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Item In Situ Enhancement and Isotopic Labeling of Biogenic Coalbed Methane(American Chemical Society, 2022-02) Barnhart, Elliott P.; Ruppert, Leslie; Hiebert, Randy; Smith, Heidi J.; Schweitzer, Hannah D.; Clark, Arthur C.; Weeks, Edwin P.; Orem, William H.; Varonka, Matthew S.; Platt, George; Shelton, Jenna L.; Davis, Katherine J.; Hyatt, Robert J.; McIntosh, Jennifer C.; Ashley, Kilian; Ono, Shuhei; Martini, Anna M.; Hackley, Keith C.; Gerlach, Robin; Spangler, Lee; Phillips, Adrienne J.; Barry, Mark; Cunningham, Alfred B.; Fields, Matthew W.Subsurface microbial (biogenic) methane production is an important part of the global carbon cycle that has resulted in natural gas accumulations in many coal beds worldwide. Laboratory studies suggest that complex carbon-containing nutrients (e.g., yeast or algae extract) can stimulate methane production, yet the effectiveness of these nutrients within coal beds is unknown. Here, we use downhole monitoring methods in combination with deuterated water (D2O) and a 200-liter injection of 0.1% yeast extract (YE) to stimulate and isotopically label newly generated methane. A total dissolved gas pressure sensor enabled real time gas measurements (641 days preinjection and for 478 days postinjection). Downhole samples, collected with subsurface environmental samplers, indicate that methane increased 132% above preinjection levels based on isotopic labeling from D2O, 108% based on pressure readings, and 183% based on methane measurements 266 days postinjection. Demonstrating that YE enhances biogenic coalbed methane production in situ using multiple novel measurement methods has immediate implications for other field-scale biogenic methane investigations, including in situ methods to detect and track microbial activities related to the methanogenic turnover of recalcitrant carbon in the subsurface.Item Temperature‐dependent inactivation and catalysis rates of plant‐based ureases for engineered biomineralization(Wiley, 2020-11) Feder, Marnie J.; Akyel, Arda; Morasko, Vincent J.; Gerlach, Robin; Phillips, Adrienne J.Engineered (bio)mineralization uses the enzyme urease to catalyze the hydrolysis of urea to promote carbonate mineral precipitation. The current study investigates the influence of temperature on ureolysis rate and degree of inactivation of plant-sourced ureases over a range of environmentally relevant temperatures. Batch experiments at 30◦C demonstrated that jack bean meal (JBM) has a 1.7 to 56 times higher activity (844 μmol urea hydrolyzed g−1 JBM min−1) than the other tested plant-sourced ureases (soybean, pigeon pea and cottonseed). Hence, ureolysis and enzyme inactivation rates were evaluated for JBM at temperatures between 20◦C and 80◦C. A combined first-order urea hydrolysis and first-order enzyme inactivation model described the inactivation of urease over the investigated range of temperatures. The temperature-dependent rate coefficients (kurea) increased with temperature and ranged from 0.0018 at 20◦C to 0.0249 L g−1 JBM min−1 at 80◦C; JBM urease became ≥50% inactivated in as little as 5.2 minutes at 80◦C and in as long as 2238 minutes at 50◦C. The combined urea hydrolysis kinetics and enzyme inactivation model provides a mathematical relationship useful for the design of biomineralization technologies and can be incorporated into reactive transport modelsItem 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 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 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 Resilience of planktonic and biofilm cultures to supercritical CO2(2008-12) Mitchell, Andrew C.; Phillips, Adrienne J.; Hamilton, Martin A.; Gerlach, Robin; Hollis, W. Kirk; Kaszuba, John P.; Cunningham, Alfred B.Supercritical CO2 has been shown to act as a disinfectant against microorganisms. These organisms have most often been tested in vegetative or spore form. Since biofilm organisms are typically more resilient to physical, chemical, and biological stresses than the same organisms in planktonic form, they are often considered more difficult to eradicate. It is therefore hypothesized that supercritical CO2 (SC–CO2) induced inactivation of biofilm organisms would be less effective than against planktonic (suspended) growth cultures of the same organism. Six-day old biofilm cultures as well as suspended planktonic cultures of Bacillus mojavensis were exposed to flowing SC–CO2 at 136 atm and 35 ◦C for 19 min and slowly depressurized after treatment. After SC–CO2 exposure, B. mojavensis samples were analyzed for total and viable cells. Suspended cultures revealed a 3 log10 reduction while biofilm cultures showed a 1 log10 reduction in viable cell numbers. These data demonstrate that biofilm cultures of B. mojavensis are more resilient to SC–CO2 than suspended planktonic communities. It is hypothesized that the small reduction in the viability of biofilm microorganisms reflects the protective effects of extracellular polymeric substances (EPS) which make up the biofilm matrix, which offer mass transport resistance, a large surface area, and a number of functional groups for interaction with and immobilization of CO2. The resistance of biofilm suggests that higher pressures, longer durations of SC–CO2 exposure, and a quicker depressurization rate may be required to eradicate biofilms during the sterilization of heat-sensitive materials in medical and industrial applications. However, the observed resilience of biofilms to SC–CO2 is particularly promising for the prospective application of subsurface biofilms in the subsurface geologic sequestration of CO2.Item Detecting Microbially Induced Calcite Precipitation in a Model Well-Bore Using Downhole Low-Field NMR(2017-02) Kirkland, Catherine M.; Zanetti, Sam; Grunewald, Elliot; Walsh, David O.; Codd, Sarah L.; Phillips, Adrienne J.Microbially induced calcite precipitation (MICP) has been widely researched recently due to its relevance for subsurface engineering applications including sealing leakage pathways and permeability modification. These applications of MICP are inherently difficult to monitor nondestructively in time and space. Nuclear magnetic resonance (NMR) can characterize the pore size distributions, porosity, and permeability of subsurface formations. This investigation used a low-field NMR well-logging probe to monitor MICP in a sand-filled bioreactor, measuring NMR signal amplitude and T2 relaxation over an 8 day experimental period. Following inoculation with the ureolytic bacteria, Sporosarcina pasteurii, and pulsed injections of urea and calcium substrate, the NMR measured water content in the reactor decreased to 76% of its initial value. T2 relaxation distributions bifurcated from a single mode centered about approximately 650 ms into a fast decaying population (T2 less than 10 ms) and a larger population with T2 greater than 1000 ms. The combination of changes in pore volume and surface minerology accounts for the changes in the T2 distributions. Destructive sampling confirmed final porosity was approximately 88% of the original value. These results indicate the low-field NMR well-logging probe is sensitive to the physical and chemical changes caused by MICP in a laboratory bioreactor.Item NMR relaxation measurements of biofouling in model and geological porous media(2011-09) Codd, Sarah L.; Vogt, Sarah J.; Hornemann, Jennifer A.; Phillips, Adrienne J.; Maneval, James E.; Romanenko, K. R.; Hansen, L.; Cunningham, Alfred B.; Seymour, Joseph D.Recently 2D nuclear magnetic resonance (NMR) relaxation techniques have been able to access changes in pore structures through surface and diffusion based relaxation measurements. This research investigates the applicability of these methods for measuring pore and surface changes due to biofilm growth in various model porous systems and natural geological media. Model bead packs of various construction containing 100 lm borosilicate and soda lime glass beads were used to demonstrate how changes in the measured relaxation rates can be used to non-invasively verify and quantify biofilm growth in porous media. However significant challenges are shown to arise when trying to implement the same techniques to verify biofilm growth in a natural geological media.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.; Park, Saehan; Phillips, Adrienne J.If CO2 is injected in deep geological formations it is important that the receiving formation hassufficient 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.[9], 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 thatdiscussed by Mitchell et al.[12].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.Item Darcy-scale modeling of microbially induced carbonate mineral precipitation in sand columns(2012-07) Ebigbo, Anozie; Phillips, Adrienne J.; Gerlach, Robin; Helmig, Rainer; Cunningham, Alfred B.; Class, Holger; Spangler, Lee H.This investigation focuses on the use of microbially induced calcium carbonate precipitation (MICP) to set up subsurface hydraulic barriers to potentially increase storage security near wellbores of CO2 storage sites. A numerical model is developed, capable of accounting for carbonate precipitation due to ureolytic bacterial activity as well as the flow of two fluid phases in the subsurface. The model is compared to experiments involving saturated flow through sand-packed columns to understand and optimize the processes involved as well as to validate the numerical model. It is then used to predict the effect of dense-phase CO2 and CO2-saturated water on carbonate precipitates in a porous medium.