Submitted by Marilene Donadel (marilene.donadel@unioeste.br) on 2017-08-18T21:49:26Z No. of bitstreams: 1 Simone C Rodrigues 2016.pdf: 1227135 bytes, checksum: 39f934de9e9e731081567168e81f33ef (MD5) / Made available in DSpace on 2017-08-18T21:49:26Z (GMT). No. of bitstreams: 1 Simone C Rodrigues 2016.pdf: 1227135 bytes, checksum: 39f934de9e9e731081567168e81f33ef (MD5) Previous issue date: 2016-09-09 / For some years, the environment has been one of the main concerns of the global population. The need for alternative sources and ecologically friendly practices, as well as biofuels, is an option that fits sustainable development. In search for high oil content raw materials, microalgae have been considered as a useful and promising source of biodiesel. Thus, the paper aimed to evaluate the production of ethyl esters via in situ reaction of the biomass obtained in the cultivation Chlamydomonas reinhardtii microalgae, to determine the best cultivation medium based on productivity, to develop microalgae cultivation for biomass production, to determine lipid content, the in situ reaction for ethyl esters production characterize and quantify the obtained esters. From the results obtained with the extraction, it was observed that the characteristics of the solvents can promote the lipid extraction of the biomass. Chlamydomonas reinhardtii microalgae was cultivated in TAP medium, with controlled artificial light and temperature. Then, there was the recovery of the biomass, which analyzed for chlorophyll a and b proteins and total carbohydrates and finally, the lipid content in the biomass was obtained and quantified by analyzing the ethyl esters present by gas chromatography. The chlorophyll, protein and carbohydrate contents were: 54.136 mg / L -1, 29.39% and 14.92%, respectively, and the highest lipid content was 4.03%. The yield of the biomass culture was of 0.646 gL-1. From the results obtained with the extraction, it was observed that, the polarity of the solvent influences the productivity of lipids. In the production of esters the predominant in situ reaction is the temperature and reaction time, with the highest yields at 60ºC (1.504 mg of ester/g of biomass in 30, 1.518 mg of ester/g of biomass in 45 and 1.509 mg of ester/g of biomass in 60 minutes). In the production of esters, the factor analysis showed that there was interaction between temperature and time, on average, the best yields in number of C15 and C16 cetanes being C16 (76.97% at 30°C, 58.21% at 45°C, 64.05 % In 60°C and 56.79% in 75°C) profile of esters obtained, suitable for the production of biodiesel. The obtained results showed that the Chlamydomonas reinhardtii microalgae biomass presents potential for biodiesel production, when cultivated under the conditions tested in this study. / O meio ambiente há alguns anos vem sendo uma das principais preocupações da população mundial. A necessidade de fontes alternativas de energia e de práticas ecologicamente corretas, assim como os biocombustíveis, surgem como uma opção que se ajusta ao desenvolvimento sustentável. Na busca por matérias primas com alto teor de óleo, as microalgas têm sido consideradas como uma fonte útil e promissora de biodiesel. Desta forma, o trabalho teve como objetivo a avaliação da produção de ésteres etílicos via reação in situ da biomassa obtida no cultivo da microalga Chlamydomonas reinhardtii, determinar o melhor meio de cultivo baseado na produtividade, desenvolver o cultivo de microalga para a produção de biomassa, determinar o teor de lipídios, a reação in situ para a produção de ésteres etílicos caracterizar e quantificar os ésteres obtidos. A partir dos resultados obtidos com a extração, observou-se que as características dos solventes podem favorecer a extração lipídica da biomassa. A microalga Chlamydomonas reinhardtii foi cultivada em meio TAP, com luz artificial e temperatura contraladas. Após houve a recuperação da biomassa, esta foi analisada quanto ao teor de clorofila a e b proteínas e carboidratos totais e por fim, foi obtido e quantificado o teor lipídico presente na biomassa, analisando-se o perfil de ésteres etílicos presentes por cromatografia gasosa. O teor de clorofila, proteína e carboidratos encontrados foram: 54,136 mg./L-1, 29,39% e 14,92%, respectivamente, e o maior teor de lipídeo foi de 4,03% o rendimento do cultivo de biomassa foi de 0,646 g.L-1. A partir dos resultados obtidos com a extração, observou-se que, a polaridade do solvente influencia na produtividade de lipídios. Na produção de ésteres o que predomina na reação in situ é a temperatura e o tempo de reação, apresentando maiores rendimentos a temperatura de 60ºC (1,504 mg de éster/g de biomassa em 30, 1,518 mg de éster/g de biomassa em 45 minutos e 1,509 mg de éster/g de biomassa em 60 minutos). Na produção de ésteres, a análise fatorial revelou que houve interação entre temperatura e tempo, em média, os melhores rendimentos em número de cetanos C15 e C16 sendo C16 (76,97% em 30ºC, 58,21% em 45Cº, 64,05% em 60Cº e 56,79% em 75CºC) perfil de ésteres obtidos, propícios para a produção de biodiesel. Os resultados obtidos demonstram que a biomassa microalga Chlamydomonas reinhardtii apresenta potencial para produção de biodiesel, quando cultivada nas condições testadas neste estudo.

Large scale biofuel production from microalgae is expected to be integrated with point source CO2 sources, such as coal fired power plants. Flue gas (CO2) integration represents a required nutrient source for accelerated growth while concurrently providing an environmental service. Heavy metals inherent in coal will ultimately be introduced into the culture system. The introduced heavy metals have the potential to bind to microalgae cells, impact growth due to toxicity, and negatively impact the quality of biofuel and other microalgal derived products. Heavy metals As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Sb, Se, Sn, V and Zn, commonly present in coal, were introduced to the microalgae growth medium at a concentration expected from a 7 day growth period using coal flue gas. Experimentation was conducted with Nannochloropsis salina cultivated in photobioreactors at a light intensity of 1000 μmol m-2 s-1. Heavy metals negatively impacted the growth with the average productivity being 0.54 ± 0.28 g L-1 d-1, corresponding to a decrease of 52% in biomass yield compared to control growths. Heavy metal analysis showed significant binding of the majority of the heavy metals to the biomass. A lipid content analysis found a decrease in lipid content from 38.8 ± 0.62% to 31.58 ± 0.50% (percent dry biomass). Control and heavy metal contaminated biomass were processed into biofuel through one of two different in-situ transesterification techniques, either acid-catalyzed or supercritical methanol conversion. The acid-catalyzed conversion resulted in an average crude biofuel production decrease from 0.31 ± 0.03 grams biofuel/gram microalgae for the control algae to 0.28 ± 0.02 grams biofuel/gram microalgae for the heavy metal algae, representing a 9.7% reduction. Supercritical methanol conversion exhibited a similar trend corresponding to a 15.8% reduction. Compared to the control, the total production of biofuel from the contaminated system was decreased by 51% for the acid-catalyzed conversion and 55% for the supercritical methanol conversion. Heavy metal analyses were performed on the biofuel, lipid extracted algae, and other biofuel conversion byproducts. Biochemical methane potential testing was performed on the lipid extracted algae to determine the effect of heavy metals on the generation of biogas. The effects of heavy metals in combination with the effects of acid catalyzed transesterification were found to have a positive effect on the amount of methane produced with an average productivity of 105.89 mL g-COD-1 from the heavy metals contaminated LEA compared to the control microalgae biomass which produced 53.25 mL g-COD-1.

Due to population expiation and the increased awareness of the impact on the environment by wastewater treatment, improved wastewater treatment systems are needed to treat municipal and agricultural wastewater. Treating wastewater with oxygen decreases carbon compounds at the expense of energy to move carbon and oxygen to be in contact with each other. Anaerobic digestion of wastewater can reduce the cost by utilizing microbes to treat high amounts of carbon in wastewater without the need for extensive oxygen requirement. With a proper balance of nutrients, microbes also produce methane, a renewable energy source. It has been suggested that microalgae be utilized to help balance the nitrogen content of wastewater for treatment by microbes. One challenge with the use of algae is the initial breakdown of algae cells. Using a digester with microorganism that can produce methane and decompose algae could enhance production of methane from the digestion of algae. The combination of wastewater, which is high in carbon content relative to nitrogen, with algae, which is high in nitrogen, could provide the balance needed for the microbes to treat wastewater and provide methane. A biomethane potential test was used to compare the ability of two microbe communities, facultative lagoon sediment and anaerobic digester sludge to digest algae. Each microbe community treated dairy, swine, municipal, and petrochemical wastewater augmented with algae and acetate. The ability to degrade augmented wastewater and produce methane was determined by measuring the volume and composition of biogas produced over time. Both treatments were successful in production of methane. Facultative lagoon sediment showed more methane produced per carbon dioxide than anaerobic digester sludge.

As the global population grows, water and food demand also increase. The intensive aquaculture industry has helped to mitigate these problems. In order to make aquaculture sustainable, it is necessary to remove the abundant nutrients produced by fish in the water. In this study, the role of the microalga of Chlorella vulgaris in the Isolated Cultivation of Algal Resources Utilizing Selectivity (ICARUS) membrane photobioreactor was evaluated for nutrient control in the aquaculture system. The production of algal biomass, the removal rate of nutrients, and the impact of microalgae on cultured organisms were monitored during the operation of aquaculture systems. At the end of the experiment, the yield of algae in ICARUS was approximately 344 ± 11.3 mg / L. Compared to the control groups, this production of algae is considered to be low. Likely factors were insufficient indoor light intensity, membrane fouling limiting the mass transfer of nutrients, and improvements still needed for the overall ICARUS prototype design. However, ICARUS can efficiently prevent algae from contamination, and provide pure harvest production for food supplement. It was observed that algae have the ability to help stabilize pH and increase dissolved oxygen for the system. However, in high-density, mixed systems, algae may cause physical damage to fish (e.g., clogging of gills). The high ammonia concentrations produced by fish could be controlled by Chlorella vulgaris since this species of algae prefers ammonia to nitrate. In conjunction with algal growth, aquaculture systems concentration of ammonia was maintained at 0.90±0.16 mg/L. The integration of ICARUS is not only a potentially sustainable option for aquaculture, but also a multipurpose tool for other types of wastewater treatment. An economic analysis for scale-up of the ICARUS system was performed. In summary, this study aimed to develop a new commercial ICARUS photobioreactor which can serve for different types of wastewater systems with a high algal production efficiency and economic benefits.

A prevailing view exists in the literature which suggests that macroalgae growing in slow-flow environments (<4 cm s⁻�) are less productive because of "mass-transfer" limitation compared to fast-flow environments. Macroalgae in slow-flow environments are thought to have thicker diffusion boundary-layers which limit the flux of essential molecules to and from the algal thallus. However nutrient demand of a macroalga can also influence nutrient flux. The main objective of this research was to determine the relative importance of physical (mainstream velocity) and physiological (nutrient demand) factors influencing the growth rate of Adamsiella chauvinii, a small (<20 cm) red algal species, that grows within the benthic boundary-layer in a soft sediment habitat. To establish the influence of water velocity, the growth rate of A. chauvinii was measured in situ each month (March 2003 to March 2004) at three sites with varying degrees of water velocity (slow, intermediate and fast) at which all other environmental parameters (photon flux density, seawater temperature and nutrients) were similar. To determine the metabolic demand and nutrient uptake rate of A. chauvinii, the internal nutrient status (C:N, soluble tissue nitrate, ammonium and phosphate), uptake kinetics (V[max] and K[s]) and nutrient uptake rate at a range of mainstream velocities were also determined on a seasonal basis. The hydrodynamic environment around A. chauvinii canopies was characterised in situ and compared with controlled laboratory experiments. Growth rates of Adamsiella chauvinii thalli at the slow-flow site were significantly lower in winter (June) to summer (February) than the intermediate- and fast-flow sites, while in autumn growth rates were similar between sites. However, A. chauvinii at the slow-flow site had similar or higher tissue N content compared to thalli at the other two sites during winter, spring and summer suggesting that growth rates of A. chauvinii were not mass-transfer limited. Nitrogen uptake rates of A. chauvinii were similar between sites in summer and winter, however uptake rates were lower in summer compared to winter even though thalli were nitrogen limited in summer. Water velocity had no effect on nitrate uptake in either summer or winter and uptake of ammonium increased with increasing water velocity during summer only. Two hydrodynamically different environments were distinguished over a canopy of A. chauvinii, with both the laboratory and field velocity profiles in good agreement with each other. In the top half of the canopy, the Turbulent Kinetic Energy (TKE) and Reynolds stresses were greatest while in the bottom half of the canopy flow rates were less than 90 % of mainstream velocity (< 1 cm s⁻�). When considered together, the influence of water velocity on the growth rates of A. chauvinii was not completely clear. Results suggest that mainstream velocity had little influence on nutrient availability to A. chauvinii because of the unique hydrodynamic environment created by the canopy. Nutrients, especially ammonium and phosphate, derived from the sediment and invertebrates, may provide enough nitrogen and phosphate to saturate the metabolic demand of Adamsiella chauvinii, consequently, A. chauvinii is well adapted to this soft-sediment environment.

This thesis examines the toxicity of the herbicide glyphosate (two formulations ¡V technical grade and Roundup Biactive RB) and the insecticide chlorpyrifos CPF to a model freshwater food chain of a producer and consumer. The importance of studying the toxicity of low (environmentally realistic) concentrations of pesticides to non-target organisms is highlighted. An extensive literature review on the toxicity of glyphosate and chlorpyrifos to aquatic organisms is provided. The requirements for the maintenance of algal (Chlorella vulgaris, Chlorella pyrenoidosa and Pseudokirchneriella subcapitata) and Daphnia carinata cultures are discussed. The effects of two formulations of the herbicide glyphosate (technical grade and Roundup Biactive„µ) and the insecticide chlorpyrifos on the growth of Chlorella pyrenoidosa and Pseudokirchneriella subcapitata were studied, and the EC50 values determined. Hormesis was observed when P. subcapitata was exposed to concentrations of Roundup equal to 7% and 4% of its EC50 respectively. When exposed to chlorpyrifos concentrations 0.3-5 ƒÝg/L, hormesis was observed for both algal species with a maximum at 0.06% of EC50. The effects of sublethal concentrations of chlorpyrifos on population characteristics of Daphnia carinata were investigated in multiple-generation toxicity testing using individual culture. Exposure to chlorpyrifos affected survival and fecundity of animals in the first generation. In the second generation the most affected endpoint was time to the first brood with an indication of hormesis. LC50 tests were then conducted using animals of the third generation from each of the exposures in individual tests. Results of testing the third generation showed a constant significant decline in LC50 in the order of control daphnids through to ¡¥0.1 LC50¡¦ pre-exposed daphnids. The same experimental protocol was used in testing of glyphosate (technical grade and Roundup Biactive). Glyphosate was tested in two different media: sea salt solution and M4 medium, while Roundup Biactive was tested in M4 medium. Results indicated that glyphosate and Roundup Biactive had low toxicity to Daphnia. Hormesis was evident in sea salt medium exposures in the first and second generations of daphnids with glyphosate. When exposed to glyphosate and Roundup Biactive in M4 medium animals showed no indication of hormesis. It is hypothesized that glyphosate may have compensated for the lack of microelements in the sea salt medium, and possible mechanisms discussed.The modifying effect of glyphosate on the toxicity of cadmium to Daphnia carinata was studied using the same experimental design. Low concentrations of Roundup Biactive reduced the toxicity of cadmium, and the performance of daphnia was enhanced in terms of animal size, survival, fecundity, and the rate of natural increase in both generations in the presence of glyphosate. However when the third generation was tested for their sensitivity to Cd in the 48-h LC50 experiments there was no difference between RB-free and RB-spiked treatments in pair wise comparisons, indicating that no adaptation mechanisms were involved in the enhancement. The implications of these observed effects for environmental freshwater food chains subjected to pesticide exposure are discussed and recommendations on modifying pesticide use are provided.

Résumé : Le travail réalisé au cours de cette thèse traite de la mise au point de méthodes d’estimation d’état robuste, avec deux domaines d’application en ligne de mire. Le premier concerne le positionnement sécuritaire en transport. L’objectif est de fournir la position et la vitesse du véhicule sous la forme d’intervalles avec un grand degré de confiance. Le second concerne la synthèse de capteurs logiciels pour les bioprocédés, et en particulier la reconstruction des concentrations de composants réactionnels à partir d’un nombre limité de mesures et d’un modèle mathématique interprétant le comportement dynamique de ces composants. L’objectif principal est de concevoir des algorithmes qui puissent fournir des estimations acceptables en dépit des incertitudes provenant de la mauvaise connaissance du système comme les incertitudes sur les paramètres du modèle ou les incertitudes de mesures. Dans ce contexte, plusieurs algorithmes ont été étudiés et mis au point. Ainsi, dans le cadre du positionnement de véhicule, la recherche s’est dirigée vers les méthodes robustes Hinfini et les méthodes par intervalles. Les méthodes Hinfini sont des méthodes linéaires prenant en compte une incertitude dans la modélisation et réalisant une optimisation min-max, c’est-à-dire minimisant une fonction de coût qui représente la pire situation compte tenu des incertitudes paramétriques. La contribution de ce travail concerne l’extension à des modèles faiblement non linéaires et l’utilisation d’une fenêtre glissante pour faire face à des mesures asynchrones. Les méthodes par intervalles développées ont pour but de calculer les couloirs de confiance des variables position et vitesse en se basant sur la combinaison d’intervalles issus des capteurs d’une part et sur l’utilisation conjointe d’un modèle dynamique et cinématique du véhicule d’autre part. Dans le cadre des capteurs logiciels pour bioprocédés, trois familles de méthodes ont été étudiées: le filtrage particulaire, les méthodes par intervalles et le filtrage par horizon glissant. Le filtrage particulaire est basé sur des méthodes de Monte-Carlo pour estimer la densité de probabilité conditionnelle de l’état connaissant les mesures. Un de ses principaux inconvénients est sa sensibilité aux erreurs paramétriques. La méthode développée s’applique aux bioprocédés et profite de la structure particulière des modèles pour proposer une version du filtrage particulaire robuste aux incertitudes des paramètres cinétiques. Des méthodes d’estimation par intervalles sont adaptées à la situation où les mesures sont disponibles à des instants discrets, avec une faible fréquence d’échantillonnage, en développant des prédicteurs appropriés. L’utilisation d’un faisceau de prédicteurs grâce à des transformations d’état et le couplage entre les prédicteurs avec des réinitialisations fréquentes permettent d’améliorer les résultats d’estimation. Enfin, une méthode basée sur le filtre à horizon glissant est étudiée en effectuant une optimisation min-max : la meilleure condition initiale est reconstruite pour le plus mauvais modèle. Des solutions sont aussi proposées pour minimiser la quantité de calculs. Pour conclure, les méthodes et résultats obtenus constituent un ensemble d’améliorations dans le cadre de la mise au point d’algorithmes robustes vis-à-vis des incertitudes. Selon les applications et les objectifs fixés, telle ou telle famille de méthodes sera privilégiée. Cependant, dans un souci de robustesse, il est souvent utile de fournir les estimations sous forme d’intervalles auxquels est associé un niveau de confiance lié aux conditions de l’estimation. C’est pourquoi, une des méthodes les plus adaptées aux objectifs de robustesse est représentée par les méthodes par intervalles de confiance et leur développement constituera un point de recherche futur. __________________________________________ Abstract : This thesis work is about the synthesis of robust state estimation methods applied to two different domains. The first area is dedicated to the safe positioning in transport. The objective is to compute the vehicle position and velocity by intervals with a great confidence level. The second area is devoted to the software sensor design in bioprocess applications. The component concentrations are estimated from a limited number of measurements and a mathematical model describing the dynamical behavior of the system. The main interest is to design algorithms which achieve estimation performance and take uncertainties into account coming from the model parameters and the measurement errors. In this context, several algorithms have been studied and designed. Concerning the vehicle positioning, the research activities have led to robust Hinfinity methods and interval estimation methods. The robust Hinfinity methods use a linear model taking model uncertainty into account and perform a min-max optimization, minimizing a cost function which describes the worst-case configuration. The contribution in this domain is an extension to some systems with a nonlinear model and the use of a receding time window facing with asynchronous data. The developed interval algorithms compute confidence intervals of the vehicle velocity and position. They use interval combinations by union and intersection operations obtained from sensors along with kinematic and dynamic models. In the context of bioprocesses, three families of state estimation methods have been investigated: particle filtering, interval methods and moving-horizon filtering. The particle filtering is based on Monte-Carlo drawings to estimate the posterior probability density function of the state variables knowing the measurements. A major drawback is its sensitivity to model uncertainties. The proposed algorithm is dedicated to bioprocess applications and takes advantage of the characteristic structure of the models to design an alternative version of the particle filter which is robust to uncertainties in the kinetic terms. Moreover, interval observers are designed in the context of bioprocesses. The objective is to extend the existing methods to discrete-time measurements by developing interval predictors. The use of a bundle of interval predictors thanks to state transformations and the use of the predictor coupling with reinitializations improve significantly the estimation performance. Finally, a moving-horizon filter is designed, based on a min-max optimization problem. The best initial conditions are generated from the model using the worst parameter configuration. Furthermore, additional solutions have been provided to reduce the computational cost. To conclude, the developed algorithms and related results can be seen as improvements in the design of estimation methods which are robust to uncertainties. According to the application and the objectives, a family may be favored. However, in order to satisfy some robustness criteria, an interval is preferred along with a measure of the confidence level describing the conditions of the estimation. That is why, the development of confidence interval observers represents an important topic in the future fields of investigation.

When cultivated properly in bioreactors, microalgae have been found to produce vast amounts of biomass. In the case of diatom cultivation where the organisms will fall out of suspension quite easily, paddle wheels or pumps are the primary means to maintain the necessary velocity in the raceway. This study will focus on the potentially harmful shear stress these devices may impart onto the organisms. The system used to impart shear stress to a diatom culture was a cone and plate viscometer. Cells were counted using a fluorescein diacetate staining method with a fluorescent and brightfield microscope. Under the white light all cells were visible while only the healthy cells were visible under fluorescent light. The sample was exposed to shear stress with the cone and plate viscometer at 6 Pascals for 10 minutes and compared against a non-sheared sample. For each sample, 5 pairs of white and fluorescent light images were captured, counted, and averaged. A non-sheared sample was paired with a sheared sample to calculate the decrease in cell viability. The slope was calculated from the plot of shear stress and cell viability for 9 strains. In each case shear stress resulted in a significant decrease in cell viability; however, there was no statistical difference between strains. While effective, this method would be impractical for a commercial algae cultivation facility as the viscometer in this study costs approximately $100,000. Therefore, tests were performed to determine if a rotary mixer could be substituted for the viscometer. The hypothesis was that the cell damage was a product of shear stress and exposure time. For the viscometer test, the shear exposure was 3600 Pa s. Two rotational mixer tests were performed, one at 1250 RPM for 7 hours and one at 313 RPM for 28 hours, providing the same 3600 Pa s shear exposure. After staining, cell viability decreased 35.62% and 11.07% in the 1250 RPM and 313 RPM test, respectively. This difference was significant compared to the 6.04% decrease in the viscometer test. The increased cell damage was attributed to turbulence in the mixer tests and the basis for further study.

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.