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Ca. Accumulibacter was the predominant microorganism (relative FISH bio-abundance of 67 ± 5%) in a lab-scale sequential batch reactor that accomplished enhanced biological phosphorus removal (EBPR) while using glucose and acetate as the carbon sources (1:1 COD-based ratio). Both organic compounds were completely anaerobically consumed. The reactor's performance in terms of P/C ratio, phosphorous release and uptake, and overall kinetic and stoichiometric parameters were on the high end of the reported spectrum for EBPR systems (100:9.3 net mg phosphate removal per mg COD consumed when using glucose and acetate in a 1:1 ratio). The batch tests showed that, to the best of our knowledge, this is the first time a reactor enriched with Ca. Accumulibacter can putatively utilize glucose as the sole carbon source to biologically remove phosphate (COD:P (mg/mg) removal ratio of 100:6.3 when using only glucose). Thus, this research proposes that Ca. Accumulibacter directly anaerobically stored the fed glucose primarily as glycogen by utilizing the ATP provided via the hydrolysis of poly-P and secondarily as PHA by balancing its ATP utilization (glycogen generation) and formation (PHA storage). Alternative hypotheses are also discussed. The reported findings could challenge the conventional theories of glucose assimilation by Ca. Accumulibacter, and can be of significance for the biological removal of phosphorus from wastewaters with high contents of fermentable compounds or low VFAs.

Thermal drying is an effective sludge treatment method for dealing with large volumes of sludge. Microwave (MW) technology has been proposed as an effective and efficient technology for sludge drying. The physical-chemical properties of the sludge depend both on the origin of the sludge, as well as on the treatment process at which the sludge has been exposed. The physical-chemical properties of the sludge affect the performance and the subsequent valorisation and management of the sludge. This study evaluated the effect of certain physical-chemical properties of the sludge (moisture content, organic content, calorific value, porosity, hydrophobicity, and water-sludge molecular interaction, among others) on the MW sludge drying and energy performance. Four different types of sludge were evaluated collected from municipal wastewater treatment plants and septic tanks. The performance of the MW system was assessed by evaluating the sludge drying rates, exposure times, energy efficiencies and power input consumed by the MW system and linking the MW drying performance to the sludge physical-chemical properties. The results confirmed that MW drying substantially extends the constant drying period associated with unbound water evaporation, irrespective of the sludge sample evaluated. However, the duration and intensity were determined to depend on the dielectric properties of the sludge, particularly on the distribution of bound and free water. Sludge samples with a higher amount of free and loosely bound water absorbed and converted MW energy into heat more efficiently than sludge samples with a lower amount of free water. As a result, the sludge drying rates increased and the constant drying rate period prolonged; hence, leading to an increase in MW drying energy efficiency. The availability of free and loosely bound water molecules was favoured when hydrophobic compounds, e.g., oils and fats, were present in the sludge.

The operation of membrane bioreactors (MBRs) at higher than usual mixed liquor suspended solids (MLSS) concentrations may enhance the loading rate treatment capacity while minimizing even further the system's footprint. This requires operating the MBR at the highest possible MLSS concentration and biomass activity (e.g., at high loading rates and low solid retention times (SRTs)). Both a negative effect of the MLSS concentrations and a positive effect of the SRT on the oxygen transfer have been reported when using conventional fine bubble diffusers. However, most of the evaluations have been carried out either at extremely high SRTs or at low MLSS concentrations eventually underestimating the effects of the MLSS concentration on the oxygen transfer. This research evaluated the current limitations imposed by fine bubble diffusers in the context of the high-loaded MBR (HL-MBR) (i.e., high MLSS and short SRT-the latter emulated by concentrating municipal sludge from a wastewater treatment plant (WWTP) operated at a short SRT of approximately 5 days). The high MLSS concentrations and the short SRT of the original municipal sludge induced a large fraction of mixed liquor volatile suspended solids (MLVSS) in the sludge, promoting a large amount of sludge flocs that eventually accumulated on the surface of the bubbles and reduced the free water content of the suspension. Moreover, the short SRTs at which the original municipal sludge was obtained eventually appear to have promoted the accumulation of surfactants in the sludge mixture. This combination exhibited a detrimental effect on the oxygen transfer. Fine bubble diffusers limit the maximum MLSS concentration for a HL-MBR at 30 g L-1 ; beyond that point is either not technically or not economically feasible to operate ; an optimum MLSS concentration of 20 g L-1 is suggested to maximize the treatment capacity while minimizing the system's footprint.

The aerobic granular sludge (AGS) process is an effective wastewater treatment technology for organic matter and nutrient removal that has been introduced in the market rapidly. Until now, limited information is available on AGS regarding the removal of bacterial and viral pathogenic organisms present in sewage. This study focussed on determining the relation between reactor operational conditions (plug flow feeding, turbulent aeration and settling) and physical and biological mechanisms on removing two faecal surrogates, Escherichia coli and MS2 bacteriophages. Two AGS laboratory-scale systems were separately fed with influent spiked with 1.0 × 106 CFU/100 mL of E. coli and 1.3 × 108 PFU/100 mL of MS2 bacteriophages and followed during the different operational phases. The reactors contained only granular sludge and no flocculent sludge. Both systems showed reductions in the liquid phase of 0.3 Log10 during anaerobic feeding caused by a dilution factor and attachment of the organisms on the granules. Higher removal efficiencies were achieved during aeration, approximately 1 Log10 for E. coli and 0.6 Log10 for the MS2 bacteriophages caused mainly by predation. The 18S sequencing analysis revealed high operational taxonomic units (OTUs) of free-living protozoa genera Rhogostoma and Telotrochidium concerning the whole eukaryotic community. Attached ciliates propagated after the addition of the E. coli, an active contribution of the genera Epistylis, Vorticella, and Pseudovorticella was found when the reactor reached stability. In contrast, no significant growth of predators occurred when spiking the system with MS2 bacteriophages, indicating a low contribution of protozoa on the phage removal. Settling did not contribute to the removal of the studied bacterial and viral surrogates. M.L. Barrios-Hernández acknowledges the Technological Institute of Costa Rica for providing the fellowship (Grant Number 007-2014-M) to pursue her PhD programme (2016-2020) at IHE-Delft, the Netherlands. K. Mora-Cabrera acknowledges the Generalitat Valenciana (GRISOLIAP/2017/173) and the European Social Funds (BEFPI/2019/065) for their financial support.

The provision of effective sanitation strategies has a significant impact on public health. However, the treatment of septic sludge still presents some challenges worldwide. Consequently, innovative technologies capable of an effective and efficient sludge treatment, mostly at a decentralized level, are in high demand to improve sanitation provision. To address this problem, this study evaluates a novel semi-decentralised mobile faecal sludge treatment system, the pilot-system for which consists of a combination of several individual processes including mechanical dewatering (MD), microwave (MW) drying, and membrane filtration (ultrafiltration [UF] and reverse osmosis [RO]). The system evaluation was carried out by treating raw, partially digested faecal sludge (FS) from septic tanks—hence, septic sludge (SS)—in the Jordan Valley, Jordan. The pilot-scale system exhibited an effective and flexible treatment performance for (i) sanitizing faecal sludge and related liquid streams (MW and UF); (ii) reducing the treated sludge mass (and sludge volume) (MD and MW); and (iii) producing a high-quality treated liquid stream ideal for water reclamation applications (UF and RO). The MD process removed approximately 99% of the initial SS water content. The MW drying system completely removed E. coli and dehydrated the dewatered sludge at low energy expenditures of 0.75 MJ kg−1 and 5.5 MJ kg−1, respectively. Such energy expenditures can be further reduced by approximately 40% by recovering energy in the condensate and burning the dried sludge, which can then be reused inland applications. The membrane filtration system (UF and RO) was able to produce high-quality treated water that is ideal for the water reuse applications that irrigation requires, as well as meeting the Jordanian standard 893/2006. In addition, the system can also be powered by renewable energy sources, such as photovoltaic energy. Therefore, this research demonstrates that the evaluated semi-decentralised mobile system is technically feasible for the in situ treatment of SS (sanitization and dehydration), while also being effective for simultaneously recovering valuable resources, such as energy, water, and nutrients.

Photo-activated sludge (PAS) systems are an emerging wastewater treatment technology where microalgae provide oxygen to bacteria without the need for external aeration. There is limited knowledge on the optimal conditions for enhanced biological phosphorus removal (EBPR) in systems containing a mixture of polyphosphate accumulating organisms (PAOs) and microalgae. This research aimed to study the effects of substrate composition and light intensity on the performance of a laboratory-scale EBPR-PAS system. Initially, a model-based design was developed to study the effect of organic carbon (COD), inorganic carbon (HCO3) and ammonium-nitrogen (NH4-N) in nitrification deprived conditions on phosphorus (P) removal. Based on the mathematical model, two different synthetic wastewater compositions (COD:HCO3:NH4-N: 10:20:1 and 10:10:4) were examined at a light intensity of 350 µmol m−2 sec−1. Add to this, the performance of the system was also investigated at light intensities: 87.5, 175, and 262.5 µmol m−2 sec−1 for short terms. Results showed that wastewater having a high level of HCO3 and low level of NH4-N (ratio of 10:20:1) favored only microalgal growth, and had poor P removal due to a shortage of NH4-N for PAOs growth. However, lowering the HCO3 level and increasing the NH4-N level (ratio of 10:10:4) balanced PAOs and microalgae symbiosis, and had a positive influence on P removal. Under this mode of operation, the system was able to operate without external aeration and achieved a net P removal of 10.33 ±1.45 mg L−1 at an influent COD of 100 mg L−1. No significant variation was observed in the reactor performance for different light intensities, indicating the EBPR-PAS system can be operated at low light intensities with a positive influence on P removal.

The implementation of near-infrared (NIR) spectroscopy in the wastewater treatment has been continuously expanding. As an alternative to the conventional analytical methods for monitoring constituents in wastewater treatment processes, the use of NIR spectroscopy is considered cost-effective and less time- consuming. NIR spectroscopy does not in any way adulterate measured sample as no prior treatment is needed thus making it a waste-free technique. On the negative side, one must be very well acquainted with chemometric techniques to interpret results. In this study, filtered and centrifuged wastewater and sludge samples obtained from a lab-scale membrane bioreactor (MBR) were analysed. Essential wastewater constituents were determined and compared, employing two analytical methods (conventional and NIR spectroscopy). Special attention was given to the soluble microbial products (SMP) and the extracellular polymeric substances (EPS) which are known to promote membrane fouling. The measured parameters through NIR spectroscopy were analysed and processed by partial least squares regression (PLSR) and artificial neural networks (ANN) models to assess whether the evaluated wastewater constituents could be monitored by NIR spectroscopy. Very good results were obtained with PLSR models except for the determination of SMP thus limiting the model for their monitoring to qualitative rather than quantitative. ANN exhibited a better performance in terms of correlating NIR spectra with all the measured parameters, resulting in correlation coefficients in most cases higher than 0.97 for training, test, and validation. Based on the results achieved by this research, the combination of NIR spectra and chemometric modelling offers advantages compared to conventional analytical methods.

Conventional diffused aeration systems (such as fine-bubble diffusers) exhibit a poor oxygen transfer in wastewater treatment plants (WWTPs), particularly when operating at sludge concentrations higher than 15 g L−1. The supersaturated dissolved oxygen (SDOX) system has been proposed as an alternative for supplying dissolved oxygen (DO) at high mixed liquor suspended solids (MLSS) concentrations. The advantages introduced by such technology include the possibility of operating WWTPs at much higher than usual MLSS concentrations, increasing the treatment capacity of WWTPs. Recent studies have demonstrated that the SDOX system has higher oxygen transfer rates (OTRs) and oxygen transfer efficiencies (OTEs) relative to fine-bubble diffusers. However, it is unknown if the high-pressure conditions introduced by SDOX may possibly impact the biological performance of WWTPs. In this study, the effects of SDOX technology on the biological performance of a membrane bioreactor (MBR) were evaluated. The MBR was operated at an MLSS concentration of approximately 15 g L−1 in four phases as follows: (P1) with bubble diffusers, (P2) with an SDOX unit, (P3) with the bubble diffusers, and (P4) with the SDOX unit. The performance of the MBR was assessed by monitoring the sludge concentration, as well as changes in the particle size distribution (PSD), sludge activity, organic matter removal and nitrification performance, and changes in the microbial community within the MBR. The operational conditions exerted by the SDOX technology did not affect the concentration of active biomass during the study period. The biological performance of the MBR was not affected by the introduction of the SDOX technology. Finally, the microbial community was relatively stable although some variations at the family and genus level were evident during each of the study phases. Therefore, the SDOX system can be proposed as an alternative technology for DO supply in WWTPs increasing the overall treatment capacity.

The co-treatment of two synthetic faecal sludges (FS-1 and FS-2) with municipal synthetic wastewater (WW) was evaluated in an aerobic granular sludge (AGS) reactor. After characterisation, FS-1 showed the following concentrations, representative for medium-strength FS: 12,180 mg TSS L−1, 24,300 mg total COD L−1, 93.8 mg PO3-P L−1, and 325 mg NH4-N L−1. The NO3-N concentration was relatively high (300 mg L−1). For FS-2, the main difference with FS-1 was a lower nitrate concentration (18 mg L−1). The recipes were added consecutively, together with the WW, to an AGS reactor. In the case of FS-1, the system was fed with 7.2 kg total COD m−3d−1 and 0.5 kg Nitrogen m−3d−1. Undesired denitrification occurred during feeding and settling resulting in floating sludge and wash-out. In the case of FS-2, the system was fed with 8.0 kg total COD m−3d−1 and 0.3 kg Nitrogen m−3d−1. The lower NO3-N concentration in FS-2 resulted in less floating sludge, a more stabilised granular bed and better effluent concentrations. To enhance the hydrolysis of the slowly biodegradable particulates from the synthetic FS, an anaerobic stand-by period was added and the aeration period was increased. Overall, when compared to a control AGS reactor, a lower COD consumption (from 87 to 35 mg g−1 VSS h−1), P-uptake rates (from 6.0 to 2.0 mg P g VSS−1 h−1) and NH4-N removal (from 2.5 to 1.4 mg NH4-N g VSS−1 h−1) were registered after introducing the synthetic FS. Approximately 40% of the granular bed became flocculent at the end of the study, and a reduction of the granular size accompanied by higher solids accumulation in the reactor was observed. A considerable protozoa Vorticella spp. bloom attached to the granules and the accumulated particles occurred; potentially contributing to the removal of the suspended solids which were part of the FS recipe.

Sewage sludge management and treatment can represent up to approximately 30% of the overall operational costs of a wastewater treatment plant. Microwave (MW) drying has been recognized as a feasible technology for sludge treatment. However, MW drying systems exhibit high energy expenditures due to: (i) unnecessary heating of the cavity and other components of the system, (ii) ineffective extraction of the condensate from the irradiation cavity, and (iii) an inefficient use of the microwave energy, among others issues. This study investigated the performance of a novel pilot-scale MW system for sludge drying, specifically designed addressing the shortcomings previously described. The performance of the system was assessed drying municipal centrifuged wasted activated sludge at MW output powers from 1 to 6 kW and evaluating the system's drying rates and exposure times, specific energy outputs, MW generation efficiencies, overall energy efficiencies, and specific energy consumption. The results indicated that MW drying significantly extends the duration of the constant rate drying period associated with the evaporation of the unbound sludge water, a phase associated with low energy input requirement for evaporating water. Moreover, the higher the MW output power, the higher the sludge power absorption density, and the MW generation efficiency. MW generation efficiencies of up to 70% were reported. The higher the power absorption density, the lower the chances for energy losses in the form of reflected power and/or energy dissipated into the MW system. Specific energy consumptions as low as 2.6 MJ L−1 (0.74 kWh L−1) could be achieved, well in the range of conventional thermal dryers. The results obtained in this research provide sufficient evidence to conclude that the modifications introduced to the novel pilot-scale MW system mitigated the shortcomings of existing MW systems, and that the technology has great potential to effectively and efficiently drying municipal sewage sludge.

Populations of “Candidatus Accumulibacter”, a known polyphosphate-accumulating organism, within clade IC have been proposed to perform anoxic P-uptake activity in enhanced biological phosphorus removal (EBPR) systems using nitrate as electron acceptor. However, no consensus has been reached on the ability of “Ca. Accumulibacter” members of clade IC to reduce nitrate to nitrite. Discrepancies might relate to the diverse operational conditions which could trigger the expression of the Nap and/or Nar enzyme and/or to the accuracy in clade classification. This study aimed to assess whether and how certain operational conditions could lead to the enrichment and enhance the denitrification capacity of “Ca. Accumulibacter” within clade IC. To study the potential induction of the denitrifying enzyme, an EBPR culture was enriched under anaerobic–anoxic–oxic (A2O) conditions that, based on fluorescence in situ hybridization and ppk gene sequencing, was composed of around 97% (on a biovolume basis) of affiliates of “Ca. Accumulibacter” clade IC. The influence of the medium composition, sludge retention time (SRT), polyphosphate content of the biomass (poly-P), nitrate dosing approach, and minimal aerobic SRT on potential nitrate reduction were studied. Despite the different studied conditions applied, only a negligible anoxic P-uptake rate was observed, equivalent to maximum 13% of the aerobic P-uptake rate. An increase in the anoxic SRT at the expenses of the aerobic SRT resulted in deterioration of P-removal with limited aerobic P-uptake and insufficient acetate uptake in the anaerobic phase. A near-complete genome (completeness = 100%, contamination = 0.187%) was extracted from the metagenome of the EBPR biomass for the here-proposed “Ca. Accumulibacter delftensis” clade IC. According to full-genome-based phylogenetic analysis, this lineage was distant from the canonical “Ca. Accumulibacter phosphatis”, with closest neighbor “Ca. Accumulibacter sp. UW-LDO-IC” within clade IC. This was cross-validated with taxonomic classification of the ppk1 gene sequences. The genome-centric metagenomic analysis highlighted the presence of genes for assimilatory nitrate reductase (nas) and periplasmic nitrate reductase (nap) but no gene for respiratory nitrate reductases (nar). This suggests that “Ca. Accumulibacter delftensis” clade IC was not capable to generate the required energy (ATP) from nitrate under strict anaerobic-anoxic conditions to support an anoxic EBPR metabolism. Definitely, this study stresses the incongruence in denitrification abilities of “Ca. Accumulibacter” clades and reflects the true intra-clade diversity, which requires a thorough investigation within this lineage.

The high frequency and intensity of urban floods caused by climate change, urbanisation and infrastructure failures increase public health risks when the flood water contaminated from combined sewer overflows (CSOs) or other sources of faecal contamination remains on urban surfaces. This study contributes to a better understanding of the effects of urban and recreational surfaces on the occurrence of waterborne pathogens. The inactivation of selected indicator organisms was studied under controlled exposure to artificial sunlight for 6 h followed by 18 h in dark conditions. Concrete, asphalt, pavement blocks and glass as control were inoculated with artificial floodwater containing, as indicator organisms, Escherichia coli bacteria, which are common faecal indicator bacteria (FIB) for water quality assessment, Bacillus subtilis spores chosen as surrogates for Cryptosporidium parvum oocysts and Giardia cysts, and bacteriophages MS2 as indicators for viral contamination. On practically all the surfaces in this study, E. coli had the highest inactivation under light conditions followed by MS2 and B. subtilis, except asphalt where MS2 was inactivated faster. The highest inactivation under light conditions was seen with E. coli on a concrete surface (pH 9.6) with an inactivation rate of 1.85 h−1. However, the pH of the surfaces (varying between 7.0 and 9.6) did not have any influence on inactivation rates under dark conditions. MS2 bacteriophage had the highest inactivation under light conditions on asphalt with a rate of 1.29 h−1. No die-off of B. subtilis spores was observed on any of the surfaces during the experiment, neither in light nor in dark conditions. This study underpins the need to use different indicator organisms to test their inactivation after flooding. It also suggests that given the sunlight conditions, concentration of indicator organisms and type of surface, the fate of waterborne pathogens after a flood could be estimated.

A laboratory-scale microwave (MW) unit was applied to treat fresh blackwater sludge that represented fecal sludge (FS) produced at heavily used toilet facilities. The sludge was exposed to MW irradiation at different power levels and for various durations. Variables such as sludge volume and pathogen reduction were observed. The results demonstrated that the MW is a rapid and efficient technology that can reduce the sludge volume by over 70% in these experimental conditions. The concentration of bacterial pathogenic indicator E. coli also decreased to below the analytical detection levels. Furthermore, the results indicated that the MW operational conditions including radiation power and contact time can be varied to achieve the desired sludge volume and pathogen reduction. MW technology can be further explored for the potential scaling-up as an option for rapid treatment of FS from intensively used sanitation facilities such as in emergency situations., Graphical abstract, Highlights • There is lack of fast and efficient fecal sludge treatment options in emergencies. • Microwave treatment is rapid and efficient in sludge volume and pathogen reduction. • Power and contact time can be varied to reach diverse levels of sludge treatment.

The aim of this study was to evaluate the effectiveness of the novel aerobic granular sludge (AGS) wastewater treatment technology in removing faecal indicator organisms (FIOs) compared to the conventional activated sludge (CAS) treatment system. The work was carried out at two full-scale wastewater treatment plants (WWTP) in the Netherlands, Vroomshoop and Garmerwolde. Both treatment plants have a CAS and AGS system operated in parallel. The parallel treatment lines are provided with the same influent wastewater. The concentrations of the measured FIOs in the influent of the two WWTPs were comparable with reported literature values as follows: F-specific RNA bacteriophages at 106 PFU/100 mL, and Escherichia coli (E. coli), Enterococci, and Thermotolerant coliforms (TtC) at 105 to 106 CFU/100 mL. Although both systems (CAS and AGS) are different in terms of design, operation, and microbial community, both systems showed similar FIOs removal efficiency. At the Vroomshoop WWTP, Log10 removals for F-specific RNA bacteriophages of 1.4 ± 0.5 and 1.3 ± 0.6 were obtained for the AGS and CAS systems, while at the Garmerwolde WWTP, Log10 removals for F-specific RNA bacteriophages of 1.9 ± 0.7 and 2.1 ± 0.7 were found for the AGS and CAS systems. Correspondingly, E. coli, Enterococci, and TtC Log10 removals of 1.7 ± 0.7 and 1.1 ± 0.7 were achieved for the AGS and CAS systems at Vroomshoop WWTP. For Garmerwolde WWTP Log10 removals of 2.3 ± 0.8 and 1.9 ± 0.7 for the AGS and CAS systems were found, respectively. The measured difference in removal rates between the plants was not significant. Physicochemical water quality parameters, such as the concentrations of organic matter, nutrients, and total suspended solids (TSS) were also determined. Overall, it was not possible to establish a direct correlation between the physicochemical parameters and the removal of FIOs for any of the treatment systems (CAS and AGS). Only the removal of TSS could be positively correlated to the E. coli removal for the AGS technology at the evaluated WWTPs., Graphical abstract Image 1, Highlights • No difference in removal efficiencies (α = 5%) of FIOs was found for the studied AGS and CAS systems. • 1.3 to 2.1 Log10 removals for F-specific RNA bacteriophages were found for the CAS and AGS systems. • 1.1 to 2.3 Log10 removals for E. coli, Enterococci, and TtC were obtained for the AGS and CAS systems. • The two stage CAS performed better than the carrousel CAS in removing F-specific RNA bacteriophages. • A positive correlation was found between the TSS and E. coli removal, and COD and TtC removal in the AGS systems.

The occurrence of Accumulibacter Type I (a known phosphorus-accumulating organism, PAO) has received increased attention due to the potential operating benefits associated with their denitrifying activity in enhanced biological phosphorus removal (EBPR) wastewater treatment plants. In this study, after a shift from an enriched glycogen-accumulating organism (GAO) culture (competitors of PAO) to a PAO-enriched system, Accumulibacter Type I (PAO I) became dominant in an anaerobic-aerobic EBPR system fed with acetate and operated at 10°C with a net aerobic solids retention time (SRT) of 6 d. Since Accumulibacter Type II (PAO II) were not detected, the low temperature in combination with the net aerobic SRT applied appeared to have suppressed their growth as well. The stoichiometry of PAO I was in agreement with previous metabolic models, suggesting that it was the main PAO organisms present in previous studies operated under similar conditions. Moreover, under poly-P limiting conditions, PAO I were unable to switch to a GAO-like metabolism at low temperatures. These results contribute to increase the understanding of the physiology, microbial metabolism and microbial ecology of PAO I.

The overall performance of the Anjana wastewater treatment plant (WWTP) located in Surat, India, was assessed by coupling the Activated Sludge Model No. 3 (ASM3) and the Anaerobic Digestion Model No. 1 (ADM1). Guidelines developed by the Dutch Foundation for Applied Water Research (STOWA) were successfully applied for the determination of wastewater characteristics. Concerning the fractionation of primary and secondary sludge, the approach proved to be adequate for the application of ADM1. A satisfactory description of the performance of the plant was obtained in terms of effluent quality, biogas generation and sludge production. This was achieved through coupling ASM3 with ADM1 and adjusting four default values (the growth of autotrophic bacteria from 1 to 0.46 day−1, influent fraction of unbiodegradable particulate chemical oxygen demand (COD) to 0.14 gCOD/gCOD, and the anaerobic disintegration factors for soluble and particulate unbiodegradable COD in ADM1 to 0.01 and 0.29 gCOD/gCOD, respectively). The model was applied to optimise the plant performance and to assess the potential influence of the return of high strength reject effluents through the implementation of an ADM1-ASM3 interface. This study underlines the feasibility, advantages and benefits of mathematical modelling as a reliable tool for process optimisation, plant upgrade and resource recovery in developing countries.

Although simultaneous P-removal and nitrate reduction has been observed in laboratory studies as well as full-scale plants, there are contradictory reports on the ability of PAO I to efficiently use nitrate as electron acceptor. Such discrepancy could be due to other microbial groups performing partial denitrification from nitrate to nitrite. The denitrification capacities of two different cultures, a highly enriched PAO I and a PAO I-GAO cultures were assessed through batch activity tests conducted before and after acclimatization to nitrate. Negligible anoxic phosphate uptake coupled with a reduction of nitrate was observed in the highly enriched PAO I culture. On the opposite, the PAO I-GAO culture showed a higher anoxic phosphate uptake activity. Both cultures exhibited good anoxic phosphate uptake activity with nitrite (8.7 ± 0.3 and 9.6 ± 1.8 mgPO4-P/gVSS.h in the PAO I and PAO I-GAO cultures, respectively). These findings suggest that other microbial populations, such as GAOs, were responsible to reduce nitrate to nitrite in this EBPR system, and that PAO I used the nitrite generated for anoxic phosphate uptake. Moreover, the simultaneous denitrification and phosphate removal process using nitrite as electron acceptor may be a more sustainable process as can: i) reduce the carbon consumption, ii) reduce oxygen demand of WWTP, and iii) due to a lower growth yield contribute to a lower sludge production.

Phosphate accumulating organisms (PAO) are assumed to use nitrate as external electron acceptor, allowing an efficient integration of simultaneous nitrogen and phosphate removal with minimal organic carbon (COD) requirements. However, contradicting findings appear in literature regarding the denitrification capacities of PAO due to the lack of clade specific highly enriched PAO cultures. Whereas some studies suggest that only PAO clade I may be capable of using nitrate as external electron acceptor for anoxic P-uptake, other studies indicate that PAO clade II may be responsible for anoxic P-removal. In the present study, a highly enriched PAO clade IC culture (>99% according to FISH) was cultivated in an SBR operated under Anaerobic/Oxic conditions and subsequently exposed to Anaerobic/Anoxic/Oxic conditions using nitrate as electron acceptor. Before and after acclimatization to the presence of nitrate, the aerobic and anoxic (nitrate and nitrite) activities of the PAO I culture were assessed through the execution of batch tests using either acetate or propionate as electron donor. In the presence of nitrate, significant P-uptake by PAO I was not observed before or after acclimatization. Using nitrite as electron acceptor, limited nitrite removal rates were observed before acclimatization with lower rates in the acetate fed reactor without P-uptake and slightly higher in the propionate fed reactor with a marginal anoxic P-uptake. Only after acclimatization to nitrate, simultaneous P and nitrite removal was observed. This study suggests that PAO clade IC is not capable of using nitrate as external electron acceptor for anoxic P-removal. The elucidation of the metabolic capacities for individual PAO clades helps in better understanding and optimization of the relation between microbial ecology and process performance in enhanced biological phosphate removal processes.

The influence of different carbon sources (acetate to propionate ratios), temperature and pH levels on the competition between polyphosphate- and glycogen-accumulating organisms (PAO and GAO, respectively) was evaluated using a metabolic model that incorporated the carbon source, temperature and pH dependences of these microorganisms. The model satisfactorily described the bacterial activity of PAO (Accumulibacter) and GAO (Competibacter and Alphaproteobacteria-GAO) laboratory-enriched cultures cultivated on propionate (HPr) and acetate (HAc) at standard conditions (20 degrees C and pH 7.0). Using the calibrated model, the effects of different influent HAc to HPr ratios (100-0, 75-25, 50-50 and 0-100%), temperatures (10, 20 and 30 degrees C) and pH levels (6.0, 7.0 and 7.5) on the competition among Accumulibacter, Competibacter and Alphaproteobacteria-GAO were evaluated. The main aim was to assess which conditions were favorable for the existence of PAO and, therefore, beneficial for the biological phosphorus removal process in sewage treatment plants. At low temperature (10 degrees C), PAO were the dominant microorganisms regardless of the used influent carbon source or pH. At moderate temperature (20 degrees C), PAO dominated the competition when HAc and HPr were simultaneously supplied (75-25 and 50-50% HAc to HPr ratios). However, the use of either HAc or HPr as sole carbon source at 20 degrees C was not favorable for PAO unless a high pH was used (7.5). Meanwhile, at higher temperature (30 degrees C), GAO tended to be the dominant microorganisms. Nevertheless, the combined presence of acetate and propionate in the influent (75-25 and 50-50% HAc to HPr ratios) as well as a high pH (7.5) appear to be potential factors to favor the metabolism of PAO over GAO at higher sewage temperature (30 degrees C).

Ultraviolet germicidal (short wavelength UV-C) light was studied as surface disinfectant in an Emergency Sanitation Operation System(®) smart toilet to aid to the work of manual cleaning. The UV-C light was installed and regulated as a self-cleaning feature of the toilet, which automatically irradiate after each toilet use. Two experimental phases were conducted i.e. preparatory phase consists of tests under laboratory conditions and field testing phase. The laboratory UV test indicated that irradiation for 10 min with medium-low intensity of 0.15-0.4 W/m(2) could achieve 6.5 log removal of Escherichia coli. Field testing of the toilet under real usage found that UV-C irradiation was capable to inactivate total coliform at toilet surfaces within 167-cm distance from the UV-C lamp (UV-C dose between 1.88 and 2.74 mW). UV-C irradiation is most effective with the support of effective manual cleaning. Application of UV-C for surface disinfection in emergency toilets could potentially reduce public health risks.

The anaerobic acetate (HAc) uptake stoichiometry of phosphorus-accumulating organisms (PAO) in enhanced biological phosphorus removal (EBPR) systems has been an extensive subject of study due to the highly variable reported stoichiometric values (e.g. anaerobic P-release/HAc-uptake ratios ranging from 0.01 up to 0.93 P-mol/C-mol). Often, such differences have been explained by the different applied operating conditions (e.g. pH) or occurrence of glycogen-accumulating organisms (GAO). The present study investigated the ability of biomass highly enriched with specific PAO clades (‘Candidatus Accumulibacter phosphatis’ Clade I and II, hereafter PAO I and PAO II) to adopt a GAO metabolism. Based on long-term experiments, when Poly-P is not stoichiometrically limiting for the anaerobic VFA uptake, PAO I performed the typical PAO metabolism (with a P/HAc ratio of 0.64 P-mol/C-mol); whereas PAO II performed a mixed PAO-GAO metabolism (showing a P/HAc ratio of 0.22 P-mol/C-mol). In short-term batch tests, both PAO I and II gradually shifted their metabolism to a GAO metabolism when the Poly-P content decreased, but the HAc-uptake rate of PAO I was 4 times lower than that of PAO II, indicating that PAO II has a strong competitive advantage over PAO I when Poly-P is stoichiometrically limiting the VFA uptake. Thus, metabolic flexibility of PAO clades as well as their intrinsic differences are additional factors leading to the controversial anaerobic stoichiometry and kinetic rates observed in previous studies. From a practical perspective, the dominant type of PAO prevailing in full-scale EBPR systems may affect the P-release processes for biological or combined biological and chemical P-removal and recovery and consequently the process performance.

In this study the following was evaluated: (a) application of a complex activated sludge model on a full-scale plant (Phostrip ® -like process), (b) influent and sludge characterization procedures for bio-P modeling, (c) the use of batch tests for model evaluation and (d) different alternative BPR process configurations (A/O, UCT and BCFS ® ). An integrated model for aerobic and denitrifying biological phosphorus removal (Delft bio-P model) replaced the module for P removal of ASM No. 2 and was combined with retained equations for COD and N conversion of the ASM No 2. This combined model proved well capable of describing the wastewater treatment plant (wwtp) Haarlem Waarderpolder with adjustment of only three (out of the sixty) default parameters. Some of batch tests were satisfactorily described by the model too. Batch tests proved as a useful tool for sludge characterization and model validation. The standard Dutch procedure for influent and sludge characterization proved satisfactory for the model construction. Modeling of alternative BPR process configuration showed that good P removal was achieved by all three alternative process configurations, by the latest one with comparatively lowest construction and operational costs (lower building costs and no acetate addition). Valuable experience from practical application of the model was obtained; study indicated where the model should be improved; the plant operation and treatment processes were better understood and plant performance was further optimized.

This paper describes the impact of long-term (weeks) temperature changes on stoichiometry and kinetics of the anaerobic and aerobic phases of the biological phosphorus removal process. Steady state conversion of relevant compounds for biological phosphorus removal was studied at 20, 30, 20, 10 and 5°C, following chronological order. Integrated in the process study, two methods (electron-microscopy and dry denaturing gradient gel electrophoresis) were applied to investigate the complexity of the bacterial community of biological phosphorus removing sludge cultivated at different temperatures. The coefficient for metabolic conversions obtained from long-term temperature tests was similar to the temperature coefficient observed in short-term (hours) tests ( θ =1.085 versus θ =1.078, respectively).Temperature had a moderate impact on the aerobic P-uptake process rate ( θ =1.031) during long-term tests. However, a strong temperature effect on other metabolic processes of the aerobic phase, such as polyhydroxyalkanoate consumption ( θ =1.163), oxygen uptake ( θ =1.090) and growth ( θ >1.110), was observed. Different temperature coefficients were obtained for the aerobic phase from long-term and short-term tests, probably due to a change in population structure. This change was also visible from molecular ecological studies. The different temperature coefficient found for P-uptake compared to the other metabolic processes of the aerobic phase underlines that, in complex processes such as BPR, it is dangerous to draw conclusions from easily observable parameters (like phosphate) only. Such consideration can easily lead to underestimation of the temperature dependency of other metabolic processes of the aerobic phase of BPR.

It has been reported that deterioration of biological phosphorus removal (BPR) efficiency at some wastewater treatment plants (WWTPs) regularly occurred after heavy rainfall or weekends. The deterioration has been attributed to low plant loading that took place during such events. However, it is hypothesized in this study that the cause of such deterioration may have been the excessive aeration that took place at some of those plants due to inadequate control of aeration system during weekends and rainfall periods. In order to prove this hypothesis, the influence of excessive aeration (aeration during starvation conditions) on BPR processes was studied using a laboratory anaerobic-aerobic-settling sequencing batch reactor (SBR). It was clearly demonstrated that the phosphorus uptake stops due to a gradual depletion of poly-hydroxy-butyrate (PHB) in an over-aerated process. If organic substrate is introduced to the system, phosphorus release is immediately at its maximal rate. However, the released phosphorus cannot be taken-up fully again because the PHB content limits the uptake rate. Consequently, incomplete phosphorus uptake leads to temporary reduction of BPR efficiency. This causal effect can explain the deterioration of BPR efficiency after heavy rainfall or weekends. Since excessive aeration clearly negatively affects the BPR processes, the aeration should be properly controlled at sewage treatment plants. Some other findings of this study deserve to be mentioned. • • It was confirmed that the presence of acetate under aerobic conditions provokes phosphorus release. This may also contribute to deterioration of the BPR efficiency. • • The aerobic phosphate uptake was found to depend not only on the PHB but also on polyphosphate (poly-P) content of the cells. • • A maximal poly-P (0.18 g-P/g-VSS) and minimal PHB content of the cells (2.11 mg-COD/g-VSS) were observed in the enriched sludge during excessive aeration experiments. • • It was shown that, under aerobic starvation conditions, glycogen can not replace PHB for phosphate uptake and is only used for maintenance. During this period, no oxygen consumption due to decay processes has been observed. © 1998 Elsevier Science Ltd. All rights reserved

Hong Kong has practiced seawater toilet flushing since 1958, saving 750,000 m3 of freshwater every day. A high sulfate-to-COD ratio (>1.25 mg SO 4 2 − /mg COD) in the saline sewage resulting from this practice has enabled us to develop the S ulfate reduction, A utotrophic denitrification and N itrification I ntegrated (SANI®) process with minimal sludge production and oxygen demand. Recently, the SANI® process has been expanded to include Enhanced Biological Phosphorus Removal (EBPR) in an alternating anaerobic/limited-oxygen (LOS-EBPR) aerobic sequencing batch reactor (SBR). This paper presents further development – an anaerobic/anoxic denitrifying sulfur cycle-associated EBPR, named as DS-EBPR, bioprocess in an alternating anaerobic/anoxic SBR for simultaneous removal of organics, nitrogen and phosphorus. The 211 day SBR operation confirmed the sulfur cycle-associated biological phosphorus uptake utilizing nitrate as electron acceptor. This new bioprocess cannot only reduce operation time but also enhance volumetric loading of SBR compared with the LOS-EBPR. The DS-EBPR process performed well at high temperatures of 30 °C and a high salinity of 20% seawater. A synergistic relationship may exist between sulfur cycle and biological phosphorus removal as the optimal ratio of P-release to SO 4 2 − -reduction is close to 1.0 mg P/mg S. There were no conventional PAOs in the sludge.

A full-scale sewage treatment plant designed for biological phosphorus removal may experience short- or long-term shortage in potassium of the influent. In this study, using an anaerobic-aerobic sequenced batch reactor system, inoculation sludge from laboratory-, pilot- and full-scale phosphorus removal plants was exposed to different potassium-phosphorus ratios in the influent. By simulating the conditions which may occur in practice, it was shown that potassium is an essential factor in biological phosphorus removal processes. When the system was exposed to severe shortage of potassium in the influent: (a) phosphorus removal was absent, (b) polyphosphate concentration in the biomass decreased exponentially due to sludge wasting and (c) the anaerobic phosphorus release and the related acetate uptake was only affected after several days of potassium absence, likely due to insufficient content of polyphosphate in the biomass to allow full acetate uptake under anaerobic conditions. In contrast, the system achieved complete phosphorus removal when potassium was present in excess amounts.

Hong Kong has practiced seawater toilet flushing since 1958, saving 750,000 m 3 freshwater every day. A high sulfate-to-COD ratio (>1.25 mg SO 4 /mg COD) in the saline sewage resulting from this practice has enabled us to develop the S ulfate reduction A utotrophic denitrification and Nitrification I ntegrated (SANI ® ) process with minimal sludge production. This study seeks to expand the SANI process into an enhanced biological phosphorus removal (EBPR) process. A sulfur cycle associated EBPR was explored in an alternating anaerobic/oxygen-limited aerobic sequencing batch reactor with acetate fed as sole electron donor and sulfate as sulfur source at a total organic carbon to sulfur ratio of 1.1–3.1 (mg C/mg S). Phosphate uptake and polyphosphate formation was observed in this reactor that sustained high phosphate removal (20 mg P/L removed with 320 mg COD/L). This new EBPR process was supported by six observations: 1) anaerobic phosphate release associated with acetate uptake, poly-phosphate hydrolysis, poly-hydroxyalkanoate (PHA) (and poly-S 2− /S 0 ) formation and an “aerobic” phosphate uptake associated with PHA (and poly-S 2− /S 0 ) degradation, and polyphosphate formation; 2) a high P/VSS ratio (>0.16 mg P/mg VSS) and an associated low VSS/TSS ratio (0.75) characteristic of conventional PAOs; 3) a lack of P-release and P-uptake with formaldehyde inactivation and autoclaved sterilized biomass; 4) an absence of chemical precipitated P crystals as determined by XRD analysis; 5) a sludge P of more than 90% polyphosphate as determined by sequential P extraction; and 6) microscopically, observed PHA, poly-P and S globules in the biomass.

Enhanced biological phosphorus removal (EBPR) from wastewater relies on the enrichment of activated sludge with phosphorus-accumulating organisms (PAOs). The presence and proliferation of glycogen-accumulating organisms (GAOs), which compete for substrate with PAOs, may be detrimental for EBPR systems, leading to deterioration and, in extreme cases, failure of the process. Therefore, from both process evaluation and modeling perspectives, the estimation of PAO and GAO populations in activated sludge systems is a relevant issue. A simple method for the quantification of PAO and GAO population fractions in activated sludge systems is presented in this paper. To develop such a method, the activity observed in anaerobic batch tests executed with different PAO/GAO ratios, by mixing highly enriched PAO and GAO cultures, was studied. Strong correlations between PAO/GAO population ratios and biomass activity were observed (R2 > 0.97). This served as a basis for the proposal of a simple and practical method to quantify the PAO and GAO populations in activated sludge systems, based on commonly measured and reliable analytical parameters (i.e., mixed liquor suspended solids, acetate, and orthophosphate) without requiring molecular techniques. This method relies on the estimation of the total active biomass population under anaerobic conditions (PAO plus GAO populations), by measuring the maximum acetate uptake rate in the presence of excess acetate. Later, the PAO and GAO populations present in the activated sludge system can be estimated, by taking into account the PAO/GAO ratio calculated on the basis of the anaerobic phosphorus release-to-acetate consumed ratio. The proposed method was evaluated using activated sludge from municipal wastewater treatment plants. The results from the quantification performed following the proposed method were compared with direct population estimations carried out with fluorescence in situ hybridization analysis (determining Candidatus Accumulibacter Phosphatis as PAO and Candidatus Competibacter Phosphatis as GAO). The method showed to be potentially suitable to estimate the PAO and GAO populations regarding the total PAO-GAO biomass. It could be used, not only to evaluate the performance of EBPR systems, but also in the calibration of potential activated sludge mathematical models, regarding the PAO-GAO coexistence.

The influence of operating and environmental conditions on the microbial populations of the enhanced biological phosphorus removal (EBPR) process at seven full-scale municipal activated sludge wastewater treatment plants (WWTPs) in The Netherlands was studied. Data from the selected WWTPs concerning process configuration, operating and environmental conditions were compiled. The EBPR activity from each plant was determined by execution of anaerobic-anoxic-aerobic batch tests using fresh activated sludge. Fractions of Accumulibacter as potential phosphorus accumulating organisms (PAO), and Competibacter, Defluviicoccus-related microorganisms and Sphingomonas as potential glycogen accumulating organisms (GAO) were quantified using fluorescence in situ hybridization (FISH). The relationships among plant process configurations, operating parameters, environmental conditions, EBPR activity and microbial populations fractions were evaluated using a statistical approach. A well-defined and operated denitrification stage and a higher mixed liquor pH value in the anaerobic stage were positively correlated with the occurrence of Accumulibacter. A well-defined denitrification stage also stimulated the development of denitrifying PAO (DPAO). A positive correlation was observed between Competibacter fractions and organic matter concentrations in the influent. Nevertheless, Competibacter did not cause a major effect on the EBPR performance. The observed Competibacter fractions were not in the range that would have led to EBPR deterioration. Likely, the low average sewerage temperature (12+/-2 degrees C) limited their proliferation. Defluviicoccus-related microorganisms were seen only in negligible fractions in a few plants (

In biological nitrogen removal processes, nitrite can be formed and accumulated through both nitrification and denitrification. Despite the fact that, in practice, biological phosphate removal (BPR) is often combined with biological nitrogen removal, there are only a few publications reporting the effect of nitrite on BPR. In this study, phosphate-accumulating organisms (PAO) were cultivated in an anaerobic–anoxic–aerobic sequencing batch reactor (SBR). The effect of nitrite on the enrichment of the sludge with PAO, the phosphate uptake rates and the sludge respiration was investigated. The results indicate that (1) presence of nitrite inhibits both aerobic and anoxic (denitrifying) phosphate uptake, (2) aerobic phosphate uptake was more affected than anoxic phosphate uptake, (3) presence of nitrite could be one of the factors enhancing the presence of glycogen accumulating organisms (GAO)—competitors to PAO for substrate in the anaerobic phase, and (4) it is required to monitor and control nitrite accumulation in a full-scale wastewater treatment plants.