After construction upgrades and implementation of ammonia-based aeration control (ABAC), Hampton Roads Sanitation District's Virginia Initiative Plant (VIP) observed a 69 percent decrease in average dissolved oxygen (DO) concentrations, alongside a 53 percent reduction in average effluent total phosphorus (TP) concentrations from 2019 to 2023. This improvement in effluent quality coincided with the elimination of metal salt addition in 2023. Batch tests conducted from 2020 to 2024 indicated higher phosphorus release and aerobic uptake rates at lower DOs, even at higher temperatures, while 16S rRNA amplicon sequencing analysis suggested a community shift toward polyphosphate-accumulating organisms (PAOs). Statistical analysis revealed that low DO operation (DO concentrations below 1 mg O2/L) did not negatively impact effluent TP concentrations and were positively correlated with increased PAO abundance. High rates of denitrification fueled by internally stored carbon in the post-anoxic zone were found to co-occur with elevated PAO activity, and subsequent batch tests indicated post-anoxic phosphorus uptake rates ranging from 3 to 40 percent of the aerobic phosphorus uptake rates. Removing the aerobic phase in batch tests increased both anoxic phosphorus uptake and denitrification utilizing internally stored carbon. This emergence of post anoxic phosphorus uptake capacity is potentially attributable to the reduction in DO concentrations. The reduction in average aerobic SRT from 8.5 ± 0.4 days in 2021 to 5.7 ± 0.1 days in 2023 was significantly correlated with improved effluent phosphorus quality. An aerobic phosphorus uptake online analyzer at full-scale was demonstrated as an effective tool to indirectly monitor the health of the PAO population and provide continuous data for real time process optimization. Understanding the conditions that improve EBPR at full-scale is important to achieve more stringent phosphorus limits that are anticipated in the future. Implementing the above strategies can reduce aeration energy consumption, metal salt and external carbon requirements, and environmental footprints at WRRFs. / Master of Science / Excess amounts of phosphorus discharged into aquatic ecosystems can lead to eutrophication, a process where algal blooms deplete the oxygen needed for other aquatic life, resulting in large scale mortalities. Water Resource Recovery Facilities (WRRFs) employ chemical, biological and/or physical means to remove phosphorus from discharged water to prevent eutrophication. Enhanced biological phosphorus removal (EBPR) is a process that takes advantage of microorganisms known as polyphosphate-accumulating organisms (PAOs) to remove phosphorus from the water. PAOs can store high amounts of phosphorus in their biomass when they are subjected to alternating anaerobic and aerobic conditions. The phosphorus is then removed with the PAOs by settling and wasting the biomass. The conventional operational approach at WRRFs is to maintain an aerobic phase with dissolved oxygen (DO) concentrations above 2 mg O2/L to maximize phosphorus uptake. However, Hampton Roads Sanitation District's Virginia Initiative Plant (VIP) observed a 69 percent decrease in average DO concentrations, alongside a 53 percent improvement in effluent phosphorus quality from 2019 to 2023. Batch tests conducted from 2020 to 2024 indicated higher phosphorus release and aerobic uptake rates at lower DO concentrations, while microbial analysis revealed a community shift toward PAOs. Recent batch tests conducted in 2023 indicated phosphorus uptake rates in the anoxic phase that ranged from 3 to 40 percent of the phosphorus uptake rates in the aerobic phase. This emergence of anoxic phosphorus uptake capacity is potentially attributable to the reduction in DO concentrations. These results highly suggest that low DO operation (DO concentrations below 1 mg O2/L) does not negatively impact EBPR performance. In fact, low DO concentrations were positively correlated with increased PAO abundance. Low DO operation can reduce aeration energy consumption, operational costs, and environmental footprints of WRRFs. Furthermore, VIP implemented other operational strategies to optimize EBPR and monitor PAO activity, including an aerobic phosphorus uptake online analyzer. This analyzer was demonstrated as an effective tool to indirectly monitor the health of the PAO population and provide continuous data for real time process optimization.

En aquest treball s'ha desenvolupat una metodologia per implementar l'eliminació biològica de fòsfor (EPBR) en les plantes de tractament d'aigües residuals urbanes (EDAR) amb configuració anòxica / òxica (A/O) dissenyades per eliminar únicament matèria orgànica (DQO) i nitrogen (N). L'objectiu és eliminar biològicament i simultàniament DQO, N i fòsfor (P) tenint en compte aspectes de control de processos i amb el millor rendiment d'operació. La metodologia proposada cerca exhaustivament un model del procés, utilitzant les dades existents de la planta per determinar els paràmetres cinètics. El model de la planta s'ha calibrat utilitzant una metodologia basada en la matriu d'informació de Fisher (FIM). Usant l'estructura del model de la planta i les noves configuracions de plantes que es proposen, s'utilitza un conjunt de criteris per identificar quina és la millor alternativa. Entre els criteris utilitzats es troben: qualitat de l'efluent, solidesa de l'estructura de control del procés, costos d'operació i costos d'inversió per a compra d'equips i per dur a terme canvis en la distribució de la planta. També s'estudia la viabilitat dels organismes acumuladors de fòsfor (PAO) i l'efecte del creixement d'aquestes espècies amb diferents estructures de control del procés. / En este trabajo se ha desarrollado una metodología para implementar la eliminación biológica de fósforo (EPBR) en las plantas de tratamiento de aguas residuales urbanas (EDAR) con configuración anóxica / óxica (A/O) diseñadas para eliminar únicamente materia orgánica (DQO) y nitrógeno (N). El objetivo es eliminar biológica y simultáneamente DQO, N y fósforo (P) teniendo en cuenta aspectos de control de procesos y con el mejor rendimiento de operación. La metodología propuesta busca exhaustivamente un modelo del proceso, utilizando los datos existentes de la planta para determinar los parámetros cinéticos. El modelo de la planta se ha calibrado utilizando una metodología basada en la matriz de información de Fisher (FIM). Usando la estructura del modelo de la planta y las nuevas configuraciones de plantas que se proponen, se utiliza un conjunto de criterios para identificar cuál es la mejor alternativa. Entre los criterios utilizados se encuentran: calidad del efluente, solidez de la estructura de control del proceso, costos de operación y costos de inversión para compra de equipos y para llevar a cabo cambios en la distribución de la planta. También se estudia la viabilidad de los organismos acumuladores de fósforo (PAO) y el efecto del crecimiento de estas especies con diferentes estructuras de control del proceso. / A methodology for retrofitting existent Anoxic/Oxic (A/O) wastewater treatment plants (WWTP) to perform the Enhanced Biological Phosphorus Removal (EPBR) in order to biologically remove organic matter (COD), nitrogen (N) and phosphorus (P) at the same time, considering process control aspects, was developed. The proposed methodology exhaustively searches a process model, using existent plant data to determine the current kinetic parameters. The plant model is calibrated using a methodology based on the Fisher Information Matrix. Using the plant model structure, new plant configurations are proposed and a set of criteria are used to identify what is the best alternative. Amongst the criteria are: the robustness of the process control structure, operating costs, investment costs to perform changes in the plant layout and equipments and the effluent quality. The feasibility of phosphorus accumulating organisms (PAO) growth and the effect of these species in the existent process control structure are also studied.

The hybrid activated sludge-biofilm system called Integrated Fixed Film Activated Sludge (IFAS) has recently become popular for enhanced nitrification and denitrification in aerobic zones because it is an alternative to increasing the volume of treatment plant units to accomplish year round nitrification and nitrogen removal. Biomass is retained on the fixed-film media and remains in the aerobic reactor, thus increasing the effective mean cell resident time (MCRT) of the biomass and providing the temperature sensitive, slow growing nitrifiers a means of staying in the system when they otherwise would washout. While the utilization of media in aerobic zones to enhance nitrification and denitrification has been the subject of several studies and full-scale experiments, the effects and performances of fixed film media integrated into the anoxic zones of biological nutrient removal (BNR) systems have not adequately been evaluated as well as the impacts of integrated media upon enhanced biological phosphorus removal (EBPR). Also, user-friendly software designed specifically to simulate the complex mixture of biological processes that occur in IFAS systems are not available. The purpose of this research was to more fully investigate the effects of integrated fixed film media on EBPR, to evaluate the impacts of media integrated into the anoxic zone on system performance, and to develop a software program that could be used to simulate the effects of integrating the various types of media into suspended growth biological nutrient removal (BNR) systems. The UCT type configuration was chosen for the BNR system, and Accuweb rope-like media was selected for integration into the anoxic zones of two IFAS systems. The media also was integrated into the aerobic reactors of one of the systems for comparison and for further investigation of the performance of the Accuweb media on enhanced nitrification and denitrification in the aerobic zones. The experiments were conducted at 10 day total MCRT during the initial phase, and then at 6 days MCRT for the experimental temperature of 10 oC. A13 hour hydraulic retention time (HRT) was used throughout the study. A high and a low COD/TP ratio were used during the investigation to further study the effects of integrated media on EBPR. The PC Windows based IFAS program began with the concepts of IAWQ model No. 2 and a zero-dimensional biofilm model was developed and added to predict the IFAS processes. Experimental data from the initial study and existing data from similar studies performed at high temperatures (>10oC) indicated that there were no significant differences in BNR performances between IFAS systems with media integrated into the anoxic and aerobic or only aerobic zones and a suspended growth control system maintained at the same relative high MCRT and temperature values. Even though greater biological nitrogen removal could not be achieved for the experimental conditions used, the experimental results indicated that the IFAS systems with fixed film media installed in the anoxic zone have a greater potential for denitrification than conventional BNR systems. As much as 30 percent of the total denitrification was observed to occur in the aerobic zones of the system installed the media only anoxic zones and 37% in the system with integrated media in both anoxic and aerobic zones where as no denitrification was observed in the aerobic zones of the control system when the systems were operated at 6 days MCRT and COD/TP of 52. It is statistically confirmed EBPR can be maintained in IFAS systems as well as Control systems, but the IFAS processes tend to have more phosphorus release in the anoxic zones with integrated fixed film installed. Further, the combination of split flow to the anoxic zone and fixed film media in the anoxic zone resulted in the decreased EBPR performances in the IFAS system relative to the control system. / Ph. D.

Grundämnet fosfor är essentiellt för alla levande organismer men kan i överskott leda till problem med övergödning. Det finns därför höga krav på halten avloppsreningsverk släpper ut till recipienter. Idag sker stora delar av fosforreningen kemiskt genom dosering av fällningskemikalier. Det finns dock fördelar med att istället använda en biologisk metod som bygger på att reningsförhållandena premierar tillväxt av bakterier med möjlighet att ta upp mer fosfor än de behöver för sin cellväxt.  Bakterierna gynnas genom omväxlande anaeroba och aeroba zoner samt en god tillgång på lättillgänglig kolkälla och fosfor. Många reningsverk kombinerar den kemiska och biologiska fosforreningen men de är inte alltid kompatibla och den kemiska kan störa ut den biologiska. På Kungsängsverket finns sedan 2010 förutsättningar för en biologisk fosforreduktion men processen har inte fungerat tillfredsställande. Anledningen tros vara höga halter järn i slammet. Järnet fäller delar av den fosfor som är nödvändig för processen. Arbetet har därför syftat till att undersöka om det går att tvätta bioslammet på järn och på så sätt nå en fungerande fosforrening; vid vilka järnhalter detta sker och vilka besparingar det skulle kunna leda till för Uppsala Vatten och Avfall AB. För vidare utredning genomfördes ett pilotförsök där två reaktorer byggdes, en referensreaktor och en försöksreaktor. Reaktorerna matades sedan med vatten med olika sammansättning, främst gällande järnhalt. Även befintlig data för verket och uppgifter kring förutsättningarna på andra reningsverk med en fungerande biologisk fosforreduktion undersöktes. Pilotförsöket visade att det går att tvätta bioslammet på järn då en sjunkande halt sågs under försökets gång. Halten sjönk från 40 mg Fe/g TS till 18 mg Fe/g TS i försöksreaktorn. En fungerande fosforrening uppnåddes aldrig så inga slutsatser gällande besparingar, eller vid vilka järnhalter en fungerande rening sker, kan dras. Andra reningsverk med biologisk fosforrening har kring 10 mg Fe/g TS vilket ger en indikation på vad halten bör vara. Pilotförsöket visade också att dosering av polymer ledde till att stora delar av den inkommande kolkällan fälldes, kolkälla som behövs för fungerande fosfor- och kväverening. Recirkulation av nitratkväve sågs hämma det fosforsläpp som vid fungerande rening ska ske i den anaeroba zonen och tros ha stört reningen under försökets gång. Förutsättningarna för biologisk fosforrening på Kungsängsvrket anses inte vara optimala gällande avloppsvattnets sammansättning, recirkulering av nitratkväve till den anaeroba zonen och mängden lättillgänglig kolkälla från hydrolysbassängen. / Phosphorus is an essential element but can cause eutrophication when present in high concentrations. Emission requirements from municipal wastewater treatment plants are therefore strict. Today chemical precipitation is common but there are advantages to using a biological method. It is based on creating conditions that favor growth of a special type of bacteria. These bacteria absorb more phosphorus than they need for growth. To do this they need alternating anaerobic and aerobic zones and access to carbon and phosphorus. A combination between the two methods are common but the precipitation chemicals can under some conditions disturb the biological removal.   At Kungsängsverket the process of biological phosphorus removal has been in place since 2010. It has not worked adequately and the reason could be high concentrations of iron in the biological sludge. The purpose of this thesis has therefore been to investigate whether it is possible to wash out the iron from the bio-sludge and as a result reach a satisfying reduction of phosphorus, to see at which iron content this might happen and what kind of savings a functioning biological phosphorus removal might lead to for Uppsala Vatten och Avfall AB.  To test the hypothesis two reactors were built, a reference reactor and an experimental reactor. The two were fed with water with different compositions, primarily regarding iron content. Also, existing data was examined from the plant and records regarding sludge composition at plants with working biological phosphorus removal. The pilot test showed that it was possible to wash out the iron from the biological sludge. Iron content in the experimental reactor went down from 40 mg Fe/g DM to 18 mg Fe/g DM. A satisfying reduction of phosphorus was never achieved and no conclusions can be drawn regarding savings or at which iron content a reduction might happen. Other wastewater treatment plants with biological phosphorus reduction have shown to have a content of about 10 mg Fe/g DM which can be used as an indication. According to the pilot test dosing of polymer can lead to a large precipitation of carbon source. Lack of carbon will inhibit phosphorus and nitrogen removal. Circulation of nitrate repressed the release of phosphate in the anaerobic zone and is believed to have disturbed the removal during the pilot. The conditions for biological phosphorus removal at Kungsängsverket are not ideal as to the composition of the wastewater, the circulation of nitrate to the anaerobic zone and the amount of carbon source from the hydrolysis.

Polyphosphat-speichernde Bakterien entfernen das im Abwasser enthaltene Phosphat durch Speicherung in Form von Granula, die dann mit einem Teil des Belebtschlammes aus dem Abwasser entfernt werden können. Dies ist wichtig um negative Einflüsse auf Oberflächengewässer wie Flüsse und Seen so gering wie möglich zu halten. Trotz intensiver Forschung ist der Prozess der sogenannten biologischen Phosphatelimination oft uneffektiv und im Jahresverlauf instabil, da über die im Belebtschlamm aktiven Polyphosphat-speichernden Bakterien nur wenig bekannt ist. Hauptproblem ist hierbei die geringe Kultivierbarkeit der Bakterien unter definierten Bedingungen (nur etwa 10-15 % der Mikroorganismen im Belebtschlamm sind kultivierbar). Aus diesem Grund war das Ziel der Arbeit die aktiven, Polyphosphat-speichernden Bakterien durchflusszytometrisch zu bestimmen und deren Dynamiken im Belebtschlamm kultivierungsunabhängig zu messen. Zunächst wurde ein Fixierungsprotokoll für die durchflusszytometrische Untersuchung der Polyphosphat-speichernden Bakterien erarbeitet, welches die größtmögliche Stabilität der hochdiversen mikrobiellen Gemeinschaft in Belebtschlammproben gewährleistet. Eine Mischung aus den Metallen Barium und Nickel (jeweils 5 mM) in einer 10%igen Natriumazidlösung erwies sich als bestes Fixierungsmittel mit einer Belebtschlamm-Stabilität von mindestens 9 Tagen. Um sowohl den DNA-als auch den Polyphosphat-Gehalt der Zellen messen zu können wurde weiterhin eine neue und sehr spezifische Polyphosphatfärbung auf Basis des fluoreszierenden Antibiotikums Tetrazyklin etabliert. Tetrazyklin bindet divalente Kationen, die auch in großer Menge in Polyphosphatgranula enthalten sind und fluoresziert gelblich grün. Die entwickelten Methoden zur Fixierung und Polyphosphatfärbung wurden an Belebtschlamm einer kommunalen Kläranlage getestet. Neben DNA- und Polyphosphat-Gehalt der Bakterienzellen wurde eine Vielzahl abiotischer Parameter (pH, Temperatur, Leitfähigkeit, …) gemessen. Diese wurden zusammen mit den durchflusszytometrischen Daten mittels Korrelationsanalyse ausgewertet. Hieraus ergaben sich wichtige Hinweise auf die Art der Polyphosphat-speichernden Bakterien, fördernde und störende Einflüsse des in der Kläranalage behandelten Abwassers auf die biologische Phosphatelimination und die Abhängigkeiten der mikrobiellen Gemeinschaft von Faktoren wie Temperatur, pH oder der anfallenden Regenmenge. Diese Erkenntnisse können genutzt werden um die biologische Phosphatelimination aus dem Abwasser zu verbessern und damit den Weg zu einer Ressourcen- und Umweltschonenden Phosphatrückgewinnung zu bereiten. Außerdem ist es, bei Kenntnis des kläranlagenspezifischen Prozesses, möglich anhand der durchflusszytometrischen Daten schnell die aktuelle Situation zu erfassen und gegebenenfalls rechtzeitig auf Änderungen zu reagieren, bevor es zu einer massiven Störung kommt. Eine Kombination von Durchflusszytometrie und der Erfassung abiotischer Daten ist nicht nur auf die biologische Phosphateliminierung anwendbar, sondern auch auf viele andere wissenschaftliche Fragestellungen.

Enhanced Biological Phosphorus Removal (EBPR) is an established process in wastewater treatment that uses bacteria to reduce phosphorus levels below regulatory discharge limits. Recently, in light of growing political concern over phosphorus sustainability, EBPR has also been recognised as a platform from which phosphorus may be recovered and recycled onto land as fertiliser. Operating EBPR to optimise performance and efficiency is therefore extremely important, but remains a challenge due to poor understanding of these bacterial ecosystems. Raman spectroscopy is a non-invasive, label-free, culture-independent technique capable of analysing live, single cells. Despite its advantages, Raman spectroscopy has been applied to study EBPR bacteria in just a handful of studies and thus has a low profile in this field of research. More work is required to investigate potential areas of application for Raman spectroscopy in EBPR research. The principal thesis presented here is that Raman spectroscopy could be used as a tool to improve EBPR. The Raman spectra used for this investigation were acquired from individual EBPR bacteria dried onto a calcium fluoride substrate. The bacterial samples were collected from three different sources, namely lab-scale sequencing batch reactors located in Edinburgh (University of Edinburgh, UK) and Boston (Northeastern University, USA), and a full-scale EBPR plant in Slough (Thames Water, UK). Using these spectra, some potential applications and limitations of Raman spectroscopy for improving EBPR were explored. In this foundation work, a particular emphasis on spectral analysis methods was kept in light of the benefits of automating analysis as well as the need for standardisation to be able to compare results between different studies and groups. Nine methods were compared for baselining Raman spectra of individual EBPR bacteria. From these, the “small-window moving average” (SWiMA) method was determined to be the best baselining technique for our purposes at the current time. In agreement with earlier studies, the Raman spectroscopic signatures of three key EBPR metabolites – polyphosphate, polyhydroxyalkanoate (PHA) and glycogen – were shown to be clearly identifiable in individual EBPR bacteria when present. The Raman shifts of characteristic spectral bands arising from polyphosphate were shown to vary significantly between samples and the implications of this were discussed. Examples of how the Raman spectra of individual bacteria can be modelled with multivariate tools to open up new areas for research were given. MCR modelling was demonstrated to offer a novel way to normalise the Raman spectra of individual EBPR bacteria prior to quantitative analysis. With the instrumental set-up in this work, the limit of detection (LOD) of aqueous polyphosphate samples was estimated to be approximately 0.08 M and 0.02 M for 10 second and 200 second acquisitions respectively. Future work is required to research ways in which a more comparable form of polyphosphate ‘standard’ might be prepared so that direct correlation can be drawn between measurements made on such a standard and measurements made in bacterial cells. Overall, several applications and challenges of Raman spectroscopy for the investigation of EBPR bacteria are presented in this work together with recommendation for how to process the spectral data. The conclusions drawn from this work indicate that Raman spectroscopy could be used as a tool to improve EBPR but further work is required to refine and apply these methods.

Submitted by Fernanda Rodrigues de Lima (fernanda.rlima@ufpe.br) on 2018-07-23T21:48:00Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇAO Cândida Nathaly Cordeiro Souto.pdf: 2081125 bytes, checksum: e4bcbb31239d576354a7bab1aa383a1c (MD5) / Approved for entry into archive by Alice Araujo (alice.caraujo@ufpe.br) on 2018-07-24T18:34:04Z (GMT) No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇAO Cândida Nathaly Cordeiro Souto.pdf: 2081125 bytes, checksum: e4bcbb31239d576354a7bab1aa383a1c (MD5) / Made available in DSpace on 2018-07-24T18:34:04Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) DISSERTAÇAO Cândida Nathaly Cordeiro Souto.pdf: 2081125 bytes, checksum: e4bcbb31239d576354a7bab1aa383a1c (MD5) Previous issue date: 2017-02-20 / FACEPE / A disponibilidade hídrica é uma das maiores preocupações ambientais a nível mundial. O aumento da demanda por água sugere uma necessidade maior do tratamento e reutilização das águas consumidas. A eutrofização dos corpos hídricos é um problema ambiental recorrente, causado pelas descargas excessivas de nutrientes nos corpos hídricos. Os reatores em bateladas sequenciais (RBS) possibilitam a remoção de nutrientes em esgotos através de processos biológicos. Uma vez que a população microbiológica é exposta a condições alternadas anaeróbia/aeróbia/anóxica, o fósforo é removido devido à atividade de micro-organismos que o acumulam; simultaneamente, o nitrogênio é removido por nitrificação e desnitrificação. Neste trabalho, um RBS de 140 L de volume útil foi operado recebendo esgoto doméstico de baixa carga orgânica proveniente do tratamento preliminar de uma ETE. O estudo foi dividido em duas fases, em que foram alterados os tempos de ciclos e das fases de reação. O sistema foi monitorado por meio de coletas semanais para análise das variáveis de interesse. Quando se atingiu o estado estacionário, foram traçados perfis do funcionamento do reator em cada fase operacional. Para isto, foram realizadas análises físico-químicas (fósforo, nitrogênio, sólidos, alcalinidade, DQO, AGV e PHA) e microbiológicas (microscopia óptica, testes de atividade da biomassa e teste de biologia molecular por PCR). Os resultados das variáveis físico-químicas mostraram eficiência na remoção de fósforo, nitrogênio e matéria orgânica; O processo EBPR se efetivou conforme o esperado em ambas as fases; sugerindo a atividade de PAOs e DPAOs no sistema. A remoção de nitrogênio amoniacal foi completa e efetiva. Os testes biomoleculares confirmaram a presença no sistema operado dos principais grupos de micro-organismos responsáveis pela remoção biológica de nutrientes: PAOs, Anammox, desnitrificantes e nitrificantes. / The water availability is one of the major environmental concerns worldwide. The increased demand for water suggests a greater need for treatment and reutilization of the wastewater. The eutrophication of the water bodies is a recurrent environmental issue, caused by the excessive discharge of nutrients in the water bodies. Sequencing batch reactors (SBR) allow the removal of nutrients from sewage by biological processes. Once the microbiological population is exposed to alternate anaerobic/aerobic/anoxic conditions, the phosphorus is removed due the activity of the microorganisms that accumulate it; at the same time, the nitrogen is removed by nitrification and denitrification. In this work, one SBR with 140 L of working volume was operated receiving a low organic load of domestic sewage from the preliminary treatment of a sewage treatment plant. The study was divided in two phases, in which the times of the cycles and of the reaction phases were changed. The system was monitored by weekly sampling for analysis of the parameter of interest. When the stationary state was reached, the operating profiles of the reactor in each operational phase were plotted. For this aim, physical-chemical (phosphorus, nitrogen, solids, alkalinity, COD, VFA and PHA) and microbiological (optical microscopy, biomass activity test and molecular biology test by PCR) analysis were performed. The physical-chemical results showed efficiency on the removal of phosphorus, nitrogen and organic matter; The EBPR process took effect as expected in both phases; suggesting the activity of PAO and DPAO in the system. The removal of ammonia nitrogen was complete and effective. The biomolecular tests confirmed the presence of the main microorganisms groups responsible for the biological removal of nutrients in the operated system: PAO, Anammox, denitrifying, nitrifying.

pH control is a tool to improve some aspects of Enhanced Biological Phosphorus Removal (EBPR) process. Filipe et al (2001a, 2001b, and 2001c) found strong evidence that the stability of EBPR systems can be improved by increasing the pH of the anaerobic zone, thereby creating conditions where phosphorus-accumulating organisms (PAOs) are able to take up acetate faster than glycogen-accumulating organisms (GAOs). They explained this observation by comparing the growth rate of phosphorus-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs) and found that pH has little effect on PAOs growth rate but adversely affects GAOs growth rate when it increases (at pH values greater than 7.25, PAOs would take acetate faster than GAOs would). They used synthetic wastewater rich in acetic acid. In this study, we used real wastewater and the dominant volatile fatty acid available to microorganisms was propionic acid in continuous EBPR system. It was found that lower anaerobic zone pH (6.5 vs. 7.2) reduced the anaerobic P release both on an MLVSS specific basis and also on a non-specific (absolute value for the process) basis. In addition, the observed yield was significantly decreased. Aerobic P uptake was lower in the low-pH system (on a non-specific basis) due to the lower observed yield, and thus lower MLVSS concentration. Net P uptake was hard to interpret because of the effect of P release in the secondary clarifier of Train 2 (high pH). However, on a specific basis it was clear that net P uptake was either equal or better in the low-pH system regardless of how the secondary clarifier data was interpreted. Carbon transformations were not impacted in as consistent a fashion as anaerobic P release was. On a specific basis, PHA content remained unchanged although the PHV/PHB ratio was impacted with much lower PHV content in the low-pH system. Glycogen content and the amount of labile glycogen (delta glycogen) were higher in the low-pH system, in spite of the fact that MLVSS P content did not decrease. However, due to the impact of the low observed yield at low pH, absolute values resulted in higher PHA content for the process reactors as a whole, higher glycogen content, and unchanged labile glycogen. Low pH resulted in increased biomass P content, however the lower observed yield offset this on a process basis so that effluent P levels were nearly equal. So low pH improved P removal on a specific basis, but not on a process basis. Since it is unknown if the low observed yield is repeatable, and due to the impact of the secondary clarifier in the high pH system, it cannot be concluded that the effect of low pH on net P removal would be similar in other EBPR systems.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-07-26T14:51:03Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Ferreira, Ana Linda (TESE) EngCivil14.pdf: 3085122 bytes, checksum: d948f8fbcdf65b938ccf579d975895d1 (MD5) / Made available in DSpace on 2016-07-26T14:51:03Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Ferreira, Ana Linda (TESE) EngCivil14.pdf: 3085122 bytes, checksum: d948f8fbcdf65b938ccf579d975895d1 (MD5) Previous issue date: 2014-06-30 / CAPEs / O aporte excessivo de nutrientes como o fósforo e o nitrogênio são uma das principais causas da eutrofização das águas, um grave problema ambiental resultante de descargas de efluentes domésticos e industriais, bem como do uso intensivo de fertilizantes em campos agrícolas. Não obstante, é preocupante o facto das reservas de fósforo estarem a diminuir mundialmente. Este trabalho objetiva a remoção simultânea de fósforo e nitrogênio e o estudo das condições de funcionamento para o estabelecimento de uma comunidade específica de microrganismos para esse fim, tratando efluente real de origem domestica. Para isso foram instalados dois reatores em batelada sequencial com 140L de volume útil na ETE Mangueira em Recife/PE. Os resultados revelam que a adição de propionato para o enriquecimento do lodo melhora a sua sedimentabilidade enquanto que a adição de acetato favorece o desenvolvimento de organismos filamentosos. A aplicação de aeração continua inibiu a nitrificação e desnitrificação simultânea, enquanto que a aeração escalonada favoreceu esse processo. Dependendo da forma de como se procede a aeração do reator é possível reduzir a fonte de carbono na fase anóxica e também o TDH global de operação do reator. Foi possível remover DQO com eficiência acima de 90%, além de remover nitrogênio e fósforo em reatores RSB, mesmo com esgotos diluídos. A remoção de fósforo ocorreu através do mecanismo de remoção biológica de fósforo avançado (EBPR). / Excessive discharges of nutrients like phosphorus and nitrogen are one of the main causes of water eutrophication, a serious environmental problem resulting mainly from discharges of domestic and industrial effluents as well as the intensive use of fertilizers in agricultural fields. Nevertheless, it is being motif of concern that phosphorus reserves are declining worldwide. This work aims for simultaneous removal of phosphorus and nitrogen and the study of the operating conditions for the establishment of a specific community of microorganisms for this purpose, treating real domestic wastewater. For this two reactors were installed in sequencing batch with 140L working volume in ETE Mangueira in Recife / PE. The results show that the addition of propionate to the enrichment of the sludge settle ability improves their while adding acetate favors the growth of filamentous organisms. The application of continued aeration inhibited simultaneous nitrification and denitrification (SND), while the stepped aeration favored this process. Depending on how one proceeds as aeration reactor is possible to reduce the carbon source in the anoxic phase and also the overall HRT of reactor operation. COD could be removed with efficiency above 90%, in addition to removing nitrogen and phosphorus in SBR reactors, even with diluted sewage. The phosphorus removal occurred through the mechanisms of Enhanced Biological Phosphorus Removal(EBPR).

Polyphosphat-speichernde Bakterien entfernen das im Abwasser enthaltene Phosphat durch Speicherung in Form von Granula, die dann mit einem Teil des Belebtschlammes aus dem Abwasser entfernt werden können. Dies ist wichtig um negative Einflüsse auf Oberflächengewässer wie Flüsse und Seen so gering wie möglich zu halten. Trotz intensiver Forschung ist der Prozess der sogenannten biologischen Phosphatelimination oft uneffektiv und im Jahresverlauf instabil, da über die im Belebtschlamm aktiven Polyphosphat-speichernden Bakterien nur wenig bekannt ist. Hauptproblem ist hierbei die geringe Kultivierbarkeit der Bakterien unter definierten Bedingungen (nur etwa 10-15 % der Mikroorganismen im Belebtschlamm sind kultivierbar). Aus diesem Grund war das Ziel der Arbeit die aktiven, Polyphosphat-speichernden Bakterien durchflusszytometrisch zu bestimmen und deren Dynamiken im Belebtschlamm kultivierungsunabhängig zu messen. Zunächst wurde ein Fixierungsprotokoll für die durchflusszytometrische Untersuchung der Polyphosphat-speichernden Bakterien erarbeitet, welches die größtmögliche Stabilität der hochdiversen mikrobiellen Gemeinschaft in Belebtschlammproben gewährleistet. Eine Mischung aus den Metallen Barium und Nickel (jeweils 5 mM) in einer 10%igen Natriumazidlösung erwies sich als bestes Fixierungsmittel mit einer Belebtschlamm-Stabilität von mindestens 9 Tagen. Um sowohl den DNA-als auch den Polyphosphat-Gehalt der Zellen messen zu können wurde weiterhin eine neue und sehr spezifische Polyphosphatfärbung auf Basis des fluoreszierenden Antibiotikums Tetrazyklin etabliert. Tetrazyklin bindet divalente Kationen, die auch in großer Menge in Polyphosphatgranula enthalten sind und fluoresziert gelblich grün. Die entwickelten Methoden zur Fixierung und Polyphosphatfärbung wurden an Belebtschlamm einer kommunalen Kläranlage getestet. Neben DNA- und Polyphosphat-Gehalt der Bakterienzellen wurde eine Vielzahl abiotischer Parameter (pH, Temperatur, Leitfähigkeit, …) gemessen. Diese wurden zusammen mit den durchflusszytometrischen Daten mittels Korrelationsanalyse ausgewertet. Hieraus ergaben sich wichtige Hinweise auf die Art der Polyphosphat-speichernden Bakterien, fördernde und störende Einflüsse des in der Kläranalage behandelten Abwassers auf die biologische Phosphatelimination und die Abhängigkeiten der mikrobiellen Gemeinschaft von Faktoren wie Temperatur, pH oder der anfallenden Regenmenge. Diese Erkenntnisse können genutzt werden um die biologische Phosphatelimination aus dem Abwasser zu verbessern und damit den Weg zu einer Ressourcen- und Umweltschonenden Phosphatrückgewinnung zu bereiten. Außerdem ist es, bei Kenntnis des kläranlagenspezifischen Prozesses, möglich anhand der durchflusszytometrischen Daten schnell die aktuelle Situation zu erfassen und gegebenenfalls rechtzeitig auf Änderungen zu reagieren, bevor es zu einer massiven Störung kommt. Eine Kombination von Durchflusszytometrie und der Erfassung abiotischer Daten ist nicht nur auf die biologische Phosphateliminierung anwendbar, sondern auch auf viele andere wissenschaftliche Fragestellungen.