control foam generation, processing issues have arisen from its use [4]. During DWPF chemical processing, a chemical antifoam agent must be effective at temperatures up to 103 deg. C between pH 3-13. Antifoam 747 is effective at pH 7 but degrades as pH deviates [5]. Antifoam 747 is 90% by weight Momentive{sup TM} L77 and 10% by weight Momentive{sup TM} Y-17580. These ingredients are trisiloxane products, which are especially vulnerable to attack in acidic and caustic environments. Since Antifoam 747 readily degrades, periodic addition is required. Several processing concerns related to Antifoam 747 have affected DWPF operation. Its overuse warranted concern of potential flammable melter offgas under upset conditions due to the generation of carbon dioxide and hydrogen. As a result, a Potential Inadequacy in the Safety Analysis (PISA) was declared [6]. Antifoam contribution has been included in the melter offgas flammability strategy and the quantity of antifoam is monitored to ensure limits are not exceeded. Testing at Savannah River National Laboratory (SRNL) determined that propanal, tri-methylsilanol, and hexamethyldisiloxane are generated as Antifoam 747 degrades, leading to a second PISA [7]. The formation of these byproducts poses additional flammability risks. Furthermore, the methyl and ethyl groups of these compounds may be responsible for the formation of the methyl and ethyl mercury discovered in DWPF [8], [9]. The formation of organomercury has specifically been called out by DOE as a need for increased research. The effectiveness of non-chemical methods for foam control and the efficiency of alternative antifoam agents were evaluated as part of an effort to improve HLW treatment operations at DWPF and to reduce, or eliminate, the flammability hazards associated with Antifoam 747 currently in use. Foaming of high-level waste (HLW) slurries is an issue at the Defense Waste Processing Facility (DWPF) Chemical Process Cell (CPC) which is currently mitigated with a chemical antifoam agent. The effectiveness of non-chemical methods for foam control and the efficiency of alternative antifoams were examined to improve HLW treatment at DWPF and to eliminate the flammability hazards associated with Siovation Antifoam 747 currently in use. Non-chemical foam control methods were deemed unviable for chemical processing at DWPF, as the spray/mist technique requires too much water and space limitations make installation and implementation of headspace agitators unfeasible. The use of ultrasonic energy thickened and further stabilized the foam. Two suitable alternative defoaming agents, Momentive{sup TM} Y- 17112 and Evonik Surfynol{sup R} MD-20, were identified. Both defoamers control foam over a pH range of 4- 13 at boiling, with no flammable degradation products, and only minimal quantities (100-200 mg/kg) are required under glycolic acid conditions. In order to validate the effectiveness of these defoaming agents, SRNL recommends irradiation testing and laboratory scale SRAT/SME experiments with actual radioactive waste. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2023

High-Throughput Directed Evolution of Marine Microalgae and Phototrophic Consortia for Improved Biomass Yields (Final Report)

Primary project achievements include using selective pressures (O 2 , light, temperature) and developing culturing regimes for the diatom Nitzschia inconspicua str. hildebrandi to attain enrichments with an ~90% increase in areal biomass productivity relative to the parental strain under pond-mimicking conditions with high O 2 stress in laboratory bioreactors. The resulting strain (GAI-337) was tested further for dilution time, culture density, CO 2 supplementation, pH, temperature, and dissolved O 2 concentration under outdoor pond-mimicking conditions to improve areal productivities. These experiments yielded an optimum harvest and dilution time just after sunset, ~0.45 g AFDW L -1 initial culture density for maximal productivities, no requirement for CO 2 supplementation or pH control, maximal performance under a diel temperature curve going from 24 °C at night to 36 °C during the day, and benefits from some O 2 removal from the culture by bubbling with air. Using pond-mimicking laboratory bioreactors, N. inconspicua GAI-337 achieved ~42 g AFDW m -2 d -1 . Nutrient limitation experiments resulted in a biomass composition that equated to ~160 Gallons of Gasoline Equivalent energy per ton AFDW, highlighting the potential of GAI-337 as a promising renewable fuel feedstock strain. Genome resequencing has revealed genome alterations potentially contributing to the improved growth of GAI-337 in the laboratory. Based on the comparative analyses of the GAI-337 and GAI-229 (reference) strains, we identified 144 single nucleotide substitutions that resulted in amino acid change, 7 single nucleotide substitutions that resulted in protein truncation; 5 deletions; and 1 frameshift mutation. From the mutations that potentially affect expression of functionally annotated genes, particular interest was noted for an interferon-induced 6-16 family protein that may be involved in the host immune response against microbe invasion; the chaperone protein DnaK, which may function to protect the folding of proteins within the cell; and SPRY domain protein that is found in many eukaryotic proteins important in cell signaling pathways. Transcriptome analysis revealed over 1000 genes with increased transcript levels. Many of these and many of the genes with mutations are not yet functionally annotated and an increased bioinformatics effort is necessary to more completely analyze the Nitzschia inconspicua genome. Adaptive laboratory evolution (ALE) was performed for over 300 days using consecutive 0.5°C temperature increases in a constant temperature incubator to attain greater thermal tolerance in Nitzschia inconspicua. The adapted strain was able to grow at a constant temperature of 37.5°C; whereas this constant temperature was lethal to the parental control, which had an upper temperature boundary of 35.5°C prior to adaptive evolution. Several high-temperature clonal isolates were obtained from the evolved population following ALE, and increased temperature tolerance was observed in clonal adapted cultures. The final temperature adaptation was maintained through cryopreservation and was observed in multiple clonal isolates, including multiple clonal isolates with significantly increased cell size, indicating the potential occurrence of a sexual cycle during the clonal isolation process. A survey of Nannochloropsis strains was conducted for tolerances to high pH and high bicarbonate media. Nannochloropsis granulata showed promising growth in diel bioreactors and was successfully grown at the GAI Kauai farm site in long-term growth campaigns. Co-culturing using Nitzschia inconspicua, Nannochloropsis and a cyanobacterium were assembled in the laboratory to determine if productivity synergies could be attained. Although all strains grew well in the laboratory high-bicarbonate media individually, the cyanobacterium quickly outgrew the other strains in the laboratory consortium pushing the co-culture away from a diverse (and potentially synergistic assemblage) phototroph culture towards a monoculture dominated by the cyanobacterium. Several outdoor growth campaigns were conducted, with productivities ranging between 10-20 g/m 2 /d of biomass. The best performing strain in the laboratory (GAI-337) did not outperform reference strains at the Kauai farm under the conditions used. Addition growth campaigns are necessary under conditions that result in higher biomass (>20 g/m 2 /d) and that attain higher O 2 levels are likely necessary. Initial data indicate that the thermally adapted strain did slightly better than the control strain at higher temperatures; however, additional campaigns are necessary to establish statistical significance. In summary, Nitzschia inconspicua is able to attain exemplary biomass and lipid yields in the laboratory bioreactors. Strain evolution to both O 2 and temperature resulted in targeted strain improvements. Additional outdoor campaigns are necessary to determine if laboratory improvements translate to the field.

09 BIOMASS FUELS↗
DOE OSTI2024

Developing a Redox Network for Coastal Saltmarsh Systems in the PFLOTRAN Reaction Model

Coastal ecosystems have been largely ignored in Earth system models but are important zones for carbon and nutrient processing. Interactions between water, microbes, soil, sediments, and vegetation are important for mechanistic representation of coastal processes and ecosystem function. To investigate the role of these feedbacks, we used a reactive transport model (PFLOTRAN) that has the capability to be connected to the Energy Exascale Earth System Model (E3SM). PFLOTRAN was used to incorporate redox reactions and track chemical species important for coastal ecosystems as well as define simple representations of vegetation dynamics. Here our goal was to incorporate oxygen flux, salinity, pH, sulfur cycling, and methane production along with plant-mediated transport of gases and tidal flux. Using porewater profile and incubation data for model calibration and evaluation, we were able to create depth-resolved biogeochemical soil profiles for saltmarsh habitat and use this updated representation to simulate direct and indirect effects of elevated CO 2 and temperature on subsurface biogeochemical cycling. We found that simply changing the partial pressure of CO 2 or increasing temperature in the model did not fully reproduce observed changes in the porewater profile, but the inclusion of plant or microbial responses to CO 2 and temperature manipulations was more accurate in representing porewater concentrations. This indicates the importance of characterizing tightly coupled vegetation-subsurface processes for developing predictive understanding and the need for measurement of plant-soil interactions on the same time scale to understand how hotspots or moments are generated.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS1994

Distribution of cultivated and uncultivated cyanobacteria and Chloroflexus-like bacteria in hot spring microbial mats

Oligodeoxynucleotide hybridization probes were developed to complement specific regions of the small subunit (SSU) rRNA sequences of cultivated and uncultivated cyanobacteria and Chloroflexus-like bacteria, which inhabit hot spring microbial mats. The probes were used to investigate the natural distribution of SSU rRNAs from these species in mats of Yellowstone hot springs of different temperatures and pHs as well as changes in SSU rRNA distribution resulting from 1-week in situ shifts in temperature, pH, and light intensity. Synechococcus lividus Y-7c-s SSU rRNA was detected only in the mat of a slightly acid spring, from which it may have been initially isolated, or when samples from a more alkaline spring were incubated in the more acid spring. Chloroflexus aurantiacus Y-400-fl SSU rRNA was detected only in a high-temperature mat sample from the alkaline Octopus Spring or when lower-temperature samples from this mat were incubated at the high-temperature site. SSU rRNAs of uncultivated species were more widely distributed. Temperature distributions and responses to in situ temperature shifts suggested that some of the uncultivated cyanobacteria might be adapted to high-, moderate-, and low-temperature ranges whereas an uncultivated Chloroflexus-like bacterium appears to have broad temperature tolerance. SSU rRNAs of all uncultivated species inhabiting a 48 to 51 degrees C Octopus Spring mat site were most abundant in the upper 1 mm and were not detected below a 2.5-to 3.5-mm depth, a finding consistent with their possible phototrophic nature. However, the effects of light intensity reduction on these SSU rRNAs were variable, indicating the difficulty of demonstrating a phototrophic phenotype in light reduction experiments.

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