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Item Pharmaceutical impacts on aerobic granular sludge morphology and potential implications for abiotic removal(Elsevier, 2024-02) Bodle, Kylie B.; Kirkland, Catherine M.The goal of this study was to investigate abiotic pharmaceutical removal and abiotic pharmaceutical effects on aerobic granular sludge morphology. For 80 days, a pharmaceutical mixture containing approximately 150 μg/L each of diclofenac, erythromycin, and gemfibrozil was fed to an aerobic granular sludge sequencing batch reactor and granule characteristics were compared with those from a control reactor. Aqueous and solid phase pharmaceutical concentrations were monitored and staining was used to assess changes in biofilm structures. Solid phase pharmaceutical concentrations were elevated over the first 12 days of dosing; however, they then dropped, indicative of desorption. The lipid content in pharmaceutical-exposed granules declined by approximately half over the dosing period, though the relative concentrations of other key biofilm components (proteins, alpha-, and beta-polysaccharides) did not change. Batch experiments were conducted to try to find an explanation for the desorption observed, but reduced solid phase pharmaceutical concentrations could not be linked with the presence of common wastewater constituents such as ammonia or phosphate. Sorption of all three compounds was modeled best by the Henry isotherm, indicating that, even at 150 μg/L, granules’ sorption site coverage was incomplete. Altogether, this study demonstrates that simplified batch systems may not accurately represent the complex abiotic processes occurring in flow-through, biotic systems.Item Treatment performance and microbial community structure in an aerobic granular sludge sequencing batch reactor amended with diclofenac, erythromycin, and gemfibrozil(Frontiers Media SA, 2023-09) Bodle, Kylie B.; Mueller, Rebecca C.; Pernat, Madeline R.; Kirkland, Catherine M.This study characterizes the effects of three commonly detected pharmaceuticals—diclofenac, erythromycin, and gemfibrozil—on aerobic granular sludge. Approximately 150 µg/L of each pharmaceutical was fed in the influent to a sequencing batch reactor for 80 days, and the performance of the test reactor was compared with that of a control reactor. Wastewater treatment efficacy in the test reactor dropped by approximately 30-40%, and ammonia oxidation was particularly inhibited. The relative abundance of active Rhodocyclaceae, Nitrosomonadaceae, and Nitrospiraceae families declined throughout exposure, likely explaining reductions in wastewater treatment performance. Pharmaceuticals were temporarily removed in the first 12 days of the test via both sorption and degradation; both removal processes declined sharply thereafter. This study demonstrates that aerobic granular sludge may successfully remove pharmaceuticals in the short term, but long-term tests are necessary to confirm if pharmaceutical removal is sustainable.Item Pharmaceutical Sorption to Lab Materials May Overestimate Rates of Removal in Lab-Scale Bioreactors(Springer Science and Business Media LLC, 2022-11) Bodle, Kylie B.; Pernat, Madeline R.; Kirkland, Catherine M.Environmental contamination from pharmaceuticals has received increased attention from researchers in the past 20 years. As such, numerous lab-scale studies have sought to characterize the effects of these contaminants on various targets, as well as determine improved removal methods. Many studies have used lab-scale bioreactors to investigate pharmaceutical effects on wastewater bacteria, as wastewater treatment plants often act as reservoirs for pharmaceuticals. However, few—if any—of these studies report the specific lab materials used during testing, such as tubing or pipette tip type. In this study, the pharmaceuticals erythromycin, diclofenac, and gemfibrozil were exposed to different micropipette tips, syringe filters, and tubing types, and losses over time were evaluated. Losses to tubing and syringe filters were particularly significant and neared 100%, depending on the pharmaceutical compound and length of exposure time. Results discussed herein indicate that pharmaceutical sorption to various lab supplies results in decreases to both dosed and quantified pharmaceutical concentrations. Studies that fail to consider this source of loss may therefore draw inaccurate conclusions about pharmaceutical effects or removal efficiencies.Item Addressing wellbore integrity and thief zone permeability using microbially-induced calcium carbonate precipitation (MICP): A field demonstration(Elsevier BV, 2020-02) Kirkland, Catherine M.; Thane, Abby; Hiebert, Randy; Hyatt, Robert; Kirksey, Jim; Cunningham, Alfred B.; Gerlach, Robin; Spangler, Lee; Philips, Adrienne J.Microbially-induced calcium carbonate precipitation (MICP) is an emerging biotechnology for wellbore integrity applications including sealing defects in wellbore cement and modifying the permeability of rock formations. The goal of this field demonstration was to characterize a failed waterflood injection well and provide proof of principle that MICP can reduce permeability in the presence of oil using conventional oilfield fluid delivery methods. We compared well logs performed at the time the well was drilled with ultrasonic logs, sonic cement evaluation, and temperature logs conducted after the well failed. Analysis of these logs suggested that, rather than entering the target waterflood formation, injectate was traveling through defects in the well cement to a higher permeability sandstone layer above the target formation. Sporosarcina pasteurii cultures and urea-calcium media were delivered 2290 ft (698 m) below ground surface using a 3.75 gal (14.2 L) slickline dump bailer to promote mineralization in the undesired flow paths. By Day 6 and after 25 inoculum and 49 calcium media injections, the injectivity [gpm/psi] had decreased by approximately 70%. This demonstration shows that 1) common well logs can be used to identify scenarios where MICP can be employed to reduce system permeability, remediate leakage pathways, and improve waterflood efficiency, and 2) MICP can occur in the presence of hydrocarbons.Item 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.Item 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.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.