Denitrifying Phosphorus Accumulating Organisms Research Articles

Balancing denitrifying phosphorus-accumulating organisms and denitrifying glycogen-accumulating organisms for advanced nitrogen and phosphorus removal from municipal wastewater

Given the carbon limitation of municipal wastewater, the balance of biological nitrogen and phosphorus removal remains a challenging task. In this study, an anaerobic-anoxic–oxic combining with biological contact oxidation (A2/O-BCO) system treating real municipal wastewater was operated for 205 days, and COD-to-PO43--P ratio was confirmed as the key parameter for balancing denitrifying phosphorus-accumulating organisms (DPAOs) and denitrifying glycogen-accumulating organisms (DGAOs) to enhance N and P removal. When DPAOs dominated in nutrients removal, the increase in COD/P from 17.1 to 38.1 caused the deterioration in nitrogen removal performance decreasing to 71.8 %. As COD/P ratio decreased from 81.3 to 46.8, Ca.Competibacter proliferated from 3.11 % to 6.00 %, contributing to 58.9 % of nitrogen removal. The nitrogen and phosphorus removal efficiency reached up to 79.3 % and 95.2 %. Overall, establishing DGAOs-DPAOs balance by strengthening the effect of DGAOs could enhance the nutrients removal performance and accordingly improve the stability and efficiency of the system.

Mathematical modeling of the dynamic effect of denitrifying glycogen-accumulating organisms on nitrous oxide production during denitrifying phosphorus removal

The large amount of intermediate nitrous oxide (N2O) production from denitrifying phosphorus removal (DPR) increases the carbon footprint of wastewater treatment. However, there is a lack of detailed understanding of the interrelationships between denitrifying polyphosphate-accumulating organisms (DPAOs) and denitrifying glycogen-accumulating organisms (DGAOs) on N2O production during DPR. In this work, a mathematical model was developed for the first time to describe dynamic N2O production in the DPR system coexisting DPAOs and DGAOs. The model took into account a four-step subsequent denitrification of nitrate, nitrite, nitric oxide, and N2O. The validity of model was fully tested by comparing simulation studies with experimental data from three independent reports on DPR, which satisfactorily described the dynamics of N2O production, nitrogen oxide reduction, phosphate release and uptake, and intracellular polymers turnover. The validated model could clarify the source and pathways of N2O production. Subsequently, the combined effects of key operational conditions on the overall N2O production, DPAOs and DGAOs competition, and nutrients removal efficiency were investigated by model simulation. Simulation results showed higher polyhydroxyalkanoate storage rate of DGAOs than that of DPAOs during anaerobic stage and the preference of DGAOs for nitrate electron acceptor during anoxic stage. DGAOs dominated the growth competition over DPAOs at low COD (<150 mg/L) and high nitrate (>35 mg/l) conditions, leading to the anoxic storage of glycogen by DGAOs as the main pathway of N2O production. In addition, both N2O production and generation pathways in the system possessed greater variability compared to single DGAOs or DPAOs system, further indicating the necessity of this model.

Study on the mechanisms of enhanced biological nitrogen and phosphorus removal by denitrifying phosphorus removal in a Micro-pressure swirl reactor

To reveal the mechanisms of enhanced biological nitrogen and phosphorus removal by denitrifying phosphorus removal in a Micro-pressure swirl reactor (MPSR), this study used a MPSR to treat municipal wastewater and enriched denitrifying phosphate accumulating organisms (DPAOs) by using its alternating anaerobic-anoxic-aerobic environment. The coupling of denitrification phosphorus removal (DPR) and simultaneous nitrification endogenous denitrification phosphorus removal (SNEDPR) was achieved in MPSR, and the average removal rates of COD, NH4+-N, TN and TP were 91.57%, 98.51%, 85.88%, 96.08% respectively. The results of the batch experiments showed that DPAOs activity in the low dissolved oxygen (DO) and high DO zones were 70.5% and 74.3%. The results of intracellular carbon source conversion patterns, microbial assays and functional gene prediction indicated that Flavobacterium and Dechloromonas dominated the DPR process in the low DO zone. Based on these findings, nutrient removal pathways within the MPSR were integrated.

La–Fe magnetic bentonite stimulated denitrifying phosphorus removal from low C/N wastewater in the A2/O process: Performance, microbial community, and potential mechanism

A novel process combined A2/O bioreactor with a millimeter-sized magnetic lanthanum-modified bentonite (La–Fe-MB) granules material (terms as A2/O–La–Fe-MB) was proposed for highly efficient phosphorus removal from municipal sewage. A series of experiments were carried out to explore the feasibility and suitability of using La–Fe-MB as a novel adsorbent. The performance of nutrient removal and microbial composition, and any adverse effect brought by dosing La–Fe-MB are of interests. The long-term operation (240 days) suggested that dosing La–Fe-MB in the A2/O bioreactor achieved a highly-efficient TP removal with average effluent TP concentrations for 0.22 mg/L by enhancing denitrifying phosphorus removal (DPR) and chemical precipitation. Based on the mass balance equations, La–Fe-MB contributed to ∼39.7% of TP removal via adsorption/precipitation in the anaerobic zone, 43.6% of TP was removed through anoxic DPR in the anoxic zone, and post-aerobic zone further reduced ∼14.4% of phosphorus. Moreover, an interesting synergic effect between the La–Fe-MB and functional bacteria was found in the combined process. La–Fe-MB addition not only have less effect on the removal of organic, nitrogen species, and functional bacteria (AOB, NOB, etc.,), but significantly facilitated the enrichment of denitrifying phosphorus accumulating organisms (DPAOs) via releasing traces of Fe (Fe(II)/Fe(III)) in bioreactor. Besides, DPAOs (1.7%), glycogen accumulating organisms (GAOs) (0.031%) and P-accumulating organisms (PAOs) (0.41%) co-existed in this combined system, potentially resulting in simultaneous denitrifying dephosphatation and aerobic phosphorus absorbing. The above results demonstrated that the novel A2/O–La–Fe-MB system is a promising technique for highly-efficient TP removal from wastewater.

Performance enhancement and population structure of denitrifying phosphorus removal system over redox mediator at low temperature

The development of denitrifying polyphosphate accumulating organisms (DPAOs) presents a strategy to carbon competition between denitrifying bacteria and phosphorus removing bacteria. However, low temperature inhibits the rate of enzyme-catalyzed and substrate diffusion during denitrifying phosphorus removal (DPR). Therefore, the present study assessed the addition of NQS (100 μmol/L) for enhancing the removal of TP and TN in DPR reactors operated at alternating anaerobic and anoxic phases and different influent phosphate concentrations. The results showed that the removal efficiency of TP and TN in NQS-DPR system at 10 °C were 99.9% and 42.0%, respectively, which were 2.1 and 2.0 times higher than that of DPR system. Adding NQS significantly alleviated the increase of pH under anoxic condition and decreased the ORP value of the reactor, which in turn enhanced the PHAs accumulation process. The determination of functional genes (nirK, narG and phoD) showed that Dechloromonas, Lentimicrobium, and Terrimonas were the dominant functional bacteria in NQS-DPR system at 10 °C with the relative abundance of 3.09%, 2.99% and 2.28%, respectively. This study can provide valuable information for the effects of the addition of the redox mediator on denitrifying phosphorus removal technology.

Synergistic simultaneous endogenous partial denitrification/anammox (EPDA) and denitrifying dephosphatation for advanced nitrogen and phosphorus removal in a complete biofilm system

To achieve simultaneous biological nitrogen and phosphorus removal from municipal wastewater, the endogenous partial denitrification/anammox (EPDA) was combined with denitrifying dephosphatation in a complete biofilm reactor. Advanced nitrogen and phosphorus removal were achieved with effluent total nitrogen (TN) and PO43−-P concentrations of 7.77 ± 0.33 mg/L and 0.35 ± 0.10 mg/L, respectively. Anammox took a major role in the system, accounting for 76 ± 7% of nitrogen removal. 16S rRNA high-throughput sequencing results showed that the anammox bacteria co-existed with the denitrifying glycogen accumulating organisms (DGAOs) and the denitrifying phosphorus accumulating organisms (DPAOs). Anammox bacteria were mainly distributed in the inner layer, while DGAOs and DPAOs existed in the outer layer of EPDA biofilms. Furthermore, based on the EPDA biofilm system, a promising advanced nitrogen and phosphorus removal process was suggested to achieve lower requirements for energy and reagent consumption.

Insights into the denitrifying phosphorus removal decay processes by profiling of the response mechanism of denitrifying phosphate-accumulating organisms to starvation stress

Starvation conditions were inevitably encountered by biological wastewater treatment systems. Four anaerobic starvation periods (5, 10, 16 and 20 days) were conducted to investigate the response mechanism of denitrifying phosphate-accumulating organisms (DPAOs) in order to dissect denitrifying phosphorus removal (DPR) decay processes. The denitrifying phosphorus removal performance suffered with the decay rate of 0.162 ± 0.022 d−1 during 20-day starved duration. Metabolic activity decay was responsible 93.20 ± 0.11% for the damaged DPR performance, while biomass decay contributed to 6.79 ± 0.68%. The genus Dechloromonas affiliated to DPAOs exerted stronger survival adaptability to starvation with the abundance increasing from 1.98% to 3.15%, depended upon the endogenous consumption of intracellular polymers. In view of PHA-driven DPR mechanism of DPAOs, the metabolic activity was restricted by the depletion of available PHA. These results revealed the poorer stability but preponderant recovery of DPR system encountering with starvation.

Nitrogen removal pathways during simultaneous nitrification, denitrification, and phosphorus removal under low temperature and dissolved oxygen conditions

Nitrogen removal pathways of simultaneous nitrification, denitrification, and phosphorus removal (SNDPR) at low dissolved oxygen (0.3 mg/L) and temperature (10℃) were explored to understand nitrogen removal mechanisms. Biological nitrogen and phosphorus removal was sustained with total inorganic nitrogen removal, phosphorus removal, and simultaneous nitrification and denitrification (SND) efficiencies of 62.6%, 97.3%, and 31.2%, respectively. The SND was observed in the first 2 h of the aerobic phase and was attributed to denitrifying ordinary heterotrophic organisms using readily biodegradable chemical oxygen demand and denitrifying phosphorus accumulating organisms (DPAOs), which removed 15.1% and 12.2% of influent nitrogen, respectively. A phosphorus accumulating organism (PAO)-rich community was indicated by stoichiometric ratios and supported by 16S rRNA gene analysis, with Dechloromonas, Zoogloea, and Paracoccus as DPAOs, and Ca. Accumulibacter and Tetrasphaera as PAOs. Even though Ca. Competibacter (10.4%) was detected, limited denitrifying glycogen accumulating organism denitrification was observed.

Bioaugmentation for low C/N ratio wastewater treatment by combining endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) in the continuous A2/O - MBBR system

Endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) were combined in a novel A2/O - MBBR (Anaerobic Anoxic Oxic - Moving Bed Biofilm Reactor) system for low carbon/nitrogen (C/N) ratio wastewater treatment. The DPR performance was compared and the nutrient metabolism was elucidated based on the optimization of hydraulic retention time (HRT, 4–12 h) and nitrate recycling (R, 200%–600%). In the continuous-flow, the nitrate (NO3−) denitrification accompanied by nitrite (NO2−, via EPD) accumulation with the nitrate-to-nitrite transformation ratio (NTR) of 35.87%–43.31% in the anoxic zones. At HRT of 12 h with R of 500%, batch test initially revealed the DPR mechanism using both NO3− and NO2− as electron acceptor, where denitrifying phosphorus accumulation organisms (DPAOs) and denitrifying glycogen accumulation organisms (DGAOs) were the main contributors for EPD with incomplete denitrification (NO3− → NO2−). Furthermore, stoichiometry-based functional bacteria analysis displayed that higher bioactivity of DPAOs (NO2−→N2, 57.30%; NO3−→N2, 35.85%) over DGAOs (NO3−→N2, 6.85%) facilitated the anoxic NO3− reduction. Microbial community analysis suggested that Cluster I of Defluviicoccus-GAO group (∼4%) was responsible for stable NO2− accumulation performance via EPD, while increased Accumulibacter-PAO group (by ∼15%) contributed to the advanced nutrient removal. Based on the achievement of NO2− accumulation, the application feasibility of integrated EPD - DPR - Anammox for deep-level nutrient removal was discussed.

Simultaneous partial nitrification and denitrifying phosphorus removal (PNDPR) in a sequencing batch reactor process operated at low DO and high SRT for carbon and energy reduction

This study reports on a novel partial nitrification-denitrifying phosphorus sequencing batch reactor (SBR) system enriched with non-conventional denitrifying phosphorus accumulating organisms (DPAOs), treating carbon limited synthetic wastewater over 175 days. The non-conventional operating conditions, such as low DO (0.25 mg/L–0.35 mg/L) and relatively long solids retention time (SRT) of 15 d, favored the enrichment of a wide variety of denitrifying phosphorus accumulating organisms (DPAOs), such as Rhodocyclus, Dechloromonas, and Cytophaga. In contrast to the Accumulibacter, these microorganisms can sustain a very low DO environment and simultaneously perform denitrification and enhanced biological phosphorus removal (EBPR) using oxygen, nitrite, and nitrate as electron acceptors. The low DO also favored the washout of nitrite oxidizing bacteria (NOB), leading to simultaneous partial nitrification-denitrifying phosphorus removal (PNDPR). Partial nitrification at a high nitrite accumulation ratio (NAR ~ 80%) was also unfavourable for the growth of denitrifying glycogen accumulating organisms (DGAOs) in the PNDPR system. Simultaneous nitritation-denitritation (SND), nitrogen, and phosphorus removal efficiencies were maintained above 90%. Online cyclic tests and batch studies showed that nitrite shunt was achieved, and denitrification was predominantly controlled by DPAOs rather than ordinary heterotrophs (OHO). Mass balances showed that the COD/NOX−-N ratio for denitrification was 3.8. Compared to the conventional enhanced biological phosphorus removal (EBPR) process, the low DO-PNDPR process achieves reductions in carbon and aeration energy demands by approximately 47% relative to conventional biological nutrient removal (BNR) plants, and 49%, relative to conventional nitrification/denitrification systems, respectively.

Achieving and control of partial denitrification in anoxic-oxic process of real municipal wastewater treatment plant

Partial denitrification is an alternative process to provide stable nitrite for anammox. In this study, based on full-scale and lab-scale experiments, achieving and control of partial denitrification and the microbial mechanism were studied for 17 months in municipal wastewater treatment plant (MWWTP). Using glucose (GLC) as sole carbon source, partial denitrification was successfully achieved with nitrite accumulation percentage (NAP) higher than 90%; whereas, using sodium acetate (NaAc) as sole carbon source, nitrite accumulation was effectively controlled with economic and efficient carbon usage. Candidatus Competibacter and Thaurea were the dominant communities for partial denitrification. Denitrifying glycogen accumulating organisms (DGAOs), Thauera, denitrifying phosphorus accumulating organisms (DPAOs), GAOs, PAOs and denitrifiers coexisted in MWWTP, resulting in COD specific removal rate (CODSRR) of 883.10 ~ 1188.92mgN/gMLVSS/h during partial denitrification. Through adjustment of Anoxic-Oxic (A/O) operation to anoxic operation, the growth of GAOs and PAOs could be limited.

Ameliorating effect of nitrate on nitrite inhibition for denitrifying P-accumulating organisms

Lowered air supply and organic carbon need are the key factors to reduce wastewater treatment costs and thereby, avoid eutrophication. Denitrifying PO43−- removal (DPR) process using nitrate instead of oxygen for PO43− uptake was started up in the sequencing batch reactor (SBR) at a nitrate dosing rate of 20–25 mg N L−1 d−1. Operation with a real municipal wastewater supplied with CH3COONa, K2HPO4 and KNO3 succeeded in the cultivation of biomass containing denitrifying polyphosphate accumulating organisms (DPAOs). The durations of SBR process anaerobic/anoxic/oxic cycles were 1.5 h, 3.5 h and 1 h, respectively. SBR operation resulted in a maximum PO43−-P uptake of 17 mg PO43−-P g−1 MLSS. The highest TN and PO43− removal efficiencies were observed during the first half of reactor operation at 77 (±10) % and 71 (±5) %, respectively. An average COD removal rate of 172 (±98) mg g−1 MLSS and a high average removal efficiency of 89 (±4) % were achieved.Nitrite effect with/without nitrate as DPR electron acceptor was investigated in batch-scale to show possibilities to use high nitrite and nitrate contents simultaneously as electron acceptors for the anoxic phosphate uptake. Nitrate attenuation against nitrite toxicity can be economically justified in full-scale treatment applications in which wastewater has a high nitrogen content. Nitrate attenuated nitrite toxicity (caused by nitrite content at 5–100 mg NO2−-N L−1) when using supplemental additions of nitrate (at concentrations of 45–200 mg NO3−-N L−1) in batch tests. Illumina sequencing emphasized that during biomass adaption microbial community changed by lowered aerobic cycle length and by lowered nitrate dosing towards representation of key DPAO/PAO- organisms, such as Candidatus Accumulibacter, Xanthomonadaceae, Comomonadaceae, Saprospiraceae and Rhodocyclaceae. This study showed that DPAO biomass adaption to nitrate maintained an efficient COD, nitrogen and phosphorus removal and the biomass can be applied for treatment of wastewater containing high nitrite and nitrate content.

Optimization nutrient removal at different volume ratio of anoxic-to-aerobic zone in integrated fixed-film activated sludge (IFAS) system

This study evaluated the influence of different volume ratios of the anoxic-to-aerobic zone (Vano/Vaer) on the enhancement of nitrogen and phosphorus removal in an integrated fixed-film activated sludge (IFAS) system. As the Vano/Vaer increased from 1:2 to 2:1, the removal of organic carbon, nitrogen and phosphorus nutrients of the IFAS system was improved. At Vano/Vaer = 1:1, the removal effect of nitrogen and phosphorus nutrients was optimal, and the average removal rates of COD, NH4+-N, TN, and TP of the system reached 90 ± 3.2%, 98.2 ± 1.4%, 88.9 ± 2.2%, and 89.1 ± 2.7%, respectively. As the volume of the anoxic zone continued to increase, the denitrifying phosphate-accumulating capacity of the system was enhanced, and the highest ratio of specific anoxic and aerobic phosphorus uptake rate could reach 65.3%. Analysis of the molecular evaluation showed that, the proportion of nitrifying bacteria in the biofilm gradually increased as Vano increased. Moreover, denitrifying phosphate-accumulating organisms (DNPAOs), ammonia-oxidizing archaea (AOA), and anaerobic ammonium oxidizing (Anammox) bacteria were all enriched all showed enrichment in the biofilm of fiber carriers, which further strengthened the system's synergistic removal of nitrogen and phosphorus.

Nutrients removal by interactions between functional microorganisms in a continuous-flow two-sludge system (AAO-BCO): Effect of influent COD/N ratio

Denitrifying phosphorus removal (DPR) technology is one of the most effective approach to simultaneously realize nitrogen (N) and phosphorus (P) removal from low COD/N ratio wastewater. Identifying the interaction of denitrifying phosphate-accumulating organisms (DPAOs), denitrifying glycogen organisms (DGAOs) and denitrifying ordinary heterotrophic organisms (DOHOs) is critical for optimizing denitrification and anoxic P uptake efficiency in DPR processes. In this study, a novel DPR system of anaerobic anoxic oxic - biological contact oxidation (AAO-BCO) was employed to dispose actual sewage with various influent COD/N ratios (3.5–6.7). High efficiency of TIN (76.5%) and PO43−-P (94.4%) removal was observed when COD/N ratio was between 4.4 and 5.9. At the COD/N ratio of 5.7 ± 0.2, prominent DPR performance was verified by the superior DPR efficiency (88.7%) and anoxic phosphorus uptake capacity (PUADPAOs/ΔTIN = 1.84 mg/mg), which was further proved by the preponderance of DPAOs in C, N and P removal pathways. GAOs have a competitive advantage over PAOs for COD utilization at low COD/N ratio of 3.7 ± 0.2, which further limited the N removal efficiency. High proportion of N removal via DOHOs (21.2%) at the COD/N ratio of 6.5 ± 0.2 restrained the DPR performance, which should be attributed to the outcompete of DOHOs for NO3−. The nutrient removal mechanisms were explicated by stoichiometric calculation methodology to quantify the contribution of diverse functional microorganisms, contributing to improving the robustness of AAO-BCO system when facing the fluctuation of influent carbon source concentration.

Nitrite and nitrate inhibition thresholds for a glutamate-fed bio-P sludge

Enhanced biological phosphorus removal (EBPR) is an efficient and sustainable technology to remove phosphorus from wastewater. A widely known cause of EBPR deterioration in wastewater treatment plants (WWTPs) is the presence of nitrate/nitrite or oxygen in the anaerobic reactor. Moreover, most existing studies on the effect of either permanent aerobic conditions or inhibition of EBPR by nitrate or free nitrous acid (FNA) have been conducted with a “Candidatus Accumulibacter” or Tetrasphaera-enriched sludge, which are the two major reported groups of polyphosphate accumulating organisms (PAO) with key roles in full-scale EBPR WWTPs. This work reports the denitrification capabilities of a bio-P microbial community developed using glutamate as the sole source of carbon and nitrogen. This bio-P sludge exhibited a high denitrifying PAO (DPAO) activity, in fact, 56% of the phosphorus was uptaken under anoxic conditions. Furthermore, this mixed culture was able to use nitrite and nitrate as electron acceptor for P-uptake, being 1.8 μg HNO2–N·L−1 the maximum FNA concentration at which P-uptake can occur. Net P-removal was observed under permanent aerobic conditions. However, this microbial culture was more sensitive to FNA and permanent aerobic conditions compared to “Ca. Accumulibacter”-enriched sludge.

Advanced nitrogen and phosphorus removal by combining endogenous denitrification and denitrifying dephosphatation in constructed wetlands

To achieve high-efficiency nutrient removal in constructed wetlands (CWs), a novel simultaneous nitrogen and phosphorus removal (SNPR) process was developed by combining nitrification, endogenous denitrification, and denitrifying phosphorus removal. In SNPR process, denitrifying glycogen-accumulating organisms (DGAOs) and denitrifying polyphosphate-accumulating organisms (DPAOs) utilized NOx--N(NO3--N or NO2--N) as electron acceptor and poly-beta-hydroxy-alkanoates (PHAs) as carbon sources for endogenous denitrification and denitrifying phosphorus removal processes. Results from 217 days of operation showed that a high-level of nitrogen removal efficiency of 83.73% was achieved with influent COD/N of 4. The success was attributed to the fact that most of influent carbon sources could be transformed into PHAs before nitrification via enriching DGAOs and DPAOs in CW, which simultaneously improved nitrification and denitrification due to reducing oxygen and carbon sources consumption by aerobic heterotrophs. Phosphorus was mainly removed via denitrifying phosphorus removal, and PO43--P removal efficiency reached up to 87.84% with even common gravel used as substrate. Stoichiometry analysis revealed that DGAOs were the main organisms providing nitrite to DPAOs, suggesting that the effective PO43--P removal under high DGAO abundance condition might be attributed to the coordination of DGAOs and DPAOs in SNRP processes.

Simultaneous Domestication of Short-cut Nitrification Denitrifying Phosphorus Removal Granules

In this experiment, three replicated SBR reactors were operated using asynchronous acclimation of the phased method (A/O-A/O/A), simultaneous domestication of continuous aeration by A/O/A, and simultaneous domestication of intermittent aeration by A/O/A. Using artificial water distribution as the influent substrate, flocculent sludge was inoculated and granulated by hydraulic selection. The domestication and nitrogen and phosphorus removal characteristics of shortcut nitrification denitrifying phosphorus removal granules under different operation modes were assessed. The results show that simultaneous domestication of intermittent aeration by A/O/A has the most efficient under the combination of short aeration time (140 min) and low aeration strength[3.5 L·(h·L)-1]. The average removal rates of carbon, nitrogen, and phosphorus were 90.74%, 91.15%, and 95.66%, respectively, which could achieve synchronous removal during later stable operation. The particle size was 895 μm, and the particles were small but uniformly dense in microscope observations. The f value (MLVSS/MLSS) was kept stable at 0.8-0.85 and sludge had a high biomass due to the alternate aerobic/anoxic operation with intermittent aeration. This supported anoxic heterotrophic bacteria at the core of the particles, which was conducive to the stability of the granular sludge structure. Batch experiments showed that the specific ammonia-oxidation rate of the simultaneous domestication of intermittent aeration by A/O/A system was 3.38 mg·(g·h)-1, and denitrifying phosphate accumulating organisms (DPAOs) able to utilize nitrite as electron acceptor accounted for 65.46%. This was more conducive to the simultaneous domestication and enrichment of ammonia-oxidizing bacteria (AOB) and NO2--type DPAOs, ensuring a stable treatment effect.

Improvement of the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by nitrite stress

This study investigated a new way to improve the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by regularly changing the anaerobic/micro-aerobic/anoxic mode to the anaerobic/anoxic mode with 30 mg/L of nitrite dosing. The results indicated that the removal efficiency of total inorganic nitrogen and PO43−-P was improved from 75.44% and 85.14% to 98.89% and 98.17%, respectively. And the good performance of the SNDPR showed a long-time sustainability when the C/N ratio was 5. The results of microbial community illustrated that the abundance of the main nitrite-oxidizing bacteria (NOB), Nitrospira sp., dropped from 5.71% to 0.85% and the abundance of denitrifying polyphosphate-accumulating organisms (DPAOs), Pseudomonas sp. and Acinetobacter sp., increased by 5 times after nitrite stress. The high level of nitric oxide (NO) and free nitrite acid produced by addition of nitrite strongly suppressed the undesired organisms NOB and ordinary heterotrophic denitrifying organisms, and promoted the enrichment of DPAOs. The NO accumulated in the nitrite denitrification process could inhibit NOB and promote AOB. This study revealed that NO plays an important role in regulating the microbial community in the SNDPR system.

Denitrifying phosphorus removal and microbial community characteristics of two-sludge DEPHANOX system: Effects of COD/TP ratio

Two Lab-scale ANOXic DEPHosphation (DEPHANOX) bioreactors, in the continuous flow (DEPHANOX-CFR) and sequencing batch reactor (DEPHANOX-SBR) modes, were successfully operated and compared with varying influent C/P ratios. A decrease of influent chemical oxygen demand (COD)/total phosphorus (TP) feeding ratio from 250/4 to 250/14 generally did not significantly affect the nutrient removal performances. However, the overall microbial community structure and composition significantly shifted in the both two DEPHANOX bioreactors. Quantitative polymerase chain reaction results revealed that denitrifying phosphate-accumulating organisms (DPAOs) dominated and accounted for around 78.8 % of phosphorus accumulating organisms (PAOs). High-throughput sequencing results further suggested that the most three prominent phyla identified in the DEPHANOX system under different COD/TP ratio conditions included Proteobacteria (38.8–68.7 %), Bacteroidetes (10.3–12.3 %) and Chloroflexi (12.2−1.9 %). In addition, Dechloromonas and Pseudomonas spp. were identified as the dominant DPAOs. Furthermore, PICRUSt prediction analysis suggested that functional genes related to diverse metabolism pathways, including tricarboxylic acid cycle, electron transport, and nitrogen metabolism, were predicted to be actively involved with simultaneous phosphorus and nitrogen (SPN) removal. This study optimized the influent constituent characteristics and operating conditions of the DEPHANOX system, and provided novel insights into key metabolism functions of DPAOs in the DEPHANOX system.

Pairing denitrifying phosphorus accumulating organisms with anaerobic ammonium oxidizing bacteria for simultaneous N and P removal

Coupling of denitrifying polyphosphate accumulating organisms (DPAO) with anaerobic ammonium oxidizing (Anammox) bacteria in a single treatment scheme has so far been unsuccessful but could offer substantial energy savings, minimize sludge production, while achieving simultaneous carbon, nitrogen and phosphate removal. However, both organisms compete for nitrite and have vastly different growth rates and therefore the goal of this study was to uncouple their solid retention time (SRT) by growing them in different sludge fractions and to determine their biomass specific kinetic properties. Anammox bacteria were grown in a biofilm for longer SRTs and DPAO in flocs to allow shorter SRTs. Exposure of DPAO to anaerobic conditions was accomplished by recycling the flocs to a separate reactor by which simultaneous P, N, and C removal was accomplished. The diffusion limited biofilm lowered the biomass specific nitrite affinity for Anammox (KsAMX = 0.091 mM), which gave DPAO a competitive edge to consume nitrite (KsDPAO = 0.022 mM) in the suspended floc fraction. However, DPAO are more sensitive to nitrite (KiDPAO = 0.377 mM) than Anammox bacteria and (KiAMX > 1.786 mM), and therefore the DPAO would be better suited to grow in the protective biofilm, showing that both biomass growth types (flocs and granules) have advantages (and disadvantages) depending on the setting. This work is an important steppingstone to understanding resource competition amongst Anammox and DPAO and SRT management strategies to allow their pairing in combined reactor configurations.