Enhanced Biological Phosphorus Removal Performance Research Articles

The impact of reduced sulfur compounds on aerobic granular sludge formation and biological phosphorus removal

Biological sulfur conversion combined with phosphorus removal represents an innovative biotechnology for activated sludge (AS). However, its effect on the formation of aerobic granular sludge (AGS) remains unclear. This study aimed at determining the impact of sulfide and thiosulfate on the performance of enhanced biological phosphorus removal (EBPR) and on AGS formation. Using two lab-scale sequencing batch reactors (SBR), we compared EBPR performance, the degree of granulation, and the enriched microbial community with sulfide (SBR-S, 20 mg S/L) or thiosulfate (SBR-T, 140 mg S/L) as primary sulfur source. Results indicate that both reactors achieved comparable EBPR performance, with a phosphorus removal rate of 98% in SBR-T and 99% in SBR-S. SBR-T resulted in compact granules with fast settling properties, while SBR-S resulted in dense floccular sludge, referred to as “densified sludge”. Both systems enriched a comparable microbial community with shared dominant genera, including Ca. Competibacter, Thiothrix, Ca. Accumulibacter and Dechloromonas. The higher abundance of Thiothrix unzii throughout the operating time in SBR-T, reaching 19.4%, is believed to have enhanced the granulation process. This study provides important insight into the role of reduced sulfur compounds in sludge densification and EBPR performance, relevant to sulfur-rich wastewaters such as those from industrial sources.

Enhanced biological phosphorus removal from wastewater and dynamics of microbial population in dual PAOs symbiosis system

Low level of volatile fatty acids (VFAs) is a key factor limiting the performance of enhanced biological phosphorus removal (EBPR) by traditional polyphosphate accumulating organisms (PAOs). A newly discovered PAOs capable of fermenting macromolecular organic matter into short-chain VFAs could solve the problem. However, the roles of traditional and fermented PAOs in wastewater treatment processes and the collaboration between functional microorganisms remain unknown. In this study, a sequencing batch reactor (SBR) was operated with alternating anaerobic oxic (An/O) mode to investigate the effect of mixed carbon source ratios, namely, acetate (NaAc):casein hydrolysate (Cas aa) = 4:0, 3:1, 1:1, 1:3 and 0:4, on EBPR performance and microbial community structure. The results showed that the phosphorus removal efficiency remained stable as NaAc was the sole carbon source. When NaAc:Cas aa varied from 3:1 to 1:3, the recovery of system performance took less time compared to the last Run, implying an adaptation of PAOs to Cas aa. Moreover, the anaerobic phosphorus release time was found to increase with the increasing amount of VFAs from Cas aa degradation, indicating the enrichment of fermented bacteria and the utilization of VFAs by traditional PAOs. Increasing the dose of Cas aa also generated more NH4+-N, owing to the reaction of the groups in the amino acids with water, causing a drastic decrease of ammonium removal efficiency from 88.9 % to 15.2 %. The ideal process performance implied that EBPR by dual PAOs synergistic symbiosis occurred in the system with NaAc:Cas aa = 1:1. Meanwhile, the results of 16S ribosomal RNA (16S rRNA) genes-Illumina MiSeq high-throughput sequencing showed that the abundance of traditional PAOs decreased from 22.4 % (4:0) to 11.6 % (1:1) and to 4.9 % (0:4), but fermented PAOs increased from 9.9 % to 19.5 %, then decreased to 17.3 %. Dual PAOs synergistic symbiosis could be of great potential for EBPR in treating low VFAs wastewater.

Toward Integrating EBPR and the Short-Cut Nitrogen Removal Process in a One-Stage System for Treating High-Strength Wastewater.

Enhanced biological phosphorus removal (EBPR) is an economical and sustainable process for phosphorus removal from wastewater. Despite the widespread application of EBPR for low-strength domestic wastewater treatment, limited investigations have been conducted to apply EBPR to the high-strength wastewaters, particularly, the integration of EBPR and the short-cut nitrogen removal process in the one-stage system remains challenging. Herein, we reported a novel proof-of-concept demonstration of integrating EBPR and nitritation (oxidation of ammonium to nitrite) in a one-stage sequencing batch reactor to achieve simultaneous high-strength phosphorus and short-cut nitrogen removal. Excellent EBPR performance of effluent 0.8 ± 1.0 mg P/L and >99% removal efficiency was achieved fed with synthetic high-strength phosphorus wastewater. Long-term sludge acclimation proved that the dominant polyphosphate accumulating organisms (PAOs), Candidatus Accumulibacter, could evolve to a specific subtype that can tolerate the nitrite inhibition as revealed by operational taxonomic unit (OTU)-based oligotyping analysis. The EBPR kinetic and stoichiometric evaluations combined with the amplicon sequencing proved that the Candidatus Competibacter, as the dominant glycogen accumulating organisms (GAOs), could well coexist with PAOs (15.3-24.9% and 14.2-33.1%, respectively) and did not deteriorate the EBPR performance. The nitrification activity assessment, amplicon sequencing, and functional-based gene marker quantification verified that the unexpected nitrite accumulation (10.7-21.0 mg N/L) in the high-strength EBPR system was likely caused by the nitritation process, in which the nitrite-oxidizing bacteria (NOB) were successfully out-selected (<0.1% relative abundance). We hypothesized that the introduction of the anaerobic phase with high VFA concentrations could be the potential selection force for achieving nitritation based on the literature review and our preliminary batch tests. This study sheds light on developing a new feasible technical route for integrating EBPR with short-cut nitrogen removal for efficient high-strength wastewater treatment.

The impact of pH on the anaerobic and aerobic metabolism of Tetrasphaera-enriched polyphosphate accumulating organisms

Members of the genus Tetrasphaera are putative polyphosphate accumulating organisms (PAOs) that have been found in greater abundance than Accumulibacter in many full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants worldwide. Nevertheless, previous studies on the effect of environmental conditions, such as pH, on the performance of EBPR have focused mainly on the response of Accumulibacter to pH changes. This study examines the impact of pH on a Tetrasphaera PAO enriched culture, over a pH range from 6.0 to 8.0 under both anaerobic and aerobic conditions, to assess its impact on the stoichiometry and kinetics of Tetrasphaera metabolism. It was discovered that the rates of phosphorus (P) uptake and P release increased with an increase of pH within the tested range, while PHA production, glycogen consumption and substrate uptake rate were less sensitive to pH changes. The results suggest that Tetrasphaera PAOs display kinetic advantages at high pH levels, which is consistent with what has been observed previously for Accumulibacter PAOs. The results of this study show that pH has a substantial impact on the P release and uptake kinetics of PAOs, where the P release rate was >3 times higher and the P uptake rate was >2 times higher at pH 8.0 vs pH 6.0, respectively. Process operational strategies promoting both Tetrasphaera and Accumulibacter activity at high pH do not conflict with each other, but lead to a potentially synergistic impact that can benefit EBPR performance.

Enhancing resource recovery via cranberry syrup waste at the Wisconsin Rapids WRRF: An experimental and modeling study

The Wisconsin Rapids Wastewater Treatment Plant (WRWWTP) is faced with a more stringent effluent phosphorus requirement that will drive capital investment between 2020 and 2025. The facility will need to achieve a monthly average value of 0.36 mg L−1 of total phosphorus (TP). While the facility has sufficient influent carbon to drive a conventional enhanced biological phosphorus removal (EBPR) configuration, the existing infrastructure makes the addition of influent selector zones cost prohibitive. Underutilized aeration basin capacity was repurposed for testing return activated sludge (RAS) fermentation. The WRWWTP began pilot testing of RAS fermentation in April 2021. The facility moved through a series of operational setpoints to optimize phosphorus removal in a sidestream RAS (SSR) configuration, including RAS diversion, decrease of DO in aeration basins and chemical dosing shutoff. One of the key implementations was the addition of cranberry syrup waste to provide additional carbon for RAS fermentation, converting the process to a SSR plus carbon (SSRC) configuration. By the end of the testing period, effluent total phosphorus was averaging less than 0.4 mg L−1 with no chemical addition. A model was developed in the SUMO platform and was used to capture orthophosphate trends during the testing period. The model investigated microbial population dynamics and found that the operational changes including RAS diversion, chemical dosing shutoff and cranberry syrup waste addition impacted the enrichment of phosphorus accumulating organisms (PAO). After performing a sensitivity analysis on hydrolysis parameters, the predicted hydrolysis rate around 1.8–1.9 mg COD g VSS−1 hr−1 was found to match the batch rate testing data. This is the first study where cranberry syrup waste was used to successfully enhance EBPR performance, resulting in 90% TP removal. While further research is needed regarding the composition of the waste matrix and the microbial community composition, this expands the routes for resource recovery in the field of wastewater treatment.

Phosphorus removal performance, intracellular metabolites and clade-level community structure of Tetrasphaera-dominated polyphosphate accumulating organisms at different temperatures

Tetrasphaera are polyphosphate accumulating organisms (PAOs) that play an important role in enhanced biological phosphorus removal (EBPR) from wastewater. The effect of a wide range of temperature changes (1–30 °C) on phosphorus removal, metabolism and clade-level community structure of Tetrasphaera-dominated PAOs was investigated. At 10 °C, the bioactivities of Tetrasphaera-dominated communities were obviously inhibited and the EBPR efficiency was only 73 %. Yet at 20–30 °C, EBPR efficiency reached 99 % and the relative abundance of Tetrasphaera was up to 90 %. The temperature variation changed the community distribution of Tetrasphaera clades, which was possibly a main reason for EBPR performance. Amino acids and PHA with different contents were intracellular metabolite of Tetrasphaera-dominated communities during phosphorus release and uptake at different temperatures. Moreover, Tetrasphaera fermented protein and amino acids and released VFAs. The outcomes suggested the great potential of Tetrasphaera-PAOs in the treatment of wastewater with varying temperatures and limited carbon sources.

Biological phosphorus removal and its microbial community in a modified full-scale activated sludge system under dry and wet weather dynamics

Enhanced biological phosphorus removal (EBPR) performance and microbial community dynamics during dry and wet-weather conditions of a full-scale treatment plant was evaluated by converting a section of activated sludge basins using low-cost operational modifications into an anoxic/anaerobic zone to promote EBPR. Two trains of the activated sludge system at the Des Moines, Iowa Metropolitan Wastewater Reclamation Facility were used for the study with one train modified for EBPR, and the other remained as nitrification-only for comparison.In addition to measuring the modification effectiveness for phosphorus removal, performance was compared during dry and wet weather conditions over the course of two summer seasons to improve understanding of wet and dry weather dynamics for EBPR. DNA sequencing and qPCR tests were conducted to develop an understanding of microbial population changes between control and modified basins and wet and dry weather conditions.Basin hydraulic retention times varied from 2.6 to 12.7 hours with an average of 8.9 hours. EBPR activity was successfully established in the modified basins with average phosphorus content of the return activated sludge 0.032 ± 0.002 compared to 0.016 ± 0.001 mg TP/mg TSS (95% confidence) in the control basins. Phosphorus removal was significantly decreased by prolonged wet weather conditions, particularly in year two of the study, however the modified basin maximum removal of 96% and average of 43.7 ± 5.3% remained significantly higher than the maximum of 46% and average 12.6 ± 2.4% removal in the control basins.DNA sequencing showed a significant increase in relative abundance of phyla Chloroflexi, Nitrospirae, and Verrucomicrobia in the modified basins, but no correlation to EBPR performance. qPCR indicated significant increase in relative quantity of Accumulibacter, but not for Actinetobacter-like phosphorus accumulating organisms (PAOs), which includes the PAO Tetrasphaera. Significant abundance of some Accumulibacter clades found within the modified basins was contrary to previous literature which focused on small-scale and batch studies. A higher than expected dominance of clade I and increased relative quantities of clades IIB and IIC during extended wet weather was observed which may have contributed to rapid recovery of phosphorus removal when dry weather resumed. The abundance of PAOs did not significantly correlate with changes in phosphorous removal performance, contrary to reports from previous small-scale and batch studies.

Long-term exposure to zinc oxide nanoparticles improves PAOs function in enhanced biological phosphorus removal

ABSTRACT As the most widely applied process for biological phosphorus removal, enhanced biological phosphorus removal (EBPR) relies on phosphorus accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), whose function is crucial for the removal of phosphorus. In this study, the effect of zinc oxide nanoparticles (ZnO NPs, 0–50 mg/L) on EBPR performance was investigated in both long-term reactors and batch experiments. It was found that the performance of biological phosphorus removal was recovered from 0% (day 0) to >99% (day 70) after long-term exposure of ZnO NPs (50 mg/L). Further studies revealed that ZnO NPs treatment caused no significant effects on the morphology and settleability of activated sludge, but enhanced the release and uptake of phosphorus as well as the transformations of polyhydroxyalkanoates and glycogen in activated sludge, which suggested that PAOs were re-activated during long-term exposure to ZnO NPs. Fluorescence in-situ hybridization (FISH) analysis showed that the relative abundance of PAOs was increased after long-term exposure. Meanwhile, the enzymatic activities of PPX and PPK were also enhanced. These results indicated that compared with short-term exposure, long-term exposure to ZnO NPs favours PAOs function and thus led to the recovery of biological phosphorus removal.

Intracellular polyphosphate length characterization in polyphosphate accumulating microorganisms (PAOs): Implications in PAO phenotypic diversity and enhanced biological phosphorus removal performance

Polyphosphate (polyP) accumulating organisms (PAOs) are the key agent to perform enhanced biological phosphorus removal (EBPR) activity, and intracellular polyP plays a key role in this process. Potential associations between EBPR performance and the polyP structure have been suggested, but are yet to be extensively investigated, mainly due to the lack of established methods for polyP characterization in the EBPR system. In this study, we explored and demonstrated that single-cell Raman spectroscopy (SCRS) can be employed for characterizing intracellular polyPs of PAOs in complex environmental samples such as EBPR systems. The results, for the first time, revealed distinct distribution patterns of polyP length (as Raman peak position) in PAOs in lab-scale EBPR reactors that were dominated with different PAO types, as well as among different full-scale EBPR systems with varying configurations. Furthermore, SCRS revealed distinctive polyP composition/features among PAO phenotypic sub-groups, which are likely associated with phylogenetic and/or phenotypic diversity in EBPR communities, highlighting the possible resolving power of SCRS at the microdiversity level. To validate the observed polyP length variations via SCRS, we also performed and compared bulk polyP length characteristics in EBPR biomass using conventional polyacrylamide gel electrophoresis (PAGE) and solution 31P nuclear magnetic resonance (31P-NMR) methods. The results are consistent with the SCRS findings and confirmed the variations in the polyP lengths among different EBPR systems. Compared to conventional methods, SCRS exhibited advantages as compared to conventional methods, including the ability to characterize in situ the intracellular polyPs at subcellular resolution in a label-free and non-destructive way, and the capability to capture subtle and detailed biochemical fingerprints of cells for phenotypic classification. SCRS also has recognized limitations in comparison with 31P-NMR and PAGE, such as the inability to quantitatively detect the average polyP chain length and its distribution. The results provided initial evidence for the potential of SCRS-enabled polyP characterization as an alternative and complementary microbial community phenotyping method to facilitate the phenotype-function (performance) relationship deduction in EBPR systems.

Tetrasphaera, rather than Candidatus Accumulibacter as performance indicator of free ammonia inhibition during the enhanced biological phosphorus removal processes

Free ammonia (FA) has been proposed to inhibit phosphorus (P) removal efficiency and reconfigures the microbial community in the enhanced biological phosphorus removal (EBPR) systems. However, in response to FA exposure, the microbial kinetics and the functional bacteria belonging to polyphosphate accumulating organisms (PAOs) i.e., Tetrasphaera and Candidatus Accumulibacter within the mixed communities are generally ignored. This work was conducted to characterize the effects of FA on the biological processes of P-release and its uptake, and the virtual role of PAOs in EBPR systems. Experimental data demonstrated that the FA strongly inhibited the P-release and uptake rates which was described well by Hellinga and Vadivelu models, and the determined R2 in the anaerobic and aerobic period were about 0.91 and 0.96. Moreover, the abundance of key microbial communities differed significantly with various FA concentrations. Structural equation model (SEM) revealed that FA was positively associated with Ca. Accumulibacter (path coeff. = 0.30; P = 0.13) and Ca. Competibacter (path coeff. = 0.43; P < 0.05), while it was negatively associated with Tetrasphaera (path coeff. = − 0.45; P < 0.001). Furthermore, Tetrasphaera was found to strongly and positively drive the rates of P-release and uptake, and was confirmed as the only virtual biomarker for indicating the response of EBPR performance towards FA inhibition. Two hypothetical mechanisms indicated that Ca. Accumulibacter under higher FA concentration could not synthesize PHA to support the EBPR process. In addition, the release of P was hindered due to the FA toxicity on the urea cycle of Tetrasphaera microbial cells. This work gives comprehensive and deep insight into the effect of FA on the EBPR system and provides a useful guide for process optimization.

Evaluation of PAO adaptability to oxygen concentration change: Development of stable EBPR under stepwise low-aeration adaptation

Recent research has shown the adaptability of Biological Nutrient Removal (BNR) systems to very low level dissolved oxygen (DO) concentration, mainly focusing in the nitrification ability that maintains the nitrogen oxidation process even at very low DO levels. Although step-wise aeration decrease on Enhanced Biological Phosphorus Removal (EBPR) is not fully comprehended. This study investigated the effect of reaching micro-aeration with adaptation strategies on EBPR performance. A step-wise oxygen concentration decrease, arriving at an average aeration level of 0.4 mgO2/L was evaluated, with an outcome of approximately 90 % phosphorus removal efficiency. Compared with different aeration modes, the highest phosphorus (P)-removal efficiency, P-release and lowest effluent phosphorus was achieved in gradual DO decrease strategy. On the other hand, an instant decrease in aeration from stable EBPR process from 2 mgO2/L to 0.4 mgO2/L adversely impacted P-removal by decreasing the efficiency to average 60 % and deteriorating the phosphorus removing microbial consortium. Comparison of results between instant and gradual DO-decrease, indicated the sensitivity of microorganisms to aeration. Microbial adaptation to decreased oxygen availability is crucial to reach high process performance. This study proposes, a potential aeration mode, which contributes in reduction of energy consumption in BNR systems through wastewater treatment.

Accumulibacter diversity at the sub-clade level impacts enhanced biological phosphorus removal performance

Accumulibacter is a well-known group of organisms, typically considered to be polyphosphate accumulating organisms (PAOs), but potentially capable of glycogen accumulating organism (GAO) metabolism under limiting influent phosphate levels. Metabolic features of Accumulibacter are typically linked to its phylogenetic identity at the Type or clade level, though it is unclear the extent to which Accumulibacter diversity can correlate with its capacity to perform P removal. This paper investigates the fine-scale diversity of Accumulibacter and its link with enhanced biological phosphorus removal (EBPR) performance under various operating conditions, to understand the conditions and community structure leading to successful and unsuccessful EBPR operation. For this purpose, the organic carbon feeding rate and total organic carbon concentration were varied during three distinct operational periods, where influent phosphate was never limiting. Accumulibacter was always the dominant microbial group (>80% of all bacteria according to quantitative fluorescence in situ hybridisation - FISH) and low levels of Competibacter and other GAOs were consistently observed (<15% of all bacteria). Steady state was achieved in each of the three periods, with average phosphorus removal levels of 36%, 99% and >99%, respectively. Experimentally determined stoichiometric activity supported the expression of a mixed PAO/GAO metabolism in the first steady state period and the typical PAO metabolism in the other two steady state periods. FISH quantification and amplicon sequencing of the polyphosphate kinase (ppk1) functional gene indicated that Accumulibacter clade IIC was selected in the first steady state period, which shifted to clade IA after decreasing the carbon feeding rate in steady state period 2, and finally shifted back to clade IIC in the third steady state period. Fine-resolution Ppk-based phylogenetic analysis revealed three different clusters within Accumulibacter clade IIC, where clusters IICii and IICiii were linked to poor EBPR performance in period 1, and cluster IICi was linked to good EBPR performance in period 3. This study shows that the deterioration of EBPR processes through GAO activity at non-limiting P concentrations can be linked to organisms that are typically classified as PAOs, not only to known GAOs such as Competibacter. Intra-clade phylogenetic diversity within Accumulibacter showed that some clusters actually behave similarly to GAOs even without influent phosphate limitation. This study highlights the need to closely re-examine traditional interpretations regarding the link between the microbial community composition and identity with the performance and metabolism of EBPR systems.

Effects of anaerobic HRT and VFA loading on the kinetics and stoichiometry of enhanced biological phosphorus removal

Enhanced biological phosphorus removal (EBPR) can recover significant quantities of wastewater phosphorus. However, this resource recovery process realizes limited use largely due to process stability concerns. Research evaluated effects of anaerobic HRT (τAN ) and VFA concentration - critical operational parameters that can be externally controlled - on EBPR performance. Evaluated alone, τAN (1-4 hours) exhibited no statistical effect on effluent phosphorus. However, PHA increased with VFA loading and biomass accumulated more phosphorus. Regarding resiliency, under increasing VFA loads PAOs hydrolyzed more phosphorus to uptake/catabolize VFAs; moreover, PHA synthesis normalized to VFA loading increased with τAN , suggesting fermentation. Kinetically, PAOs exhibited Monod-like relationships for qPHAAN and qVFAAN as a function of anaerobic P release; additionally, qPAE exhibited a Monod-like relationship with end-anaerobic PHA concentration. A culminating analysis affirmed the relationship between enhanced aerobic P uptake, and net P removal, with a parameter (Phosphorus Removal Propensity Factor) that combines influent VFA concentration with τAN .

P.080 A translational Homer 1a-based network approach: imaging how haloperidol modulates glutamate system functional connectivity

The dynamic response mechanism of Candidatus Accumulibacter clades to environmental factors in enhanced biological phosphorus removal (EBPR) was unclear. This study investigated the relationship between the transcriptional responses of Candidatus Accumulibacter clades and environmental dynamics. Results suggested that Candidatus Accumulibacter clade IIA only responded in initial 20 and 30 min of P-release and P-uptake stage, respectively, and was also the first clade to stop responding among the six Candidatus Accumulibacter clades. Clade IIC and IID responded at rising stage of P-release and P-uptake rate. Clade IA and IIB responded at decreasing stage of P-release and P-uptake rate. The transcriptional response duration of clade IIF was the longest, which constantly responded throughout anaerobic, anoxic and oxic phase. The transcriptional responses of Candidatus Accumulibacter clades to environmental dynamics revealed the microorganisms actually working in P-release and P-uptake, and gave a new insight into the transcriptional responses related to the EBPR performance.

Defluviicoccus vanus Glycogen-Accumulating Organisms (DvGAOs) Are Less Competitive Than Polyphosphate-Accumulating Organisms (PAOs) at High Temperature

Glycogen-accumulating organisms (GAOs) have been previously observed to be more competitive in anaerobic volatile fatty acid (VFA) uptake as compared to polyphosphate-accumulating organisms (PAOs) at high temperatures in enhanced biological phosphorus removal (EBPR) systems. However, previous evidence regarding the impact of temperature on GAOs has exclusively focused on Competibacter. Defluviicoccus vanus GAOs (DvGAOs) make up another very important GAO group and have been found in EBPR plants; however, their competitiveness at high temperature for anaerobic VFA uptake was previously unknown. The rate of uptake of VFAs by both DvGAOs reactors enriched with acetate or propionate was much lower than those of Candidatus Accumulibacter phosphatis PAOs operated under similar conditions, showing that PAOs displayed a competitive advantage at high temperature, unlike previous observations with Competibacter. This study provides new insight into our understanding of GAOs in EBPR processes, where the impact of temperature on PAO-GAO competition is highly dependent on the phylogenetic identity of the GAO present in the system. New strategies for promoting EBPR performance at high temperatures are proposed on the basis of the results of this work.

Long-term simulation of a full-scale EBPR plant with a novel metabolic-ASM model and its use as a diagnostic tool

This study evaluates the predictive capacity of the META-ASM model, a new integrated metabolic activated sludge model, in describing the long-term performance of a full-scale enhanced biological phosphorus removal (EBPR) system that suffers from inconsistent performance. In order to elucidate the causes of EBPR upsets and troubleshoot the process accordingly, the META-ASM model was tested as an operational diagnostic tool in a 1336-day long-term dynamic simulation, while its performance was compared with the ASM-inCTRL model, a version based on the Barker & Dold model. Overall, the predictions obtained with the META-ASM without changing default parameters were more reliable and effective at describing the active biomass of polyphosphate accumulating organisms (PAOs) and the dynamics of their storage polymers. The primary causes of the EBPR upsets were the high aerobic hydraulic retention times (HRTs) and low organic loading rates (OLRs) of the plant, which led to periods of starvation. The impact of these factors on EBPR performance were only identified with the META-ASM model. Furthermore, the first signs of process upsets were predicted by variations in the aerobic PAO maintenance rates, suggesting that the META-ASM model has potential to provide an early warning of process upset. The simulation of a new viable operational strategy indicated that troubleshooting the process could be achieved by reducing the aerated volume by switching off air in the first half of the aeration tank. In this new strategy, the META-ASM model predicted a simultaneous improvement in the biological phosphorus (P) and nitrogen (N) removal due to the enhancement of the hydrolysis and fermentation of the mixed liquor sludge in the new unaerated zone, which increased the availability of volatile fatty acids (VFAs) for PAOs. This study demonstrates that the META-ASM model is a powerful operational diagnostic tool for EBPR systems, capable of predicting and mitigating upsets, optimising performance and evaluating new process designs.

Transcriptional responses of Candidatus Accumulibacter clades to environmental dynamics in enhanced biological phosphorus removal

The dynamic response mechanism of Candidatus Accumulibacter clades to environmental factors in enhanced biological phosphorus removal (EBPR) was unclear. This study investigated the relationship between the transcriptional responses of Candidatus Accumulibacter clades and environmental dynamics. Results suggested that Candidatus Accumulibacter clade IIA only responded in initial 20 and 30 min of P-release and P-uptake stage, respectively, and was also the first clade to stop responding among the six Candidatus Accumulibacter clades. Clade IIC and IID responded at rising stage of P-release and P-uptake rate. Clade IA and IIB responded at decreasing stage of P-release and P-uptake rate. The transcriptional response duration of clade IIF was the longest, which constantly responded throughout anaerobic, anoxic and oxic phase. The transcriptional responses of Candidatus Accumulibacter clades to environmental dynamics revealed the microorganisms actually working in P-release and P-uptake, and gave a new insight into the transcriptional responses related to the EBPR performance.

New insights in the competition of polyphosphate-accumulating organisms and glycogen-accumulating organisms under glycogen accumulating metabolism with trace Poly-P using flow cytometry

In enhanced biological phosphorus removal (EBPR) from wastewater, the metabolism pathway of polyphosphate accumulating organisms (PAOs) converted from polyphosphate accumulating metabolism (PAM) to glycogen accumulating metabolism (GAM) under a high ratio of carbon to phosphorus (C/P). This study investigated the competition mechanism between PAOs and glycogen accumulating organisms (GAOs) under GAM pathway. Low acetic acid (HAC) concentration made the GAM of PAOs outcompete GAOs in high C/P ratio (100:1), and the effect of influent acetate concentration on GAOs was stronger than PAOs. “Candidatus Accumulibacter” clades IIC and IID were the main dominant PAOs, and Thiothrix caldifontis (potential PAOs) was discovered and significantly increased from 0.53% to 5.75%. Flow cytometry results indicated that PAOs had higher specific surface area and lower nucleic acid content than GAOs, and high C/P promoted GAM rather than GAOs growth, which maybe important reasons causing PAOs dominant growth. PAOs dominance under GAM ensured stable performance of EBPR when C/P ratio in real wastewater increased.

A novel micro-ferrous dosing strategy for enhancing biological phosphorus removal from municipal wastewater

Ferrous salts have been widely used to enhance phosphorus removal in full-scale wastewater treatment plants, with an average dosage of 0.24–0.35 mM. However, such high dosage inevitably caused serious concerns on operation, potential biological toxicity and excessive sludge production. Thus, this study investigated the effect of micro-dosing of ferrous salt at the level of 0.02 mM on enhanced biological phosphorus removal (EBPR) in sequencing batch reactors. Results showed that micro-dosing of ferrous salt enhanced the overall performance, with average COD, TN and TP removal of more than 4.2%, 2.0% and 5.8%, respectively. In addition, the sequencing analysis further revealed that micro-ferrous dosing could significantly improve the diversity and richness of the microbial community (p < 0.05), whereas the regular dosing of ferrous salts (0.25 mM) negatively impacted on the EBPR performance. It was found that the abundances of phosphorus accumulating organisms (PAOs) in R2 (micro-dosing) were nearly 1.5-fold and 2-fold higher than those in R1 (control) and R3 (regular dosing). The contributions of biological and chemical pathways towards the observed phosphorus removal were also determined according to the phosphorus releasing rate. For micro-dosage and regular dosage of ferrous salts, phosphorus removal mainly relied on biological phosphorus removal and chemical phosphorus removal, respectively. It appears from this this study that the micro-ferrous dosing strategy is practically feasible and economically viable for enhanced phosphorus removal from municipal wastewater.

Investigation of anaerobic side-stream phosphorus recovery and its effect on the performance of mainstream EBPR subjected to low-consumption.

In this study, phosphate-rich supernatant at the end of anaerobic phase was extracted by a certain side-stream ratio for chemical precipitation to investigate the optimal conditions for phosphorus recovery. The effect of side-stream reaction on the performance of the mainstream enhanced biological phosphorus removal (EBPR) system was also explored. The experiment was carried out in a sequencing batch reactor (SBR) operated in an alternating anaerobic/aerobic mode with dissolved oxygen controlled at 1.0 mg · L-1. The results showed that the optimum magnesium source,temperature, stirring speed and reaction equilibrium time for side-stream phosphorus recovery were: MgCl2 · 6H2O, 25 °C, 150 rpm and 20 min, respectively. It was also observed that the average phosphorus removal efficiency of the mainstream system maintained as high as 90.7% during the side-stream extraction period despite insufficient time for phosphate uptake under limited dissolved oxygen condition and phosphate deprivation of polyphosphate-accumulating organisms (PAOs). Besides, the sludge settling performance of the mainstream EBPR system decreased with no sludge loss. Afterwards, phosphorus removal and sludge settling performance were restored with dismissing side-stream phosphorus recovery. This study suggested that side-stream extraction of anaerobic supernatant from a mainstream EBPR subjected to low dissolved oxygen conditions for chemical phosphorus recovery was feasible and environmentally friendly.