Glycogen-accumulating Organisms Research Articles

Enhanced Low-Energy Chemical Oxygen Demand (COD) Removal in Aeration-Free Conditions through Pulse-Rotating Bio-Contactors Enriched with Glycogen-Accumulating Organisms

This study presents an innovative pulse-rotating biological contactor (P-RBC) designed to enrich glycogen-accumulating organisms (GAOs), thereby facilitating low-energy chemical oxygen demand (COD) removal. It then investigates the impact of rotational speed and hydraulic retention time (HRT) on GAO enrichment and COD removal efficiency. Optimized conditions at lower speeds and longer HRTs significantly enhance GAO proliferation and Polyhydroxyalkanoate (PHA) synthesis, the key to COD removal. Noteworthy findings include a maximum GAO abundance of 21.34% at a half round per hour (rph) rotating speed, which correlates with a 90.2% COD removal rate and an HRT of 6 h, yielding a 21.23% GAO abundance and 89.8% COD removal. This study also explores various carbon sources for PHA synthesis, with sodium acetate proving the most effective. Compared to other wastewater treatment methods, P-RBC demonstrates minimal energy consumption (0.09 kWh per ton of wastewater), highlighting its potential as a sustainable and effective approach for wastewater treatment.

Microbiome structure and function in parallel full-scale aerobic granular sludge and activated sludge processes

Aerobic granular sludge (AGS) and conventional activated sludge (CAS) are two different biological wastewater treatment processes. AGS consists of self-immobilised microorganisms that are transformed into spherical biofilms, whereas CAS has floccular sludge of lower density. In this study, we investigated the treatment performance and microbiome dynamics of two full-scale AGS reactors and a parallel CAS system at a municipal WWTP in Sweden. Both systems produced low effluent concentrations, with some fluctuations in phosphate and nitrate mainly due to variations in organic substrate availability. The microbial diversity was slightly higher in the AGS, with different dynamics in the microbiome over time. Seasonal periodicity was observed in both sludge types, with a larger shift in the CAS microbiome compared to the AGS. Groups important for reactor function, such as ammonia-oxidising bacteria (AOB), nitrite-oxidising bacteria (NOB), polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs), followed similar trends in both systems, with higher relative abundances of PAOs and GAOs in the AGS. However, microbial composition and dynamics differed between the two systems at the genus level. For instance, among PAOs, Tetrasphaera was more prevalent in the AGS, while Dechloromonas was more common in the CAS. Among NOB, Ca. Nitrotoga had a higher relative abundance in the AGS, while Nitrospira was the main nitrifier in the CAS. Furthermore, network analysis revealed the clustering of the various genera within the guilds to modules with different temporal patterns, suggesting functional redundancy in both AGS and CAS.Key points• Microbial community succession in parallel full-scale aerobic granular sludge (AGS) and conventional activated sludge (CAS) processes.• Higher periodicity in microbial community structure in CAS compared to in AGS.• Similar functional groups between AGS and CAS but different composition and dynamics at genus level.Graphical abstract

A Continuous Plug-Flow Anaerobic-Multistage Anoxic/Aerobic Process Treating Low-C/N Domestic Sewage: Nutrient Removal, Greenhouse Gas Emissions, and Microbial Community Analysis

The anaerobic-multistage anoxic/aerobic (A-MAO) process has shown good potential for advanced nitrogen removal in recent years, but its greenhouse gas emissions still need to be fully explored. The effects of the influent distribution and external carbon source sodium acetate on nutrient removal, greenhouse gas emissions, and the microbial community structure in a continuous plug-flow A-MAO reactor fed with real low C/N ratio domestic sewage were investigated. The results showed that altering the allocation of carbon source resulted in average chemical oxygen demand (COD) and total nitrogen (TN) concentration in effluent reduced to 26.10 ± 4.86 and 6.65 ± 1.73 mg/L, respectively. Both operations reduced the emission rate of greenhouse gas. While the addition of external car-bon sources leaded to lower N2O emission rates and higher CO2 and CH4 emission rates. The addition of sodium acetate facilitated nitrification and denitrification processes, thereby leading to a reduction in N2O production. Meanwhile, it spurred the growth of methanogenic bacteria and heterotrophic microorganisms, thus boosting the production of CO2 and CH4. Influent distribution promoted the increase of Bacteroidota, Chloroflexi and Acidobacteriota of the reactor. The enrichment of typical hydrolytic bacteria and glycogen accumulating organisms (GAOs) increased the utilization efficiency of carbon sources in the system after the addition of sodium acetate. The significant increase of typical denitrifying bacteria (DNBs) Azospira reduced the N2O emission during heterotrophic denitrification process, which was considered to be an important functional genus for increasing nitrogen loss in this system. The rational utilization of carbon source makes the difference in metabolism function. The study provides a valuable strategy for comprehensively evaluating the pollutant removal and greenhouse gas emission reduction from the A-MAO process.

Turning alkalinity deficiency-induced metabolic disorder into an opportunity for promoting granulation and enhancing heat generation

Shortage of alkalinity is a common problem in treating low carbon/nitrogen (C/N) wastewater and industrial influents. It negatively affects system stability and efficiency, but its positive effects remain unclear. To address this issue, insufficient (R1: 400 mg CaCO3/L) and sufficient alkalinity (R2: 750 mg CaCO3/L) levels were established in our study, where R1 served as the experimental group and R2 as the control group. In the experimental group, alkalinity deficiency deteriorated the nitrification and phosphorus removal process by reducing the abundance of functional microbes (Nitrosomonas, genus ammonia-oxidizing bacteria, AOB, and Dechloromonas, genus polyphosphate-accumulating organisms, PAOs) and genes (hao). At the same time, the alkalinity deficiency enriched slow-growing functional microbes (Candidatus_Competibacter, genus glycogen-accumulating organisms, GAOs) and key genes (nirK, norB, nosZ, acnB, fumA, fumB) increased the content of extracellular polymeric substances (EPS) and the ratio of protein to polysaccharide (PN/PS). By day 113, over 87 % of the particles were larger than 200 μm, and complete granulation was achieved. Microbial energy metabolism further indicated that alkalinity deficiency diverted some energy to non-growth metabolism, thereby enhancing microbial heat generation. Our results highlight an innovative approach, using the setting of insufficient alkalinity conditions to improve heat recovery and address the challenges of WWTPs. These research findings are in favor of wastewater treatment plants seeking to meet multiple sustainable objectives, including improving water quality, saving footprint, and maximizing energy recovery.

Cultivation of phosphate-accumulating biofilm: Study of the effects of acyl-homoserine lactones (AHLs) and cyclic dimeric guanosine monophosphate (c-di-GMP) on the formation of biofilm and the enhancement of phosphate metabolism capacity

This study investigated the mechanisms of microbial growth and metabolism during biofilm cultivation in the biofilm sequencing batch reactor (BSBR) process for phosphate (P) enrichment. The results showed that the sludge discharge was key to biofilm growth, as it terminated the competition for carbon (C) source between the nascent biofilm and the activated sludge. For the tested reactor, after the sludge discharge on 18 d, P metabolism and C source utilization improved significantly, and the biofilm grew rapidly. The P concentration of the recovery liquid reached up to 157.08 mg/L, which was sufficient for further P recovery via mineralization. Meta-omics methods were used to analyze metabolic pathways and functional genes in microbial growth during biofilm cultivation. It appeared that the sludge discharge activated the key genes of P metabolism and inhibited the key genes of C metabolism, which strengthened the polyphosphate-accumulating metabolism (PAM) as a result. The sludge discharge not only changed the types of polyphosphate-accumulating organisms (PAOs) but also promoted the growth of dominant PAOs. Before the sludge discharge, the necessary metabolic abilities that were spread among different microorganisms gradually concentrated into a small number of PAOs, and after the sludge discharge, they further concentrated into Candidatus_Contendobacter (P3) and Candidatus_Accumulibacter (P17). The messenger molecule C-di-GMP, produced mostly by P3 and P17, facilitated P enrichment by regulating cellular P and C metabolism. The glycogen-accumulating organism (GAO) Candidatus_Competibacter secreted N-Acyl homoserine lactones (AHLs), which stimulated the secretion of protein in extracellular polymeric substances (EPS), thus promoting the adhesion of microorganisms to biofilm and improving P metabolism via EPS-based P adsorption. Under the combined action of the dominant GAOs and PAOs, AHLs and C-di-GMP mediated QS to promote biofilm development and P enrichment. The research provides theoretical support for the cultivation of biofilm and its wider application.

A comprehensive floc model for simulating simultaneous nitrification, denitrification, and phosphorus removal

A comprehensive floc model for simultaneous nitrification, denitrification, and phosphorus removal (SNDPR) was designed, incorporating polyphosphate-accumulating organisms (PAOs), glycogen-accumulating organisms (GAOs), intrinsic half-saturation coefficients, and explicit external mass transfer terms. The calibrated model was able to effectively describe experimental data over a range of operating conditions. The estimated intrinsic half-saturation coefficients of oxygen values for ammonia-oxidizing bacteria, nitrite-oxidizing bacteria, ordinary heterotrophic organisms (OHOs), PAOs, and GAOs were set at 0.08, 0.18, 0.03, 0.07, and 0.1 mg/L, respectively. Simulation suggested that low dissolved oxygen (DO) environments favor K-strategist nitrifying bacteria and PAOs. In SNDPR, virtually all influent and fermentation-generated volatile fatty acids were assimilated as polyhydroxyalkanoates by PAOs in the anaerobic phase. In the aerobic phase, PAOs absorbed 997 % and 171 % of the benchmark influent total phosphorus mass loading through aerobic growth and denitrification via nitrite. These high percentages were because they were calculated relative to the influent total phosphorus, rather than total phosphorus at the end of the anaerobic period. When considering simultaneous nitrification and denitrification, about 23.1 % of influent total Kjeldahl nitrogen was eliminated through denitrification by PAOs and OHOs via nitrite, which reduced the need for both oxygen and carbon in nitrogen removal. Moreover, the microbial and DO profiles within the floc indicated a distinct stratification, with decreasing DO and OHOs, and increasing PAOs towards the inner layer. This study demonstrates a successful floc model that can be used to investigate and design SNDPR for scientific and practical purposes.

Evaluation of various carbon sources on ammonium assimilation and denitrifying phosphorus removal in a modified anaerobic-anoxic-oxic process from low-strength wastewater

A pilot-scale continuous-flow modified anaerobic-anoxic-oxic (MAAO) process examined the impact of external carbon sources (acetate, glucose, acetate/propionate) on ammonium assimilation, denitrifying phosphorus removal (DPR), and microbial community. Acetate exhibited superior efficacy in promoting the combined process of ammonia assimilation and DPR, enhancing both to 50.0 % and 60.0 %, respectively. Proteobacteria and Bacteroidota facilitated ammonium assimilation, while denitrifying phosphorus-accumulating organisms (DPAOs) played a key role in nitrogen (N) and phosphorus (P) removal. Denitrifying glycogen-accumulating organisms (DGAOs) aided N removal in the anoxic zone, ensuring stable N and P removal and recovery. Acetate/propionate significantly enhanced DPR (77.7 %) and endogenous denitrification (37.9 %). Glucose favored heterotrophic denitrification (29.6 %) but had minimal impact on ammonium assimilation. These findings provide valuable insights for wastewater treatment plants (WWTPs) seeking efficient N and P removal and recovery from low-strength wastewater.

Influence of decreasing temperature on aerobic granular sludge - microbial community dynamics and treatment performance

Municipal wastewater in temperate climates is characterized by seasonal temperature changes. Temperature is a determining factor for biological processes, but the impact of gradually decreasing temperature on aerobic granular sludge (AGS) has been largely unexplored. In this study, the influence of decreasing temperature from 20 °C to 6 °C on AGS was investigated at rates of 0.5 and 1 °C per week. Temperature was a major driver for microbial community change, where the community response could be divided into three main subclusters. Strains within the guilds of ammonium- and nitrite-oxidising bacteria (AOB and NOB) as well as polyphosphate- and glycogen-accumulating organisms (PAOs and GAOs) grouped in different subclusters, indicating variable responses among and between the guilds. The phosphorous removal rate was sufficient for complete removal at all temperatures, presumably due to functional redundancy within the PAOs. The nitrification rate was, however, seriously impaired below 13 °C, despite diversity within AOB and NOB.

Metabolic shifts in glycogen-accumulating organisms: Exploring carbon and phosphorus dynamics under two distinct phosphorus limitation modes

Glycogen-accumulating organisms (GAOs) are recognized as competitors of phosphorus-accumulating organisms (PAOs) in enhanced biological phosphorus removal (EBPR) systems. However, different types of GAOs exhibit unique metabolic traits in different environments. To characterize the metabolic activity of GAOs, two distinct modes were constructed. Long-term operation with the mode of anaerobic post-drainage demonstrated the successful establishment of a rapid and enduring GAOs system dominated by Candidatus Contendobacter community. Acetate was completely consumed within 60 min of the anaerobic phase. Concurrently, glycogen levels decreased by 0.062 g Glycogen/g SS, while intracellular carbon storage of poly-hydroxybutyrate (PHB) and poly-hydroxyvalerate (PHV) increased by 0.027 g PHB/g SS and 0.007 g PHV/g SS, respectively. Various concentrations of phosphorus loads illustrated the resistance of the system to high phosphorus loads, while maintaining glycogen accumulating metabolism (GAM). Under complete phosphorus limitation, no correlation between glycogen and acetate was observed during the initial 140 d. The addition of 1 mg/L phosphate to the influent preserved the typical GAM characteristics. Moreover, the pathway involved in glycogen conversion to trehalose may be pivotal in enhancing anaerobic glycogen degradation. The complete tricarboxylic acid cycle ensured efficient energy production, thereby sustaining the foundational metabolism. Notably, sucC and mdh were critical regulators of energy flux. This study provides a distinctive metabolic profile of a robust and resilient GAOs model with Ca. Contendobacter.

PHA from acetic acid: Manipulating microbial production of polyhydroxyalkanoates for use in steel coatings

This research sought to determine the impact of various culture parameters on the composition of polyhydroxyalkanoate (PHA), which was intended for use in steel coating. PHAs are biodegradable biopolymers that can be made by glycogen accumulating organisms (GAOs), which produce two types of monomers, 3-hydroxybutyrate (HB), and 3-hydroxyvalerate (HV). Several different culture parameters (acetate concentration, nitrogen and phosphate availability, temperature and pH) were varied and their effect on the PHA composition was measured. The results showed that the HB relative content was lower at acetate concentrations higher than 6 g/L and pH 6.5 and 7. The HV content increased from 26 % to 43 % for 18 °C and 28 °C, respectively. Nitrogen concentration did not influence PHA composition whereas 100 mg/L of phosphate induced a HV content equal to 47 %. A positive Pearson correlation between HB content and PHA yield was found in this research. This study showed how the culture conditions might be varied to change the PHA composition to serve a specific function, like replacing steel coating with biodegradable polymers.

Blind spots of universal primers and specific FISH probes for functional microbe and community characterization in EBPR systems.

Fluorescence in situ hybridization (FISH) and 16S rRNA gene amplicon sequencing are commonly used for microbial ecological analyses in biological enhanced phosphorus removal (EBPR) systems, the successful application of which was governed by the oligonucleotides used. We performed a systemic evaluation of commonly used probes/primers for known polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs). Most FISH probes showed blind spots and covered nontarget bacterial groups. Ca. Competibacter probes showed promising coverage and specificity. Those for Ca. Accumulibacter are desirable in coverage but targeted out-group bacteria, including Ca. Competibacter, Thauera, Dechlorosoma, and some polyphosphate-accumulating Cyanobacteria. Defluviicoccus probes are good in specificity but poor in coverage. Probes targeting Tetrasphaera or Dechloromonas showed low coverage and specificity. Specifically, DEMEF455, Bet135, and Dech453 for Dechloromonas covered Ca. Accumulibacter. Special attentions are needed when using these probes to resolve the PAO/GAO phenotype of Dechloromonas. Most species-specific probes for Ca. Accumulibacter, Ca. Lutibacillus, Ca. Phosphoribacter, and Tetrasphaera are highly specific. Overall, 1.4% Ca. Accumulibacter, 9.6% Ca. Competibacter, 43.3% Defluviicoccus, and 54.0% Dechloromonas in the MiDAS database were not covered by existing FISH probes. Different 16S rRNA amplicon primer sets showed distinct coverage of known PAOs and GAOs. None of them covered all members. Overall, 520F-802R and 515F-926R showed the most balanced coverage. All primers showed extremely low coverage of Microlunatus (<36.0%), implying their probably overlooked roles in EBPR systems. A clear understanding of the strength and weaknesses of each probe and primer set is a premise for rational evaluation and interpretation of obtained community results.

Bacterial community and filamentous population of industrial wastewater treatment plants in Belgium.

The discharge of industrial water requires the removal of its pollutants, where biological wastewater treatment plants (WWTPs) are the most used systems. Biological WWTPs make use of activated sludge (AS), where bacteria are responsible for the removal of pollutants. However, our knowledge of the microbial communities of industrial plants is limited. Understanding the microbial population is essential to provide solutions to industrial problems such as bulking. The aim of this study was to identify at a high taxonomic resolution the bacterial population of 29 industrial WWTPs using 16S rRNA amplicon sequencing. Our results revealed that the main functional groups were dominated by Thauera and Zoogloea within denitrifiers, Dechloromonas in phosphate-accumulating organisms, and Defluviicoccus in glycogen-accumulating organisms. The activated sludge characterization indicated that 59% of the industrial plants suffered from bulking sludge, with DSVI values of up to 448 mL g-1. From the bulking cases, 72% corresponded to filamentous bulking with Thiothrix as the most abundant filament; meanwhile, the other 28% corresponded to viscous bulking sludge in which Zoogloea was the most abundant genus. Furthermore, the bacterial population did not share a core of taxa across all industrial plants. However, 20 genera were present in at least 50% of the plants comprising the general core, including Thauera, Ca. Competibacter, and several undescribed microorganisms. Moreover, statistical analysis revealed that wastewater salinity strongly affected the microbial richness of the industrial plants. The bacterial population across industrial plants differed considerably from each other, resulting in unique microbial communities that are attributed to the specificity of their wastewaters. KEY POINTS: • The general core taxa of industrial plants were mostly made up of undescribed bacterial genera. • Filamentous bacteria constituted on average 4.1% read abundance of the industrial WWTPs. • Viscous bulking remains a significant type of bulking within industrial WWTPs.

Side-Stream Enhanced Biological Phosphorus Removal (S2EBPR) enables effective phosphorus removal in a pilot-scale A-B stage shortcut nitrogen removal system for mainstream municipal wastewater treatment

While the adsorption/bio-oxidation (A/B) process has been widely studied for carbon capture and shortcut nitrogen (N) removal, its integration with enhanced biological phosphorus (P) removal (EBPR) has been considered challenging and thus unexplored. Here, full-scale pilot testing with an integrated system combining A-stage high-rate activated sludge with B-stage partial (de)nitrification/anammox and side-stream EBPR (HRAS-P(D)N/A-S2EBPR) was conducted treating real municipal wastewater. The results demonstrated that, despite the relatively low influent carbon load, the B-stage P(D)N-S2EBPR system could achieve effective P removal performance, with the carbon supplement and redirection of the A-stage sludge fermentate to the S2EBPR. The novel process configuration design enabled a system shift in carbon flux and distribution for efficient EBPR, and provided unique selective factors for ecological niche partitioning among different key functionally relevant microorganisms including polyphosphate accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs). The combined nitrite from B-stage to S2EBPR and aerobic-anoxic conditions in our HRAS-P(D)N/A-S2EBPR system promoted DPAOs for simultaneous internal carbon-driven denitrification via nitrite and P removal. 16S rRNA gene-based oligotyping analysis revealed high phylogenetic microdiversity within the Accumulibacter population and discovered coexistence of certain oligotypes of Accumulibacter and Competibacter correlated with efficient P removal. Single-cell Raman micro-spectroscopy-based phenotypic profiling showed high phenotypic microdiversity in the active PAO community and the involvement of unidentified PAOs and internal carbon-accumulating organisms that potentially played an important role in system performance. This is the first pilot study to demonstrate that the P(D)N-S2EBPR system could achieve shortcut N removal and influent carbon-independent EBPR simultaneously, and the results provided insights into the effects of incorporating S2EBPR into A/B process on metabolic activities, microbial ecology, and resulted system performance.

Optimizing sludge retention time for sustainable photo-enhanced biological phosphorus removal systems: Insights into nutrient fate, microbial community, and bacterial phototolerance

Photo-enhanced Biological Phosphorus Removal (PEBPR) systems, promising wastewater treatment technology, offer efficient phosphorus removal without external oxygen. However, comprehending the impact of sludge retention time (SRT) on the system is crucial for successful implementation. This study investigated the SRT effect on nutrient fate, microbial community, and bacterial phototolerance in PEBPR systems. PEBPR systems exhibited good bacterial phototolerance at SRT of 10, 15, and 20 d, with optimal phosphorus-accumulation metabolism observed at SRT of 10 and 15d. However, at SRT of 5d, increased light sensitivity and glycogen-accumulating organisms (GAOs) growth resulted in poor P removal (71.9%). Accumulibacter-IIC were the dominant P accumulating organisms (PAOs) at SRT of 10, 15, and 20 d. Accumulibacter-I, IIC and IIF were the major PAOs at SRT of 5 d. The decrease in SRT promoted the microalgal population diversity, and Dictyosphaerium and Chlorella were the major microalgal species in this study. Flow cytometry results revealed high light intensity triggered intracellular Fe2+ efflux, limiting translation activity and metabolism. Moreover, PAOs had lower phototolerance than GAOs due to Poly-P bound intracellular Mg2+ affecting enzyme activity. This study provides an in-depth understanding of PEBPR systems operation strategy toward environmentally sustainable wastewater treatment.

New insight into the granule formation in the reactor for enhanced biological phosphorus removal.

While granulated activated sludge exhibits high productivity, the processes of granule formation are incompletely studied. The processes of granule formation and succession of communities were investigated in a laboratory sequencing batch reactor (SBR) under conditions for enhanced biological phosphorus removal (EBPR) using microbiological and molecular techniques. Active consumption of acetate, primarily by the phosphate-accumulating organisms (PAO), commenced at day 150 of cultivation. This was indicated by the high ratio of molar P-released/acetate uptake (0.73-0.77 P-mol/C-mol), characteristic of PAO. During this period, two types of granule-like aggregates formed spontaneously out of the activated sludge flocs. The aggregates differed in morphology and microbial taxonomic composition. While both aggregate types contained phosphorus-enriched bacterial cells, PAO prevailed in those of morphotype I, and glycogen-accumulating organisms (GAOs) were predominant in the aggregates of morphotype II. After 250 days, the elimination of the morphotype II aggregates from the reactor was observed. The subsequent selection of the community was associated with the development of the morphotype I aggregates, in which the relative abundance of PAO increased significantly, resulting in higher efficiency of phosphorus removal. Metagenomic analysis revealed a predominance of the organisms closely related to Candidatus Accumulibacter IС and IIС and of Ca. Accumulibacter IIB among the PAO. Based on the content of the genes of the key metabolic pathways, the genomes of potential PAO belonging to the genera Amaricoccus, Azonexus, Thauera, Zoogloea, Pinisolibacter, and Siculibacillus were selected. The patterns of physicochemical processes and the microbiome structure associated with granule formation and succession of the microbial communities were revealed.

Enhanced phosphorus removal from biological secondary effluent using denitrifying phosphorus-removal granular sludge

With the phosphorus discharge standard getting more stringent, it is imperative to enhance phosphorus removal from biological secondary effluent (BSE) of wastewater treatment plants (WWTPs). This study proposed a semi-continuous process for enhanced phosphorus removal from BSE using denitrifying phosphorus-removal granular sludge (DPGS) in a fluidized reactor operating under the alternative anaerobic/anoxic mode. This study investigated the effects of hydraulic retention time (HRT) and treating volume of BSE on the enhanced phosphorus removal performance of this process. At the appropriate HRT and treating volume of BSE, this process reduced the total phosphorus of practical BSE from 0.54 to 0.14 mg/L with an average removal efficiency of 74.5 % and removed 2.1 mg/L nitrate in 95 days. Removed phosphorus from practical BSE was concentrated into a small volume of water via the anaerobic phosphorus release and anoxic phosphorus uptake of denitrifying phosphorus-accumulating organisms (DPAOs) in DPGS, which was conducive for phosphorus recovery via chemical precipitation. The metagenomic analysis reveals that the relative abundance of DPAOs in DPGS increased while that of denitrifying glycogen-accumulating organisms decreased during 95 days of operation. This study provides a more cost-efficient and eco-friendly process for WWTPs to enhance phosphorus removal from BSE and phosphorus recycling.

Achieving rapid endogenous partial denitrification by regulating competition and cooperation between glycogen accumulating organisms and phosphorus accumulating organisms from conventional activated sludge

In anaerobic/aerobic/anoxic (A/O/A) process, endogenous denitrification (ED) is critically important, and achieving steady endogenous partial denitrification (EdPD) is crucial to carbon saving and anammox application. In this study, EdPD was rapidly realized from conventional activated sludge by expelling phosphorus accumulating organisms (PAOs) in anaerobic/anoxic (A/A) mode during 40 days, with nitrite transformation rate (NTR) surging to 82.8 % from 29.4 %. Competibacter was the prime EdPD-fulfilling bacterium, soaring to 28.9 % from 0.5 % in phase II. Afterwards, balance of high NTR and phosphorus removal efficiency (PRE) were attained by well regulating competition and cooperation between PAOs and glycogen accumulating organisms (GAOs) in A/O/A mode, when the Competibacter (21.7 %) and Accumulibacter (7.3 %, mainly Acc_IIC and Acc_IIF) were in dominant position with balance. The PRE recovered to 88.6 % and NTR remained 67.7 %. Great balance of GAOs and PAOs contributed to advanced nitrogen removal by anammox.

Long-term effects of thiosulfate on the competition between sulfur-mediated bacteria and glycogen accumulating organisms in sulfate-rich carbon-deficient wastewater

Sewage nutrient (e.g., nitrogen and phosphorus) biological removal performance is often limited by the deficient carbon source and undesirable glycogen accumulating organisms (GAOs), even in sulfate-containing wastewater. Thiosulfate (S2O32−) as a bioavailable, environmentally-benign, metastable and cost-effective agent has been regarded as electron carriers that induces high sulfur-mediated bacterial activity for nutrient removal from wastewater. In this study, the long-term effects of thiosulfate on the competition between sulfur-mediated bacteria (SMB, including sulfur-reducing bacteria (SRB) and sulfur-oxidizing bacteria (SOB)) and GAOs were explored to further close the gap of our knowledge on the control of GAOs under carbon deficient wastewater. Three reactors were continuously operated for over 100 days and were fed with 200 mg acetate-COD/L and 20 (R1), 50 (R2) and 80 (R3) mg S/L thiosulfate respectively. The results revealed that adding thiosulfate at the beginning of the anoxic phase promoted sulfur metabolism and increased the proliferation of SRB (mainly Desulfobacter) and SOB (mainly Chromatiaceae). Correspondingly, the relative abundance of GAOs (mainly Candidatus_Competibacter) decreased. After the carbon source was reduced, the abundance of GAOs increased and the competitive activity of SRB was weakened, resulting in the reduced sulfate reduction, which could be attributed to the fact that GAOs had a higher carbon source competitiveness than SRB under low carbon source conditions. While SOB maintained a high abundance due to the addition of thiosulfate as an additional electron donor, which enhanced the denitrification efficiency. Additionally, the dominant SOB shifted from Thiobacillus to Chromatiaceae during the long-term operation, indicating that Chromatiaceae had a higher competitive advantage for reduced sulfur (e.g., S2O32−, Polysulfide (Poly-S)) and nitrate compared to Thiobacillus. Furthermore, microbial functional genes revealed that S metabolism was enhanced during long-term operation. The potential mechanism and optimization strategy regarding the competition between sulfur-mediated bacteria and GAOs were revealed.

Performance and mechanism of glycerol-driven denitrifying phosphorus removal from low organic matter sewage

The performance and mechanism of the glycerol-driven denitrifying phosphorus removal (DPR) process were investigated in low organic matter wastewater treatment using the modified anaerobic-anoxic–oxic (MAAO) system. The results revealed that denitrifying bacteria preferentially utilized glycerol, reducing nitrate interference on anaerobic phosphate release. Fermentation bacteria converted excess glycerol into available carbon sources, which were utilized by denitrifying phosphorus-accumulating organisms (DPAOs). Optimize glycerol dosage (calculated in chemical oxygen demand) could be estimated based on 6 times the effluent NO3−-N of the anoxic zone. As glycerol dosage increased, the relative abundance of fermentation bacteria surged from 8.2% to 17.7%, subsequently boosting the DPR rate from 34.6% to 77.2%. Notably, denitrifying glycogen-accumulating organisms (DGAOs) decreased from 0.5% to 0.2% but remained instrumental in nitrogen removal. The collaborative actions of fermentation bacteria, DPAOs, and DGAOs were vital in upholding the stability of nutrient removal in the glycerol-driven DPR process.

The successful application of light to the system of simultaneous nitrification/endogenous denitrification and phosphorus removal: Promotion of partial nitrification and glycogen accumulation metabolism

Partial nitrification (PN) and high glycogen accumulating metabolism (GAM) activity are the basis for efficient nitrogen (N) and phosphorus (P) removal in simultaneous nitrification endogenous denitrification and phosphorus removal (SNDPR) systems. However, achieving these processes in practical operations is challenging. This study proposes that light irradiation is a novel strategy to enhance the nutrient removal performance of the SNDPR system with low carbon to nitrogen ratios (C/N of 3.3–4.1) domestic wastewater. Light energy densities (Es) of 55–135 J/g VSS were found to promote the activity of ammonia-oxidizing bacteria (AOB) and GAM, while inhibiting the activity of nitrite-oxidizing bacteria (NOB) and polyphosphate accumulating metabolism (PAM). Long-term exposure to different light patterns at Es of 55–135 J/g VSS revealed that continuous light rapidly achieved PN by inhibiting NOB activity and promoted the growth of glycogen accumulating organisms (GAOs), allowing the removal of above 82 % N and below 80 % P. Intermittent light maintained stable PN by inhibiting the activity and growth of NOB and promoted the growth of polyphosphate accumulating organisms (PAOs) with high GAM activity (Accmulibacer IIC-ii and IIC-iii), allowing the removal of above 82 % N and 95 % P. Flow cytometry and enzyme activity assays showed that light promoted GAM-related enzyme activity and the metabolic activity of partial Accmulibacer II over other endogenous denitrifying bacteria, while inhibiting NOB translation activity. These findings provide a new approach for enhancing nutrient removal, especially for achieving PN and promoting GAM activity, in SNDPR systems treating low C/N ratio domestic wastewater using light irradiation.