Microbial fuel cells (MFCs) are electrochemical devices that exploit the ability of certain microorganisms to anaerobically respire using an insoluble terminal electron acceptor and therefore generate an electrical current. These bacteria are called electrogens or electrogenic bacteria. Two species of Arcobacter, Arcobacter butzleri ED-1 and Arcobacter L were isolated from the anodic chamber of an acetate fed MFC, and A. butzleri ED-1 was found to be the more electrogenic of the two bacteria. Arcobacter spp. are  proteobacteria and A. butzleri ED-1 and Arcobacter L were the first example of electrogenically active e proteobacteria. It was decided to study their interactions with the anode by fluorescent microscopy and study their electrogenic mechanisms by comparative proteomics using the iTRAQ method as it would allow for simultaneous identification and quantification of peptides in multiple samples. Fluorescent imaging over a period of 120 h in a half MFC showed that both A. butzleri ED-1 and Arcobacter L formed a thin anodic biofilm of a few cells thick and that A. butzleri ED-1 maintained a more stable anodic biofilm than Arcobacter L. iTRAQ analysis showed that the flagellin FlaA was up-regulated 2.4 fold at the anode but no other electron transport proteins or adhesins were upregulated. These results were distinct from those observed for other electrogenic bacteria (Geobacter sulfurreducens and Shewanella oneidensis MR-1) in previous studies which exhibited up-regulated electron transport proteins at the anode as well as forming an anodic biofilm of 50 μm thick. Therefore based on these results it was concluded that FlaA was most likely playing an important role A. butzleri ED-1 anode biofilm formation and that the mechanisms of electrogenesis in A. butzleri ED- 1 and Arcobacter L may be novel compared to those previously characterised. It was also concluded that one possible reason for A. butzleri ED-1 being more electrogenic than Arcobacter L was its ability to form a more stable anodic biofilm. It must be noted that both of these conclusions are highly speculative and further study is needed to elucidate the electrogenic mechanisms of A. butzleri ED-1 and to further compare biofilm formation between the two species.

With the current interest in alternative methods of energy production and increased utilization of existing energy sources, microbial fuel cells have become an important field of research. Microbial fuel cells are devices which harvest electrons from microorganisms created by their enzymatic oxidation of complex carbon substrates or consumed by their reduction of chemical oxidants. Microbial fuel cells with photosynthetic biocathodes are of particular interest due to their ability to simultaneously produce electricity and hydrocarbons while reducing carbon dioxide. Most species of microorganisms including many bacteria and yeasts require exogenous electron transfer mediators in order to allow electron transfer with an electrode. While adding such chemicals is simple enough at a lab scale, problems arise with chemical costs and separation at a larger scale. The goal of this research was to develop electrodes composed of a robust material which will eliminate the need for added soluble electron mediators in a photosynthetic biocathode microbial fuel cell. Electrodes made from stainless steel 304L have been coated in a conductive polymer (polypyrrole) and an immobilized electron transfer mediator (methylene blue) and tested chemically for stability and in a microbial fuel cell environment for use in bioanodes and biocathodes. The use of these immobilized mediator in the photosynthetic biocathode increased the open circuit voltage of the cell from 0.17 V to 0.24 V and the short circuit current from 8 mA/m2 to 64 mA/m2 (normalized to the geometric surface area of the electrode) when compared to using the same mediator in solution. The opposite effect was seen when using the electrodes in a bioanode utilizing Saccharomyces cerevisiae. The open circuit voltage decreased from 0.37 V to 0.31 V and the short circuit current decreased from 94 mA/m2 to 24 mA/m2 when comparing the immobilized mediator to soluble mediators. The impact of the membrane and pH of the anode and cathode solutions were quantified and were found to have much less of an effect on the internal resistance than the microbial factors.

With the current interest in alternative methods of energy production and increased utilization of existing energy sources, microbial fuel cells have become an important field of research. Microbial fuel cells are devices which harvest electrons from microorganisms created by their enzymatic oxidation of complex carbon substrates or consumed by their reduction of chemical oxidants. Microbial fuel cells with photosynthetic biocathodes are of particular interest due to their ability to simultaneously produce electricity and hydrocarbons while reducing carbon dioxide. Most species of microorganisms including many bacteria and yeasts require exogenous electron transfer mediators in order to allow electron transfer with an electrode. While adding such chemicals is simple enough at a lab scale, problems arise with chemical costs and separation at a larger scale. The goal of this research was to develop electrodes composed of a robust material which will eliminate the need for added soluble electron mediators in a photosynthetic biocathode microbial fuel cell. Electrodes made from stainless steel 304L have been coated in a conductive polymer (polypyrrole) and an immobilized electron transfer mediator (methylene blue) and tested chemically for stability and in a microbial fuel cell environment for use in bioanodes and biocathodes. The use of these immobilized mediator in the photosynthetic biocathode increased the open circuit voltage of the cell from 0.17 V to 0.24 V and the short circuit current from 8 mA/m2 to 64 mA/m2 (normalized to the geometric surface area of the electrode) when compared to using the same mediator in solution. The opposite effect was seen when using the electrodes in a bioanode utilizing Saccharomyces cerevisiae. The open circuit voltage decreased from 0.37 V to 0.31 V and the short circuit current decreased from 94 mA/m2 to 24 mA/m2 when comparing the immobilized mediator to soluble mediators. The impact of the membrane and pH of the anode and cathode solutions were quantified and were found to have much less of an effect on the internal resistance than the microbial factors.

Many of the most pressing global challenges today and in the future center around the scarcity of sustainable energy and water sources. The innovative microbial fuel cell (MFC) technology addresses both as it utilizes bacteria to convert wastewaters into electricity. Advancing this technology requires a better understanding of the optimal materials, designs and conditions involved. The micro-sized MFC was recently developed to serve this need by providing a rapid testing device requiring only a fraction of the materials. Further, development of micro-liter scale MFCs has expanded into potential applications such as remote and self-sustained power sources as well as on-chip energy generators. By using microfabrication, the fabrication and assembly of microsized MFCs is potentially inexpensive and mass produced. The objective of the work within this dissertation was to explore and optimize the micro-sized MFC to maximize power and current generation towards the goal of a usable and application-oriented device. Micro-sized MFCs were examined and developed using four parameters/themes considered most important in producing a high power generating, yet usable device: Anode- The use of nano-engineered carbon nanomaterials, carbon nanotubes and graphene, as anode as well as testing semiconductor industry standard anode contact area materials for enhanced current production. 5 Cathode- The introduction of a membrane-less air cathode to eliminate the need for continuous chemical refills and making the entire device mobile. Reactor design- The testing of four different reactor designs (1-75 μLs) with various features intended to increase sustainability, cost-effectiveness, and usability of the microsized MFC. Fuels- The utilization of real-world fuels, such as industrial wastewaters and saliva, to power micro-sized MFCs. The micro-sized MFC can be tailored to fit a variety of applications by varying these parameters. The device with the highest power production here was designed to be an inexpensive and robust power source in applications like point-of-care diagnostics in developing countries. This 25 μL graphene nanomaterial anode, air cathode device in an inexpensive flexible rubber architecture was powered by saliva and achieved 3.55 μW/cm2 and 35.2 W/m3. The continued optimization of MFC technology promises many interesting and innovative applications.

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Microbial fuel cells (MiFCs) have been suggested as a means to harness energy that is otherwise unutilized during the wastewater treatment process. MiFCs have the unique ability to treat influent waste streams while simultaneously generating power which can offset energy associated with the biological treatment of wastewater. During the oxidation of organic and inorganic wastes, microorganisms known as exoelectrogens have the ability to move electrons extracellularly. MiFCs generate electricity by facilitating the microbial transfer of these electrons from soluble electron donors in feedstocks to a solid-state anode. While MiFCs are a promising renewable energy technology, current systems suffer from low power densities which hinder their practical applicability. In this study, a novel anode design using flame-deposited carbon nanostructures (CNSs) on stainless steel mesh is developed to improve the electron transfer efficiency of electrons from microorganisms to the anode and thus the power densities achievable by MiFCs. These new anodes appear to allow for increased biomass accumulation on the anode and may aid in the direct transfer of electrons to the anode in mediatorless MiFC systems. Experiments were conducted using anaerobic biomass in single-chamber MiFCs with CNS-enhanced and untreated stainless steel anodes. Fuel cells utilizing CNS-enhanced anodes generated currents up to two orders of magnitude greater than cells with untreated metal anodes, with the highest power density achieved being 510 mW m-2. / Master of Science

Effective wastewater treatment is needed to reduce the water pollution problem. However, massive energy is consumed in wastewater treatment, required to design an innovative system to reduce the energy consumption to solve the energy crisis. Integrated photobioelectrochemical system (IPB) is a powerful system to combine microbial fuel cells (MFCs) and algal bioreactor together. This system has good performance on the organic degradation, removal of nitrogen and phosphorus, and recover the bioenergy via electricity generation and algal harvesting. This dissertation is divided to twelve chapters, about various aspects of the working mechanisms and actual application of IPB. Chapter 1 generally introduces the working mechanisms of MFCs, algal bioreactor, and modeling. Chapter 2 demonstrates the improvement of cathode material to improve the structure and catalytic performance to improve the MFC performance. Chapter 3 describes the process to use microbial electrolysis cell (MEC) to generate biohythane for the energy recovery. Chapters 4 and 5 demonstrate the application of stable isotope probing to study Shewanella oneidensis MR-1 in the MFCs. Chapters 6 to 8 describe the application of models to optimize MFC and IPB system performance. Chapter 9 describes the strategy improvement for the algal harvesting in IPB. Chapter 10 describes the application of scale-up bioelectrochemical systems on the long-term wastewater treatment. Chapter 11 finally concludes the perspectives of IPBs in the wastewater treatment and energy recovery. This dissertation comprehensively introduces IPB systems in the energy recovery and sustainable wastewater treatment in the future. / Ph. D. / The resource of pure water becomes more and more valuable, and the large discharge of the wastewater into the environment would even cause the environmental pollution. Thus, the wastewater is a necessary method to remove the organics out of the wastewater. However, the large energy consumption is a critical issue to solve due to the global energy burden. How to reduce the energy consumption in the wastewater treatment is the required step to achieve the sustainable water treatment. Integrated photobioelectrochemical system (IPB) is a new promising technology, alternative to the traditional wastewater treatment techniques (e.g., anaerobic digester or activated sludge reactor) with low energy consumption. The IPB system was to combine microbial fuel cells (MFCs), which is a typical bioelectrochemical system (BES), and the algal bioreactor together, to achieve the performance on the organic degradation, removal of nitrogen and phosphorus in the wastewater, and recover the bioenergy via electricity generation and algal harvesting. The system was proved to be effective, but most of the IPB systems were only proved to work in the laboratories, and there is still a large potential space to improve the IPB system performance in the actual environment. Herein, this dissertation combines multiple studies about the IPB improvement and scaled-up process in the real wastewater treatment. Chapter 1 generally introduces what are MFCs, algal bioreactor and modeling simulations. Chapter 2 demonstrates the method about how to improve the MFC material to enhance the treatment performance for better MFC performance. Chapter 3 describes how to use BES to convert the organics to the renewable gas (e.g., H₂ and CH₄) to recover the energy. Chapters 4 and 5 demonstrate the application of stable isotope probing to study the microbial behavior in the MFC. Chapters 6 to 8 describe the applications of model simulations to optimize MFC and IPB performance. Chapter 9 describes the new reactor to improve the algal harvesting process to obtain more energy from the IPB system. Chapter 10 describes how to use the scale-up IPB system to treat the real wastewater treatment. Chapter 11 finally puts forward some perspectives of IPBs in the wastewater treatment and energy recovery. This dissertation comprehensively gives a big picture about the development of IPB systems in the energy recovery and sustainable wastewater treatment in the future.

Bacteria in nature predominantly grow as biofilms on living and non-living surfaces. The development of biofilms on non-living surfaces is significantly affected by the surface micro/nano topography. The main goal of this dissertation is to study the interaction between microorganisms and nanopatterned surfaces. In order to engineer the surface with well-defined and repeatable nanoscale structures, a new, versatile and scalable nanofabrication method, termed Spun-Wrapped Aligned Nanofiber lithography (SWAN lithography) was developed. This technique enables high throughput fabrication of micro/nano-scale structures on planar and highly non-planar 3D objects with lateral feature size ranging from sub-50 nm to a few microns, which is difficult to achieve by any other method at present. This nanolithography technique was then utilized to fabricate nanostructured electrode surfaces to investigate the role of surface nanostructure size (i.e. 115 nm and 300 nm high) in current production of microbial fuel cells (MFCs). Through comparing the S. oneidensis attachment density and current density (normalized by surface area), we demonstrated the effect of the surface feature size which is independent of the effect on the surface area. In order to better understand the mechanism of microorganism adhesion on nanostructured surfaces, we developed a biophysical model that calculates the total energy of adhered cells as a function of nanostructure size and spacing. Using this model, we predict the attachment density trend for Candida albicans on nanofiber-textured surfaces. The model can be applied at the population level to design surface nanostructures that reduce cell attachment on medical catheters. The biophysical model was also utilized to study the motion of a single Candida albicans yeast cell and to identify the optimal attachment location on nanofiber coated surfaces, thus leading to a better understanding of the cell-substrate interaction upon attachment. / Ph. D. / Formation of surface associated multicellular communities of microorganisms known as biofilms is of concern in medical settings as well as in industries such as oil refineries and marine engineering. It has been shown that micro/nanoscale surface features can highly regulate the process of biofilm formation and the attached cell activities. In this dissertation, we study the interaction between surface nanoscale structures and bacterial adhesion by experiments and biophysical modelling. We develop the Spun-Wrapped Aligned Nanofiber (SWAN) lithography, a versatile, scalable, and high throughput technique for masterless nanopatterning of hard materials. Using this technique, we demonstrate high fidelity whole surface single step nanopatterning of bulk and thin film surfaces of regularly and irregularly shaped 3D objects. SWAN lithography is used to texturize the electrode surface of microbial fuel cells (MFCs), which are envisioned as an alternative sustainable energy source. Compared to the non-patterned electrodes, the electrodes with 115 nm surface patterns facilitate larger biofilm coverage and 40% higher current production. We also develop a biophysical model to optimally texturize the surface of central venous and uretic medical catheters to prevent biofilm formation by fungal pathogen, Candida albicans. We show that the surface structures that result the highest cell total energy retained the least C. albicans. Furthermore, the adhesion behaviour of a single yeast cell is also experimentally studied in conjunction with the developed model.