Autotrophic Denitrification System Research Articles

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 100mg/L) and BAC (0.5, 5 and 10mg/L). After 100 days of operation, the presence of 10mg/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 would broaden the understanding of the environmental hazards posed by PVC MPs and BAC in WWTPs.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Effect of phosphorus on the performance of sulfur autotrophic denitrification

Autotrophic denitrification, recognized as a biological denitrification method, has garnered increasing interest in the field of ecological water environment denitrification technology. This study focuses on the utilization of the sulfur autotrophic denitrification process for the deep denitrification treatment of wastewater. Furthermore, it investigates the impact of phosphorus on nitrogen removal efficiency and bacterial community structure of sulfur autotrophic denitrification. In the experiment, a column containing an autotrophic denitrification filter with a carbon-sulfur filler was constructed. This study aimed to examine the impact of phosphorus on the initiation and denitrification efficiency of the autotrophic denitrification filter column. High-throughput sequencing was used to examine alterations in the structure of the bacterial community. The experimental findings indicated that the absence of phosphorus in the influent considerably extended the initiation period of the autotrophic denitrification filter column. The average Total Inorganic Nitrogen (TIN) removal rate was determined to be 80.26%, while the effluent exhibited an accumulation concentration of NO2−-N ranging from 7 to 11 mg/L. After the addition of phosphate, the removal rate of NO3−-N in the autotrophic denitrification process increased to more than 97%, with no2−-N accumulation. As the influent phosphorus concentration increased, the ability of the sulfur autotrophic denitrification filter column to resist hydraulic impact load improved. At a NO3−-N concentration of 30 mg/L and Hydraulic Retention Time (HRT) = 2 h, the mean NO3−-N removal rate increased from 73.22% to 81.71%. The concentration of phosphorus affected the quantity and variety of primary bacteria in the sulfur autotrophic denitrification system, as their domination changed from Pseudomonas to Ferritrophicum and Thiobacillus, and from a mixed culture to complete autotrophy.

Vertical spatial denitrification performance and microbial community composition in denitrification biofilters coupled with water electrolysis.

In this study, a denitrification biofilter coupled with water electrolysis (DNBF-WE) was developed as a novel heterotrophic-hydrogen autotrophic denitrification system, which could enhance denitrification with limited organic carbon in the secondary effluent. The volumetric denitrification rate of DNBF-WE reached 152.16 g N m-3 d-1 (C/N = 2, I = 60 mA, and HRT = 5 h). Besides, the vertical spatial denitrification of DNBF-WE was explored, with the nitrate removal rate being 49.5%, 16.3%, and 29.3% in the top, middle, and bottom, respectively. The concentration of extracellular polymeric substances (EPSs) was consistent with the denitrification performance vertically. The high-throughput sequencing analysis results revealed that autotrophic denitrification bacteria (e.g. Thauera) gradually enriched along DNBF-WE from top to bottom. The functional gene prediction results illustrated the vertical stratification mechanisms of the denitrification. Both dissimilatory nitrate reduction and denitrification contributed to nitrate removal, and denitrification became more advantageous with an increase in the filter depth. The research on both the performance of DNBF-WE and the characteristics of microbial communities in the vertical zones of the biofilter may lay a foundation for the biofilter denitrification process in practice.

Autotrophic denitrification based on sulfur-iron minerals: advanced wastewater treatment technology with simultaneous nitrogen and phosphorus removal.

Autotrophic denitrification technology has many advantages, including no external carbon source addition, low sludge production, high operating cost efficiency, prevention of secondary sewage pollution, and stable treatment efficiency. At present, the main research on autotrophic denitrification electron donors mainly includes sulfur, iron, and hydrogen. In these autotrophic denitrification systems, pyrite has received attention due to its advantages of easy availability of raw materials, low cost, and pH stability. When pyrite is used as a substrate for autotropic denitrification, sulfide (S2-) and ferrous ion (Fe2+) in the substrate will provide electrons to convert nitrate (NO3-) in sewage first to nitrite (NO2-), then to nitrogen (N2), and finally to discharge the system. At the same time, sulfide (S2-) loses electrons to sulfate (SO42-) and ferrous ion (Fe2+) loses electrons to ferric iron (Fe3+). Phosphates (PO43-) in wastewater are chemically combined with ferric iron (Fe3+) to form ferric phosphate (FePO4) precipitate. This paper aims to provide a detailed and comprehensive overview of the dynamic changes of nitrogen (N), phosphorus (P), and other substances in the process of sulfur autotrophic denitrification using iron sulfide, and to summarize the factors that affect wastewater treatment in the system. This work will provide a relevant research direction and theoretical basis for the field of sulfur autotrophic denitrification, especially for the related experiments of the reaction conversion of various substances in the system.

Microbial community structure characteristics and gene distribution of sulfur-siderite/limestone autotrophic denitrification

Microbial community structure and functional gene of sulfur-siderite/limestone autotrophic denitrification (SSAD/SLAD) systems were comprehensively investigated in bath bioassays by microbiological approaches. High-throughput sequencing analysis showed that there were abundant sulfur autotrophic denitrifying bacteria in both systems, with the dominant genera being Sulfurimonas, Thiomonas, Thiobacillus and Acidithiobacillus. A remarkably higher Thiobacillus abundance in SSAD system was probably because of the presence of Fe2+, while Ignavibacterium, Desulfocapsa and Chlorobium were markedly more active in SLAD system, which could be closely related to its higher concentration of sulfide. Analysis of macrogenome sequencing inferred that the main functional genes implicated in the denitrification process included nrtABC, nar, napAB, nosZ, norBC and nirSK, endowing excellent denitrification efficiency of systems. Meanwhile, the genes involved in sulfate reduction mainly included apr, dsr and sat genes, and sox and dsr participated in sulfur oxidation process. This study provided insights into mechanisms and reliable application of sulfur autotrophic denitrification.

Efficient nitrite accumulation in partial sulfide autotrophic denitrification (PSAD) system: insights of S/N ratio, pH and temperature

ABSTRACT To provide the necessary nitrite for the Anaerobic Ammonium Oxidation (ANAMMOX) process, the effect of nitrite accumulation in the partial sulfide autotrophic denitrification (PSAD) process was investigated using an SBR reactor. The results revealed that the effectiveness of nitrate removal was unsatisfactory when the S/N ratio (mol/mol) fell below 0.6. The optimal conditions for nitrate removal and nitrite accumulation were achieved within the S/N ratio range of 0.7-0.8, resulting in an average Nitrate Removal Efficiency (NRE) of 95.84%±4.89% and a Nitrite Accumulation Rate (NAR) of 75.31%±6.61%, respectively. It was observed that the nitrate reduction rate was three times faster than that of nitrite reduction during a typical cycle test. Furthermore, batch tests were conducted to assess the influence of pH and temperature conditions. In the pH tests, it became evident that the PSAD process performed more effectively in alkaline environment. The highest levels of nitrate removal and nitrite accumulation were achieved at an initial pH of 8.5, resulting in a NRE of 98.30%±1.93% and a NAR of 85.83%±0.47%, respectively. In the temperature tests, the most favourable outcomes for nitrate removal and nitrite accumulation were observed at 22±1 ℃, with a NRE of 100.00% and a NAR of 81.03%±1.64%, respectively. Moreover, a comparative analysis of 16S rRNA sequencing results between the raw sludge and the sulfide-enriched culture sludge sample showed that Proteobacteria (49.51%) remained the dominant phylum, with Thiobacillus (24.72%), Prosthecobacter (2.55%), Brevundimonas (2.31%) and Ignavibacterium (2.04%) emerging as the dominant genera, assuming the good nitrogen performance of the system.

Deciphering the response of heterotrophic, autotrophic and mixotrophic denitrification to enrofloxacin stress: Performance, denitrifying community and antibiotic resistance genes

Antibiotics frequently coexist with nitrate (NO3−-N) in wastewater. This study systematically explored the negative effects of enrofloxacin (ENR) on the microbial performance of autotrophic, heterotrophic and mixotrophic denitrification processes under parallel conditions. The ENR concentration was set at 0, 0.1, 1 and 10 mg/L for four phases in sequence. As the influent ENR concentration increased, the nitrate removal efficiency (NrE) in the pyrite-based autotrophic denitrification (PAD) system declined gradually to 11.08% (minimum) and the expressed narG/H/I and napA/B genes decreased. However, the NrE in the polycaprolactone-based heterotrophic denitrification (PHD) and mixotrophic denitrification systems declined slightly (the lowest NrE: 88.10% and 85.28%, respectively), and the expressed narG/H/I (0–1 mg/L) and napA/B genes increased gradually in the PHD system. The removal pathway of ENR was mainly through adsorption by substrates and/or biofilms rather than through biodegradation. Increasing the concentration of ENR (0–10 mg/L) reduced the relative abundance of functional genera (Syntrophomonas, Lentimicrobiaceae_unclassified, Bacteroidetes_vadinHA17_unclassified, and Clostridium) capable of degrading polymers in the PHD system, but elevated the relative abundance of the dominant genus Thiobacillus in the PAD system. Network analyses showed that Thiobacillus in the PAD system as well as Anaerolineaceae_unclassified and Simplicispira in the PHD system were resistant to ENR stress.

Deciphering responses of sulfur autotrophic denitrification system to the single and joint stress of Zn(II) and dialkyldimethyl ammonium compounds: Performance, microbial community and different fractions of resistance genes

Zn(II) and dialkyldimethyl ammonium compounds (DADMAC) are common heavy metal and disinfectant contaminants in wastewater treatment plants. The response of sulfur autotrophic denitrification (SAD) system under single and joint stress with different concentrations of Zn(II) (1–25 mg/L) and DADMAC (0.2–10 mg/L) was evaluated using sequencing batch fixed-bed biofilm reactors in this study. The joint stress of high concentrations of Zn(II) and DADMAC had stronger inhibitory effects on microbial activity and metabolic function genes compared to the single stress, thus significantly inhibiting the performance of the SAD system. Furthermore, Zn(II) stress contributed to the proliferation of resistance genes (RGs) in water (w-RGs), whereas stress of DADMAC was able to induce the proliferation of intracellular RGs (si-RGs) and extracellular RGs (se-RGs) in sludge. And under the joint stress of Zn(II) and DADAMC, the abundances of si/se-RGs were much higher than that under single stress, with a higher risk of RGs transmission. The α-Helix/(β-sheet + random coil) in tightly bound extracellular polymeric substances correlated positively with si-tnpA-04 and se-tnpA-04. Horizontal gene transfer via mobile genetic elements was the main mode of RGs transmission. This work assessed the risk of DADAMC and Zn(II) in the SAD system and established new insights into the transmission of three-fraction RGs under DADMAC and Zn(II) stress.

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.