Polyphosphate-accumulating Organisms Research Articles

C/N ratio effect on oily wastewater treatment using column type SBR: machine learning prediction and metagenomics study

The sequencing batch reactor has emerged as a promising technology in treating wastewater; however, its application in the treatment of generated water still needs to be explored. This research gap led to the investigation of various carbon-to-nitrogen (C/N) ratios in a column-type sequencing batch reactor (cSBR). The resulting data and model demonstrated that augmenting the SND process with an external carbon source is effective until the C/N ratio reaches 15, ultimately eliminating nitrogen in the produced water. Conversely, a reduced C/N ratio can limit the ability of polyphosphate-accumulating organisms to incorporate carbon into polyphosphate synthesis, thereby decreasing phosphorus removal efficiency within the cSBR. When the C/N ratio ranged from 6 to 8, and the mixed liquor suspended solids concentration was high, the average phosphate removal was approximately 55%, compared to only around 25% when the C/N ratio was less than 6.

Substrate preference of Tetrasphaera in dual PAOs synergistic EBPR system and its phosphorus removal performance

In this study, an alternated anaerobic/aerobic sequencing batch reactor (SBR) was employed to investigate the effect of the substrate conversion from casein hydrolysate (Cas aa) to a mixture of amino acids (glutamate, aspartate, glycine and proline) on the efficacy of nitrogen and phosphorus removal in an enhanced biological phosphorus removal (EBPR) system characterized by dual polyphosphate-accumulating organisms (PAOs), mainly focusing on the substrate preference of fermentative PAOs. The results indicated that a stable removal efficiency was found to be above 90 % as the substrate was converted to the mixture of amino acids. Meanwhile, the removal efficiency of ammonium eventually increased to over 95 % in stage III, despite an increase of ammonia nitrogen by hydrolysis with the rising proportion of mixed amino acids in influent. Moreover, shortening the anaerobic duration did not significantly affect the phosphorus removal performance, even enhancing anaerobic phosphorus release capacity. Nevertheless, the ammonia nitrogen removal was influenced. 16 s rRNA high-throughput sequencing results demonstrated that the detected fermentative PAO like Tetrasphaera in the EBPR system exhibited a clear preference for utilizing mixed amino acids and could cooperate with traditional PAO like Accumulibacter for phosphorus removal. In addition, the fermentation of amino acids mixture by Tetrasphaera was more pronounced and its abundance increased under the condition of a shorter anaerobic duration, leading to a relatively stable and high abundance of Accumulibacter in the presence of sufficient substrates from Tetrasphaera. However, considering the nitrogen removal performance and steady operation of the EBPR system, a longer anaerobic duration was more reasonable.

Efficient nitrogen and phosphorus removal performance and microbial community in a pilot-scale anaerobic/anoxic/oxic (AOA) system with long sludge retention time: Significant roles of endogenous carbon source

Stringent wastewater discharge standards require wastewater treatment plants (WWTPs) to focus on enhancing nitrogen (N) and phosphorus (P) removal efficiency. Increasing sludge concentration by regulation of sludge retention time (SRT) would enhance wastewater treatment loads. However, phosphorus-accumulating organisms (PAOs) would be outcompeted by glycogen-accumulating organisms (GAOs) under long SRT, leading to a collapse of P removal. In this work, pilot-scale anaerobic-oxic-anoxic (AOA) and anaerobic-anoxic-oxic (AAO) systems with long SRT (30 d) were parallelly established for actual urban wastewater treatment. The results indicated that sludge reflux ratio, temperature, and C/N ratio significantly impact N and P removal performance of AOA and AAO systems with long SRT, and removal efficiency of AOA system significantly exceeded that of AAO system. AOA system with long SRT achieved the optimal performance at sludge reflux ratio of 200%, temperature of 25 °C, and C/N ratio of 8, with COD, NH4+-N, TN, and PO43--P removal ratio of 92.80 ± 2.24%, 97.38 ± 0.89%, 88.97 ± 2.47%, and 94.33 ± 3.27%, respectively. In addition, compared to AAO system, AOA system could save 23.08% of the aeration volume. This work highlighted that AOA system with long SRT included multiple coupled nitrogen and phosphorus removal pathways, such as autotrophic/heterotrophic nitrification, anoxic/oxic denitrification, endogenous denitrification, and denitrifying phosphorus removal. Among these, the synergistic effect of endogenous denitrification and denitrifying phosphorus removal driven by internal carbon sources contributed to satisfactory nitrogen and phosphorus removal efficiency in AOA system with long SRT.

Macrogenomic analysis of tolerance and degradation mechanisms of polycyclic aromatic hydrocarbon in carbon and nitrogen metabolic pathways and associated bacterial communities during endogenous partial denitrification

The effective production of NO2−-N through endogenous partial denitrification (EPD) provides a promising perspective for the broader adoption and application of anaerobic ammonia oxidation. However, the accumulation of polycyclic aromatic hydrocarbons (PAHs) in the environment may worsen the operational challenges of the EPD system. This study evaluated the resilience of the EPD system to the toxic impacts of phenanthrene (PHE) and anthracene (ANT) through macrogenomic analysis. A control group was maintained under identical conditions for comparison. The results revealed that PHE and ANT had a relatively minimal impact on NO3−-N transformation and organic matter removal but significantly affected PO43−-P removal and NO2−-N accumulation in the EPD process. The PHE system achieved a higher NO2−-N accumulation, with a maximum NO3−-N to NO2−-N conversion ratio of 90.08%. In contrast, the ANT system exhibited higher efficiency in the PO43−-P removal, achieving a peak removal rate of 74.94%. Macrogenomic analysis revealed that PAHs significantly enriched both denitrifying glycogen-accumulating organisms (including Candidatus_Competibacter) and denitrifying polyphosphate-accumulating organisms (such as Thauera, Candidatus_Contendobacter, and Candidatus_Accumulibacter). This enrichment stabilized these organisms, facilitating NO2−-N accumulation and PO43−-P removal. Metabolic pathway analysis indicated that PHE promoted the enrichment of NO3−-N reductase and inhibited NO2−-N reductase activity. However, ANT stimulated oxidative phosphorylation and the phosphate cycle. Moreover, PAH metabolites enhanced the expression of key genes encoding succinate dehydrogenase and isocitrate dehydrogenase in the tricarboxylic acid cycle within the EPD process, leading to increase the synthesis and utilization of acetyl coenzyme-A. Notably, significant differences were observed between the effects of PHE and ANT on these metabolic processes. This study provides new methods for treating PAH-containing wastewater through the combination of EPD and anaerobic ammonia oxidation.

Candidatus Thiothrix phosphatis SCUT-1: A novel polyphosphate-accumulating organism abundant in the enhanced biological phosphorus removal system

A novel coccus Thiothrix-related polyphosphate-accumulating organism (PAO) was enriched in an acetate-fed enhanced biological phosphorus removal (EBPR) system. High EBPR performance was achieved for an extended period (>100 days). A high-quality draft genome (completeness 97.2 %, contamination 3.26 %) was retrieved, representing a novel Thiothrix species (with similarity<93.2 % to known Thiothrix species), and was denoted as 'Candidatus Thiothrix phosphatis SCUT-1'. Its acetate uptake rate (6.20 mmol C/g VSS/h) surpassed most Ca. Accumulibacter and known glycogen-accumulating organisms (GAOs), conferring their predominance in the acetate-fed system. Metatranscriptomic analysis suggested that Ca. Thiothrix phosphatis SCUT-1 employed both low- and high-affinity pathways for acetate activation, and both the conventional (PhaABC) pathway and the fatty acid β-oxidation pathway for PHA synthesis; additionally, a much more efficient FAD-dependent malate: quinone oxidoreductase (MQO) were encoded and employed than the traditional malate dehydrogenase (MDH) to oxidize malate to oxaloacetate in the TCA and glyoxylate cycle, collectively contributing to a higher acetate utilization and processing rate of this microorganism. Batch tests further demonstrated the versatile ability of this PAO in using VFA (acetate, propionate, and butyrate), lactate, amino acids (aspartate and glutamate), and glucose as carbon sources for EBPR, showing a partially overlapped but unique ecological niche of this microorganism comparing to Ca. Accumulibacter and known GAOs. A metabolic model was built for Ca. Thiothrix phosphatis SCUT-1 using the above-mentioned carbon sources for EBPR. Overall, this study represents the first comprehensive characterization of the physiology and metabolic characteristics of representative coccus Thiothrix-related PAOs, which are expected to provide new insights into PAO microbiology in EBPR systems.

Transcriptional profiling elucidates biofilm functionality in the dynamic environment of an enhanced biological phosphorus removal reactor

ABSTRACT Recently, biofilms, complex and dynamic structures of microorganisms, have been applied to enhanced biological phosphorus removal (EBPR), a wastewater treatment configuration dependent on cyclic shifts between anaerobic and aerobic conditions. In this study, comparative metagenomics and metatranscriptomics were performed on biofilms collected from seven sites of a moving-bed-biofilm-reactor-based EBPR process. The aim was to examine the functional ecology of phosphorus-accumulating biofilms throughout a single EBPR cycle. Taxonomic profiling revealed high microbial diversity, stable throughout the EBPR cycle. The dominant phosphorus-accumulating organisms (PAOs) were identified as Candidatus accumulibacter, Candidatus phosphoribacter, and Candidatus lutibacillus. However, these did not show the highest transcriptional activities. Propionivibrio, a potential glycogen-accumulating organism, was the most transcriptionally active. Comparative analysis of biofilms from different EBPR stages showed a progressive change in metatranscriptome composition, correlating with nutrient removal. Analysis of differentially expressed genes in abundant PAOs revealed key genes associated with the uptake of phosphorus, degradation of glycogen, biosynthesis of polyhydroxyalkanoates, and acetate production. In conclusion, this study reveals that biofilms possess the capability to adapt to environmental fluctuations primarily through alterations in microbial gene expression activity and subsequent metabolic modulation, and dominant taxa may not necessarily exhibit the highest transcriptional activity in complex microbial communities.

Thauera enriched granular activated carbon-assisted aerobic granular sludge performs enhanced biological nitrogen and phosphate removal via propionate utilization

Propionate constituting up to 25 % of volatile fatty acids in municipal wastewater, presents metabolic challenges for microorganisms. This study investigated the combined use of propionate along with granular activated carbon (GAC) for aerobic granular sludge (AGS) development and biological nutrient removal (BNR), particularly bio-P removal in granular reactors. GAC addition decreased granulation time and facilitated stable BNR pathways with propionate. AGS demonstrated robust settling properties (≥1 mm, 6 g/L MLSS, 40 mL/g SVI) with increased alginate-like exopolysaccharide (ALE) content up to 400 mgALE/gSS, indicating stable granule formation. Effective ammonium removal via simultaneous nitrification and denitrification was achieved. By day 90, phosphate removals reached 89 %, due to enrichment of polyphosphate accumulating organisms (PAOs) in GAC-containing AGS. The sludge fraction (>0.5 mm) comprised of granules and GAC-biofilms showed efficient phosphate removal through enhanced biological phosphate removal (EBPR). Enrichment of denitrifying PAO, Thauera sp., contributed to enhanced granulation, granular stability and nutrient removals.

Integrated process of Tetrasphaera-dominated in-situ waste activated sludge fermentation, biological phosphorus removal and endogenous denitrification in single reactor for treatment of wastewater with limited carbon sources

An integrated process of sludge in-situ fermentation, biological phosphorus removal and endogenous denitrification (ISFPR-ED) was developed to treat low ratio of chemical oxygen demand to nitrogen (COD/N) wastewater and waste activated sludge (WAS) in a single reactor. Nutrient removal and WAS reduction were achieved due to Tetrasphaera-dominated sludge fermentation provided organic carbon in extending the anaerobic duration. The WAS reduction efficiency, effluent orthophosphate (PO43−-P) and total inorganic nitrogen reached 28.1 %, less than 0.4 and 7.2 mg/L, respectively. While organic carbon was reduced by 67 %. Tetrasphaera, conventional polyphosphate accumulating organisms (PAOs) stored glycogen, amino acids, and PHA for nutrient removal. Excess energy from fermentation enhanced anaerobic PO43−-P uptake by Tetrasphaera. Tetrasphaera was the dominant PO43−-P removal and fermentation bacteria, working synergistically with conventional PAOs and fermenting microorganisms. This integrated process improves nutrient removal efficiency and reduces operating costs for carbon addition and WAS disposal in wastewater treatment.

Integrating enhanced biological phosphorus removal in adsorption-stage to treat real domestic sewage

Wastewater treatment innovation toward resource recovery facilities raises concerns about the adsorption and bio-degradation (A-B) process. This study integrated enhanced biological phosphorus removal (EBPR) into the A-stage for real domestic sewage treatment using the short sludge retention time (S-SRT) approach. The S-SRT approach resulted in outstanding phosphorus (over 90 %) and COD removal (approximately 88 %), increased sludge yield and organic matter content, and a 1.68-fold increase in energy recovery efficiency by sludge anaerobic digestion. The inhibition of nitrification relieved competition for carbon sources between denitrification and phosphorus removal, allowing for the enrichment of phosphorus-accumulating organisms (PAOs) such as Tetrasphaera and Halomonas, leading to enhanced phosphorus removal activities. Biological adsorption also plays a significant role in achieving steady phosphorus removal performance. This study demonstrates the potential of the S-SRT approach as an effective strategy for simultaneous carbon and phosphorus capture in the A-stage, contributing to energy and nutrient recovery from sewage.

Distinct microdiversity of phosphate accumulating organisms (PAOs) between side-stream and conventional enhanced biological phosphorus removal (EBPR) systems with performance implications

Polyphosphate Accumulating Organisms (PAOs) microdiversity is a key factor to elucidate the mechanisms involved in the side-stream enhanced biological phosphorus removal (S2EBPR) systems, which has been shown to improve the process stability over conventional EBPR. However, fast, effective and cost-efficient methods to resolve PAO microdiversity in real-world activate sludge samples is still in absence. In this study, we applied oligotyping analysis following the regular 16S rRNA gene amplicon sequencing standard operation pipeline (SOP) to resolve subgenus-level PAO oligotypes, which cannot be achieved using traditional 16S rRNA sequencing SOP. The identified oligotype profiles of PAO-containing genera Ca. Accumulibacter, Tetrasphaera and Comamonas showed distinguished community-level differences across 12 water resource recovery facilities (WRRFs), which would not be revealed at the genus level. The WRRF-level differences were observed larger than the temporal differences in the same WRRF, indicating intrinsic sub-genus level microdiversity fingerprint between EBPR/S2EBPR systems. The identified oligotypes can be associated with known PAO clades phylogenetically, suggesting that oligotyping can suffice as a fast and cost-efficient approach for PAO microdiversity profiling. In addition, network analysis can be used to identify coexistence patterns between oligotypes with respect to EBPR/S2EBPR configurations and performance, enabling more detailed analysis between EBPR system performance and PAOs microdiversity. Correlation analyses between oligotype profiles and key EBPR performance parameters revealed potential different biological functional traits among these PAO species with P-removal performance implications.

Orthophosphate uptake onto woodchips in denitrifying bioreactors is enhanced by anoxic-(anaerobic-)oxic cycling

Woodchips are widely used as a low-cost and renewable organic carbon source for denitrifying biofilms in passive nutrient removal systems. One limitation of wood-based biofiltration systems is their relatively poor removal of phosphorus (P) from subsurface drainage and stormwaters, necessitating the use of additional filter media when co-treatment of nitrogen (N) and P is required. Here, we show that anoxic-oxic cycling of woodchip media, which enhances nitrate (NO3−) removal by increasing the mobilization of organic carbon from wood, also improves orthophosphate (Pi) uptake onto woodchips. Orthophosphate removal rates in flow-through woodchip columns ranged from 0 to 34.9 μg PO43− L−1 h−1 under continuously-saturated (anoxic) conditions, and increased to 17.5 to 71.9 μg PO43− L−1 h−1 in columns undergoing drying-rewetting (oxic-anoxic) cycles. The highest Pi removal efficiencies were observed in the first 20 h after reactors were re-flooded, and were concurrent with maxima in polyphosphate kinase (ppk) gene expression by the polyphosphate accumulating organisms (PAOs) Accumulibacter spp. and Pseudomonas spp. Batch experiments confirmed that anoxic-anaerobic-oxic pre-incubation conditions led to orthophosphate uptake onto woodchips as high as 74.9 ± 0.8 mg PO43−/kg woodchip, and batch tests with autoclaved woodchips demonstrated that Pi uptake was due to biological processes and not adsorption. NO3− removal in batch tests was also greatest under oxic incubation conditions, attributed to greater carbon availability in hypoxic to anoxic zones in woodchip biofilms. While further research is needed to elucidate the mechanisms controlling enhanced Pi uptake by woodchip biofilms under anoxic-(anaerobic-)oxic cycling, these results suggest a role for enhanced Pi uptake by PAOs in a nature-based system for treatment of nonpoint source nutrients.

Outdoor tubular photobioreactor microalgae-microorganisms biofilm treatment of municipal wastewater: Enhanced heterotrophic assimilation and synergistic aerobic denitrogenation

This research evaluated a microalgae consortium (MC) in a pilot-scale tubular photobioreactor for municipal wastewater (MWW) treatment, compared with an aeration column photobioreactor. Transitioning from suspended MC to a microalgae-microbial biofilm (MMBF) maintained treatment performance despite increasing influent from 50 L to 150 L in a 260 L system. Carbon and nitrogen removal were effective, but phosphorus removal varied due to biofilm shading and the absence of phosphorus-accumulating organisms. High influent flow caused MMBF detachment due to shear stress. Stabilizing and re-establishing the MMBF showed that a stable phycosphere influenced microbial diversity and interactions, potentially destabilizing the MMBF. Heterotrophic nitrification-aerobic denitrification bacteria were crucial for MC equilibrium. Elevated gene expression related to nitrogen fixation, organic nitrogen metabolism, and nitrate reduction confirmed strong microalgal symbiosis, highlighting MMBF’s treatment potential. This study supports the practical application of microalgae in wastewater treatment.

Nitrate and nitrite utilization during denitrifying phosphorus removal: Electron acceptor preference and feasible process combinations

Denitrifying phosphorus removal (DPR), which is dominated by denitrifying polyphosphate-accumulating organisms (DPAOs), is a promising process for nitrogen and phosphorus removal. Denitrifying glycogen-accumulating organisms (DGAOs) and DPAOs typically coexist in the DPR sludge, complicating the study of DPAOs’ denitrification capacity. In this study, two reactors were fed with nitrate and nitrite during the anoxic phase to cultivate nitrate-DPR and nitrite-DPR sludge. Both reactors yielded high and low DGAO abundance sludges, enabling the evaluation of the denitrification capacity of DPAOs. For the nitrate-DPR sludge, the nitrite reduction rate was 1.63 times higher than the nitrate reduction rate when DPAOs were the primary denitrifiers. For the nitrite-DPR sludge, the reduction rate of nitrite was more than three times that of nitrate, irrespective of DGAO abundance. These findings indicated that DPAOs preferred nitrite to nitrate and were well suited to reduce nitrite rather than reduce nitrate to supply nitrite.

Nutrient removal and microbial community succession in moving bed biofilm reactor: Effects of influent carbon to nitrogen ratio fluctuation

This study investigated the nutrient removal and microbial community succession in moving bed biofilm reactor under stable and three levels of influent carbon/nitrogen (C/N) ratio fluctuation (± 10%, ± 20%, and ± 30%). Under the conditions of influent C/N ratio fluctuation, the removal efficiency of COD and PO43−-P decreased 4.7–6.4% and 3.7–12.9%, respectively, while the nitrogen removal was almost unaffected. A sharp decrease in the content of culturable functional bacteria related to nitrogen and phosphorus removal including nitrite-oxidizing bacteria (NOB), aerobic denitrifying bacteria (DNB), and polyphosphate-accumulating organisms (PAOs) from the carrier biofilm was observed. Sequencing analysis revealed that the abundance of Candidatus Competibacter increased 10.3–25.9% and became the dominant genus responsible for denitrification, potentially indicating that nitrate was removed via endogenous denitrification under the influent C/N ratio fluctuation. The above results will provide basic data for the nutrient removal in decentralized wastewater treatment under highly variable influent conditions.

Inhibition of phosphorus removal performance in activated sludge by Fe(III) exposure: transitions in dominant metabolic pathways.

Simultaneous chemical phosphorus removal process using iron salts (Fe(III)) has been widely utilized in wastewater treatment to meet increasingly stringent discharge standards. However, the inhibitory effect of Fe(III) on the biological phosphorus removal system remains a topic of debate, with its precise mechanism yet to be fully understood. Batch and long-term exposure experiments were conducted in six sequencing batch reactors (SBRs) operating for 155 days. Synthetic wastewater containing various Fe/P ratios (i.e., Fe/P = 1, 1.2, 1.5, 1.8, and 2) was slowly poured into the SBRs during the experimental period to assess the effects of acute and chronic Fe(III) exposure on polyphosphate-accumulating organism (PAO) growth and phosphorus metabolism. Experimental results revealed that prolonged Fe(III) exposure induced a transition in the dominant phosphorus removal mechanism within activated sludge, resulting in a diminished availability of phosphorus for bio-metabolism. In Fe(III)-treated groups, intracellular phosphorus storage ranged from 3.11 to 7.67 mg/g VSS, representing only 26.01 to 64.13% of the control. Although the abundance of widely reported PAOs (Candidatus Accumulibacter) was 30.15% in the experimental group, phosphorus release and uptake were strongly inhibited by high dosage of Fe(III). Furthermore, the abundance of functional genes associated with key enzymes in the glycogen metabolism pathway increased while those related to the polyphosphate metabolism pathway decreased under chronic Fe(III) stress. These findings collectively suggest that the energy generated from polyhydroxyalkanoates oxidation in PAOs primarily facilitated glycogen metabolism rather than promoting phosphorus uptake. Consequently, the dominant metabolic pathway of communities shifted from polyphosphate-accumulating metabolism to glycogen-accumulating metabolism as the major contributor to the decreased biological phosphorus removal performance.

The phototrophic metabolic behaviour of Candidatus accumulibacter

The phototrophic capability of Candidatus Accumulibacter (Accumulibacter), a common polyphosphate accumulating organism (PAO) in enhanced biological phosphorus removal (EBPR) systems, was investigated in this study. Accumulibacter is phylogenetically related to the purple bacteria Rhodocyclus from the family Rhodocyclaceae, which belongs to the class Betaproteobacteria. Rhodocyclus typically exhibits both chemoheterotrophic and phototrophic growth, however, limited studies have evaluated the phototrophic potential of Accumulibacter. To address this gap, short and extended light cycle tests were conducted using a highly enriched Accumulibacter culture (95%) to evaluate its responses to illumination. Results showed that, after an initial period of adaptation to light conditions (approximately 4–5 h), Accumulibacter exhibited complete phosphorus (P) uptake by utilising polyhydroxyalkanoates (PHA), and additionally by consuming glycogen, which contrasted with its typical aerobic metabolism. Mass, energy, and redox balance analyses demonstrated that Accumulibacter needed to employ phototrophic metabolism to meet its energy requirements. Calculations revealed that the light reactions contributed to the generation of, at least more than 67% of the ATP necessary for P uptake and growth. Extended light tests, spanning 21 days with dark/light cycles, suggested that Accumulibacter generated ATP through light during initial operation, however, it likely reverted to conventional anaerobic/aerobic metabolism under dark/light conditions due to microalgal growth in the mixed culture, contributing to oxygen production. In contrast, extended light tests with an enriched Tetrasphaera culture, lacking phototrophic genes in its genome, clearly demonstrated that phototrophic P uptake did not occur. These findings highlight the adaptive metabolic capabilities of Accumulibacter, enabling it to utilise phototrophic pathways for energy generation during oxygen deprivation, which holds the potential to advance phototrophic-EBPR technology development.

Synergistic Integration of Anammox and Endogenous Denitrification Processes for the Simultaneous Carbon, Nitrogen, and Phosphorus Removal.

The feasibility of a synergistic endogenous partial denitrification-phosphorus removal coupled anammox (SEPD-PR/A) system was investigated in a modified anaerobic baffled reactor (mABR) for synchronous carbon, nitrogen, and phosphorus removal. The mABR comprising four identical compartments (i.e., C1-C4) was inoculated with precultured denitrifying glycogen-accumulating organisms (DGAOs), denitrifying polyphosphate-accumulating organisms, and anammox bacteria. After 136 days of operation, the chemical oxygen demand (COD), total nitrogen, and phosphorus removal efficiencies reached 88.6 ± 1.0, 97.2 ± 1.5, and 89.1 ± 4.2%, respectively. Network-based analysis revealed that the biofilmed community demonstrated stable nutrient removal performance under oligotrophic conditions in C4. The metagenome-assembled genomes (MAGs) such as MAG106, MAG127, MAG52, and MAG37 annotated as denitrifying phosphorus-accumulating organisms (DPAOs) and MAG146 as a DGAO were dominated in C1 and C2 and contributed to 89.2% of COD consumption. MAG54 and MAG16 annotated as Candidatus_Brocadia (total relative abundance of 16.5% in C3 and 4.3% in C4) were responsible for 74.4% of the total nitrogen removal through the anammox-mediated pathway. Functional gene analysis based on metagenomic sequencing confirmed that different compartments of the mABR were capable of performing distinct functions with specific advantageous microbial groups, facilitating targeted nutrient removal. Additionally, under oligotrophic conditions, the activity of the anammox bacteria-related genes of hzs was higher compared to that of hdh. Thus, an innovative method for the treatment of low-strength municipal and nitrate-containing wastewaters without aeration was presented, mediated by an anammox process with less land area and excellent quality effluent.

Enhanced Simultaneous Nitrogen and Phosphorus Removal in a Continuous-Flow Granular Sludge System under Gradient-Controlled Hydraulic Loading

The feasibility of the aerobic granulation of activated sludge was investigated in a continuous-flow anaerobic–anoxic–oxic system under gradient-controlled hydraulic loading on the surface of a cyclone separator. Concentrated domestic sewage was used. After 80 days of operation, 80% of activated sludge in the system was in the form of granular sludge with an average particle size of 373 μm. High removal efficiency was achieved for chemical oxygen demand (94.40%), NH4+-N (99.93%), total nitrogen (89.44%), and total phosphorus (96.92%). A batch study revealed that Pseudomonas (1.34%) and Dechloromonas (1.05%) as the main denitrifying phosphorus-accumulating organisms could efficiently remove phosphorus using nitrate as an electron acceptor, which improved the utilization efficiency of carbon sources and achieved simultaneous denitrification and phosphorus removal. Overall, the study demonstrates the feasibility of enhanced denitrification and phosphorus removal in a continuous-flow granular sludge system. The sludge system enables simultaneous nitrogen and phosphorus removal under low carbon-to-nitrogen ratios.

Microbial community structure of an anaerobic side-stream coupled anoxic-aerobic membrane bioreactor (AOMBR-ASSR) for an in-situ sludge reduction process.

With the anoxic-aerobic membrane bioreactor (AO-MBR, CP) as a reference, high-throughput sequencing technology was used to reveal the characteristics of the microbial community structure in the anaerobic side-stream anoxic-aerobic membrane bioreactor sludge reduction process (AOMBR-ASSR, SRP). After the stable operation of two processes for 120days, the average removal efficiencies of TN and TP in the effluent of SRP were increased by 5.6% and 29.8%, respectively. The observed sludge yields (Yobs) of the two processes were 0.14 and 0.17gMLSS/(gCOD), respectively, and the sludge reduction rate of the SRP was 19.5%. Compared to the CP, the microbial richness and diversity index of SRP increased significantly. Chloroflexi, which is responsible for the degradation of organic substances under an anaerobic condition, seemed to be reduced in the SRP. Meanwhile, other phyla that involved in the nitrogen cycle, such as Nitrospirae and Planctomycetes, were found to be more abundant in the SRP than in the CP. A total of 21 identified classes were observed, and primarily hydrolyzed fermented bacteria (Sphingobacteriia, Betaproteobacteria, Actinobacteria and Deltaproteobacteria) and slow-growing microorganisms (Bacilli) were accumulated in the SRP. At the genus level, the inserted anaerobic side-stream reactor favored the hydrolyzed bacteria (Saprospiraceae, Rhodobacter and Candidatus_Competibacter), fermented bacteria (Lactococcus and Trichococcus), and slow-growing microorganisms (Dechloromonas and Haliangium), which play a crucial role in the sludge reduction. Furthermore, the enrichment of bacterial species related to nitrogen (Nitrospir and Azospira) provided the potential for nitrogen removal, while the anaerobic environment of the side-stream reactor promoted the enrichment of phosphorus-accumulating organisms.

Distinguished denitrifying phosphorus removal in the high-rate anoxic/microaerobic system for sewage treatment

This study re-evaluated the role of anoxic and anaerobic zones during the enhanced biological phosphorus (P) removal process by investigating the potential effect of introducing an anoxic zone into a high-rate microaerobic activated sludge (MAS) system (1.60–1.70 kg chemical oxygen demand (COD) m−3 d−1), i.e., a high-rate anoxic/microaerobic (A/M) system for sewage treatment. In the absence of a pre-anaerobic zone, introducing an anoxic zone considerably reduced effluent NOx−-N concentrations (7.2 vs. 1.5 mg L−1) and remarkably enhanced total nitrogen (75% vs. 89%) and total P (18% vs. 60%) removal and sludge P content (1.48% vs. 1.77% (dry weight)) due to further anoxic denitrifying P removal in the anoxic zone (besides simultaneous nitrification and denitrification in the microaerobic zone). High-throughput pyrosequencing demonstrated the niche differentiation of different polyphosphate accumulating organism (PAO) clades (including denitrifying PAO [DPAO] and non-DPAO) in both systems. Introducing an anoxic zone considerably reduced the total PAO abundance in sludge samples by 42% and modified the PAO community structure, including 17–19 detected genera. The change was solely confined to non-DPAOs, as no obvious change in total abundance or community structure of DPAOs including 7 detected genera was observed. Additionally, introducing an anoxic zone increased the abundance of ammonia-oxidizing bacteria by 39%. The high-rate A/M process provided less aeration, higher treatment capacity, a lower COD requirement, and a 75% decrease in the production of waste sludge than the conventional biological nutrient removal process.