Autotrophic Nitrogen Removal Research Articles

Achieving efficient autotrophic nitrogen removal in anaerobic membrane bioreactor plus membrane aerated biofilm reactor by regulating nutrient ratios.

It is feasible to integrate an anaerobic membrane bioreactor with a membrane aerated biofilm reactor to efficiently implement the sulfate reduction, simultaneous nitrification and autotrophic denitrification process. However, the effect of parameters on nutrient removal and environmental impacts of the process are unclear. In this study, the reactor performance was mainly influenced by the chemical oxygen demand to sulfate (COD/S) ratio and the ammonium to sulfate (N/S) ratio in long-term operation. Significant models were developed to optimize the two factors using the response surface methodology. Under optimal conditions (COD/S ratio of 2.5 and N/S ratio of 0.3), the system could remove above 86 % COD, 99 % ammonium, and 92 % total inorganic nitrogen. Moreover, this process could reduce energy consumption by 30 % and global warming potential by 50 % compared with traditional anaerobic/oxic activated sludge process. These findings provide guidance for the application of this technology in sulfate-containing municipal sewage treatment.

Metatranscriptomics provides an in-depth perspective on the resistance and detoxification of anammox bacteria to dissolved oxygen in a pilot CANON process

In the completely autotrophic nitrogen removal over nitrite (CANON) process, the conflicting oxygen requirements of anammox and ammonium-oxidizing bacteria often lead to retardation in anammox activity. However, our study achieved stable nitrogen removal with a maximum capacity of 1096 g-N/m3/d in a 20 m3 CANON reactor under long-term intensive aeration. The anammox bacteria unusually distributed in the outer layer of the biofilm and demonstrated remarkable oxygen tolerance. Their activity only declined by 18.5 % under 2.0 mg/L of dissolved oxygen. When anammox bacteria encountered oxygen exposure, they adopted some strategies. Metatranscriptomics revealed that Candidatus Kuenenia, the dominant anammox species in our system, downregulated its gene expressions involved in carbon metabolism and oxidative phosphorylation. This may reduce electron leakage that combines with O2, thereby minimizing the generation of reactive oxygen species (ROS). By contrast, the secretion of extracellular proteins and conversion of O2·− were upregulated to eliminate ROS promptly. This behavior endowed Ca. Kuenenia with a unique oxygen detoxification pathway: O2·− were initially converted to H2O2 by superoxide dismutase SOD2 and superoxide reductase dfx (major role), followed by reduction to H2O via non-heme chloroperoxidase cpo (a newly recognized mechanism in the oxygen detoxification of anammox) and catalase katE. These results expanded the current knowledge of anammox alleviating oxidative stress.

Typical Heterotrophic and Autotrophic Nitrogen Removal Process Coupled with Membrane Bioreactor: Comparison of Fouling Behavior and Characterization.

There is limited research on the relationship between membrane fouling and microbial metabolites in the nitrogen removal process coupled with membrane bioreactors (MBRs). In this study, we compared anoxic-oxic (AO) and partial nitritation-anammox (PNA), which were selected as representative heterotrophic and autotrophic biological nitrogen removal-coupled MBR processes for their fouling behavior. At the same nitrogen loading rate of 100 mg/L and mixed liquor suspended solids (MLSS) concentration of 4000 mg/L, PNA-MBR exhibited more severe membrane fouling compared to AO-MBR, as evidenced by monitoring changes in transmembrane pressure (TMP). In the autotrophic nitrogen removal process, without added organic carbon, the supernatant of PNA-MBR had higher concentrations of protein, polysaccharides, and low-molecular-weight humic substances, leading to a rapid flux decline. Extracellular polymeric substances (EPS) extracted from suspended sludge and cake sludge in PNA-MBR also contributed to more severe membrane fouling than in AO-MBR. The EPS subfractions of PNA-MBR exhibited looser secondary structures in protein and stronger surface hydrophobicity, particularly in the cake sludge, which contained higher contents of humic substances with lower molecular weights. The higher abundances of Candidatus Brocadia and Chloroflexi in PNA-MBR could lead to the production of more hydrophobic organics and humic substances. Hydrophobic metabolism products as well as anammox bacteria were deposited on the hydrophobic membrane surface and formed serious fouling. Therefore, hydrophilic membrane modification is more urgently needed to mitigate membrane fouling when running PNA-MBR than AO-MBR.

Partitioned granular sludge coupling with membrane-aerated biofilm reactor for efficient autotrophic nitrogen removal

The partial nitritation-anammox process based on a membrane-aerated biofilm reactor (MABR) faces several challenges, such as difficulty in suppressing nitrite-oxidizing bacteria (NOB), excessive effluent nitrate, and ineffective synergy between denitrification and anammox bacteria. Therefore, a novel partitioned granular sludge coupling with MABR (G-MABR) was constructed. The chemical oxygen demand (COD) and nitrogen removal efficiency were 88.8 ± 1.8 %–92.6 ± 1.2 % and 88.8 ± 1.5 %–93.6 ± 0.7 %, respectively. The COD was mainly lowered in the lower granular sludge-zone, while nitrogen was removed in the upper MABR-zone. NOB was significantly suppressed in the MABR-zone due to competition for substrate with denitrifying bacteria and anammox bacteria. This partitioned configuration reduced the C/N ratio in the MABR-zone, thus facilitating autotrophic nitrogen removal. Both partial nitrification and denitrification provided nitrite for anammox bacteria in granular sludge, whereas partial nitrification mainly supplied nitrite to the anammox bacteria in membrane biofilms.

Comammox rather than AOB dominated the efficient autotrophic nitrification-denitrification process in an extremely oxygen-limited environment

The discovery of complete ammonia oxidizer (comammox) has challenged the traditional understanding of the two-step nitrification process. However, their functions in the oxygen-limited autotrophic nitrification-denitrification (OLAND) process remain unclear. In this study, OLAND was achieved using comammox-dominated nitrifying bacteria in an extremely oxygen-limited environment with a dissolved oxygen concentrations of 0.05 mg/L. The ammonia removal efficiency exceeded 97 %, and the total nitrogen removal efficiency reached 71 % when sodium bicarbonate was used as the carbon source. The pseudo-first- and second-order models were found to best fit the ammonia removal processes under low and high loads, respectively, suggesting distinct ammonia removal pathways. Full-length 16S rRNA gene sequencing and metagenomic results revealed that comammox-dominated under different oxygen levels, in conjunction with anammox and heterotrophic denitrifiers. The abundance of enzymes involved in energy metabolism indicates the coexistence of anammox and autotrophic nitrification-heterotrophic denitrification pathways. The binning results showed that comammox bacteria engaged in horizontal gene transfer with nitrifiers, anammox bacteria, and denitrifiers to adapt to an obligate environments. Therefore, this study demonstrated that comammox, anammox, and heterotrophic denitrifiers play important roles in the OLAND process and provide a reference for further reducing aeration energy in the autotrophic nitrogen removal process.

Autotrophic biological nitrogen removal in anammox system at elevated temperature: EPS regulation and metagenomic analysis

Carbon reduction in wastewater treatment industry has been attracted global attention. Anaerobic ammonium oxidation (anammox) process is known as a promising and energy-efficient technology. In this study, the effects of elevated temperature on anammox system were thoroughly investigated. Analyses of effluent and microbial community indicated that long-term exposure to 45℃ led to system collapse and a huge shift in community structure. Network analysis revealed that the sudden drop in abundance of AnAOB at 45℃ (from 36.19 % to 10.71 %) made them uncompetitive with heterotrophic bacteria. The decrease of EPS concentration (a minimum of 7.22 mg·g−1-VSS at 45℃) combined with the enhancement of EPS metabolism at higher temperature suggested a shift in the role of EPS from protective function at 35℃ to utilization as a carbon source for heterotrophic bacteria at higher temperatures. Through constructing the cross-feeding relationships associated with amino acids, polysaccharides, and cofactors, we found the dependence of heterotrophs on AnAOB became weaker at high temperature, and the disturbances in interspecific relationships might be responsible for the collapse of the anammox system. This work presents significant insights into the engineering of anammox process under high temperature.

Integration of mixotrophic denitrification and anammox in Thiothrix-hydroxyapatite coupled particles treating municipal wastewater against low temperature and organic load shocks

Sulfur-driven autotrophic denitrification coupled with anammox (SPDA) is a promising process for autotrophic nitrogen removal, but autotrophic microorganisms are challenging to retain in wastewater treatment units. In this study, a single-stage mixotrophic denitrification coupled with anammox (MDA) process was established in an up-flow anaerobic sludge bed reactor with Thiothrix-hydroxyapatite coupled particles. The advancement of the sludge granulation process effectively strengthened the resistance of the MDA system to environmental stresses, and the SPDA-dominated nitrogen removal process achieved a total nitrogen removal efficiency of 88.6 %, a nitrogen removal rate of 0.432 KgN/m3/d at temperatures as low as 12.8 °C and an organic loading rate of 0.63 KgCOD/m3/d. Interestingly, the ecological niche integration of sulfur oxidizing bacteria (SOB), anammox bacteria and heterotrophic denitrifying bacteria in the granular sludge was observed during long-term operation. The FISH-CLSM results revealed the spatial distribution of functional microorganisms in the granular sludge and the potential pollutant transformation mechanism. The aggressive growth of SOB dominated by Thiothrix constructed a biological wall at the periphery of the granules, which ensured their priority for nitrate acquisition, supplied nitrite to the internal anammox bacteria, and acted as an important barrier against the shock of organic loads. Overall, this study presents a novel granulation approach for the biological treatment of sulfur-containing wastewater, highlighting the efficacy of functional integration within Thiothrix-hydroxyapatite coupled particles as an effective strategy to resist environmental stress.

Rapid Start-up of Continuous Autotrophic Nitrogen Removal Reactor by Hybrid-inoculating PN and PN/A Granular Sludges

The rapid cultivation of partial nitritation/ANAMMOX （PN/A） granular sludge in a continuous-flow mode is one of the key technologies for efficient biological nitrogen removal in domestic wastewater treatment. Compared with that in PN/A granular sludge, PN granular sludge demonstrates a shorter incubation period and suitability for batch culture. It is also a good carrier for enriching ANAMMOX （AMX） bacteria. In this study, we established a continuous-flow autotrophic nitrogen removal process in three continuously stirred tank reactors （CSTR） （R1-R3） by hybrid-inoculating PN/A and PN granular sludge at the mass ratios of 3∶1, 1∶1, and 1∶3, respectively. By implementing high ammonium nitrogen loading and short hydraulic retention time, continuous autotrophic nitrogen removal processes were successfully started up in the three CSTRs. The results showed that compared with that of R1 and R2, R3 had a longer start-up time but a similar steady-state nitrogen removal performance. The total nitrogen removal load of R3 could be more than 2.6 kg·（m3·d）-1. Intriguingly, the inoculated PN granular sludge served as a precursor for PN/A granular sludge cultivation. This approach facilitated the enrichment of anaerobic ammonia-oxidizing bacteria （AMX） by introducing abundant ammonium-oxidizing bacteria （AOB） and nitrite nitrogen substrates into the CSTR. According to the results of high-throughput sequencing, the microbial abundance and diversity of the mature granules in R1-R3 were significantly higher than those of the inoculation sludge. AOB （genus Nitrosomonas）, AMX （genera Candidatus Kuenenia and Candidatus Brocadia）, and symbiotic heterotrophs, such as Chloroflexi, Bacteroidetes, and Chlorobi, drove the autotrophic nitrogen removal process and maintained the stability of the granular structure. In summary, a novel start-up strategy of hybrid-inoculating granular sludge was provided for a continuous-flow autotrophic nitrogen removal in engineering application.

External electron donor and transporter of iron triggers the paths to enhancing collaboration of anammox and denitrification in granular sludge: Bacterial resilience and synergetic promotion mechanism

Autotrophic nitrogen removal via anaerobic ammonium oxidation (anammox) technology currently faces significant challenges in treating low-strength wastewater due to inefficient retention of bacteria and unstable activity under fluctuating conditions. This study demonstrates enhanced resilience and stability of anammox coupled with partial denitrification (PD) in granular sludge, assisted by ferric iron (Fe(III)) and nano zero-valent iron (nZVI). The systems treated wastewater with low concentrations of ammonia (NH4+-N of 50 ∼ 80 mg/L) and nitrate (NO3−-N of 60 ∼ 100 mg/L). Fe(III) and nZVI were intermittently added to two PD coupled with anammox (PD/A) systems, which were subjected to increasing nitrogen loading rates (NLRs) and decreasing temperature. Remarkably, both Fe(III)- and nZVI-assisted systems achieved satisfactory total nitrogen (TN) removal efficiencies of 98.1 % and 96.6 %, respectively, even as the temperature dropped from 28.5℃ to 13.4℃ and the NLR increased from 0.22 to 0.72 kgN/m3/d. Significantly, these two systems exhibited pronounced resilience under the shocking effect of excessive organic load, extremely low pH, and high salinity. The presence of iron, acting as an accelerated electron transporter, enhanced anammox activity and microbial NH4+-N oxidation. Additionally, iron reduced the requirement for an organic carbon source, further stimulating the competitiveness of anammox for nitrite (NO2−-N) against denitrification. Interestingly, metagenomic analysis revealed that the functional bacteria Thauera, responsible for PD, employed versatile metabolic pathways for NO3−-N reduction to NO2−-N using Fe(II) as an electron donor. Overall, this study provides new insights into microbial interactions and versatile metabolism in iron-assisted anammox systems, contributing to more energy-efficient wastewater treatment.

Implementing a completely autotrophic nitrogen removal over nitrite process using a novel umbrella basalt fiber carrier.

The completely autotrophic nitrogen removal over nitrite (CANON) process is significantly hindered by prolonged start-up periods and unstable nitrogen removal efficiency. In this study, a novel umbrella basalt fiber (BF) carrier with good biological affinity and adsorption performance was used to initiate the CANON process. The CANON process was initiated on day 64 in a sequencing batch reactor equipped with umbrella BF carriers. During this period, the influent NH4+-N concentration gradually increased from 100 to 200 mg·L-1, and the dissolved oxygen was controlled below 0.8 mg L-1. Consequently, an average ammonia nitrogen removal efficiency (ARE) and total nitrogen removal efficiency (TNRE) of ∼90 and 80% were achieved, respectively. After 130 days, ARE and TNRE remained stable at 92 and 81.1%, respectively. This indicates a reliable method for achieving rapid start-up and stable operation of the CANON process. Moreover, Candidatus Kuenenia and Candidatus Brocadia were identified as dominant anammox genera on the carrier. Nitrosomonas was the predominant genus among ammonia-oxidizing bacteria. Spatial differences were observed in the microbial population of umbrella BF carriers. This arrangement facilitated autotrophic nitrogen removal in a single reactor. This study indicates that the novel umbrella BF carrier is a highly suitable biocarrier for the CANON process.

Mechanism of stable autotrophic nitrogen removal and NOB inhibition in low-sludge PN-Anammox system under non-temperature control and low ammonia environment

PN/Anammox, as an economical and effective nitrogen removal process, is of great significance for reducing carbon emission in municipal wastewater treatment. However, municipal wastewater has mainstream characterized of low temperature and low NH4+-N concentration, and NOB activity is difficult to inhibit, so achieving a stable PN process is the biggest challenge in the application of PN/Anammox to mainstream wastewater. This study developed a method for stable and efficient PN/Anammox based on very low sludge volume (MLVSS = 65 mg/L). Even when the temperature dropped to 15℃ or there was exogenous nitrite-oxidizing bacteria (NOB) interference, the system still had good inhibition of NOB activity. The total stable operation time was 386 days, and the NO2–-N accumulation efficiency could reach more than 90 %. Ellin6067, Nitrosomonas and Candidatus_Brocadia were the dominant functional microorganisms in the system. Nitrospira was the dominant genera of NOB with very low relative abundance (<0.04 %). The mechanism of the long-term stable mainstream PN/Anammox system showed that low sludge content was beneficial to light irradiation on microorganisms in the reactor, while nxrA, the functional gene of NOB, had weak light resistance and oxidation resistance, which makes the system had strong inhibition effect on NOB. The construction of this new system which does not depend on activated sludge volume and complex NOB activity inhibition strategy can better promote the practical application of PN/Anammox under mainstream conditions.

Advancing autotrophic nitrogen removal in low C/N ratio wastewater: Innovative application of supercapacitor to enhance microbial electrolysis cells

This study presents a novel approach, the Supercapacitor Microbial Electrolysis Cell (SC-MEC), which utilizes a supercapacitor as an external power source to enhance the efficiency of autotrophic nitrogen removal in low C/N ratio wastewater. The results demonstrated that the SC-MEC system, operating under anaerobic conditions and devoid of any organic carbon source, exhibited exceptional performance in ammonia oxidation and total nitrogen (TN) removal when solely relying on ammonia nitrogen as the electron donor. Operating at a voltage of 1.8 V with a capacitance capacity of 30 F, ammonium oxidation rated up to 56.51 mg/L/day and TN removal rated up to 54.64 mg/L/day, in which 97% of ammonium nitrogen was converted to gaseous nitrogen. Furthermore, the charging and discharging process of supercapacitors autonomously regulated the bipolar potentials. Cyclic voltammetry (CV) analysis showed the significantly enhanced electrochemical activity of the SC-MEC system during the reaction process. Based on in-situ CV test results, it was inferred that this enhancement was associated with extracellular electron transfer mediators. The microbial community analysis revealed a process of synchronous nitrification and denitrification (SND) coupled with anammox, involving multiple genera, such as Candidatus Kuenenia, Nitrosomonas, Truepera, and Bosea. In conclusion, this study highlights the tremendous potential of SC-MEC in achieving efficient autotrophic nitrogen removal, offering more feasible and economical solutions for addressing low C/N water pollution issues.

Start-up of pilot-scale ANAMMOX reactor for low-carbon nitrogen removal from anaerobic digestion effluent of kitchen waste

The pilot-scale simultaneous denitrification and methanation (SDM)-partial nitrification (PN)-anaerobic ammonia oxidation (Anammox) system was designed to treat anaerobic digestion effluent of kitchen waste (ADE-KW). The SDM-PN was first started to avoid the inhibition of high-concentration pollutants. Subsequently, Anammox was coupled to realize autotrophic nitrogen removal. Shortcut nitrification–denitrification achieved by the SDM-PN. The NO2–-N accumulation (92 %) and NH4+-N conversion (60 %) were achieved by PN, and the removal of TN and COD from the SDM-PN was 70 % and 73 %, respectively. After coupling Anammox, the TN (95 %) was removed with a TN removal rate of 0.51 kg·m−3·d-1. Microbiological analyses showed a shift from dominance by Methanothermobacter to co-dominance by Methanothermobacter, Thermomonas, and Flavobacterium in SDM during the SDM-PN. While after coupling Anammox, Candidatus kuenenia was enriched in the Anammox zone, the SDM zone shifted back to being dominated by Methanothermobacter. Overall, this study provides new ideas for the treatment of ADE-KW.

Effect of suspended solids from anaerobic digested wastewater on performance and microbial community of autotrophic nitrogen removal process

Most anaerobically digested wastewaters have high concentrations of suspended solids, while little is known about whether they adversely affect the performance of the autotrophic nitrogen removal process. In this work, using air-lift internal circulation reactor we studied the effect of suspended solids on the autotrophic nitrogen removal process by adding 600–900 mg/L suspended solids from digested swine wastewater to the influent (R–SS). The results showed that during the formation of autotrophic nitrogen removal granular sludge, suspended solids containing Ca, Mg, Cu P, and Pb can enter the granular sludge, occupy the ecological niche of anammox bacteria, lead to loose internal structure, and damage the internal anoxic environment. It then reduces the settling performance in the R–SS, causes heavy wash-out of granular sludge from the R–SS reactor (2.5 times more than the control, R–CK), and finally reduces the maximum nitrogen removal rate of the autotrophic nitrogen removal process from 5.16 kg N/m3/d to 4.31 kg N/m3/d. Meanwhile, the metagenomic analysis showed that suspended solids reduce the relative abundance of reads from key nitrogen genes of anammox bacteria yet increases reads from both nitrite-oxidizing and denitrifying bacteria, especially nir and nor genes. Further analysis of 16 highly abundant genomes assembled via metagenomics showed that suspended solids decrease the relative abundance of Kuenenia_seed103, an anammox bacterium, from 13.36% (R–CK) to 11.03% (R–SS). Since suspended solids can act as a carbon source for denitrifiers to compete with nitrite or nitric oxide with anammox bacteria, the denitrifier community has progressed from a simple denitrifier consisting of only one or two denitrification steps (nar or nos) that acts as an ammonia and nitrite sink to a complete one that directly reduces nitrite to nitrogen gas. This study will lay a good foundation for its application in wastewater treatment.

Coupling Partial Nitritation, Anammox and n-DAMO in a membrane aerated biofilm reactor for simultaneous dissolved methane and nitrogen removal

Anaerobic technologies with downstream autotrophic nitrogen removal have been proposed to enhance bioenergy recovery and transform a wastewater treatment plant from an energy consumer to an energy exporter. However, approximately 20–50 % of the produced methane is dissolved in the anaerobically treated effluent and is easily stripped into the atmosphere in the downstream aerobic process, contributing to the release of greenhouse gas emissions. This study aims to develop a solution to beneficially utilize dissolved methane to support high-level nitrogen removal from anaerobically treated mainstream wastewater. A novel technology, integrating Partial Nitritation, Anammox and Methane-dependent nitrite/nitrate reduction (i.e. PNAM) was demonstrated in a membrane-aerated biofilm reactor (MABR). With the feeding of ∼50 mg NH4+-N/L and ∼20 mg/L dissolved methane at a hydraulic retention time of 15 h, around 90 % of nitrogen and ∼100 % of dissolved methane can be removed together in the MABR. Microbial community characterization revealed that ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), anammox bacteria, nitrite/nitrate-dependent anaerobic methane oxidation microorganisms (n-DAMO bacteria and archaea) and aerobic methanotrophs co-existed in the established biofilm. Batch tests confirmed the active microbial pathways and showed that AOB, anammox bacteria and n-DAMO microbes were jointly responsible for the nitrogen removal, and dissolved methane was mainly removed by the n-DAMO process, with aerobic methane oxidation making a minor contribution. In addition, the established system was robust against dynamic changes in influent composition. The study provides a promising technology for the simultaneous removal of dissolved methane and nitrogen from domestic wastewater, which can support the transformation of wastewater treatment from an energy- and carbon-intensive process, to one that is energy- and carbon-neutral.

S0-driven partial denitrification coupled with anammox (S0PDA) enables highly efficient autotrophic nitrogen removal from wastewater

This study proposed a novel strategy that integrates S0 particles (diameter: 2–3 mm) and granular sludge to establish S0-driven partial denitrification coupled with anammox (S0PDA) process for autotrophic nitrogen removal from NH4+- and NO3−-containing wastewaters. This process was evaluated using an up-flow anoxic sludge bed bioreactor, operating continuously for 240 days. The influent concentrations of NH4+ and NO3− were 29.9 ± 2.7 and 50.2 ± 2.7 mg-N/L, respectively. Throughout the operation, the hydraulic retention time was shortened from 4.0 h to 2.0 h, while the effluent concentrations of NH4+ and NO3− were maintained at a desirable level of 1.45–1.51 mg-N/L and 4.46–6.52 mg-N/L, respectively. Despite an autotrophic process, the nitrogen removal efficiency and rate reached up to 88.5 ± 2.0 % and 1.75 ± 0.07 kg-N/(m3·d), respectively, indicating the remarkable robustness of the S0PDA process. Autotrophic anammox and sulfur-oxidizing bacteria (Candidatus Brocadia and Thiobacillus) were the predominant bacterial genera involved in the S0PDA process. Candidatus Brocadia was primarily enriched in the granular sludge, with a relative abundance of 6.70 %. Thiobacillus occupied a unique niche on the S0 particles, with a relative abundance as high as 57.6 %, of which Thiobacillus thioparus with partial denitrification function (reducing NO3− to NO2− without further reduction to N2) accounted for 78.0 %. These findings challenge the stereotype of low efficiency in autotrophic nitrogen removal from wastewater, shedding fresh light on the applications of autotrophic processes.

Advanced nitrogen removal from municipal wastewater by autotrophy-heterotrophy coupled anammox system in a novel simultaneous microaerobic/limited-oxygen SBR: Interspecific correlation network

Nowadays, autotrophic nitrogen removal of mainstream municipal wastewater by anammox is highly promising. However, due to the lack of a long-term stable nitrite-oxidizing bacteria (NOB) inhibition strategy, the accumulation of NO3−-N byproducts severely affects the nitrogen removal efficiency (NRE). In this study, a simultaneous autotrophy-heterotrophy coupled anammox system was developed in a microaerobic/oxygen-limited SBR through the co-induction of the regulation of aeration intensity and limited organics to achieve the optimization of the mainstream Partial nitrification-anammox (PN/A) processes. The nitrogen removal system constructed by anammox and denitrification achieved the high NRE of 90.77 ± 1.82 %. Under the autotrophic-heterotrophic multiple induction of nitrite supply (PN, partial denitrification and endogenous partial denitrification), the contribution of the nitrogen removal by anammox increased to 57.93 %. The high-throughput sequencing analysis showed that the relative abundance of AnAOB increased from 0.40 % to 0.67 %. In-situ tests assay further demonstrated that the realization of multi-channel nitrite supply facilitated the enrichment of anammox bacteria (AnAOB). The endogenous denitrifying bacterial genus Candidatus (Ca.) Competibacter was significantly enriched in the system (7.98 %, P ≤ 0.001), which enhanced the deep removal of NOX−-N and organics. Correlation network analysis indicated that closer and more balanced interspecies interactions were the key to the high efficiency and stability of the autotrophy-heterotrophy coupled anammox system. Based on the nitrite recirculation and denitrification performance, the simultaneous autotrophy-heterotrophy coupled anammox system established by the novel SBR provides a potential strategy for mainstream anammox applications.

Formation of autotrophic nitrogen removal granular sludge driven by the dual-partition airlift internal circulation: Insights from performance assessment, community succession, and metabolic mechanism

Granular sludge has been recognized as an effective method for the application and industrialization of the anammox-based process due to its good biomass retention capacity and environmental tolerance. In this study, a one-stage autotrophic nitrogen removal (ANR) dual-partition system with airlift internal circulation was implemented for 320 days. A high nitrogen removal efficiency of 84.6% was obtained, while the nitrogen removal rate reached 1.28 g-N/L/d. ANR granular sludge dominated by Nitrosomonas and Candidatus Brocadia was successfully cultivated. Results showed that activity and abundance of functional flora first increased with granulation process, but eventually declined slightly when particle size exceeded the optimal range. Total anammox activity was observed to be significantly correlated with protein content (R2 = 0.9623) and nitrogen removal performance (R2 = 0.8796). Correlation network revealed that AnAOB had complex interactions with other bacteria, both synergy for nitrogen removal and competition for substrate. Changes in abundances of genes encoding the Carbohydrate Metabolism, Energy Metabolism, and Membrane Transport suggested energy production and material transfer were possibly blocked with further sludge granulation. Formation of ANR granular sludge promoted the interactions and metabolism of functional microorganisms, and the complex nitrogen metabolic pathways improved the performance stability. These results validated the feasibility of granule formation in the airlift dual-partition system and revealed the response of the ANR system to sludge granulation.

High ammonia loading rate and biofilm reattachment initiated partial nitrification and anammox in a membrane aerated biofilm reactor

Inhibition of nitrite oxidizing bacteria (NOB) is important for efficient wastewater treatment via anammox. Here, a novel strategy was demonstrated, which based on the detachment and reattachment of the biofilm to establish partial nitrification and anammox in a membrane aerated biofilm reactor (MABR). The results showed that using free ammonia as the sole inhibitor to start up partial nitrification took more time, and NOB acclimated to the free ammonia inhibition. Detaching the biofilm made it possible for free ammonia to contact NOB in deeper zones of the biofilm while at the same time washing NOB out of the reactor. Nitrite accumulation efficiency (NAE) increased to 70 % in 50 days when both high ammonia loading and biofilm reattachment were applied. With a short hydraulic retention time and high concentrations of ammonia and nitrite, anammox activity appeared in the reactor and resulted in a nitrogen removal efficiency of 60 %. Microbial community analysis indicated that Nitrosomonas, Nitrospira, and Candidatus Brocadia were the main functional taxa. Anaerobic ammonia oxidizing bacteria (AnAOB) proliferated within the biofilm, inhibiting NOB growth by competing for nitrite. Endogenous denitrifying bacteria also grew, coexisting with AnAOB. Predicted abundance of genes related to fermentation correlated with those involved in denitrification, indicating the possibility of heterotrophic denitrification. These findings help further our understanding of biofilm control-based strategies for NOB management and have implications for establishing autotrophic nitrogen removal using MABRs.

Cultivation of the coupling bacteria for simultaneous autotrophic nitrogen and sulfur removal and its remediation effect on the river sediment

The coupling bacteria for simultaneous autotrophic nitrogen and sulfur removal was cultivated by supplying sulfide to an Anammox biofilter treating low-strength wastewater. The results showed that sulfide-driven autotrophic denitrification (SAD) was successfully induced, and the total nitrogen (TN) removal increased from 83.0% to 94.4% when the sulfide concentration was increased to 25 mg L−1, with complete sulfide removal. Mass balance analysis suggested that 30.5% of the generated nitrate was reduced to nitrite while the rest was converted to nitrogen gas. The relative abundance of Anammox bacteria (Candidatus Kuenenia) increased from 9.16% to 18.8%, and the SAD bacteria (Thiobacillus) increased from 0.07% to 2.00%, both contributing collaboratively to nitrogen and sulfur removal. The coupling bacteria demonstrated a significant remediation effect on river sediment, increasing the TN removal activity by 10 times with a sludge addition ratio of 1:20. The results provide indications for enhancing the nitrogen and sulfur cycle in aquatic environments.