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At least 91 records · Page 5

Phase II Field Demonstration at Lansing Smith Generating Plant, Southport, Florida (Final Report)

The Final Technical Report: Field Test Design and Pressure Management Strategies for Phase II Field Demonstration of Optimal Pressure Control, Plume Management, and Produced Water Strategies presents the culmination of multi-year efforts under the U.S. Department of Energy’s Brine Extraction and Storage Test (BEST) program, led by EPRI in partnership with Gulf Power at the Plant Smith site near Panama City, Florida. The project was designed to evaluate and demonstrate the technical feasibility of managing subsurface pressures and fluid movement associated with large-scale CO₂ injection, using low-salinity water as a proxy fluid. Through a combination of field injection testing, reservoir modeling, and optimization studies, the research team developed and refined active and passive brine extraction strategies aimed at controlling injection-induced pressure buildup, mitigating risks of fault activation, and managing plume migration. The field demonstration incorporated a new injection well (TIW-2), a new monitoring/extraction well (TEMW-A), and an existing well (TIW-1) repurposed for passive pressure relief. Complementary geophysical monitoring was designed to track plume development and evaluate the effectiveness of pressure management strategies. The report details the integrated workflow encompassing wellfield development, regulatory permitting, model calibration, and survey design. It includes a comprehensive electromagnetic (EM) modeling and inversion study used to develop a cost-effective, time-lapse geophysical monitoring plan capable of imaging the evolving low-salinity plume within the highly saline Lower Tuscaloosa formation. Reservoir simulation results guided the design of the 17-month injection program and the timing of active extraction to maintain formation pressures below a hypothetical fault reactivation threshold. Supporting analyses evaluated potential injectivity risks related to clay fines migration and geochemical incompatibility, identifying practical mitigation measures such as salinity and pH control. The final design recommends a series of crosswell EM surveys—conducted before, during, and after injection—as the most effective and economical approach for plume imaging, supplemented by continuous downhole pressure and flow monitoring. Collectively, the study provides a field-ready framework for cost-effective pressure management and monitoring in support of future CO₂ storage projects.

01 COAL, LIGNITE, AND PEAT↗

Grid-Scale, Long-Term Energy Storage: Repurposing Hydrocarbon Reservoirs, Resources, and Infrastructure to Store CO 2 and Heat (Mid-Project Progress Report)

The purpose of this project is to create a viable business case for CO 2 capture, use, and storage (CCUS) by enabling grid-scale, long-term energy storage in mature hydrocarbon reservoirs and saline aquifers. Our CCUS approach, called the Earth Battery, is based on patents issued to Lawrence Livermore National Laboratory on Multi-Fluid Geo-Energy and Earth Battery Systems and to the University of Minnesota and TerraCOH, Inc. on CO 2 Plume Geothermal (CPG). Excess energy can be used to compress and store CO 2 and hot brine underground in permeable formations, including: (1) saline aquifers suitable for geologic CO 2 storage (GCS) and (2) hydrocarbon reservoirs, the focus of this project featuring a pilot test in a mature oil and natural gas (NG) field. Excess energy can come in the form of excess electricity from the grid or excess NG that does not have a market. It can also come in the form of excess heat from other sources, such as from the heat of air or CO 2 compression or from a nearby base-load power plant or solar thermal energy (STE) facility, when heat is not needed to generate electricity from those sources.

15 GEOTHERMAL ENERGY↗

Demonstration of Novel Monitoring Techniques for a North Dakota Carbon Capture and Storage Project

Conference paper presented at 16th International Conference on Greenhouse Gas Control Technologies (GHGT-16), Lyon, France, October 23–27, 2022. We present an overview and initial results of the novel and sustainable monitoring techniques developed by the Energy & Environmental Research Center (EERC) and its commercial partners associated with the Red Trail Energy (RTE) carbon capture and storage (CCS) project. RTE is an investor-owned 64-million-gallon dry mill ethanol production plant in Richardton, North Dakota, USA. Currently in operation as of June 2022, the RTE facility will capture 180,000 tonnes of carbon dioxide (CO 2 ) annually (>99% CO 2 ) that is otherwise emitted during the ethanol production process and inject it into the Broom Creek Formation, a deep saline aquifer, for permanent on-site geologic storage.

01 COAL, LIGNITE, AND PEAT↗

A clustering-based approach to ocean model–data comparison around Antarctica

The Antarctic Continental Shelf seas (ACSS) are a critical, rapidly changing element of the Earth system. Analyses of global-scale general circulation model (GCM) simulations, including those available through the Coupled Model Intercomparison Project, Phase 6 (CMIP6), can help reveal the origins of observed changes and predict the future evolution of the ACSS. However, an evaluation of ACSS hydrography in GCMs is vital: previous CMIP ensembles exhibit substantial mean-state biases (reflecting, for example, misplaced water masses) with a wide inter-model spread. Because the ACSS are also a sparely sampled region, grid-point-based model assessments are of limited value. Our goal is to demonstrate the utility of clustering tools for identifying hydrographic regimes that are common to different source fields (model or data), while allowing for biases in other metrics (e.g., water mass core properties) and shifts in region boundaries. We apply K-means clustering to hydrographic metrics based on the stratification from one GCM (Community Earth System Model version 2; CESM2) and one observation-based product (World Ocean Atlas 2018; WOA), focusing on the Amundsen, Bellingshausen and Ross seas. When applied to WOA temperature and salinity profiles, clustering identifies “primary” and “mixed” regimes that have physically interpretable bases. For example, meltwater-freshened coastal currents in the Amundsen Sea and a region of high-salinity shelf water formation in the southwestern Ross Sea emerge naturally from the algorithm. Both regions also exhibit clearly differentiated inner- and outer-shelf regimes. The same analysis applied to CESM2 demonstrates that, although mean-state model biases in water mass T–S characteristics can be substantial, using a clustering approach highlights that the relative differences between regimes and the locations where each regime dominates are well represented in the model. CESM2 is generally fresher and warmer than WOA and has a limited fresh-water-enriched coastal regimes. Given the sparsity of observations of the ACSS, this technique is a promising tool for the evaluation of a larger model ensemble (e.g., CMIP6) on a circum-Antarctic basis.

54 ENVIRONMENTAL SCIENCES↗

SUBTASK 1.6 – BASIN ELECTRIC CARBON STORAGE RESEARCH PROJECT: NOVEL MONITORING TECHNIQUES

The Energy & Environmental Research Center (EERC) conducted baseline activities associated with an applied research project at Basin Electric Power Cooperative’s (Basin’s) carbon capture and storage (CCS) site in Beulah, North Dakota, to establish novel carbon storage-monitoring techniques as commercial methods under Cooperative Agreement No. DE-FE0024233, Subtask 1.6. The following report summarizes the baseline activities performed and briefly describes the subsequent (operational monitoring) activities that have been proposed to the U.S. Department of Energy (DOE) as part of the overall project to develop and demonstrate novel monitoring techniques at North America’s largest permitted CCS operation. Dakota Gasification Company (DGC), a wholly owned subsidiary of Basin, owns and operates the Great Plains Synfuels Plant (GPSP) approximately 5 miles northwest of the town of Beulah, North Dakota (Figure 1). In 2023, DGC received approval from the North Dakota Industrial Commission (NDIC) to develop a storage facility on-site for injecting a stream of carbon dioxide (CO2) captured from GPSP. DGC will transport the captured CO2 stream with approximately 6.8 miles of transmission lines that extend north of GPSP and inject >1 million tonnes (MMt) of CO2 annually (>1 MMt/yr) over a 12-year period with up to six underground injection control (UIC) Class VI-compliant injection wells completed in the Broom Creek Formation, a predominantly sandstone reservoir and saline aquifer underlying GPSP. The Broom Creek Formation lies approximately 5900 feet (ft) below ground surface (bgs) at GPSP. The commercial scale (i.e., >1 MMt/yr) of DGC’s permitted carbon storage project is ideal for developing and testing the novel monitoring techniques included within Subtask 1.6. The goals of this project are to demonstrate 1) the cost-effectiveness of novel monitoring technologies included as part of this research, 2) technology capability for tracking the CO2 plume and/or associated pressure response in the subsurface and monitoring out-of-zone migration, and 3) compliance with UIC Class VI program requirements. The research activities proposed for the overall project include 1) design of an automated, integrated, modular (AIM) monitoring station; 2) time-lapse electromagnetic (EM) field surveys; 3) drone-based surveillance studies; 4) time-lapse monitoring with seismic methods; 5) advanced wellbore-monitoring methods; 6) deployment of an AIM monitoring network; 7) EM monitoring of CO2 with real-time data processing; 8) continued seasonal drone-based surveillance studies; 9) seismic monitoring with passive and active surveys; and 10) wellbore monitoring with nuclear magnetic resonance (NMR) for near-surface characterization. Completion of Activities 1.0–5.0 (baseline activities) are described in this report. Upon authorization of funding by DOE, the EERC will initiate Activities 6.0– 10.0 (operational monitoring activities). Current state-of-the-art (SOA) carbon storage-monitoring techniques require countless labor hours dedicated to the acquisition of data. Once data are gathered, these SOA techniques often rely on commercial facilities to process raw data from the field. However, it is anticipated that next-generation monitoring techniques, such as those being demonstrated, will lower acquisition footprints, be less operationally intensive, and improve data acquisition efficiencies. These new techniques are more conducive to the application of machine learning, artificial intelligence, and automation, thus providing a pathway for integration into active control systems, informing site operability, and improving the integration of data for future CCS projects across the United States. Additionally, reclaimed and active mining lands are present within the project site, creating a unique opportunity to demonstrate the effectiveness of remote sensing and surface-based geophysics monitoring techniques at similar project sites that may include disturbed, unconsolidated, or actively excavated near-surface environments. The efforts included in the overall project will produce necessary designs, learnings, and data acquired during the baseline and operational monitoring periods that are necessary for time-lapse demonstration and validation of the described monitoring techniques. In addition, it is anticipated that the monitoring technologies included in this study will be compliant with UIC Class VI requirements to enable the potential for implementation at other CCS sites across the United States.

42 ENGINEERING↗

Salinity tolerance mechanisms of an Arctic Pelagophyte using comparative transcriptomic and gene expression analysis

Abstract Little is known at the transcriptional level about microbial eukaryotic adaptations to short-term salinity change. Arctic microalgae are exposed to low salinity due to sea-ice melt and higher salinity with brine channel formation during freeze-up. Here, we investigate the transcriptional response of an ice-associated microalgae over salinities from 45 to 8. Our results show a bracketed response of differential gene expression when the cultures were exposed to progressively decreasing salinity. Key genes associated with salinity changes were involved in specific metabolic pathways, transcription factors and regulators, protein kinases, carbohydrate active enzymes, and inorganic ion transporters. The pelagophyte seemed to use a strategy involving overexpression of Na + -H + antiporters and Na + -Pi symporters as salinity decreases, but the K + channel complex at higher salinities. Specific adaptation to cold saline arctic conditions was seen with differential expression of several antifreeze proteins, an ice-binding protein and an acyl-esterase involved in cold adaptation.

59 BASIC BIOLOGICAL SCIENCES↗

Partial resolution of sources of n-alkanes in the saline portion of the Parachute Creek Member, Green River Formation (Piceance Creek Basin, Colorado)

Systematic variations in the 13C contents of individual extractable n-alkanes (C16-C29) can be modelled quantitatively and interpreted as indicating contributions from at least five distinct sources. These appear to be cyanobacterial (C16-C18, delta 13C = -37% vs PDB), phytoplanktonic (C16-C23, delta = -32%), chemoautotrophic bacterial (C20-C29, delta = -38%), phytoplanktonic or heterotrophic bacterial (C20-C29, delta = -30%), and vascular plants (C23-C29, delta = -29%). Hydrous pyrolysis of related kerogens yields large quantities of additional n-alkanes with different and much more uniform delta values. The latter materials are apparently derived from the thermolysis of aliphatic biopolymers whose presence in the Green River Oil Shale has been recognized visually.

Non-NASA Center↗

Evidence of thyroxine formation following iodine administration in Sprague-Dawley rats

Iodine (I2) has been proposed to be used as a water disinfectant on the manned space station. Previous work has shown that subchronic administration of I2 to Sprague-Dawley rats in drinking water significantly increases plasma thyroxine/triiodothyronine (T4/T3) levels. This is not observed with iodide (I-) treatment. The present study addresses the possibility that I2 reacts with deiodinated T4 metabolites in the gastrointestinal tract to resynthesize T4. Incubation of diiodothyronine (T2), T3, or reverse T3 with I2 in phosphate-buffered saline resulted in the formation of T4 as measured by radioimmunoassay. Washes from the initial segments of the small intestine of the rat show that substrates are present that react with I2 to produce T4. Single oral doses of I2 to rats produced significant dose-related increases in serum T4 and decreases in T3 concentrations after 2 h. Administration of an equivalent dose of I- did not alter significantly plasma T4 concentrations. Higher concentrations of a radioactive substance that bound a T4-specific antibody are present in plasma of animals treated with 125I2 compared to 125I-. These data support the hypothesis that I2 reacts with metabolites of thyroid hormone in the gastrointestinal tract to resynthesize T4 and elevate its levels in blood.

NASA Discipline Environmental Health↗

Observations and theoretical studies of microwave emission from thin saline ice

Time-dependent changes in microwave emissions from thin artificial and natural saline ices are reported. There is a sharp rise in surface temperature when ice is between 1 and 2 cm thick, apparently unrelated to any environmental changes. The brightness temperature Tb decreases at 37 GHz during or just after the surface temperature rise, and there is an initial increase in Tb with increasing ice thickness followed by substantial decreases at the higher studied frequencies. The maximum Tb values were higher than those previously reported for young ice. Tb was also found to be much more sensitive to variations in ice properties at horizontal polarization than at vertical polarization. The most likely explanation for the observed rise in surface temperature and decrease in Tb was the formation of a salinity-enhanced ice of brine surface layer caused by the upwards transport of brine as the ice grows.

Wensnahan, Mark R.↗

NRAP Task 5: Preliminary Evaluation of the Cost of Responding to a Hypothetical Leakage Scenario Using the NRAP/SMART TALES Model and other NRAP Tools

Poster presentation illustrating the use of tools developed as part of Task 5 of Phase 3 of NRAP to estimate the technical performance and costs of implementing remedial responses to address a leak of fluid out of the storage formation at a CO2 saline storage project. Presented at the 2024 FECM - NETL Carbon Management Research Project Review Meeting, 5-9 August, 2024, Pittsburgh, PA.

Morgan, David↗

Variability and Anomalous Trends in the Global Sea Ice Cover

The advent of satellite data came fortuitously at a time when the global sea ice cover has been changing rapidly and new techniques are needed to accurately assess the true state and characteristics of the global sea ice cover. The extent of the sea ice in the Northern Hemisphere has been declining by about -4% per decade for the period 1979 to 2011 but for the period from 1996 to 2010, the rate of decline became even more negative at -8% per decade, indicating an acceleration in the decline. More intriguing is the drastically declining perennial sea ice area, which is the ice that survives the summer melt and observed to be retreating at the rate of -14% per decade during the 1979 to 2012 period. Although a slight recovery occurred in the last three years from an abrupt decline in 2007, the perennial ice extent was almost as low as in 2007 in 2011. The multiyear ice, which is the thick component of the perennial ice and regarded as the mainstay of the Arctic sea ice cover is declining at an even higher rate of -19% per decade. The more rapid decline of the extent of this thicker ice type means that the volume of the ice is also declining making the survival of the Arctic ice in summer highly questionable. The slight recovery in 2008, 2009 and 2010 for the perennial ice in summer was likely associated with an apparent cycle in the time series with a period of about 8 years. Results of analysis of concurrent MODIS and AMSR-E data in summer also provide some evidence of more extensive summer melt and meltponding in 2007 and 2011 than in other years. Meanwhile, the Antarctic sea ice cover, as observed by the same set of satellite data, is showing an unexpected and counter intuitive increase of about 1 % per decade over the same period. Although a strong decline in ice extent is apparent in the Bellingshausen/ Amundsen Seas region, such decline is more than compensated by increases in the extent of the sea ice cover in the Ross Sea region. The results of analysis of MODIS, AMSR-E and SSM/I data reveal that the sea ice production rate at the coastal polynyas along the Ross Ice Shelf has been increasing since 1992. This also means that the salinization rate and the formation of bottom water in the region are going up as well. Simulation studies indicate that the stronger production rate is likely associated with the ozone hole that has caused a deepening of the lows in the West Antarctic region and therefore stronger winds off the Ross Ice Shelf. Stronger winds causes larger coastal polynyas near the shelf and hence an enhanced ice production in the region during the autumn and winter period. Results of analysis of temperature data from MODIS and AMSR-E shows that the area and concentration of the sea ice cover are highly correlated with surface temperature for both the Arctic and Antarctic, especially in the seasonal regions where the correlation coefficients are about 0.9. Abnormally high sea surface temperatures (SSTs) and surface ice temperatures (SITs) were also observed in 2007 and 2011when drastic reductions in the summer ice cover occurred, This phenomenon is consistent with the expected warming of the upper layer of the Arctic Ocean on account of ice-albedo feedback. Changes in atmospheric circulation are also expected to have a strong influence on the sea ice cover but the results of direct correlation analyses of the sea ice cover with the Northern and the Southern Annular Mode indices show relatively weak correlations, This might be due in part to the complexity of the dynamics of the system that can be further altered by some phenomena like the Antarctic Circumpolar Wave and extra polar processes like the El Nino Southern Oscillation (ENSO) and the Pacific Decadal Oscillation (POD),

Comiso, Josefino C.↗

Surface Reactivity Analysis of the Crude Oil–Brine–Limestone Interface for a Comprehensive Understanding of the Low-Salinity Waterflooding Mechanism

Low-salinity waterflooding (LSWF) has proven to improve oil recovery in carbonate formations through rock wettability alteration, although the underlying mechanism remains elusive. Multivalent ionic exchange and calcite dissolution have usually been investigated using geochemical analysis in secondary coreflooding. In this work, coreflooding, in tertiary mode, coupled with a surface reactivity analysis approach was employed to investigate the interplay of wettability alteration mechanisms such as mineral dissolution, electrostatic bond attraction, and the effect of pH at in situ conditions. Improved oil recovery (IOR) in tertiary mode observed by coreflooding in Indiana limestone rocks showed an ionic strength dependence, that is, reducing brine ionic strength resulted in an increase in oil recovery. Coreflooding results showed that the seawater and low-salinity brines deprived of Mg 2+ ions resulted in the lowest IOR in tertiary mode, indicating the significance of Mg 2+ on IOR in limestone rocks. Similar results were observed through the contact angle measurement showing the limestone rock wettability state dependence on ionic strength and the effect of Mg2+ ions. Surface reactivity analysis showed an increase in solution pH, Ca 2+ and Mg 2+ ions concentration in the effluent solution from the coreflooding in tertiary mode using low salinity brines (about 40 and 20% increase in the effluent composition for Ca 2+ and Mg 2+ , respectively). These changes in solution composition were used to calculate the in situ oil–brine and rock–brine zeta potential using a validated surface complexation model, showing the changes of zeta potential as brine is injected into limestone rocks. The results show that using seawater-like brine in tertiary mode resulted in no mineral dissolution or ionic exchange. However, improved oil recovery (IOR) using such seawater-like brine was due to wettability alteration caused by reduced electrostatic bond attraction associated with Mg 2+ ions [from 2.6 × 10 –13 (mol/m 2 ) 2 for formation water salinity to 1.5 × 10 –13 (mol/m 2 ) 2 for seawater salinity]. Using low-salinity brines in tertiary mode improved oil recovery by mineral dissolution, resulting in oil desorption and an increase in solution pH. Finally, the increase in solution pH also resulted in reduced electrostatic bond attraction which lead to rock wettability alteration using low-salinity brines.

04 OIL SHALES AND TAR SANDS↗

Effects of Carbonic Acid-Rock Interactions on CO2/Brine Multiphase Flow Properties in the Upper Minnelusa Sandstones

Summary Carbon dioxide (CO2) injection into a deep saline aquifer can dissolve into formation brine and generate carbonic acid. The resulting acid can drive fluid-rock geochemical reactions. The impact of these fluid-rock geochemical reactions on porosity, permeability, and multiphase flow responses is relevant to the determination of CO2 storage capacity of deep saline aquifers. In this research, carbonic acid flooding experiments were performed on core samples consisting of poorly sorted, quartz-rich sand with laminated bedding from a possible CO2 storage target in northwest Wyoming. Complementary pre- and post-injection porosity and permeability, thin-section, Brunauer-Emmett-Teller (BET) surface area, mercury intrusion capillary pressure (MICP), and time-domain nuclear magnetic resonance (TD-NMR) measurements were conducted. Overall, both core porosity and permeability increased after a 7-day carbonic acid injection, from 6.2 to 8.4% and 1.6 to 3.7 md, respectively. We attributed these changes to carbonate mineral dissolution, which was evidenced by the effluent brine geochemistry, pore-throat size distribution (PTSD), and BET surface area. To be more specific, within the more-permeable section of core samples containing larger pore size, the permeability increment is apparent due to dolomite mineral grains and cements dissolution. However, for the lower-permeability section corresponding to the smaller pore size, mineral precipitation possibly lessened dissolution effects, leading to insignificant petrophysical properties changes. Consequently, the observed heterogeneous carbonic acid-rock interactions resulted in alterations of CO2/brine relative permeability (i.e., the initial CO2 saturation decreased and the CO2 flow capacity was enhanced). This research provides a fundamental understanding regarding effects of fluid-rock reactions on changes in static and multiphase flow properties of eolian sandstones, which lays the foundation for more accurate prediction/simulation of CO2 injection into deep saline aquifers.

Engineering↗

Maximizing the Proppant Carrying and Viscoelastic Properties of the Bakken Hypersaline-Produced Water with High-Viscosity Friction Reducers for Sustainable Applications

Summary The development and production of unconventional reservoirs, such as the Bakken Formation, have become a resolved mystery for operators in North America since the arrival and advancement of horizontal drilling and hydraulic fracturing technologies. As a result, unconventional reservoir assets became the central focus of the oil and gas industry at the state, national, and global levels. The produced water from these activities in the Bakken Formation have high salt contents (110,000–350,000 ppm) total dissolve solids (TDS) and can pose significant challenges to the environment if not treated. Deep injection into disposal wells is the routine method used to get rid of the Bakken produced water. However, there have been some concerns that unrestrained injections, in addition to polluting the groundwater, could potentially lead to seismic activities either at the time of injection or in the near future. To diminish the environmental impacts that may be associated with induced seismicity, including the reduction of the costs of water acquisition, the produced water can be treated and reused in the hydraulic fracturing processes. Also, the treated water could be used for irrigation purposes, for power generation, and coal mining operations. The issues of waste water and residual oil high in TDS are challenges yet to be effectively addressed despite preceding research and studies on advancing produced water technologies. The goal of this study is to explore all applicable ways by which the produced water from the Bakken Formation can serve as a replacement base fluid for use with polymers like the high-viscosity friction reducers (HVFRs) to create hydraulic fracturing fluids that can be stable with reservoir conditions and also be able to minimize environmental impacts and cost of operations. Experimental investigations using the high-salinity produced water from the Bakken Formation with HVFRs were carried out. The studies included a base case that served as a bench mark for comparing the effectiveness of the other scenarios. The results indicate that the Bakken hyper-saline produced water can withstand effect of heavy metals, salinity, hardness and remain stable through different shear rates (66–330 s−1) when treated with higher dosages [4–8 gal/1,000 gal (gpt)] of HVFRs. Filtration and dilution were the only methods used on the Bakken Formation produced wate for this research

Engineering↗

Coupled ice-ocean variability in the Greenland Sea

A major surface feature of the Greenland Sea during winter is the frequent eastward extension of sea ice south of 75 deg N and an associated embayment to the north. These features are nominally connected with the East Greenland Current, and both the promontory) and the embayment are readily apparent on climatic ice charts. However, there are significant changes in these features on time-scales as short as a few days. Using a combination of satellite microwave images (SSM/I) of ice cover, meteorological data and in situ velocity, temperature and salinity records, we relate the ice distribution and its changes to the developing structure and circulation of the upper ocean during winter 1988-1989. Our measurements illustrate the preconditioning that leads to convective overturn, which in turn brings warmer water to the surface and results in the rapid disappearance of ice. In particular, the surface was cooled to the freezing point by early December and the salinity then increased through ice formation (about 0.016 m/d) and brine rejection. Once the vertical density gradient was sufficiently eroded, a period of high heat flux (greater than 300 W/sq m) in late January provided enough buoyancy loss to convectively mix the upper water column to at least 200 m. We estimate vertical velocities at about 3 cm/s downward during the initial sinking. The deepening of the thermocline raised surface temperatures by over 1 C resulting in nearly 1.5 x 105 km of ice-melt within two days. Average rates of ice retreat are about 2 km/d southwestward, generally consistent with a wind-driven flow. Comparison of hydrographic surveys from before and after the overturning indicate the fresh water was advected out of the area, possibly to the south and east of our moorings.

Roach, A. T.↗

Microbial colonization and persistence in deep fractured shales is guided by metabolic exchanges and viral predation

Background: Microbial colonization of subsurface shales following hydraulic fracturing offers the opportunity to study coupled biotic and abiotic factors that impact microbial persistence in engineered deep subsurface ecosystems. Shale formations underly much of the continental USA and display geographically distinct gradients in temperature and salinity. Complementing studies performed in eastern USA shales that contain brine-like fluids, here we coupled metagenomic and metabolomic approaches to develop the first genome-level insights into ecosystem colonization and microbial community interactions in a lower-salinity, but high-temperature western USA shale formation. Results: We collected materials used during the hydraulic fracturing process (i.e., chemicals, drill muds) paired with temporal sampling of water produced from three different hydraulically fractured wells in the STACK (Sooner Trend Anadarko Basin, Canadian and Kingfisher) shale play in OK, USA. Relative to other shale formations, our metagenomic and metabolomic analyses revealed an expanded taxonomic and metabolic diversity of microorganisms that colonize and persist in fractured shales. Importantly, temporal sampling across all three hydraulic fracturing wells traced the degradation of complex polymers from the hydraulic fracturing process to the production and consumption of organic acids that support sulfate- and thiosulfate-reducing bacteria. Furthermore, we identified 5587 viral genomes and linked many of these to the dominant, colonizing microorganisms, demonstrating the key role that viral predation plays in community dynamics within this closed, engineered system. Lastly, top-side audit sampling of different source materials enabled genome-resolved source tracking, revealing the likely sources of many key colonizing and persisting taxa in these ecosystems. Conclusions: These findings highlight the importance of resource utilization and resistance to viral predation as key traits that enable specific microbial taxa to persist across fractured shale ecosystems. We also demonstrate the importance of materials used in the hydraulic fracturing process as both a source of persisting shale microorganisms and organic substrates that likely aid in sustaining the microbial community. Moreover, we showed that different physicochemical conditions (i.e., salinity, temperature) can influence the composition and functional potential of persisting microbial communities in shale ecosystems. Together, these results expand our knowledge of microbial life in deep subsurface shales and have important ramifications for management and treatment of microbial biomass in hydraulically fractured wells.

59 BASIC BIOLOGICAL SCIENCES↗

Functional Insights of Salinity Stress-Related Pathways in Metagenome-Resolved Methanothrix Genomes

Recently, methanogenic archaea belonging to the genus Methanothrix were reported to have a fundamental role in maintaining stable ecosystem functioning in anaerobic bioreactors under different configurations/conditions. In this study, we reconstructed three Methanothrix metagenome-assembled genomes (MAGs) from granular sludge collected from saline upflow anaerobic sludge blanket (UASB) reactors, where Methanothrix harundinacea was previously implicated with the formation of compact and stable granules under elevated salinity levels (up to 20 g/L Na + ). Genome annotation and pathway analysis of the Methanothrix MAGs revealed a genetic repertoire supporting their growth under high salinity. Specifically, the most dominant Methanothrix (MAG_279), classified as a subspecies of Methanothrix_A harundinacea_D, had the potential to augment its salinity resistance through the production of different glycoconjugates via the N-glycosylation process, and via the production of compatible solutes as N ε -acetyl-β-lysine and ectoine. The stabilization and reinforcement of the cell membrane via the production of isoprenoids was identified as an additional stress-related pathway in this microorganism. The improved understanding of the salinity stress-related mechanisms of M. harundinacea highlights its ecological niche in extreme conditions, opening new perspectives for high-efficiency methanisation of organic waste at high salinities, as well as the possible persistence of this methanogen in highly-saline natural anaerobic environments.

59 BASIC BIOLOGICAL SCIENCES↗