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282 records · Page 16
All sourcesNASA NTRSDOE OSTI
DOE OSTI2020

Biotic Interactions Are More Important than Propagule Pressure in Microbial Community Invasions

ABSTRACT Microbial probiotics are intended to improve functions in diverse ecosystems, yet probiotics often fail to establish in a preexisting microbiome. This is a species invasion problem. The relative importance of the two major factors controlling establishment in this context—propagule pressure (inoculation dose and frequency) and biotic interactions (composition of introduced and resident communities)—is unknown. We tested the effect of these factors in driving microbial composition and functioning following 12 microbial community invasions (e.g., introductions of many microbial invaders) in microcosms. Ecosystem functioning over a 30-day postinvasion period was assessed by measuring activity (respiration) and environment modification (dissolved organic carbon abundance). To test the dependence on environmental context, experiments were performed in two resource environments. In both environments, biotic interactions were more important than propagule pressure in driving microbial composition and community function, but the magnitude of effect varied by environment. Successful invaders comprised approximately 8% of the total number of operational taxonomic units (OTUs). Bacteria were better invaders than fungi, with average relative abundances of 7.4% ± 6.8% and 1.5% ± 1.4% of OTUs, respectively. Common bacterial invaders were associated with stress response traits. The most resilient bacterial and fungal families, in other words, those least impacted by invasions, were linked to antimicrobial resistance or production traits. Illuminating the principles that determine community composition and functioning following microbial invasions is key to efficient community engineering. IMPORTANCE With increasing frequency, humans are introducing new microbes into preexisting microbiomes to alter functioning. Example applications include modification of microflora in human guts for better health and those of soil for food security and/or climate management. Probiotic applications are often approached as trial-and-error endeavors and have mixed outcomes. We propose that increased success in microbiome engineering may be achieved with a better understanding of microbial invasions. We conducted a microbial community invasion experiment to test the relative importance of propagule pressure and biotic interactions in driving microbial community composition and ecosystem functioning in microcosms. We found that biotic interactions were more important than propagule pressure in determining the impact of microbial invasions. Furthermore, the principles for community engineering vary among organismal groups (bacteria versus fungi).

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Allosteric control of electron transfer in nitrogenase and nitrogenase-like enzymes (Final Report)

More than a third of all known proteins to date are either oxidoreductases or metalloproteins. The diversity of metal cofactors and redox centers in these proteins make them attractive candidates for engineering protein-based biocatalysts, biosensors, and biofuel cells, among other necessary tools for the bioeconomy. Many of these enzymes function as large multi-subunit complexes arranged as symmetric or asymmetric assemblies, and the purpose of this architecture is unclear. The goal of the project was to address this fundamental question by investigating the mechanism of action of two oxidoreductases: Nitrogenase and DPOR (Dark-Operative Protochlorophyllide Oxidoreductase, a nitrogenase-like oxidoreductase). Nitrogenase and DPOR play key roles in nitrogen fixation and photosynthesis, processes essential to sustaining life. Both proteins are arranged as two symmetrical halves with each half capable of substrate reduction. For more than half a century, substrate reduction by nitrogenase has been presumed to occur independently within the two identical halves. Funded by this DOE grant we have now shown that both nitrogenase and DPOR function asymmetrically. Electron transfer in one half suppresses activity in the other. Thus, we have demonstrated that the two halves do not function in an independent manner; rather, catalytic events in one-half allosterically suppress activity and electron transfer (ET) in the other. This alternating-site mechanism has significant implications on our current understanding of substrate reduction by these enzymes and offers a new mechanistic paradigm for the reactions they catalyze. Our work now provides a mechanistic map of the how the two symmetric catalytic halves work together to establish and maintain the sequential-site mechanism through multiple ET cycles to achieve substrate reduction. There is substantive need for bio-inspired tools that enable reduction of nitrogen and other substrates, as well as photosynthesis, for energy and food production. Understanding the workings of enzymes like nitrogenase and DPOR that mediate multi-electron redox chemistry is vital for designing such synthetic tools. The timing of our research coincides with the biotechnological breakthrough of light-driven artificial N2 reduction by nitrogenase, achieved using the native enzyme with both halves arranged in a mirror-like architecture as catalyst. Together with such discoveries, our research now reveal unknown facets of the reaction mechanism and hold promise for designing more effective catalysts.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Exploring small-scale chemostats to scale up microbial processes: 3-hydroxypropionic acid production in S. cerevisiae

The physiological characterization of microorganisms provides valuable information for bioprocess development. Chemostat cultivations are a powerful tool for this purpose, as they allow defined changes to one single parameter at a time, which is most commonly the growth rate. The subsequent establishment of a steady state then permits constant variables enabling the acquisition of reproducible data sets for comparing microbial performance under different conditions. We performed physiological characterizations of a 3-hydroxypropionic acid (3-HP) producing Saccharomyces cerevisiae strain in a miniaturized and parallelized chemostat cultivation system. The physiological conditions under investigation were various growth rates controlled by different nutrient limitations (C, N, P). Based on the cultivation parameters obtained subsequent fed-batch cultivations were designed.We report technical advancements of a small-scale chemostat cultivation system and its applicability for reliable strain screening under different physiological conditions, i.e. varying dilution rates and different substrate limitations (C, N, P). Exploring the performance of an engineered 3-HP producing S. cerevisiae strain under carbon-limiting conditions revealed the highest 3-HP yields per substrate and biomass of 16.6 %C-mol and 0.43 g gCDW -1 , respectively, at the lowest set dilution rate of 0.04 h -1 . 3-HP production was further optimized by applying N- and P-limiting conditions, which resulted in a further increase in 3-HP yields revealing values of 21.1 %C-mol and 0.50 g gCDW -1 under phosphate-limiting conditions. The corresponding parameters favoring an increased 3-HP production, i.e. dilution rate as well as C- and P-limiting conditions, were transferred from the small-scale chemostat cultivation system to 1-L bench-top fermenters operating in fed-batch conditions, revealing 3-HP yields of 15.9 %C-mol and 0.45 g gCDW -1 under C-limiting, as well as 25.6 %C-mol and 0.50 g gCDW -1 under phosphate-limiting conditions. Small-scale chemostat cultures are well suited for the physiological characterization of microorganisms, particularly for investigating the effect of changing cultivation parameters on microbial performance. In our study, optimal conditions for 3-HP production comprised (i) a low dilution rate of 0.04 h -1 under carbon-limiting conditions and (ii) the use of phosphate-limiting conditions. Similar 3-HP yields were achieved in chemostat and fed-batch cultures under both C- and P-limiting conditions proving the growth rate as robust parameter for process transfer and thus the small-scale chemostat system as powerful tool for process optimization.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Plant Growth Study: Growth of Three Plant Species in 100-OL-1 Operable Unit Soils

Soil contamination from historical application of lead arsenate pesticides persists in the 100-OL-1 Operable Unit (OU) of the U.S. Department of Energy’s Hanford Site in Benton County, Washington. A remedial investigation used a portable X-ray fluorescence analyzer to estimate and map the concentrations of metals in surface soil in the 133 decision units comprising the OU. There is variability in lead and arsenic concentrations among the non-contiguous decision units, and an incomplete understanding of the ecological and human risks from soil contamination. While general screening criteria for lead are reported in literature, limited information is available on the impacts of heavy metals on site-specific plant and animal life. To address this information gap, a study on plant growth in Hanford soils was conducted using native bluegrass (Poa secunda), invasive cheatgrass (Bromus tectorum), and buttercrunch lettuce (Lactuca sativa). The latter is included because it is a common species used in plant growth studies. Soil samples were collected from a single general location within the 100-OL-1 OU, prepared, and categorized as having low, medium, or high concentrations of lead and arsenic, with the high concentration soil having concentrations of up to 3,400 and 790 mg/kg lead and arsenic, respectively. Additionally, a synthetic soil was prepared and used as a control, in accordance with standard plant growth protocols. Fifty-four seeds of each plant species were planted in each of the four soil types following the procedure outlined by the Washington State Department of Ecology. Germination rates and biomass measurements were recorded for 20 days. Both bluegrass and lettuce germination rates appeared to be reduced at the highest lead and arsenic concentrations (Pb: 3400 mg/kg, As: 790 mg/kg), while cheatgrass germination rates were unaffected. Total biomass for all species appeared to be related to the relative concentrations of lead and arsenic in the soil. Results of this growth study agree reasonably well with previous Hanford Site plant growth studies and provide additional data for assessing ecological risk at the Hanford Site.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

Design Study for Bio-Oil Production from Biomass Using a Dual Fluidized-Bed Reactor

To evaluate the re-design and reconfiguration of a dual-fluidized bed (DFB) gasification system into a recirculating pyrolysis reactor, Computation Fluid Dynamic (CFD) simulations of the system were conducted. The Barracuda Virtual Reactor® computational particle fluid dynamic code was used to perform simulations of the pyrolysis process. Modeling of the chemical reaction kinetics for both gas phase and solid particle phase were included. The recirculating pyrolysis reactor shown in Fig. 1a is based on a bubbling-bed biomass pyrolyzer and a riser combustor to convert the remaining char. The operational differences in the re-configuration of the DFB gasification system into a recirculating pyrolysis system for the production of bio-oil are (1) replacement of a low-surface area inert bed material with a high-surface-area bed material that has acidic properties to provide catalytic activity for the production of bio-oil with reduced oxygen content, (2) lower temperature and residence time for bio-oil production from pyrolysis, (3) replacement of the fluidization gas in the bubbling bed pyrolyzer from steam to nitrogen, and (4) the reduction of pyrolyzer freeboard volume. The bed material used for catalytic pyrolysis is Sasol 300 (300-micron dia., bulk density 0.94 kg/l, and surface area 130 m2/g) and is a theta-alumina with mild acidity. This is in comparison with previous standard bed material Carbo HSP (430-micron dia., bulk density 2.01 kg/l, and surface area 0.03 m2/g) used for gasification. For bio-oil production, pyrolysis in the bubbling bed requires temperatures in the range of 550 C in comparison with gasification temperatures near 850 C. To attain this lower temperature requires management of the energy mass balances, with control of the bed material recirculation rate between bubbling bed pyrolyzer and riser combustor, the introduction of a nitrogen purge in the pyrolyzer, and adjusting the pressure balance between the two vessels. To extract bio-oil from the pyrolysis reactor with a snorkel, two different freeboard configurations were evaluated. In Fig. 1 b the existing high freeboard configuration is shown and in Fig. 1 c the reduced freeboard design is presented. The introduction of a nitrogen purge for the high free board configuration provided the highest bio-oil production from the CFD simulations. To decrease the bed-material circulation rate, primary and secondary air on the combustor side were reduced, and the pressure on the combustor was slightly increased. A portion of the biomass and bio-oil was observed to be transported to the combustor, leading to a smaller pyrolysis yield. The control of the pyrolyzer temperature is performed by controlling the circulation rate. Good fluidization of the bubbling bed and cascade PID control are required to keep the temperature from oscillating due to large time delay experienced when changing primary and secondary air. (a) (b) (c) Figure 1. (a) Dual Fluidized Bed pyrolysis system configuration, (b) mole fraction of nonpolar biooil in high freeboard configuration, and (c) mole fraction of nonpolar bio-oil in low freeboard configuration with nitrogen purge introduced in both configurations.

BASIC BIOLOGICAL SCIENCES,BIOMASS FUELS↗
DOE OSTI2021

The phylogenetic roots of addiction: compulsive drug seeking, natural and drug-sensitive reward, and the acquisition of learned habits

Our rational faculties permit us humans to maximize the utility of our actions. We perform a fundamental type of cost-benefit analysis in which we frame a problem, assign values to the different paths, and then choose from among a set of available options, the course that promises the most favorable outcome. So why then does addiction appear to be so impervious to the associated costs, so unaffected by undesired consequences, and ultimately so resistant to cognitive oversight? The answer may likely be found in the fact that the drivers for compulsive drug seeking and drug taking are located in affective brain circuits, circuits that are structured and patterned by learning with repeated activation. These deep processes exhibit significant resistance to control by our cognitive faculties. The general consensus is that addiction arises from mechanisms that overvalue the magnitude of reward, discount the associated risks, and thereby bias individuals towards compulsive pursuit of addictive drugs. Behavioral disruption and dependence appear to arise at the intersection of a number of connected but separate phenomena: expectations for the occurrence of specific events, behaviors that seek encounters with them, the ability to notice and learn nonrandom, co-occurring conditions, the prediction and valuation of consequences, the forming of enduring memories, and the drivers of focused behavior through compulsion, habits, and acquired routines. It is important to recognize that each one of these individual faculties are present and well developed across the entire phylogenetic tree of bilateral metazoans. The goal of this special volume is dedicated to exploring the degree to which inherent elements can account for addiction and addiction-associated phenomena. In much of the literature on addiction, the underlying processes are often viewed as distinctly mammalian, arising, in part, from the strong cognitive capacities of this taxon. Some phenomena may even be regarded to exist only in primates, or even solely in humans. This supposition arises from the fact that studies are conducted almost exclusively in mammals and primates, while evolutionary antecedents of the behavior are rarely considered. A more comprehensive perspective that examines drug reward and reinforcement in a wider range of organisms demonstrates that many of the component traits are actually well developed across the greater metazoan lineage, and they may well predate the emergence of a mammalian clade by a wide margin. The collection of papers assembled here supports the notion that the capacity to associate cues and quences has not arisen in mammals de novo. Rather, the neural mechanisms for detecting contingencies and for predicting future outcomes are very deeply rooted across broad phylogenetic divisions. Our understanding of an ability to associate paired events has been enriched by work in invertebrate preparations in both classical and operant conditioning scenarios [Cook and Carew, 1986, 1989a, 1989b]. The ability to learn allows us to connect cues and behavioral actions to their associated consequences. Pavlovian conditioning enriches surrounding cues with predictive value. Outcomes with positive valence generate appetitive responses and approach to the associated cues, while those perceived as aversive bring cue avoidance and withdrawal. Humans are not the only life forms capable of such short- and long-term modulations of behavior

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2016

Remote Dismantling and Packaging of the RPV and Thermal Shield at the Obrigheim NPP - 16073

The Obrigheim Nuclear Power Plant (KWO) is located in the Federal State of Baden- Wuerttemberg in Germany and was operated by the EnKK-KWO. The Nuclear Power Plant - a light water pressurized water reactor with 357 MW of electrical power - was operated from 1968 to 2005. In June 2009 the Energiewerke Nord GmbH (EWN) was charged with the dismantling planning and dismantling of the reactor. According to the German nuclear and radiation protection law, several licenses will be required for the realisation of this task. This also applies to the disposal of the waste produced by dismantling, which must finally be stored in the German final repository for low and medium-radioactive waste (underground repository 'Konrad'). In spring 2010 EWN completed all application documents for the dismantling of the reactor pressure vessel (RPV) and the RPV internals and EnKK-KWO delivered them to the responsible Environmental Ministry of Baden-Wuerttemberg. Since then it has been worked intensively on the design, construction and manufacturing of the facilities for dismantling. With a separate notification of amendment, it was applied for the license to install and commission the facilities for the dismantling of the RPV and the RPV internals at the Environmental Ministry of Baden-Wuerttemberg in October 2010. This notification of amendment was confirmed in summer 2012. In September 2011 the work process and test plan and the technical information about the campaign for the removal of the radioactive waste resulting from the RPV with internals, peripheral components and secondary waste with the aim of final disposal in the Federal Repository 'Konrad' was submitted to the Federal Office for Radiation Protection. The evaluation and assessment of the work process and test plan was concluded in August 2013, when it was audited by TUV Nord EnSys without any further measures and indications. On September 10 the work process and test plan was released by the Federal Office for radiation protection. The dismantling of the RPV internals upper core and lower core structure started in September 2013 in the wet cutting area in room 01.106/306 and were finished in October 2014. In the presentation 'Dismantling of the Obrigheim NPP Reactor and Waste Management' held on the WM Symposia in Phoenix, 2014, an overview about the remote cutting and packaging of the upper and lower core structure was given. In today's presentation the subsequent dismantling activities will be outlined and evaluated. The presentation will start with the short description of the basic frame conditions for the nuclear licensing procedure, important procedural principles, and necessary plant-specific conditions. After the remote cutting and packing of the reactor internals of the upper and lower core structure was completed, the wet cutting area 'reactor room 01.106/306' was cleaned and the reactor room rebuilt into a cutting area for peripheral components. After the peripheral components were dismantled, all preconditions were set to lift the RPV from its mounting position and to arrange it in the upper reactor room 01.306. After these activities were completed, the lifting of the RPV was prepared and the cutting area 'Dismantling of the RPV insulation in the reactor room 01.106/306' was set up. To carry out this task, a strand jack unit with connection block was installed on the bridge of the reactor building crane. The opening created by the dismantling of the pool plate was closed with a flap door. The manipulators used for the wet cutting of the RPV internals were also used for the remote dismantling of the RPV insulation and mounted in the cutting area. The remote facilities were handled from a control centre and/or control stand. The air-technical housing which has already been used for the remote dismantling of the RPV internals was designed for the dismantling of the RPV insulation and was extended for this dismantling scope with appropriate ventilation systems. A special feature is the fixation of the RPV insulation at the RPV corpus. The RPV insulation consists of insulation wool, wire mesh and metal-sheet insulation. This 'Sandwich Construction' is detached with a steel corset at the RPV corpus and partly tightened. The insulation of the RPV bottom dome is tightened at the supporting frame. Due to the activation of the material in the core area and the resulting high ambient dose rate the RPV insulation can only be dismantled remotely. Additionally, as asbestos fibers are expected to be found in the insulation the dismantling has to be performed according to the Technical Rules for hazardous substances. The demolition, restoration or maintenance works have to be performed remotely according to the Technical Rules for hazardous substances when the RPV is located in the cutting area. But in case of dismantling of a power reactor no such experience can be found. Beginning with the explanation of the technical, radiological and substantial boundary conditions this presentation will not only impart knowledge about the dismantling of the RPV insulation but also about the remote controlled packaging of the metal sheet insulation and the manual conditioning of the RPV insulation. Subsequently, reports on the remotely controlled cutting and packaging of the RDB are given. Here is particularly interesting that cutting methods have been used and how the activity determination of the segments was carried out. On the basis of the evaluation of the realisation the subsequent dismantling activities will be lined out. In parallel to the dismantling tasks, the waste package documentation for the final repository was issued, checked and confirmed for the containers loaded with parts of the RDB internals. The most important stages are exemplified in this presentation. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2018

Soil Viruses Are Underexplored Players in Ecosystem Carbon Processing

ABSTRACT Rapidly thawing permafrost harbors ∼30 to 50% of global soil carbon, and the fate of this carbon remains unknown. Microorganisms will play a central role in its fate, and their viruses could modulate that impact via induced mortality and metabolic controls. Because of the challenges of recovering viruses from soils, little is known about soil viruses or their role(s) in microbial biogeochemical cycling. Here, we describe 53 viral populations (viral operational taxonomic units [vOTUs]) recovered from seven quantitatively derived (i.e., not multiple-displacement-amplified) viral-particle metagenomes (viromes) along a permafrost thaw gradient at the Stordalen Mire field site in northern Sweden. Only 15% of these vOTUs had genetic similarity to publicly available viruses in the RefSeq database, and ∼30% of the genes could be annotated, supporting the concept of soils as reservoirs of substantial undescribed viral genetic diversity. The vOTUs exhibited distinct ecology, with different distributions along the thaw gradient habitats, and a shift from soil-virus-like assemblages in the dry palsas to aquatic-virus-like assemblages in the inundated fen. Seventeen vOTUs were linked to microbial hosts ( in silico ), implicating viruses in infecting abundant microbial lineages from Acidobacteria , Verrucomicrobia , and Deltaproteobacteria , including those encoding key biogeochemical functions such as organic matter degradation. Thirty auxiliary metabolic genes (AMGs) were identified and suggested virus-mediated modulation of central carbon metabolism, soil organic matter degradation, polysaccharide binding, and regulation of sporulation. Together, these findings suggest that these soil viruses have distinct ecology, impact host-mediated biogeochemistry, and likely impact ecosystem function in the rapidly changing Arctic. IMPORTANCE This work is part of a 10-year project to examine thawing permafrost peatlands and is the first virome-particle-based approach to characterize viruses in these systems. This method yielded >2-fold-more viral populations (vOTUs) per gigabase of metagenome than vOTUs derived from bulk-soil metagenomes from the same site (J. B. Emerson, S. Roux, J. R. Brum, B. Bolduc, et al., Nat Microbiol 3:870–880, 2018, https://doi.org/10.1038/s41564-018-0190-y). We compared the ecology of the recovered vOTUs along a permafrost thaw gradient and found (i) habitat specificity, (ii) a shift in viral community identity from soil-like to aquatic-like viruses, (iii) infection of dominant microbial hosts, and (iv) carriage of host metabolic genes. These vOTUs can impact ecosystem carbon processing via top-down (inferred from lysing dominant microbial hosts) and bottom-up (inferred from carriage of auxiliary metabolic genes) controls. This work serves as a foundation which future studies can build upon to increase our understanding of the soil virosphere and how viruses affect soil ecosystem services.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2017

Endomembrane Cation Transporters and Membrane Trafficking

Multicellular, as well as unicellular, organisms have evolved mechanisms to regulate ion and pH homeostasis in response to developmental cues and to a changing environment. The working hypothesis is that the balance of fluxes mediated by diverse transporters at the plasma membrane and in subcellular organelles determines ionic cellular distribution, which is critical for maintenance of membrane potential, pH control, osmolality, transport of nutrients, and protein activity. An emerging theme in plant cell biology is that cells respond and adapt to diverse cues through changes of the dynamic endomembrane system. Yet we know very little about the transporters that might influence the operation of the secretory system in plants. Here we focus on transporters that influence alkali cation and pH homeostasis, mainly in the endomembrane/ secretory system. The endomembrane system of eukaryote cells serves several major functions: i) sort cargo (e.g. enzymes, transporters or receptors) to specific destinations, ii) modulate the protein and lipid composition of membrane domains through remodeling, and iii) determine and alter the properties of the cell wall through synthesis and remodeling. We had uncovered a novel family of predicted cation/H + exchangers (CHX) and K + efflux antiporters (KEA) that are prevalent in higher plants, but rare in metazoans. We combined phylogenetic and transcriptomic analyses with molecular genetic, cell biological and biochemical studies, and have published the first reports on functions of plant CHXs and KEAs. CHX studied to date act at the endomembrane system where their actions are distinct from the better-studied NHX (Na/K-H + exchangers). Arabidopsis thaliana CHX20 in guard cells modulate stomatal opening, and thus is significant for vegetative survival. Other CHXs ensure reproductive success on dry land, as they participate in organizing pollen walls, targeting of pollen tubes to the ovule or promoting fertilization. Based on localization and mutant analyses, we conclude that CHXs modulate the ion balance, pH or both in micro-regions of endoplasmic reticulum, endosomes and prevacuolar compartment (PVC), and so influence membrane trafficking and signaling resulting in proper osmoregulation in guard cells and seed formation. We also demonstrated for the first time that AtKEA2 associates with chloroplasts, especially at the two poles of developing plastids. These results show that AtKEA1 and AtKEA2 transporters in specific microdomains of the inner envelope link local osmotic, ionic, and pH homeostasis to plastid division and thylakoid membrane formation. The first 3-D structure model of AtCHX was generated, and architecture-directed mutagenesis identified critical residues of the transport core giving insights to the transport mode of this family. Thus we have revealed for the first time crucial roles of an unknown K + /H + transport family on plant growth (KEA), gas exchange, pollen cell wall, and different phases of reproduction (CHXs). The dynamic endomembrane of plant cells is integral to cytokinesis, cell expansion, defense, and cell wall formation, thus these studies are directly relevant to the mission of the Department of Energy and to a better understanding of determinants for enhancing plant biomass and plant tolerance to abiotic stress.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Technical Performance and Cost Optimization of Unobtrusive Multi-static Serial LiDAR Imager (UMSLI) for Wide-area Surveillance and Identification of Marine Life at Marine Energy Installations

Florida Atlantic University developed an underwater optical monitoring system prototype - Unobtrusive Multi-static Serial LiDAR Imager (UMSLI) - suitable for marine energy full project lifecycle observation (baseline, commissioning, and decommissioning), with an automated real-time classification of marine animals. With precursor 2014 DOE funding (award DE-EE0006787), a prototype UMSLI was demonstrated in a controlled laboratory environment and achieved a TRL 6. This EERE DE-EE0007828 award aimed to both increase the TRL of the UMSLI by improving the technology performance (e.g., increase distance of marine animal target detection capability, add additional species classification capabilities, improve the system performance during the day, etc.) and reduce the cost. The UMSLI presents a novel application of underwater distributed Light Detection And Ranging (LiDAR), an advanced remote sensing method that uses light in the form of laser pulses for an application, this paired with an algorithm provides 360 degrees underwater detection, imaging, and classification of marine life. This solution for underwater monitoring of biota preserves the advantages of traditional optical and acoustic solutions while overcoming many associated disadvantages for marine energy site environmental monitoring, such as difficulties in species detection and classification in low light or night and turbid environments. This new approach is a purposefully designed, reconfigurable adaptation of an existing class 3B laser technology into one UMSLI instrument that can be easily mounted on or around different classes of marine energy equipment, such as devices to capture ocean current energy or wave energy. The system uses relatively low average power, utilizes far-red (> 635nm) laser illumination to be invisible and eye-safe to marine animals, is compact, and cost-effective. The equipment is designed for long-term, maintenance-free operations (i.e., current design is targeting more than 7 days continuous operation), to inherently generate a sparse primary dataset that only includes detected anomalies, and to allow robust real-time automated animal classification and identification with a low data bandwidth requirement. The technology’s overarching goal for application, is a system that can be deployed to collect pre-installation baseline species observations at a proposed marine energy deployment site with minimal post-processing overhead. The envisioned system will also produce high-resolution imagery of marine animals through a wide range of conditions and support automated tracking and notification of the presence of managed animals within established perimeters of marine energy equipment to satisfy deployed marine energy projects’ endangered and threatened species monitoring requirements. Through the current project, we demonstrated the UMSLI prototype in an operational environment and increased the UMSLI’s Technology Readiness Level from 6 to 7. The project resulted in many novel technologies, including: 1) an eye-safe red laser based, low-cost LiDAR system that can detect targets up to 10 meters distance; 2) the GAN-based machine learning underwater LiDAR image enhancement technique (this is the first known application of GAN technique in underwater LiDAR); 3) LiDAR-based real-time automated detection capabilities; and 4) a template matching based automated classification tool. These technologies build a solid foundation for future efforts to develop an extended range electro-optical monitoring system suitable for marine energy deployments. In addition to technology development, a key lesson learned is that addressing regulatory and safety requirements must be front and center in any marine energy monitoring applications.

16 TIDAL AND WAVE POWER↗
DOE OSTI2019

Remote Technology to Retrieve Waste Below the 324 Building at the Hanford Site - 19622

The Waste Technology Engineering Laboratory (324 Building) is a Category 2 Nuclear Facility at Hanford. The 324 Building Disposition Project is responsible for the removal and remedial actions under CERCLA. Additionally, there are RCRA Dangerous Waste Management Units which the CERCLA actions are presumed to satisfy. The Radiochemical Engineering Complex (REC) is a group of four hot cells (A, B, C and D Cells) with an interconnecting airlock inside of the 324 Building. Underneath the REC B-Cell are contaminated soils that have an estimated 12,793 R/Hr dose rate and remediation is under CERCLA 300-FF-2 Operable Unit. The current action consists of debris removal from the floor of B-Cell, B-Cell floor removal, and excavation of the soils beneath the floor to a depth of up to twelve feet with four Remote Excavator Arms (REA). This paper provides insight into the preparation and performance of high tolerance core drilling into a hot cell complex. Installation of the REA Through Support (REA TS) mechanism through the 4'-6' concrete shield walls is a complex series of tasks with demanding tolerances at each of four locations around the perimeter of B-Cell. The use of training mock ups was included in the completion of this work. The REA TS is two 11 inch diameter high strength steel sleeves that have expandable collars at each end allowing the interstitial space between the outside of the sleeve and inside of the core drill to be grouted to form a substantial structural connection, and a 10 inch pipe with hydraulic hoses to power the REA. There are three cores required for each REA location. The REAs themselves are customized excavator arms, mounted on support columns to be used in the excavation and transfer of the contaminated soil into storage facilities. The tasks for the installation of the REA TSs are: 1) Characterization of B-Cell Cubicles. 2) Embedded Pipe Fill. 3) Interference Removal. a. Internal Walls, b. External Walls. 4) Core Drilling. 5) Go/No-Go gauge, and video inspection. 6) Installation of REA Through Support Assembly. The 324 Building REC Hot Cells have internal, external, and embedded utilities throughout the shield walls in conflict with the REA TS locations. Some of the embedded utilities have connection points inside of the B-Cell Cubicles. There were two separate issues at the conflicted locations. The first issue was the core drill cooling water would flow along the embedded utility potentially spreading legacy chemical and radiological contamination to areas of the facility that was unwanted. The second issue was that the REA TS required a substantial structural connection to the B-Cell wall, the cored diameter is larger than the REA TS and these interstitial areas required a full grouted connection. The embedded utilities posed a hazard that if unfilled, the interstitial grout would flow into the embedded utility and destabilize the required grouted structural connection. The B-Cell Cubicles had not been opened in over 10 years. A mockup was completed to install a glovebag over the cubicle opening and then to verify functionality and port requirements. The embedded utility pipe connections in the cubicles were then opened inside a glovebag to verify the correct connection points, connection method for future embedded pipe fill material delivery system, radiological characterization, and air flow to show that the embedded utility pipes were not blocked and would not pressurize. The embedded pipe fill material and grout system was mocked up and run several times to verify methodology and functionality. The B-Cell Cubicles were then reentered and the embedded utility pipe fill was completed using a hand pump. External interferences at the core drill locations were removed using standard interference removal techniques. Internal interferences at the core drill locations were removed using a remote cutting tool that was controlled through the use of the B-Cell crane, remote cameras and remote lighting. The core drilling was mocked up two separate times, the project's purpose built 'Mock Up Facility' was core drilled to verify tolerances and methodology prior to subcontract award, the core drill subcontractor built a 'dummy wall' to train and practice on achieving the demanding tolerances, coordinated alignments between the cores, short term and long term dose shielding techniques, and contamination control techniques. A core drill template, and additional anchoring techniques were used to maximize accuracy and precision of the REA TS core drills. A 'Go/No-Go' gauge was built and installed at each of the four REA TS locations to verify that the core drill met tolerances. The core drill locations were visually inspected using a boroscope. The REA TS installation was mocked up at the projects 'Mock Up Facility' beginning with basic hazard controls, then with intermediate hazard controls, and finally with full occupational safety, industrial hygiene, and radiological controls. The REA TSs were installed and grouted into place. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2017

Investigation of Microbunching Instabilities in Modern Recirculating Accelerators

Particle accelerators are machines to accelerate and store charged particles, such as electrons or protons, to the energy levels for various scientific applications. A collection of charged particles usually forms a particle beam. There are three basic types of particle accelerators: linear accelerators (linac), storage-ring (or circular) accelerators, and recirculating accelerators. In a linac, particles are accelerated and pass through once along a linear or straight beamline. Storage-ring accelerators propel particles around a circular track and repetitively append the energy to the stored beam. The third type, also the most recent one in chronology, the recirculating accelerator, is designed to accelerate the particle beam in a short section of linac, circulate the beam, and then either continue to accelerate for energy boost or decelerate it for energy recovery. The beam properties of a linac machine are set at best by the initial particle sources. For storage rings, the beam equilibria are instead determined by the overall machine design. The modern recirculating machines share with linacs the advantages to both accelerate and preserve the beam with high beam quality, as well as efficiently reuse the accelerating components. The beamline design in such a machine configuration can however be much more complicated than that of linacs. As modern accelerators push toward the high-brightness or high-intensity frontier by demanding particles in a highly charged bunch (about nano-Coulomb per bunch) to concentrate in an ever-decreasing beam phase space (transverse normalized emittance about 1 μm and relative energy spread of the order of 10^-5 in GeV beam energy), the interaction amongst particles via their self-generated electromagnetic fields can potentially lead to coherent instabilities of the beam and thus pose significant challenges to the machine design and operation. In the past decade and a half, microbunching instability (MBI) has been one of the most challenging issues for such high-brightness or high-intensity beam transport, as it would degrade lasing performance in the fourth-generation light sources, reduce cooling efficiency in electron cooling facilities, and compromise the luminosity of colliding beams in lepton or lepton-hadron colliders. The dissertation work will focus on the MBI in modern recirculating electron accelerators. It has been known that the collective interactions, the coherent synchrotron radiation (CSR) and the longitudinal space charge (LSC) forces, can drive MBI. The CSR effect is a collective phenomenon in which the electrons in a curved motion, e.g. a bending dipole, emit radiation at a scale comparable to the micro-bunched structure of the bunch distribution. The LSC effect stems from non-uniformity of the charge distribution, acts as plasma oscillation, and can eventually accumulate an amount of energy modulation when the beam traverses a long section of a beamline. MBI can be seeded by non-uniformity or shot noise of the beam, which originates from granularity of the elementary charge. Through the aforementioned collective effects, the modulation of the bunch sub-structure can be amplified and, once the beam-wave interaction formed a positive feedback, can result in MBI. The problem of MBI has been intensively studied for linac-based facilities and for storage-ring accelerators. However, systematic studies for recirculation machines are still very limited and form a knowledge gap. Because of the much more complicated machine configuration of the recirculating accelerators than that of linacs, the existing MBI analysis needs to be extended to accommodate the high-brightness particle beam transport in modern recirculating accelerators. This dissertation is focused on theoretical investigation of MBI in such machine configuration in the following seven themes: (1) Development and generalization of MBI theory The theoretical formulation has been extended so as to be applicable to a general linear beamline lattice including horizontal and vertical transport bending elements, and beam acceleration or deceleration. These featured generalizations are required for MBI analysis in recirculation accelerators. (2) Construction of CSR impedance models In addition to the steady-state CSR interaction, it has been found that the exit transient effect (or CSR drift) can even result in more serious MBI in high-brightness recirculation arcs. The onedimensional free-space CSR impedances, especially the exit transients, are derived. The steady-state CSR impedance is also extended to non-ultrarelativistic beam energy for MBI analysis of low-energy merger sections in recirculating accelerators. (3) Numerical implementation of the derived semi-analytical formulation This includes the development of a semi-analytical Vlasov solver for MBI analysis, and also benchmarking of the solver against massive particle tracking simulations. (4) Exploration of multistage amplification behavior of CSR microbunching development The CSR-induced MBI acts as an amplifier, which amplifies the sub-bunch modulation of a beam. The amplification is commonly quantified by the amplification gain. A beam transport system can be considered as a cascaded amplifier. Unlike the two-stage amplification of four-dipole bunch compressor chicanes employed in linacs, the recirculation arcs, which are usually constituted by several tens of bending magnets, show a distinguishing feature of up to six-stage microbunching amplification for our example arc lattices. That is, the maximal CSR amplification gain can be proportional to the peak bunch current up to sixth power. A method to compare lattice performance has been developed in terms of gain coefficients, which nearly depend on the lattice properties only. This method has also proven to be an effective way to quantify the current dependence of the maximal (5) Control of CSR MBI in multibend transport or recirculation arcs The existing mitigation schemes of MBI mostly aim to linac-based accelerators and may not be practical to the recirculating accelerator facilities. Thus a set of conditions for suppression of CSR MBI was proposed and examined for example lattices from low (~100 MeV) to high (~1 GeV) energies. (6) Study of more aspects of microbunched structures in beam phase spaces For a cascaded amplifier in circuit electronics, the total amplification gain can be estimated as the product of individual gains. In a beam transport line of an accelerator, the (scalar) gain multiplication was examined and found to under-estimate the overall microbunching amplification. The concept of gain matrix was developed, which includes the density, energy and transverse-longitudinal modulations in a beam phase space, and used to analyze MBI for a proposed recirculating machine. Throughout the gain matrix approach, it reasonably gives the upper limit of spectral MBI gain curves. This extended analysis can be employed to study multi-pass recirculation. (7) Study of MBI for magnetized beams Driven by a recent energy-recovery-linac based cooler design for electron cooling at Jefferson Lab Electron-Ion Collider Project, the generalized theoretical formulation for MBI to a transversely coupled beam has been developed and applied to this study. A magnetized beam in general features non-zero canonical angular momentum, thus considered to be a transversely coupled beam. A novel idea of utilizing magnetized beam transport was proposed for improvement of cooling efficiency and possible mitigation of collective effects. A concern of MBI regarding this design was studied and excluded. The large transverse beam size associated with the beam magnetization is found to help suppress MBI via the transverse-longitudinal correlation.

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