In order to optimize the ability of Geobacter sulfurreducens to produce electrical current and remediate wastewater, several physiological challenges must be overcome. The accumulation of protons at the electrode surface of a microbial fuel cell (MFC) decreases the pH, and,…
In order to optimize the ability of Geobacter sulfurreducens to produce electrical current and remediate wastewater, several physiological challenges must be overcome. The accumulation of protons at the electrode surface of a microbial fuel cell (MFC) decreases the pH, and, thus, the ability of the bacteria to maintain baseline metabolic conditions. To evaluate the extent to which this pH change hinders performance, the buffer concentration supplied to G. sulfurreducens reactors was varied. The resulting biofilms were subjected to chronoamperometry, cyclic voltammetry, and confocal microscopy to determine metabolic function and biofilm thickness. Biofilms grown with a 30-mM bicarbonate buffer experienced limitations on cell function and current output due to proton accumulation, while 90- and 150-mM conditions alleviated these limitations most of the measurements. Based on the current output, estimated biofilm thickness, and the medium-rate and slow-rate scan rate cyclic voltammetry, benefits exist for buffer concentrations greater than 30 mM. If the kinetics of G. sulfurreducens electron transfer are optimized, the potential of the technique to be implemented for energy recovery is improved.
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Mining-influenced water (MIW) is an acidic stream containing a typically acidic pH (e.g., 2.5), sulfate, and dissolved metal(loid)s. MIW has the potential to affect freshwater ecosystems and thus MIW requires strategies put in place for containment and treatment. …
Mining-influenced water (MIW) is an acidic stream containing a typically acidic pH (e.g., 2.5), sulfate, and dissolved metal(loid)s. MIW has the potential to affect freshwater ecosystems and thus MIW requires strategies put in place for containment and treatment. Lignocellulosic sulfate-reducing biochemical reactors (SRBRs) are considered a cost-effective passive treatment for MIW and have been documented to continuously treat MIW at the field-scale. However, long-term operation (> 1 year) and reliable MIW treatment by SRBRs at mining sites is challenged by the decline in sulfate-reduction, the key treatment mechanism for metal(loid) immobilization. This dissertation addresses operational designs and materials suited to promote sulfate reduction in lignocellulosic SRBRs treating MIW. In this dissertation I demonstrated that lignocellulosic SRBRs containing spent brewing grains and/or sugarcane bagasse can be acclimated in continuous mode at hydraulic retention times (HRTs) of 7-12 d while simultaneously removing 80 ± 20% – 91 ± 3% sulfate and > 98% metal(loid)s. Additionally, I showed that decreasing the HRT to 3 d further yields high metal(loid) removal (97.5 ± 1.3% – 98.8 ± 0.9%). Next, I verified the utility of basic oxygen furnace slag to increase MIW pH in a two-stage treatment involving a slag stage and an SRBR stage containing spent brewing grains or sugarcane bagasse. The slag reactor from the two-stage treatment increased MIW pH from 2.6 ± 0.2 to 12 ± 0.3 requiring its re-combination with fresh MIW to reduce pH to 5.0 ± 1.0 prior to entering the lignocellulosic SRBRs. The lignocellulosic SRBRs from the two-stage treatment successfully continued to remove metal(loid)s, most notably cadmium, copper, and zinc at ≥ 96%. In additions to these outcomes, I performed a metadata analysis of 27 SRBRs employing brewers spent grains, sugarcane bagasse, rice husks and rice bran, or a mixture of walnut shells, woodchips, and alfalfa. I found that sugarcane bagasse SRBRs can remove between 94 and 168 mg metal(loid) kg–1 lignocellulose d–1. In addition, Bacteroidia relative abundances showed a positive correlation with increasing sulfate removal across all 27 SRBRs and are likely essential for the degradation of lignocellulose providing electron donors for sulfate reduction. Clostridia and Gammaproteobacteria were negatively correlated with sulfate reduction in the 27 SRBRs, however SRBRs that received alkalinized MIW had lower relative abundances of Clostridia, Gammaproteobacteria, and methanogenic archaea (known competitors for sulfate-reducing bacteria). Overall, my dissertation provides insight into lignocellulosic materials and operational designs to promote long-term sulfate-reduction in lignocellulosic SRBRs treating MIW.
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Bacterial biofilms exist on surfaces within pressurized water systems, posing threats to water quality and causing fouling or microbial induced corrosion. Germicidal UV irradiation has shown promise in deactivating planktonic pathogens in water but challenges in delivering light to surfaces…
Bacterial biofilms exist on surfaces within pressurized water systems, posing threats to water quality and causing fouling or microbial induced corrosion. Germicidal UV irradiation has shown promise in deactivating planktonic pathogens in water but challenges in delivering light to surfaces where biofilms exist have limited advancement in understanding biofilm response to UV-C light. This dissertation aims to overcome the limitation of delivering UV-C light through use of side-emitting optical fibers (SEOFs), advance capabilities to produce SEOFs and understand if a minimum UV-C irradiance can prevent biofilm formation. Two scalable manufacturing approaches were developed for producing kilometer lengths of thin (≤500-µm) and physically flexible SEOFs. One strategy involved dip-coating amine-functionalized SiO2 nanoparticles (NPs) on bare optical fiber, followed by a coating of UV-C transparent polymer (CyTop). I showed that NPs closer to the surface achieved with higher ionic strength solutions increased side-scattering of UV-C light. This phenomenon was primarily attributed to the interaction between NPs and evanescent wave energy. The second strategy omitted NPs but utilized a post-treatment to the UV-C transparent polymer that increased surface roughness on the outer fiber surface. This modification maintained the physical flexibility of the fiber while promoting side-emission of UV-C light. The side emission was due to the enhancement of refracted light energy. Both methods were successfully scaled up for potential commercial production.
Experimental platforms were created to study biofilm responses to UV light on metal or flexible plastic pipe (1/4” ID) surfaces. Delivering UV-C light via SEOFs with irradiances >8 µW/cm2 inhibited biofilm accumulation. Neither UV-A nor UV-B light inhibited biofilm growth. At very low UV-C irradiance (<3 µW/cm2), biofilms were not inhibited. Functional genomic analysis revealed that biofilms irradiated by insufficient UV-C irradiance upregulated various essential genes related to DNA repair, energy metabolism, quorum sensing, mobility, and EPS synthesis. When net UV-C biofilm inactivation rates exceeded the biofilm growth rate, biofilms were inhibited. Insights gained from this dissertation work shed light on the prospective applications of UV-C technology in addressing biofilm challenges within water infrastructure across multiple sectors, from potable water to healthcare applications.
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Due to the use of fertilizers, concentrations of harmful nitrate have increased in groundwater and surface waters globally in the last century. Water treatment plants primarily use separation techniques for nitrate treatment, but these technologies create a high nitrate concentration…
Due to the use of fertilizers, concentrations of harmful nitrate have increased in groundwater and surface waters globally in the last century. Water treatment plants primarily use separation techniques for nitrate treatment, but these technologies create a high nitrate concentration brine that is costly to dispose of. This dissertation focuses on catalytic hydrogenation, an emerging technology capable of reducing nitrate to nitrogen gas using hydrogen gas (H2). This technology reduces nitrate at rates >95% and is an improvement over technologies used at water treatment plants, because the nitrate is chemically transformed with harmless byproducts and no nitrate brine. The goal of this dissertation is to upgrade the maturity of catalytic nitrate hydrogenation systems by overcoming several barriers hindering the scale-up of this technology. Objective 1 is to compare different methods of attaching the bimetallic catalyst to a hollow-fiber membrane surface to find a method that results in 1) minimized catalyst loss, and 2) repeatable nitrate removal over several cycles. Results showed that the In-Situ MCfR-H2 deposition was successful in reducing nitrate at a rate of 1.1 min-1gPd-1 and lost less than 0.05% of attached Pd and In cumulatively over three nitrate treatment cycles. Objective 2 is to synthesize catalyst-films with varied In3+ precursor decorated over a Pd0 surface to show the technology can 1) reliably synthesize In-Pd catalyst-films with varied bimetallic ratios, and 2) optimize nitrate removal activity by varying In-Pd ratio. Results showed that nitrate removal activity was optimized with a rate constant of 0.190 mg*min-1L-1 using a catalyst-film with a 0.045 In-Pd ratio. Objective 3 is to perform nitrate reduction in a continuous flow reactor for two months to determine if nitrate removal activity can be sustained over extended operation and identify methods to overcome catalyst deactivation. Results showed that a combination of increased hydraulic residence time and reduced pH was successful in increasing the nitrate removal and decreasing harmful nitrite byproduct selectivity to 0%. These objectives increased the technology readiness of this technology by enabling the reuse of the catalyst, maximizing nitrate reduction activity, and achieving long-term nitrate removal.
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While most household surfactants are biodegradable in aerobic conditions, their presence in a microbiological treatment process can lead to the proliferation of antimicrobial-resistance genes (ARG) in bacteria, such as Pseudomonas aeruginosa. Surfactants can be cationic, anionic, or zwitterionic, and…
While most household surfactants are biodegradable in aerobic conditions, their presence in a microbiological treatment process can lead to the proliferation of antimicrobial-resistance genes (ARG) in bacteria, such as Pseudomonas aeruginosa. Surfactants can be cationic, anionic, or zwitterionic, and these different classes may have different effects on the proliferation of ARG. This study evaluated how the three classes of surfactants affected the microbial community’s structure and ARG in O2-based membrane biofilm reactors (O2-MBfRs) that provided at least 98% surfactant removal. Cationic cetrimonium bromide (CTAB) had by far the strongest impact with highest ARG abundance in the biofilm. In particular, Pseudomonas and Stenotrophomonas, the two main genera in the biofilm treating CTAB, were highly correlated to the abundance of ARG for efflux pumps and antibiotic inactivation. CTAB also promoted potential of horizontal gene transfer (HGT) of ARG. Combining results from the metabolome and metagenome identified four possible pathways for CTAB biodegradation. Of special important is a new pathway: β-carbon oxidation of CTAB to produce betaine. An insufficient nitrogen source could lead to irreversible ARB and ARG enrichment in the MBfR biofilm. Finally, a two-stage O2-MBfR successfully removed a high concentration (730 mg/L) of CTAB: Partial CTAB removal in the Lead reactor relieved inhibition in the Lag reactor. Metagenomic analysis also revealed that the Lag reactor was enriched in genes for CTAB and metabolite oxygenation.
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Cyanobacteria and microalgae help reduce the environmental impact of human energy consumption by playing a vital role in carbon and nitrogen cycling. They are also used in various applications like biofuel production, food, medicine, and bioremediation. Understanding how these organisms…
Cyanobacteria and microalgae help reduce the environmental impact of human energy consumption by playing a vital role in carbon and nitrogen cycling. They are also used in various applications like biofuel production, food, medicine, and bioremediation. Understanding how these organisms respond to stress is important for efficient recovery strategies and sustainable outcomes. This study investigated the effects of low-level bleaching and thermal stress on cyanobacteria and microalgae, specifically Synechocystis, Chlorella, and Scenedesmus. The role of ferroptosis, an iron-dependent form of cell death, in the degradation of cellular components under these stressors was examined. Flow cytometry and spectrophotometry were used to measure changes in cellular health and viability. The results showed that temperature influences the type of cell death mechanism and can impact photosynthetic organisms. When treated with Liproxstatin-1, an inhibitor of ferroptosis, both Synechocystis and Chlorella experienced a decrease in oxidative damage, suggesting a potential protective role for the compound. Further investigation into ferroptosis and other forms of cell death, as well as identifying additional inhibitory molecules, could lead to strategies for mitigating oxidative stress and enhancing the resilience of cyanobacteria and microalgae.
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Methane (CH4) is a prominent greenhouse gas that contributes to the negative impacts of global warming and climate change, whose emissions have more than doubled since the Industrial Revolution primarily due to anthropogenic sources. The main pathways in which methane…
Methane (CH4) is a prominent greenhouse gas that contributes to the negative impacts of global warming and climate change, whose emissions have more than doubled since the Industrial Revolution primarily due to anthropogenic sources. The main pathways in which methane moves through the environment are methanogenesis and methanotrophy. Methane is primarily generated by acetoclastic methanogenesis in wetlands while it can be oxidized both aerobically and anaerobically. Wetlands are important methane emission sources at 177 - 284 Tg CH4 year-1. The Tres Rios Wetland (TRW) is a constructed facility to complete nutrient removal of treated municipal wastewater, and has shown low emissions of methane. Whether such low emissions could be achieved through active anaerobic oxidation of methane (AOM) is not known, and the main objective of this work is to evaluate the rates of AOM in TRW. In this study an isotopic method and a mass balance method were utilized to determine the rate of AOM from top sediments found at Tres Rios at various locations and in two sets of sampling. The results showed that evidence of AOM occurred in the sediments of both sampling events conducted. The first sampling set showed evidence of AOM at all locations along a transect, showing that oxidation of methane is indeed occurring in Tres Rios sediments. Evidence from both methodologies suggested that high methanogenesis rates occurred at the outside location closest to the water. The second sampling set showed that the highest rate of AOM occurred at the outlet location, with the lowest rate occurring in the middle location. DNA extractions and PCR images resulted in a poor DNA yield, and inability to extract DNA. It was determined that the isotopic approach was less accurate than the mass balance approach due to unexpected delta CH4 values. It was determined that dilutions of CH4 ppm lead to less accurate isotopic measurements needed to estimate AOM rates using a 13C pulse technique. Literature review suggests that factors including water presence, temperature, redox potential, and plant presence can be influential in the oxidation of methane. This AOM assay can be beneficial in better understanding how methane cycles at Tres Rios, and can provide opportunities for future research in determining which factors influence the oxidation of methane in different locations throughout wetlands.
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Widespread use of halogenated organic compounds for commercial and industrial purposes makes halogenated organic pollutants (HOPs) a global challenge for environmental quality. Current wastewater treatment plants (WWTPs) are successful at reducing chemical oxygen demand (COD), but the removal of HOPs…
Widespread use of halogenated organic compounds for commercial and industrial purposes makes halogenated organic pollutants (HOPs) a global challenge for environmental quality. Current wastewater treatment plants (WWTPs) are successful at reducing chemical oxygen demand (COD), but the removal of HOPs often is poor. Since HOPs are xenobiotics, the biodegradation of HOPs is usually limited in the WWTPs. The current methods for HOPs treatments (e.g., chemical, photochemical, electrochemical, and biological methods) do have their limitations for practical applications. Therefore, a combination of catalytic and biological treatment methods may overcome the challenges of HOPs removal.This dissertation investigated a novel catalytic and biological synergistic platform to treat HOPs. 4-chlorophenol (4-CP) and halogenated herbicides were used as model pollutants for the HOPs removal tests. The biological part of experiments documented successful co-oxidation of HOPs and analog non-halogenated organic pollutants (OPs) (as the primary substrates) in the continuous operation of O2-based membrane biofilm reactor (O2-MBfR). In the first stage of the synergistic platform, HOPs were reductively dehalogenated to less toxic and more biodegradable OPs during continuous operation of a H2-based membrane catalytic-film reactor (H2-MCfR). The synergistic platform experiments demonstrated that OPs generated in the H2-MCfR were used as the primary substrates to support the co-oxidation of HOPs in the subsequent O2-MBfR. Once at least 90% conversation of HOPs to OPs was achieved in the H2-MCfR, the products (OPs to HOPs mole ratio >9) in the effluent could be completely mineralized through co-oxidation in O2-MBfR. By using H2 gas as the primary substrate, instead adding the analog OP, the synergistic platform greatly reduced chemical costs and carbon-dioxide emissions during HOPs co-oxidation.
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The waterways in the United States are polluted by agricultural, mining, and industrial activities. Recovery of valuable materials, such as energy and nutrients, from these waste streams can improve the economic and environmental sustainability of wastewater treatment. A number of…
The waterways in the United States are polluted by agricultural, mining, and industrial activities. Recovery of valuable materials, such as energy and nutrients, from these waste streams can improve the economic and environmental sustainability of wastewater treatment. A number of state-of-the-art anaerobic bioreactors have promise for intensified anaerobic biological treatment and energy recovery, but they have drawbacks. The drawbacks should be overcome with a novel anaerobic biological wastewater treatment process: the anaerobic biofilm membrane bioreactor (AnBfMBR). This research works aims to advance key components of the AnBfMBR. The AnBfMBR is a hybrid suspended growth and biofilm reactor. The two main components of an AnBfMBR are plastic biofilm carriers and membranes. The plastic biofilm carriers provide the surface onto which the biofilms grow. Membranes provide liquid-solid separation, retention of suspended biomass, and a solids-free effluent. Introducing sufficient surface area promotes the biofilm accumulation of slow-growing methanogens that convert volatile fatty acids into methane gas. Biofilms growing on these surfaces will have a mixed culture that primarily consists of methanogens and inert particulate solids, but also includes some acetogens. Biomass that detaches from biofilms become a component of the suspended growth. A bench-scale AnBfMBR was designed by the AnBfMBR project team and constructed by SafBon Water Technology (SWT). The primary objective of this thesis project was to evaluate the ability of plastic biofilm carriers to minimize ceramic-membrane fouling in the AnBfMBR setting. A systematic analysis of mixing for the bench-scale AnBfMBR was also conducted with the plastic biofilm carriers. Experiments were conducted following a ‘run to failure’ method, in which the ceramic membranes provide filtration, and the time it takes to reach a ‘failure transmembrane pressure (TMP)’ was recorded. The experiments revealed two distinct trends. First, the time to failure TMP decreased as mixed liquor suspended solids concentration (MLSS) concentration increased. Second, increasing the carrier fill extend the time to failure, particularly for higher MLSS concentrations. Taken together, the experiments identified an optimized “sweet spot” for the AnBfMBR: an operating flux of 0.25-m/d, a failure TMP of 0.3-atm pressure, MLSS of 5,000 – 7,500 mg/L, and 40% carrier fill.
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Anaerobic Digestion (AD) typically stabilizes 40-60% of influent wastewater sludge. Improving the methane yield in wastewater may produce enough energy to power some wastewater treatment processes, while the production of volatile-fatty acids (VFAs) generates economic incentives for yard waste pre-fermentation.…
Anaerobic Digestion (AD) typically stabilizes 40-60% of influent wastewater sludge. Improving the methane yield in wastewater may produce enough energy to power some wastewater treatment processes, while the production of volatile-fatty acids (VFAs) generates economic incentives for yard waste pre-fermentation. In this research, pre-fermenters consisting of inocula composed of media; cellulose, lantana, or grass; and rabbit cecotrope were fed various concentrations of plant matter. The contents of these pre-fermenters were the influent for respective anaerobic digesters. The microbial consortium derived for the lignocellulosic pretreatment with common yard waste in Arizona successfully increased methane production in AD, while producing additional VFAs during pretreatment in all systems. The performance of the system appeared to depend on plant matter loading and operating time, with a higher plant loading increasing the VFA production and a longer operating time increasing soluble chemical oxygen demand (COD) in pre-fermentation, and therefore the methane production in AD increased. The pre-fermenter with the highest plant matter loading and longest operating time –1.44 g plant matter per day at a 9.6% influent concentration and 193 days of total operating time– produced 10,000 mg COD/L of VFA, and its reactor produced about 460 mL methane (CH4) per day, which was almost twice the production of the control AD at 250 mL CH4 per day. This research uses yard waste that would previously be disposed of in landfill to increase valuable product production in AD. The potential value added to wastewater treatment plant (WWTP) processes by these methods could incentivize the expansion of wastewater treatment, thereby increasing sanitation access. The use of net-neutral biogas as a fuel source for WWTPs is additionally an incremental solution for reducing carbon equivalents present in the atmosphere, thereby reducing the greenhouse gas effect.
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