Browsing by Subject "Biofilm"

Now showing 1 - 11 of 11
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    Experimental data of biofilm development experiments under fluctuating flow conditions taken and processed at SAFL in 2022
    (2023-05-04) Wei, Guanju; Yang, Judy Q; judyyang@umn.edu; Yang, Judy; University of Minnesota, Saint Anthony Falls Laboratory, Environmental Transport Lab
    This dataset consists of the Matlab codes, experimental data, and raw images of the biofilm development experiments under fluctuating flow conditions. They are all collected in the Saint Anthony Falls Laboratory at the University of Minnesota. The codes are used for processing the raw images to calculate the biofilm thickness and biofilm area coverage. The experimental data file contains the data after processing. The image file contains the raw images collected during the experiments using the Nikon confocal microscope.
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    Experimental data of Pseudomonas putida biofilm development experiments in flat and rough microfluidic channels
    (2022-05-09) Wei, Guanju; Yang, Judy Q; judyyang@umn.edu; Yang, Judy; University of Minnesota, Saint Anthony Falls Laboratory, Environmental Transport Lab
    This dataset consists of the codes, experimental data, and raw images of the Pseudomonas putida biofilm development experiments. They are all collected in the Saint Anthony Falls Laboratory at the University of Minnesota. The codes are used for processing the raw images to calculate the biofilm thickness. The experimental data file contains the experiment parameters and the data after processing. The image file contains the raw images collected using the Nikon confocal microscope.
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    Fate and impact of antibiotics in slow-rate biofiltration processes.
    (2010-12) Wunder, David Barnes
    Antibiotics have been detected in surface waters worldwide at concentrations up to 1.9 micrograms/L, but are typically detected at low nanogram/L concentrations. The potential health effects of exposure to low levels of these compounds via tap water are not known, but there is significant concern among water consumers regarding the occurrence of antibiotics and other pharmaceutical compounds in water supplies. Thus, a significant amount of research has been performed recently to investigate the removal of pharmaceuticals via conventional and advanced water treatment processes. While conventional treatment processes (i.e., coagulation, flocculation, sedimentation, and filtration) are generally not effective, oxidation processes (e.g., chlorination, ozonation) and granular activated carbon exhibit some effectiveness at removing pharmaceuticals. As expected, removals are highly dependent on compound structure. Furthermore, some oxidants, such as chloramines, are not effective at oxidizing pharmaceuticals. Slow-rate biofiltration processes (SRBF), such as slow sand filtration (SSF) and riverbank filtration (RBF), are drinking water treatmeant systems comprised of two stages in sequence: 1) a relatively shallow biotic region where media (i.e., filter sand or aquifer material) is colonized by biofilm bacteria, followed by 2) an deeper abiotic filtration zone. These processes are extensively used in Europe and developing global regions and are seeing increased usage in the United States. There is evidence in the literature that SRBFs can remove a wide variety of trace organic pollutants including: pesticides, disinfection byproducts, and some pharmaceuticals. Little is known regarding the ability of SRBF processes to remove antibiotics from water supplies nor has any work been done to investigate the potential adverse effects of antibiotics on the biofilm bacteria that are critical to SRBF system performance. Thus, this research was performed to determine the extent and mechanisms (i.e., sorption versus biodegradation) of antibiotic removal in SRBF processes and the effects of antibiotics on biofilm bacteria (i.e., activity and community composition). The effect of antibiotics on bacterial activity and community structure was investigated by growing biofilm in the presence and absence of a mixture of antibiotics in a continuous-flow rotating annular bioreactor (CFRAB) with acetate as substrate. Three representative compounds were selected for use in this research: sulfamethoxazole (SMX), erythromycin (ERY), and ciprofloxacin (CIP). These antibiotics were selected because they: 1) represent three prominent classes of antibiotics with differing mechanisms of action against bacteria, 2) have been detected in surface water, 3) exhibit different chemical characteristics, and 4) have differing levels of biodegradability. Areal acetate utilization rates for a constant feed of antibiotics were similar to the control experiments, and utilization rates did not change during an antibiotic shock loading experiment. Attached biomass levels were greater for experiments involving a high CIP concentration (3.33 micrograms/L), however, yielding comparatively lower steady-state biomass-normalized substrate utilization rates. Microbial community analyses via automated ribosomal intergenic spacer analysis (ARISA) revealed shifts in community structure for the high dose CIP experiments. A CFRAB was also used to investigate antibiotic sorption to bacterial biofilm. The extent of sorption, as indicated by the organic carbon partition coefficient (Koc), was 15 to 23 times greater for CIP compared to ERY and SMX. The Koc values did not correlate with experimentally-determined Kow values, suggesting that the sorption of relatively hydrophilic (i.e. Kow < 1.7) and charged antibiotics to typically negatively charged biofilm is driven by ionic interactions (i.e. ion exchange) rather than hydrophobic interactions. The attenuation and impact of antibiotics in SRBF systems was investigated by conducting bench-scale filter column experiments with mixtures of SMX, ERY, and CIP at high (3.33 micrograms/L, each) and low (0.33 microgram/L, each) antibiotic feed conditions. Consistent with the CFRAB experiments, antibiotic breakthrough times were greatest for CIP, with very little uptake of SMX or ERY. Biodegradation was not observed for any antibiotic during 6-weeks of filter column operation or in complementary batch experiments. A one-dimensional advection-dispersion equation (with linear sorption) model was validated against experimental results and used to compare antibiotic retardation in SSF, RBF, and rapid gravity biofiltration (RGBF) systems. Of the modeled systems, antibiotic retardation was greatest in RBF, with little antibiotic removal expected for SSF. Based on analysis of ARISA data, the community structure of bacterial biofilm was not affected in filters exposed to antibiotics at low concentrations (i.e. 0.33 microgram/L, each) similar to those found in surface waters, with a few species impacted under high concentration conditions (3.33 microgram/L, each). The results of this work will help those interested in understanding and predicting antibiotic fate in engineered and natural systems where biofilm is present. The results indicate that antibiotic removal in SRBF processes will be dictated by compound properties such as charge and hydrophobicity, and that limited removal of antibiotics in SRBF processes can be expected. Finally, the results suggest that that mixtures of antibiotics at concentrations typically observed in surface waters are unlikely to adversely affect SRBF biofilm bacteria or process performance.
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    Fluoride and Gallein Inhibit Polyphosphate Accumulation by Oral Pathogen Rothia dentocariosa - Data Sharing Archive
    (2023-01-25) Kumar, Dhiraj; Mandal, Subhrangshu; Bailey, Jake V.; Flood, Beverly E.; Jones, Robert S.; rsjones@umn.edu; Jones, Robert, S; Earth and Environmental Science; School of Dentistry
    This raw data set supports publication found in Letters in Applied Microbiology: The uptake and storage of extracellular orthophosphate (Pi) by polyphosphate (polyP) accumulating bacteria may contribute to mineral dissolution in the oral cavity. To test the effect of potential inhibitors of polyP kinases on Rothia dentocariosa, gallein (0, 25, 50, 100 µM) and fluoride (0, 50, 100 ppm) were added to R. dentocariosa cultures grown in brain heart infusion broth. At late log growth phase (8h), extracellular Pi was measured using an ascorbic acid assay, and polyP was isolated from bacterial cells treated with RNA/DNAases using a neutral phenol/chloroform extraction. Extracts were hydrolyzed and quantified as above. Gallein and fluoride had minor effects on bacterial growth with NaF having a direct effect on media pH. Gallein (≥25 µM) and fluoride (≥50 ppm) attenuated the bacterial drawdown of extracellular Pi 56.7% (p <0.05) and 37.3% (p <0.01). There was a corresponding polyP synthesis decrease of 73.2% (p<0.0001) from gallein and 83.1% (p<0.0001) from fluoride. Attenuated total reflectance Fourier transform infrared spectroscopy validated the presence of polyP and its reduced concentration in R. dentocariosa bacterial cells following gallein and fluoride treatment. R. dentocariosa can directly change extracellular Pi and accumulate intracellular polyP but the mechanism is attenuated by gallein and NaF.
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    Investigating Bi-directional Impacts of the Microbiome and Drinking Water Quality in Drinking Water Distribution System Water Mains and Storage Facilities
    (2022-07) Gomez, Christa Kimloi
    The microbial communities that live in the biofilms of drinking water distribution system (DWDS) environments can exert significant impacts on drinking water quality before it reaches the consumer. The relatively recent advent and accessibility of powerful culture-independent techniques, such as high-throughput sequencing, have enabled characterization of diverse microbial communities; however, the difficulties of accessing DWDS infrastructure has hindered many efforts to study the health-relevant DWDS microbiome. In this work, high-throughput sequencing and quantitative real-time polymerase chain reaction (PCR) techniques were leveraged to characterize the biofilm communities of simulated and full-scale water mains, as well as the in situ suspended and biofilm communities of elevated water storage towers and underground reservoirs in a chloraminated DWDS. Seasonal variability and drivers of community composition were assessed in the simulated DWDS biofilms and in full-scale drinking water storage facilities. Among other examined drivers of community, the presence and concentration of disinfectant was an important selective pressure that impacted community composition. Communities in the simulated and full-scale DWDS biofilms were generally dominated by bacteria that live preferentially in, and form biofilms, exhibit increased resistance to disinfectant concentrations, or display versatility in substrate-utilization. These included genera that contain opportunistic pathogens, such as Mycobacterium, Pseudomonas, and Stenotrophomonas, genera implicated in microbiologically-caused corrosion of infrastructure (sulfate-reducing Desulfovibrio), - as well as ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) that catalyze nitrification processes and can cause reoccurring, problematic decreases in chloramine residual concentrations. Species-level taxonomic resolution of DWDS Mycobacterium, achieved by sequencing a mycobacterial heat shock protein gene, indicated that the bulk of these bacteria were not disease-associated strains. Early stages of community succession occurred rapidly for biofilms on new surfaces that were in proximity to more established biofilms – within a month, the biofilms on new surfaces exhibited similar compositions to neighboring, older biofilms. Apart from early changes in composition indicative of an initialization stage, biofilm communities in water storage facilities were temporally stable, although somewhat spatially heterogeneous. In contrast, suspended communities showed seasonal changes and were heavily influenced by water chemistry. Additionally, suspended communities were spatially homogeneous within a facility, and even at different facilities within the DWDS. In storage facilities that experienced problematic nitrification episodes and decreases in chloramine concentrations, suspended AOB concentrations increased as chloramine concentrations decreased. Notably, decreases in disinfectant were not accompanied by increases in the growth of other bacteria. Rather, as AOB concentrations increased, the total biomass of suspended communities actually decreased. During nitrification events, biofilm and suspended community compositions were most similar, lending further support to the concept that biofilms may act as reservoirs for nuisance and pathogenic bacteria in the DWDS. Abundant taxa were consistent with other studies of DWDSs that maintain chloramine, which provided support for the applicability of these findings to other systems, especially as there are no studies to compare to, to-date, of the microbiome in elevated storage towers.
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    Magnesium based materials and their antimicrobial activity
    (2011-09) Robinson, Duane Allan
    The overall goals of this body of work were to characterize the antimicrobial properties of magnesium (Mg) metal and nano-magnesium oxide (nMgO) in vitro, to evaluate the in vitro cytotoxicity of Mg metal, and to incorporate MgO nanoparticles into a polymeric implant coating and evaluate its in vitro antimicrobial properties. In the course of this work it was found that Mg metal, Mg-mesh, and nMgO have in vitro antimicrobial properties that are similar to a bactericidal antibiotic. For Mg metal, the mechanism of this activity appears to be related to an increase in pH (i.e. a more alkaline environment) and not an increase in Mg2+. Given that Mg-mesh is a Mg metal powder, the assumption is that it has the same mechanism of activity as Mg metal. The mechanism of activity for nMgO remains to be elucidated and may be related to a combination of interaction of the nanoparticles with the bacteria and the alkaline pH. It was further demonstrated that supernatants from suspensions of Mg-mesh and nMgO had the same antimicrobial effect as was noted when the particles were used. The supernatant from Mg-mesh and nMgO was also noted to prevent biofilm formation for two Staphylococcus strains. Finally, poly-ε-caprolactone (PCL) composites of Mg-mesh (PCL+Mg-mesh) and nMgO (PCL+nMgO) were produced. Coatings applied to screws inhibited growth of Escherichia coli and Pseudomonas aeruginosa and in thin disc format inhibited the growth of Staphylococcus aureus in addition to the E. coli and P. aeruginosa. Pure Mg metal was noted to have some cytotoxic effect on murine fibroblast and osteoblast cell lines, although this effect needs to be characterized further. To address the need for an in vivo model for evaluating implant associated infections, a new closed fracture osteomyelitis model in the femur of the rat was developed. Magnesium, a readily available and inexpensive metal was shown to have antimicrobial properties that appear to be related to its corrosion products and that nMgO has similar effects. Incorporation of nMgO into a PCL composite was easily achieved and revealed similar, although not identical antimicrobial results. This work has provided a strong foundation and methodology for further evaluation of Mg based materials and their antimicrobial properties.
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    Structure and activities of beta toxin: a virulence factor of Staphylococcus aureus
    (2009-06) Huseby, Medora Jean
    Beta toxin is a neutral sphingomyelinase secreted by certain strains of Staphylococcus aureus. This virulence factor lyses erythrocytes in order to evade the host immune system as well as to scavenge nutrients. The structure of beta toxin was determined at 2.4 Å resolution using crystals that were merohedrally twinned. This structure is similar to that of the endonuclease HAP1, Escherichia coli endonuclease III, bovine pancreatic DNase I, and the endonuclease domain of TRAS1 from Bombyx mori. Our biological assays demonstrated for the first time that beta toxin kills proliferating human lymphocytes. Structure-directed active site mutations show the biological activities of hemolysis and lymphotoxicity are due to the sphingomyelinase activity of the enzyme. The structures of all bacterial neutral sphingomyelinases solved to date reveal a solvent exposed hydrophobic beta hairpin. We examined the role of this beta hairpin in beta toxin virulence. Altering the length but not the content of the beta hairpin attenuates the biological activities associated with beta toxin. The beta hairpin is an important stabilizing structure. X-ray crystallographic analysis of beta hairpin mutants revealed very minimal structural changes. We show for the first time diacylglycerol bound in the beta toxin truncation (275-280) structure near the beta hairpin region. We also show at 1.75 Å resolution Mg2+ and phosphate bound to the F277A P289A structure. Neutral sphingomyelinases belong to the DNase I super-family of proteins (CATH class 3.60). Beta toxin shares the overall fold of DNase I with an RMSD value 3.3 Å over 220 Cαs. Beta toxin does not function as a DNase and instead precipitates nucleic acid. DNA causes beta toxin to non-specifically cross-link to other proteins. Extracellular DNA is a major structural component of the S. aureus biofilm matrix. Here we demonstrate that beta toxin has a profound effect on forming the matrix on which biofilms grow through the nucleic acid-dependent formation of cross-linked beta toxin monomers as well as other proteins. These links, plus the ability to bind eDNA, enables formation of the underlying nucleoprotein matrix essential to establish a biofilm. The goal of this thesis project is to understand the structural foundations for the role of the virulence factor beta toxin in order to understand the biological mechanism that allows S. aureus to successfully invade, colonize, and attack a host.
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    Time Dependent Effects of Contemporary Irrigants on a Polymicrobial Biofilm
    (2014-03) Barsness, Brian
    INTRODUCTION: Removal of contaminated canal contents and biofilms by mechanical preparation alone are limited. Irregular cross sections, lateral canals, and apical deltas are mostly inaccessible to mechanical preparation (Siqueira et al., 1997). Therefore, the use of chemical dissolution and disinfection of these regions is necessary. Many studies have demonstrated the efficacy of sodium hypochlorite in providing these functions (Hand et al., 1978; Rosenfeld et al., 1978). However, adverse cytotoxic outcomes have been reported with its use (Joffe et al., 1991), as well as reports of allergic hypersensitivity (Kaufman et al., 1989). The ideal irrigant has attributes of; broad spectrum antimicrobial activity, ability to dissolve pulp tissue remnants, inactivates endotoxin, removes the smear layer components, and is systemically nontoxic to periodontal tissues. Such an irrigant does not yet exist (Zehnder, 2006). MTAD, a final irrigation solution containing 3% doxycycline, 4.25% citric acid, and 0.5% Tween-80 has been shown to remove smear layer on extracted human teeth and provides broad spectrum antimicrobial activity (Torbinejad et al., 2003). QMiX, a final irrigation solution containing; bisbiguanide, calcium chelating agent, cetrimide surfactant, and saline has demonstrated efficacy at removing smear layer and disinfecting dentin tubules (Wang et al., 2013). SmearClear, a smear layer removing irrigant containing EDTA, cetrimide and water, has been shown to remove smear layer efficiently in an ex-vivo, split-tooth model (Andrabi et al., 2013). The aims of this study are to; demonstrate the use of a novel polymicrobial biofilm model to test endodontic irrigants, measure the susceptibility of a biofilm to individual irrigant treatment groups, and to characterize the responses visually through scanning electron microscopy (SEM) and laser confocal microscopy. METHODS: Clinical endodontic microbial samples were collected from six adult patients that presented to the graduate endodontics clinic at the University of Minnesota School of Dentistry with a diagnosis of pulpal necrosis. For each of the 3 experimental runs a pool of one to three patients were combined to ensure a collection of heterogeneous microbial species. A series of baffled 500 mL Erlenmeyer culture flasks were used as a “bioreactor” system to facilitate biofilm formation on hydroxyapatite discs. A total of six discs were contained per flask (2 discs per cell strainer basket). The flasks were covered with a sterile, vented flask cap and placed within the anaerobic chamber for incubation at 37°C and 20 RPM. This allowed for biofilm formation on the hydroxyapatite discs under shear force conditions. The flasks were allowed to incubate according to the conditions described above for approximately 48 total hours. Following incubation, discs were randomly selected for placement into a sterile, flat-bottomed, 12-well polystyrene culture plate (Sigma-Aldrich) for treatment according to the irrigant protocol.Four irrigation solutions were independently used for testing on the 48 hour biofilms. Sterile phosphate buffered saline (PBS) was used as a negative control treatment group. MTAD (DENTSPLY, Int.), QMiX (DENTSPLY, Int.), and Smearclear (SybronEndo, Orange, CA) were used as challenge treatment groups. Groups included; 1X PBS at 1 minute exposure, MTAD at 1 minute exposure,MTAD at 20 minutes exposure, QMiX at 1 minute exposure,QMiX at 20 minutes exposure,SmearClear at 1 minute exposure, and SmearClear at 20 minutes exposure. Evaluation of biofilm disruption was determined by; cell viability staining, crystal violet biomass staining, SEM, and confocal laser microscopy. Microbial speciation was performed by the Forsyth Institute (Cambridge, MA) by HOMIM 16sRNA analysis. RESULTS: The flask bioreactor, as previously described, can be utilized for growing a polymicrobial biofilm suitable for testing the efficacy of antimicrobial and biofilm removing endodontic irrigants. When compared as a group, the endodontic irrigants in this study achieved a statistically significant reduction in cell viability as time of exposure increased. In a pairwise comparison between irrigants, the mean cell viability was lowest for QMiX. No statistically significant difference in the reduction of biomass between endodontic irrigants was observed. Comparing endodontic irrigants individually by time did not result in a statistically significant difference in reducing cell viability or biomass. None of the endodontic irrigants completely removed the biofilm, as observed by SEM and confocal laser microscopy. 6 of 12 microbial species recovered were found to be in common between the patient pools when analyzed by HOMIM 16sRNA identification. This study presents a simple model for growing endodontic biofilms under anaerobic conditions. CONCLUSIONS: Findings from this study support the inefficient removal of biofilm by the irrigants included in the study. It was determined that QMiX achieved the lowest cell viability over the 20 minute treatment exposure. As a group, the irrigants were statistically significant in reducing cell viability and reducing biomass. However, it could be concluded that they should not be utilized as a single irrigant to disinfect or remove biofilm.
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    Time dependent effects of iodine potassium iodide (IKI) on a polymicrobial bioflm.
    (2012-08) Anders, Nicholas John
    Iodine potassium idodide is a less common endodontic irrigant that has been in use for years. Until now it has yet to be tested on a polymicrobial biofilm model. This study tested the feasibility of a new polymicrobial biofilm model for testing of endodontic irrigants by measuring remaining viability with MTT and SEM. There was no significant difference between time points between 1 and 20 minutes of 2% IKI application and 6% sodium hypochlorite. There was no significant difference between IKI at any time point and the PBS control for remaining bioburden. 6% sodium hypochlorite was significantly better at biofilm dissolution than IKI and PBS. 4. Due to the inability of 2 % IKI to remove biofilm it is not advised for use as the primary endodontic irrigant but may be a successful as an adjunctive irrigant to disinfect any residual biofilm that remains after cleaning and shaping. IKI may show potential as an intracanal medicament for species like E. faecalis that may be resistant to calcium hydroxide due to its powerful antimicrobial activity with the ability to penetrate biofilms.
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    Ultrastructural characterization of matrix development and the role of extracellular DNA in early Enterococcus faecalis biofilms
    (2012-11) Barnes, Aaron Michael Tolo
    Enterococcus faecalis is a highly adaptable, gram-positive bacterium that occupies a diverse range of ecological niches. A common soil-dwelling organism, it is also inhabits the metazoan gastrointestinal tract—from insects to humans. E. faecalis is remarkably resistant to a wide range of clinically-relevant antibiotics and readily forms biofilms on both abiotic and biotic surfaces. These latter factors underlie the medical relevance of E. faecalis. This thesis explores the ramifications of early developmental events in E. faecalis biofilm formation. Using correlative microscopy techniques, we investigated a series of mutants with ultrastructural changes in the extracellular matrix that elucidate the roles these genes play in matrix architecture. We also report that extracellular DNA plays a substantial role in stabilizing early (< 8 hr post-inoculation) E. faecalis biofilms, and that the source of this DNA is not via bulk cell lysis, but rather appears to be secreted from metabolically active cells. A putative model for this non-canonical source of DNA in the matrix is also proposed.
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    Zeolite Incorporated Materials for Targeted Biomass Retention and Pollutant Removal
    (2022-04) Chester, Anndee
    This dissertation describes the assessment and treatment of pollutants, namely nutrients, in waste streams. Nutrients such as nitrogen, are of major and growing concern because nitrogen removal from waste streams is energy and cost intensive; yet, without treatment cause eutrophication in aquatic systems. The aquatic health of the Volta River in Ghana was assessed by monitoring pollutants including water quality parameters, contaminants of emerging concern, antibiotic resistance, and the microbial community. While Ghana is a low- to middle- income country, inadequate sanitation infrastructure and environmental regulations contribute to environmental and human health issues. In this highly collaborative work, common (e.g., nitrogen) and emerging contaminants (e.g., DEET, PFAS) were detected and the microbial community was analyzed from samples collected along the length of the lower Volta River. Spikes in microbial detection (16S rRNA gene) and antibiotic resistant genes were associated with anthropogenic activities indicating adverse effects of human activities on the health of the Volta River. Additionally, novel biofilm technologies were explored to enhance nitrogen removal from waste streams. Specifically, zeolite-coated hollow fiber membranes and zeolite-coated biofilm carriers were designed to facilitate the partial nitritation-anammox (PNA) processes in mainstream wastewater, where significant cost savings and improved treatment could be realized. Zeolite particles and zeolite coated membranes in batch systems fed with mainstream-like synthetic wastewater demonstrated that anammox bacteria could be enriched and total nitrogen removal enhanced when compared to control systems without zeolite. By varying the mass of zeolite in the system it was discovered that a minimum amount of zeolite, or ammonium sorption capacity, was needed to achieve anammox retention. Zeolite-coated materials were further tested in flow-through systems to determine under what wastewater-relevant conditions nitrogen treatment enhanced. Zeolite-coated carriers in reactors under anaerobic conditions significantly retained anaerobic ammonia oxidizing (anammox) bacteria over systems with uncoated carriers; however, identical reactors operated under aerobic conditions did not retain aerobic oxidizing bacteria (AOB) on the carriers themselves. In both anaerobic and aerobic conditions, AOB were preferentially retained in the liquid of the reactors containing zeolite-coated carriers. Unexpectedly, denitrifying genes (specifically nirS, nirK, and nosZ) were also retained in systems with zeolite-coated carriers, indicating the nitrite-shunt process maybe another application. Zeolite-coated membranes were configured in flow-through membrane-aerated reactors and subject to varying operating lengths, inter-lumen oxygen concentrations, and influent nitrite with mixed results. Anammox bacteria were only detected in high quantities on zeolite membranes when operated for two weeks with 100% oxygen with and without nitrite in the influent. AOB were not enriched under any conditions at a 95% confident interval. Further exploration is needed to better understand the lack of AOB retention on both zeolite-carriers and membranes. Finally, zeolite-coated carriers were tested in stormwater-like systems both in the field and in laboratory reactors for retention of anammox, AOB, and feammox bacteria. Anammox bacteria and AOB were detected in increased quantities on zeolite-coated carriers over uncoated carriers when deployed in a raingarden, but not when deployed in a stormwater pond outlet structure. Carriers were also pre-seeded with anammox biofilm prior to field deployment in order to monitor biomass retention, and at the 2.5-month time scale tested, both control and zeolite carriers in both stormwater systems demonstrated excellent retention of biomass. Biomass was also well retained when both carrier types were pre-seeded and tested in laboratory reactors with simulated storm events. When pre-seeded, both reactors also demonstrated high rate of ammonium removal. Systems containing zeolite carriers inoculated with pond-water, however, had much higher rates of ammonium removal over control carriers indicating that under some conditions, zeolite coating did improve reactor performance. Finally, zeolite particles and zeolite-coated carriers were explored to determine if they also would preferentially retain feammox bacteria, the only known microorganism to defluorinate per- and polyfluorinated alkylated compounds. Reactors with zeolite particles and zeolite-coated carriers, had increased feammox bacteria and higher rates of ammonium removal. Overall, this research has demonstrated that zeolite-incorporated technologies are promising solutions to retaining anammox, AOB, and feammox bacteria and enhancing nitrogen removal in waste streams if applied under the right conditions. Treating waste streams to reduce the impacts of excess nutrients and other pollutants from human sources is important to protecting the health of aquatic systems.