Effects of combined antibiotics on nitrification, bacteria and antibiotic resistance genes in activated sludge: Insights from legacy effect of antibiotics. - Related Documents




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791601.0000Effects of combined antibiotics on nitrification, bacteria and antibiotic resistance genes in activated sludge: Insights from legacy effect of antibiotics. The effect of combined antibiotics exposure on nitrogen removal, microbial community assembly and proliferation of antibiotics resistance genes (ARGs) is a hotspot in activated sludge system. However, it is unclear that how the historical antibiotic stress affects the subsequent responses of microbes and ARGs to combined antibiotics. In this study, the effects of combined sulfamethoxazole (SMX) and trimethoprim (TMP) pollution on activated sludge under legacy of SMX or TMP stress with different doses (0.005-30 mg/L) were investigated to clarify antibiotic legacy effects. Nitrification activity was inhibited under higher level of combined exposure but a high total nitrogen removal (∼70%) occurred. Based on the full-scale classification, the legacy effect of past antibiotic stress had a marked effect on community composition of conditionally abundant taxa (CAT) and conditionally rare or abundant taxa (CRAT). Rare taxa (RT) were the keystone taxa in the microbial network, and the responses of hub genera were also affected by the legacy of antibiotic stress. Nitrifying bacteria and genes were inhibited by the antibiotics and aerobic denitrifying bacteria (Pseudomonas, Thaurea and Hydrogenophaga) were enriched under legacy of high dose, as were the key denitrifying genes (napA, nirK and norB). Furthermore, the occurrences and co-selection relationship of 94 ARGs were affected by legacy effect. While, some shared hosts (eg., Citrobacter) and hub ARGs (eg., mdtD, mdtE and acrD) were identified. Overall, antibiotic legacy could affect responses of activated sludge to combined antibiotic and the legacy effect was stronger at higher exposure levels.202337225384
807110.9998Impact of ciprofloxacin and copper combined pollution on activated sludge: Abundant-rare taxa and antibiotic resistance genes. This study aimed to explore the impacts of ciprofloxacin (CIP, 0.05-40 mg/L) and copper (3 mg/L) combined pollution on nitrification, microbial community and antibiotic resistance genes (ARGs) in activated sludge system during stress- and post-effect periods. Higher CIP concentration inhibited nitrification and an average of 50% total nitrogen removal occurred under 40 mg/L of CIP pressure. The stress- and post-effects on bacterial diversity and structure were obviously distinct. Abundant genera were more sensitive to combined pollution than rare genera based on full-scale classification and conditionally rare or abundant taxa were keystone taxa in their interactions. Ammonia oxidation genes were inhibited under high CIP level, but some aerobic denitrifying bacteria (Thauera, Comamonas and Azoarcus) and key genes increased. 96 ARG subtypes were detected with complex positive relationships and their potential hosts (abundant-rare-functional genera) changed in two periods. This study highlights the different stress- and post-effects of combined pollution on activated sludge.202235217161
808220.9998Deciphering the toxic effects of antibiotics on denitrification: Process performance, microbial community and antibiotic resistance genes. The extensive application of antibiotics, and the occurrence and spread of antibiotic resistance genes (ARGs) shade health risks to human and animal. The long-term effects of sulfamethoxazole (SMX) and tetracycline (TC) on denitrification process were evaluated in this study, with the focus on nitrogen removal performance, microbial community and ARGs. Results showed that low-concentration SMX and TC (<0.2 mg L(-1)) initially caused a deterioration in nitrogen removal performance, while higher concentrations (0.4-20 mg L(-1)) of both antibiotics had no further inhibitory influences. The abundances of ARGs in both systems generally increased during the whole period, and most of them had significant correlations with intI1, especially efflux-pump genes. Castellaniella, which was the dominant genus under antibiotic pressure, might be potential resistant bacteria. These findings provide an insight into the toxic effects of different antibiotics on denitrification process, and guides future efforts to control antibiotics pollution in ecosystems.202032250829
808430.9998Metagenomic insights into effects of carbon/nitrogen ratio on microbial community and antibiotic resistance in moving bed biofilm reactor. This study investigated the effects of carbon/nitrogen (C/N) ratio on microbial community in moving bed biofilm reactor (MBBR) using metagenomic analysis, and the dynamic changes of relevant antibiotic resistance genes (ARGs) were also analyzed. The results showed that under low C/N ratio, MBBR exhibited average removal rates of 98.41 % for ammonia nitrogen and 75.79 % for total nitrogen. Metagenomic analysis showed low C/N ratio altered the structure of biofilm and water microbiota, resulting in the detachment of bacteria such as Actinobacteria from biofilm into water. Furthermore, sulfamethazine (SMZ)-resistant bacteria and related ARGs were released into water under low C/N ratio, which lead to the increase of SMZ resistance rate to 90%. Moreover, most dominant genera are potential hosts for both nitrogen cycle related genes and ARGs. Specifically, Nitrosomonas that carried gene sul2 might be released from biofilm into water. These findings implied the risks of antibiotic resistance dissemination in MBBR under low C/N ratio.202438901747
756340.9998Polystyrene microplastics alleviate the effects of sulfamethazine on soil microbial communities at different CO(2) concentrations. Microplastics were reported to adsorb antibiotics and may modify their effects on soil systems. But there has been little research investigating how microplastics may affect the toxicities of antibiotics to microbes under future climate conditions. Here, we used a free-air CO(2) enrichment system to investigate the responses of soil microbes to sulfamethazine (SMZ, 1 mg kg(-1)) in the presence of polystyrene microplastics (PS, 5 mg kg(-1)) at different CO(2) concentrations (ambient at 380 ppm and elevated at 580 ppm). SMZ alone decreased bacterial diversity, negatively affected the bacterial structure and inter-relationships, and enriched the sulfonamide-resistance genes (sul1 and sul2) and class 1 integron (intl1). PS, at both CO(2) conditions, showed little effect on soil bacteria but markedly alleviated SMZ's adverse effects on bacterial diversity, composition and structure, and inhibited sul1 transmission by decreasing the intl1 abundance. Elevated CO(2) had limited modification in SMZ's disadvantages to microbial communities but markedly decreased the sul1 and sul2 abundance. Results indicated that increasing CO(2) concentration or the presence of PS affected the responses of soil microbes to SMZ, providing new insights into the risk prediction of antibiotics under future climate conditions.202133592488
758450.9998Responses of microbial community and antibiotic resistance genes to co-existence of chloramphenicol and salinity. In recent years, the risk from environmental pollution caused by chloramphenicol (CAP) has emerged as a serious concern worldwide, especially for the co-selection of antibiotic resistance microorganisms simultaneously exposed to CAP and salts. In this study, the multistage contact oxidation reactor (MCOR) was employed for the first time to treat the CAP wastewater under the co-existence of CAP (10-80 mg/L) and salinity (0-30 g/L NaCl). The CAP removal efficiency reached 91.7% under the co-existence of 30 mg/L CAP and 10 g/L NaCl in the influent, but it fluctuated around 60% with the increase of CAP concentration and salinity. Trichococcus and Lactococcus were the major contributors to the CAP and salinity shock loads. Furthermore, the elevated CAP and salinity selection pressures inhibited the spread of CAP efflux pump genes, including cmlA, tetC, and floR, and significantly affected the composition and abundance of antibiotic resistance genes (ARGs). As the potential hosts of CAP resistance genes, Acinetobacter, Enterococcus, and unclassified_d_Bacteria developed resistance against high osmotic pressure and antibiotic environment using the efflux pump mechanism. The results also revealed that shifting of potential host bacteria significantly contributed to the change in ARGs. Overall, the co-existence of CAP and salinity promoted the enrichment of core genera Trichococcus and Lactococcus; however, they inhibited the proliferation of ARGs. KEY POINTS: • Trichococcus and Lactococcus were the core bacteria related to CAP biodegradation • Co-existence of CAP and salinity inhibited proliferation of cmlA, tetC, and floR • The microorganism resisted the CAP using the efflux pump mechanism.202236205764
791560.9998Deciphering antibiotic resistance genes and microbial community of anammox consortia under sulfadiazine and chlortetracycline stress. The responses of anammox consortia to typical antibiotics sulfadiazine (SDZ) and chlortetracycline (CTC) were evaluated on the aspects of general performance, microbial activity, diversity and abundance of antibiotic resistance genes (ARGs), and microbial host of ARGs in anammox system. Results showed the anammox consortia had a stable performance and great resistance to 10 mg/L of SDZ, while 1 mg/L of CTC induced an unrecoverable inhibitory influence on nitrogen removal performance and anammox activity without any special treatment. The absolute abundances of anammox functional genes (nirS, hzsA and hdh) were stimulated by the acclimation to SDZ stress, however, they were much lower than the initial levels under CTC stress. In anammox consortia, ARGs comprised 18 types (94 subtypes) derived from over 20 genera. Strikingly, the anammox bacteria (AnAOB) "Ca. Brocadia" occupied 46.81% of the SDZ resistance genes (sul1 and sul2) and 38.63% of CTC resistance genes (tetX, tetG and rpsJ), and thus were identified as the dominant antibiotic resistance bacteria (ARB). Therefore, harboring the corresponding ARGs by AnAOB could be the primary protective mechanism to interpret the resistance of anammox consortia to antibiotics stress. Meanwhile, co-occurring of ARGs in anammox consortia suggested the synergistic cooperation of different ARGs could be an essential strategy to alleviate the SDZ and CTC stress. The present study proposed a new interpretation of possible mechanism that cause antibiotic resistance of anammox consortia.202235259594
797870.9997Metagenomic insight into the enrichment of antibiotic resistance genes in activated sludge upon exposure to nanoplastics. Activated sludge is an important reservoir for the co-occurring emerging contaminants including nanoplastics (NPs) and antibiotic resistance genes (ARGs). However, the impacts and potential mechanisms of NPs on the fate of ARGs in activated sludge are not fully understood. Herein, we used metagenomic approach to investigate the responses of ARGs, host bacteria, mobile genetic elements (MGEs), and functional genes to polystyrene (PS) NPs at environmentally relevant (0.5 mg/L) and high stress concentrations (50 mg/L) in activated sludge. The results showed that 0.5 and 50 mg/L PS NPs increased the relative abundance of ARGs in the activated sludge by 58.68% and 46.52%, respectively (p < 0.05). Host tracking analysis elucidated that the hosts of ARGs were significantly enriched by PS NPs (p < 0.05), with Proteobacteria being the predominant host bacteria. Additionally, the occurrence of new ARGs hosts and the enrichment of MGEs and functional genes (i.e., genes related to SOS response, cell membrane permeability, and secretion system, etc.) indicated that PS NPs promoted horizontal gene transfer (HGT) of ARGs. Finally, path modeling analysis revealed that the proliferation of ARGs caused by PS NPs was primarily attributed to the enhancement of HGT and the enrichment of host bacteria. Our findings contribute to a comprehensive understanding of the spread risk of ARGs in activated sludge under NPs pollution.202439510298
791380.9997Response of the partial denitrification coupled with anaerobic ammonia oxidation system to disinfectant residues stress. The extensive use of disinfectants, especially NaClO, has resulted in chlorine disinfectant residues entering and impairing the biological treatment system. This study combined with long-term stress and transient shock of chlorine residues to comprehensively evaluate the variations of nitrogen removal performance, microbial community and antibiotic resistance genes composition in the PD/A system. The results showed that low concentration NaClO had no obvious harm to the system, but high concentration (>1 mg/L) NaClO would destroy the nitrogen removal performance of PD/A system. Interestingly, microorganisms in biofilm were more resistant to chlorine residues than that in sludge. Anaerobic ammonia oxidizing bacteria suffered more harm than denitrifying microorganisms, and chlorine residues mainly inhibited the process of converting N(2)H(4) to N(2) in anammox reaction. In addition, this study found that sludge showed a more significant increase in ARGs abundance and risk than biofilm. Moreover, risk assessments indicated that chlorine residues increased the risk of ARGs in PD/A systems.202540010223
808690.9997Biofilm enhanced the mitigations of antibiotics and resistome in sulfadiazine and trimethoprim co-contaminated soils. Reducing antibiotic levels in soil ecosystems is vital to curb the dissemination of antimicrobial resistance genes (ARGs) and mitigate global health threats. However, gaps persist in understanding how antibiotic resistome can be suppressed during antibiotic degradation. Herein, we investigate the efficacy of a biochar biofilm incorporating antibiotics-degrading bacterial strain (Arthrobacter sp. D2) to mitigate antibiotic resistome in non-manured and manure-amended soils with sulfadiazine (SDZ) and trimethoprim (TMP) contamination. Results show that biofilm enhanced SDZ degradation by 83.0% within three days and increased TMP attenuation by 55.4% over 60 days in non-manured soils. In the non-manured black soil, the relative abundance of ARGs increased initially after biofilm inoculation. However, by day 30, it decreased by 20.5% compared to the controls. Moreover, after 7 days, biofilm reduced TMP by 38.5% in manured soils and decreased the total ARG abundance by 19.0%. Thus, while SDZ degradation did not increase sulfonamide resistance genes, TMP dissipation led to a proliferation of insertion sequences and related TMP resistance genes. This study underscores the importance of antibiotic degradation in reducing related ARGs while cautioning against the potential proliferation and various ARGs transfer by resistant microorganisms.202439255667
7560100.9997The effect of bacterial functional characteristics on the spread of antibiotic resistance genes in Expanded Granular Sludge Bed reactor treating the antibiotic wastewater. To explore the fate and spreading mechanism of antibiotics resistance genes (ARGs) in antibiotics wastewater system, a laboratory-scale (1.47 L) Expanded Granular Sludge Bed (EGSB) bioreactor was implemented. The operating parameters temperature (T) and hydraulic retention time (HRT) were mainly considered. This result showed the removal of ARGs and COD was asynchronous, and the recovery speed of ARGs removal was slower than that COD removal. The decreasing T was attributed to the high growth rate of ARGs host bacteria, while the shortened HRT could promote the horizontal and vertical gene transfer of ARGs in the sludge. The analysis result of potential bacterial host showed more than half of the potential host bacteria carried 2 or more ARGs and suggested an indirect mechanism of co-selection of multiple ARGs. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) was used to investigate the functional characteristics of bacterial community. This result showed the bacterial functional genes contributed 40.41% to the abundance change of ARGs in the sludge, which was higher that of bacterial community. And the function genes of "aromatic hydrocarbon degradation", "Replication, recombination and repair proteins" and "Flagellar assembly" were mainly correlated with the transfer of ARGs in the sludge. This study further revealed the mechanism of ARGs spread in the EGSB system, which would provide new ideas for the development of ARGs reduction technology.202134488144
7564110.9997Formation, characteristics and microbial community of aerobic granular sludge in the presence of sulfadiazine at environmentally relevant concentrations. The growing occurrence of antibiotics in water environment is causing increasing concern. To investigate the impact of frequently detected sulfadiazine on the formation of aerobic granular sludge, four sequencing batch reactors (SBRs) were set up with different environmentally relevant concentrations of sulfadiazine. Results showed that sulfadiazine pressure could lead to larger and more compact sludge particles and cause slight effect on reactor performance. Presence of sulfadiazine apparently increased the extracellular polymeric substances (EPS) secretion of microorganisms. Quantitative polymerase chain reaction (qPCR) showed that the abundances of sulfanilamide resistance genes in sludge increased with addition of sulfadiazine significantly. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) was used to predict functional genes, results showed that sulfadiazine led to an increase of specific functional genes. Thereby, it concluded that microorganisms could change the community structure by acclimating of functional bacteria and antibiotic resistance species to adapt to the antibiotic stress.201829197771
8592120.9997Effects of persulfate treatment on the fates of antibiotic resistance genes in waste activated sludge fermentation process and the underlying mechanism. The occurrence of antibiotic resistance genes (ARGs) in waste activated sludge (WAS) fermentation was investigated with persulfate (PS)-based treatment. ARGs affiliated with multidrug (mexP), macrolide (bla(OXA-129)), tetracycline (tetB), sulfonamide (sul1), and vancomycin (vanRG) types were significantly decreased by PS/Fe treatment. Mechanistic investigations revealed that PS/Fe possessed oxidating potential and exhibited devastating effects on WAS fermentation. First, PS/Fe promoted cell structure damage, which facilitated ARGs release from potential hosts. A co-occurrence network analysis indicated that Fe/PS suppressed the proliferation of potential host bacteria. In addition, the PS/Fe treatment induced the decreased abundance of certain functional genes involved in pathways associated with ARGs dissemination. Finally, variation partitioning analysis demonstrated that the microbial community structure exhibited more vital effects on ARGs fates than physicochemical factors (i.e., pH and ORP) and gene expression (i.e., two-component system). This work provided a deeper understanding of the critical factors used to determine ARGs fates during WAS fermentation.202234864181
8041130.9997Insights into the microalgae-bacteria consortia treating swine wastewater: Symbiotic mechanism and resistance genes analysis. This study investigated the effects of microalgae-bacteria consortia (MBC) (Chlorella pyrenoidosa-activated sludge (AS)) treating swine wastewater with low C/N ratios. After co-culture, the removal rates of NH(4)(+)-N and PO(4)(3-)-P increased by 53.84% and 43.52%. Furthermore, the sulfamethoxazole (SMX) degradation rates in MBC were slightly higher than in the activated sludge process. Interestingly, the absolute abundance of antibiotic resistance genes (ARGs) in effluent from MBC is relatively less than in the AS process. C. pyrenoidosa has a negative zeta potential that allows bacteria to adhere to its surface. The concentrations of carbohydrates and proteins in extracellular polymeric substance (EPS) of MBC dramatically increased compared with the AS process. At the phylum level, Proteobacteria, Bacteroidota, and Cyanobacteria were the main bacteria, while Ascomycota and Basidiomycota were the primary fungi in MBC. Overall, those findings lead to a better understanding of the swine wastewater containing antibiotic treatment by MBC.202235217162
7562140.9997Nanoparticles and antibiotics stress proliferated antibiotic resistance genes in microalgae-bacteria symbiotic systems. The comprehensive effect of exogenous pollutants on the dispersal and abundance of antibiotic-resistance genes (ARGs) in the phycosphere, bacterial community and algae-bacteria interaction remains poorly understood. We investigated community structure and abundance of ARGs in free-living (FL) and particle-attached (PA) bacteria in the phycosphere under nanoparticles (silver nanoparticles (AgNPs) and hematite nanoparticles (HemNPs)) and antibiotics (tetracycline and sulfadiazine) stress using high-throughput sequencing and real-time quantitative PCR. Meanwhile, the intrinsic connection of algae-bacteria interaction was explored by transcriptome and metabolome. The results showed that the relative abundance of sulfonamide and tetracycline ARGs in PA and FL bacteria increased 103-129 % and 112-134 %, respectively, under combined stress of nanoparticles and antibiotics. Antibiotics have a greater effect on ARGs than nanoparticles at environmentally relevant concentrations. Proteobacteria, Firmicutes, and Bacteroidetes, as the primary potential hosts of ARGs, were the dominant phyla. Lifestyle, i.e., PA and FL, significantly determined the abundance of ARGs and bacterial communities. Moreover, algae can provide bacteria with nutrients (carbohydrates and amino acids), and can also produce antibacterial substances (fatty acids). This algal-bacterial interaction may indirectly affect the distribution and abundance of ARGs. These findings provide new insights into the distribution and dispersal of ARGs in microalgae-bacteria symbiotic systems.202336283215
7565150.9997Microalgae Enhances the Adaptability of Epiphytic Bacteria to Sulfamethoxazole Stress and Proliferation of Antibiotic Resistance Genes Mediated by Integron. The transmission of ARGs in the microalgae-associated epiphytic bacteria remains unclear under antibiotic exposure, apart from altering the microbial community structure. In this study, Chlorella vulgaris cocultured with bacteria screened from surface water was examined to explore the spread of ARGs in the presence of sulfamethoxazole (SMX). The extracellular polymers released by Chlorella vulgaris could reduce antibiotic-induced collateral damage to bacteria, thus increasing the diversity of the microalgae-associated epiphytic bacteria. The abundances of sul1 and intI1 in the phycosphere at 1 mg/L SMX dose increased by 290 and 28 times, respectively. Metagenomic sequencing further confirmed that SMX bioaccumulation stimulated the horizontal transfer of sul1 mediated by intI1 in the microalgae-associated epiphytic bacteria, while reactive oxygen species (ROS)-mediated oxidative stress induced the SOS response and thus enhanced the transformation of sul1 in the J group. This is the first study to verify that microalgae protect bacteria from antibiotic damage and hinder the spread of ARGs mediated by SOS response, while the transfer of ARGs mediated by integron is promoted due to the bioaccumulation of SMX in the phycosphere. The results contribute to present comprehensive understanding of the risk of ARG proliferation by the presence of emerging contaminants residues in river.202439417646
7566160.9997Antibiotic sulfadiazine degradation by persulfate oxidation: Intermediates dependence of ecotoxicity and the induction of antibiotic resistance genes. To preserve the water resources, this study has analyzed the ecotoxicity and antibiotic resistance genes (ARGs) induction capacity of sulfadiazine degradation intermediates resulting from persulfate activation oxidation enhanced by ultraviolet, ultrasound and microwave. The five degradation pathways caused by the contribution discrepancy of electron transfer and singlet oxygen ((1)O(2)) and variations in the ecotoxicity of different degradation products were analyzed. Microcosm experiment exhibited that the microbial community in actual water changed significantly with SDZ and degradation intermediates, in which the dominant genera were Aeromonas, Cupriavidus, Elizabethkingia and Achromobacter. Except for the selective pressure on bacteria, the degradation intermediates also exert a certain degree or even stronger induction on sulfonamide ARGs (sul4, sul1 and sul2) than SDZ. Furthermore, the potential hosts for sulfonamide ARGs were revealed by network analysis. These results provide a better understanding of antibiotics degradation mechanism and ARGs occurrence, which is useful for controlling the spread of ARGs.202336372382
7959170.9997Evolution of microbial community and antibiotic resistance genes in anammox process stressed by oxytetracycline and copper. The individual and combined impacts of copper ion (Cu(2+)) and oxytetracycline (OTC) on anaerobic ammonium oxidation (anammox) performance and its self-recovery process were examined. Experimental results showed that the anammox performance and activity of anammox bacteria were inhibited by 1.0 mg L(-1) OTC, Cu(2+) and OTC + Cu(2+), and both single and combined inhibitions were reversible. The abundance of functional genes and parts of antibiotic resistance genes (ARGs) were positively related to the dominant bacterium Ca. Kuenenia, implying that the recovery of the performance was associated with the progressive induction of potentially resistant species after inhibition. The above outcomes illustrated that anammox bacteria were stressed by metals and antibiotics, but they still could remove nitrogen at a rate higher than 20.6 ± 0.8 kg N m(-3) d(-1), providing guidance for engineering applications of anammox processes.202132949830
7902180.9997Determination of the lower limits of antibiotic biodegradation and the fate of antibiotic resistant genes in activated sludge: Both nitrifying bacteria and heterotrophic bacteria matter. Antibiotics can be biodegraded in activated sludge via co-metabolism and metabolism. In this study, we investigated the biodegradation pathways of sulfamethoxazole (SMX) and antibiotic resistant genes' (ARGs) fate in different autotrophic and heterotrophic microorganisms, by employing aerobic sludge, mixed sludge, and nitrifying sludge. A threshold concentration of SMX activating the degradation pathways in the initial stage of antibiotics degradation was found and proved in different activated sludge systems. Heterotrophic bacteria played an important role in SMX biodegradation. However, ammonia-oxidizing bacteria (AOB) had a faster metabolic rate, which was about 15 times higher than heterotrophic bacteria, contributing much to SMX removal via co-metabolism. As SMX concentration increases, the amoA gene and AOB relative abundance decreased in aerobic sludge due to the enrichment of functional heterotrophic bacteria, while it increased in nitrifying sludge. Microbial community analysis showed that functional bacteria which possess the capacity of SMX removal and antibiotic resistance were selected by SMX pressure. Potential ARGs hosts could increase their resistance to the biotoxicity of SMX and maintain system performance. These findings are of practical significance to guide antibiotic biodegradation and ARGs control in wastewater treatment plants.202234799165
8044190.9997Effect of tetracycline on nitrogen removal in Moving Bed Biofilm Reactor (MBBR) System. The effect of tetracycline (TC) on nitrogen removal in wastewater treatment plants has become a new problem. This study investigated the effects of TC on nitrogen removal using a Moving Bed Biofilm Reactor system. The results showed that there was no significant effect on nitrogen removal performance when the concentration of TC was 5 mg/L, and that the total nitrogen (TN) removal efficiency could reach 75-77%. However, when the concentration of TC increased to 10 mg/L, the denitrification performance was affected and the TN removal efficiency decreased to 58%. The abundance of denitrifying bacteria such as those in the genus Thauera decreased, and TC-resistant bacteria gradually became dominant. At a TC concentration of 10 mg/L, there were also increases and decreases, respectively, in the abundance of resistance and denitrification functional genes. The inhibitory effect of TC on denitrification was achieved mainly by the inhibition of nitrite-reducing bacteria.202235007308