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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    Characterizing the structure of aerobic granular sludge using ultra-high field magnetic resonance
    (IWA Publishing, 2020-08) Kirkland, Catherine M.; Krug, Julia R.; Vergeldt, Frank J.; van den Berg, Lenno; Velders, Aldrik H.; Seymour, Joseph D.; Codd, Sarah L.; Van As, Henk; de Kreuk, Merle K.
    Despite aerobic granular sludge wastewater treatment plants operating around the world, our understanding of internal granule structure and its relation to treatment efficiency remains limited. This can be attributed in part to the drawbacks of time-consuming, labor-intensive, and invasive microscopy protocols which effectively restrict samples sizes and may introduce artefacts. Timedomain nuclear magnetic resonance (NMR) allows non-invasive measurements which describe internal structural features of opaque, complex materials like biofilms. NMR was used to image aerobic granules collected from five full-scale wastewater treatment plants in the Netherlands and United States, as well as laboratory granules and control beads. T1 and T2 relaxation-weighted images reveal heterogeneous structures that include high- and low-density biofilm regions, waterlike voids, and solid-like inclusions. Channels larger than approximately 50 μm and connected to the bulk fluid were not visible. Both cluster and ring-like structures were observed with each granule source having a characteristic structural type. These structures, and their NMR relaxation behavior, were stable over several months of storage. These observations reveal the complex structures within aerobic granules from a range of sources and highlight the need for non-invasive characterization methods like NMR to be applied in the ongoing effort to correlate structure and function.
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    Heterogeneous diffusion in aerobic granular sludge
    (Wiley, 2020-08) van den Berg, Lenno; Kirkland, Catherine M.; Seymour, Joseph D.; Codd, Sarah L.; Van Loosdrecht, Mark C. M.; de Kreuk, Merle K.
    Aerobic granular sludge (AGS) technology allows simultaneous nitrogen, phosphorus, and carbon removal in compact wastewater treatment processes. To operate, design, and model AGS reactors, it is essential to properly understand the diffusive transport within the granules. In this study, diffusive mass transfer within full‐scale and lab‐scale AGS was characterized with nuclear magnetic resonance (NMR) methods. Self‐diffusion coefficients of water inside the granules were determined with pulsed‐field gradient NMR, while the granule structure was visualized with NMR imaging. A reaction‐diffusion granule‐scale model was set up to evaluate the impact of heterogeneous diffusion on granule performance. The self‐diffusion coefficient of water in AGS was ∼70% of the self‐diffusion coefficient of free water. There was no significant difference between self‐diffusion in AGS from full‐scale treatment plants and from lab‐scale reactors. The results of the model showed that diffusional heterogeneity did not lead to a major change of flux into the granule (<1%). This study shows that differences between granular sludges and heterogeneity within granules have little impact on the kinetic properties of AGS. Thus, a relatively simple approach is sufficient to describe mass transport by diffusion into the granules.
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    Characterization and quantification of structure and flow in multichannel polymer membranes by MRI
    (2019-01) Schuhmann, S.; Simkins, Jeffrey W.; Schork, N.; Codd, Sarah L.; Seymour, Joseph D.; Heijnen, M.; Saravia, F.; Horn, H.; Nirschl, H.; Guthausen, G.
    Polymeric multichannel hollow fiber membranes were developed to reduce fiber breakage and to increase the volume-to-membrane-surface ratio and consequently the efficiency of filtration processes. These membranes are commonly used in ultrafiltration and are operated in in-out dead-end mode. However, some of the filtration details are unknown. The filtration efficiency and flow in the multichannel membranes depend on filtration time and are expected to vary along spatial coordinates. In the current work, in-situ magnetic resonance imaging was used to answer these questions. Velocities were quantified in the feed channels to obtain a detailed understanding of the filtration process. Flow and deposits were measured in each of the seven channels during filtration of sodium alginate, which is a model substance for extracellular polymeric substances occurring in water treatment. Volume flow and flow profiles were calculated from phase contrast flow images. The flow in z-direction in the center channel was higher than in the surrounding channels. Flow profiles variate depending on the concentration of Ca2+, which changes the filtration mechanism of aqueous solutions of sodium alginate from concentration polarization to gel layer filtration.
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    Spatiotemporal mapping of oxygen in a microbially-impacted packed bed using 19F Nuclear magnetic resonance oximetry
    (2018-08) Simkins, Jeffrey W.; Stewart, Philip S.; Seymour, Joseph D.
    19F magnetic resonance has been used in the medical field for quantifying oxygenation in blood, tissues, and tumors. The 19F NMR oximetry technique exploits the affinity of molecular oxygen for liquid fluorocarbon phases, and the resulting linear dependence of 19F spin–lattice relaxation rate R1 on local oxygen concentration. Bacterial biofilms, aggregates of bacteria encased in a self-secreted matrix of extracellular polymers, are important in environmental, industrial, and clinical settings and oxygen gradients represent a critical determinant of biofilm function. However, measurement of oxygen distribution in biofilms and biofouled porous media is difficult. Here the ability of 19F NMR oximetry to accurately track oxygen profile development in microbial impacted packed bed systems without impacting oxygen transport is demonstrated. Time-stable and inert fluorocarbon containing particles are designed which act as oxygen reporters in porous media systems. Particles are generated by emulsifying and entrapping perfluorooctylbromide (PFOB) into alginate gel, resulting in oxygen-sensing alginate beads that are then used as the solid matrix of the packed bed. 19F oxygenation maps, when combined with 1H velocity maps, allow for insight into the interplay between fluid dynamics and oxygen transport phenomena in these complex biofouled systems. Spatial maps of oxygen consumption rate constants are calculated. The growth characteristics of two bacteria, a non-biofilm forming Escherichia coli and Staphylococcus epidermidis, a strong biofilm-former, are used to demonstrate the novel data provided by the method.
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    NMR Relaxometry to Characterize the Drug Structural Phase in a Porous Construct
    (2018-06) Thrane, Linn W.; Berglund, Emily A.; Wilking, James N.; Vodak, David; Seymour, Joseph D.
    Nuclear magnetic resonance (NMR) frequency spectra and T2 relaxation time measurements, using a high-power radio frequency probe, are shown to characterize the presence of an amorphous drug in a porous silica construct. The results indicate the ability of non-solid-state NMR methods to characterize crystalline and amorphous solid structural phases in drugs. Two-dimensional T1–T2 magnetic relaxation time correlation experiments are shown to monitor the impact of relative humidity on the drug in a porous silica tablet.
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    Quantifying NMR relaxation correlation and exchange in articular cartilage with time domain analysis
    (2018-02) Mailhiot, Sarah E.; Zong, Fangrong; Maneval, James E.; Galvosas, Petrik; Seymour, Joseph D.
    Measured nuclear magnetic resonance (NMR) transverse relaxation data in articular cartilage has been shown to be multi-exponential and correlated to the health of the tissue. The observed relaxation rates are dependent on experimental parameters such as solvent, data acquisition methods, data analysis methods, and alignment to the magnetic field. In this study, we show that diffusive exchange occurs in porcine articular cartilage and impacts the observed relaxation rates in T1-T2 correlation experiments. By using time domain analysis of T2-T2 exchange spectroscopy, the diffusive exchange time can be quantified by measurements that use a single mixing time. Measured characteristic times for exchange are commensurate with T1 in this material and so impacts the observed T1 behavior. The approach used here allows for reliable quantification of NMR relaxation behavior in cartilage in the presence of diffusive fluid exchange between two environments.
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    NMR investigation of water diffusion in different biofilm structures
    (2017-09) Herrling, M. P.; Weisbrodt, Jessica; Kirkland, Catherine M.; Williamson, Nathan H.; Lackner, S.; Codd, Sarah L.; Seymour, Joseph D.; Guthausen, G.; Horn, H.
    Mass transfer in biofilms is determined by diffusion. Different mostly invasive approaches have been used to measure diffusion coefficients in biofilms, however, data on heterogeneous biomass under realistic conditions is still missing. To non-invasively elucidate fluid–structure interactions in complex multispecies biofilms pulsed field gradient-nuclear magnetic resonance (PFG-NMR) was applied to measure the water diffusion in five different types of biomass aggregates: one type of sludge flocs, two types of biofilm, and two types of granules. Data analysis is an important issue when measuring heterogeneous systems and is shown to significantly influence the interpretation and understanding of water diffusion. With respect to numerical reproducibility and physico-chemical interpretation, different data processing methods were explored: (bi)-exponential data analysis and the Γ distribution model. Furthermore, the diffusion coefficient distribution in relation to relaxation was studied by D-T2 maps obtained by 2D inverse Laplace transform (2D ILT). The results show that the effective diffusion coefficients for all biofilm samples ranged from 0.36 to 0.96 relative to that of water. NMR diffusion was linked to biofilm structure (e.g., biomass density, organic and inorganic matter) as observed by magnetic resonance imaging and to traditional biofilm parameters: diffusion was most restricted in granules with compact structures, and fast diffusion was found in heterotrophic biofilms with fluffy structures. The effective diffusion coefficients in the biomass were found to be broadly distributed because of internal biomass heterogeneities, such as gas bubbles, precipitates, and locally changing biofilm densities. Thus, estimations based on biofilm bulk properties in multispecies systems can be overestimated and mean diffusion coefficients might not be sufficiently informative to describe mass transport in biofilms and the near bulk.
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    Peclet number dependent superdiffusive hydrodynamic dispersion in a site percolation porous media measured by NMR
    (2017-04) Seymour, Joseph D.; Codd, Sarah L.; Kimmich, Rainer
    The displacement time dependent hydrodynamic dispersion in a model 2D site percolation structure is measured using PGSE NMR. The data indicate superdiffusive scaling of the mean squared displacement at high Peclet numbers, where advective transport dominates, consistent with classic percolation scaling concepts. The time scaling of the mean squared displacement is shown to vary with the Peclet number demonstrating a dependence on the changing dynamics.
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    Anomalous fluid transport in porous media induced by biofilm growth
    (2004-11) Seymour, Joseph D.; Gage, Justin P.; Codd, Sarah L.; Gerlach, Robin
    Magnetic resonance measurements of the transition from normal to anomalous hydrodynamic dispersion in porous media due to biological activity are presented. Fractional advection-diffusion equations are shown to provide models for the measured impact of biofilm growth on porous media transport dynamics.
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