The role of energy as a stimulant for economic growth and environmental sustainabilityof any nation has made the focus on green fuels, including fermentative hydrogen (bioH2) andmethane (bioCH4), to be a priority for the World’s policy makers. Nigeria, as the most populousAfrican country, with worsening energy crisis, can benefit from the introduction of the bioH2 andbioCH4 technologies into the country’s energy mix, since such technologies have the potential ofgenerating energy from organic wastes such as fruit waste.Fruit waste was studied in detail in this work because of its great economic andenvironmental potential, as large quantities of the wastes (10–65% of raw fruit) are generatedfrom fruit consumption and processing. Meanwhile, bioH2 and bioCH4 productions involvinganaerobic microorganisms in direct contact with organic wastes have been observed to result insubstrate and product inhibitions, which reduce the gas yields and limit the application of thetechnologies on an industrial scale. For example, in this study, the first experimental work todetermine the effects of hydraulic retention times and fruit mixing on bioH2 production fromsingle and mixed fruits revealed the highest cumulative bioH2 yield to be equivalent to 30% ofthe theoretical yield. However, combining the fermentation process with the application ofmembrane encapsulated cells and membrane separation techniques, respectively, could reducesubstrate and product inhibitions of the microorganisms. This study, therefore, focused on theapplication of membrane techniques to enhance the yields of bioH2 and bioCH4 productions fromthe organic wastes.The second experimental work which focused on reduction of substrate inhibition,involved the investigation of the effects of the PVDF membrane encapsulation techniques on thebioH2 and bioCH4 productions from nutrient media with limonene, myrcene, octanol and hexanalas fruit flavours. The results showed that membrane encapsulated cells produced bioCH4 fasterand lasted longer, compared to free cells in limonene. Also, about 60% membrane protectiveeffect against myrcene, octanol and hexanal inhibitions was obtained. Regarding bioH2production, membrane encapsulated cells, compared to free cells, produced higher average dailyyields of 94, 30 and 77% with hexanal, myrcene and octanol as flavours, respectively. The finalpart of the study, which was aimed at reducing product inhibition, involved the study of theeffects of membrane permeation of volatile fatty acids (VFAs) on the bioreactor hydrodynamicsin relation to bioH2 production. The investigation revealed that low transmembrane pressure of104Pa was required to achieve a 3L h-1m-2 critical flux with reversible fouling mainly due to cakelayer formation, and bioH2 production was also observed to restart after VFAs removal.The results from this study suggest that membrane-based techniques could improve bioH2and bioCH4 productions from fermentation media with substrate and product inhibitions.

The degradation of toxic and volatile contaminants in aqueous streams is considered a challenge using conventional bioremediation strategies. At moderate concentrations, toxic contaminants induce microbial inhibition, which results in an overall decrease of reaction rates. On the other hand, volatile compounds are often stripped out of solution into the atmosphere during aeration in conventional wastewater treatments, and are not treated. The addition of a second non-aqueous phase with affinities for the contaminants can reduce aqueous concentrations to sub-inhibitory levels and also decrease contaminant volatilization, while still allowing controlled release of contaminants back to the microbial population; such systems have been denoted as Two Phase Partitioning Bioreactor (TPPB). The current work examined and compared the performance of solid-liquid TPPB to a liquid-liquid TPPB and a single phase system. The systems were compared in the simultaneous degradation of phenol and butyl acetate, two substrates known for their relatively high levels of toxicity and volatility, respectively. The solid-liquid TPPB, using 2 polymers selected heuristically, showed an improvement of 40 and 54 % in phenol degradation rates compared to the single phase and the liquid-liquid systems. Additionally, the solid-liquid system presented a 55 and 11 % enhancement in the amount of butyl acetate degraded. At higher initial substrate concentration the solid-liquid TPPB showed an improvement in the phenol degradation rate and the amount of butyl acetate degraded of 44 and 94 % respectively, compared to the single phase system. In order to rationalize polymer screening for solid-liquid TPPBs, selection criteria based on first principles were developed, and were based on consideration of polymer accessibility and polymer-solute thermodynamic affinity. Polymer accessibility was evaluated by considering glass transition temperature (Tg) and degree of crystallinity, while polymer-solute thermodynamic affinity was assessed using three different methods, Hildebrand solubility parameters, Hansen iii Solubility Parameters (HSP) and activity coefficients at infinite dilution. It was found that the HSP method gave the best trends and its predictions had better agreement with the experimental results. Consequent biodegradation experiments with a single, rationally selected polymer, and a mixture of waste polymers, demonstrated the superior performance of rational selected polymers. / Thesis (Master, Chemical Engineering) -- Queen's University, 2013-05-02 16:24:39.655

Extraction of bitumen from Alberta oil sands produces 2 to 4 barrels of aqueous tailings per barrel of crude oil. Oil sands process water (OSPW) contains naphthenic acids (NAs), a complex mixture of carboxylic acids of the form CnH2n+ZOx that are persistent and toxic to aquatic organisms. Previous studies have demonstrated that aerobic biodegradation reduces NA concentrations and OSPW toxicity; however, treatment times are long. The objective of this study was to evaluate the feasibility of an immobilized soil bioreactor (ISBR) for treatment of NAs in OSPW and to determine the role of ammonium and ammonium oxidizing bacteria (AOB) in NA removal. ISBRs have been used to successfully remediate water contaminated with pollutants such as pentachlorophenol and petroleum hydrocarbons. A system of two ISBRs was operated continuously for over 2 years with OSPW as the sole source of carbon. Removal levels of 30-40% were consistently achieved at a residence time of 7 days, a significant improvement compared to half-lives of 44 to 240 days reported in the literature. However, similar to biodegradation experiments in the literature, a significant portion (~60%) of the NAs was not degraded. The role of AOB in NA removal was investigated by decreasing ammonium concentration and inhibiting AOB activity with allylthiourea, neither of which significantly affected removal, indicating that AOB did not enhance NA removal. Furthermore, high AOB populations actually inhibited the removal of a simple NA surrogate. Therefore, a moderate ammonium concentration of 0.3 g/L is recommended. NA degradation occurred with nitrate as the sole nitrogen source, however, removal levels were lower than those achieved with ammonium. Exploratory studies involving ozonation or biostimulation were conducted with the aim of increasing NA removal. Ozonation decreased NA concentration by 94% and total organic carbon (TOC) by 6%. Subsequent ISBR treatment removed ~30% of the remaining TOC. Addition of a NA surrogate increased heterotrophic NA-degrading populations due to the increase in available carbon, resulting in a significant increase in NA removal levels. However, use of a surrogate may result in a population that is only adapted to degradation of the NA surrogate. / Thesis (Master, Chemical Engineering) -- Queen's University, 2013-06-20 14:53:47.498

With the aim of mitigating membrane fouling, an electrocoagulation (EC) based electrokinetic membrane bioreactor (EMBR) was developed and operated with real municipal wastewater under low temperatures. Both batch tests and continuous EMBR experiments demonstrated the significant advantages in membrane fouling reduction over the conventional antifouling strategies, ushering its potential applications as an attractive hybrid MBR technology for decentralized wastewater treatment in remote cold regions. The main research observations and findings could be summarized as follows: (1). Effective membrane fouling mitigation at low temperatures was due to destruction of extracellular polymeric substances (EPS) and subsequent reduction of the biocake resistance. The transmembrane pressure (TMP) increased at a much slower rate in EMBR and the filtration resistance was about one third of the control MBR prior to chemical cleaning cycle; (2). A new membrane parameter, the specific fouling rate (SFR) was proposed, relating the fouling rate with permeate flux and temperature-dependent viscosity. Pore clogging and biocake resistances were quantified for the first time with the same membrane module and operating conditions as in regular MBR, rather than resorting to the use of batch filtration setups; (3). The floc size in EMBR did not increase as a result of the air scouring shear force and decrease in the extracellular polymeric substances (EPS); (4). When current intensity was less than 0.2 A, polarity reversal had minimal impact on electrode passivation reduction due to insignificant hydrogen yield, however, if current intensity was above 0.2 A, frequent polarity reversal (< 5 min per cycle) was detrimental to electrode passivation if no sufficient mixing was provided; (5). Viability of the microorganisms in the EMBR system was found to be dependent on duration of the current application and current density. The bacterial viability was not significantly affected when the applied current density was less than 6.2 A/m2; (6). Significant abiotic ammonification was found in electrocoagulation (EC). DO in the treated liquid was depleted within an hour, under the anaerobic condition in EC, nitrate was chemometrically reduced to ammonium following a two-step first order reaction kinetics. Aeration (DO > 2 mg/L) was shown effective in suppressing abiotic ammonification; (7). Magnetic resonance imaging (MRI) technology was used for the first time as an in-situ non-invasive imaging tool to observe membrane fouling status in an EMBR. / October 2016

Membrane bioreactors (MBRs) show great promise for productivity improvement and energy conservation in conventional bioprocesses for wastewater reclamation. In order to attain high productivity in a bioprocess, it is crucial to retain the microorganisms in the bioreactors by preventing wash out. This enables recycling of the microorganisms, and is consequently saving energy. The main feature of MBRs is their permeable membranes, acting as a limitative interface between the medium and the microorganisms. Permeation of nutrients and metabolites through the membranes is thus dependent on the membrane characteristics, i.e. porosity, hydrophilicity,and polarity. The present thesis introduces membranes for MBRs to be used in a continuous feeding process, designed in the form of robust, durable, and semi-hydrophilic films that constitute an effective barrier for the microorganisms, while permitting passage of nutrients and metabolites. Polyamide 46 (polytetramethylene adipamide), a robust synthetic polymer, holds the desired capabilities, with the exception of porosity and hydrophilicity. In order to achieve adequate porosity and hydrophilicity, bulk functionalization of polyamide 46 with different reagents was performed. These procedures changed the configuration from dense planar to spherical, resulting in increased porosity. Hydroxyethylation of the changed membranes increased the surface tension from 11.2 to 44.6 mJ/m2. The enhanced hydrophilicity of PA 46 resulted in high productivity of biogas formation in a compact MBR, due to diminished biofouling. Copolymerization of hydrophilized polyamide 46 with hydroxymethyl 3,4-ethylenedioxythiophene revealed electroconductivity and hydrophilic properties, adequate for use in MBRs. To find either the maximal pH stability or the surface charge of the membranes having undergone carboxymethylation, polarity and the isoelectric point (pI) of the treated membranes were studied by means of a Zeta analyzer. The hydroxylated PA 46 was finally employed in a multilayer membrane bioreactor and compared with hydrophobic polyamide and PVDF membranes. The resulting biogas production showed that the hydroxylated PA 46 membrane was, after 18 days without regeneration, fully comparable with PVDF membranes.

Mechanical stimulation, in combination with biochemical factors, is likely to be essential to the appropriate function of stem cells and the development of tissue engineered constructs for orthopaedic and other uses. A multi-axial bioreactor was designed and built by Bose ElectroForce to simulate physiologically relevant loading conditions of the intervertebral disc (IVD), including axial compression, hydrostatic pressure and perfusion flow to multiple constructs under the control of a software program. This research optimises the design and configuration of the perfusion system of the bioreactor and presents results of preliminary experimental work on the combined effects of axial compression and perfusion on the viability of mesenchymal stem cells encapsulated in alginate hydrogels and the ability of the cells to produce extracellular matrix (ECM). The results of this thesis illustrated the power of a design of experiments (DOE) approach as a troubleshooting quality tool. With a modest amount of effort, we have gained a better understanding of the perfusion process of the tri-axial bioreactor, improved operational procedures and reduced variation in the process. Furthermore, removing unnecessary tubing lengths, equipment and fittings has made cost savings. The steady flow energy equation (SFEE) was used to develop a numerical analysis framework that provides an insight into the balance between velocity, elevation and friction in the flow system. The pressure predictions agreed well with experimental data, thus validating the SFEE for fluid analysis in the bioreactor system. The numerical predictions can be used to estimate the pressures around the three-dimensional constructs with a given arrangement of the tubing and components of the bioreactor. The system can potentially support long-term cultures of cell-seeded constructs in controlled environmental conditions found in vivo to study the mechanobiology of nucleus pulposus tissue engineering and the aetiology of IVD degeneration. However, dynamic compression and perfusion with associated hydrostatic pressurization of culture medium resulted in significant loss of cell viability compared to the unstimulated controls. Due to a large number of factors affecting cell behaviour in the tri-axial bioreactor system, it is difficult to identify the exact parameters influencing the observed cell response. A strategy that could help to distinguish the effects of mechanical stimuli and specific physiochemical factors should combine experiments with mathematical modelling approaches, and use the sensing incorporated in the bioreactor design and process-control systems to monitor and control specific culture parameters. Optimisation of the cell passage and cell seeding density were identified as key areas to improve the production of GAG in future studies; since the production of ECM was not observed in both static and dynamic cultures. Further studies could also attempt to use other hydrogel scaffolds, such as agarose, which has been widely used in cartilage tissue engineering studies and hyaluronic acid - a component of the nucleus pulposus ECM.

A relatively new concept of Municipal Solid Waste treatment is known as bioreactor landfill technology. Bioreactor landfills are sanitary landfills that use microbiological processes purposefully to transform and stabilize the biodegradable organic waste constituents in a shorter period of time. One of the most popular types of bioreactor landfills is the landfill with leachate recirculation. However, it is observed that ammonia rapidly accumulates in landfills that recirculate leachate and may be the component that limits the potential to discharge excess leachate to the environment. In the facultative landfill, leachate is nitrified biologically using an on-site treatment plant and converted by denitrifying bacteria to nitrogen gas, a harmless end-product. In this research, three pilot-plant scale lysimeters are used in a comparative evaluation of the effect of recirculating treated and untreated leachate on waste stabilization rates. The three lysimeters are filled with waste prepared with identical composition. One is being operated as a facultative bioreactor landfill with external leachate pre-treatment prior to recirculation, the second is being operated as an anaerobic bioreactor landfill with straight raw leachate recirculation, and the third one is the control unit and operated as a conventional landfill. Apart from environmental restrictions, geotechnical constraints are also imposed on new sanitary landfills. The scarcity of new potential disposal areas imposes higher and higher landfills, in order to utilize the maximum capacity ofthose areas. In this context, the knowledge of the compressibility of waste landfills represents a powerful tool to search for alternatives for optimization of disposal areas and new solid waste disposal technologies. This dissertation deals with and discusses the environmental and geotechnical aspects of municipal solid waste landfills. In the Environmental Engineering area, it compares the quality of the leachate and gas generated in the three lysimeters and discusses the transfer of the technology studied through lysimeters to procedures for full-scale operation. In the geotechnical area, this dissertation discusses the compressibility properties of the waste and provides a state-of-the-art review of MSW compressibility studies. It also evaluates the compressibility of MSW landfills for immediate and long-term settlements and proposes a new model for compressibility of waste landfills.

Tissue engineered blood vessels (TEBVs) have great potential as tools for disease modeling and drug screening. However, existing methods for fabricating TEBVs create homogenous tissue tubes, which may not be conducive to modeling focal vascular diseases such as intimal hyperplasia or aneurysm. In contrast, our lab has a unique modular system for fabricating TEBVs. Smooth muscle cells (SMCs) are seeded into an annular agarose mold, where they aggregate into vascular tissue rings, which can be stacked and fused into small diameter TEBVs. Our goal is to create a platform technology that may be used for fabricating focal vascular disease models, such as intimal hyperplasia. Because tubes are fabricated from individual ring units, each ring can potentially be customized, enabling the creation of focal changes or regions of disease along the tube length. In these studies, we first demonstrated our ability to modulate cell phenotype within individual SMC ring units using incorporated growth factor-loaded degradable gelatin microspheres. Next, we evaluated fusion of ring subunits to form composite tissue tubes, and demonstrated that cells retain their spatial positioning within individual rings during fusion. By incorporating electrospun polycaprolactone cannulation cuffs at each end, tubes were mounted on bioreactors after only 7 days of fusion to impart luminal medium flow for 7 days at a physiological shear stress of 12 dyne/cm2. We then created focal heterogeneities along the tube length by fusing microsphere-containing rings in the central region of the tube between rings without microspheres. In the future, microspheres may be used to deliver growth factors to this localized region of microsphere incorporation and induce disease phenotypes. Due to the challenges of working with primary human SMCs, we next evaluated human mesenchymal stem cells (hMSCs) as an alternative cell source to generate vascular SMCs. We evaluated the effects of microsphere-mediated platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), and transforming growth factor beta-1 (TGF-β1) delivery on ring thickness, proliferation, and contractile protein expression over a 14 day period. Finally, we created a structurally distinct region of smooth muscle within tissue tubes by fusing human aortic SMCs in a central region between hMSC rings. In summary, we developed a platform technology for creating modular tubular tissues that may be further developed into an in vitro intimal hyperplasia model. It may also be modified to model other focal vascular diseases, such as aneurysm, or to create other types of multi-tissue tubular structures, such as trachea.

Os polissacarídeos possuem diversas aplicações industriais devido a sua ampla variedade de propriedades físico-químicas. Além desses empregos tradicionais, pesquisas mais recentes estão impulsionando o uso de polissacarídeos para novas aplicações, principalmente na área de terapia farmacêutica. Com o objetivo de ampliar a produção industrial de polissacarídeos microbianos, pesquisas têm se concentrado nos exopolissacarídeos (EPS), que apresentam produtividade elevada e processos de extração e purificação mais simples quando comparados aos polímeros tradicionais. A lasiodiplodana, uma ?-(1->6)-D-glucana, é um EPS produzido por Lasiodiplodia theobromae, um fungo filamentoso característico de regiões tropicais e patógeno de mais de 500 espécies vegetais. Na produção de EPS observa-se que, como resultado do crescimento e da produção do biopolímero, o meio de cultivo torna-se mais viscoso, tornando difícil a manutenção da homogeneidade no biorreator e, consequentemente, prejudicando a transferência de massa e oxigênio no meio. Nos cultivos em que ocorrem mudanças na reologia do meio e nos quais são utilizados fungos filamentosos, sensíveis ao cisalhamento, o uso de biorreatores convencionais, como o Stirred Tank Reactor (STR) e o Airlift, não é adequado. Nesse contexto, no presente estudo tem-se como objetivo estudar a cinética de crescimento, consumo de substrato e produção de EPS pelo fungo filamentoso Lasiodiplodia theobromae, a partir de glicose e de glicerol, no Biorreator Agitado e Aerado de Baixo Cisalhamento (BAABC) e compará-lo com o STR. No estudo inicial, realizado em frascos agitados, a maior produção de EPS (6,49 ± 0,03 g/L) foi alcançada pelo ensaio G, que continha a maior concentração glicose testada (50 g/L) e a menor concentração de extrato de levedura (3 g/L). Quando se utilizou glicerol como fonte de carbono, a maior produção de EPS (3,39 ± 0,06 g/L) foi observada no Ensaio O, que continha 30 g/L de glicerol e a maior concentração de fonte de nitrogênio testada (12 g/L de extrato de levedura). Nos ensaios em biorreatores, quando utilizou-se glicose como fonte de carbono observou-se que o BAABC com controle de temperatura proporcionou uma produção de 3,17 ± 0,16 g/L de EPS, concentração inferior à obtida em frascos, porém bastante superior à obtida no biorreator STR (0,70 ± 0,12 g/L). Para os meios contendo glicerol, o biorreator STR proporcionou uma produção de EPS de 3,02 ± 0,19 g/L, enquanto no BAABC a concentração de EPS obtida foi muito menor (1,45 ± 0,25 g/L). Analisando-se, porém, a concentração máxima de biomassa obtida (28,86 ± 1,46 g/L) bem como os parâmetros cinéticos relacionados (YX/S e QX), nota-se que a produção de biomassa foi muito superior e mais eficiente no BAABC. Apesar das diferenças morfológicas visualmente observadas e confirmadas por meio de microscopia óptica nos EPS obtidos a partir de glicose e de glicerol, as análises de Difração de raios-X e Espectroscopia de absorção na região do infravermelho com Transformada de Fourier permitiram evidenciar a similaridade estrutural entre ambos os EPS. / Polysaccharides have several industrial applications because of their wide variety of physicochemical properties. In addition to these traditional applications, recent research is driving the use of polysaccharides towards new applications, especially in the field of pharmaceutical therapy. With the aim of increasing the industrial production of microbial polysaccharides, research has focused on exopolysaccharides (EPS), which have high productivity and simpler extraction and purification processes when compared to traditional polymers. Lasiodiplodana, a ?-(1->6)-D-glucan, is an EPS produced by Lasiodiplodia theobromae, a filamentous fungus characteristic of tropical areas and pathogenic of more than 500 plant species. In EPS production, as a result of the growth and biopolymer production, the culture medium becomes more viscous, making it difficult to maintain homogeneity inside the bioreactor and, consequently, harming the transfer of mass and oxygen in the medium. In cultures where changes in media rheology occur and in which shear-sensitive filamentous fungi are used, the use of conventional bioreactors, such as Stirred Tank Reactor (STR) and Airlift, is not appropriate. In this context, the objective of the present study was to study the growth kinetics, substrate consumption and EPS production by the Lasiodiplodia theobromae filamentous fungus, from glucose and glycerol, in the Low-Shear Aerated-Agitated Bioreactor (LSAAB) and compare it with the STR. In the initial study, conducted in shaken flasks, the highest EPS production (6.49 ± 0.03 g/L) was achieved by the G test, which contained the highest glucose concentration tested (50 g/L) and the lowest concentration of yeast extract (3 g/L). When glycerol was used as the carbon source, the highest EPS yield (3.39 ± 0.06 g/L) was observed in Test O, which contained 30 g/L glycerol and the highest concentration of nitrogen source tested (12 g/L of yeast extract). In the bioreactor trials, when glucose was used as the carbon source, it was observed that the LSAAB with temperature control provided a production of 3.17 ± 0.16 g/L of EPS, a concentration lower than that obtained in flasks, but rather higher than that obtained in the STR bioreactor (0.70 ± 0.12 g/L). For the glycerol-containing media, the STR bioreactor produced 3.02 ± 0.19 g/L of EPS, while in LSAAB the EPS concentration obtained was much lower (1.45 ± 0.25 g/L). However, analyzing the maximum concentration of biomass obtained (28.86 ± 1.46 g/L) as well as the related kinetic parameters (YX/S and QX), biomass production was much higher and more efficient in LSAAB. In spite of the visually observed morphological differences confirmed by optical microscopy in EPS obtained from glucose and glycerol, X-ray diffraction and absorption spectroscopy analyzes in the infrared region with Fourier transform showed the structural similarity between both EPS.

Tissue engineering offers a potential alternative therapy to overcome the limitations of organ transplantation, by employing biomaterials as scaffold for cell growth. For example, poly-lactic-co-glycolic acid (PLGA) is a synthetic biomaterial widely used in tissue engineering. However, the hydrophobicity of PLGA results in scaffolds that are poorly wettable, and which, therefore, possess poor mass transfer properties for the delivery of nutrients and the removal of waste. The present work aimed to develop more hydrophilic PLGA scaffolds, specifically hollow fibre membranes, within a bioreactor system, which enables co-culture of cells in order to direct stem cell differentiation. Large quantities of costly growth factors are required over long periods for stem cell differentiation. Therefore, this project also aimed to use a cell line as a “factory” for the inexpensive, in situ growth factor production. Hollow fibres were fabricated by wet spinning and a hydrophilic polymer, polyvinyl alcohol (PVA), was added to the PLGA solution at three different concentrations (1.25, 2.5, 5% w/w) in order to obtain a more hydrophilic membrane. Results indicated that 5% PVA-PLGA hollow fibres were the only membranes which allowed permeation of water, BSA and cell-secreted hepatocyte growth factor (HGF), thus indicating that they are the most suitable membranes for use in bioreactor devices. However, these membranes failed to improve cell-attachment. Cell secreted HGF was shown to be more stable in a dynamic culture environment than commercial HGF, thus suggesting its suitability for applications in bioreactor devices. However, using both commercial and cell-secreted HGF, mesenchymal stem cell differentiation was unsuccessful. In conclusion, this work has developed a hollow fibre membrane which is more permeable to water and proteins for a higher mass transfer of nutrients, and has realised a model system for the inexpensive production of growth factors for use in bioreactor devices and the differentiation of stem cells.