Aerobic granular sludge has been regarded as a promising alternative to the conventional activated sludge which has been used for a century in that granular sludge offers advantages in high biomass retention, fast sludge-water separation, and small footprint requirement. However, this technology has been rarely applied in continuous flow reactors (CFRs) which are the most common type of bioreactors used in water resource recovery facilities across the world. Hence, the overarching goal of this study is to provide advanced understanding of biogranulation mechanism to enable industrial application of this technology. The lack of long-term stability study in CFRs has restricted its full-scale acceptability. The high settling velocity-based selection pressure has been regarded as the ultimate driving force towards biogranulation in sequential batch reactors (SBRs). In this study, this physical selection pressure was firstly weakened and then eliminated in CFRs to investigate its role in maintaining the long-term structural stability of aerobic granules. Given the fact that implementing settling velocity-based selection pressure only can cultivate biogranules in SBRs but not in completely stirred tank reactors (CSTRs), the essential role of feast/famine conditions was investigated. Seventeen sets of data collected from both literature and this study were analyzed to develop a general understanding of the granulation mechanisms. The outcome indicated that granulation is more sensitive to the feast/famine conditions than to the settling velocity-based selection pressure. The theory was tested in a CFR with 10-CSTR chambers connected in series to provide feast/famine conditions followed by a physical selector separating the slow-settling bioflocs and fast-settling biogranules into feast and famine zones, respectively. Along with successful biogranulation, the startup performance interruption problem inherent in SBRs was also resolved in this innovative design because the sludge loss due to physical washout selection was mitigated by returning bioflocs to the famine zone. Then, a cost-effective engineering strategy was put forward to promote the full-scale application of this advanced technology. With this generalized biogranulation theory, pure culture biogranules with desired functions for high value-added bioproducts were also investigated and achieved for the first time in this study, which paves a new avenue to harnessing granulation technology for intensifying waste valorization bioprocesses. / Doctor of Philosophy / Nowadays, the rapid population growth and unprecedented urbanization are overloading the capacity of many wastewater resource recovery facilities (WRRFs). Therefore, there is a need to develop a cost-effective strategy to upgrade the treatment capacity of existing WRRFs without incurring major capital investment. Because conventional activated sludge comes with loose structure and poor settleability, replacing them with dense aerobic granular sludge offers the opportunity to intensify the capacity of existing WRRF tankage and clarifiers through better retention of high bacterial mass that offers not only a fast pollutant removal rate but also a high water-solids separation rate. The aerobic granulation technology turns traditional activated sludge into granular sludge by inducing microbial cell-to-cell co-aggregation. Although this technology has been developed for more than 20 years, its application in full-scale WRRFs is still limited because majority of WRRFs are constructed with continuous flow reactors in which the aerobic granulation mechanism largely remains unknown. Besides, the long-term stability of aerobic granules in continuous flow reactors also remain unstudied, further constraining the full-scale application of the technology. The sensitivity of aerobic granulation to physical selection and biological selection was analyzed in this study. The results concluded that aerobic granulation is more sensitive to the latter but not to the former. Subsequently, this theory was tested in a novel bioreactor setup that creates feast/famine conditions for biological selection. A physical selector was installed at the end of the bioreactor to separate and return the fast- and slow- settling bioparticles to the feast and famine zones, respectively. This unique reactor design and operational strategy provided an economical approach to retrofitting current WRRFs for achieving treatment capacity upgrading without major infrastructure alternation. It also protected the bioreactor startup performance by enhancing the stability of WRRFs in the future application. Last but not least, this updated understanding of aerobic granulation theory was for the first time extrapolated to and verified with the formation of pure culture biogranules harnessed in this study for value-added bioproduct valorization from waste materials.

Attached growth biological processes offer advantages over traditional water purification technologies through high biomass retention, easy sludge-water separation, multiple multispecies synergies in proximity, resilience to shock loading, low space requirements, and reactor operational flexibility. Traditionally, attached growth refers to biofilms that require abiotic carrying media for bacteria to attach and grow on. While biofilms have been broadly applied in wastewater treatment, its potential for potable reuse or stormwater treatment has not been well studied. The treatment trains of pre-ozonation followed by biologically active filtration (ozone- BAF) is an advanced biofilm technology for potable reuse that can generate high-quality potable water at reduced energy and chemical demands by removing pollutant through three different pathways: oxidation, adsorption, and biodegradation. However, these pathways can result in both desirable and undesirable effects, and the mechanism behind it is still unclear. To understand the mechanisms of various pollutant removal, parallel performance comparisons of ozone-BAF treatment trains with spent and regenerated granular activated carbon (GAC), along with a range of pre-oxidant ozone doses were performed. Another common issue of BAF is the headloss buildup during its operation, which has become a significant energy and maintenance burden at many utilities. Thus, a mathematical model was developed to predict BAF headloss buildup in response to organic removal and nitrification. For stormwater treatment, the feasibility of using biofilms for stormwater biological nitrogen removal (BNR) is still largely unknown, as very limited research effort has been dedicated to this aspect. Thus, a mathematical model was developed to evaluate the potential of using BNR techniques for stormwater nitrogen removal. Aerobic granules are an even more advanced attached growth process, which eliminates the need for abiotic carrying media. So far, aerobic granular sludge is only formed in sequential batch reactors but not in a continuous flow system. Therefore, continuous flow aerobic granulation from traditional activated sludge was investigated and, for the first time, successfully achieved in continuous flow plug-flow bioreactors fed with real municipal wastewater. Besides, the role and critical value of an essential operational parameter, feast/famine ratio, for continuous flow aerobic granulation were determined. / Doctor of Philosophy / Water scarcity and increasing water demand caused by urban population growth and climate change is a reality throughout the world. Thus, process intensification of the current water and wastewater technologies is gaining increasing attention globally. Comparing to traditional water purification technologies, attached growth biological processes offers advantages such as high biomass retention, easy sludge-water separation, multiple multispecies synergies in proximity, resilience to shock loading, small footprint requirement, and reactor operational flexibility. Traditionally, attached growth refers to biofilms that require abiotic carrying media for bacteria to attach and grow on. While biofilms have been broadly applied in wastewater treatment, its potential for potable reuse or stormwater treatment has not been well studied. For potable reuse, the treatment trains of pre-ozonation followed by biologically active filtration (ozone-BAF) is an advanced biofilm technology that can generate high-quality potable water at reduced energy and chemical demands by removing pollutant through different pathways. However, the mechanism behind it is still unclear. To understand the mechanisms of various pollutant removal, parallel performance comparisons of ozone-BAF treatment trains operated with different operational conditions were performed in this dissertation. Another common issue of BAF is the headloss buildup during its operation, which has become a significant energy and maintenance burden at many utilities. Thus, a mathematical model was developed to predict the headloss buildup during BAF operation. For stormwater treatment, the feasibility of using biofilms for stormwater biological nitrogen removal (BNR) is still largely unknown, as very limited research effort has been dedicated to this aspect. Thus, a mathematical model was developed to evaluate the potential of using BNR technique for stormwater. Aerobic granules are an even more advanced attached growth process. However, aerobic granular sludge is so far only formed in sequential batch reactors which are incompatible with the continuous flow nature of most wastewater treatment plants. Therefore, aerobic granulation from traditional activated sludge was investigated and, for the first time, successfully achieved in continuous flow plug-flow bioreactors fed with real municipal wastewater. Besides, the role of an essential operational parameter, feast/famine ratio, for continuous flow aerobic granulation was determined.

Dans ce travail, le traitement des nutriments et plus particulièrement celui de l’azote en procédé de granulation aérobie a été étudié. L’approche expérimentale a dans un premier temps eu pour objet de comparer la stabilité des caractéristiques physiques et microbiologiques d’agrégats développés dans deux réacteurs fonctionnant en alternances de phases anaérobie / aérobie ou anoxie / aérobie. L’opération d’un procédé de granulation aérobie en alternance de phases anoxie / aérobie a favorisé la stabilisation des performances de traitement de l’azote et notamment celle de la nitrification. Le développement d’une boue hybride comprenant une fraction de flocs et de granules a été observé. Dans le but d’évaluer comment le caractère hybride de la boue obtenue en conditions anoxie/aérobie oriente les performances et vitesses de transformation de l’azote, une caractérisation ex-situ des limitations au transfert d’oxygène au sein des flocs, granules et boue hybride a été réalisée par respirométrie. En complément, la localisation des espèces nitrifiantes par la technique d’hybridation fluorescente in situ (FISH) a été réalisée. Les résultats obtenus indiquent que la présence de flocs au sein d’une boue granulaire permet d’augmenter la vitesse de la nitrification, en particulier pour de faibles concentrations en oxygène dissous. D’autre part, il est ici mis en évidence la nécessité de contrôler simultanément le ratio floc/granules et la taille des granules en vue de l’optimisation du traitement de l’azote.Enfin un outil mathématique permettant de décrire les phénomènes réactionnels ayant lieu au sein d’une biomasse hybride a été développé. Celui-ci a été employé dans le but d’optimiser le fonctionnement du procédé hybride par l’évaluation de l’effet des propriétés physiques de la biomasse, et plus particulièrement de la proportion de granules en présence, pouvant conduire à une élimination efficace de l’azote et à l’augmentation de la robustesse du procédé vis-à-vis de diminutions ponctuelles de l’oxygène dissous / In this work, the treatment of nutrients, especially that of nitrogen in aerobic granulation process was studied.The experimental approach has initially been intended to compare the stability and the physical and microbiological characteristics of aggregates developed in two reactors operating in alternating anaerobic / aerobic or anoxic / aerobic conditions. The presence of a pre-anoxic phase promoted the stabilization of nitrogen removal performances and especially those of nitrification. The development of a hybrid sludge process containing a fraction of flocs and granules was observed.In order to evaluate and quantify the influence of the simultaneous presence of flocs and granules in the nitrifying activity of the hybrid sludge developed in the alternating anoxic / aerobic conditions, the nitrification rate and oxygen limitation of flocs, granules and hybrid sludge was assessed using respirometric assays at different dissolved oxygen concentrations. The spatial distribution of nitrifying bacteria was investigated using fluorescence in situ hybridization (FISH). Results indicated that the presence of flocs with granules could increase the rate of nitrification to transitory reductions of aeration. On the other hand, the optimization of nitrogen removal requires the simultaneous control of the floc to granule ratio and granule size.Finally, a mathematical model to describe the reaction phenomena taking place in the hybrid biomass was developed. It was used in order to optimize the operation of the hybrid process through the evaluation of the effect of physical properties of biomass, specifically the ratio of granules and flocs in the reactor that can lead to efficient removal of nitrogen and increase the robustness of the process