Center for Biofilm Engineering (CBE)

Permanent URI for this communityhttps://scholarworks.montana.edu/handle/1/9334

At the Center for Biofilm Engineering (CBE), multidisciplinary research teams develop beneficial uses for microbial biofilms and find solutions to industrially relevant biofilm problems. The CBE was established at Montana State University, Bozeman, in 1990 as a National Science Foundation Engineering Research Center. As part of the MSU College of Engineering, the CBE gives students a chance to get a head start on their careers by working on research teams led by world-recognized leaders in the biofilm field.

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    Beyond the Surface: Non-Invasive Low-Field NMR Analysis of Microbially-Induced Calcium Carbonate Precipitation in Shale Fractures
    (Springer Science and Business Media LLC, 2024-07) Willet, Matthew R.; Bedey, Kayla; Crandall, Dustin; Seymour, Joseph D.; Rutqvist, Jonny; Cunningham, Alfred B.; Phillips, Adrienne J.; Kirkland, Catherine M.
    Microbially-induced calcium carbonate precipitation (MICP) is a biological process in which microbially-produced urease enzymes convert urea and calcium into solid calcium carbonate (CaCO3) deposits. MICP has been demonstrated to reduce permeability in shale fractures under elevated pressures, raising the possibility of applying this technology to enhance shale reservoir storage safety. For this and other applications to become a reality, non-invasive tools are needed to determine how effectively MICP seals shale fractures at subsurface temperatures. In this study, two different MICP strategies were tested on 2.54 cm diameter and 5.08 cm long shale cores with a single fracture at 60 ℃. Flow-through, pulsed-flow MICP-treatment was repeatedly applied to Marcellus shale fractures with and without sand (“proppant”) until reaching approximately four orders of magnitude reduction in apparent permeability, while a single application of polymer-based “immersion” MICP-treatment was applied to an Eagle Ford shale fracture with proppant. Low-field nuclear magnetic resonance (LF-NMR) and X-Ray computed microtomography (micro-CT) techniques were used to assess the degree of biomineralization. With the flow-through approach, these tools revealed that while CaCO3 precipitation occurred throughout the fracture, there was preferential precipitation around proppant. Without proppant, the same approach led to premature sealing at the inlet side of the core. In contrast, immersion MICP-treatment sealed off the fracture edges and showed less mineral precipitation overall. This study highlights the use of LF-NMR relaxometry in characterizing fracture sealing and can help guide NMR logging tools in subsurface remediation efforts.
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    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.
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    A Numerical Model for Enzymatically Induced Calcium Carbonate Precipitation
    (MDPI, 2020-06) Hommel, Johannes; Akyel, Arda; Frieling, Zachary; Phillips, Adrienne J.; Gerlach, Robin; Cunningham, Alfred B.; Class, Holger
    Enzymatically induced calcium carbonate precipitation (EICP) is an emerging engineered mineralization method similar to others such as microbially induced calcium carbonate precipitation (MICP). EICP is advantageous compared to MICP as the enzyme is still active at conditions where microbes, e.g., Sporosarcina pasteurii, commonly used for MICP, cannot grow. Especially, EICP expands the applicability of ureolysis-induced calcium carbonate mineral precipitation to higher temperatures, enabling its use in leakage mitigation deeper in the subsurface than previously thought to be possible with MICP. A new conceptual and numerical model for EICP is presented. The model was calibrated and validated using quasi-1D column experiments designed to provide the necessary data for model calibration and can now be used to assess the potential of EICP applications for leakage mitigation and other subsurface modifications.
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    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 models
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    Biomineralization of Plastic Waste to Improve the Strength of Plastic-Reinforced Cement Mortar
    (2021-04) Kane, Seth; Thane, Abby; Espinal, Michael; Lunday, Kendra; Armagan, Hakan; Phillips, Adrienne J.; Heveran, Chelsea M.; Ryan, Cecily A.
    The development of methods to reuse large volumes of plastic waste is essential to curb the environmental impact of plastic pollution. Plastic-reinforced cementitious materials (PRCs), such as plastic-reinforced mortar (PRM), may be potential avenues to productively use large quantities of low-value plastic waste. However, poor bonding between the plastic and cement matrix reduces the strength of PRCs, limiting its viable applications. In this study, calcium carbonate biomineralization techniques were applied to coat plastic waste and improved the compressive strength of PRM. Two biomineralization treatments were examined: enzymatically induced calcium carbonate precipitation (EICP) and microbially induced calcium carbonate precipitation (MICP). MICP treatment of polyethylene terephthalate (PET) resulted in PRMs with compressive strengths similar to that of plastic-free mortar and higher than the compressive strengths of PRMs with untreated or EICP-treated PET. Based on the results of this study, MICP was used to treat hard-to-recycle types 3–7 plastic waste. No plastics investigated in this study inhibited the MICP process. PRM samples with 5% MICP-treated polyvinyl chloride (PVC) and mixed type 3–7 plastic had compressive strengths similar to plastic-free mortar. These results indicate that MICP treatment can improve PRM strength and that MICP-treated PRM shows promise as a method to reuse plastic waste.
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    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, Robin
    The 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.
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    Field-scale modeling of microbially induced calcite precipitation
    (2018-11) Cunningham, Alfred B.; Class, Holger; Ebigbo, Anozie; Gerlach, Robin; Phillips, Adrienne J.; Hommel, Johannes
    The biogeochemical process known as microbially induced calcite precipitation (MICP) is being investigated for engineering and material science applications. To model MICP process behavior in porous media, computational simulators must couple flow, transport, and relevant biogeochemical reactions. Changes in media porosity and permeability due to biomass growth and calcite precipitation, as well as their effects on one another must be considered. A comprehensive Darcy-scale model has been developed by Ebigbo et al. (Water Resour. Res. 48(7), W07519, 2012) and Hommel et al. (Water Resour. Res. 51, 3695–3715, 2015) and validated at different scales of observation using laboratory experimental systems at the Center for Biofilm Engineering (CBE), Montana State University (MSU). This investigation clearly demonstrates that a close synergy between laboratory experimentation at different scales and corresponding simulation model development is necessary to advance MICP application to the field scale. Ultimately, model predictions of MICP sealing of a fractured sandstone formation, located 340.8 m below ground surface, were made and compared with corresponding field observations. Modeling MICP at the field scale poses special challenges, including choosing a reasonable model-domain size, initial and boundary conditions, and determining the initial distribution of porosity and permeability. In the presented study, model predictions of deposited calcite volume agree favorably with corresponding field observations of increased injection pressure during the MICP fracture sealing test in the field. Results indicate that the current status of our MICP model now allows its use for further subsurface engineering applications, including well-bore cement sealing and certain fracture-related applications in unconventional oil and gas production.
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    Enhancing wellbore cement integrity with microbially induced calcite precipitation (MICP): A field scale demonstration
    (2018-12) Phillips, Adrienne J.; Troyer, E.; Hiebert, R.; Kirkland, Catherine M.; Gerlach, Robin; Cunningham, Alfred B.; Spangler, Lee H.; Kirksey, J.; Rowe, W.; Esposito, R.
    The presence of delaminations, apertures, fractures, voids and other unrestricted flow channels in the wellbore environment substantially reduces wellbore integrity. Compromised cement may cause a loss of zonal isolation leading to deleterious flow of fluids between zones or to the surface with multiple potential negative impacts including: loss of resource production, reduction of sweep efficiency in EOR operations, and regulatory non-compliance. One potential solution to enhance wellbore integrity is microbially induced calcite precipitation (MICP) to plug preferential flow pathways. MICP is promoted with micrometer-sized organisms and low viscosity (aqueous) solutions thereby facilitating fluid transport into small aperture, potentially tortuous leakage flow paths within the cement column. In this study, MICP treatment of compromised wellbore cement was demonstrated at a depth interval of 310.0–310.57 m (1017–1019 feet) below ground surface (bgs) using conventional oil field subsurface fluid delivery technologies (packer, tubing string, and a slickline deployed bailer). After 25 urea/calcium solution and 10 microbial (Sporosarcina pasteurii) suspension injections, injectivity was reduced from the initial 0.29 cubic meters per hour (m3/h) (1.28 gallons per minute (gpm)) to less than 0.011 m3/h (0.05 gpm). The flow rate was decreased while maintaining surface pumping pressure below a maximum pressure of 81.6 bar (1200 psi) to minimize the potential for fracturing a shale formation dominant in this interval. The pressure decay immediately after each injection ceased decreased after MICP treatment. Comparison of pre- and post-test cement evaluation logs revealed substantial deposition of precipitated solids along the original flow channel. This study suggests MICP is a promising tool for enhancing wellbore cement integrity.
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    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.
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    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, Robin
    Geologic 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.
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