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Low dissolved oxygen (DO) biological nutrient removal (BNR) is becoming a viable option to improve the energy efficiency of BNR. To properly model and design BNR processes for low DO operation, it is critical to fully understand how nitrifier, denitrifier, and polyphosphate accumulating organism (PAO) oxygen kinetics adapt in a shift from traditional DO operation (2 mg O2/L or more) to low DO operation. Research characterizing how oxygen kinetics shift over time in activated sludge biomass adapting to low DO is limited. Therefore, a method to characterize oxygen kinetics for nitrifiers, denitrifiers, and PAOs simultaneously is lacking. Here a method was developed to simultaneously measure the oxygen kinetics of nitrifiers, denitrifiers, and PAOs. This method, termed the SND and P-Uptake Oxygen Kinetics test, was able to estimate the ammonia oxidizing bacteria (AOB) oxygen half-saturation coefficient, ammonia maximum removal rate, denitrifier oxygen inhibition coefficient, total inorganic nitrogen (TIN) maximum removal rate, PAO oxygen half-saturation coefficient, phosphorus maximum uptake rate, and a simultaneous nitrification and denitrification (SND) optimum operation point. Three tests were conducted on the Virginia Initiative Plant (VIP) BNR Activated Sludge Pilot while it was operating at a process DO of 2 mg O2/L, and one test while it was operating at 1.5 mg O2/L. The measurements among the three initial tests showed high similarity in their parameter estimates. Estimated oxygen half-saturation and oxygen inhibition coefficients were compared to current suggested ranges and were within the expected magnitudes. At 2 mg O2/L, denitrifier oxygen inhibition coefficients and PAO oxygen half-saturation coefficients were estimated to be remarkably low here, under 0.4 and 0.1 mg O2/L, respectively. AOB oxygen half-saturation coefficients were variable here in the range of 0.62 to 2.57 mg O2/L, seeming to vary with available ammonia concentrations. Upon comparison wit

Sialic acids are a family of nine-carbon negatively charged carbohydrates. In animals, they are abundant on mucosa surfaces as terminal carbohydrates of mucin glycoproteins. Some commensal and pathogenic bacteria are able to release, take up and catabolize sialic acids. Recently, sialic acids have been discovered to be widespread among most microorganisms. Although the catabolism of sialic acids has been intensively investigated in the field of host-microbe interactions, very limited information is available on microbial degradation of sialic acids produced by environmental microorganisms. In this study, the catabolic pathways of sialic acids within a microbial community dominated by 'Candidatus Accumulibacter' were evaluated. Protein alignment tools were used to detect the presence of the different proteins involved in the utilization of sialic acids in the flanking populations detected by 16S rRNA gene amplicon sequencing. The results showed the ability of Clostridium to release sialic acids from the glycan chains by the action of a sialidase. Clostridium and Chryseobacterium can take up free sialic acids and utilize them as nutrient. Interestingly, these results display similarities with the catabolism of sialic acids by the gut microbiota. This study points at the importance of sialic acids in environmental communities in the absence of eukaryotic hosts., Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public., BT/Environmental Biotechnology, Environmental Fluid Mechanics

Sialic acids are a family of nine-carbon negatively charged carbohydrates. In animals, they are abundant on mucosa surfaces as terminal carbohydrates of mucin glycoproteins. Some commensal and pathogenic bacteria are able to release, take up and catabolize sialic acids. Recently, sialic acids have been discovered to be widespread among most microorganisms. Although the catabolism of sialic acids has been intensively investigated in the field of host–microbe interactions, very limited information is available on microbial degradation of sialic acids produced by environmental microorganisms. In this study, the catabolic pathways of sialic acids within a microbial community dominated by ‘Candidatus Accumulibacter’ were evaluated. Protein alignment tools were used to detect the presence of the different proteins involved in the utilization of sialic acids in the flanking populations detected by 16S rRNA gene amplicon sequencing. The results showed the ability of Clostridium to release sialic acids from the glycan chains by the action of a sialidase. Clostridium and Chryseobacterium can take up free sialic acids and utilize them as nutrient. Interestingly, these results display similarities with the catabolism of sialic acids by the gut microbiota. This study points at the importance of sialic acids in environmental communities in the absence of eukaryotic hosts.

Tetrasphaera polyphosphate accumulating organisms (PAOs) have been found in greater abundance and may remove more P in full-scale enhanced biological phosphorus removal (EBPR) water resource recovery facilities (WRRF) worldwide than the more extensively studied Accumulibacter PAO. Understanding the effect that environmental conditions, such as pH, have on Tetrasphaera metabolisms is critical to understanding EBPR WRRF process upsets, achieving consistent low effluent P concentrations, and formulating optimization strategies. The objective of this study was to determine pH effects over the range from 5.5 to 8.5 on anaerobic amino acid uptake, P release, and cation release in a Tetrasphaera-enriched culture from a lab-scale sequencing batch reactor. Lower rates of carbon uptake and coupled P release were found in this study than in similar studies of Accumulibacter. Extracellular pH during carbon uptake was correlated to the change in pH, P release, and cation release, whereas carbon uptake and NH4-N release were not correlated to pH. The ratio of P released to carbon taken up was like other studies of Accumulibacter suggesting similar P release driven secondary transport mechanisms may be used by Tetrasphaera. The ability of Tetrasphaera PAOs to take up organic carbon over a wide range of pH conditions reinforces the idea that they may be more metabolically heterogeneous and adaptable to varied conditions than Accumulibacter. Optimization of EBPR WRRFs may benefit from strategies that support both a robust Accumulibacter and Tetrasphaera community.

Tetrasphaera polyphosphate accumulating organisms (PAOs) have been found in greater abundance and may remove more P in full-scale enhanced biological phosphorus removal (EBPR) water resource recovery facilities (WRRF) worldwide than the more extensively studied Accumulibacter PAO. Understanding the effect that environmental conditions, such as pH, have on Tetrasphaera metabolisms is critical to understanding EBPR WRRF process upsets, achieving consistent low effluent P concentrations, and formulating optimization strategies. The objective of this study was to determine pH effects over the range from 5.5 to 8.5 on anaerobic amino acid uptake, P release, and cation release in a Tetrasphaera-enriched culture from a lab-scale sequencing batch reactor. Lower rates of carbon uptake and coupled P release were found in this study than in similar studies of Accumulibacter. Extracellular pH during carbon uptake was correlated to the change in pH, P release, and cation release, whereas carbon uptake and NH4-N release were not correlated to pH. The ratio of P released to carbon taken up was like other studies of Accumulibacter suggesting similar P release driven secondary transport mechanisms may be used by Tetrasphaera. The ability of Tetrasphaera PAOs to take up organic carbon over a wide range of pH conditions reinforces the idea that they may be more metabolically heterogeneous and adaptable to varied conditions than Accumulibacter. Optimization of EBPR WRRFs may benefit from strategies that support both a robust Accumulibacter and Tetrasphaera community.

Seawater can be introduced or intrude in sewer systems and can thereby negatively influence biological wastewater treatment processes. Here we studied the impact of artificial seawater on the enhanced biological phosphate removal (EBPR) process performance by aerobic granular sludge (AGS) with synthetic wastewater. Process performance, granule stability and characteristics as well as microbial community of a seawater-adapted AGS system were observed. In seawater conditions strong and stable granules formed with an SVI5 of 20 mL/g and a lower abrasion coefficient than freshwater-adapted granules. Complete anaerobic uptake of acetate, anaerobic phosphate release of 59.5 ± 4.0 mg/L PO43--P (0.35 mg P/mg HAc), and an aerobic P-uptake rate of 3.1 ± 0.2 mg P/g VSS/h were achieved. The dominant phosphate accumulating organisms (PAO) were the same as for freshwater-based aerobic granular sludge systems with a very high enrichment of Ca. Accumulibacter phosphatis clade I, and complete absence of glycogen accumulating organisms. The effect of osmotic downshocks was tested by replacing influent seawater-based medium by demineralized water-based medium. A temporary decrease of the salinity in the reactor led to a decreased phosphate removal activity, while it also induced a rapid release of COD by the sludge, up to 45.5 ± 1.7 mg COD/g VSS. This is most likely attributed to the release of osmolytes by the cells. Recovery of activity was immediately after restoring the seawater feeding. This work shows that functioning of aerobic granular sludge in seawater conditions is as stable as in freshwater conditions, while past research has shown a negative effect on operation of AGS processes with NaCl-based wastewater at the same salinity as seawater., BT/Environmental Biotechnology

Seawater can be introduced or intrude in sewer systems and can thereby negatively influence biological wastewater treatment processes. Here we studied the impact of artificial seawater on the enhanced biological phosphate removal (EBPR) process performance by aerobic granular sludge (AGS) with synthetic wastewater. Process performance, granule stability and characteristics as well as microbial community of a seawater-adapted AGS system were observed. In seawater conditions strong and stable granules formed with an SVI5 of 20 mL/g and a lower abrasion coefficient than freshwater-adapted granules. Complete anaerobic uptake of acetate, anaerobic phosphate release of 59.5 ± 4.0 mg/L PO43--P (0.35 mg P/mg HAc), and an aerobic P-uptake rate of 3.1 ± 0.2 mg P/g VSS/h were achieved. The dominant phosphate accumulating organisms (PAO) were the same as for freshwater-based aerobic granular sludge systems with a very high enrichment of Ca. Accumulibacter phosphatis clade I, and complete absence of glycogen accumulating organisms. The effect of osmotic downshocks was tested by replacing influent seawater-based medium by demineralized water-based medium. A temporary decrease of the salinity in the reactor led to a decreased phosphate removal activity, while it also induced a rapid release of COD by the sludge, up to 45.5 ± 1.7 mg COD/g VSS. This is most likely attributed to the release of osmolytes by the cells. Recovery of activity was immediately after restoring the seawater feeding. This work shows that functioning of aerobic granular sludge in seawater conditions is as stable as in freshwater conditions, while past research has shown a negative effect on operation of AGS processes with NaCl-based wastewater at the same salinity as seawater., BT/Environmental Biotechnology

Control of wastewater treatment plants has received more attention as a method to improve nutrient removal processes. Improved nutrient removal, results in an increased effluent quality, whilst minimising the energy consumption. Aeration control of oxidation ditches often focuses on adjusting the aeration intensity based on the nitrogen removal process, using ammonium measurements. The control systems do not incorporate biological phosphorus removal. Biological phosphorus removal, on the one hand, needs sufficient aeration and on the other hand, can deteriorate when subjected to excessive aeration. The complex relation to aeration, makes control of biological phosphorus removal difficult. This research investigates the control of biological phosphorus removal in an oxidation ditch using simple sensors. Control systems often require expensive instrumentation. To limit the costs, cheap sensors using alternative monitoring variables for biological phosphorus removal are desired. Literature research was done to investigate the principles of biological phosphorus removal, possible alternative monitoring variables, current control systems and existing mathematical models and benchmarks that could contribute to a renewed control strategy. Some important variables found in literature were potassium, pH, ORP, ortho-P, nitrate, ammonium, conductivity, and DO. A measurement campaign was set up and executed to measure the trend of these variables throughout the oxidation ditch and to obtain more information on the state of the variables in the (un)aerated zones. A full-scale operating wastewater treatment plant in Hattem was made available to execute the measurement campaign. The results obtained from the measuring campaign did not point towards alternative monitoring variables for biological phosphorus removal. Relationships between the variables and phosphorus found in literature, could not be directly derived from the measurement results. However, nitrate and amm

Control of wastewater treatment plants has received more attention as a method to improve nutrient removal processes. Improved nutrient removal, results in an increased effluent quality, whilst minimising the energy consumption. Aeration control of oxidation ditches often focuses on adjusting the aeration intensity based on the nitrogen removal process, using ammonium measurements. The control systems do not incorporate biological phosphorus removal. Biological phosphorus removal, on the one hand, needs sufficient aeration and on the other hand, can deteriorate when subjected to excessive aeration. The complex relation to aeration, makes control of biological phosphorus removal difficult. This research investigates the control of biological phosphorus removal in an oxidation ditch using simple sensors. Control systems often require expensive instrumentation. To limit the costs, cheap sensors using alternative monitoring variables for biological phosphorus removal are desired. Literature research was done to investigate the principles of biological phosphorus removal, possible alternative monitoring variables, current control systems and existing mathematical models and benchmarks that could contribute to a renewed control strategy. Some important variables found in literature were potassium, pH, ORP, ortho-P, nitrate, ammonium, conductivity, and DO. A measurement campaign was set up and executed to measure the trend of these variables throughout the oxidation ditch and to obtain more information on the state of the variables in the (un)aerated zones. A full-scale operating wastewater treatment plant in Hattem was made available to execute the measurement campaign. The results obtained from the measuring campaign did not point towards alternative monitoring variables for biological phosphorus removal. Relationships between the variables and phosphorus found in literature, could not be directly derived from the measurement results. However, nitrate and amm

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, 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

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, 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

This project utilizes hydrocyclones in wastewater treatment to select for heavier solids, and has been used before in multiple small-scale systems. This is the first implementation of hydrocyclones in a full-scale plant for the purpose of increased settleability, while also achieving enhanced biological phosphorus removal without the use of an anaerobic selector. Hydrocyclones receive mixed liquor tangentially and separate light solids from more dense solids through their tapered shape, increasing the velocity of liquid as it moves downward and allowing for selection of a certain solids fraction. The hydrocyclones receive flow from the waste stream, selecting for dense solids to recycle through the process while light solids are wasted, creating a balance of granules and flocs with superior settling characteristics in which phosphorus is removed through phosphorus accumulating organisms (PAO). This project was implemented at a wastewater treatment plant rated at 20 MGD utilizing a 4-stage Bardenpho configuration with an IFAS system. This plant routinely experienced moderate settleability issues with an average SVI of 141 and a 90th percentile SVI of 179. Over time data was collected to characterize settleability and activity of PAO, GAO, and filaments. Using an external selector to achieve biological phosphorus is significant in that most wastewater treatment plants cannot do this without the use of an anaerobic selector. This has the potential to apply external selectors to existing infrastructure throughout plants worldwide to achieve not only biological phosphorus removal, but also improved settleability with a very minor capital investment.

The success of enhanced biological phosphorus removal depends on the constant availability of volatile fatty acids (VFAs). To reduce costs of purchasing external carbon, waste streams would be a preferred source for nutrient removal. VFAs were shown to vary in the incoming sewage and fermentate from primary sludge (PS). Another available source of organic to generate VFAs is waste activated sludge (WAS). The effect of solids retention time and biomass concentration, as well as the effect of temperature and requirement for mixing on generation of VFA from the fermentation of WAS were investigated. It was found that VFA yields from sludge fermentation increased with SRT. At the longest SRT of 10 days improved biomass degradation resulted in the highest soluble to total COD ratio and the highest VFA yield. WAS fermentation was found highly temperature-dependent. The overall VFA–COD concentration in the non-mixed reactors was much lower than the mixed reactors. The study of fermentation of PS, WAS and a mixture of WAS and PS demonstrated that PS fermentation predictably generated a significantly higher amount of soluble COD than WAS. Co-fermentation of WAS with PS enhanced soluble COD production and increased the release of phosphate and ammonium. Fermentation of combined PS and WAS sludge generated a concentration of phosphate high enough to allow phosphorus recovery as struvite The effect of using glycerol as an external carbon source in biological phosphorus removal was investigated. Using glycerol directly resulted in the failure of the process which maintained enhanced biological phosphorus removal. When glycerol was co-fermented with waste activated sludge, significant VFA production was observed. By 2 | P a g e supplying the system with the VFA-enriched supernatant of the fermentate, biological phosphorus removal was enhanced. It was concluded that, if glycerol was to be used as external carbon source for biological phosphorous removal, the effective approach was

The success of enhanced biological phosphorus removal depends on the constant availability of volatile fatty acids (VFAs). To reduce costs of purchasing external carbon, waste streams would be a preferred source for nutrient removal. VFAs were shown to vary in the incoming sewage and fermentate from primary sludge (PS). Another available source of organic to generate VFAs is waste activated sludge (WAS). The effect of solids retention time and biomass concentration, as well as the effect of temperature and requirement for mixing on generation of VFA from the fermentation of WAS were investigated. It was found that VFA yields from sludge fermentation increased with SRT. At the longest SRT of 10 days improved biomass degradation resulted in the highest soluble to total COD ratio and the highest VFA yield. WAS fermentation was found highly temperature-dependent. The overall VFA–COD concentration in the non-mixed reactors was much lower than the mixed reactors. The study of fermentation of PS, WAS and a mixture of WAS and PS demonstrated that PS fermentation predictably generated a significantly higher amount of soluble COD than WAS. Co-fermentation of WAS with PS enhanced soluble COD production and increased the release of phosphate and ammonium. Fermentation of combined PS and WAS sludge generated a concentration of phosphate high enough to allow phosphorus recovery as struvite The effect of using glycerol as an external carbon source in biological phosphorus removal was investigated. Using glycerol directly resulted in the failure of the process which maintained enhanced biological phosphorus removal. When glycerol was co-fermented with waste activated sludge, significant VFA production was observed. By 2 | P a g e supplying the system with the VFA-enriched supernatant of the fermentate, biological phosphorus removal was enhanced. It was concluded that, if glycerol was to be used as external carbon source for biological phosphorous removal, the effective approach was

Toxic shock events can be detrimental to wastewater treatment systems and can result in long-term losses of system performance. If warned of an impending toxic shock, operators would have the opportunity to implement process controls that could help mitigate the effects of the shock event. The objective of this project was to evaluate the effectiveness of a developed corrective action strategy (involving aerobic endogenous respiration) on an enhanced biological phosphorus removal (EBPR) wastewater treatment plant (WWTP) shocked with chlorine. Three identical, laboratory-scale systems were designed to mimic one train of the Long Creek Water Resources Reclamation Facility (WRRF, Gastonia, NC). The basis of this study is a comparative performance analysis among the three trains; a negative control (unshocked and operated normally), a positive control (shocked with hypochlorite and operated normally), and the corrective action (shocked with hypochlorite and process controls implemented). Comparative performance analysis among the three trains was based on effluent quality, performance stability, and biomass kinetics as indicated by rates of respiration and phosphate release and uptake. The shock event and corrective action strategy both inhibited EBPR. After an initial perturbation, the positive control matched the performance of the negative control. The corrective action, however, exhibited significant instability in EBPR performance. Regardless of whether aerobic or anaerobic sludge storage conditions are selected, endogenous respiration will still result in system instability. It is recommended, therefore, that measures be taken to avoid imposing endogenous conditions on isolated sludge during a short-term toxic shock event.

Waste Stabilisation Ponds (WSPs) are characterised by low phosphorus (P) removal capacity. Heterotrophic bacteria are principal microbial agents in WSPs in addition to algae. As treatment proceeds in WSPs, algal growth increases and pH rises, this has lead to believe that P removal is mainly through sedimentation as organic P algal biomass and precipitation as inorganic P. In activated sludge treatment plants (AS), microbial P removal has been improved and is termed as enhanced biological phosphorus removal. There was a need to establish whether it was possible to enhance P removal in WSPs. A performance assessment of pond system at the University of Dare s Salaam (UDSM), Tanzania, has shown that 90% of the P removed was in the primary pond (facultative) and the rest in the maturation pond (aerobic). In these studies, a pure strain A. hydrophyla was isolated from an activated sludge wastewater treatment plant in Sweden. This plant has a train that functions with enhanced biological phosphorus removal. The strain was tested for P uptake in minimal media supplemented with glucose, succinate or acetate, grown aerobically and anaerobically/aerobically. This strain was able to take up P without having been subjected to the anaerobic phase. It was observed that P uptake was enhanced after the anaerobic phase with media supplemented with glucose, but not with succinate or acetate. Phosphorus uptake repeatedly followed the bacterial growth pattern with correlation coefficients of more than 95%. Therefore P removal has a direct correlation with bacterial growth. Two isolates Acinetobacter sp. (isolated from the primary facultative pond) and E .coli (isolated from the maturation pond) were obtained from a tropical WSP treatment system at the UDSM. They were subjected to aerobic P uptake experiment similar to those of A.hydrophyla. The uptake per unit absorbance of bacterial growth was found to be comparable to that of A.hydrophyla, isolated from AS. These results showed that, QC 20101119

Waste Stabilisation Ponds (WSPs) are characterised by low phosphorus (P) removal capacity. Heterotrophic bacteria are principal microbial agents in WSPs in addition to algae. As treatment proceeds in WSPs, algal growth increases and pH rises, this has lead to believe that P removal is mainly through sedimentation as organic P algal biomass and precipitation as inorganic P. In activated sludge treatment plants (AS), microbial P removal has been improved and is termed as enhanced biological phosphorus removal. There was a need to establish whether it was possible to enhance P removal in WSPs. A performance assessment of pond system at the University of Dare s Salaam (UDSM), Tanzania, has shown that 90% of the P removed was in the primary pond (facultative) and the rest in the maturation pond (aerobic). In these studies, a pure strain A. hydrophyla was isolated from an activated sludge wastewater treatment plant in Sweden. This plant has a train that functions with enhanced biological phosphorus removal. The strain was tested for P uptake in minimal media supplemented with glucose, succinate or acetate, grown aerobically and anaerobically/aerobically. This strain was able to take up P without having been subjected to the anaerobic phase. It was observed that P uptake was enhanced after the anaerobic phase with media supplemented with glucose, but not with succinate or acetate. Phosphorus uptake repeatedly followed the bacterial growth pattern with correlation coefficients of more than 95%. Therefore P removal has a direct correlation with bacterial growth. Two isolates Acinetobacter sp. (isolated from the primary facultative pond) and E .coli (isolated from the maturation pond) were obtained from a tropical WSP treatment system at the UDSM. They were subjected to aerobic P uptake experiment similar to those of A.hydrophyla. The uptake per unit absorbance of bacterial growth was found to be comparable to that of A.hydrophyla, isolated from AS. These results showed that, QC 20101119

Waste Stabilisation Ponds (WSPs) are characterised by low phosphorus (P) removal capacity. Heterotrophic bacteria are principal microbial agents in WSPs in addition to algae. As treatment proceeds in WSPs, algal growth increases and pH rises, this has lead to believe that P removal is mainly through sedimentation as organic P algal biomass and precipitation as inorganic P. In activated sludge treatment plants (AS), microbial P removal has been improved and is termed as enhanced biological phosphorus removal. There was a need to establish whether it was possible to enhance P removal in WSPs. A performance assessment of pond system at the University of Dare s Salaam (UDSM), Tanzania, has shown that 90% of the P removed was in the primary pond (facultative) and the rest in the maturation pond (aerobic). In these studies, a pure strain A. hydrophyla was isolated from an activated sludge wastewater treatment plant in Sweden. This plant has a train that functions with enhanced biological phosphorus removal. The strain was tested for P uptake in minimal media supplemented with glucose, succinate or acetate, grown aerobically and anaerobically/aerobically. This strain was able to take up P without having been subjected to the anaerobic phase. It was observed that P uptake was enhanced after the anaerobic phase with media supplemented with glucose, but not with succinate or acetate. Phosphorus uptake repeatedly followed the bacterial growth pattern with correlation coefficients of more than 95%. Therefore P removal has a direct correlation with bacterial growth. Two isolates Acinetobacter sp. (isolated from the primary facultative pond) and E .coli (isolated from the maturation pond) were obtained from a tropical WSP treatment system at the UDSM. They were subjected to aerobic P uptake experiment similar to those of A.hydrophyla. The uptake per unit absorbance of bacterial growth was found to be comparable to that of A.hydrophyla, isolated from AS. These results showed that, QC 20101119

Waste Stabilisation Ponds (WSPs) are characterised by low phosphorus (P) removal capacity. Heterotrophic bacteria are principal microbial agents in WSPs in addition to algae. As treatment proceeds in WSPs, algal growth increases and pH rises, this has lead to believe that P removal is mainly through sedimentation as organic P algal biomass and precipitation as inorganic P. In activated sludge treatment plants (AS), microbial P removal has been improved and is termed as enhanced biological phosphorus removal. There was a need to establish whether it was possible to enhance P removal in WSPs. A performance assessment of pond system at the University of Dare s Salaam (UDSM), Tanzania, has shown that 90% of the P removed was in the primary pond (facultative) and the rest in the maturation pond (aerobic). In these studies, a pure strain A. hydrophyla was isolated from an activated sludge wastewater treatment plant in Sweden. This plant has a train that functions with enhanced biological phosphorus removal. The strain was tested for P uptake in minimal media supplemented with glucose, succinate or acetate, grown aerobically and anaerobically/aerobically. This strain was able to take up P without having been subjected to the anaerobic phase. It was observed that P uptake was enhanced after the anaerobic phase with media supplemented with glucose, but not with succinate or acetate. Phosphorus uptake repeatedly followed the bacterial growth pattern with correlation coefficients of more than 95%. Therefore P removal has a direct correlation with bacterial growth. Two isolates Acinetobacter sp. (isolated from the primary facultative pond) and E .coli (isolated from the maturation pond) were obtained from a tropical WSP treatment system at the UDSM. They were subjected to aerobic P uptake experiment similar to those of A.hydrophyla. The uptake per unit absorbance of bacterial growth was found to be comparable to that of A.hydrophyla, isolated from AS. These results showed that, QC 20101119

Waste Stabilisation Ponds (WSPs) are characterised by low phosphorus (P) removal capacity. Heterotrophic bacteria are principal microbial agents in WSPs in addition to algae. As treatment proceeds in WSPs, algal growth increases and pH rises, this has lead to believe that P removal is mainly through sedimentation as organic P algal biomass and precipitation as inorganic P. In activated sludge treatment plants (AS), microbial P removal has been improved and is termed as enhanced biological phosphorus removal. There was a need to establish whether it was possible to enhance P removal in WSPs. A performance assessment of pond system at the University of Dare s Salaam (UDSM), Tanzania, has shown that 90% of the P removed was in the primary pond (facultative) and the rest in the maturation pond (aerobic). In these studies, a pure strain A. hydrophyla was isolated from an activated sludge wastewater treatment plant in Sweden. This plant has a train that functions with enhanced biological phosphorus removal. The strain was tested for P uptake in minimal media supplemented with glucose, succinate or acetate, grown aerobically and anaerobically/aerobically. This strain was able to take up P without having been subjected to the anaerobic phase. It was observed that P uptake was enhanced after the anaerobic phase with media supplemented with glucose, but not with succinate or acetate. Phosphorus uptake repeatedly followed the bacterial growth pattern with correlation coefficients of more than 95%. Therefore P removal has a direct correlation with bacterial growth. Two isolates Acinetobacter sp. (isolated from the primary facultative pond) and E .coli (isolated from the maturation pond) were obtained from a tropical WSP treatment system at the UDSM. They were subjected to aerobic P uptake experiment similar to those of A.hydrophyla. The uptake per unit absorbance of bacterial growth was found to be comparable to that of A.hydrophyla, isolated from AS. These results showed that, QC 20101119

This thesis describes the full-scale validation and calibration of a integrated metabolic activated sludge model for biological phosphorus removal. In chapters 1 and 2 the metabolic model is described, in chapters 3 to 6 the model is tested and in chapters 7 and 8 the model is put into practice. Chapter 1 is a general introduction to this research. Chapter 2 is a more specific introduction to the metabolic biological phosphorus removal (BioP) model. The goal of this introduction is to obtain a better understanding of the stoichiometric and kinetic structure of the metabolic model and the role of the storage polymers glycogen, poly-phosphate and PHB. In chapters 3 and 4, the model is validated at full-scale conditions. Chapter 3 describes the simulation of a full-scale nutrient removing WWTP at steady state conditions. In chapter 4, the start-up of a full-scale WWTP is simulated. Under start-up conditions model kinetics can be tested more extensively than is possible at steady state conditions. In chapter 5, a method presented for data evaluation, reconciliation and model calibration. The method is tested in a full-scale simulation study. In chapter 6, all previous modelling experiences with the metabolic BioP model are evaluated. The adapted model is tested on the basis of several lab-scale experiments. The updated version of the integrated model is presented in appendix III. In chapters 7 and 8, the model is put into practice. In a case study, a process control based on the oxidation reduction potential was evaluated. On the basis of a literature study, in chapter 7 the physical meaning of measuring the oxidation reduction potential in activated sludge is discussed. In chapter 8, it is demonstrated how the model can be used for (ORP related) process control and control design., Applied Sciences

This thesis describes the full-scale validation and calibration of a integrated metabolic activated sludge model for biological phosphorus removal. In chapters 1 and 2 the metabolic model is described, in chapters 3 to 6 the model is tested and in chapters 7 and 8 the model is put into practice. Chapter 1 is a general introduction to this research. Chapter 2 is a more specific introduction to the metabolic biological phosphorus removal (BioP) model. The goal of this introduction is to obtain a better understanding of the stoichiometric and kinetic structure of the metabolic model and the role of the storage polymers glycogen, poly-phosphate and PHB. In chapters 3 and 4, the model is validated at full-scale conditions. Chapter 3 describes the simulation of a full-scale nutrient removing WWTP at steady state conditions. In chapter 4, the start-up of a full-scale WWTP is simulated. Under start-up conditions model kinetics can be tested more extensively than is possible at steady state conditions. In chapter 5, a method presented for data evaluation, reconciliation and model calibration. The method is tested in a full-scale simulation study. In chapter 6, all previous modelling experiences with the metabolic BioP model are evaluated. The adapted model is tested on the basis of several lab-scale experiments. The updated version of the integrated model is presented in appendix III. In chapters 7 and 8, the model is put into practice. In a case study, a process control based on the oxidation reduction potential was evaluated. On the basis of a literature study, in chapter 7 the physical meaning of measuring the oxidation reduction potential in activated sludge is discussed. In chapter 8, it is demonstrated how the model can be used for (ORP related) process control and control design., Applied Sciences

The behaviour of a pilot scale biological phosphorus removal process (BPR) of the alternating type was investigated during periods of low influent concentrations and increased hydraulic load. A process disturbance of this type result in an increase in the phosphate concentration level in the anoxic/aerobic reactors and in the plant effluent shortlyafter the influent wastewater returns to normal strength. The accumulation of phosphorus in the system was avoided by the addition of an external carbon source either to the influent or to the effluent from the anaerobic reactor in form of sodium acetate. With the help of such an addition, the internal carbon storage compounds could be maintained at a high level, which is shown by poly-hydroxy-alcanoates (PHA) measurements. Several levels of acetate addition were investigated experimentallyin order to determine a minimal amount of internallystored carbon, which could ensure the stabilization of BPR during such dynamic influent conditions. Furthermore reduction of aeration time during periods of low influent concentrations was investigated. It was observed that BPR was stabilized bycombining a reduction of aeration time with carbon source addition, which maintained the internal stored carbon at a higher level. This combined control action resulted in a desired high BPR activitywhen the normal strength of the influent wastewater was reestablished. The failure of the BPR process was sometimes observed even when comparativelyhigh concentrations of PHA could be detected and an identification of a minimal PHA level was not possible. During this investigation an extended version of the activated sludge model No. 2 (ASM2), which includes denitrification byphosphate accumulating organisms, is used for the detailed analysis of the experiments. The model predicted the phosphorus build-up after the process disturbance as well as the performance during the stabilized experiments. Assisted by the model, the investigations indicate that

It is generally accepted that a decrease in temperature causes the rates of chemical and biochemical reactions to slow down, usually resulting in poorer performance of biological wastewater treatment systems. Despite this, early researchers repeatedly showed that excess biological phosphorus removal (EBPR) was more efficient at colder temperatures. Recent studies, however, have demonstrated that the reaction rates of EBPR processes decrease with temperature in accordance with Arrhenius' Law, resulting in an apparent contradiction in the literature. The objective of this study was to investigate the EBPR temperature controversy. The experimental systems used were two, lab-scale UCT configuration plants fed with acetate as the sole volatile fatty acid (VFA) source. The results showed that EBPR systems do perform more efficiently at colder temperatures, i.e., at 5°C compared to 20°C. The reason for better system performance was determined to be related to reduced competition for substrate in the non-oxic zones that results in an increased population of phosphate accumulating organisms (PAOs) relative to non-PAOs and, therefore, greater EBPR efficiency even though the reaction rates are slower. The proliferation of PAOs relative to non-PAOs at cold temperature indicates that some of the PAOs are psychrophilic, i.e., they have alternate biochemical pathways that give them a competitive advantage over bacteria dependent upon glycogen metabolism. The activated sludge acclimated to 20°C had relatively high polyhydroxyvalerate (PHV) and glycogen contents relative to sludge acclimated to 5°C. It was initially hypothized that there is a significant competition between PAO and glycogen accumulating organisms (GAOs) at 20°C and cold temperature (5°C) nearly eliminates this competition in favor of the PAOs. A series of batch test experiments revealed that despite similar acetate utilization by the sludges grown at the two temperatures nearly 30% less PHA was produced by the sludg

Although enhanced biological phosphorus removal (EBPR) and complete biological nutrient removal (BNR) systems can be operated successfully by experienced operators, the accuracy of design and strength of the scientific background need to be reinforced to enable accurate modeling and economically optimal design. One way to accomplish this would be through a better understanding of the biochemical mechanisms and microbial population dynamics that determine the reliability and efficiency of EBPR, and the utilization of this information to improve the design and operation of BNR plants. Such knowledge will also contribute to better structure of modeling tools that are used for design and educational purposes. The current body of knowledge is limited to observational studies that lack detailed biochemical explanations backed with a series of well planned experiments, and this has introduced uncertainties and inaccuracies into the biochemical and design models. Therefore, this study mainly covers a biochemical survey of the underlying metabolisms of active populations in BNR sludges. BNR biomass with biological phosphorus removal (BPR) capability was cultivated in continuous flow reactor (CFR) systems, configured as either University of Cape Town (UCT) and anoxic/oxic (A/O) systems. Following an acclimation period at 20°C, low temperature stress (5°C) was imposed on one UCT system for investigation of the response of the microbial consortium responsible from EBPR activity under cold temperature. Once a stable population with EBPR capabilities is established in each system, activities of ten enzymes that are hypothesized to be taking part in the EBPR metabolism were measured. These enzymes were selected among those that take part in major known pathways of bacterial energy and growth metabolism. Also, 13C-NMR was used as a tool to monitor the flux of labeled carbon in and out of pools of cellular storage; i.e. glycogen and polyhydroxyalkanoates (PHA). Combining the gathere

The laboratory-scale University of Cape Town (UCT) process was designed to investigate the effects of changing COD/TP ratios on the performance of biological nutrient removal (BNR) processes. Specific objectives of the research were to investigate the effects of COD/TP ratio on the rates of phosphorus removal, COD removal, nitrogen removal, PHB utilization and oxygen uptake. The system was fed with municipal wastewater and operated at 20° C. The influent COD concentration was held approximately constant while the phosphorus concentration was varied to obtained the desired COD/TP ratio. Once robust enhanced biological phosphorus removal (EBPR) has been established, the COD/TP ratios of 20, 30, 40 and 60 were investigated. The COD/TP ratio of the influent wastewater was observed to have a substantial effect upon the performance of the UCT BNR system. The amount of phosphorus removed by the system and the percent phosphorus in the aerobic zone MLVSS decreased as the COD/TP ratio increased. In addition, the amount of phosphorus released in the anaerobic zone per unit of COD removed in the anaerobic zone increased as the COD/TP ratio decreased. From this research, the amount of anaerobic COD removal required to remove 1 mg/L of phosphorus in the aerobic zone approached a minimum value as the COD/TP ratio decreased. It was also shown that PHB production increased as the COD/TP ratio increased. The highest specific oxygen uptake rate was always observed in the second aerobic reactor and tended to increase as the COD/TP ratio increased. However, the changes in the COD/TP ratio did not significant affect COD removal, nitrogen removal and the observed yield coefficient, but did strongly affect the MLSS concentration. The MLSS concentration at the COD/TP ratio of 60 was only 55% of that at the COD/TP ratio of 20. A high level of anaerobic COD removal, an elevated percent phosphorus in the waste activated sludge (WAS) and a high soluble effluent phosphorus concentration can be

The goal of this research was to characterize the effects of temperature changes on the denitrification process in a biological nutrient removal (BNR) system. Specifically, there were three objectives. First, the effects of temperature changes on denitrification rates by a bacterial population from a BNR system were investigated. Next, the role which PHAs (poly-beta-hydroxyalkanoates) played in the denitrification process were examined. Finally, the effect of temperature changes on the production and consumption rates of PHAs was determined. Sacrificial batch experiments were performed to assess the kinetic and chemical trends present in the denitrification process. Mixed liquor from the last anaerobic zone of a pilot scale BNR system was injected into vials. These vials were pre-purged with nitrogen gas in order to prevent dimolecular oxygen (02) from being entrained in the mixed liquor. Next, the vials were placed on a shaker table for 30 minutes in order to allow all external COD to be consumed. Following this, each vial was injected with nitrates and various macronutrients. This process was repeated for three different sets of batch tests; each set was identical except for the added substrate. One set received no added substrate while the other two received either acetate or glucose. Vials were sacrificed over a period of three hours and analyzed for nitrate, phosphate, PHB (polybeta-hydroxybutyrate), PHV (poly-beta-hydroxyvalerate), glucose and acetate.

Oxidation-Reduction Potential (ORP) can be used as a process control parameter for real-time automated control of batch sewage treatment systems. Results suggest that ORP has great potential for the control of anoxic sequences, since the distinct breakpoint in the ORP-time curve correlates to the disappearance of nitrates.

Oxidation-Reduction Potential (ORP) can be used as a process control parameter for real-time automated control of batch sewage treatment systems. Results suggest that ORP has great potential for the control of anoxic sequences, since the distinct breakpoint in the ORP-time curve correlates to the disappearance of nitrates.

Joint Research on Environmental Science and Technology for the Earth

Joint Research on Environmental Science and Technology for the Earth