Sulfur Autotrophic Denitrification Research Articles

Effect of dissolved oxygen on sulfur autotrophic denitrification and how to address it: an experimental and modelling work

Sulfur autotrophic denitrification (SAD) using elemental sulfur as the electron donor has aroused increasing interest of its application in treating secondary effluent from wastewater treatment plants (WWTPs). However, high influent dissolved oxygen (DO) in secondary effluent would limit the SAD process. This study examined the effect of different DO concentrations on SAD. Results revealed that both low (0-0.5 mg/L) and moderate (2.5-3.5 mg/L) DO concentrations would not harm the nitrate removal rate (NRR) (p > 0.05). However, high DO concentration (5.5-6.5 mg/L) significantly decreased the NRR (p < 0.05) through strong competition over the nitrate for electrons and cutting the relative abundance of sulfur-oxidizing bacteria (SOB). Both modeling and experimental results showed that applying internal reflux could serve as a strategy to mitigate the negative effect of high DO concentration, while keeping an appropriate ratio was crucial. When treating real membrane bioreactor (MBR) effluent with high DO concentration (5.5-6.5 mg/L), an internal reflux ratio of 0.5 boosted the NRR by 1.5 times. This study provided potential reference and strategy for dealing with high DO concentration wastewater by applying SAD technology.

Unveiling the mechanism underlying in−situ enhancement on anammox system by sulfide: Integration of biological and isotope analysis

The in−situ utilization of sulfide to remove the nitrate produced during the anaerobic ammonium oxidation (anammox) process can avoid prolonged sludge acclimatization, facilitating the rapid initiation of coupled nitrogen removal processes. However, the understanding of in−situ enhancement on anammox system by sulfide remains unclear. Herein, sulfide (Na2S) was introduced as an additional electron donor to remove the nitrate derived from the anammox under varying sulfide/nitrogen (S/N, S2−-S/NO3–-N, molar ratio) ratios (0.004–4.375). The underlying mechanisms were elucidated by molecular biology techniques including flow cytometry, quantitative polymerase chain reaction, and 16S rRNA amplicon sequencing, alongside isotope tracer analysis. Results revealed that anammox reactors, when operated with in−situ sulfide addition, exhibited a significant enhancement in total nitrogen removal efficiency (NRE) ranging from 11.5%−41.7% (achieved 96%), with the optimal S/N ratios of 0.01−0.8. Isotope tracer analysis indicated the successful coupling of the anammox, sulfur autotrophic denitrification (SADN), and dissimilatory nitrate reduction to ammonium (DNRA) processes within the system, with their contributions to nitrogen removal being 46%−50%, 24%−30%, and 20%−22%, respectively. Moreover, a notable increase in the abundance of sulfur–oxidizing bacteria (SOB) (20%−40% increase) and DNRA bacteria (10%−20% increase) were observed. Effective collaboration was further supported by the sustained viability of microbial communities. It is speculated that the heightened presence of SOB and DNRA bacteria created a low toxicity environment by converting sulfide to biogenic sulfur, thereby promoting the well−being of anammox bacteria. However, the excessive dosage of sulfide (S/N=1.8) intensified the DNRA process (contribution>35%) and weakened the anammox process, leading to an increase in effluent NH4+-N concentration and a decline in NRE. This study confirms that the in−situ adding an appropriate amount of sulfide favors achieving complete nitrogen removal in anammox system, which provides a novel avenue to resolve the issue of the residual nitrate in anammox process.

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

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

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.

Salinity responsive mechanisms of sulfur-based mixotrophic denitrification and ectoine induced tolerance enhancement

Increasing discharge of saline wastewater poses a challenge for heterotrophic and sulfur autotrophic denitrification (HSAD) in nitrogen removal. This study explored the response mechanism of mixotrophic denitrifying microorganisms to high-salt stress and feasible mitigation strategies. Results showed that denitrification efficiency was promoted at 2 % salinity but significantly decreased at 6 %, with NO3–-N removal rate decreased from 95.77 % to 38.01 %. The contribution of sulfur autotrophic denitrification (SAD) remained higher than heterotrophic denitrification (HD). High salinity stimulation led to an increase in reactive oxygen species (ROS) content, while the nicotinamide adenine dinucleotide (NADH) content, adenosine triphosphate (ATP) level, and electron transport system activity (ETSA) decreased by 10.74 %, 46.6 % and 56.28 % at 6 % salinity, respectively. Additionally, high salt inhibited the activity of nitrate reductase (NAR) and nitrite reductase (NIR), reducing the abundance of denitrifying bacteria. Notably, the addition of 250 mg/L of ectoine at 6 % salinity alleviated stress, promoted the secretion of extracellular polymeric substances (EPS) to protect the microorganisms, reduced microbial oxidative stress, and maintained metabolic and denitrification enzyme activities. Ectoine acted as an osmotic pressure regulator and protective agent, maintaining intracellular enzymes and macromolecular structure, thus enhancing denitrification efficiency. Microbial community analysis demonstrated a 3.99 % increase in HD functional bacteria abundance with ectoine addition, highlighting its crucial role in regulation community succession and stability. Structural equations modeling analysis identified the regulatory relationship paths involved in the response and mitigation of HSAD to high salinity stress. This study deepens understanding of the inhibition mechanism of HSAD caused by high-salt wastewater and provides a feasible technical solution for salt stress alleviation in sulfur-based mixotrophic denitrification.

Pyrite and Sulfur Autotrophic Denitrification for Simultaneous Nitrogen and Phosphorus Removal and Greenhouse Gas Emission Reduction Potential

Pyrite and Sulfur Autotrophic Denitrification for Simultaneous Nitrogen and Phosphorus Removal and Greenhouse Gas Emission Reduction Potential

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.

Exposure to zinc and dialkyldimethyl ammonium compound alters bacterial community structure and resistance gene levels in partial sulfur autotrophic denitrification coupled with the Anammox process

Dialkyldimethyl ammonium compound (DADMAC) is widely used in daily life as a typical disinfectant and often co-exists with the heavy metal zinc in sewage environments. This study investigated the effects of co-exposure to zinc (1 mg/L) and DADMAC (0.2–5 mg/L) on the performance, bacterial community, and resistance genes (RGs) in a partial sulfur autotrophic denitrification coupled with anaerobic ammonium oxidation (PSAD-Anammox) system in a sequencing batch moving bed biofilm reactor for 150 days. Co-exposure to zinc and low concentration (0.2 mg/L) DADMAC did not affect the nitrogen removal ability of the PASD-Anammox system, but increased the abundance and transmission risk of free RGs in water. Co-exposure to zinc and medium-to-high (2–5 mg/L) DADMAC led to fluctuations in and inhibition of nitrogen removal, which might be related to the enrichment of heterotrophic denitrifying bacteria dominated by Denitratisoma. Co-exposure to zinc and high concentration DADMAC (5 mg/L) stimulated the secretion of extracellular polymeric substances and increased the proliferation risk of intracellular RGs in sludge. This study provided insights into the application of PSAD-Anammox system and the ecological risks of wastewater containing zinc and DADMAC.

Distinct roles of biochar and pyrite substrates in enhancing nutrient and heavy metals removal in intermittent-aerated constructed wetlands: Performances and mechanism

Constructed wetlands have been widely employed as a cost-effective and environmentally friendly alternative for treating primary and secondary sewage effluents. In this study, biochar and pyrite were utilized as electron donor substrates in intermittent-aerated vertical flow constructed wetlands to strengthen the nutrient and heavy metals removal simultaneously, and the response of nutrient reduction and microbial community to heavy metals stress was also explored. The results indicated that biochar addition exhibited a better nitrogen removal, while pyrite addition greatly promoted the phosphorus removal. Moreover, the high removal efficiencies of Cu2+, Pb2+ and Cd2+ (above 90%) except for Zn2+ were obtained in each system. However, the exposure of heavy metals decreased phosphorus removal while had little effect on nitrogen removal. The influent load and intermittent aeration implementation led to a significant shift in microbial community structures, but microbial biodiversity and abundance decreased under the exposure of heavy metals. Particularly, Thiobacillus and Ferritrophicum, associated with sulfur autotrophic denitrification and iron autotrophic denitrification, were more abundant in pyrite-based wetland systems.

Sulfur autotrophic electrodialysis ion exchange membrane bioreactor (SEDIMB) for nitrate removal in water: Synergism and adaptability of electrodialysis and sulfur autotrophy

In this study, the novel sulfur autotrophic electrodialysis ion-exchange membrane bioreactor (SEDIMB) with high denitrification performance was developed without the addition of carbon source. Sulfur autotrophic denitrification occurs at the same time of nitrate enrichment by electric field to achieve harmless nitrate transformation in water. The final nitrate concentration in water chamber was 2.16 mg-N/L (removal efficiency of 94.49 %) at an initial nitrate concentration of 40 mg-N/L, with the maximum nitrate concentration in biological chamber of 4.09 mg-N/L. Nitrate removal in water chamber of the SEDIMB conformed to the first-order kinetic model (R2 = 0.98) and reaction rate constant K1 = 0.0030 (1/min). The effect of environmental variables (i.e., voltage, initial nitrate concentration and water body) on the SEDIMB denitrification performance was investigated. Increasing voltage could promote nitrate migration in water chamber, and when the voltage was 7 V, initial nitrate concentrations of 20 ∼ 60 mg-N/L was effectively removed (all > 93.54 %). Moreover, the change of water body (i.e., tap water, lake water and deionized water) did not affect denitrification performance of SEDIMB. Microbial community structure analysis indicated that Betaproteobacteria were dominant in the SEDIMB. Membrane characterizations (ATR-IR, SEM and CLSM) confirmed that the bacteria in biological chamber tended to accumulate and grow on one side of the anion exchange membrane (AEM), while could not affect the selective permeability of AEM.

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

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

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

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

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.

Salt stimulates sulfide−driven autotrophic denitrification: Microbial network and metagenomics analyses

Sulfur autotrophic denitrification (SADN) is a promising biological wastewater treatment technology for nitrogen removal, and its performance highly relies on the collective activities of the microbial community. However, the effect of salt (a prevailing characteristic of some nitrogen−containing industrial wastewaters) on the microbial community of SADN is still unclear. In this study, the response of the sulfide−SADN process to different salinities (i.e., 1.5 % salinity, 0.5 % salinity, and without salinity) as well as the involved microbial mechanisms were investigated by molecular ecological network and metagenomics analyses. Results showed that the satisfactory nitrogen removal efficiency (>97 %) was achieved in the sulfide−SADN process (S/N molar ratio of 0.88) with 1.5 % salinity. In salinity scenarios, the genus Thiobacillus significantly proliferated and was detected as the dominant sulfur−oxidizing bacteria in the sulfide−SADN system, occupying a relative abundance of 29.4 %. Network analysis further elucidated that 1.5 % salinity had enabled the microbial community to form a more densely clustered network, which intensified the interactions between microorganisms and effectively improved the nitrogen removal performance of the sulfide−SADN. Metagenomics sequencing revealed that the abundance of functional genes encoding for key enzymes involved in SADN, dissimilatory nitrate reduction to ammonium, and nitrification was up−regulated in the 1.5 % salinity scenario compared to that without salinity, stimulating the occurrence of multiple nitrogen transformation pathways. These multi−paths contributed to a robust SADN process (i.e., nitrogen removal efficiency >97 %, effluent nitrogen <2.5 mg N/L). This study deepens our understanding of the effect of salt on the SADN system at the community and functional level, and favors to advance the application of this sustainable bioprocess in saline wastewater treatment.

Seasonal and spatial characteristics of microbial community in intertidal wetlands: Aspect of microbial coupling

The simultaneous reductions of pollution and carbon are widely concerned in sewage treatment. Various microorganisms with synergies are existed in the intertidal wetlands, which have the potential to be the microbial inoculation used in sewage treatment. In this study, the microorganisms in the reed wetlands of the Yellow River Delta in four seasons and at four layers (0–5, 5–10, 10–15, and 15–20 cm) were investigated by high-throughput sequencing technology. The results showed that the majority of functional bacteria were denitrifying bacteria in the spring and summer. Spring was the primary season for hydrogen autotrophic denitrification, whereas summer was the primary season for sulfur autotrophic denitrification. A range of bacteria connected to one another by cable bacteria cooperated to finish the cycle of several elements in the 10–15 cm layer in autumn. A distinctive characteristic of functional bacteria was the anaerobic methane oxidation mediated by NC10 facultative bacteria in the 0–5 cm layer in winter. The results of this study provide an inspiration for utilizing suitable microbial inoculation to realize the simultaneous reductions of pollution and carbon in the future sewage treatment.

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

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

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

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

Amorphous Fe substrate enhances nitrogen and phosphorus removal in sulfur autotrophic process

The autotrophic denitrification of coupled sulfur and natural iron ore can remove nitrogen and phosphorus from wastewater with low C/N ratios. However, the low solubility of crystalline Fe limits its bioavailability and P absorption capacity. This study investigated the effects of amorphous Fe in drinking water treatment residue (DWTR) and crystalline Fe in red mud (RM) on nitrogen and phosphorus removal during sulfur autotrophic processes. Two types of S-Fe cross-linked filler particles with three-dimensional mesh structures were obtained by combining sulfur with the DWTR/RM using the hydrogel encapsulation method. Two fixed-bed reactors, sulfur-DWTR autotrophic denitrification (SDAD) and sulfur-RM autotrophic denitrification (SRAD), were constructed and stably operated for 236 d Under a 5–8-h hydraulic retention time, the average NO3−–N, TN, and phosphate removal rates of SDAD and SRAD were 99.04 %, 96.29 %, 94.03 % (SDAD) and 97.33 %, 69.97 %, 82.26 % (SRAD), respectively. It is important to note that fermentative iron-reducing bacteria, specifically Clostridium_sensu_stricto_1, were present in SDAD at an abundance of 58.17 %, but were absent from SRAD. The presence of these bacteria facilitated the reduction of Fe (III) to Fe (II), which led to the complete denitrification of the S-Fe (II) co-electron donor to produce Fe (III), completing the iron cycle in the system. This study proposes an enhancement method for sulfur autotrophic denitrification using an amorphous Fe substrate, providing a new option for the efficient treatment of low-C/N wastewater.

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.

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.