Autotrophic Denitrification Process 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.

Phosphorus removal performance of Sulfide-Based autotrophic denitrification process

Sulfide-based Autotrophic Denitrification (SAD) Process can simultaneous remove sulfurous and nitrogenous pollutants, showing a promising prospect. Beyond our expectation, it also has an efficient phosphorus (P) removal performance. The operation results demonstrated that when the influent concentration of nitrate, sulfide and phosphate was 80.55 ± 2.98 mg N/L, 380.15 ± 20.83 mg S/L and 47.70 ± 4.35 mg P/L respectively, their removal efficiency was 87.63 ± 3.12 %, 99.61 ± 1.02 % and 85.38 ± 4.07 % at the HRT of 8.8 h. However, the phosphorus removal performance disappeared once the effluent pH dropped below 8.0. The random forest regression model revealed that effluent pH had the most significant impact on phosphorus removal performance. This finding was corroborated by a mathematical model that related the phosphate removal load to effluent pHs. The combination of batch test, Standard Measurements and Testing (SMT) and X-Ray Diffraction (XRD) method revealed that the phosphorus removal performance of SAD process came from the synergies of biological phosphorus removal (29.71 ± 3.21 %) and bio-induced phosphate precipitation (70.29 ± 3.21 %). The analysis of key functional genes coding for nitrogen, sulfur and phosphorus metabolism offered an explanation for their metabolic pathways especially the phosphorus removal pathway. The study would provide some new information for the development of SAD process.

Understanding nitrogen removal and N2O emission mechanisms in an anaerobic-swing-anoxic-oxic (ASAO) continuous plug-flow system for low C/N municipal wastewater

This study aims to investigate effects of dissolved oxygen (DO) levels and aerated hydrodynamic retention time (HRT) on nitrogen removal and nitrous oxide (N2O) emissions in a novel anaerobic-swing-anoxic-oxic (ASAO) continuous plug-flow system for treating low carbon to nitrogen ratio municipal wastewater. The swing zones had varying DO levels and volumes, deciding the aerated HRT of the ASAO system. Results showed that low DO level (0.8–1.0 mg/L) and short aerated HRT led to high nitrogen removal performance (91.4 %–96.3 %) and low N2O emission factor (2.8 %). The simultaneous nitrification and denitrification (SND) in swing zones and endogenous denitrification in anoxic zones contributed to the nitrogen removal. Meanwhile, the SND and autotrophic denitrification processes were identified as the N2O sources. Low DO level enriched ammonia-oxidizing bacteria and enhanced the SND and autotrophic denitrification pathway. These findings suggest that the ASAO system is promising for reducing carbon emissions in municipal wastewater treatment.

Transition from sulfur autotrophic to mixotrophic denitrification: Performance with different carbon sources, microbial community and artificial neural network modeling

To address the limitations inherent in both sulfur autotrophic denitrification (SAD) and heterotrophic denitrification (HD) processes, this study introduces a novel approach. Three carbon sources (glucose, methanol, and sodium acetate) were fed into the SAD system to facilitate the transition towards mixotrophic denitrification. Batch experiments were conducted to explore the effects of influencing factors (pH, HRT) on the denitrification performance of the mixotrophic system. Carbon source dosages were varied at 12.5%, 25%, and 50% of the theoretical amounts required for HD (18, 36, and 72 mg/L, respectively). The results showed distinct optimal dosages for each of the three organic carbon sources. The mixotrophic system, initiated with sodium acetate at 25% of the theoretical value, demonstrated the highest denitrification performance, achieving NO3−-N removal efficiency of 99.8% and the NRR of 6.25 mg/(L·h). In contrast, the corresponding systems utilizing glucose (at 25% of the theoretical value) and methanol (at 50% of the theoretical value) achieved lower removal efficiency of 77.0% and 88.4%, respectively. The corresponding NRRs were 4.85 mg/(L·h) and 5.65 mg/(L·h). Following the transition from SAD to a mixotrophic system, the abundance of Thiobacillus decreased from 78.5% to 34.4% at the genus level, and the mixotrophic system cultivated a variety of other denitrifying bacteria (Thauera, Aquimonas, Azoarcus, and Pseudomonas), indicating an enhanced microbial community structure diversity. The established artificial neural network (ANN) model accurately predicted the effluent quality of mixotrophic denitrification, which predicted values closely aligning with experimental results (R2 > 0.9). Furthermore, initial pH exerted greater relative importance for COD removal and sulfur conversion, while the relative importance of HRT was more pronounced for NO3−-N removal.

Nutrient removal performance and the nitrogen-sulfur conversion pathways in sulfur-iron based biofilter under acidic/alkaline conditions

The nitrate and phosphate removal in sulfur based (Vp) and sulfur-iron based autotrophic denitrification systems (Sp) with sponge iron as the iron source was investigated under different pH (6–9). In this study, Sp (>98 %) and Vp (>97 %) had higher nitrate removal efficiency at pH 7–8 and the highest phosphate removal efficiency (94.9 % ± 3.57 %) at pH 6. The flow cytometry detection revealed that the cell integrity was damaged due to the formation of iron crusts at pH 6, which resulted in irreversible inhibition in Sp. Electron transfer and nitrogen transformation showed that compared with the sustained inhibition of denitrification at pH 6, the inhibition at pH 9 occurred mainly in the initial 0–2 h due to the enhanced flow of electrons toward the formation of biological element sulfur (Sbio0). Coupled sponge iron promoted the adsorption and reduction of NO2−/N2O and reduced N2O emissions. Metatranscriptomics analysis showed that significant upregulation of key gene expression for nitrogen and sulfur metabolism in Sp, with complex regulatory mechanisms enhancing system stability and performance. Acidic conditions reduced the abundance of denitrification and sulfur oxidation functional genes. However, alkaline conditions (pH of 9) mainly caused the decreased abundance of denitrification functional genes, which was favorable for accumulation of Sbio0. This study provides a theoretical basis for the stable operation and regulation of the sulfur-iron based autotrophic denitrification process.

A novel sulfur-based fiber carrier fixed-bed reactor for nitrate-contaminated wastewater treatment: Performance, operational characteristics and cold-tolerant mechanism

In the view of the serious prejudice of nitrates, sulfur-based autotrophic denitrification filter (SADF) has been widely used for deep nitrate removal from electron-donor-deficient water. However, it faces many challenges like slow start-up, reliance on backwashing, and poor low-temperature tolerance. For these challenges, a novel sulfur-based fiber carrier fixed bed reactor (SFFR) was developed in this study. It was found that SFFR had great film-forming ability and flow field characteristics, which promoted it to obtain superior denitrification performance, and the maximum nitrogen removal rate of 0.53 kg-N/m3/d. Meanwhile, SFFR shows significant cold tolerance, and alleviating the acidification problem of effluent to a certain extent. To sum up, the SFFR possesses the potential to be widely used in real-world wastewater treatment applications, especially as a promising solution for nitrate removal in cold regions. This study can provide an important reference for the improvement of the elemental sulfur autotrophic denitrification process.

Comparative analysis of sulfur-driven autotrophic denitrification for pilot-scale application: Pollutant removal performance and metagenomic function

Two parallel pilot-scale reactors were operated to investigate pollutant removal performance and metabolic pathways in elemental sulfur-driven autotrophic denitrification (SDAD) process under low temperature and after addition of external electron donors. The results showed that low temperature slightly inhibited SDAD (average total nitrogen removal of ∼4.7 mg L−1) while supplement of sodium thiosulfate (stage 2) and sodium acetate (stage 3) enhanced denitrification and secretion of extracellular polymeric substances (EPS), leading to the average removal rate of 0.75 and 1.01 kg N m−3 d−1, respectively with over twice higher total EPS. Correspondingly, nitrogen and sulfur related microbial metabolisms especially nitrite reductase and nitric oxide reductase encoding were promoted by genera including Thermomonas and Thiobacillus. The variations revealed that extra sodium acetate improved denitrification and enriched more SDAD-related microorganisms compared with sodium thiosulfate, which potentially catalyzed the refinement of practical strategies for optimizing denitrification in low carbon to nitrogen ratio wastewater treatment.

Impact of copper and sulfamethazine stress on microbial community and antibiotic resistance genes in denitrification systems: Comparison between heterotrophic and sulfur autotrophic denitrification processes

As common pollutants in wastewater, heavy metals and antibiotics have negative effects on the performance, microbial community and antibiotic resistance genes (ARGs) propagation of biological treatment processes. This study investigated the differential responses of heterotrophic denitrification (HD) and sulfur autotrophic denitrification (SAD) process to Cu(II) and sulfamethazine (SMZ) stress. Exposure to 12 mg/L Cu(II) and 25 mg/L SMZ, either alone or combined, reduced nitrogen removal efficiency by 4.22 % to 32.6 % for HD and 38.7 % to 63.2 % for SAD, respectively. HD exhibited greater robustness and microbial cooperation compared to SAD under stress. Cu(II) and SMZ intensified bacterial reactive oxygen species (ROS) responses, inhibited cell activity, and reduced extracellular polymeric substance secretion. Interestingly, microbially reduced sulfides (e.g., H2S and S2−) in SAD alleviated the cellular oxidative stress induced by Cu(II) and SMZ through ROS neutralization and CuS precipitation. Under stress, the relative abundance of ARGs increased significantly more in HD than in SAD compared to their respective control groups. Network analysis revealed more potential ARGs hosts in HD than in SAD. This study underscored the potential of the microbially reduced sulfides generated in the SAD process to effectively mitigate the spread of ARGs under selective pressure from heavy metals and antibiotics.

Unraveling the impact of perfluorooctanoic acid on sulfur-based autotrophic denitrification process

PFOA has garnered heightened scrutiny for its impact on denitrification, especially given its frequent detection in secondary effluent discharged from wastewater treatment plants. However, it is still unclear what potential risk PFOA release poses to a typical advanced treatment process, especially the sulfur-based autotrophic denitrification (SAD) process. In this study, different PFOA concentration were tested to explore their impact on denitrification kinetics and microbial dynamic responses of the SAD process. The results showed that an increase PFOA concentration from 0 to 1000 μg/L resulted in a decrease in nitrate removal rate from 9.52 to 7.73 mg-N/L·h. At the same time, it increased nitrite accumulation and N2O emission by 6.11 and 2.03 times, respectively. The inhibitory effect of PFOA on nitrate and nitrite reductase activity in the SAD process was linked to the observed fluctuations in nitrate and nitrite levels. It is noteworthy that nitrite reductase was more vulnerable to the influence of PFOA than nitrate reductase. Furthermore, PFOA showed a significant impact on gene expression and microbial community. Metabolic function prediction revealed a notable decrease in nitrogen metabolism and an increase in sulfur metabolism under PFOA exposure. This study highlights that PFOA has a considerable inhibitory effect on SAD performance.

Effect of elemental sulfur on anaerobic ammonia oxidation: Performance and mechanism

Biological nitrogen removal processes provide effective means to mitigate nitrogen-related issues in wastewater treatment. Previous studies have highlighted the collaborative efficiency between sulfur autotrophic denitrification and Anammox processes. However, the trigger point induced the combination of nitrogen and sulfur metabolism is unclear. In this study, elemental sulfur (S0) was introduced to Anammox system to figure out the performance and mechanism of S0-mediated autotrophic denitrification and Anammox (S0SAD-A) systems. The results showed that the nitrogen removal performance of the Anammox reactor decreased with the increasing concentrations of NH4+-N and NO2−-N in influent, denitrification occurred when NH4+-N concentration reached 100 mg/L. At stage ⅳ (150 mg/L NH4+-N), the total nitrogen removal efficiency in S0SAD-A system (95.99%) was significantly higher than that in the Anammox system (77.22%). Throughout a hydraulic retention time, the consumption rate of NH4+-N in S0SAD-A was faster than that in Anammox reactor. And there existed a nitrate-concentration peak in S0SAD-A system. Metagenomic sequencing was performed to reveal functional microbes as well as key genes involved in sulfur and nitrogen metabolism. The results showed that the introduction of S0 elevated the abundance of Ca. Brocadia. Moreover, the relative abundance of Anammox genes, such as hao, hzsA and hzsC were also stimulated by sulfur. Notably, unclassified members in Rhodocyclaceae acted as the primary contributor to key genes involved in the sulfur metabolism. Overall, the interactions between Anammox and denitrification were stimulated by sulfur metabolism. Our study shed light on the potential significance of Rhodocyclaceae members in the S0SAD-A process and disclosed the relationship between anammox and denitrification.

The stratified response of heterotrophic, autotrophic or mixotrophic denitrification to enrofloxacin stress along different filling heights in up-flow fixed-bed bioreactors

Antibiotics frequently exist in nitrate wastewater and seriously threaten biological denitrification. This study investigated the impact of enrofloxacin (ENR) on autotrophic, heterotrophic, and mixotrophic denitrification processes at various filling heights. The experiments were conducted across four consecutive phases with ENR concentrations of 0, 0.1, 1, and 10 mg/L. As the influent ENR concentration increased, the denitrification performance of the FeS2-based autotrophic denitrification (PAD) system was significantly inhibited at different filling heights, with nitrate removal efficiency (NrE) dropping to 30.42 %. In contrast, the polycaprolactone (PCL)-based heterotrophic denitrification (PHD) and mixotrophic denitrification (PAD+PHD) systems only affected NrE in the lower layer, whereas the middle and upper layers maintained high NrE levels, largely unaffected by ENR stress, reaching up to 98.28 % and 94.02 % respectively. Sulfate reduction was more prominent in the upper and middle layers of the mixotrophic denitrification bioreactor (PHD system). Bacterial diversity indices initially increased and then decreased as ENR concentration rose from 0 to 10 mg/L, with the PHD system showing lower diversity compared to the PAD system. Redundancy analysis (RDA) revealed that the relative abundances of Proteobacteria and Actinobacteriota in the PAD system, and Bacteroidota and Firmicutes in the PHD system, were negatively correlated with ENR concentration. Overall, the PAD system experienced significant stress from ENR at various filling heights, whereas the upper-middle layer of the PHD system demonstrated greater resistance. This highlighted the impact of antibiotic contamination along with the performance of different filling heights and emphasized the need for tailored strategies in wastewater treatment.

Effective remediation of agricultural drainage at three influent strengths by bioaugmented constructed wetlands filled with mixture of iron‑carbon and organic solid substrates: Performance and mechanisms

Agricultural drainage containing a large quantity of nutrients can cause quality deterioration and algal blooming of receiving water bodies, thus needs to be effectively remediated. In this study, iron‑carbon (FeC) composite-filled constructed wetlands (Fe-C-CWs) were employed to treat farmland drainage at three pollution levels, and organic solid substrates (walnut shells) and phosphate-accumulating denitrifying bacteria (Pseudomonas sp. DWP1) were supplemented to enhance the treatment performance. The results showed that the Fe-C-CWs exhibited notably superior removal efficiency for total nitrogen (TN, 52.0–58.2 %), total phosphorus (TP, 67.8–70.2 %) and chemical oxygen demand (COD, 56.7–70.4 %) than the control systems filled solely with gravel (28.5–32.5 % for TN, 33.2–40.5 % for TP and 30.2–55.0 % for COD) at all influent strengths, through driving autotrophic denitrification, Fe-based dephosphorization, and organic degradation processes. The addition of organic substrates and functional bacteria markedly enhanced pollutant removal in the Fe-C-CWs. Furthermore, use of FeC and organic substrates and denitrifier inoculation decreased CO2 and CH4 emissions from the CWs, and reduced global warming potential of the CWs at low influent strength. Pollutant removal efficiencies in the CWs were only marginally impacted by the increasing influent loads except for NO3−-N, and pollutant removal mass was largely increased with the increase of influent strengths. The microbial community in the FeC composite-filled CWs exhibited distinct distribution patterns compared to the gravel-filled CWs regardless of the influent strengths, with obviously higher proportions of dominant genera Trichococcus, Geobacter and Ferritrophicum. Keystone taxa associated with pollutant removal in the Fe-C-filled CWs were identified to be Pseudomonas, Geobacter, Ferritrophicum, Denitratisoma and Sediminibacterium. The developed augmented Fe-C-filled CWs show great promises for remediating agricultural drainage with varied pollutant loads.

Inhibition of zinc ions in sulfur-driven autotrophic denitrification process: What is the behavior of extracellular polymeric substances?

Sulfur-driven autotrophic denitrification (SAD) process is a cost-effective and sustainable method for nitrogen removal from wastewater. However, a higher concentration of zinc ions (Zn(II)) flowing into wastewater treatment plants poses a potential threat to the SAD process. This study examined that a half maximal inhibitory concentration (IC50) of Zn(II) was 7 mg·L−1 in the SAD process. Additionally, the addition of 20 mg·L−1 Zn(II) resulted in a severe accumulation of nitrite to 150.20 ± 6.00 mg·L−1 when the initial concentration of nitrate was 500 mg·L−1. Moreover, the activities of nitrate reductase, nitrite reductase, dehydrogenase and electron transport system were significantly inhibited under Zn(II) stress. The addition of Zn(II) inhibited EPS secretion and worsened electrochemical properties. The result was attributed to the spontaneous binding between EPS and Zn(II), with a ΔG of −17.50 KJ·mol−1 and a binding constant of 1.77 × 104 M−1, respectively. Meanwhile, the protein, fulvic acid, and humic-like substances occurred static quenching after Zn(II) addition, with -OH and -C=O groups providing binding sites. The binding sequence was fulvic acid→protein→humic acid and -OH → -C=O. Zn(II) also reduced the content of α-helix, which was unfavorable for electron transfer. Additionally, the Zn(II) loosened protein structure, resulting in a 50 % decrease in α-helix/(β-sheet+random coil). This study reveals the effect of Zn(II) on the SAD process and enhances our understanding of EPS behavior under metal ions stress.

The impact of benzoic acid and lactic acid on the treatment efficiency and microbial community in the sulfur autotrophic denitrification process.

Nitrate poses a potential threat to aquatic ecosystems. This study focuses on the sulfur autotrophic denitrification mechanism in the process of water culture wastewater treatment, which has been successfully applied to the degradation of nitrogen in water culture farm effluents. However, the coexistence of organic acids in the treatment process is a common environmental challenge, significantly affecting the activity of denitrifying bacteria. This paper aims to explore the effects of adding benzoic acid and lactic acid on denitrification performance, organic acid removal rate, and microbial population abundance in sulfur autotrophic denitrification systems under optimal operating conditions, sulfur deficiency, and high hydraulic load. In experiments with 50 mg·L-1 of benzoic acid or lactic acid alone, the results show that benzoic acid and lactic acid have a stimulating effect on denitrification activity, with the stimulating effect significantly greater than the inhibitory effect. Under optimal operating conditions, the average denitrification rate of the system remained above 99%; under S/N = 1.5 conditions, the average denitrification rate increased from 88.34% to 91.93% and 85.91%; under HRT = 6h conditions, the average denitrification rate increased from 75.25% to 97.79% and 96.58%. In addition, the addition of organic acids led to a decrease in microbial population abundance. At the phylum level, Proteobacteria has always been the dominant bacterial genus, and its relative abundance significantly increased after the addition of benzoic acid, from 40.2% to 61.5% and 62.4%. At the genus level, Thiobacillus, Sulfurimonas, Chryseobacterium, and Thermomonas maintained high population abundances under different conditions. PRACTITIONER POINTS: Employing autotrophic denitrification process for treating high-nitrate wastewater. Utilizing organic acids as external carbon sources. Denitrifying bacteria demonstrate high utilization efficiency towards organic acids. Organic acids promote denitrification more than they inhibit it. The promotion is manifested in the enhancement of activity and microbial abundance.

Prediction of the sulfur driven autotrophic denitrification process via CFD-DEM approach

Sulfur driven autotrophic denitrification is an economic and efficient means to purify the nitrogenous wastewater. The long-term experimental study is quite time-consuming and expensive. In this work, a coupled computational fluid dynamics-discrete element method (CFD-DEM) is implemented to investigate the performance of long-term operating sulfur autotrophic denitrification process. The evolution of particle accumulation and nitrate removal efficiency in the packed bed reactors with different tube diameters and sulfur particle sizes are simulated. It is found that the NO-3-N removal rates for different tube diameters are close and a wider particle size distribution is shown in the slender reactor. The small particle bed shows a better NO-3-N removal efficiency.

Effects of initial alkalinity and elemental sulfur: dolomitic limestone ratio on autotrophic denitrification of poultry wastewater

Autotrophic denitrification from elemental sulfur is an alternative to heterotrophic denitrification for substrates with a low C/N ratio. In this process, nitrogen compounds are reduced from elemental sulfur instead of carbon, as in the conventional process. However, it may be limited by the low concentration of alkalinity in these effluents. Thus, this study evaluated the performance of autotrophic denitrification of nitrified poultry effluent in four fixed bed reactors with a ratio of elemental sulfur/dolomitic limestone (1:0, 3:1, 1:1, 1:3 v/v) under conditions of the decline of initial alkalinity (1000, 800, 600, 400 and 200 mg CaCO3 L-1). Denitrification efficiencies greater than 84.8% were observed in conditions of initial alkalinity greater than 600 mg CaCO3 L-1 in the four reactors, associated with maximum yields of 445.1 mg SO4-2 L-1. The slow dissolution of dolomitic limestone may have impaired denitrification efficiency in conditions of initial alkalinity lower than 600 mg CaCO3 L-1. The autotrophic denitrification process proved viable for the treatment of nitrified effluent, and its efficiency was linked to the food substrate and the dissolution of dolomitic limestone.

Rapid conversion of heterotrophic denitrification to autotrophic denitrification in mariculture wastewater treatment: Denitrification performance and microbial communities under antibiotic stress

Mariculture wastewater, characterized by a low COD/Nitrogen (C/N) ratio and the occurrence of antibiotics, is commonly treated by a heterotrophic denitrification (HD) process with supplying substantial carbon sources. Compared to HD, sulfur-based autotrophic denitrification (AD) requires less external carbon sources and is more favorable for low-carbon wastewater treatment. However, the AD process often consumes more startup time. Thus, the present study was designed to explore a rapid startup strategy for the AD process by switching from HD to AD, treating mariculture wastewater. A rapid transformation from HD to AD was achieved by inoculating mixotrophic denitrifying sludge, reducing carbon sources and adding sulfur sources under the antibiotic sulfamethoxazole (SMX). From the functional microbial community point of view, our results showed that autotrophic bacteria exhibited greater adaptability to sulfamethoxazole than heterotrophic bacteria. During the transition from HD to AD, nitrate removal improved from 53.13 % to 68.57 %, and the secretion of tightly bound extracellular polymeric substances (TB-EPS) increased by 29.94 mg/g VSS. Meanwhile, in the heterotrophic phase, the presence of sulfate in mariculture wastewater maintained a high abundance of sulfate-reducing bacteria such as Desulfuromusa (0.8 %) and sulfur-based autotrophic denitrifying bacteria such as Sedimenticola (36.7 %) and Thiogranum (15.2 %). This increases the activity of sulfur metabolism during the heterotrophic phase and ensures a rapid transition from HD to AD. The rapid shift from HD to AD in seawater under antibiotic stress provides theoretical support for the improvement of mariculture wastewater treatment processes.

Synergistic denitrification and Mn cycle driven by sulfur autotrophy significantly enhanced nitrogen removal

Due to the low C/N and high NO3−-N content in tailwater of wastewater treatment plants, the need for further denitrification treatment attracted wide attention. To achieve deep denitrification of tailwater with low C/N and high NO3−-N, a novel sulfur-manganese carbonate ore denitrification (SMCD) biological reactor was constructed. The denitrification performance of SMCD reactor was significantly higher than that of sulfur denitrification and manganese carbonate ore denitrification reactors. Across the study, the removal efficiencies of TN and NO3−-N were above 90%, and without NO2−-N accumulation in the SMCD reactor. Quantitative results of functional genes showed that SMCD reactor were conducive to realize complete denitrification. Besides, manganese autotrophic denitrification (MAD) process occurred in the SMCD reactor. By analyzing the nitrogen as well as SO42- and Mn(Ⅱ) concentrations along the height, it was found that the conversion between Mn(II) and Mn(IV) was stable during Stage Ⅲ (C/N=1). The microbiota in the SMCD reactor was distinctly different from that in the controls. Thiobacillus, Thermomonas, and an unclassified member in family cvE6 were the dominant members of SMCD reactor. Besides, Thermomonas, Dokdonella, and Simplicispira were identified as potential Mn oxidizers. The SMCD reactor exhibited a significant synergistic denitrification effect by coupling sulfur autotrophic denitrification and MAD process, which provided an effective solution for the deep denitrification of low C/N tailwater.

Biological denitrification performance of a novel sulfur-slow-release carbon source mixed filler

Long-term operation of the sulfur autotrophic denitrification (SAD) process leads to pH reduction, nitrite accumulation, a low nitrogen removal efficiency and excessive sulfate in the effluent. Two slow-release carbon source (SRC) fillers (reactors S2 and S3) consisting of different ratios of hydroxypropyl methyl cellulose, disodium fumarate, polyhydroxy fatty acid ester and Fe3O4 powder were designed to couple SAD with heterotrophic denitrification by using SRC and sulfur particles (reactor S1) as components, combining the advantages of both processes while avoiding secondary pollution. During reactor operation, S3 showed the best denitrification performance. Under the optimal hydraulic retention time of 4 h, different influent nitrate concentrations of 39.92 ± 0.59 mg/L and 20.22 ± 0.45 mg/L achieved average nitrate removal efficiencies up to 88.01 ± 3.62 % and 94.28 ± 2.52 %, and the average denitrification strength were 8.78 mg N/(L·h) and 4.77 mg N/(L·h), respectively. 16S rRNA gene analysis at the genus level showed that the coupled reaction had the highest relative abundance of Thiobacillus associated with SAD, and SAD dominated the coupled system.

Electrochemical-coupled sulfur-driven autotrophic denitrification for nitrogen removal from raw landfill leachate: Evaluation of performance and mechanisms

The cost-effective and environment-friendly sulfur-driven autotrophic denitrification (SdAD) process has drawn significant attention for advanced nitrogen removal from low carbon-to-nitrogen (C/N) ratio wastewater in recent years. However, achieving efficient nitrogen removal and maintaining system stability of SdAD process in treating low C/N landfill leachate treatment have been a major challenge. In this study, a novel electrochemical-coupled sulfur-driven autotrophic denitrification (ESdAD) system was developed and compared with SdAD system through a long-term continuous study. Superior nitrogen removal performance (removal efficiency of 89.1 ± 2.5 %) was achieved in ESdAD system compared to SdAD process when treating raw landfill leachate (influent total nitrogen (TN) concentration of 241.7 ± 36.3 mg-N/L), and the effluent TN concentration of ESdAD bioreactor was as low as 24.8 ± 5.1 mg-N/L, which meets the discharge standard of China (< 40 mg N/L). Moreover, less sulfate production rate (1.3 ± 0.2 mg SO42−−S/mgNOx−−N vs 1.7 ± 0.2 mg SO42−−S/mgNOx−−N) and excellent pH modulation (pH of 6.9 ± 0.2 vs 5.8 ± 0.4) were also achieved in the ESdAD system compared to SdAD system. The improvement of ESdAD system performance was contributed to coexistence and interaction of heterotrophic bacteria (e.g., Rhodanobacter, Thermomonas, etc.), sulfur autotrophic bacteria (e.g., Thiobacillus, Sulfurimonas, Ignavibacterium etc.) and hydrogen autotrophic bacteria (e.g., Thauera, Comamonas, etc.) under current stimulation. In addition, microbial nitrogen metabolic activity, including functional enzyme (e.g., Nar and Nir) activities and electron transfer capacity of extracellular polymeric substances (EPS) and cytochrome c (Cyt-C), were also enhanced during current stimulation, which facilitated the nitrogen removal and maintained system stability. These findings suggested that ESdAD is an effective and eco-friendly process for advanced nitrogen removal for low C/N wastewater.