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.

Thermodynamic Inhibition of Microbial Sulfur Disproportionation in a Multisubunit Designed Sulfur-Siderite Packed Bioreactor.

Sulfur disproportionation (S0DP) poses a challenge to the robust application of sulfur autotrophic denitrification due to unpredictable sulfide production, which risks the safety of downstream ecosystems. This study explored the S0DP occurrence boundaries with nitrate loading and temperature effects. The boundary values increased with the increase in temperature, exhibiting below 0.15 and 0.53 kg-N/m3/d of nitrate loading at 20 and 30 °C, respectively. A pilot-scale sulfur-siderite packed bioreactor (150 m3/d treatment capacity) was optimally designed with multiple subunits to dynamically distribute the loading of sulfur-heterologous electron acceptors. Operating two active and one standby subunit achieved an effective denitrification rate of 0.31 kg-N/m3/d at 20 °C. For the standby subunit, involving oxygen by aeration effectively transformed the facultative S0DP functional community from S0DP metabolism to aerobic respiration, but with enormous sulfur consumption resulting in ongoing sulfate production of over 3000 mg/L. Meanwhile, acidification by the sulfur oxidation process could reduce the pH to as low as 2.5, which evaluated the Gibbs free energy (ΔG) of the S0DP reaction to +2.56 kJ, thermodynamically suppressing the S0DP occurrence. Therefore, a multisubunit design along with S0DP inhibition strategies of short-term aeration and long-term acidification is suggested for managing S0DP in various practical sulfur-packed bioreactors.

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.

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.

Advancing the treatment of low carbon-to-nitrogen ratio municipal wastewater using a novel microaerobic sludge bed approach: Insights into enhanced performance and functional microbial community.

Microaerobic sludge bed systems could align with low-energy, reasonable carbon-nitrogen (C/N) ratio, and synchronous removal objectives during wastewater treatment. However, its ability to treat municipal wastewater (MW) with varying low C/N ratio, low NH4+ concentration, along with managing sludge bulking and loss are still unclear. Against this backdrop, this study investigated the performance of an Upflow Microaerobic Sludge Bed Reactor (UMSR) treating MW characterized by varying low C/N ratios and low NH4+ concentrations. The study also thoroughly examined associated sludge bulking and loss, pollutant removal efficiencies, sludge settleability, microbial community structures, functional gene variations, and metabolic pathways. Findings revealed that the effluent NH4+-N concentration gradually decreased to 0 mg/L with a decrease in the C/N ratio, whereas the effluent COD was unaffected by the influent, maintaining a concentration below 50 mg/L. Notably, TN removal efficiency reached 90% when C/N ratio was 3. The decrease in the C/N ratio (C/N ratio was 1) increased microbial community diversity, with abundances of AOB, AnAOB, aerobic denitrifying bacteria, and anaerobic digestion bacteria reaching 8.34%, 0.96%, 5.07%, and 9.01%, respectively. Microorganisms' metabolic pathways significantly shifted, showing increased carbohydrate and cofactor/vitamin metabolism and decreased amino acid metabolism and xenobiotic biodegradation. This study not only provides a solution for the effluent of different pre-capture carbon processes but also demonstrates the UMSR's capability in managing low C/N ratio municipal wastewater and emphasizes the critical role of microbial community adjustments and functional gene variations in enhancing nitrogen removal efficiency.

Remediation of arsenic- and nitrate-contaminated groundwater through iron-dependent autotrophic denitrifying culture.

Groundwater contamination with arsenic and nitrate poses a pressing concern for the safety of local communities. Bioremediation, utilizing Fe(II)-oxidizing nitrate reducing bacteria, shows promise as a solution to this problem. However, the relatively weak environmental adaptability of a single bacterium hampers practical application. Therefore, this study explored the feasibility and characteristics of a mixed iron-dependent autotrophic denitrifying (IDAD) culture for effectively removing arsenic and nitrate from synthetic groundwater. The IDAD biosystem exhibited stable performace and arsenic resistance, even at a high As(III) concentration of 800 µg/L. Although the nitrogen removal efficiency of the IDAD biosystem decreased from 71.4% to 64.7% in this case, the arsenic concentration in the effluent remained below the standard (10 µg/L) set by WHO. The crystallinity of the lepidocrocite produced by the IDAD culture decreased with increasing arsenic concentration, but the relative abundance of the key iron-oxidizing bacteria norank_f_Gallionellaceae in the culture showed an opposite trend. Metagenomic analysis revealed that the IDAD culture possess arsenic detoxification pathways, including redox, methylation, and efflux of arsenic, which enable it to mitigate the adverse impact of arsenic stress. This study provides theoretical understanding and technical support for the remediation of arsenic and nitrate-contaminated groundwater using the IDAD culture.

Optimization of operating parameters and microbiological mechanism of a low C/N wastewater treatment system dominated by iron-dependent autotrophic denitrification.

Ferrous iron (Fe2+) reduces the amount of external carbon source used for the denitrification of low-C/N wastewater. The effects of key operating parameters on the efficiency of ferrous-dependent autotrophic denitrification (FDAD) and the functioning mechanism of the microbiome can provide a regulatory strategy for improving the denitrification efficiency of low C/N wastewater. In this study, the response surface method (RSM) was used to explore the influence of four important parameters-the molar ratio of Fe2+ to NO3--N (Fe/N), total organic carbon (TOC), the molar ratio of inorganic carbon to NO3--N (IC/N) and sludge volume (SV, %)-on the FDAD efficiency. Functional prediction and molecular ecological networks based on high-throughputs sequencing techniques were used to explore changes in the structure, function, and biomarkers of the sludge microbial community. The results showed that Fe/N and TOC were the main parameters affecting FDAD efficiency. Higher concentrations of TOC and high Fe/N ratios provided more electron donors and improved denitrification efficiency, but weakened the importance of biomarkers (Rhodanobacter, Thermomonas, Comamonas, Thauera, Geothrix and unclassified genus of family Gallionellaceae) in the sludge ecological network. When Fe/N > 4, the denitrification efficiency fluctuated significantly. Functional prediction results indicated that genes that dominated N2O and NO reduction and the genes that dominated Fe2+ transport showed a slight decrease in abundance at high Fe/N levels. In light of these findings, we recommend the following optimization ranges of parameters: Fe/N (3.5-4); TOC/N (0.36-0.42); IC/N (3.5-4); and SV (approximately 35%).

Sulfur-manganese carbonate composite autotrophic denitrification: nitrogen removal performance and biochemistry mechanism.

A novel composite sulfur-manganese carbonate autotrophic denitrification (SMAD) system was developed to reduce sulfate production and provide pH buffer function while improving denitrification efficiency without external organics. The average removal efficiency of total nitrogen (TN) was 98.09% and 96.29%, and that of NO3--N was 99.53% and 97.77%, respectively, in the SMAD system with a hydraulic retention time (HRT) of 6 h and 3 h. They were significantly higher than that in the controls (quartz sand, manganese carbonate ore, and sulfur systems). The H+ produced by the sulfur autotrophic denitrification (SAD) process promoted the release of Mn2+ in the SMAD system. And this system had a stable pH with no accumulation of NO2--N. The decrease of sulfate and formation of Mn oxides through Mn2+ electron donation confirmed the presence of the manganese autotrophic denitrification (MAD) process in the SMAD system. Dominant functional bacteria in the SMAD system were Thiobacillus, Chlorobium, and Sulfurimonas, which were linked to nitrogen, sulfur, and manganese conversion and promoted denitrification. Meanwhile, Flavobacterium participating in Mn2+ oxidation was found only in the SMAD system. The SMAD system provided a new strategy for advanced tailwater treatment.

Enhancing nitrogen removal from wastewater in a low C/N ratio using an air-lift bio-electrochemical reactor (ALBER).

This study focuses on the development of an air-lift bio-electrochemical reactor (ALBER) with a continuous feeding regime. The objective is to enhance nitrogen removal from synthetic wastewater with a low carbon-to-nitrogen (C/N) ratio. The chemical oxygen demand (COD) and total nitrogen (TN) of the influent wastewater were 500 and 200 mg/L, respectively. The effect of four independent variables, i.e., temperature, hydraulic retention time (HRT), N-NH4+/TN ratio and current density in the range of 16-32 °C, 6-12 h, 25-75%, and 2-10 A/m2, respectively, at three levels on the bio-electrochemical reactor performance were investigated during the bio-electrochemical reactor operation. The Face Center Cube (FCC) of response surface methodology (RSM) was used for design of experiments and model of obtained data. The ALBER achieved the maximum TN removal of 73% (146 mg/l) using external voltage and zeolite/plastic medium at temperature of 16 °C, HRT of 6 h, current density of 2 A/m2 and N-NH4+/TN ratio of 75%. The results indicated that shortening the HRT from 12 to 6 h, reducing the temperature from 32 °C to 24 °C, increasing the current density from 2 to 6 A/m2 and the reduction of nitrate concentration caused an increase in the TN removal. The results indicated that the performance of air-lift bio-electrochemical for nitrogen removal could be attributed to autotrophic denitrification (AD) and simultaneous nitrification/denitrification (SND). The research findings suggest that the ALBER should be further studied for potential use in treating industrial wastewater at low temperatures.

Enhancing nitrogen removal in the partial denitrification/anammox processes for SO4- - Rich wastewater treatment: Insights into autotrophic and mixotrophic strategies.

The investigation of partial denitrification/anammox (PD/anammox) processes was conducted under autotrophic (N-S cycle) and mixotrophic (N-S-C cycle) conditions over 180 days. Key findings revealed the remarkable capability of SO42--dependent systems to produce NO2- effectively, supporting anaerobic NH4+ oxidation. Additionally, SO42- served as an additional electron acceptor in sulfate reduction ammonium oxidation (SRAO). Increasing influent SO42- concentrations notably improved ammonia utilization rates (AUR) and NH4+ and total nitrogen (TN) utilization efficiencies, peaking at 57% for SBR1 and nearly 100% for SBR2. Stoichiometric analysis showed a 7.5-fold increase in AUR (SRAO and anammox) in SBR1 following SO42- supplementation. However, the analysis for SBR2 indicated a shift towards SRAO and mixotrophic denitrification, with anammox disappearing entirely by the end of the study. Comparative assessments between SBR1 and SBR2 emphasized the impact of organic compounds (CH3COONa) on transformations within the N-S-C cycle. SBR1 performance primarily involved anammox, SRAO and other SO42- utilization pathways, with minimal S-dependent autotrophic denitrification (SDAD) involvement. In contrast, SBR2 performance encompassed SRAO, mixotrophic denitrification, and other pathways for SO42- production. The SRAO process involved two dominant genera, such as Candidatus Brocadia and PHOS-HE36.

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.

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.

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.

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.

Efficient nitrogen removal through coupling biochar with zero-valent iron by different packing modes in bioretention system.

Three kinds of bioretention were designed to explore the effects of zero-valent iron (ZVI) and biochar on the nitrogen removal performance and to seek a more reasonable packing method in this study. The results showed that the effluent removal rates of nitrate, ammonium and total nitrogen were 53.30 ± 12.68%, 98.41 ± 0.38% and 64.03 ± 8.72% respectively in Bioretention-3 during the rainfall events, while the nitrate concentration decreased gradually with the increase of drying time. According to the batch experiment, it was found that zero-valent iron could release continuously and stably in Bioretention-3 and Bioretention-1 due to the interception effect of biochar on dissolved oxygen. In addition, biochar in soil layer could protect zero-valent iron from excessive oxidation while biochar in the substrate layer could release organic matter to promote heterotrophic denitrification. Microbial community analysis showed that the dominant phyla were Proteobacteria (20.92-40.81%) and Actinobacteriota (9.89-24.54%). The dominant nitrifying genera was Nitrospira while there were also aerobic denitrifying bacteria (Sphingomonas, Bradyrhizobium and Chryseolinea, etc.) in soil layer. In the substrate layer, there was more ferrous iron-mediated autotrophic denitrification process (Thiobacillus, Geobacter and Denitratisoma, etc.) in Bioretention-1 and Bioretention-3 while a larger proportion of Dissimilatory Nitrate Reduction to Ammonium process (DNRA) (Bacillus, Desulfovibrio and Pseudomonas, etc.) in Bioretention-2. In general, this study showed that biochar addition in soil coupled with mixing zero-valent iron and biochar as substrate layer was a more stable and efficient design through various aspects of evidence. It provides a new way for how to use zero-valent iron and biochar to improve nitrogen removal capacity in stormwater management.

Iron mediated autotrophic denitrification for low C/N ratio wastewater: A review.

In recent years, iron mediated autotrophic denitrification has been a concern because it overcomes the absence of organic carbon and has been successfully used in denitrification for low C/N ratio wastewater. However, there is currently a lack of a more systematic summary of iron-based materials that can be used for denitrification, and no detailed overview about the mechanism of iron mediated autotrophic denitrification has been reported. In this study, the iron materials with different valence states that can be used for denitrification were summarized, and emphasized, as well as the mechanism in different interaction systems were emphasize. In addition, the contribution of various microorganisms in nitrate reduction were analyzed and the effects of operating conditions and water quality were evaluated. Finally, the challenges and shortcomings of the denitrification process were discussed aiming to find better practical engineering applications of iron-based denitrification.

The humic substance analogue antraquinone-2, 6-disulfonate (AQDS) enhanced zero-valent iron based autotrophic denitrification: Performances and mechanisms.

Zero-valent iron based autotrophic denitrification (ZVI-AD) has attracted increasing attentions in nitrate removal due to saving organic carbon budget in wastewater treatment, but limited by the low reaction speed, poor electron transfer efficiency as well as the compaction/blocking by iron hydrolysis products. Humic substances (HS) were promising to regulate iron cycle and accelerate electron transfer by serving as electron mediators. In this study, HS analogue, antraquinone-2, 6-disulfonate (AQDS), was added to enhance ZVI-AD process. Results showed that the dosage of AQDS led to a NO3--N removal efficiency of 83.37 ± 3.98% within 96 h, which was 32.28 ± 1.25% higher than that in ZVI-AD system. The corrosion of ZVI and microbially nitrate reduction were both improved at the presence of AQDS. The addition of AQDS enriched the functional species, including autotrophic denitrobacteria namely Thauera and Hydrogenophaga, iron redox-related species namely Ferruginibacter and HS respiration related species namely Flavobacterium. The genes napA and napB related to electron transfer, nirK and nosZ related to the accumulation of intermediate products were also enriched by the addition of AQDS. AQDS addition boosted the electrons flowing to both abiotic and biotic nitrate reduction. Nitrate removal mechanism involved in ZVI-AQDS coupled system was proposed. This study provided an alternative strategy for improving ZVI-AD by HS.

Effect of ibuprofen (IBU) on the sulfur-based and calcined pyrite-based autotrophic denitrification (SCPAD) systems with two filling modes: Performance and toxic response mechanism.

To investigate the effect of ibuprofen (IBU) on the sulfur-based and calcined pyrite-based autotrophic denitrification (SCPAD) systems, two individual reactors with the layered filling (L-SCPAD) and mixed filling (M-SCPAD) systems were established via sulfur and calcined pyrite. Effluent NO3--N concentration of the L-SCPAD and M-SCPAD systems was first increased to 6.44, 0.93 mg/L under 0.5 mg/L IBU exposure and gradually decreased to 1.66 mg/L, 0 mg/L under 4.0 mg/L IBU exposure, indicating that NO3--N removal performance of the M-SCPAD system was better than that of the L-SCPAD system. The variation of extracellular polymeric substances (EPS) characteristics demonstrated that more EPS was secreted in the M-SCPAD system compared to the L-SCPAD system, which contributed to forming a more stable biofilm structure and protecting microorganisms against the toxicity of IBU in the M-SCPAD system. Moreover, the increased electron transfer impedance and decreased cytochrome c implied that IBU inhibited the electron transfer efficiency of the L-SCPAD and M-SCPAD systems. The decreased adenosine triphosphate (ATP) and electron transfer system activity (ETSA) content showed that IBU inhibited metabolic activity, but the M-SCPAD system exhibited higher metabolic activity compared to the L-SCPAD system. In addition, the analysis of the bacterial community indicated a more stable abundance of nitrogen removal function bacteria (Bacillus) in the M-SCPAD system compared to the L-SCPAD system, which was conducive to maintaining a stable denitrification performance. The toxic response mechanism based on the biogeobattery effect was proposed in the SCPAD systems under IBU exposure. This study provided an important reference for the long-term toxic effect of IBU on the SCPAD systems.

Response of sulfur-metabolizing biofilm to external sulfide in element sulfur-based denitrification packed-bed reactor.

Dosing sulfide into the sulfur-packed-bed (S0PB) has great potential to enhance the denitrification efficiency by providing compensatory electron donors, however, the response of sulfur-metabolizing biofilm to various sulfide dosages has never been investigated. In this study, the S0PB reactor was carried out with increasing sulfide dosages by 3.6 kg/m3/d, presenting a decreasing effluent nitrate from 14.2 to 2.7 mg N/L with accelerated denitrification efficiency (k: 0.04 to 0.27). However, 6.5 mg N/L of nitrite accumulated when the sulfide dosage exceeded 0.9 kg/m3/d (optimum value). The increasing electron export contribution of sulfide a maximum of 85.5% illustrated its competition with the in-situ sulfur. Meanwhile, over-dosing sulfide caused serious biofilm expulsion with significant decreases in the total biomass, live cell population, and ATP by 90.2%, 86.7%, and 54.8%, respectively. This study verified the capacity of dosing sulfide to improve the denitrification efficiency in S0PB but alerted the negative effect of exceeded dosing.

Synergistic between autotrophic and heterotrophic microorganisms for denitrification using bio-S as electron donor.

In recent years, biological sulfur (bio-S) was employed in sulfur autotrophic denitrification (SAD) in which autotrophic Thiobacillus denitrificans and heterotrophic Stenotrophomonas maltophilia played a key role. The growth pattern of T.denitrificans and S.maltophilia exhibited a linear relationship between OD600 and CFU when OD600 < 0.06 and <0.1, respectively. When S.maltophilia has applied alone, the NorBC and NosZ were undetected, and denitrification was incomplete. The DsrA of S.maltophilia could produce sulfide as an alternative electron donor for T.denitrificans. Even though T.denitrificans had complete denitrification genes, its efficiency was low when used alone. The interaction of T.denitrificans and S.maltophilia reduced nitrite accumulation, leading to complete denitrification. A sufficient quantity of S.maltophilia may trigger the autotrophic denitrification activity of T.denitrificans. When the colony-forming units (CFU) ratio of S.maltophilia to T.denitrificans was reached at 2:1, the highest denitrification performance was achieved at 2.56 and 12.59 times higher than applied alone. This research provides a good understanding of the optimal microbial matching for the future application of bio-S.