Autotrophic Denitrification System Research Articles

Effect of magnetic powder (Fe3O4) on heterotrophic-sulfur autotrophic denitrification efficiency and electron transport system activity for marine recirculating aquacultural wastewater treatment

As an efficient nitrogen removal process, heterotrophic-sulfur autotrophic denitrification (HSAD) has attracted extensive attention in wastewater treatment. However, the effects of magnetic powder (Fe3O4) on the electron transport activity in HSAD process remain unclear. Therefore, in this study, a heterotrophic-sulfur autotrophic denitrification system was established to remove nitrogen from marine recirculating aquacultural wastewater for evaluating the effects of Fe3O4. At the optimal Fe3O4 concentration of 50 mg/L, the nitrogen removal efficiency reached 100% with lower sulfate accumulation, and the start-up time was shortened. The assays of denitrifying enzymes and electron transport system activity showed that Fe3O4 improved the activities of nitrate and nitrite reductases, and increased the efficiency of electron transport. Microbial community analysis revealed that Fe3O4 enriched heterotrophic denitrifier Thauera and sulfur autotrophic denitrifier Canditatus Thiobios, and thus enhanced denitrification efficiencies. This study demonstrated that Fe3O4 is an efficient denitrification accelerator in HSAD for treating marine recirculating aquacultural wastewater.

The construction of double short-cut sulfur autotrophic denitrification and PN/Anammox partition and its effect on the coupling performance of nitrogen and sulfur removal

How to reduce NO3--N from partial nitrification /Anammox (PN/Anammox) for low carbon emissions, thus achieve deep nitrogen removal, is a significant issue when applying PN/Anammox to treatment inorganic high-ammonia wastewater. This study designed a prepositioned double short-cut sulfur autotrophic denitrification (DSSADN) system integrated with PN/Anammox, achieving simultaneous sulfur recovery and complete autotrophic denitrification. It also examined the impact of reflux changes on the nitrogen and sulfur transformation characteristics in the coupled system when treating ammonia and sulfur-containing wastewater. Results indicate that the coupling of DSSADN zone (zone C1) enhanced TN removal by PN/Anammox, with the TN removal rate (TNRRT), TN removal efficiency (TNRET), and S2- removal rate reaching 0.54 kg/(m³·d), 95.1%, and 100%, respectively. The NO2--N accumulation efficiency (NiAEC1) of DSSADN reached 87.1%, with ΔSO42--S consistently less than 11.8 mg/L, and the S0 accumulation efficiency (S0AEC1) reaching 82.4%. A small amount of O2 carried in the reflux can effectively reduce residual S2- in zone C1, preventing its entry into the PN/Anammox zone (zone C2) and mitigating its toxic effects on Anammox. However, O2 also increases the expression of soxB in zone C1, causing more S2- to be oxidized to SO42-, and reducing S0AEC1. When DOC1 was between 0.05 mg/L and 0.34 mg/L, nirK expression was significantly increased, and the abundance of related functional microorganisms changed, led to more NO3--N being reduced to N2. When DOC1 exceeds 0.34 mg/L, it inhibits the denitrification process, leading to elevated NiAE. Therefore, when regulating reflux in DSSADN+PN/Anammox partitioned integrated coupling system, attention should be given to changes in DOC1, controlling it within 0.05 mg/L.

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.

Exploring the impact of fulvic acid on electrochemical hydrogen-driven autotrophic denitrification system: Performance, microbial characteristics and mechanism

In this study, the effect of modulating fulvic acid (FA) concentrations (0, 25 and 50 mg/L) on nitrogen removal in a bioelectrochemical hydrogen autotrophic denitrification system (BHDS) was investigated. Results showed that FA increased the nitrate (NO3–-N) removal rate of the BHDSs from 37.8 to 46.2 and 45.2 mg N/(L·d) with a current intensity of 40 mA. The metagenomic analysis revealed that R2 (25 mg/L) was predominantly populated by autotrophic denitrifying microorganisms, which enhanced denitrification performance by facilitating electron transfer. Conversely, R3 (50 mg/L) exhibited an increase in genes related to the heterotrophic process, which improved the denitrification performance through the collaborative action of both autotrophic and heterotrophic denitrification pathways. Besides, the study also identified a potential for nitrogen removal in Serpentinimonas, which have been rarely studied. The interesting set of findings provide valuable reference for optimizing BHDS for nitrogen removal and promoting specific denitrifying genera within the system.

Isotope analysis of nitrogen removal pathways and N2O production potential in the SDAD-anammox system under different N/S ratios

This study explored the impact of varying nitrate to sulfide (N/S) ratios on nitrogen removal efficiency (NRE) in the sulfide-driven autotrophic denitrification and anammox (SDAD-anammox) system. Optimal nitrogen removal was observed at N/S ratios between 1.5 and 2.0. Isotope tracing results showed that the contribution of anammox to nitrogen removal was enhanced with increasing N/S ratios, reaching up to 37 % at the N/S ratio of 2.5. Additionally, complex nitrogen pathways were identified, including dissimilatory nitrate reduction to ammonium (DNRA). Furthermore, isotope tracing was innovatively applied to investigate N2O emissions, demonstrating that higher N/S ratios significantly reduced N2O emissions, with the lowest emissions at N/S ratio of 2.5. Gene expression analysis indicated that nitrogen and sulfide transformation genes decreased with increasing N/S ratios, while anammox-related genes first increased and then decreased, reflecting the system's microbial dynamics. These findings offer insights into nitrogen transformation pathways and N2O production mechanisms in the SDAD-anammox process.

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.

New insights and enhancement mechanisms of activated carbon in autotrophic denitrification system utilizing zero-valent iron as indirect electron donors

Zero-valent iron acts as an indirect electron donor, supplying ferrous iron for the nitrate-dependent ferrous oxidation (NDFO) process. The addition of activated carbon (AC) increased the specific NDFO activity in situ and ex situ by 0.4 mg-N/(d·g VSS) and 2.2 mg-N/(d·g VSS), respectively, due to the enrichment of NDFO bacteria. Furthermore, AC reduced the nitrous oxide emission potential of the sludge, a mechanism that metagenomic analysis suggests may act as a cellular energy storage strategy. During a 196-day experiment, a total nitrogen removal efficiency of 53.7 % was achieved, which may be attributed to the upregulation of key genes involved in iron oxidation and denitrification. Based on these findings, a model involving pilin, ’nanowires,’ and a cyc2/?→/(FoxE→FoxY)/?→cymA/Complex III/?-mediated pathway for extracellular electron uptake was proposed. Overall, this work provides a feasible strategy for enhancing the nitrogen removal performance of the ZVI-NDFO process.

Sulfur-driven microbial fuel cells denitrification system for targeted treatment of nitrate-containing groundwater: Performance and mechanism

Sulfur (S0)-driven autotrophic denitrification system has obvious advantages in economy and performance in treating the low carbon/nitrogen groundwater. However, the by-products produced from S0 oxidation will cause disturbance to the water quality. In this study, a dual-chamber microbial fuel cell was driven by S0 (S0-MFC) to separate the S0 and nitrate, the electrons generated by the oxidation of S0 in the anodic chamber transferred to the cathode for nitrate reduction. Under the action of the anode microorganisms, the S0 was directly oxidized to sulfate. The maximum nitrate removal rate (NRR) in the cathodic chamber was 2.31 ± 0.03 mg N/L/h, and the anode electron utilization rate could reached to 19.69 %. Electrochemical results showed that flavin and c-type cytochrome could acted as electron mediators promote the electron transfer processes. The presence of loop current contributed to the enhancement of microbial metabolic activity, thus improving the stability of sludge and NRR. The denitrifiers (Thiobacillus and Denitratisoma) and sulfur autotrophs (Desulfocapsa and Acidithiobacillus) played an important role in the high NRR. A low-cost and pollution-free method for removing nitrate from groundwater was realized in this study.

Effects of polyvinyl chloride microplastics and benzylalkyldimethylethyl compounds on system performance, microbial community and resistance genes in sulfur autotrophic denitrification system

Benzylalkyldimethylethyl ammonium compounds (BAC) and polyvinyl chloride microplastics (PVC MPs), as the frequently detected pollutants in wastewater treatment plants (WWTPs), have attracted more concerns on their ecosystem risks. Therefore, this study investigated how the sulfur autotrophic denitrification (SAD) system responded to the single and joint stress of PVC MPs (1, 10 and 100 mg/L) and BAC (0.5, 5 and 10 mg/L). After 100 days of operation, the presence of 10 mg/L BAC led to obviously inhibitory effects on system performance and microbial metabolic activity. And the additions of PVC MPs or/and BAC stimulated the proliferation of intracellular resistance genes (RGs), whereas exposure to BAC increased the abundances of extracellular RGs and free RGs in water more significantly. Compared to the joint stress, BAC single stress resulted in higher abundances of free RGs in water, which further increased the risk of RGs propagation. Moreover, the interaction between mobile genetic elements and extracellular polymeric substances further increased the spread of RGs. Pathogens might be the potential hosts of RGs and enriched in SAD system and plastisphere, thereby leading to more serious ecological risks. This study will broaden the understanding of the environmental hazards posed by PVC MPs and BAC in WWTPs.

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.

Manganese sulfide-sulfur and limestone autotrophic denitrification system for deep and efficient nitrate removal: Feasibility, performance and mechanism

Despite the great potential of sulfur-based autotrophic denitrification, an improvement in nitrate removal rate is still needed. This study used the desulfurized products of Mn ore to develop the MnS-S0-limestone autotrophic denitrification system (MSLAD). The feasibility of MSLAD for denitrification was explored and the possible mechanism was proposed. The nitrate (100 mg/L) was almost removed within 24 h in batch experiment in MSLAD. Also, an average TN removal of 98 % (472.0 mg/L/d) at hydraulic retention time of 1.5 h in column experiment (30 mg/L) was achieved. MnS and S0 could act as coupled electron donors and show synergistic effects for nitrate removal. γ-MnS with smaller particle size and lower crystallinity was more readily utilized by the bacterium and had higher nitrate removal efficiency than that of α-MnS. Thiobacillus and Sulfurimonas were the core functional bacterium in denitrification. Therefore, MnS-S0-limestone bio-denitrification provides an efficient alternative method for nitrate removal in wastewater.

Highly effective removal of nitrate from saline wastewater by glucose-enhanced sulfur autotrophic system

The denitrification performance of the elemental sulfur autotrophic denitrification (S0AD) system requires improvements to broaden its applicability for treating nitrate (NO3−-N) wastewater contaminated with high salinity. Therefore, this work investigated the denitrification performance and mechanism of the glucose-enhanced (C:N ratio = 1) S0AD system with saline wastewater (0–15 g/L). The results showed that by adding glucose, our research successfully constructed the elemental sulfur autotrophic/heterotrophic denitrification (HS0AD) system, which showed a stable NO3−-N removal efficiency above 95 % and lower effluent sulfate generation. The NO3−-N removal efficiency of the HS0AD system was over 95.62 % with wastewater containing different salinity levels. With increasing salinity, the glucose utilization efficiency increased from 65.23 % to 74.35 %. Microbial community analysis showed that adding glucose promoted the growth of the Proteobacteria phylum and stimulated its activity under salt stress. High salinity (≥10 g/L) reduced the microbial diversity at the genus level. However, the growth of salt-tolerant denitrifying bacteria (Thiobacillus and Vitellibacter) still enabled efficient denitrification performance with the system. These microorganisms can resist high salinity stresses by releasing proteins. All results indicated that the glucose-enhanced system shows great nitrate removal potential, even under salt stress. These conclusions provide a theoretical basis for applying a synergistic system to treat saline wastewater contaminated with NO3−-N.

Effect of pyrite particle size on the denitrification performance of autotrophic or split-mixotrophic bioreactors supported by pyrite/polycaprolactone.

In this study, a pyrite-based autotrophic denitrification (PAD) system, a polycaprolactone (PCL)-supported heterotrophic denitrification (PHD) system, and a pyrite+PCL-based split-mixotrophic denitrification (PPMD) system were constructed. The pyrite particle size was controlled in 1-3, 3-5, or 5-8mm in both the PAD and PPMD systems to investigate the effect of pyrite particle size on the denitrification performance of autotrophic or split-mixotrophic bioreactors. It was found that the PAD system achieved the best denitrification efficiency with an average removal rate of 98.98% in the treatment of 1- to 3-mm particle size, whereas it was only 19.24% in the treatment of 5- to 8-mm particle size. At different phases of the whole experiment, the nitrate removal rates of both the PHD and PPMD systems remained stable at a high level (>94%). Compared with the PAD or PHD system, the PPMD system reduced the concentrations of sulfate and chemical oxygen demand in the final effluent efficiently. The interconnection network diagram explained the intrinsic metabolic pathways of nitrogen, sulfur, and carbon in the three denitrification systems at different phases. In addition, the microbial community analysis showed that the PPMD system was beneficial for the enrichment of Firmicutes. Finally, the impact mechanism of pyrite particle size on the performance of the PPMD system was proposed. PRACTITIONER POINTS: The reduction of pyrite particle size was beneficial for improving the efficiency of the PAD process. The change in particle size had an effect on NO2 --N accumulation in the PAD system. The accumulation of NH4 +-N in the PPMD system increased with the decrease in particle size. The reduction of pyrite particle size increased the production of SO4 2- in the PAD and PPMD systems. The correlations among the effluent indicators of the PAD and PPMD systems could be well explained.

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.

Impact of organic carbon on sulfide-driven autotrophic denitrification: Insights from isotope fractionation and functional genes

Additional organics are generally supplemented in the sulfide-driven autotrophic denitrification system to accelerate the denitrification rate and reduce sulfate production. In this study, different concentrations of sodium acetate (NaAc) were added to the sulfide-driven autotrophic denitrification reactor, and the S0 accumulation increased from 7.8% to 100% over a 120-day operation period. Batch experiments revealed a threefold increase in total nitrogen (TN) removal rate at an Ac−-C/N ratio of 2.8 compared to a ratio of 0.5. Addition of organic carbon accelerated denitrification rate and nitrite consumption, which shortened the emission time of N2O, but increased the N2O production rate. The lowest N2O emissions were achieved at the Ac−-C/N ratio of 1.3. Stable isotope fractionation is a powerful tool for evaluating different reaction pathways, with the 18ε/15ε values in nitrate reduction ranging from 0.5 to 1.0. This study further confirmed that isotope fractionation can reveal denitrifying nutrient types, with the 18ε (isotopic enrichment factor of oxygen)/15ε (isotopic enrichment factor of nitrogen) value approaching 1.0 for autotrophic denitrification and 0.5 for heterotrophic denitrification. Additionally, the 18ε/15ε values can indicate changes in nitrate reductase. There is a positive correlation between the 18ε/15ε values and the abundance of the functional gene napA, and a negative correlation with the abundance of the gene narG. Moreover, 18ε and 15ε were associated with changes in kinetic parameters during nitrate reduction. In summary, the combination of functional gene analysis and isotope fractionation effectively revealed the complexities of mixotrophic denitrification systems, providing insights for optimizing denitrification processes.

Effect of S/N ratio on operation characteristics and microbial community of double short-cut sulfur autotrophic denitrification system

Exploring the effects of different S/N ratios on the simultaneous accumulation of S0 and NO2−-N in the autotrophic denitrification process is of great significance for the enhanced Anammox denitrification and S0 resource recovery. The results showed that the low S/N ratio avoided the S2− accumulated toxicity, and the reactor could be quickly started within 9 days. When S/N ratio was 1, the highest S0 accumulation efficiency (S0AE) and NO2−-N accumulation efficiency (NiAE) could be obtained synchronously, which were 77.41 % and 87.83 %, respectively. The production of S0 and NO2−-N was up to 0.57 kg/(m3·d) and 0.18 kg/(m3·d), respectively. Too high or low S/N ratio would affect the S0AE and NiAE, respectively. With the increasing S/N ratio, sqr and narG genes expressions were enhanced and soxB and nirK genes expressions were weakened. Thiobacillus, Azoarcus and Pseudaminobacter were the dominant genera performing denitrification and desulphurization in double short-cut sulfur autotrophic denitrification.

Iron-based autotrophic denitrification driven by sponge iron for nitrite removal in an anaerobic bioreactor: Effect of iron and carbon source.

The study investigated the denitrification effect of the iron autotrophic denitrification process for removing nitrite under anaerobic conditions, utilizing sponge iron as the electron donor. When the C/N ratio equaled 1, defined as the ratio of chemical oxygen demand to total nitrogen (TN), and the influent nitrite nitrogen (NO2--N) was at 80 mg/L, the average steady-state TN effluent concentration of this system was 41.94 mg/L during the 79-day experiment. The TN value exhibited a significant decrease compared to both the sponge iron system (68.69 mg/L) and the carbon source system (56.50 mg/L). Sponge iron is beneficial for providing an electron donor and ensuring an anaerobic system, fostering an environment that promotes microorganism growth while effectively inhibiting the conversion of nitrite to nitrate. In addition, carbon sources play a vital role in ensuring microorganism growth and reproduction, thereby aiding in TN removal. The optimal parameters based on the effectiveness of TN removal in the iron autotrophic denitrification system were determined to be s-Fe0 dosage of 30 g/L and C/N = 1.5. These results suggest that the iron autotrophic denitrification process, driven by sponge iron, can effectively remove nitrite under anaerobic conditions.

Meta-analyzing the mechanism of pyrogenic biochar strengthens nitrogen removal performance in sulfur-driven autotrophic denitrification system: Evidence from metatranscriptomics

Sulfur-driven autotrophic denitrification (SAD) exhibits significant benefits in treating low carbon/nitrogen wastewater. This study presents an eco-friendly, cost-effective, and highly efficient method for enhancing nitrogen removal performance. The addition of biochar prepared at 300 °C (BC300) notably increased nitrogen removal efficiency by 31.60 %. BC300 concurrently enhanced electron production, the activities of the electron transfer system, and electron acceptors. With BC300, the ratio of NADH/NAD+ rose 2.00±0.11 times compared to without biochar, and the expression of NAD(P)H dehydrogenase genes was markedly up-regulated. In the electron transfer system, BC300 improved the electroactivity of extracellular polymeric substances and the activities of NADH dehydrogenase and complex III in intracellular electron transfer. Subsequently, electrons were directed into denitrification enzymes, where the nar, nir, nor, and nos related genes were highly expressed with BC300 addition. Significantly, BC300 activated the Clp and quorum sensing systems, positively influencing numerous gene expressions and microbial communication. Furthermore, the O%, H%, molar O/C, and aromaticity index in biochar were identified as crucial bioavailable parameters for enhancing nitrogen removal in the SAD process. This study not only confirms the application potential of biochar in SAD, but also advances our comprehension of its underlying mechanisms.

Deciphering the influencing mechanism of hydraulic retention time on purification performance of a mixotrophic system from the perspective of reaction kinetics.

At present, eutrophication is increasingly serious, so it is necessary to effectively reduce nitrogen and phosphorus in water bodies. In this study, a pyrite/polycaprolactone-based mixotrophic denitrification (PPMD) system using pyrite and polycaprolactone (PCL) as electron donors was developed and compared with pyrite-based autotrophic denitrification (PAD) system and PCL-based heterotrophic denitrification (PHD) system through continuous flow experiment. The removal efficiency of NO3--N (NRE) and PO43--P (PRE) and the contribution proportion of PAD in the PPMD system were significantly increased by prolonging hydraulic retention time (HRT, from 1 to 48h). When HRT was equal to 24h, the PPMD system conformed to the zero-order kinetic model, so NRE and PRE were mainly limited by the PAD process. When HRT was equal to 48h, the PPMD system met the first-order kinetic model with NRE and PRE reaching 98.9 ± 1.1% and 91.8 ± 4.5%, respectively. When HRT = 48h, the NRE and PRE by PAD system were 82.7 ± 9.1% and 88.5 ± 4.7%, respectively, but the effluent SO42- concentration was as high as 152.1 ± 13.7mg/L (the influent SO42- concentration was 49.2 ± 3.3mg/L); the NRE by PHD system was 98.5 ± 1.7%, but the PO43--P could not be removed ideally. The concentrations of NO3--N, total nitrogen, PO43--P, and SO42- in the PPMD system also showed distinct changes along the reactor column. In addition, the microbial diversity analysis showed that prolonging HRT (from 24 to 48h) increased the abundance of autotrophic denitrifying microorganisms in the PPMD system, ultimately increasing the contribution proportion of PAD.