Scholarly Work - Center for Biofilm Engineering

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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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    Symmetry breaking propulsion of magnetic microspheres in nonlinearly viscoelastic fluids
    (Springer Nature, 2021-02) Rogowski, Louis William; Ali, Jamel; Zhang, Xiao; Wilking, James N.; Fu, Henry C.; Kim, Min Jun
    Microscale propulsion impacts a diverse array of fields ranging from biology and ecology to health applications, such as infection, fertility, drug delivery, and microsurgery. However, propulsion in such viscous drag-dominated fluid environments is highly constrained, with time-reversal and geometric symmetries ruling out entire classes of propulsion. Here, we report the spontaneous symmetry-breaking propulsion of rotating spherical microparticles within non-Newtonian fluids. While symmetry analysis suggests that propulsion is not possible along the fore-aft directions, we demonstrate the existence of two equal and opposite propulsion states along the sphere’s rotation axis. We propose and experimentally corroborate a propulsion mechanism for these spherical microparticles, the simplest microswimmers to date, arising from nonlinear viscoelastic effects in rotating flows similar to the rod-climbing effect. Similar possibilities of spontaneous symmetry-breaking could be used to circumvent other restrictions on propulsion, revising notions of microrobotic design and control, drug delivery, microscale pumping, and locomotion of microorganisms.
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    Coupling fluid flow to hydrogel fluidic devices with reversible “pop-it” connections
    (Royal Society of Chemistry, 2021-01) Abbasi, Reha; LeFevre, Thomas B.; Benjamin, Aaron D.; Thornton, Isaak J.; Wilking, James N.
    Here, we describe a simple, reversible, plug-based connector designed to couple microfluidic tubing to a hydrogel-based fluidic device, to allow for pressurized liquid flow through the system.
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    Physical and chemical mechanisms that influence the electrical conductivity of lignin-derived biochar
    (2021-10) Kane, Seth; Ulrich, Rachel; Harrington, Abigail; Stadie, Nicholas P.; Ryan, Cecily A.
    Lignin-derived biochar is a promising, sustainable alternative to petroleum-based carbon powders (e.g., carbon black) for polymer composite and energy storage applications. Prior studies of these biochars demonstrate that high electrical conductivity and good capacitive behavior are achievable. However, these studies also show high variability in electrical conductivity between biochars (– S/cm). The underlying mechanisms that lead to desirable electrical properties in these lignin-derived biochars are poorly understood. In this work, we examine the causes of the variation in conductivity of lignin-derived biochar to optimize the electrical conductivity of lignin-derived biochars. To this end, we produced biochar from three different lignins, a whole biomass source (wheat stem), and cellulose at two pyrolysis temperatures (900 °C, 1100 °C). These biochars have a similar range of conductivities (0.002 to 18.51 S/cm) to what has been reported in the literature. Results from examining the relationship between chemical and physical biochar properties and electrical conductivity indicate that decreases in oxygen content and changes in particle size are associated with increases in electrical conductivity. Importantly, high variation in electrical conductivity is seen between biochars produced from lignins isolated with similar processes, demonstrating the importance of the lignin’s properties on biochar electrical conductivity. These findings indicate how lignin composition and processing may be further selected and optimized to target specific applications of lignin-derived biochars.
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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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    Light-Based 3D Printing of Hydrogels with High-Resolution Channels
    (2019-01) Benjamin, Aaron D.; Abbasi, Reha; Owens, Madison; Olsen, Robert J.; Walsh, Danica J.; LeFevre, Thomas B.; Wilking, James N.
    Hydrogels are soft, water-based gels with widespread applications in personal care products, medicine and biomedical engineering. Many applications require structuring the hydrogel into complex three-dimensional (3D) shapes. For these applications, light-based 3D printing methods offer exquisite control over material structure. However, the use of these methods for structuring hydrogels is underdeveloped. In particular, the ability to print hydrogel objects containing internal voids and channels is limited by the lack of well-characterized formulations that strongly attenuate light and the lack of a theoretical framework for predicting and mitigating channel occlusion. Here we present a combined experimental and theoretical approach for creating well-defined channels with any orientation in hydrogels using light-based 3D printing. This is achieved by the incorporation of photoblocker and the optimization of print conditions to ensure layer-layer adhesion while minimizing channel occlusion. To demonstrate the value of this approach we print hydrogels containing individual spiral channels with centimeter-scale length and submillimeter-scale cross-section. While the channels presented here are relatively simple, this same approach could be used to achieve more complex channel designs mimicking, for example, the complex vasculature of living organisms. The low cytotoxicity of the gel makes the formulation a promising candidate for biological applications.
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    Biopolymer and water dynamics in microbial biofilm extracellular polymeric substance
    (2008-09) Hornemann, Jennifer A.; Lysova, Anna A.; Codd, Sarah L.; Seymour, Joseph D.; Busse, S.; Stewart, Philip S.; Brown, Jennifer R.
    Nuclear magnetic resonance (NMR) is a noninvasive and nondestructive tool able to access several observable quantities in biofilms such as chemical composition, diffusion, and macroscale structure and transport. Pulsed gradient spin echo (PGSE) NMR techniques were used to measure spectrally resolved biomacromolecular diffusion in biofilm biomass, extending previous research on spectrally resolved diffusion in biofilms. The dominant free water signal was nulled using an inversion recovery modification of the traditional PGSE technique in which the signal from free water is minimized in order to view the spectra of components such as the rotationally mobile carbohydrates, DNA, and proteins. Diffusion data for the major constituents obtained from each of these spectral peaks demonstrate that the biomass of the biofilm contains both a fast and slow diffusion component. The dependence of diffusion on antimicrobial and environmental challenges suggests the polymer molecular dynamics measured by NMR are a sensitive indicator of biofilm function.
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    Escherichia coli O157:H7 attachment and persistence within root biofilm of common treatment wetlands plants
    (2017-01) VanKempen-Fryling, Rachel J.; Camper, Anne K.
    Pathogen retention and subsequent release within the rhizosphere of wastewater treatment wetlands may be a concern for human health. To address this concern, the enteric pathogen Escherichia coli O157:H7 with a DsRed plasmid insertion was used as a model pathogenic organism in an open-air chemostat reactor with constant flow of simulated wastewater. Colonization and persistence of the organism was tracked on roots of two obligate wetland plant species, Carex utriculata and Schoenoplectus acutus, originally grown in pilot scale wetland reactors. Teflon nylon string, clean and with existing indigenous biofilm, was used as an inert surface control. Epifluorescence microscopy and qPCR were used to verify E. coli O157:H7 abundance for up to 1 week. Initial attachment was seen on all surfaces, with colonization decreasing through 1 week. qPCR showed preferential association of the pathogen with roots over the nylon. There was a significant difference between plant type; S. acutus showed significantly higher numbers compared to C. utriculata. E. coli O157:H7 binding and persistence on root surfaces may be a means of survival in treatment wetlands.
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    Nitrification and potential control mechanisms in simulated premises plumbing
    (2011-11) Rahman, Mohammad S.; Encarnacion, G.; Camper, Anne K.
    Indigenous drinking water organisms were used to establish nitrification in glass reactors containing copper or polyvinyl chloride (PVC) surfaces. The reactors were fed soil-derived humics as the organic carbon source and ammonium sulfate as the nitrogen source in biologically treated tap water. Water in the reactors was stagnant for 8 h and then flowed for 5 min to simulate conditions in household plumbing. Following the establishment of complete nitrification (conversion of ammonia to nitrate) in both reactor types, various inhibitors of nitrification were tested followed by a period where recovery of nitrification was observed. In one PVC reactor, copper was gradually introduced up to 1.3 ppm. To ensure that most of the copper was in the ionic form, the pH of the influent was then gradually lowered to 6.6. No significant change in nitrification was observed in the presence of copper. Chlorite was introduced into copper and PVC reactors at doses increasing from 0.2 ppm to 20 ppm. There was limited effect on the PVC system and inhibition in the copper reactor only at 20 ppm. Chloramine was tested at chlorine to ammonia ratios ranging from 0.5:1 to 5:1. Nitrification activity was impacted significantly at a 5:1 ratio and ultimately stopped, with the fastest response being in the copper system. Whenever a control mechanism was tested, there was increased release of copper from the reactors with copper coupons. In all cases, nitrification recovered when inhibitors were removed but the rates of recovery differed depending on the treatment method and coupon surface.
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    New methods for the detection of orthopedic and other biofilm infections
    (2011-03) Costerton, J. William; Post, J. C.; Ehrlich, Garth D.; Hu, Fen Z.; Kreft, R.; Nistico, L.; Kathju, S.; Stoodley, Paul; Hall-Stoodley, Luanne; Maale, G.; James, Garth A.; Sotereanos, N.; DeMeo, P.
    The detection and identification of bacteria present in natural and industrial ecosystems is now entirely based on molecular systems that detect microbial RNA or DNA. Culture methods were abandoned in the 1980s because direct observations showed that <1% of the bacteria in these systems grew on laboratory media. Culture methods comprise the backbone of the Food and Drug Administration-approved diagnostic systems used in hospital laboratories, with some molecular methods being approved for the detection of specific pathogens that are difficult to grow in vitro. In several medical specialties, the reaction to negative cultures in cases in which overt signs of infection clearly exist has produced a spreading skepticism concerning the sensitivity and accuracy of traditional culture methods.We summarize evidence from the field of orthopedic surgery, and from other medical specialties, that support the contention that culture techniques are especially insensitive and inaccurate in the detection of chronic biofilm infections. We examine the plethora of molecular techniques that could replace cultures in the diagnosis of bacterial diseases, and we identify the new Ibis technique that is based on base ratios (not base sequences), as the molecular system most likely to fulfill the requirements of routine diagnosis in orthopedic surgery.
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