Influence of the Sludge Retention Time on Membrane Fouling in an Anaerobic Membrane Bioreactor (AnMBR) Treating Lipid-Rich Dairy Wastewater.

This study evaluated the effects of sludge retention time (SRT) on the membrane filtration performance of an anaerobic membrane bioreactor (AnMBR) fed lipid-rich synthetic dairy wastewater. The membrane filtration performance was evaluated in two AnMBR systems operated at two different SRTs, i.e., 20 and 40 days. For the AnMBR operated at 40 days, SRT exhibited worse membrane filtration performance characterized by operational transmembrane pressures (TMP) exceeding the maximum allowed value and high total resistances to filtration (Rtotal). The sludge in the two reactors evaluated at the different SRTs showed similar sludge filterability properties. However, the sludge in the reactor operated at 40 days SRT was characterized by exhibiting the highest concentrations of: (i) total suspended solids (TSS), (ii) small-sized particles, (iii) extracellular polymeric substances (EPS), (iv) soluble microbial products (SMP), (v) fats, oils and grease (FOG), and (vi) long-chain fatty acids (LCFA). The cake layer resistance was the major contributor to the overall resistance to filtration. The high TSS concentration observed in the AnMBR systems apparently contributed to a less permeable cake layer introducing a negative effect on the membrane filtration performance.

Effects of the sludge physical-chemical properties on its microwave drying performance.

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.

Novel semi-decentralised mobile system for the sanitization and dehydration of septic sludge: a pilot-scale evaluation in the Jordan Valley.

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.

Unravelling the removal mechanisms of bacterial and viral surrogates in aerobic granular sludge systems.

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.

Effect of the co-treatment of synthetic faecal sludge and wastewater in an aerobic granular sludge system.

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.

Microwave treatment of municipal sewage sludge: Evaluation of the drying performance and energy demand of a pilot-scale microwave drying system.

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.

Removal of bacterial and viral indicator organisms in full-scale aerobic granular sludge and conventional activated sludge systems.

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.

Sidestream superoxygenation for wastewater treatment: Oxygen transfer in clean water and mixed liquor.

The performance of a pilot-scale superoxygenation system was evaluated in clean water and mixed liquor. A mass balance was applied over the pilot-scale system to determine the overall oxygen mass transfer rate coefficient (KLa, h-1), the standard oxygen transfer rate (SOTR, kg O2 d-1), and the standard oxygen transfer efficiency (SOTE, %). Additionally, the alpha factor (α) was determined at a mixed liquor suspend solids (MLSS) concentration of approximately 5 g L-1. SOTEs of nearly 100% were obtained in clean water and mixed liquor. The results showed that at higher oxygen flowrates, higher transfer rates could be achieved; this however, at expenses of the transfer efficiency. As expected, lower transfer efficiencies were observed in mixed liquor compared to clean water. Alpha factors varied between 0.6 and 1.0. However, values of approximately 1.0 can be obtained in all cases by fine tuning the oxygen flowrate delivered to the system.

A pilot-scale microwave technology for sludge sanitization and drying.

Large volumes of sludge are produced from onsite sanitation systems in densely populated areas (e.g. slums and emergency settlements) and wastewater treatment facilities that contain high amounts of pathogens. There is a need for technological options which can effectively treat the rapidly accumulating sludge under these conditions. This study explored a pilot-scale microwave (MW) based reactor as a possible alternative for rapid sludge treatment. The reactor performance was examined by conducting a series of batch tests using centrifuged waste activated sludge (C-WAS), non-centrifuged waste activated sludge (WAS), faecal sludge (FS), and septic tank sludge (SS). Four kilograms of each sludge type were subjected to MW treatment at a power of 3.4kW for various time durations ranging from 30 to 240min. During the treatment the temperature change, bacteria inactivation (E. coli, coliforms, Staphylococcus aureus, and enterococcus faecalis) and sludge weight/volume reduction were measured. Calorific values (CV) of the dried sludge and the nutrient content (total nitrogen (TN) and total phosphorus (TP)) in both the dried sludge and the condensate were also determined. It was found that MW treatment was successful to achieve a complete bacterial inactivation and a sludge weight/volume reduction above 60%. Besides, the dried sludge and condensate had high energy (≥16MJ/kg) and nutrient contents (solids; TN≥28mg/g TS and TP≥15mg/g TS; condensate TN≥49mg/L TS and TP≥0.2mg/L), having the potential to be used as biofuel, soil conditioner, fertilizer, etc. The MW reactor can be applied for the rapid treatment of sludge in areas such as slums and emergency settlements.

Microwave treatment of faecal sludge from intensively used toilets in the slums of Nairobi, Kenya.

Toilet facilities in highly dense areas such as the slum and emergency settlements fill up rapidly; thus, requiring frequent emptying. Consequently, big quantities of fresh faecal sludge (FS) containing large amounts of pathogens are generated. Fast and efficient FS treatment technologies are therefore required for safe treatment and disposal of the FS in such conditions. This study explores the applicability of a microwave (MW) technology for the treatment of fresh FS obtained from urine-diverting dry toilets placed in slum settlements in Nairobi, Kenya. Two sample fractions containing 100 g and 200 g of FS were exposed to MW irradiation at three input MW power levels of 465, 1085 and 1550 W at different exposure times ranging from 0.5 to 14 min. The variation in the FS temperature, pathogen reduction via the destruction of E. coli and Ascaris lumbricoides eggs, and vol/wt reduction were measured during the MW treatment. It was demonstrated that the MW technology can rapidly and efficiently achieve complete reduction of E. coli and Ascaris lumbricoides eggs, and over 70% vol/wt reduction in the fresh FS. Furthermore, the successful evaluation of the MW technology under real field conditions demonstrated that MW irradiation can be applied for rapid treatment of fresh FS in situations such as urban slum and emergency conditions.

Evaluation of a microwave based reactor for the treatment of blackwater sludge.

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.

Metabolic Response of "Candidatus Accumulibacter Phosphatis" Clade II C to Changes in Influent P/C Ratio.

The objective of this study was to investigate the ability of a culture highly enriched with the polyphosphate-accumulating organism, "Candidatus Accumulibacter phosphatis" clade IIC, to adjust their metabolism to different phosphate availabilities. For this purpose the biomass was cultivated in a sequencing batch reactor with acetate and exposed to different phosphate/carbon influent ratios during six experimental phases. Activity tests were conducted to determine the anaerobic kinetic and stoichiometric parameters as well as the composition of the microbial community. Increasing influent phosphate concentrations led to increased poly-phosphate content and decreased glycogen content of the biomass. In response to higher biomass poly-phosphate content, the biomass showed higher specific phosphate release rates. Together with the phosphate release rates, acetate uptake rates also increased up to an optimal poly-phosphate/glycogen ratio of 0.3 P-mol/C-mol. At higher poly-phosphate/glycogen ratios (obtained at influent P/C ratios above 0.051 P-mol/C-mol), the acetate uptake rates started to decrease. The stoichiometry of the anaerobic conversions clearly demonstrated a metabolic shift from a glycogen dominated to a poly-phosphate dominated metabolism as the biomass poly-phosphate content increased. FISH and DGGE analyses confirmed that no significant changes occurred in the microbial community, suggesting that the changes in the biomass activity were due to different metabolic behavior, allowing the organisms to proliferate under conditions with fluctuating phosphate levels.

Temperature effects on glycogen accumulating organisms.

Glycogen accumulating organisms (GAO) compete for substrate with polyphosphate-accumulating organisms (PAO), which are the microorganisms responsible for the enhanced biological phosphorus removal (EBPR) in activated sludge wastewater treatment systems. This can lead to the deterioration of the EBPR process. In this paper, the long-term temperature effects on the anaerobic and aerobic stoichiometry and conversion rates on adapted enriched cultures of Competibacter (a known GAO) were evaluated from 10 to 40 degrees C. The anaerobic stoichiometry of Competibacter was constant from 15 to 35 degrees C, whereas the aerobic stoichiometry was insensitive to temperature changes from 10 to 30 degrees C. At 10 degrees C, likely due to the inhibition of the anaerobic conversions of Competibacter, a switch in the dominant bacterial population to an enriched Accumulibacter culture (a known PAO) was observed. At higher temperatures (35 and 40 degrees C), the aerobic processes limited the growth of Competibacter. Due to the inhibition or different steady-state (equilibrium) conditions reached at long-term by the metabolic conversions, the short- and long-term temperature dependencies of the anaerobic acetate uptake rate of Competibacter differed considerably between each other. Temperature coefficients for the various metabolic processes are derived, which can be used in activated sludge modeling. Like for PAO cultures: (i) the GAO metabolism appears oriented at restoring storage pools rather than fast microbial growth, and (ii) the aerobic growth rate of GAO seems to be a result of the difference between PHA consumption and PHA utilization for glycogen synthesis and maintenance. It appears that the proliferation of Competibacter in EBPR systems could be suppressed by adjusting the aerobic solids retention time while, aiming at obtaining highly enriched PAO cultures, EBPR lab-scale reactors could be operated at low temperature (e.g. 10 degrees C).

Modeling the PAO-GAO competition: effects of carbon source, pH and temperature.

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).

Temperature effects on the aerobic metabolism of glycogen-accumulating organisms.

Short-term temperature effects on the aerobic metabolism of glycogen-accumulating organisms (GAO) were investigated within a temperature range from 10 to 40 degrees C. Candidatus Competibacter Phosphatis, known GAO, were the dominant microorganisms in the enriched culture comprising 93 +/- 1% of total bacterial population as indicated by fluorescence in situ hybridization (FISH) analysis. Between 10 and 30 degrees C, the aerobic stoichiometry of GAO was insensitive to temperature changes. Around 30 degrees C, the optimal temperature for most of the aerobic kinetic rates was found. At temperatures higher than 30 degrees C, a decrease on the aerobic stoichiometric yields combined with an increase on the aerobic maintenance requirements were observed. An optimal overall temperature for both anaerobic and aerobic metabolisms of GAO appears to be found around 30 degrees C. Furthermore, within a temperature range (10-30 degrees C) that covers the operating temperature range of most of domestic wastewater treatment systems, GAOs aerobic kinetic rates exhibited a medium degree of dependency on temperature (theta = 1.046-1.090) comparable to that of phosphorus accumulating organisms (PAO). We conclude that GAO do not have metabolic advantages over PAO concerning the effects of temperature on their aerobic metabolism, and competitive advantages are due to anaerobic processes.

Factors affecting the microbial populations at full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants in The Netherlands.

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 (<0.1% as EUB), whereas Sphingomonas were not observed.

A practical method for quantification of phosphorus- and glycogen-accumulating organism populations in activated sludge systems.

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.

Short-term temperature effects on the anaerobic metabolism of glycogen accumulating organisms.

Proliferation of glycogen accumulating organisms (GAO) has been identified as a potential cause of enhanced biological phosphorus removal (EBPR) failure in wastewater treatment plants (WWTP). GAO compete for substrate with polyphosphate accumulating organisms (PAO) that are the microorganisms responsible for the phosphorus removal process. In the present article, the effects of temperature on the anaerobic metabolism of GAO were studied in a broad temperature range (from 10 to 40 degrees C). Additionally, maximum acetate uptake rate of PAO, between 20 and 40 degrees C, was also evaluated. It was found that GAO had clear advantages over PAO for substrate uptake at temperatures higher than 20 degrees C. Below 20 degrees C, maximum acetate uptake rates of both microorganisms were similar. However, lower maintenance requirements at temperature lower than 30 degrees C give PAO metabolic advantages in the PAO-GAO competition. Consequently, PAO could be considered to be psychrophilic microorganisms while GAO appear to be mesophilic. These findings contribute to understand the observed stability of the EBPR process in WWTP operated under cold weather conditions. They may also explain the proliferation of GAO in WWTP and thus, EBPR instability, observed in hot climate regions or when treating warm industrial effluents. It is suggested to take into account the observed temperature dependencies of PAO and GAO in order to extend the applicability of current activated sludge models to a wider temperature range.