Browsing by Author "Mitchell, Andrew C."
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Item Bacterially induced calcium carbonate precipitation and strontium coprecipitation in a porous media flow system(2013-02) Lauchnor, Ellen G.; Schultz, Logan N.; Bugni, S.; Mitchell, Andrew C.; Cunningham, Alfred B.; Gerlach, RobinStrontium-90 is a principal radionuclide contaminant in the subsurface at several Department of Energy sites in the Western U.S., causing a threat to groundwater quality in areas such as Hanford, WA. In this work, we used laboratory-scale porous media flow cells to examine a potential remediation strategy employing coprecipitation of strontium in carbonate minerals. CaCO3 precipitation and strontium coprecipitation were induced via ureolysis by Sporosarcina pasteurii in two-dimensional porous media reactors. An injection strategy using pulsed injection of calcium mineralization medium was tested against a continuous injection strategy. The pulsed injection strategy involved periods of lowered calcite saturation index combined with short high fluid velocity flow periods of calcium mineralization medium followed by stagnation (no-flow) periods to promote homogeneous CaCO3 precipitation. By alternating the addition of mineralization and growth media the pulsed strategy promoted CaCO3 precipitation while sustaining the ureolytic culture over time. Both injection strategies achieved ureolysis with subsequent CaCO3 precipitation and strontium coprecipitation. The pulsed injection strategy precipitated 71−85% of calcium and 59% of strontium, while the continuous injection was less efficient and precipitated 61% of calcium and 56% of strontium. Over the 60-day operation of the pulsed reactors, ureolysis was continually observed, suggesting that the balance between growth and precipitation phases allowed for continued cell viability. Our results support the pulsed injection strategy as a viable option for ureolysis-induced strontium coprecipitation because it may reduce the likelihood of injection well accumulation caused by localized mineral plugging while Sr coprecipitation efficiency is maintained in field-scale applications.Item Biofilm enhanced subsurface sequestration of supercritical CO2(2009-01) Mitchell, Andrew C.; Phillips, Adrienne J.; Hiebert, Dwight Randall; Gerlach, Robin; Cunningham, Alfred B.In order to develop subsurface CO2 storage as a viable engineered mechanism to reduce the emission of CO2 into the atmosphere, any potential leakage of injected supercritical CO2 (SC-CO2) from the deep subsurface to the atmosphere must be reduced. Here, we investigate the utility of biofilms, which are microorganism assemblages firmly attached to a surface, as a means of reducing the permeability of deep subsurface porous geological matrices under high pressure and in the presence of SC-CO2, using a unique high pressure (8.9 MPa), moderate temperature (32 °C) flow reactor containing 40 millidarcy Berea sandstone cores. The flow reactor containing the sandstone core was inoculated with the biofilm forming organism Shewanella fridgidimarina. Electron microscopy of the rock core revealed substantial biofilm growth and accumulation under high-pressure conditions in the rock pore space which caused >95% reduction in core permeability. Permeability increased only slightly in response to SC-CO2 challenges of up to 71 h and starvation for up to 363 h in length. Viable population assays of microorganisms in the effluent indicated survival of the cells following SC-CO2 challenges and starvation, although S. fridgidimarina was succeeded by Bacillus mojavensis and Citrobacter sp. which were native in the core. These observations suggest that engineered biofilm barriers may be used to enhance the geologic sequestration of atmospheric CO2.Item Biogeochemical Connectivity Between Freshwater Ecosystems beneath the West Antarctic Ice Sheet and the Sub‐Ice Marine Environment(2020-03) Vick‐Majors, Trista J.; Michaud, Alexander B.; Skidmore, Mark L.; Turetta, Clara; Barbante, Carlo; Christner, Brent C.; Dore, John E.; Christianson, Knut; Mitchell, Andrew C.; Achberger, Amanda M.; Mikucki, Jill A.; Priscu, John C.Although subglacial aquatic environments are widespread beneath the Antarctic ice sheet, subglacial biogeochemistry is not well understood, and the contribution of subglacial water to coastal ocean carbon and nutrient cycling remains poorly constrained. The Whillans Subglacial Lake (SLW) ecosystem is upstream from West Antarctica's Gould‐Siple Coast ~800 m beneath the surface of the Whillans Ice Stream. SLW hosts an active microbial ecosystem and is part of an active hydrological system that drains into the marine cavity beneath the adjacent Ross Ice Shelf. Here we examine sources and sinks for organic matter in the lake and estimate the freshwater carbon and nutrient delivery from discharges into the coastal embayment. Fluorescence‐based characterization of dissolved organic matter revealed microbially driven differences between sediment pore waters and lake water, with an increasing contribution from relict humic‐like dissolved organic matter with sediment depth. Mass balance calculations indicated that the pool of dissolved organic carbon in the SLW water column could be produced in 4.8 to 11.9 yr, which is a time frame similar to that of the lakes’ fill‐drain cycle. Based on these estimates, subglacial lake water discharged at the Siple Coast could supply an average of 5,400% more than the heterotrophic carbon demand within Siple Coast embayments (6.5% for the entire Ross Ice Shelf cavity). Our results suggest that subglacial discharge represents a heretofore unappreciated source of microbially processed dissolved organic carbon and other nutrients to the Southern Ocean.Item Biogeochemistry and microbial diversity in the marine cavity beneath the McMurdo Ice Shelf, Antarctica(2016-03) Vick-Majors, Trista J.; Achberger, Amanda M.; Santibanez, Pamela A.; Dore, John E.; Hodson, Timothy; Michaud, Alexander B.; Christner, Brent C.; Mikucki, Jill A.; Skidmore, Mark L.; Powell, Ross; Adkins, W. Peyton; Barbante, Carlo; Mitchell, Andrew C.; Scherer, Reed; Priscu, John C.Ice shelves surround ∼ 75% of Antarctica's coastline and are highly sensitive to climate change; several have recently collapsed and others are predicted to in the near future. Marine waters beneath ice shelves harbor active ecosystems, while adjacent seas can be important areas of bottom water formation. Despite their oceanographic significance, logistical constraints have resulted in few opportunities to directly sample sub-ice shelf cavities. Here, we present the first data on microbial diversity and biogeochemistry beneath the McMurdo Ice Shelf (MIS) near Ross Island, Antarctica. Physicochemical profiles obtained via a 56 m deep borehole through the MIS revealed three vertically layered water masses (Antarctic Surface Water [AASW], Ice Shelf Water [ISW], and modified High Salinity Shelf Water [mHSSW]). Metabolically active, moderately diverse (Shannon diversity from 2.06 to 5.74) microbial communities were detected in the AASW and mHSSW. Heterotrophic bacterial production and dissolved organic matter concentrations were higher (12–37% and 24%, respectively) in mHSSW relative to AASW. Chemoautotrophic production was 5.3 nmol C L−1 d−1 and 6.0 nmol C L−1 d−1 in the AASW and mHSSW, respectively. Phytoplankton cells were more abundant and larger in the mHSSW sample relative to the AASW, which indicates sinking of phytoplankton produced in surface waters and, together with southerly flowing currents (0.09–0.16 m s−1), horizontal advection of phytoplankton from McMurdo Sound. Advected phytoplankton carbon together with in situ chemoautotrophic production provide important sources of organic matter and other reduced compounds to support ecosystem processes in the dark waters in the ice shelf cavity.Item Design of a meso-scale high pressure vessel for the laboratory examination of biogeochemical subsurface processes(2015-02) Phillips, Adrienne J.; Eldring, Joseph; Hiebert, Dwight Randall; Lauchnor, Ellen G.; Mitchell, Andrew C.; Cunningham, Alfred B.; Spangler, Lee H.; Gerlach, RobinBiocides are critical components of hydraulic fracturing (“fracking†) fluids used for unconventional shale gas development. Bacteria may cause bioclogging and inhibit gas extraction, produce toxic hydrogen sulfide, and induce corrosion leading to downhole equipment failure. The use of biocides such as glutaraldehyde and quaternary ammonium compounds has spurred a public concern and debate among regulators regarding the impact of inadvertent releases into the environment on ecosystem and human health. This work provides a critical review of the potential fate and toxicity of biocides used in hydraulic fracturing operations. We identified the following physicochemical and toxicological aspects as well as knowledge gaps that should be considered when selecting biocides: (1) uncharged species will dominate in the aqueous phase and be subject to degradation and transport whereas charged species will sorb to soils and be less bioavailable; (2) many biocides are short-lived or degradable through abiotic and biotic processes, but some may transform into more toxic or persistent compounds; (3) understanding of biocides’ fate under downhole conditions (high pressure, temperature, and salt and organic matter concentrations) is limited; (4) several biocidal alternatives exist, but high cost, high energy demands, and/or formation of disinfection byproducts limits their use. This review may serve as a guide for environmental risk assessment and identification of microbial control strategies to help develop a sustainable path for managing hydraulic fracturing fluids.Item Diversity, abundance, and potential activity of nitrifying and nitrate-reducing microbial assemblages in a subglacial ecosystem(2011-05) Boyd, Eric S.; Lange, Rachel K.; Mitchell, Andrew C.; Havig, Jeff R.; Lafreniere, M. J.; Shock, Everett L.; Peters, John W.; Skidmore, Mark L.Subglacial sediments sampled from beneath Robertson Glacier (RG), Alberta, Canada, were shown to harbor diverse assemblages of potential nitrifiers, nitrate reducers, and diazotrophs, as assessed by amoA, narG, and nifH gene biomarker diversity. Although archaeal amoA genes were detected, they were less abundant and less diverse than bacterial amoA, suggesting that bacteria are the predominant nitrifiers in RG sediments. Maximum nitrification and nitrate reduction rates in microcosms incubated at 4°C were 280 and 18.5 nmol of N per g of dry weight sediment per day, respectively, indicating the potential for these processes to occur in situ. Geochemical analyses of subglacial sediment pore waters and bulk subglacial meltwaters revealed low concentrations of inorganic and organic nitrogen compounds. These data, when coupled with a C/N atomic ratio of dissolved organic matter in subglacial pore waters of ∼210, indicate that the sediment communities are N limited. This may reflect the combined biological activities of organic N mineralization, nitrification, and nitrate reduction. Despite evidence of N limitation and the detection of nifH, we were unable to detect biological nitrogen fixation activity in subglacial sediments. Collectively, the results presented here suggest a role for nitrification and nitrate reduction in sustaining microbial life in subglacial environments. Considering that ice currently covers 11% of the terrestrial landmass and has covered significantly greater portions of Earth at times in the past, the demonstration of nitrification and nitrate reduction in subglacial environments furthers our understanding of the potential for these environments to contribute to global biogeochemical cycles on glacial-interglacial timescales.Item Engineered applications of ureolytic biomineralization: A review(2013-07) Phillips, Adrienne J.; Gerlach, Robin; Lauchnor, Ellen G.; Mitchell, Andrew C.; Cunningham, Alfred B.; Spangler, Lee H.Microbially induced calcium carbonate (CaCO3) precipitation (MICP) is a widely explored and promising technology for use in various engineering applications. In this review, CaCO3 precipitation induced via urea hydrolysis (ureolysis) is examined for improving construction materials, cementing porous media, hydraulic control, and remediating environmental concerns. The control of MICP is explored through the manipulation of three factors: (1) the ureolytic activity (of microorganisms), (2) the reaction and transport rates of substrates, and (3) the saturation conditions of carbonate minerals. Many combinations of these factors have been researched to spatially and temporally control precipitation. This review discusses how optimization of MICP is attempted for different engineering applications in an effort to highlight the key research and development questions necessary to move MICP technologies toward commercial scale applications.Item Imaging biologically induced mineralization in fully hydrated flow systems(2011) Schultz, Logan N.; Pitts, Betsey; Mitchell, Andrew C.; Cunningham, Alfred B.; Gerlach, RobinA number of proposed technologies involve the controlled implementation of biologically induced carbonate mineral precipitation in the geologic subsurface. Examples include the enhancement of soil stability [1], immobilization of groundwater contaminants such as strontium and uranium [2], and the enhancement of oil recovery and geologic carbon sequestration via controlled permeability reduction [3]. The most significant challenge in these technologies remains to identify and better understand an industrially, environmentally, and economically viable carbonate precipitation route.One of the most promising routes is ureolytic biomineralization, because of the ample availability of urea and the controllable reaction rate. In this process, ureolytic bacteria hydrolyze urea, leading to an increase in pH. In the presence of calcium, this process favors the formation of solid calcium carbonate, as illustrated in the following equations:CO(NH2)2 + H2O → NH2COOH + NH3→ 2 NH3 + CO2 (Urea hydrolysis) (1)2 NH3 + 2 H2O ↔ 2NH4+ + 2OH– (pH increase) (2)CO2 + 2 OH– ↔ CO32– + H2O(Carbonate ion formation) (3)CO32– + Ca2+ ↔ CaCO3 (solid)(Precipitation is favored at high pH) (4)This process relies on molecular-level chemical and biological processes that must be better understood for large-scale implementation.Researchers at the Center for Biofilm Engineering at Montana State University (USA) and Aberystwyth University (UK) have conducted several biomineralization experiments in simulated porous media reactors. Microscopy has proven to be one of the most useful analytical tools in these studies, providing the ability to non-invasively visualize, differentiate, and quantify the various components, including the cells, cell matrix, and mineral precipitates. Because of the possibility of real-time observation and the lack of dehydration artifacts, microscopy has been tremendously useful for elucidating the temporal and spatial relationships of these components.Item Influence of bedrock mineral composition on microbial diversity in a subglacial environment(2013-06) Mitchell, Andrew C.; Lafrenière, M. J.; Skidmore, Mark L.; Boyd, Eric S.Microorganisms in subglacial environments drive the chemical weathering of bedrock; however, the influence of bedrock mineralogy on the composition and activity of microbial assemblages in such environments is poorly understood. Here, using a combination of in situ mineral incubation and DNA fingerprinting techniques, we demonstrate that pyrite is the dominant mineralogical control on subglacial bacterial community structure and composition. In addition, we show that the abundance of Fe in the incubated minerals influences the development of mineral-associated biomass. The ubiquitous nature of pyrite in many common bedrock types and high SO42– concentrations in most glacial meltwaters suggest that pyrite may be a dominant lithogenic control on microbial communities in many subglacial systems. Mineral-based energy may therefore serve a fundamental role in sustaining subglacial microbial populations and enabling their persistence over glacial-interglacial time scales.Item Kinetics of Calcite Precipitation by Ureolytic Bacteria under Aerobic and Anaerobic Conditions(2019-05) Mitchell, Andrew C.; Espinosa-Ortiz, Erika J.; Parks, Stacy L.; Phillips, Adrienne J.; Cunningham, Alfred B.; Gerlach, RobinThe kinetics of urea hydrolysis (ureolysis) and induced calcium carbonate (CaCO3) precipitation for engineering use in the subsurface was investigated under aerobic conditions using Sporosarcina pasteurii (ATCC strain 11859) as well as Bacillus sphaericus strains 21776 and 21787. All bacterial strains showed ureolytic activity inducing CaCO3 precipitation aerobically. Rate constants not normalized to biomass demonstrated slightly higher-rate coefficients for both ureolysis (kurea) and CaCO3 precipitation (kprecip) for B. sphaericus 21776 (kurea=0.10±0.03 h−1, kprecip=0.60±0.34 h−1) compared to S. pasteurii (kurea=0.07±0.02 h−1, kprecip=0.25±0.02 h−1), though these differences were not statistically significantly different. B. sphaericus 21787 showed little ureolytic activity but was still capable of inducing some CaCO3 precipitation. Cell growth appeared to be inhibited during the period of CaCO3 precipitation. Transmission electron microscopy (TEM) images suggest this is due to the encasement of cells and was reflected in lower kurea values observed in the presence of dissolved Ca. However, biomass regrowth could be observed after CaCO3 precipitation ceased, which suggests that ureolysis-induced CaCO3 precipitation is not necessarily lethal for the entire population. The kinetics of ureolysis and CaCO3 precipitation with S. pasteurii was further analyzed under anaerobic conditions. Rate coefficients obtained in anaerobic environments were comparable to those under aerobic conditions; however, no cell growth was observed under anaerobic conditions with NO−3, SO2−4 or Fe3+ as potential terminal electron acceptors. These data suggest that the initial rates of ureolysis and ureolysis-induced CaCO3 precipitation are not significantly affected by the absence of oxygen but that long-term ureolytic activity might require the addition of suitable electron acceptors. Variations in the ureolytic capabilities and associated rates of CaCO3 precipitation between strains must be fully considered in subsurface engineering strategies that utilize microbial amendments.Item Microbial CaCO3 mineral formation and stability in an experimentally simulated high pressure saline aquifer with supercritical CO2(2013-07) Mitchell, Andrew C.; Phillips, Adrienne J.; Schultz, Logan N.; Parks, Stacy L.; Spangler, Lee H.; Cunningham, Alfred B.; Gerlach, RobinThe use of microbiologically induced mineralization to plug pore spaces is a novel biotechnology to mitigate the potential leakage of geologically sequestered carbon dioxide from preferential leakage pathways. The bacterial hydrolysis of urea (ureolysis) which can induce calcium carbonate precipitation, via a pH increase and the production of carbonate ions, was investigated under conditions that approximate subsurface storage environments, using a unique high pressure (∼7.5 MPa) moderate temperature (32 °C) flow reactor housing a synthetic porous media core. The synthetic core was inoculated with the ureolytic organism Sporosarcina pasteurii and pulse-flow of a urea inclusive saline growth medium was established through the core. The system was gradually pressurized to 7.5 MPa over the first 29 days. Concentrations of NH4+, a by-product of urea hydrolysis, increased in the flow reactor effluent over the first 20 days, and then stabilized at a maximum concentration consistent with the hydrolysis of all the available urea. pH increased over the first 6 days from 7 to 9.1, consistent with buffering by NH4+ ⇔ NH3 + H+. Ureolytic colony forming units were consistently detected in the reactor effluent, indicating a biofilm developed in the high pressure system and maintained viability at pressures up to 7.5 MPa. All available calcium was precipitated as calcite. Calcite precipitates were exposed to dry supercritical CO2 (scCO2), water-saturated scCO2, scCO2-saturated brine, and atmospheric pressure brine. Calcite precipitates were resilient to dry scCO2, but suffered some mass loss in water-saturated scCO2 (mass loss 17 ± 3.6% after 48 h, 36 ± 7.5% after 2 h). Observations in the presence of scCO2 saturated brine were ambiguous due to an artifact associated with the depressurization of the scCO2 saturated brine before sampling. The degassing of pressurized brine resulted in significant abrasion of calcite crystals and resulted in a mass loss of approximately 92 ± 50% after 48 h. However dissolution of calcite crystals in brine at atmospheric pressure, but at the pH of the scCO2 saturated brine, accounted for only approximately 7.8 ± 2.2% of the mass loss over the 48 h period. These data suggest that microbially induced mineralization, with the purpose of reducing the permeability of preferential leakage pathways during the operation of GCS, can occur under high pressure scCO2 injection conditions.Item Microbial oxidation as a methane sink beneath the West Antarctic Ice Sheet(2017-07) Michaud, Alexander B.; Dore, John E.; Achberger, Amanda M.; Christner, Brent C.; Mitchell, Andrew C.; Skidmore, Mark L.; Vick-Majors, Trista J.; Priscu, John C.Aquatic habitats beneath ice masses contain active microbial ecosystems capable of cycling important greenhouse gases, such as methane (CH4). A large methane reservoir is thought to exist beneath the West Antarctic Ice Sheet, but its quantity, source and ultimate fate are poorly understood. For instance, O2 supplied by basal melting should result in conditions favourable for aerobic methane oxidation. Here we use measurements of methane concentrations and stable isotope compositions along with genomic analyses to assess the sources and cycling of methane in Subglacial Lake Whillans (SLW) in West Antarctica. We show that sub-ice-sheet methane is produced through the biological reduction of CO2 using H2. This methane pool is subsequently consumed by aerobic, bacterial methane oxidation at the SLW sediment–water interface. Bacterial oxidation consumes >99% of the methane and represents a significant methane sink, and source of biomass carbon and metabolic energy to the surficial SLW sediments. We conclude that aerobic methanotrophy may mitigate the release of methane to the atmosphere upon subglacial water drainage to ice sheet margins and during periods of deglaciation.Item Microbial sulfur transformations in Subglacial Lake Whillans sediments(2014-11) Purcell, Alicia M.; Mikucki, Jill A.; Achberger, Amanda M.; Alekhina, Irina A.; Barbante, Carlo; Christner, Brent C.; Ghosh, Dhritiman; Michaud, Alexander B.; Mitchell, Andrew C.; Priscu, John C.; Scherer, Reed; Skidmore, Mark L.; Vick-Majors, Trista J.Diverse microbial assemblages inhabit subglacial aquatic environments. While few of these environments have been sampled, data reveal that subglacial organisms gain energy for growth from reduced minerals containing nitrogen, iron, and sulfur. Here we investigate the role of microbially mediated sulfur transformations in sediments from Subglacial Lake Whillans (SLW), Antarctica, by examining key genes involved in dissimilatory sulfur oxidation and reduction. The presence of sulfur transformation genes throughout the top 34 cm of SLW sediments changes with depth. SLW surficial sediments were dominated by genes related to known sulfur-oxidizing chemoautotrophs. Sequences encoding the adenosine-5′-phosphosulfate (APS) reductase gene, involved in both dissimilatory sulfate reduction and sulfur oxidation, were present in all samples and clustered into 16 distinct operational taxonomic units. The majority of APS reductase sequences (74%) clustered with known sulfur oxidizers including those within the “Sideroxydans” and Thiobacillus genera. Reverse-acting dissimilatory sulfite reductase (rDSR) and 16S rRNA gene sequences further support dominance of “Sideroxydans” and Thiobacillus phylotypes in the top 2 cm of SLW sediments. The SLW microbial community has the genetic potential for sulfate reduction which is supported by experimentally measured low rates (1.4 pmol cm-3d-1) of biologically mediated sulfate reduction and the presence of APS reductase and DSR gene sequences related to Desulfobacteraceae and Desulfotomaculum. Our results also infer the presence of sulfur oxidation, which can be a significant energetic pathway for chemosynthetic biosynthesis in SLW sediments. The water in SLW ultimately flows into the Ross Sea where intermediates from subglacial sulfur transformations can influence the flux of solutes to the Southern Ocean.Item Microbially enhanced carbon capture and storage by mineral-trapping and solubility-trapping(2010-07) Mitchell, Andrew C.; Dideriksen, K.; Spangler, Lee H.; Cunningham, Alfred B.; Gerlach, RobinThe potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO2 is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO2)] of 13C-CO2. Dissolved Ca2+ in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L-1 urea. The incorporation of carbonate ions from 13C-CO2 (13C-CO32-) into calcite increased with increasing p(13CO2) and increasing urea concentrations: from 8.3% of total carbon in CaCO3 at 1 g L-1 to 31% at 5 g L-1, and 37% at 20 g L-1. This demonstrated that ureolysis was effective at precipitating initially gaseous [CO2(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO2) demonstrated that no net precipitation of CO2(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO2(g) into the brine. This reduced the headspace concentration of CO2 by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO2, this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO2(g) leakage reduction.Item Microbially enhanced carbonate mineralization and the geologic containment of CO2(2008) Mitchell, Andrew C.; Phillips, Adrienne J.; Kaszuba, John P.; Hollis, W. Kirk; Cunningham, Alfred B.; Gerlach, RobinGeologic sequestration of CO2 involves injection into deep underground formations including oil beds, un-minable coal seams, and saline aquifers with temperature and pressure conditions such that CO2 will likely be in the supercritical state. Supercritical CO2 injection into the receiving formation will result in elevated pressure in the region surrounding the point of injection, and may result in an upward hydrodynamic pressure gradient and associated “leakage†of supercritical to gaseous CO2. Therefore mechanisms to reduce leakage and to mineralize CO2 in a solid form are extremely advantageous for the long-term geologic containment of CO2.This paper will focus on microbially-based strategies for controlling leakage and sequestrating supercritical CO2 during geologic injection. We will examine the concept of using engineered microbial barriers (Cunningham et al., in review; Mitchell et al., in review) which are capable of precipitating calcium carbonate (Mitchell and Ferris, 2005; 2006) under high-pressure subsurface conditions. These “biomineralization barriers†may provide a method for plugging preferential flow pathways in the vicinity of CO2 injection, thereby reducing the potential for unwanted upward migration of CO2, as well as mineralizing injected CO2. A summary of experiments investigating biofilm and associated calcium carbonate formation in porous media using a unique high pressure (8.9 MPa), moderate temperature (≥ 32 °C) flow reactor will be presented, and the potential for biomineralization enhanced CO2 sequestration discussed.Item Microbially enhanced geologic containment of sequestered supercritical CO2(2009-02) Cunningham, Alfred B.; Gerlach, Robin; Spangler, Lee H.; Mitchell, Andrew C.Geologic sequestration of CO2 involves injection into underground formations including oil beds, deep un-minable coal seams, and deep saline aquifers with temperature and pressure conditions such that CO2 will likely be in the supercritical state. It is important that the receiving aquifer have sufficient porosity and permeability and be overlain by a suitable low-permeability cap rock formation. Supercritical CO2 injected into the receiving formation is only slightly soluble in water (approximately 4%) and therefore two fluid phases develop. Also, supercritical CO2 is less dense and much less viscous than the initially resident brine resulting in the potential for upward leakage of CO2 through fractures, disturbed rock, or cement lining near injection wells. This paper summarizes recent research on microbially-based strategies for controlling leakage of CO2 during geologic sequestration. We examine the concept of using engineered microbial biofilms which are capable of precipitating crystalline calcium carbonate using the process of ureolysis. The resulting combination of biofilm plus mineral deposits, if targeted near points of CO2 injection, may result in the long-term sealing of preferential leakage pathways. Successful development of these biologically-based concepts could result in a CO2 leakage mitigation technology which can be applied either before CO2 injection or as a remedial measure.Item Need for direct measurements of coupled microbiological and hydrological processes at different scales in porous media systems(2008-01) Geesey, Gill G.; Mitchell, Andrew C.Reactive transport models contain terms describing microbiological and hydrological processes that control fate and transport of contaminants in porous media. Most models assume that microbial reaction rate is independent of microbial biomass distribution or that biomass is uniformly distributed across media surfaces in a manner that mass transport does not limit reaction rate. Experimental data, as well as some computational models, however, suggest otherwise, indicating a need to experimentally establish how the coupling of microbial biomass and flow distribution influence microbial reaction rates. Nuclear magnetic resonance techniques offer the opportunity to quantify in three dimensions the coupling of microbial biomass and flow velocity distribution in opaque porous media at multiple scales in a noninvasive manner. Experimental data obtained with these techniques can be used to improve the accuracy of boundary conditions used by reactive transport models to predict contaminant fate and transport at the pore and core scales. Further improvements in surface and subsurface magnetic resonance techniques may allow future detection and measurement of microbial biomass distribution in the subsurface at the field scale.Item Physiological Ecology of Microorganisms in Subglacial Lake Whillans(2016-10) Vick-Majors, Trista J.; Mitchell, Andrew C.; Achberger, Amanda M.; Christner, Brent C.; Dore, John E.; Michaud, Alexander B.; Mikucki, Jill A.; Purcell, Alicia M.; Skidmore, Mark L.; Priscu, John C.Subglacial microbial habitats are widespread in glaciated regions of our planet. Some of these environments have been isolated from the atmosphere and from sunlight for many thousands of years. Consequently, ecosystem processes must rely on energy gained from the oxidation of inorganic substrates or detrital organic matter. Subglacial Lake Whillans (SLW) is one of more than 400 subglacial lakes known to exist under the Antarctic ice sheet; however, little is known about microbial physiology and energetics in these systems. When it was sampled through its 800 m thick ice cover in 2013, the SLW water column was shallow (~2 m deep), oxygenated, and possessed sufficient concentrations of C, N, and P substrates to support microbial growth. Here, we use a combination of physiological assays and models to assess the energetics of microbial life in SLW. In general, SLW microorganisms grew slowly in this energy-limited environment. Heterotrophic cellular carbon turnover times, calculated from (3)H-thymidine and (3)H-leucine incorporation rates, were long (60 to 500 days) while cellular doubling times averaged 196 days. Inferred growth rates (average ~0.006 d(-1)) obtained from the same incubations were at least an order of magnitude lower than those measured in Antarctic surface lakes and oligotrophic areas of the ocean. Low growth efficiency (8%) indicated that heterotrophic populations in SLW partition a majority of their carbon demand to cellular maintenance rather than growth. Chemoautotrophic CO2-fixation exceeded heterotrophic organic C-demand by a factor of ~1.5. Aerobic respiratory activity associated with heterotrophic and chemoautotrophic metabolism surpassed the estimated supply of oxygen to SLW, implying that microbial activity could deplete the oxygenated waters, resulting in anoxia. We used thermodynamic calculations to examine the biogeochemical and energetic consequences of environmentally imposed switching between aerobic and anaerobic metabolisms in the SLW water column. Heterotrophic metabolisms utilizing acetate and formate as electron donors yielded less energy than chemolithotrophic metabolisms when calculated in terms of energy density, which supports experimental results that showed chemoautotrophic activity in excess of heterotrophic activity. The microbial communities of subglacial lake ecosystems provide important natural laboratories to study the physiological and biogeochemical behavior of microorganisms inhabiting cold, dark environments.Item Potential CO2 leakage reduction through biofilm-induced calcium carbonate precipitation(2013-01) Phillips, Adrienne J.; Lauchnor, Ellen G.; Eldring, Joseph; Esposito, R.; Mitchell, Andrew C.; Gerlach, Robin; Cunningham, Alfred B.; Spangler, Lee H.Mitigation strategies for sealing high permeability regions in cap rocks, such as fractures or improperly abandoned wells, are important considerations in the long term security of geologically stored carbon dioxide (CO2). Sealing technologies using low-viscosity fluids are advantageous in this context since they potentially reduce the necessary injection pressures and increase the radius of influence around injection wells. Using aqueous solutions and suspensions that can effectively promote microbially induced mineral precipitation is one such technology. Here we describe a strategy to homogenously distribute biofilm-induced calcium carbonate (CaCO3) precipitates in a 61 cm long sandfilled column and to seal a hydraulically fractured, 74 cm diameter Boyles Sandstone core. Sporosarcina pasteurii biofilms were established and an injection strategy developed to optimize CaCO3 precipitation induced via microbial urea hydrolysis. Over the duration of the experiments, permeability decreased between 2 and 4 orders of magnitude in sand column and fractured core experiments, respectively. Additionally, after fracture sealing, the sandstone core withstood three times higher well bore pressure than during the initial fracturing event, which occurred prior to biofilm-induced CaCO3 mineralization. These studies suggest biofilm-induced CaCO3 precipitation technologies may potentially seal and strengthen fractures to mitigate CO2 leakage potential.Item Reducing the risk of well bore leakage using engineered biomineralization barriers(2011-04) Cunningham, Alfred B.; Gerlach, Robin; Spangler, Lee H.; Mitchell, Andrew C.; Parks, Stacy L.; Phillips, Adrienne J.If CO2 is injected in deep geological formations it is important that the receiving formation has sufficient porosity and permeability for storage and transmission and be overlain by a suitable low-permeability cap rock formation. When the resulting CO2 plume encounters a well bore, leakage may occur through various pathways in the “disturbed zone” surrounding the well casing. Gasda et al. , propose a method for determining effective well bore permeability from a field pressure test. If permeability results from such tests prove unacceptably large, strategies for in situ mitigation of potential leakage pathways become important. To be effective, leakage mitigation methods must block leakage pathways on timescales longer than the plume will be mobile, be able to be delivered without causing well screen plugging, and be resistant to supercritical CO2 (ScCO2) challenges. Traditional mitigation uses cement, a viscous fluid that requires a large enough aperture for delivery and that also must bond to the surrounding surfaces in order to be effective. Technologies that can be delivered via low viscosity fluids and that can effectively plug small aperture pathways, or even the porous rock surrounding the well could have significant advantages for some leakage scenarios. We propose a microbially mediated method for plugging preferential leakage pathways and/or porous media, thereby lowering the risk of unwanted upward migration of CO2, similar to that discussed by Mitchell et al. .We examine the concept of using engineered microbial biofilms which are capable of precipitating crystalline calcium carbonate using the process of ureolysis. The resulting combination of biofilm plus mineral deposits, if targeted near points of CO2 injection, may result in the long-term sealing of preferential leakage pathways. Successful development of these biologically-based concepts could result in a CO2 leakage mitigation technology which can be applied either before CO2 injection or as a remedial measure. Results from laboratory column studies are presented which illustrate how biomineralization deposits can be developed along packed sand columns at length scales of 2.54 cm and 61 cm. Strategies for controlling mineral deposition of uniform thickness along the axis of flow are also discussed.