Nitrogen Removal Pathway Research Articles

High rates of nitrogen removal in aerated VFCWs treating sewage through C-N-S cycle

The efficiency of deep aerated vertical flow constructed wetlands (DA-VFCWs) being operated in Hyderabad, India, was evaluated herein using physicochemical analysis and 16S rRNA amplicon sequencing. The results showed 2–4-fold higher removal rate coefficients for Biochemical oxygen demand (1.32–––3.53 m/d) and nitrogen (0.88––1.36 m/d) in DA-VFCWs than those of passive VFCWs. Elevated sulfate concentration in the DA-VFCWs effluent (84–113 mg/L) indicated possibility of sulfur-driven autotrophic denitrification (SDAD) as a major pathway operating in these wetlands besides the classical nitrogen removal pathways. The presence of nitrifiers (3.09–10.02 %), heterotrophic and aerobic denitrifiers (0.79–0.83 %), anammox bacteria (1.31–2.22 %) and SDAD bacteria (0.08–0.73 %) in the biofilm samples collected from the DA-VFCWs exemplify an interplay of Carbon-Nitrogen-Sulfur cycles in these systems. If proven, the presence of an operational SDAD pathway in DA-VFCWs can help reduce surface area requirement in VFCWs substantially besides alleviating biological clogging of the wetland substrate.

Synchronous in-situ sludge reduction and enhanced denitrification through improving electron transfer during endogenous metabolisms with Fe(Ⅱ) addition

The creation of large amounts of excess sludge and residual nitrogen are critical issues in wastewater biotreatment. This study introduced Fe(II) into an oligotrophic anaerobic reactor (OARFe) that was implemented to modify an anoxic-oxic process to motivate in-situ sludge reduction and enhance denitrification under an effective electron shuttle among organic matter, nitrogen, and Fe. The addition of 15 mg L−1 Fe(II) resulted in a sludge reduction efficiency reached 32.0% with a decreased effluent nitrate concentration of 33.3%. This was mostly attributed to the electron transfer from Fe(II) to organic matters and nitrogen species in OARFe. The participation of Fe(II) led to the upregulation of Geothrix and Terrimonas, which caused active organic matter hydrolysis and cell lysis to stimulate the release of extracellular polymeric substances (EPS) and substance transfer between each layer of EPS. The higher utilization of released bioavailable dissolved organic matter improved endogenous denitrification, which can be combined with iron autotrophic denitrification to realize multiple electron donor-based nitrogen removal pathways, resulting in an increased nitrate removal rate of 58.2% in the absence of external carbon sources. These functional bacteria associated with the transformation of nitrogen and carbon and cycling between ferrous and ferric ions were enriched in OARFe, which contributed to efficient electron transport occurred both inside and outside the cell and increased 2,3,5-triphenyltetrazolium chloride electronic transport system activity by 46.9%. This contributed to the potential operational costs of chemical addition and sludge disposal of Fe-AO being 1.9 times lower than those of conventional A2O processes. These results imply that the addition of ferrous ions to an oligotrophic anaerobic zone for wastewater treatment has the potential for low-cost pollution control.

Biological nitrogen removal characteristics and mechanisms of a novel salt-tolerant strain Vibrio sp. LV-Q1 and its application capacity for high-salinity nitrogen-containing wastewater treatment

High concentration of salts in wastewater can adversely affects the metabolic activities and growth of microorganisms, resulting in reduced efficiency of biological nitrogen removal. Screening high-efficiency heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria with salt tolerance is important for the wastewater treatment. In this study, a novel salt-tolerant strain LV-Q1 capable of HN-AD was isolated and identified as Vibrio sp. This strain demonstrates a salinity tolerance range of 0–8 %. Notably, strain LV-Q1 exhibited exceptional nitrogen removal efficiencies (at initial concentrations of 100 mg/L), achieving 97.97 % for NH4+-N, 100 % for NO3--N, and approximately 100 % for mixed nitrogen (NH4+-N and NO3--N) at a salinity of 5 %. Optimal ammonium removal parameters for LV-Q1 were established. The nitrogen removal pathway of LV-Q1 primarily involves direct assimilation of ammonium and its conversion into N2 through HN-AD. Analysis of the salt tolerance mechanism revealed LV-Q1's ability to equilibrate osmotic pressure through the accumulation of amino acids and betaine, production of antioxidative enzymes or extracellular protein, and utilization of its quorum sensing (QS) properties to withstand high-salinity environments. The strain LV-Q1 exhibited strong resistance to salinity shock in bioaugmentation within a moving bed biofilm reactor (MBBR), markedly enhancing nitrogen and carbon removal performance with an improved removal efficiency of 26.74 % and 10.15 % averagely at the end of the reaction at a salinity of 5 %. Moreover, Vibrio sp. LV-Q1 successfully proliferated and dominated, and its relative abundance gradually increased to 59.81 %. This study contributes new insights and resources for the treatment of nitrogen in saline effluents.

Genomics and metabolic characteristics of simultaneous heterotrophic nitrification aerobic denitrification and aerobic phosphorus removal by Acinetobacter indicus CZH-5

This study provides for the first time a systematic understanding of Acinetobacter indicus CZH-5 performance, metabolic pathway and genomic characteristics for aerobic nitrogen (N) and phosphorus (P) removal. Acinetobacter indicus CZH-5 showed promising performance in heterotrophic nitrification aerobic denitrification and aerobic phosphorus removal. Under optimal conditions, the maximum ammonia-N, total nitrogen and orthophosphate-P removal efficiencies were 90.17%, 86.33%, and 99.89%, respectively. The wide tolerance range suggests the strong environmental adaptability of the bacteria. The complete genome of this strain was reconstructed. Whole genome annotation was used to re-construct the N and P metabolic pathways, and related intracellular substance metabolic pathways were proposed. The transcription levels of related functional genes and enzyme activities further confirmed these metabolic mechanisms. N removal was achieved via the nitrification–denitrification pathway. Furthermore, CZH-5 exhibited significant aerobic P uptake, with phosphate diesters as the main species of intracellular P.

Recurrent multi-stressor floc treatments with sulphide and free ammonia enabled mainstream partial nitritation/anammox

Selective suppression of nitrite-oxidising bacteria (NOB) over aerobic and anoxic ammonium-oxidising bacteria (AerAOB and AnAOB) remains a major challenge for mainstream partial nitritation/anammox implementation, a resource-efficient nitrogen removal pathway. A unique multi-stressor floc treatment was therefore designed and validated for the first time under lab-scale conditions while staying true to full-scale design principles. Two hybrid (suspended + biofilm growth) reactors were operated continuously at 20.2 ± 0.6 °C. Recurrent multi-stressor floc treatments were applied, consisting of a sulphide-spiked deoxygenated starvation followed by a free ammonia shock. A good microbial activity balance with high AnAOB (71 ± 21 mg N L−1 d−1) and low NOB (4 ± 17 % of AerAOB) activity was achieved by combining multiple operational strategies: recurrent multi-stressor floc treatments, hybrid sludge (flocs & biofilm), short floc age control, intermittent aeration, and residual ammonium control. The multi-stressor treatment was shown to be the most important control tool and should be continuously applied to maintain this balance. Excessive NOB growth on the biofilm was avoided despite only treating the flocs to safeguard the AnAOB activity on the biofilm. Additionally, no signs of NOB adaptation were observed over 142 days. Elevated effluent ammonium concentrations (25 ± 6 mg N L−1) limited the TN removal efficiency to 39 ± 9 %, complicating a future full-scale implementation. Operating at higher sludge concentrations or reducing the volumetric loading rate could overcome this issue. The obtained results ease the implementation of mainstream PN/A by providing and additional control tool to steer the microbial activity with the multi-stressor treatment, thus advancing the concept of energy neutrality in sewage treatment plants.

Tapping the potential of anammox upgrading in full-scale wastewater treatment plants through biofilm-enhanced anammox and denitrification

Implementing the Anammox process at a large scale remains a significant challenge. However, this study presents a potential approach for upgrading full-scale wastewater treatment plants (WWTPs) to incorporate the Anammox process in treating ammonium-rich wastewater. The research focuses on a WWTP at a duck breeding farm that utilizes an anaerobic/microaerobic/anoxic/oxic (A/M/A/O) process. With 83% removal of total nitrogen, the contribution of anammox and denitrification nitrogen removal in the entire process was 52% and 31%, respectively. The study tracks the changes in water quality and analyzes the potential of simultaneous Anammox-denitrification processes to mitigate nitrate accumulation. Porous carriers were introduced into the primary anaerobic tank (Anaerobic T1) and the primary microaerobic tank (Microaerobic T1) and were harvested on the 90th day. The results revealed that the specific Anammox activity (SAA) of the microaerobic biofilm reached 252.06 mg N/g VSS·d, which was 6.0 times and 3.7 times higher than that of suspended sludge and anaerobic biofilm, respectively. The microaerobic biofilm demonstrated the capacity for simultaneous Anammox-denitrification, with anammox and denitrification rates of 65.35 and 36.3 nmol N/g·h, respectively. These rates were significantly higher than those observed in the anaerobic biofilm and suspended sludge. Furthermore, the main partial denitrification bacteria identified were Ignavibacterales UTCHB1 (57.75%) and Trichococcus (5.12%), which contributed nitrite to Candidatus Brocadia (1.54%). The findings highlight the importance of creating a suitable microenvironment using porous carriers, which facilitates the Anammox pathway for nitrogen removal in biofilms. This approach presents a potential retrofitting and upgrading strategy for existing WWTPs.

Achieving simultaneous removal of carbon and nitrogen by an integrated process of anaerobic membrane bioreactor and flow-through biofilm reactor

In this study, a combined system consisting of an anaerobic membrane bioreactor (AnMBR) and flow-through biofilm reactor/CANON (FTBR/CANON) was developed to simultaneously remove carbon and nitrogen from synthetic livestock wastewater. The average removal efficiencies of total nitrogen (TN) were 64.2 and 76.4% with influent ammonium (NH4+-N) concentrations of approximately 200 and 500 mg/L, respectively. The COD removal efficiencies were higher than 98.0% during the entire operation. Mass balance analysis showed that COD and TN were mainly removed by the AnMBR and FTBR/CANON, respectively. The anammox process was the main nitrogen removal pathway in the combined system, with a contribution of over 80%. High functional bacterial activity was observed in the combined system. Particularly, an increase in the NH4+-N concentration considerably improved the anammox activity of the biofilm in the FTBR/CANON. 16S rRNA high-throughput sequencing revealed that Methanosaeta, Candidatus Methanofastidiosum, and Methanobacterium were the dominant methanogens in the AnMBR granular sludge. In the CANON biofilm, Nitrosomonas and Candidatus Kuenenia were identified as aerobic and anaerobic ammonium-oxidizing bacteria, respectively. In summary, this study proposes a combined AnMBR and FTBR/CANON process targeting COD and nitrogen removal, and provides a potential alternative for treating high-strength wastewater.

Exploring organic matter conversion pathway and its effect on nitrogen removal in tidal flow constructed wetlands

Achieving effective nitrogen removal remains a significant challenge faced by constructed wetlands. Although organic matter is a crucial factor influencing nitrogen removal, little attention has been paid to the impact of organic matter conversion pathways on nitrogen removal in constructed wetlands. Here, we showed that endogenous microorganisms performing carbon internalization could be easily enriched in tidal flow constructed wetlands (TFCWs) under its special rhythmic cycle of anaerobic/aerobic operational mode. Endogenous microorganisms could translate influent carbon sources into intracellular carbons during the anaerobic stage and supply the carbon source for endogenous denitrification after the aerobic stage (rest period). Based on these findings, an innovative combined TFCW and Nitrifying-CW system was developed, and robust total nitrogen (TN) removal (82% on average) was achieved even under carbon source limiting conditions. This performance was a substantial improvement compared to the conventional single bed TFCW with multiple “tides” (corresponding to the multiple contact/rest periods) with TN removal of only 54% on average. Simultaneous nitrification-endogenous denitrification (SNED) was found to be the major nitrogen removal pathway in the proposed system. Compared with classical nitrification-denitrification, simultaneous nitrification-endogenous denitrification brings high nitrogen conversion rates and significantly reduces the demand for oxygen and organic carbon. Furthermore, microbial community analysis indicated that endogenous microorganisms such as Candidatus_Competibacter and Defluviicoccus were successfully enriched, accounting for 50.73% and 3.46% in CW1, and 25.25% and 1.76% in CW2, respectively. Together, these mechanisms allow the proposed system to achieve efficient TN removal.

Nutrients removal by high-rate activated sludge and its effects on the mainstream wastewater treatment

Conventional wastewater treatment technologies often involve energy-intensive processes, creating a need for more sustainable and efficient solutions. The high-rate activated sludge (HRAS) process, characterized by short sludge and hydraulic retention times, demonstrates proficient chemical oxygen demand (COD) removal with minimal oxidation and energy expenditure, positioning it as a promising approach for achieving energy-efficient wastewater treatment.This study addresses the nutrient removal efficiency of a demonstration-scale pilot plant (35 m3·d-1) operating for 497 days. It investigates the correlation between nutrient removal efficiencies, COD fractions, and suspended solids removal in real wastewater, without primary clarification.The process consistently achieves average removal efficiencies: 23% for total Kjeldahl Nitrogen (TKN), 7.3% for N-NH4+, 56% for total phosphorus (TP), and 11% for P-PO43-. The superior TP removal is attributed to the high particulate influent fractions (81%), underscoring the pivotal role of settling efficiency. The HRAS process effectively controls TP peak values, although its influence on TKN peaks is somewhat limited. This underscores HRAS's proficiency in mitigating phosphorus spikes in influent wastewater. In contrast, the lower TKN removal is associated with diminished particulate inlet fractions (46%).Nutrient removal mechanisms in HRAS mirror those of COD, primarily revolving around adsorption and entrapment processes, as opposed to conventional biological pathways. It also highlights the independence of oxidized COD, nitrogen, and phosphorus removal. Furthermore, a comprehensive analysis of the COD/TKN within HRAS effluent unveils the presence of a short-cut nitrification/denitrification process, supplemented by Anammox in the sidestream, as the most suitable pathway for downstream nitrogen removal. This revelation signifies a substantial advancement and positions HRAS as a potential initial stage of a partial nitritation/Anammox (PN/A) process within mainstream wastewater treatment.

How the subsurface wastewater infiltration systems with iron‑carbon micro-electrolytic substrate achieve high pollutants removal: A metabolomic view

Being an ecological engineering water treatment technology, subsurface wastewater infiltration system (SWIS) combines ecological principles with anaerobic and aerobic processes. Iron‑carbon micro-electrolysis (ICME) is a rapidly developing SWIS substrate type in recent years. In this study, microbial metabolomics was introduced into SWIS aiming to reveal the “black box” of enhanced nitrogen removal in SWIS assisted by ICME (namely ICMESWIS). Ultra performance liquid chromatography-mass spectrometry (UPLC-MS), principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and univariate analysis were carried out to classify the samples from SWIS and ICMESWIS. The results showed that compared with SWIS, 28.5 ± 2.6 %, 33.7 ± 1.3 % and 19.3 ± 0.4 % improvements were found for NO3−-N, NH4+-N and NO2−-N removals in ICMESWIS. According to the model's variable importance factor and fold change >2, metabolites with large differences were screened out. Metabolites, such as 1-(4-nitrophenyl)piperidine (ESI-) and 2-amino-1,3,4-octadecanetriol (ESI-) presented higher abundance in ICMESWIS, in consistent with the higher degradation of pollutants. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that nitrification, nitrate reduction, dissimilatory nitrate reduction, denitrification, anaerobic ammonia oxidation (anammox) are the main nitrogen removal pathways in ICMESWIS.

Microalgae biofilm photobioreactor and its combined process for long-term stable treatment of high-saline wastewater achieved high pollutant removal efficiency

High-saline wastewater treatment processes have a high energy demand, while microalgae-driven wastewater biotechnology is both economical and environmentally friendly, and the use of microalgae to treat high-saline wastewater can effectively reduce energy consumption. However, it is unclear whether the performance of microalgae reactor can be stable for a long time. In this study, an open continuous-flow biofilm photobioreactor using Dunaliella salina was innovatively constructed, and operated in combination with a membrane bioreactor (MBR) for high-saline wastewater treatment. The purpose of this study was to investigate the long-term performance of MBR, MBPBR alone and their combined operation for the removal of organic matter, phosphorus and nitrogen, explore the pathway and mechanism of nitrogen and phosphorus removal by MBPBR, and evaluate the feasibility of MBPBR-MBR integrated system for the treatment of saline wastewater. The MBR effectively removed most of the organic matter and converted ammonia nitrogen into nitrate nitrogen, which is a preferred nitrogen source for the growth of Dunaliella salina, thereby providing favorable water quality for Dunaliella salina in the subsequent MBPBR. The MBPBR, with a hydraulic retention time (HRT) of 72 h, achieved a NO3--N removal load of 74.9 g N/(m3∙d) and a removal efficiency of approximately 60 %, and the PO43--P removal load of 8.33 g P/(m3∙d) and removal efficiency of over 80 %. The MBR-MBPBR system demonstrated long-term stability, achieving removal efficiencies of 84.3 %, 81.6 %, and 91.0 % for COD, TN and PO43--P, respectively. Within the MBPBR, the growth of algae and their assimilation contributed 44.9 % and 38.7 % of the nitrogen and phosphorus removal, respectively. The abundance of Dunaliella salina in the MBPBR remained at approximately 90 % for over 40 days of operation, providing an opportunity for Dunaliella salina recovery. These findings provide valuable insights into the utilization of microalgae for the treatment of high-salinity wastewater, while also supporting the practical implementation of microalgae-based treatment methods for such wastewater.

Approach enhancing nitrate removal from actual municipal wastewater by integrating electric-magnetic field with Fe0 in UMSR reactor

This study aimed to investigate the effectiveness of combining zero-valent iron (Fe0) with an electric-magnetic field (EMF) with an intensity of 48-mT generated by an alternating current (AC) of 6 V and 50 Hz applied deep within a reactor using actual wastewater obtained from a secondary stage of a wastewater treatment plant (WWTP). The lab-scale experiment was conducted using the UMSR reactor, maintaining a controlled temperature of 25 ± 2 °C and a hydraulic retention time (HRT) of 5-hour, with a dissolved oxygen (DO) level below 1.5 mg/L. Fine Fe0 particles with a size of ≤ 0.3 µm were added into the reactor at a concentration of 3 g/L. The actual wastewater had an approximate nitrate nitrogen (NO3–-N) concentration of 30 ± 2 mg/L, and the substrate mass ratio of carbon to nitrogen (C:N) was at 1:1. The results examined the performance of the EMF-Fe0 system in terms of nitrate removal efficiency and the enrichment of denitrifying bacteria, achieving a nitrate removal rate of 67 % with an effluent NO3–-N concentration of 10 mg/L and a chemical oxygen demand (COD) concentration of 11.8 mg/L at removal rate of 60 %. Illumina sequencing revealed various enriched denitrifying bacteria after coupling EMF with Fe0, including autotrophic Nitrospira (NOB) at 4 %, denitrification bacteria like Hydrogenophaga at 1.3 %, Hyphomicrobium at 1 %, and Sphingobium at 5 %, along with thermophilic bacteria Thermus at 1.1 % and Meiothermus at 3.5 %. PICRUSt predicted bacterial functions and analyzed interactions, while KEGG and GO analyses linked nitrogen removal pathways to specific species, genes, and enzymes in the EMF-Fe0 system.

Partial denitrification-anammox granular sludge domesticated from high-strength anammox granules and the high-efficiency performance in treating low-nitrogen wastewater

Partial denitrification (PD) is an effective approach to supply nitrite for the anaerobic ammonium oxidation (anammox) reaction, and the PD-anammox process can achieve nearly 100 % nitrogen removal when denitratation, denitritation and anammox processes are well balanced. However, most of the previous studies initiated the PD-anammox process by a mixed inoculation of denitrification and anammox sludges, leading to low anammox bacteria abundance and a risk of unstable operation in the long-term. To address this, this study used high-strength anammox granules as seed sludge for rapid PD-anammox process start-up. During the continuous operation, nitrogen removal efficiencies of 97.6 ± 1.6 % and 91.0 ± 2.0 % were attained under influent conditions of 159.4 ± 3.4 mg N/L and 78.1 ± 3.5 mg N/L, respectively. In addition, the nitrite conversion rate of the sludge reached 5.34 ± 0.37 g N/g VSS/d during the operation, and the anammox activity was stable at 0.27–0.41 g N/g VSS/d, indicating a synergistic PD-anammox relationship. Furthermore, PD bacteria were efficiently enriched in the reactor within a short period of time, with Thauera accounting for 7.6 % of the relative abundance. Notably, the abundances of dominating anammox bacteria, Ca. Brocadia and Ca. Kuenenia were 16.6 % and 4.9 %, respectively, which were among the highest of the reported PD-anammox processes. High anammox microbial abundance is conducive to ensuring the dominance and stability of the anammox reaction, enabling efficient nitrite conversion and nitrogen removal throughout the continuous operation. Finally, nitrogen removal pathways based on mass balance were proposed to clarify the fate of different nitrogen compounds in the PD-anammox system.

Optimization of nitrogen removal conditions based on response surface methodology and nitrogen removal pathway of Paracoccus sp. QD-19

The strain Paracoccus sp. QD-19 was isolated from the sludge-water mixture of aerobic tanks at the southern wastewater treatment plant in Shenyang, China. The optimal nitrogen removal conditions for strain QD-19 were determined using the Plackett–Burman design, path of steepest ascent method, and response surface methodology (RSM). The optimum nitrogen removal conditions were C/N 12.93, temperature 37 °C, and shaking speed 175.50 r/min. Strain QD-19 achieved 83.82 ± 0.80 % nitrogen removal efficiency at 10 h under optimum conditions. Functional enzyme-encodinge genes amplified via 16S rRNA sequence analysis included amoA, hao, napA, nirS, nirK, norB, and nosZ. The results revealed that NH4+-N → NH2OH → NO2−-N → NO3‐-N → NO2−-N → NO → N2O → N2 was the pathway for heterotrophic nitrification - aerobic denitrification. The strain was used to treat wastewater from a sewage treatment plant under optimal response surface methodology conditions. As a result, the TN removal efficiency was 77.11 %. The findings demonstrated that strain QD-19 exhibits favorable potential for heterotrophic nitrification and aerobic denitrification (HN-AD) of actual wastewater, presenting a promising application for biological wastewater treatment.

Stimulating extracellular polymeric substances production in integrated fixed-film activated sludge reactor for advanced nitrogen removal from mature landfill leachate via one-stage double anammox

Introducing carbon sources to achieve nitrogen removal from mature landfill leachate not only increases the costs and carbon emissions but also inhibits the activity of autotrophic bacteria. Thus, this study constructed a double anammox system that combines partial nitrification-anammox (PNA) and endogenous partial denitrification-anammox (EPDA) within an integrated fixed-film activated sludge (IFAS) reactor. In this system, PNA primarily contributes to nitrogen removal pathways, achieving a nitrite accumulation rate of 98.23%. The production of extracellular polymer substances (EPS) in the IFAS reactor is stimulated by introducing co-fermentation liquid. Through the utilization of EPS, the system effectively achieves EPDA with the nitrite transformation rate of 97.20%. Under the intermittent aeration operation strategy, EPDA combined with PNA and anammox in the oxic and anoxic stages enhanced the nitrogen removal efficiency of the system to 99.70 ± 0.12%. The functional genus Candidatus kuenenia became enriched in biofilm sludge, while Thauera and Nitrosomonas predominated in floc sludge.

Microbial stratification protects denitrifying anaerobic methane oxidation archaea and bacteria from external oxygen shock in membrane biofilm reactor

Different gradients of dissolved oxygen (DO) regulate the microbial community and nitrogen removal pathways of denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (Anammox) coupled process in a batch biofilm reactor. Under completely anaerobic condition, approximately 72 mg NO3–-N/L was removed at a daily rate of 6.55 mg N/L, whereas a peak accumulation of 95 mg NO3–-N/L was observed during DO reached 0.5 mg/L. There is a decrease in the abundance of Candidatus Methylomirabilis (24.1%), Candidatus Methanoperedens (23.3%), and Candidatus Kuenenia (22.6%) to below 5% when DO levels reached 0.2 mg/L. Moreover, key genes associated with the reverse methanogenesis (mcrA) and anaerobic ammonium oxidase (hzo) decreased. These findings indicate that during oxygen shock, methanotrophs and denitrifiers replace Anammox bacteria on the outer sphere of the biofilm, whereas DAMO bacteria and archaea are protected from external oxygen shock due to the microbial stratification of biofilm.

A pilot study of situ sludge fermentation-driven multiple biological nitrogen removal pathways (SFBNR): Revealing microbial synergy mechanism based on co-occurrence network analysis

The sludge fermentation-driven biological nitrogen removal (SFBNR) has garnered increasing attention due to its efficient carbon resource utilization from waste activated sludge (WAS). This study successfully extended the application of this technique to a 38 m3 reactor, facilitating a daily ultra-low carbon to nitrogen ratio (<1) wastewater treatment capacity of 16 tons and a WAS capacity of 500 L. After 185-days operation, the system demonstrated commendable performance with a denitrification efficiency (DNE) of 93.22 % and a sludge reduction efficiency (SRE) of 72.07 %. To better understand the potential mechanisms, various functional bacteria interactions were revealed by co-occurrence network analysis. The results unveiled module hubs (e.g., Anaerolineaceae, Denitratisoma, and Candidatus Brocadia) and connectors (e.g., Tuaera and Candidatus Alysiosphaera) in the network exhibited synergistic relationships facilitated by carbon metabolism and nitrogen cycling. Furthermore, the interaction between biofilm sludge (BS) and suspended sludge (SS) contributed to the in-situ enrichment of anaerobic ammonium oxidizing bacteria (AnAOB), whose abundance in BS reached 1.8 % (200-times higher than in SS) after six months, and the suspend-biofilm interface served as a hotspot for anammox activity.

Algal-bacterial granular sludge for simultaneous denitrification and yttrium removal

This study investigated the performance and stability of algal-bacterial granular sludge (ABGS) for the treatment of inorganic wastewater containing ammonia and Y(III), which aimed to shed light on efficient treatment of yttrium-rich rare earth mine wastewater. By seeding heterotrophic granules and dosing exogenous algae, cationic polyacrylamide and sodium acetate in a photo-sequencing batch reactor, the algal and granular symbiosis system was successfully established on Day 50, and mature ABGS was harvested on Day 165. When the concentration of Y(III) was no more than 4 mg/L, the system showed good performance for simultaneous removal of nitrogen and Y(III). However, further increasing the Y(III) concentration to 30 mg/L in Days 144–210 gradually inhibited the bacterial activity and killed most of the functional algae, resulting in significant decrease of the activity of bacteria and chlorophyll a content after Days 135 and 180, as well as the oxygen yield rate of the algae and ammonia oxidation rate after Days 140 and 95. The removal efficiency of total inorganic nitrogen maintained mostly above 90% in Days 46–153, and gradually decreased to approximately 50% in Days 154–210. The Y(III) removal efficiency remained above 98% in Days 96–125, and gradually decreased to approximately 60% in Days 126–210. Most of the algal genera disappeared on Day 210. Conversely, the increase of the relative abundance of Nitrosomonas and Thauera was observed. The nitrogen removal pathways included denitrification and algal assimilation. However, the contribution rate of algae gradually decreased from 22% to 4% due to the accumulation of Y.

Spatial difference in nitrogen removal pathways and microbial functional diversity in an EGSB reactor during the start-up of PD/Anammox

The start-up of a relatively high nitrogen load PD/Anammox in an EGSB reactor was achieved through strategies of bioaugmentation, mass transfer enhancement, and COD/NO3−-N control, with NRR of 5.2 g N/L/d. Longitudinal heterogeneity in EGSB reactor induced divergent nitrogen conversion pathways and enriched different functional microorganisms between stratified sludge. Along the elevation of the reactor, the proportion of removed nitrogen through anammox increased continuously from bottom, middle and up, which were 65.0 %, 79.8 %, and 84.1 %, respectively, consistent with the trend of ex-situ activities calculated with Gompertz model. The bottom zone played a role in mixed nitrogen conversion to provide NO2−-N accumulation and nitrogen removal, with higher abundance of Thauera, Denitratisoma and Ignavibacterium. The middle part was enriched Candidatus_Kuenenia (12.51 %), and up inhibited completed denitrification, together forming the anammox dominant zone. The proposed functional zones in the EGSB reactor provided approaches for the optimisation of high-load PD/Anammox systems.

Enhanced nitrogen removal driven by S/Fe2+ cycle in a novel hybrid constructed wetland

Insufficient carbon sources and oxygen limit the nitrogen removal efficiency of constructed wetlands. In this study, a novel hybrid constructed wetland (SS-HCW) was designed by combining sulfur-siderite filler and drop aeration to enhance nitrogen removal. The results showed that the TN removal rate increased by 37.74%. Further analysis of nitrogen removal pathways found that sulfur autotrophic denitrification was dominant. In addition, SO42− and Fe2+ generated during nitrogen removal can serve as trace elements to increase the growth rate and chlorophyll content of wetland plants by 50.0% and 34.2%, respectively. The microbial analysis results showed that the sulfur-siderite filler changed community structure, with Thiobacillus and Sulfurimonas as the dominant genera, and enriched some bacteria with sulfur and iron oxidation capabilities such as Thiomonas and Ferritrophicum.