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.

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.

Performance and microbial mechanism in sulfide-driven autotrophic denitrification by different inoculation sources in face of various sulfide and sulfate stress.

To develop a reliable sulfide (S2-) autotrophic denitrification (SAD) process under S2- and SO42- salinity stresses, the biofilm performance and microbial mechanisms were comparatively studied using different inocula of activated sludge (AS) and intertidal sediment (IS). Biofilm IS enriched more denitrification genes (0.34 %) and S2- oxidation genes (0.29 %) than those with AS. Higher denitrification performance was obtained under S2- (100 mg/L) and SO42- (5-15 g/L Na2SO4) stresses, but no significantly differences were observed under levels of 0-200 mg/L S2- and 30 g/L Na2SO4. Notably, biofilm samples in SAD systems with IS still had more S2- oxidation genes at high S2- levels of 100-200 mg/L and Na2SO4 level of 30 g/L. The key functional genus Thiobacillus accumulated well at 30 g/L Na2SO4, but was strongly inhibited at 200 mg/L S2-. The findings were advantage to SAD application under sulfide and salinity stresses.

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.

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.

Enhanced denitrification by graphene oxide-modified cathode for the secondary effluent of wastewater treatment plants in three-dimensional biofilm electrode reactors.

In this study, a novel three-dimensional biofilm electrode reactor (3D-BER) with a graphene oxide (GO)-modified cathode was developed to enhance the denitrification performance of secondary effluent from wastewater treatment plants (SEWTPs). The effects of different hydraulic retention times (HRTs) and currents on the 3D-BER were explored. The results indicated that at the optimal HRT of 4 h and current of 350 mA/m2, the 3D-BER with GO-modified cathode had a higher denitrification rate (2.40 ± 0.1 mg TN/L/h) and less accumulation of intermediate products, especially with 3.34% total nitrogen (TN) molar conversion to N2O. The GO-modified cathode offered a large biocompatible specific surface area and enhanced the conductivity, which favored microbial growth and increased electron transfer efficiency and extracellular enzyme activities. Moreover, the activity of nitrite reductase increased more than that of nitrate reductase to accelerate nitrite reduction, thus facilitating the denitrification process. The proposed 3D-BER provided an effective solution to elevate tertiary denitrification in the SEWTP.

Influence of residual anaerobic granular sludge (AnGS) from anaerobically digested molasses wastewater in aerobic granular sludge reactor.

This study investigated the impact of residual anaerobic granular sludge (AnGS) from anaerobic digesters treating molasses wastewater on ammonium reduction in a downstream aerobic granular sludge (AGS) reactor. Two conditions were tested: raw (high AnGS concentration) and settled (low AnGS concentration) anaerobically digested molasses wastewaters were fed into the AGS reactor. With the introduction of raw wastewater, enhanced nitrite accumulation at 30 % and improved total inorganic nitrogen (TIN) removal at 11 % were observed compared to 1 % nitrite accumulation and 8 % TIN removal with the introduction of settled wastewater. However, AnGS adversely affected other aspects of reactor performance, increasing effluent solid content and decreasing soluble chemical oxygen demand removal efficiency from 20 % in the low AnGS condition to 11 % in the high AnGS condition. Despite the observed retention of AnGS in the reactor, no significant bioaugmentation effects on the microbial community of the AGS were observed. Aerobic granular sludge was consistently observed in both conditions. The study suggests that AnGS may act as a nucleus for granule formation, helping to maintain granule stability in a disturbed environment. This study offers a systematic understanding of the impact of AnGS on subsequent nitrogen removal process using AGS, aiding in the decision making in the treatment of high solid anaerobic digestate.

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.

Microbial communities and denitrification mechanisms of pyrite autotrophic denitrification coupled with three-dimensional biofilm electrode reactor.

Denitrification is of great significance for low C/N wastewater treatment. In this study, pyrite autotrophic denitrification (PAD) was coupled with a three-dimensional biofilm electrode reactor (BER) to enhance denitrification. The effect of current on denitrification was extensively studied. The nitrate removal of the PAD-BER increased by 14.90% and 74.64% compared to the BER and the PAD, respectively. In addition, the electron utilization, extracellular polymeric substances secretion, and denitrification enzyme activity (NaR and NiR) were enhanced in the PAD-BER. The microbial communities study displayed that Dokdonella, Hydrogenophaga, Nitrospira, and Terrimonas became the main genera for denitrification. Compared with the PAD and the BER, the abundance of the key denitrification genes narG, nirK, nirS, and nosZ were all boosted in the PAD-BER. This study indicated that the enhanced autotrophic denitrifiers and denitrification genes were responsible for the improved denitrification in the PAD-BER. PRACTITIONER POINTS: PAD-BER displayed higher nitrate removal, EPS, NAR, and NIR activity. The three types of denitrification (HD, HAD, and PAD) and their contribution percentage in the PAD-BER were analyzed. HAD was dominant among the three denitrification processes in PAD-BER. Microbial community composition and key denitrification genes were tested to reveal the denitrification mechanisms.

Microbial enhanced manganese-autotrophic denitrification in reactor: performance, microbial diversity, potential functions.

Autotrophic denitrification technology has gained increasing attention in recent years owing to its effectiveness, economical, and environmentally friendly nature. However, the sluggish reaction rate has emerged as the primary impediment to its widespread application. Herein, a bio-enhanced autotrophic denitrification reactor with modified loofah sponge (LS) immobilized microorganisms was established to achieve efficient denitrification. Under autotrophic conditions, a nitrate removal efficiency of 59.55 % (0.642 mg/L/h) and a manganese removal efficiency of 86.48 % were achieved after bio-enhance, which increased by 20.92 % and 36.34 %. The bioreactor achieved optimal performance with denitrification and manganese removal efficiencies of 99.84 % (1.09 mg/L/h) and 91.88 %. ETSA and 3D-EEM analysis reveled manganese promoting electron transfer and metabolic activity of microorganisms. High-throughput sequencing results revealed as the increase of Mn(II) concentration, Cupriavidus became one of the dominant strains in the reactor. Prediction of metabolic functions results proved the great potential for Mn(II)-autotrophic denitrification of LS bioreactor.

Intertwining of the C-N-S cycle in passive and aerated constructed wetlands.

The microbial processes occurring in constructed wetlands (CWs) are difficult to understand owing to the complex interactions occurring between a variety of substrates, microorganisms, and plants under the given physicochemical conditions. This frequently leads to very large unexplained nitrogen losses in these systems. In continuation of our findings on Anammox contributions, our research on full-scale field CWs has suggested the significant involvement of the sulfur cycle in the conventional C-N cycle occurring in wetlands, which might closely explain the nitrogen losses in these systems. This paper explored the possibility of the sulfur-driven autotrophic denitrification (SDAD) pathway in different types of CWs, shallow and deep and passive and aerated systems, by analyzing the metagenomic bacterial communities present within these CWs. The results indicate a higher abundance of SDAD bacteria (Paracoccus and Arcobacter) in deep passive systems compared to shallow systems and presence of a large number of SDAD genera (Paracoccus, Thiobacillus, Beggiatoa, Sulfurimonas, Arcobacter, and Sulfuricurvum) in aerated CWs. The bacteria belonging to the functional category of dark oxidation of sulfur compounds were found to be enriched in deep and aerated CWs hinting at the possible role of the SDAD pathway in total nitrogen removal in these systems. As a case study, the percentage nitrogen removal through SDAD pathway was calculated to be 15-20% in aerated wetlands. The presence of autotrophic pathways for nitrogen removal can prove highly beneficial in terms of reducing sludge generation and hence reducing clogging, making aerated CWs a sustainable wastewater treatment solution.

One-stage anammox and thiocyanate-driven autotrophic denitrification for simultaneous removal of thiocyanate and nitrogen: Pathway and mechanism.

The coupled process of anammox and reduced-sulfur driven autotrophic denitrification can simultaneously remove nitrogen and sulfur from wastewater, while minimizing energy consumption and sludge production. However, the research on the coupled process for removing naturally toxic thiocyanate (SCN-) is limited. This work successfully established and operated a one-stage coupled system by co-cultivating mature anammox and SCN--driven autotrophic denitrification sludge in a single reactor. In this one-stage coupled system, the average total nitrogen removal efficiency was 89.68±3.33 %, surpassing that of solo anammox (81.80±2.10 %) and SCN--driven autotrophic denitrification (85.20±1.54 %). Moreover, the average removal efficiency of SCN- reached 99.50±3.64 %, exceeding that of solo SCN--driven autotrophic denitrification (98.80±0.65 %). The results of the 15N stable isotope tracer labeling experiment revealed the respective reaction rates of anammox and denitrification as 106.38±10.37 µmol/L/h and 69.07±8.07 µmol/L/h. By analyzing metagenomic sequencing data, Thiobacillus_denitrificans was identified as the primary contributor to SCN- degradation in this coupled system. Furthermore, based on the comprehensive analysis of nitrogen and sulfur metabolic pathways, as well as the genes associated with SCN- degradation, it can be inferred that the cyanate (CNO) pathway was responsible for SCN- degradation. This work provided a deeper insight into coupling anammox with SCN--driven autotrophic denitrification in a one-stage coupled system, thereby contributing to the development of an effective approach for wastewater treatment involving both SCN- and nitrogen.

Sponge iron enriches autotrophic/aerobic denitrifying bacteria to enhance denitrification in sequencing batch reactor.

Sponge iron (SFe) coupled with a sludge system has great potential for improving biological denitrification; however, the underlying mechanism is not yet fully understood. In this study, the denitrification performance and microbial characteristics of ordinary sludge and SFe-sludge systems were investigated. Overall, the SFe-sludge reactor had faster ammonium degradation rate (94.0 %) and less nitrate accumulation (1.5-53.3 times lower) than ordinary reactor during the complete operation cycle of sequencing batch reactors. The addition of SFe increased the activities of nitrate and nitrite reductases. The total relative abundance of autotrophic denitrifying bacteria (Acidovorax, Arenimonas, etc.) in the SFe-sludge system after 38 days of operation was found to be 10.6 % higher than that in the ordinary sludge reactor. The aerobic denitrifying bacteria (Dokdonella, Phaeodactylibacter, etc.) was 5.3 % higher than ordinary sludge. The SFe-sludge system improved denitrification by enriching autotrophic/aerobic denitrifying bacteria in low carbon-to-nitrogen ratio wastewater treatment.

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.

Metagenomics and Stable Isotopes Uncover the Augmented Sulfide-Driven Autotrophic Denitrification in a Seasonally Hypoxic, Sulfate-Abundant Reservoir.

The mechanism governing sulfur cycling in nitrate reduction within sulfate-rich reservoirs during seasonal hypoxic conditions remains poorly understood. This study employs nitrogen and oxygen isotope fractionation in nitrate, along with metagenomic sequencing to elucidate the intricacies of the coupled sulfur oxidation and nitrate reduction process in the water column. In the Aha reservoir, a typical seasonally stratified water body, we observed the coexistence of denitrification, bacterial sulfide oxidation, and bacterial sulfate reduction in hypoxic conditions. This is substantiated by the presence of abundant N/S-related genes (nosZ and aprAB/dsrAB) and fluctuations in N/S species. The lower 15εNO3/18εNO3 ratio (0.60) observed in this study, compared to heterotrophic denitrification, strongly supports the occurrence of sulfur-driven denitrification. Furthermore, we found a robust positive correlation between the metabolic potential of bacterial sulfide oxidation and denitrification (p < 0.05), emphasizing the role of sulfide produced via sulfate reduction in enhancing denitrification. Sulfide-driven denitrification relied on ∑S2- as the primary electron donor preferentially oxidized by denitrification. The pivotal genus, Sulfuritalea, emerged as a central player in both denitrification and sulfide oxidation processes in hypoxic water bodies. Our study provides compelling evidence that sulfides assume a critical role in regulating denitrification in hypoxic water within an ecosystem where their contribution to the overall nitrogen cycle was previously underestimated.

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 = 6 h 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.

Optimization of "sulfur-iron-nitrogen" cycle in constructed wetlands by adjusting siderite/sulfur (Fe/S) ratio.

Sulfur-siderite autotrophic denitrification (SSAD) has been proved to solve the key problem of low nitrogen removal efficiency caused by the shortage of carbon source in constructed wetlands (CWs). In this study, five vertical flow constructed wetlands (VFCWs) were constructed with different Fe/S ratios (0/0, 0/1, 1/1, 2/1 and 1/2) to optimizing SSAD process, labeled S.0, S.1, S.2, S.3 and S.4. The results showed that the best NO3--N and TN removal rates were achieved with a Fe/S ratio of 2:1 (S.3), which were 96.26 ± 1.40% and 93.63 ± 3.12%, respectively. The abundance of denitrification genes (nirS, nirK and nosZ) in S.3 was significantly increased. Illumina high-throughput sequencing analysis indicated that the abundance and diversity of microorganisms involved in the "Sulfur-Iron-Nitrogen" cycle were enriched in S.3. The current study provided that the "Sulfur-Iron-Nitrogen" cycle in CWs was optimized by adjusting Fe/S ratio, and more types of denitrifying bacteria could be enriched, thereby enhancing nitrogen removal.

Achieving Long-Term Stability of Partial Nitrification and Autotrophic Denitrification in an MABR via Sulfide Dosing.

While partial nitrification (PN) has the potential to reduce energy for aeration, it has proven to be unstable when treating low-strength wastewater. This study introduces an innovative combined strategy incorporating a low rate of oxygen supply, pH control, and sulfide addition to selectively inhibit nitrite-oxidizing bacteria (NOB). This strategy led to a stable PN in a laboratory-scale membrane aerated biofilm reactor (MABR). Over a period of 260 days, the nitrite accumulation ratio exceeded 60% when treating synthetic sewage containing 50 mg NH4+-N/L. Through in situ activity testing and high-throughput sequencing, the combined strategy led to low levels of nitrite-oxidation activity (<5.5 mg N/m2 h), Nitrospira species (relative abundance <1%), and transcription of nitrite-oxidation genes (undetectable). The addition of sulfide led to simultaneous PN and autotrophic denitrification in the single-stage MABR, resulting in over 60% total inorganic nitrogen removal. Sulfur-based autotrophic denitrification consumed nitrite and inhibited NOB conversion of nitrite to nitrate. The combined strategy has potential to be applied in large-scale sewage treatment and deserves further exploration.

Investigation of pollutants accumulation in the submerged zone for pyrite-based bioretention facilities under continuous rainfall events.

Submerged zone in bioretention facilities for stormwater treatment has been approved to be an effective structure amendment to improve denitrification capability. However, the role and influence of water quality changes in the submerged zone under natural continuous random rainfall patterns are still not clear, especially when the rainfall is less than the pore water in the submerged zone. In this study, continuous rainfall events with different rainfall volume (light rain-light rain-heavy rain) were designed in a lab-scale woodchip mulched pyrite bioretention facility to test the effects of rainfall pattern. The results exhibited that light rain events significantly affected the pollutant removal performance of bioretention for the next rainfall. Different effects were observed during the long-term operation. In the 5th month, light rain reduced the ammonia removal efficiency of subsequent rainstorm events by 8.70%, while in the 12th month, when nitrate leakage occurred, light rain led to a 40.24% reduction in the next heavy rain event's nitrate removal efficiency. Additionally, light rain would also affect the concentration of by-products in the next rainfall. Following a light rain, the concentration of sulfate in the subsequent light rainfall can increase by 24.4 mg/L, and by 11.92 mg/L in a heavy rain. The water quality in the submerged zone and media characteristics analysis suggested that nitrogen conversion capacity of the substrate and microbes, such as Nitrospira (2.86%) and Thiobacillus (35.71%), as well as the in-situ accumulation of pollutants under light rain played important roles. This study clarifies the relationship between successive rainfall events and provides a more comprehensive understanding of bioretention facilities. This is beneficial for field study of bioretention facilities in the face of complex rainfall events.

Iron-based mixotrophic denitrification for enhancing nitrate removal from municipal secondary effluent: Performance, microbial community dynamics, and economic feasibility.

High nitrate content limits the recycling of the secondary effluent of wastewater treatment plants. In the research, one biomass-iron mixture (BIM) filter material based on mixotrophic denitrification mode (heterotrophic and iron-driven autotrophic denitrification) was developed and used to construct a novel denitrification biological filter (BIM-DNBF) for the nitrogen removal of secondary effluent. BIM-DNBF had a short start-up time (approximately 9 days), and high total nitrogen removal (81 %-89 %) without external addition of organic carbon sources during the whole operation. The coexistence of dominant heterotrophic-denitrification-like Pseudomonas and Erysipelothrix as well as iron-driven autotrophic-denitrification-like Citrobacter, Acidovorax, etc. were found in the BIM-DNBF. Moreover, biomass was recognized as one key player in promoting the reduction of Fe3+ to Fe2+, thereby facilitating the occurrence of iron-driven autotrophic denitrification. In addition, BIM-DNBF was assessed to be affordable. These findings provide evidence that BIM-DNBF can be an efficient technology for nitrogen removal of secondary effluent.