Nitrite-oxidizing Bacteria Research Articles

Response of ammonium transformation in bioanodes to potential regulation: Performance, electromicrobiome and implications

Understanding how potential regulation affects ammonium transformation in bioanodes is crucial for promoting their application. This study explored the performance, electrochemical properties, electromicrobiome of bioanodes across potentials from 0.0 V to 0.4 V vs. standard hydrogen electrode (SHE). Higher anode potentials enhanced the performance of electroactive biofilms and ammonium removal but suppressed nitrite oxidation while favoring dissimilatory nitrate reduction (DNRA), leading to increased nitrite accumulation. A reduction in nitrite-oxidizing bacteria (NOB) and an increase in DNRA-related genes resulted in an optimal nitrite-to-ammonium ratio of 1.32 for the Anammox process. Higher anodic potentials (0.3 and 0.4 V) were less effective for TN removal than lower potentials (0, 0.1, and 0.2 V), likely due to increased NOB and denitrification genes at lower potentials enhancing nitrite oxidation and denitrification. These findings indicate that regulating anodic potential effectively directs ammonium transformation in bioanodes, optimizing its conversion to N2 or nitrite.

Activity of novel virus families infecting soil nitrifiers is concomitant with host niche differentiation.

Chemolithoautotrophic nitrifiers are model groups for linking phylogeny, evolution, and ecophysiology. Ammonia-oxidising bacteria (AOB) typically dominate the first step of ammonia oxidation at high ammonium supply rates, ammonia-oxidising archaea (AOA) and complete ammonia-oxidising Nitrospira (comammox) are often active at lower supply rates or during AOB inactivity, and nitrite-oxidising bacteria (NOB) complete canonical nitrification. Soil virus communities are dynamic but contributions to functional processes are largely undetermined. In addition, characterising viruses infecting hosts with low relative abundance, such as nitrifiers, may be constrained by vast viral diversity, partial genome recovery, and difficulties in host linkage. Here, we describe a targeted incubation study that aimed to determine whether growth of different nitrifier groups in soil is associated with active virus populations and if process-focussed analyses facilitate characterisation of high-quality virus genomes. dsDNA viruses infecting different nitrifier groups were enriched in situ via differential host inhibition. Growth of each nitrifier group was consistent with predicted inhibition profiles and concomitant with the abundance of their viruses. These included 61 high-quality/complete virus genomes 35-173kb in length with minimal similarity to validated families. AOA viruses lacked ammonia monooxygenase sub-unit C (amoC) genes found in marine AOA viruses but some encoded AOA-specific multicopper oxidase type 1 (MCO1), previously implicated in copper acquisition, and suggesting a role in supporting energy metabolism of soil AOA. Findings demonstrate focussed incubation studies facilitate characterisation of active host-virus interactions associated with specific processes and viruses of soil AOA, AOB and NOB are dynamic and potentially influence nitrogen cycling processes.

Microalgae Enhance the Resistance of Pond-Dwelling Ammonia-Oxidizing Bacteria to Light Irradiation

Pond aquaculture is an important aquacultural model worldwide in which ammonia-oxidizing bacteria (AOB) are crucial for the removal of ammonia from water. The influence of light irradiation on AOB in an aquaculture pond was studied using artificial simulation wastewater under dark/light cycles of 24 h/0 h (L0), 12 h/12 h (L12), and 0 h/24 h (L24). The ammonia oxidation rates (AORs) in groups L0, L12, and L24 were 9.88 ± 0.19 mg h−1, 6.01 ± 0.32 mg h−1, and 1.85 ± 0.09 mg h−1, respectively. Long-term exposure to light had a serious impact on the AOR and decreased the abundance of Nitrosomonas spp. and their ammonia monooxygenase genes. To determine the protective effect of microalgae on AOB, different doses of freeze-dried Chlorella spp. powder were added to the nitrifying bacteria community. The photoinhibition rate of chlorophyll a (Chla) in the groups with 300 and 1300 µg L−1 of added Chlorella were 32.85% and 28.77%, respectively, while the Chla in the 2200 µg L−1 Chlorella-added group was only 0.01%, with no significant differences (p > 0.05) in AOR between the dark/light treatment subgroups. Fluorescence in situ hybridization showed that AOB, nitrite-oxidizing bacteria, and algae coexist and grow together without free AOB in the nitrifying bacterial community. It was suggested that microalgae enhance the resistance of AOB to light irradiation in a pond through the shading effect provided by algal chlorophyll and the close symbiotic relationship between microalgae and AOB.

Urban wastewater treatment at ambient conditions using microalgae-bacteria consortia in a membrane high-rate algal pond (MHRAP): The effect of hydraulic retention time and influent strength

The effects of hydraulic retention time (HRT) and influent wastewater strength on microalgae-bacteria performance were studied in two sets of experiments in a 6-day biomass retention time membrane high-rate algal pond (MHRAP) pilot plant to assess its pollutants removal potential. In the first set of experiments, the MHRAP was continuously fed with effluent from primary settling and operated at 6-, 4- and 2-days HRT and in the second set, the MHRAP was operated at 4-, 2-, and 1-day HRT using pre-treatment effluent. Results showed a complex interaction between algae-bacteria consortium, ambient conditions and MHRAP operation. In set 1, the highest total nitrogen (T-N) and total phosphorus (T-P) removal efficiencies were reached with 6-day HRT (23.6±0.6 % and 32±4 %, respectively), partially due to free ammonia nitrogen stripping and phosphorus precipitation, because of abiotic factors, which reduces the activity of nitrite-oxidizing bacteria and led to partial-nitrification. On the other hand, the 4 and 2-day HRT T-N and T-P removal efficiencies were reduced (below 10 % and 5 %, respectively) because the abiotic factors limited the physicochemical processes, and less biomass was taken up. In set 2, a suitable equilibrium between the microalgae and bacterial communities at 2-days HRT achieved high T-N (91±5 %) and T-P (71±8 %) removal efficiencies. Mass balances indicated that the nitrogen was mostly removed by nitrification-denitrification (87 % of T-N) and T-P predominant removal mechanism was precipitation. The COD removal efficiencies were higher than 86 % in both sets, since the particulate COD and colloids were removed by ultrafiltration. The obtained results showed that MHRAP technology would allow reducing COD and nutrient loads in the effluent under certain operating conditions, demonstrating the potential for resource recovery and wastewater treatment of microalgae-bacteria consortia.

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.

Predicting aeration time and nitrite accumulation rate variations for Partial Nitritation: A model incorporating nitrogen oxidation rate dynamics

This study aimed to develop a two-step nitrification model to predict variations in aeration time and nitrite accumulation rate (NAR) under fluctuating operational conditions in mainstream partial nitritation (PN) processes. Lab-scale sequencing batch reactors (SBRs) were used to evaluate the ammonia oxidation rate (AOR) and nitrite oxidation rate (NOR) under different solids retention times (SRT) (10, 15, 20, 30, and 50 days) and total volumetric nitrogen loadings (TVNL) (20–60 mg N/L per cycle). A static model was developed to predict consistent AOR and NOR values in the steady state, whereas a dynamic model was established to capture the growth dynamics of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) under unsteady-state conditions. The static model accurately predicted the AOR, NOR, and aeration time at steady state. The dynamic model quantified the relationship between specific growth rates (μ) and food-to-microorganism ratios (F/M) through exponential fitting, successfully capturing AOB and NOB growth dynamics. Validation experiments (SRT = 10 d, TVNL = 60 mg/L per cycle) demonstrated the ability of the dynamic model to predict trends in NAR and aeration time accurately. This study emphasizes the importance of accurately modeling AOR and NOR variations to predict aeration time and NAR, thereby providing valuable insights for aeration control and precise management of AOB and NOB populations in mainstream PN processes.

Effects of tourmaline on the establishment, idling, and recovery of partial nitrification: Mechanistic understanding and performance evaluation

The effects of different doses of tourmaline (Tm) on the partial nitrification (PN) system were investigated in this study. Tourmaline − Partial Nitrification sequential batch reactor (TmPNSBR) aimed to optimize the efficiency of PN for nitrogen removal in wastewater treatment. These results showed that Tm could reduce the time for PN establishment with a maximum reduction of 67.71% (Tm 8), maintained the stability of microbial community structure during the idle period, increased the ammonia oxidation rate, and promoted nitrite accumulation. Tm stimulated microorganisms to secrete more EPS (increase 7.92%–22.43% in phase I), and microorganisms adhere closely to the surface of Tm. Microbial community analysis showed that adding Tm could change the microbial community abundance and diversity, Tm increased the Ammonia-oxidizing bacteria (AOB) abundance (increase 7.92%–22.43% in phase I) and decreased the nitrite-oxidizing bacteria (NOB) abundance. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt2) predicted increased content of key enzymes associated with PN and electron transport-related complexes. This study delved into the potential of Tm to mitigate environmental pollution as an efficient and environmentally friendly new material that optimizes the PN process, thereby contributing to the energy-efficient treatment of sustainable wastewater.

Enhancing nitrogen removal by simultaneous nitritation and denitritation in a multi-cycle SBR with supplementation of solid carbon sources

Simultaneous nitritation and denitritation have the potential to significantly improve nitrogen removal in sewage treatment processes. However, their application in low-strength sewage treatment systems presents challenges. This study explored the impact of four solid carbon sources (SCSs) on N-removal via nitrite in a multi-cycle SBR with biocarriers. Results showed that both N-removal efficiencies and nitrite accumulation rates increased with higher COD/N ratios, indicating that high COD/N ratios can improve the competition between denitrifiers and nitrite-oxidizing bacteria for nitrite, leading to stable simultaneous nitritation and denitritation. The supplementation of SCSs further enhanced this high-efficiency N-removal process, with polybutylene succinate (PBS) and polycaprolactone (PCL) showing greater increases in N-removal via nitrite than poly-hydroxybutyrate (PHB) and poly-hydroxyalkanoate (PHA). Moreover, PBS showed the most significant increase in denitrification efficiency in anoxic conditions, while PHA was the most effective external SCS at a moderate level of dissolved oxygen. These findings suggest that the incorporation of external SCSs can facilitate the simultaneous nitritation and denitrification process in multi-cycle SBRs, underscoring the importance of selecting an appropriate SCS for optimizing nitrogen removal in sewage treatment projects.

Reuse of plastic waste as bio-carriers for domestic wastewater treatment – An approach towards circular economy

With the rapid growth of economic development, the circular economy has been raised in many internationalenvironmental and socio-economic conferences and forums nowadays. The recycling of plastic waste was oneof those aspects. This paper studied the potential of directly reusing plastic bottle caps as bio-carriers in movingbed bioreactors without further waste processing. As the caps had lower surface areas than the commercial carriers, total bacteria communities on the commercial carriers (K3) were 3-4 times higher than those on the bottle caps at different counting magnitudes. Nevertheless, the bottle caps could accommodate similar nitrogen-based bacteria (ammonia-oxidizing bacteria - AOB and nitrite-oxidizing bacteria - NOB density), which promotes well for the nitrification and denitrification processes. The COD and N-NH4 removal efficiencies were 92% and 86% on average, respectively. The high pollutant removal capacity reveals the very promising potential of bottle caps as bio-carriers for domestic wastewater treatment, promoting waste reduction and treatment expenses and presenting an excellent example of economic circular.

Performance of single PN/A reactor under wide fluctuation of nitrogen load.

Partial nitritation-anammox (PN/A) is a cost-effective technology in high ammonia-nitrogen wastewater treatment. However, PN/A is prone to instability as the ammonia-nitrogen sharply fluctuates. In this study, a packed bed reactor is employed to construct a single-stage PN/A system to investigate the operational characteristics and explore the denitrification mechanism. The effluent NH4+-N concentration, ammonia nitrogen removal rate (ARE), and total nitrogen removal rate (TNR) could be sustained at about 60mg/L, 80%, and over 70%, respectively, when the influent nitrogen load rate (NLR) is changed from 0.733 to 0.879kg-N/m3/day. Both ARE and TNR are decreased when NLR continues increasing to 1.026kg-N/m3/day. The influent NLR decreases from 0.879 to 0.147kg-N/m3/day, and ARE and TNR reached 98% and 85.4%, respectively. Therefore, the denitrification effect of the reactor could be recovered, and the excellent nitrogen removal capacity could be obtained within a wide range of influent NLR. Moreover, the high-throughput sequencing and metagenomic testing indicate that the Proteobacteria and Planctomycetes that the PN/A functional strains (i.e., ammonia-oxidizing bacteria (AOB) and anammox bacteria (AnAOB)) account for 38.8% in the sludge. The relative abundance of Nitrospira containing the nitrite-oxidizing bacteria (NOB) has dropped to 0.01%, and the functional gene nxr of the nitrite oxidation process is also inhibited. The relative expression of the functional gene is dominated by the short-range nitritation and anammox oxidation, which demonstrates that the nitrogen removal is mainly dominated by nitritation-anammox.

Microbial community dynamics in different floc size aggregates during nitrogen removal process upgrading in a full-scale landfill leachate treatment plant

Upgrading processes to reduce biodegradable organic substance addition is crucial for treating landfill leachate with high pollutant concentrations, aiding carbon emission reduction. Aggregate size in activated sludge processes impacts pollutant removal and sludge/water separation. This study investigated microbial community succession and driving mechanisms in different floc-size aggregates during nitrogen removal progress upgrade from conventional to partial nitrification–denitrification in a full-scale landfill leachate treatment plant (LLTP) using 16S rRNA gene sequencing. The upgrade and floc sizes significantly influenced microbial diversity and composition. After upgrading, ammonia-oxidizing bacteria were enriched while nitrite-oxidizing bacteria suppressed in small flocs with homogeneity and high mass transfer efficiency. Larger flocs enriched Defluviicoccus, Thauera, and Truepera, while smaller flocs enriched Nitrosomonas, suggesting their potential as biomarkers. Multi-network analyses revealed microbial interactions. A deep learning model with convolutional neural networks predicted nitrogen removal efficiency. These findings guide optimizing LLTP processes and understanding microbial community dynamics based on floc size.

Saturated dissolved oxygen-driven high-rate and ultrastable partial nitrification in municipal wastewater

Achieving stable and high-rate partial nitrification (PN) remains a worldwide technical conundrum in low-strength mainstream conditions. This study successfully achieved ultrarapid mainstream PN within 8 days under a saturated dissolved oxygen (DO) supply strategy, reaching a record-breaking PN rate of over 1.0 kg N m−3 d−1 treating municipal wastewater. Stable PN was maintained for over 200 days with an ultrahigh nitrite accumulation ratio of 98.5 ± 0.9 %, resilient to seasonal fluctuations in temperature (16.0–25.6 °C) and load (NH4+-N, 40–80 mg N/L). Kinetics revealed a remarkable 159.1-fold increase in the maximum activity ratio of ammonia-oxidizing bacteria (AOB) to nitrite-oxidizing bacteria (NOB). The faster response of AOB to saturated DO stimulated its highest activity difference with NOB, contributing to the AOB (Nitrosomonas oligotropha) boom and the elimination of NOB groups (−99.9 %). Our results highlight the importance of promoting AOB rather than solely focusing on NOB suppression for initiating and stabilizing high-rate mainstream PN.

Metabolite release by nitrifiers facilitates metabolic interactions in the ocean.

Microbial chemoautotroph-heterotroph interactions may play a pivotal role in the cycling of carbon in the deep ocean, reminiscent of phytoplankton-heterotroph associations in surface waters. Nitrifiers are the most abundant chemoautotrophs in the global ocean, yet very little is known about nitrifier metabolite production, release, and transfer to heterotrophic microbial communities. To elucidate which organic compounds are released by nitrifiers and potentially available to heterotrophs, we characterized the exo- and endometabolomes of the ammonia-oxidizing archaeon Nitrosopumilus adriaticus CCS1 and the nitrite-oxidizing bacterium Nitrospina gracilis Nb-211. Nitrifier endometabolome composition was not a good predictor of exometabolite availability, indicating that metabolites were predominately released by mechanisms other than cell death/lysis. Although both nitrifiers released labile organic compounds, N. adriaticus preferentially released amino acids, particularly glycine, suggesting that its cell membranes might be more permeable to small, hydrophobic amino acids. We further initiated co-culture systems between each nitrifier and a heterotrophic alphaproteobacterium, and compared exometabolite and transcript patterns of nitrifiers grown axenically to those in co-culture. In particular, B vitamins exhibited dynamic production and consumption patterns in nitrifier-heterotroph co-cultures. We observed an increased production of vitamin B2 and the vitamin B12 lower ligand dimethylbenzimidazole by N. adriaticus and N. gracilis, respectively. In contrast, the heterotroph likely produced vitamin B5 in co-culture with both nitrifiers and consumed the vitamin B7 precursor dethiobiotin when grown with N. gracilis. Our results indicate that B vitamins and their precursors could play a particularly important role in governing specific metabolic interactions between nitrifiers and heterotrophic microbes in the ocean.

Activation of iron oxides through organic matter-induced dissolved oxygen penetration depth dynamics enhances iron-cycling driven ammonium oxidation in microaerobic granular sludge

The iron redox cycle can enhance anammox in treating low-strength ammonia wastewater. However, maintaining an effective iron redox cycle and suppressing nitrite-oxidizing bacteria in a one-stage partial nitritation and anammox (PN/A) process poses challenges during long-term aeration. We proposed a novel and simple strategy to achieve an efficient iron redox cycle in an iron-mediated anoxic-microaerobic (A/O) process by controlling organic matter (OM) at medium-strength levels (30–110 mg COD/L) in microaerobic granular sludge (MGS)-dominated reactor. The developed A/O process consistently achieved >90 % OM removal and >75 % nitrogen removal. Medium-strength OM varied the penetration depths of dissolved oxygen (DO) in MGS, regulating redox conditions and promoting redox reactions across MGS layers, thus activating accumulated inert iron oxides. Ammonia-oxidizing bacteria (Nitrosomonas), iron-reducing bacteria (e.g., Ignavibacterium, Geobacter), and anammox bacteria (Ca. Kuenenia) coexisted harmoniously in MGS. This coexistence ensured high anammox and Feammox rates along with a robust iron redox cycle, thereby mitigating the adverse impacts of fluctuating DO and OM on one-stage PN/A process stability. The identification of iron reduction-associated genes within Ca. Kuenenia, Ignavibacterium, and Geobacter suggests their potential roles in supporting Feammox coupled in one-stage PN/A process. This study introduces an iron-cycle-driven A/O process as an energy-efficient alternative for simultaneous carbon and nitrogen removal from low-strength wastewater.

NOB suppression by organic pollutants in mainstream single-stage partial nitrification/anammox (PN/A) process: Experimental evidence and modelling investigation

Effectively and stably suppressing nitrite oxidizing bacteria (NOB) under mainstream conditions is difficult for the wide application of the partial nitrification/anammox (PN/A) process. In this work, a continuous-flow single-stage PN/A reactor experiencing nitrate accumulation was restored by increasing the influent chemical oxygen demand (COD) concentrations. Experimental results showed that the total nitrogen removal efficiency quickly increased from 70 ± 2.54 % to 84 ± 1.21 % while effluent nitrate concentrations dropped from 30 ± 4.07 mg N/L to 10 ± 3.18 mg N/L following increasing the influent COD concentrations. A modified and calibrated mathematical model was employed to quantify the nitrogen conversion process and microorganism distributions under various influent COD levels. The detailed nitrogen metabolism network was calculated by labeling the source and sink of nitrite and nitrate. Simulation results revealed that influent COD suppressed NOB activity and enhanced anammox bacteria (AMX) activity in a short-term dynamic state while leading to the elimination of NOB and enrichment of the AMX under long-term pseudo-steady state. N-metabolism simulation results indicated that more nitrite (from 46.62 mg N/L to 55.05 mg N/L) was diverted to AMX with the assistance of heterotrophic bacteria (HET). NOB was outcompeted due to the dual pressure of competing with HET and AMX for oxygen and nitrite, respectively. Microbial analysis solidly supported these findings, showing that Candidatus_kuenenia, Thauera and Denitratisoma were enriched, while the relative abundance of Nitrospira decreased after COD addition. These findings demonstrate the potential of using organic pollutants to suppress and eliminate NOB, thereby enhancing the stability and efficiency of mainstream PN/A reactors.

Free Ammonia Strategy for Nitrite-Oxidizing Bacteria (NOB) Suppression in Mainstream Nitritation Start-Up

The partial nitritation (PN)–anammox (PN/A) process offers a sustainable alternative to nitrogen management in wastewater treatment, addressing the high costs and increasing the low eco-friendliness associated with traditional nitrification/denitrification processes. Stable partial nitritation (PN) is critical for effective PN/A operation, and this study specifically focused on the need to suppress nitrite-oxidizing bacteria (NOB) to facilitate the enrichment of ammonia-oxidizing bacteria (AOB). Utilizing two sequencing batch reactors (SBRs), PN1 and PN2 with different free ammonia (FA) concentrations, this study aimed to evaluate the NOB suppression strategy while enriching AOB. The PN2 reactor, which operated with a higher initial FA concentration (50 mg/L), successfully maintained high nitritation activity, with 96.1% ammonium removal efficiency (ARE) and 95.1% nitrite accumulation efficiency (NAE) at reduced influent NH4+-N concentrations (50 mg NH4+-N/L, FA 10 mg/L). In contrast, PN1 showed inadequate NOB suppression due to lower FA concentrations (10 mg/L). These results suggest that initiating the nitritation process with higher FA concentrations can effectively suppress NOB, enhancing the stability and efficiency of PN/A processes in mainstream applications.

Long-term effect of phenol, quinoline, and pyridine on nitrite accumulation in the nitrification process: performance, microbial community, metagenomics and molecular docking analysis

Phenol, quinoline, and pyridine, commonly found in industrial wastewater, disrupt the nitrification process, leading to nitrite accumulation. This study explores the potential mechanisms through which these biotoxic organic compounds affect nitrite accumulation, using metagenomic and molecular docking analyses. Despite increasing concentrations of these compounds from 40 to 160 mg/L, ammonia nitrogen removal was not hindered, and stable nitrite accumulation rates exceeding 90 % were maintained. Additionally, these compounds inhibited nitrite-oxidizing bacteria (NOB) and enriched ammonia-oxidizing bacteria (AOB) in situ. As the concentration of these compounds rose, protein (PN) and polysaccharide (PS) concentrations also increased, along with a higher PN/PS ratio. Metagenomic analysis further revealed an increase in hao relative abundance, while microbial community analysis showed increased Nitrosomonas abundance, which contributed to nitrite accumulation stability. Molecular docking indicated that these compounds have lower binding energy with hydroxylamine oxidoreductase (HAO) and nitrate reductase (NAR), theoretically supporting the observed sustained nitrite accumulation.

Impact of N loading on Microbial Community Structure and Nitrogen Removal of an Activated Sludge Process with Long SRT for municipal wastewater treatment

Biological nutrient removal process is pivotal in controlling nitrogen and phosphorus from municipal wastewater treatment and reclamation plants in the water environment. The efficiency of municipal wastewater treatment is affected by the fluctuation of influent quality. In this study, the impact of N loading on the removal efficiency and the microbial community structure of activated sludge with long sludge retention time were investigated. The results showed that the effluent NH4+-N and NO2--N concentration were lower than 0.5 mg/L, and the TN removal efficiencies were 79.77% and 65.80%, respectively. The dehydrogenase activity, specific oxygen uptake rate, specific nitrification rate, and specific denitrification rate increased as the N loading increasing. At the phylum level, the most dominant bacterial was affiliated with Proteobacteria in the two systems, accounting for 60.65% and 48.62%, respectively. At the genus level, Acinetobacter, Thauera, and Zoogloea played significant roles in denitrification. The ammonia oxidation bacteria and nitrite-oxidizing bacteria were of the genus Nitrosomonas (r-strategist) and Nitrospira (K-strategist).

Synergy between Nitrogen Removal and Fermentation Bacteria Ensured Efficient Nitrogen Removal of a Mainstream Anammox System at Low Temperatures.

Simultaneous partial nitrification, anammox, denitrification, and fermentation (SNADF) is a novel process achieving simultaneous advanced sludge reduction and nitrogen removal. The influence of low temperatures on the SNADF reactor was explored to facilitate the application of mainstream anammox. When temperature decreased from 32 to 16 °C, efficient nitrogen removal was achieved, with a nitrogen removal efficiency of 81.9-94.9%. Microbial community structure analysis indicated that the abundance of Candidatus Brocadia (dominant anaerobic ammonia oxidizing bacteria (AnAOB) in the system) increased from 0.03% to 0.18%. The abundances of Nitrospira and Nitrosomonas increased from 1.6% and 0.16% to 2.5% and 1.63%, respectively, resulting in an increase in the ammonia-oxidizing bacteria (AOB) to nitrite-oxidizing bacteria (NOB) abundance ratio from 0.1 to 0.64. This ensured sufficient nitrite for AnAOB, promoting nitrogen removal. In addition, Candidatus Competibacter, which plays a role in partial denitrification, was the dominant denitrification bacteria (DNB) and provided more nitrite for AnAOB, facilitating AnAOB enrichment. Based on the findings from microbial correlation network analysis, Nitrosomonas (AOB), Thauera, and Haliangium (DNB), and A4b and Saprospiraceae (fermentation bacteria), were center nodes in the networks and therefore essential for the stability of the SNADF system. Moreover, fermentation bacteria, DNB, and AOB had close connections in substrate cooperation and resistance to adverse environments; therefore, they also played important roles in maintaining stable nitrogen removal at low temperatures. This study provided new suggestions for mainstream anammox application.

Achieving nitrite shunt using in-situ free ammonia enriched by natural zeolite: Pilot-scale mainstream anammox with flexible nitritation strategy

The mainstream partial nitritation/anammox (PN/A) process represents a significant innovation in decarbonizing municipal wastewater treatment. However, its implementation is considerably hampered by the challenge of stable nitrite supply. In this study, a pilot-scale PN/A system receiving real sewage (20 m3) was operated at room temperature for nearly one year. Remarkable PN performance with relatively high nitrite accumulation ratio of 75.04 ± 10.05 % was obtained via in-situ free ammonia (FA) strategy. The ammonium concentration enriched in the zeolite increased significantly by 548.8 times compared to that in the aqueous phase by ion exchange. This substantial increase robustly inhibited nitrite-oxidizing bacteria (NOB), resulting in high relative abundance ratio of ammonia-oxidizing bacteria (AOB) to NOB of 37.93 ± 12.61 in the zeolite biofilm, compared to 10.22 ± 1.67 in suspended floc sludge. The significant differences in FA concentrations between zeolite biofilm and suspended floc sludge resulted in distinct spatial distribution disparities of AOB and NOB, which were central to achieving stable nitrite accumulation without complex multiple selective pressures. Consequently, compliant effluent with total nitrogen of 10.91 ± 4.23 mg N/L was achieved at 10.4-31.1 °C without external carbon source addition. The biocarriers in the anammox process played a key role in enhancing functional genes and electron flow, supporting anammox-dominated nitrogen removal. This study presents a flexible and adaptable strategy for mainstream nitrite shunting, highlighting its potential for large-scale implementation of mainstream anammox treatment.