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At least 1,063 records · Page 59
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

SCALE 6.3 Validation: Spent Nuclear Fuel

This report is the fifth volume in a series documenting the validation of SCALE 6.3 with ENDF/B-VII.1 libraries for nuclear criticality safety, reactor physics, radiation shielding, and spent nuclear fuel applications. This fifth volume, which focuses on validating SCALE capabilities that impact spent nuclear fuel applications, provides an update of the similar validation reported for SCALE 6.2.4. The experimental data used as basis for validation herein consist of measurement data for nuclide inventories and decay heat, including the following: 1. radiochemical assay (RCA) measurements of nuclides important to burnup credit, decay heat, and radiation shielding in 205 light-water reactor (LWR) spent nuclear fuel samples that cover burnups ranging up to 80 GWd/MTU and initial fuel enrichments up to 4.9% 235U; 2. full-assembly decay heat measurements for 236 LWR assemblies with initial fuel enrichments ranging up to 4% 235 U, assembly burnups of 5–51 GWd/MTU, and decay times after fuel discharge in the 2- to 27-year range (of importance to spent nuclear fuel storage, transportation, and disposal); and 3. pulse fission irradiations for fissionable materials at cooling times of interest to severe accident analyses (<10 5 s).

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
DOE OSTI2025

Final Design for Thermal/Epithermal eXperiments (TEX) with Lithium Absorbers to Provide Validation Benchmarks for Y-12 Electrorefining Facility

One of the main goals of the Thermal/Epithermal eXperiments (TEX) project is to use existing Nuclear Criticality Safety Program (NCSP) assets to create critical experiment plutonium and uranium test beds for materials important to criticality safety that have insufficient benchmark evaluations. The plutonium test bed experiments were completed in 2018 and are published in the 2020 edition of the International Criticality Safety Benchmark Evaluation Project (ICSBEP) Handbook. The uranium test bed assemblies were completed in 2023 and accepted in the 2024 edition of the ICSBEP Handbook. The Nuclear Criticality Safety (NCS) group at Y-12 National Security Complex has identified programmatic need for validation cases for uranium electrorefining operations at Y-12. The electrorefining operation credits lithium enriched in 6 Li in addition to 35 Cl as absorbers in the design criticality safety evaluation for precluding criticality under upset conditions in the large and geometrically unfavorable electro-refiner. There is, however, inadequate experimental validation for the 6 Li absorbers. As an extension of the TEX uranium test bed, TEX-Cl critical experiments were performed with sodium chloride salt to address the 35 Cl thermal absorption as well as other validation needs at Los Alamos National Laboratory (LANL). These experiments were completed in 2024 and accepted into the 2025 ICSBEP Handbook. To continue the methodology used in TEX-Cl, TEX-Li aims to accomplish the same. The overall design of both experiments was to use commercially available, high purity, salts with polyethylene moderator and HEU plates to configure a critical assembly. Three experiments are planned for TEX-Li using encapsulated lithium carbonate (Li 2 CO 3 ), with natural 6 Li abundance. For all experiments, the highly enriched uranium (HEU) Jemima plates will be used as fissile material. Multiple layers will be stacked together with encapsulated Li 2 CO 3 alternated with polyethylene in standard configurations. Standard stacking was found to be optimal in matching the different sensitivity profiles provided by the Y-12 models. Three configurations are proposed with varying polyethylene moderation and a constant 1/4” absorber thickness. The first uses 11 layers of 5/4” polyethylene, the second uses 9 layers of 3/4” polyethylene, and the third uses 10 layers of 1/2” polyethylene. Calculations showed that some alternative forms of lithium-based materials provided slightly less-optimal sensitivity profiles when compared to lithium carbonate but come with other drawbacks. These alternatives included lithium aluminate (LiAlO 2 ), Aluminum-2050 alloy, Aluminum-8090, Aluminum-2095, lithium hydride (LiH), and lithium fluoride (LiF). Lithium aluminate and aluminum-2050 provided comparable sensitivity profiles when compared to lithium carbonate and can be used instead if lithium carbonate cannot be readily procured. After a broad material study, lithium carbonate outperformed any alternative material with a balance in affordability and workability. The assessment of experimental uncertainties of the non-absorber and absorber components was predicted to be 0.00089 and 0.00093 Δk eff , respectively. The largest uncertainties may be reduced with precision dimensional inspection of the components. Many of the parts and equipment for IER 575 have already been fabricated or procured for previous projects and therefore do not contribute significantly to the overall cost of this experiment. This includes the Jemima plates and Comet critical assembly machine, which are existing NCSP assets, as well as the aluminum platen and polyethylene reflector rings, which were fabricated and authorized for the TEX experiment involving HEU with polyethylene. Lithium carbonate containers will be procured by LANL and will be filled by LLNL. The total material costs for TEX-Li experiments are estimated to be on the order of $\$$47,400. Precision inspection, including dimensional, mass, density, and impurity, is recommended for all components for an estimated cost of $\$$12,000.

35Cl↗
DOE OSTI2024

HEU Metal Delayed Critical Experiments with 10 to 19 Inch Thick Graphite Reflectors

Approximately 100 graphite-reflected highly enriched uranium (HEU, 93.14 wt % 235 U) metal annular and cylindrical critical experiments were performed in the early 1960s at the Oak Ridge Critical Experiments Facility (ORCEF). This report presents details from experiment logbooks, experimental data sheets and the author's memory for 44 HEU metal (93.14 wt % 235 U) critical assemblies with graphite reflectors varying from 10 to 19 in. thick, outside diameters varying from 7 to 15 in., inside diameters varying from 7 to 13 in. and critical HEU metal masses varying from 20.4 to 69.0 kg. The data from the 44 experiments described in this report are acceptable for use as criticality safety benchmark experiments for the International Criticality Safety Evaluation Program (ICSBEP) once the uncertainty analysis on the measured k eff is completed. Based on previous ICSBEP benchmarks with this HEU metal at ORCEF, the uncertainties in the measured k eff are expected to be as low as ±0.0004. Preparation of this report is part of an effort at Oak Ridge National Laboratory (ORNL) to document more than 15 undocumented series of critical and subcritical experiments enumerated in Critical and Subcritical NEA Benchmark Possibilities for Measurements at ORCEF and Other US DOE Facilities (Mihalzo, ORNL/TM-2019/1188, 2019) and performed by ORNL at ORCEF and other US Department of Energy critical experiments facilities. More than 500 operational days of critical facility time were used, not including setup and dismantlement time. This documentation for a part of one series of graphite reflected highly enriched uranium metal critical experiments, that used 50 operational days of ORCEF time, was performed using funding received from the DOE Office of Nuclear Energy’s Nuclear Energy University Programs at the University of Tennessee Nuclear Engineering Department. This documentation was also supported by the Nuclear Criticality, Radiation Transport, and Safety programs at ORNL.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗
DOE OSTI2026

Liquid centrifugation-based isotope separation of 7 Li, 37 Cl and D

Liquid centrifugation is an emerging method to separate isotopes. By spinning a liquid containing target isotopes (e.g., pure chemical or solution), the heavier isotope will be enriched at the outer part of a centrifuge, and the lighter ones will be enriched at the inner part of a centrifuge. The separation capability is positively correlated with rotation speed, outer radius and mass difference between isotopes to separate. This award targets to understand whether liquid centrifugation is effective in separating isotopes important for nuclear fission technologies. The results include the following three sections: 1) Effects of solute concentration on isotope separation; 2) Preliminary progress of building a low-speed countercurrent centrifuge, and 3) Analysis on possible candidates for 7 Li and 37 Cl. The award trained one research scientist, one Ph.D. student, one master student, and one undergraduate on liquid centrifugation and isotope measurements.

07 ISOTOPE AND RADIATION SOURCES↗
DOE OSTI2026

Novel Method for Domestic Stable Isotope Production

This report presents the progress achieved on our chlorine isotope separation efforts in the CRADA #667 agreement. We used a microchannel distillation (MCD) packing structure to enrich HCl isotopes (H35Cl, H37Cl) and an isotachophoresis (ITP) separation to enrich ionic chloride species ( 35 Cl- and 37 Cl-). A 2 m tall MCD column was constructed and achieved 264 separation stages over several days, which is well above the goal of 50 stages outlined in the CRADA. The resulting height equivalent to a theoretical plate (HETP) was 0.76 cm. ITP tests were run ~ 24 hours and achieved a single-stage separation factor of 1.559.

07 ISOTOPE AND RADIATION SOURCES↗
DOE OSTI2026

Feasibility and strategic implications of deploying nuclear power reactors in Africa

This report assesses the feasibility and strategic implications of deploying nuclear power reactors, including large-scale plants, advanced small modular reactors (SMRs), and microreactors, in African countries. Case studies focus on South Africa, Egypt, Kenya, Ghana, and Nigeria, examining nuclear energy’s role in Africa’s rapidly evolving energy landscape, marked by fast-growing demand, significant electricity access gaps, increasing renewable penetration, and strong policy commitments to industrialization and energy security. Several U.S. reactor technologies and designs are considered based on their development status and readiness for deployment. The analysis finds that nuclear power can provide reliable, clean baseload and flexible generation, as well as high-temperature process heat for desalination, hydrogen production, and industrial applications. However, suitability is highly country-specific, depending on grid size and stability, transmission capacity, cooling water availability, regulatory readiness, and fuel supply chains. Near-term deployment opportunities are strongest for light-water reactors (such as NuScale, BWRX-300, AP300, and SMR-300) that use low-enriched uranium and build on proven technology. More advanced concepts, including gas-cooled, sodium-cooled, molten-salt cooled reactors, and microreactors, will likely be relevant for African deployment in the 2030s or later, contingent on demonstration projects, high-assay low-enriched uranium (HALEU) fuel availability, and mature international licensing frameworks. Economic analysis shows that SMRs are capital-intensive, with projected overnight costs for 300 MWe units in 2025 ranging from approximately 1.4 to 2.6 billion USD per module. The levelized cost of electricity (LCOE) is highly sensitive to the weighted average cost of capital (WACC). Given typically higher financing costs and utility balance-sheet weaknesses in many African countries, bankable project structures will require sovereign guarantees, robust offtake arrangements, and layered financing from export credit agencies, development finance institutions, and vendor nations. Comparisons with recent large nuclear projects in the United Arab Emirates (UAE) and Egypt underscore the central role of state-backed loans, long tenors, and concessional terms. Country case studies illustrate a spectrum of readiness and opportunity. South Africa operates two 920 MWe pressurized light water reactors (totaling 1,840 MWe) at Koeberg and has the most mature regulatory and industrial base, positioning it as a prime candidate for both large reactors and SMRs to replace coal, support desalination, and anchor industrial hubs. Egypt is constructing four VVER-1200 units at El Dabaa with strong state leadership and could later complement this fleet with SMRs for coastal and industrial applications. Kenya and Ghana are advancing through IAEA Milestones with growing institutional capacity and clear interest in SMRs that match their smaller grids and industrialization plans. Nigeria has the largest demand potential but faces acute constraints in grid reliability, project bankability, and regulatory capacity; targeted deployments of large reactors and SMRs near coastal or industrial sites could have high impact if accompanied by major grid upgrades and institutional reforms. The report identifies cross-cutting challenges such as financing, political continuity, public acceptance, nonproliferation and security, waste and back-end management, regulatory capacity, grid adequacy, and long deployment timelines for first-of-a-kind designs, and ANL/NSE-26/3 ii proposes broad directions for resolution. These include stronger multifaceted financing for nuclear, long-term national energy strategies that transcend electoral cycles, proactive stakeholder engagement, strengthened regional and national regulators, and systematic workforce development through centers of excellence and expanded training. The United States should develop partnerships with African countries and offer end-to-end nuclear package similar to those used effectively by competitors: coordinated project development, state-backed financing, long-term fuel services, and durable in-country support through regional offices and sustained workforce/regulatory training. With timely planning, sustained political commitment, and appropriate financing and institutional support, nuclear energy, both large reactors and advanced SMRs, can become a meaningful, though not dominant, pillar of Africa’s future power mix, enhancing energy security, enabling industrial growth, and supporting climate goals.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2026

Microreactor Core Transportation Cask Model Description for Criticality Safety Validation Basis Assessment (Rev. 1)

Criticality safety analyses are completed on transportation casks used for microreactor whole core shipment to provide examples of models and analyses to industry, regulators, and nuclear community at large to be used in verification and validation analyses of similar applications. The microreactors considered are based on a Gas-Cooled Microreactor (GCMR) and a Heat-Pipe Microreactor (HPMR), both utilize HALEU fuel in the form of TRISO particles and various other design options considered in industry microreactor designs. Variant design options of GCMR and HPMR were also investigated to provide a wider application range for each technology. Criticality safety analyses for the GCMR and HPMR packages were performed using the CSAS6 sequence of SCALE 6.3.2 with the ENDF/B-VII.1-based continuous energy neutron libraries. Different scenarios were investigated, including normal operation and water flooded conditions to represent nominal and hypothetical accident scenarios. Sensitivity and similarity analyses are also performed using the TSUNAMI sequence of SCALE 6.3.2, and the similarity analysis uses all the experiments from the ICSBEP Handbook with Intermediate and mixed Enriched Uranium (IEU) and Low Enriched Uranium (LEU) systems, together with additional experiments that are sponsored by the DNCSH program. Many experiments were found marginally similar to the GCMR and HPMR models, with similarity index (ck) values greater than 0.8 but less than 0.9. Among all the experiments analyzed, no case has a ck value greater than 0.9, indicating that additional critical experiments might be needed to further validate the criticality safety models for microreactors transport packages.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2025

Improving Dose Modeling With Dynamic Modeling Tools [Slides]

Utilities aiming for higher fuel enrichment for power uprates or extended operation times before refueling must conduct a new dose analysis. Current conservative dose estimation standards may cause utilities to exceed regulatory limits for proposed increased fuel enrichment. A more accurate modeling of doses from reactor accidents can lower these conservative assumptions. Prescott et al. (2022) demonstrated that the Event Modeling Risk Assessment using Linked Diagrams (EMRALD) software tool, developed at Idaho National Laboratory (INL), can be coupled with the Modular Accident Analysis Program (MAAP5) for dynamic accident analysis in reactor plants. EMRALD forms models of potential accident scenarios, while MAAP5 simulates the accident progression and dose consequences. By integrating these software tools with utility-specific data, a more precise estimation of dose consequences from plant accidents can be achieved. Preliminary findings indicate that EMRALD provides accurate mean core damage frequencies for generalized accident scenarios. Future work includes expanding the model to account for plant-specific data and mitigation factors.

97 - MATHEMATICS AND COMPUTING↗
DOE OSTI2025

Post-Irradiation Examination on MiniFuel UCO and UO 2 TRISO Particles Irradiated in HFIR at High Power

Post-irradiation examination (PIE) of MiniFuel compacts was conducted at Oak Ridge National Laboratory (ORNL) under the Nuclear Science User Facilities project in collaboration with Kairos Power (KP) to evaluate the performance of tristructural-isotropic (TRISO) particles under high particle power and fluoride-salt-cooled high-temperature reactor (FHR)-relevant conditions. MiniFuel compacts containing low-enriched uranium oxide-uranium carbide (LEUCO), low-enriched uranium dioxide (LEUO2), and natural UCO (NUCO) kernels were irradiated for four cycles at ORNL’s High Flux Isotope Reactor (HFIR) at target temperatures between 500°C and 900°C. Post irradiation, the experiment was disassembled at ORNL to recover the MiniFuel subcapsules, which were subsequently punctured to measure fission gas release. Subcapsule disassembly allowed the recovery of components of interest, such as silicon carbide (SiC) thermometry, fuel specimens, fission product sinks, and SiC spacers. The experimental irradiation temperature was confirmed by analyzing the SiC thermometry via dilatometry. PIE on the fuel specimens included gamma spectrometry and deconsolidation leach burn leach, which were complemented by imaging techniques such as x-ray computed tomography, optical microscopy, and electron microscopy. The PIE results provide insight into TRISO particle integrity, fission product retention, coating performance, and kernel migration, informing fuel qualification for application in KP’s FHR concept.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2025

Post-Irradiation Examination on MiniFuel UCO and UO 2 TRISO Particles Irradiated in HFIR at High Power

Post-irradiation examination (PIE) of MiniFuel compacts was conducted at Oak Ridge National Laboratory (ORNL) under the Nuclear Science User Facilities project in collaboration with Kairos Power (KP) to evaluate the performance of tristructural-isotropic (TRISO) particles under high particle power and fluoride-salt-cooled high-temperature reactor (FHR)-relevant conditions. MiniFuel compacts containing low-enriched uranium oxide-uranium carbide (LEUCO), low-enriched uranium dioxide (LEUO 2 ), and natural UCO (NUCO) kernels were irradiated for four cycles at ORNL’s High Flux Isotope Reactor (HFIR) at target temperatures between 500°C and 900°C. Post irradiation, the experiment was disassembled at ORNL to recover the MiniFuel subcapsules, which were subsequently punctured to measure fission gas release. Subcapsule disassembly allowed the recovery of components of interest, such as silicon carbide (SiC) thermometry, fuel specimens, fission product sinks, and SiC spacers. The experimental irradiation temperature was confirmed by analyzing the SiC thermometry via dilatometry. PIE on the fuel specimens included gamma spectrometry and deconsolidation leach burn leach, which were complemented by imaging techniques such as x-ray computed tomography, optical microscopy, and electron microscopy. The PIE results provide insight into TRISO particle integrity, fission product retention, coating performance, and kernel migration, informing fuel qualification for application in KP’s FHR concept.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
DOE OSTI2026

Novel Method for Domestic Stable Isotope Production (CRADA #667) Final Report

This report presents the progress achieved on our chlorine isotope separation efforts in the CRADA #667 agreement. We used a microchannel distillation (MCD) packing structure to enrich HCl isotopes (H35Cl, H37Cl) and an isotachophoresis (ITP) separation to enrich ionic chloride species (35Cl- and 37Cl-). A 2 m tall MCD column was constructed and achieved 264 separation stages over several days, which is well above the goal of 50 stages outlined in the CRADA. The resulting height equivalent to a theoretical plate (HETP) was 0.76 cm. ITP tests were run ~ 24 hours and achieved a single-stage separation factor of 1.559.

07 ISOTOPE AND RADIATION SOURCES↗
DOE OSTI2025

Inter-Kingdom Viral Interactions

Please cite as : Josué A. Rodríguez-Ramos, Amy E. Zimmerman, Ruonan Wu, Sheryl Bell, Trinidad Alfaro, Kirsten Hofmockel, William C. Nelson. 2025. Inter-Kingdom Viral Interactions. [Data Set] PNNL DataHub. This data is published under a CC0 license. The authors encourage data reuse and request attribution by referencing the above citations for the data package and associated manuscript. Deciphering viral ecology in soils is challenging due to their high physiochemical and community complexity. To enhance detection of sub-communities of DNA and RNA viruses, we applied fractionation approaches to soils collected across a moisture gradient from a grassland field experiment. Analyses included metagenomics and metatranscriptomics of size-fractionated extracellular viruses (i.e., DNA and RNA viromes), metagenomics of bacteria/archaea- or eukaryote-enriched samples, and whole soil metatranscriptomes with rRNA-depletion or polyadenylation enrichment. While RNA virome and whole soil RNA methods captured similar viral diversity, RNA viromes identified longer, higher-quality genomes. Further, we showed that significantly more DNA viruses were active in higher moisture than lower moisture samples, whereas responses by overall diversity vary by genome type (DNA versus RNA genomes). Finally, we demonstrate the power of fractionation approaches for identifying distinct viral communities that infect unique hosts, which has significant implications for ecological investigations, particularly related to interkingdom interactions.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

WA-IsoC_MSC1.1.0

The soil microbiome is central to the cycling of carbon and other nutrients and to the promotion of plant growth. Despite its importance, analysis of the soil microbiome is difficult due to its sheer complexity, with thousands of interacting species. Here, we reduced this complexity by developing model soil microbial consortia that are simpler and more amenable to experimental analysis but still represent important microbial functions of the native soil ecosystem. Samples were collected from an arid grassland soil and microbial communities (consisting mainly of bacterial species) were enriched on agar plates containing chitin as the main carbon source. Chitin was chosen because it is an abundant carbon and nitrogen polymer in soil that often requires the coordinated action of several microorganisms for complete metabolic degradation. Several soil consortia were derived that had tractable richness (30-50 OTUs) with diverse phyla representative of the native soil, including Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria and Verrucomicrobia. The resulting consortia can be stored as glycerol or lyophilized stocks at -80° C and revived while retaining community composition, greatly increasing their use as tools for the research community at large. One of the consortia that was particularly stable was chosen as a model soil consortium (MSC-1) for further analysis. MSC-1 species interactions were studied using both pairwise co-cultivation in liquid media and during growth in soil under several perturbations. Co-abundance analyses highlighted interspecies interactions and helped to define keystone species, including Mycobacterium, Rhodococcus, and Rhizobiales taxa. These experiments demonstrate the success of an approach based on naturally enriching a community of interacting species that can be stored, revived, and shared. The knowledge gained from querying these communities and their interactions will enable better understanding of the soil microbiome and the roles these interactions play in this environment.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2024

KBase Narrative - van den Bergh et al

We report 14 metagenome-assembled genomes (MAGs) recovered from agricultural soils, composts, and enrichment cultures enriched therefrom, originating from the Netherlands, Belgium and Switzerland. Recovered MAGs constitute methane-cycling related organisms sequenced from isolated cultures or environmental DNA, elucidating their metabolic potential in important ecological processes like the oxidation or production of atmospheric trace gases as methane, hydrogen, and carbon monoxide.

Rocha, Gabriel Silvestre↗
DOE OSTI2021

ACCRUE—An Integral Index for Measuring Experimental Relevance in Support of Neutronic Model Validation

A key challenge for the introduction of any design changes, e.g., advanced fuel concepts, first-of-a-kind nuclear reactor designs, etc., is the cost of the associated experiments, which are required by law to validate the use of computer models for the various stages, starting from conceptual design, to deployment, licensing, operation, and safety. To achieve that, a criterion is needed to decide on whether a given experiment, past or planned, is relevant to the application of interest. This allows the analyst to select the best experiments for the given application leading to the highest measures of confidence for the computer model predictions. The state-of-the-art methods rely on the concept of similarity or representativity, which is a linear Gaussian-based inner-product metric measuring the angle—as weighted by a prior model parameters covariance matrix—between two gradients, one representing the application and the other a single validation experiment. This manuscript emphasizes the concept of experimental relevance which extends the basic similarity index to account for the value accrued from past experiments and the associated experimental uncertainties, both currently missing from the extant similarity methods. Accounting for multiple experiments is key to the overall experimental cost reduction by prescreening for redundant information from multiple equally-relevant experiments as measured by the basic similarity index. Accounting for experimental uncertainties is also important as it allows one to select between two different experimental setups, thus providing for a quantitative basis for sensor selection and optimization. The proposed metric is denoted by ACCRUE, short for Accumulative Correlation Coefficient for Relevance of Uncertainties in Experimental validation. Using a number of criticality experiments for highly enriched fast metal systems and low enriched thermal compound systems with accident tolerant fuel concept, the manuscript will compare the performance of the ACCRUE and basic similarity indices for prioritizing the relevance of a group of experiments to the given application.

97 MATHEMATICS AND COMPUTING↗
DOE OSTI2020

Metagenomics Reveals Diet-Specific Specialization of Bacterial Communities in Fungus Gardens of Grass- and Dicot-Cutter Ants

Leaf-cutter ants in the genus Atta are dominant herbivores in the Neotropics. While most species of Atta cut dicots to incorporate into their fungus gardens, some species specialize on grasses. Here we examine the bacterial community associated with the fungus gardens of grass- and dicot-cutter ants to examine how changes in substrate input affect the bacterial community. We sequenced the metagenomes of 12 Atta fungus gardens, across four species of ants, with a total of 5.316 Gbp of sequence data. We show significant differences in the fungus garden bacterial community composition between dicot- and grass-cutter ants, with grass-cutter ants having lower diversity. Reflecting this difference in community composition, the bacterial functional profiles between the fungus gardens are significantly different. Specifically, grass-cutter ant fungus garden metagenomes are particularly enriched for genes responsible for amino acid, siderophore, and terpenoid biosynthesis while dicot-cutter ant fungus gardens metagenomes are enriched in genes involved in membrane transport. Differences between community composition and functional capacity of the bacteria in the two types of fungus gardens reflect differences in the substrates that the ants incorporated. These results show that different substrate inputs matter for fungus garden bacteria and shed light on the potential role of bacteria in mediating the ants’ transition to the use of a novel substrate.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Metagenomic and Metatranscriptomic Analyses Revealed Uncultured Bacteroidales Populations as the Dominant Proteolytic Amino Acid Degraders in Anaerobic Digesters

Current understanding of amino acid (AA) degraders in anaerobic digesters is mainly based on cultured species, whereas microorganisms that play important roles in a complex microbial community remain poorly characterized. This study investigated short-term enrichments degrading single AAs using metagenomics and metatranscriptomics. Metagenomic analysis revealed that populations related to cultured AA degraders had an abundance <2.5% of the sequences. In contrast, metagenomic-assembled bins related to uncultured Bacteroidales collectively accounted for >35% of the sequences. Phylogenetic analyses suggested that these Bacteroidales populations represented a yet-to-be characterized family lineage, i.e., Bacteroidetes vadinHA17. The bins possessed the genetic capacity related to protein degradation, including surface adhesion (3–7 genes), secreted peptidase (52–77 genes), and polypeptide-specific transporters (2–5 genes). Furthermore, metatranscriptomics revealed that these Bacteroidales populations expressed the complete metabolic pathways for degrading 16 to 17 types of AAs in enrichments fed with respective substrates. These characteristics were distinct from cultured AA degraders including Acidaminobacter and Peptoclostridium, suggesting the uncultured Bacteroidales were the major protein-hydrolyzing and AA-degrading populations. These uncultured Bacteroidales were further found to be dominant and active in full-scale anaerobic digesters, indicating their important ecological roles in the native habitats. “Candidatus Aminobacteroidaceae” was proposed to represent the previously uncharted family Bacteroidetes vadinHA17.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Microbial Community Field Surveys Reveal Abundant Pseudomonas Population in Sorghum Rhizosphere Composed of Many Closely Related Phylotypes

While the root-associated microbiome is typically less diverse than the surrounding soil due to both plant selection and microbial competition for plant derived resources, it typically retains considerable complexity, harboring many hundreds of distinct bacterial species. Here, we report a time-dependent deviation from this trend in the rhizospheres of field grown sorghum. In this study, 16S rRNA amplicon sequencing was used to determine the impact of nitrogen fertilization on the development of the root-associated microbiomes of 10 sorghum genotypes grown in eastern Nebraska. We observed that early rhizosphere samples exhibit a significant reduction in overall diversity due to a high abundance of the bacterial genus Pseudomonas that occurred independent of host genotype in both high and low nitrogen fields and was not observed in the surrounding soil or associated root endosphere samples. When clustered at 97% identity, nearly all the Pseudomonas reads in this dataset were assigned to a single operational taxonomic unit (OTU); however, exact sequence variant (ESV)-level resolution demonstrated that this population comprised a large number of distinct Pseudomonas lineages. Furthermore, single-molecule long-read sequencing enabled high-resolution taxonomic profiling revealing further heterogeneity in the Pseudomonas lineages that was further confirmed using shotgun metagenomic sequencing. Finally, field soil enriched with specific carbon compounds recapitulated the increase in Pseudomonas, suggesting a possible connection between the enrichment of these Pseudomonas species and a plant-driven exudate profile.

59 BASIC BIOLOGICAL SCIENCES↗