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853100.9938Biotransformation mechanism of Vibrio diabolicus to sulfamethoxazole at transcriptional level. Sulfamethoxazole (SMX) has attracted much attention due to its high probability of detection in the environment. Marine bacteria Vibrio diabolicus strain L2-2 has been proven to be able to transform SMX. In this study, the potential resistance and biotransformation mechanism of strain L2-2 to SMX, and key genes responses to SMX at environmental concentrations were researched. KEGG pathways were enriched by down-regulated genes including degradation of L-Leucine, L-Isoleucine, and fatty acid metabolism. Resistance mechanism could be concluded as the enhancement of membrane transport, antioxidation, response regulator, repair proteins, and ribosome protection. Biotransformation genes might involve in arylamine N-acetyltransferases (nat), cytochrome c553 (cyc-553) and acyl-CoA synthetase (acs). At the environmental concentration of SMX (0.1-10 μg/L), nat was not be activated, which meant the acetylation of SMX might not occur in the environment; however, cyc-553 was up-regulated under SMX stress of 1 μg/L, which indicated the hydroxylation of SMX could occur in the environment. Besides, the membrane transport and antioxidation of strain L2-2 could be activated under SMX stress of 10 μg/L. The results provided a better understanding of resistance and biotransformation of bacteria to SMX and would support related researches about the impacts of environmental antibiotics.202133429311
788610.9935Resistance of anammox granular sludge to copper nanoparticles and oxytetracycline and restoration of performance. Nanoparticles and antibiotics, the two most frequently detected emerging pollutants from different wastewater sources, are eventually discharged into wastewater treatment plants. In this study, the widely used materials CuNPs and oxytetracycline (OTC) were selected as target pollutants to investigate their joint effects on anaerobic ammonium oxidation (anammox). The results indicated that the environmental concentration slightly inhibited the performance of the reactors, while the performance rapidly deteriorated within a week under high-level combined shocks (5.0 mg L(-1) CuNPs and 2.0 mg L(-1) OTC). After the second shock (2.5 mg L(-1) CuNPs and 2.0 mg L(-1) OTC), the resistance of anammox bacteria was enhanced, with an elevated relative abundance of Candidatus Kuenenia and absolute abundance of hzsA, nirS, and hdh. Moreover, the extracellular polymeric substance (EPS) content and specific anammox activity (SAA) showed corresponding changes. Improved sludge resistance was observed with increasing CuNP and OTC doses, which accelerated the recovery of performance.202032244076
787620.9935Sulfamethoxazole impact on pollutant removal and microbial community of aerobic granular sludge with filamentous bacteria. In this study, sulfamethoxazole (SMX) was employed to investigate its impact on the process of aerobic granule sludge with filamentous bacteria (FAGS). FAGS has shown great tolerance ability. FAGS in a continuous flow reactor (CFR) could keep stable with 2 μg/L of SMX addition during long-term operation. The NH(4)(+), chemical oxygen demand (COD), and SMX removal efficiencies kept higher than 80%, 85%, and 80%, respectively. Both adsorption and biodegradation play important roles in SMX removal for FAGS. The extracellular polymeric substances (EPS) might play important role in SMX removal and FAGS tolerance to SMX. The EPS content increased from 157.84 mg/g VSS to 328.22 mg/g VSS with SMX addition. SMX has slightly affected on microorganism community. A high abundance of Rhodobacter, Gemmobacter, and Sphaerotilus of FAGS may positively correlate to SMX. The SMX addition has led to the increase in the abundance of the four sulfonamide resistance genes in FAGS.202336871701
854130.9934Insights into the response of anammox process to oxytetracycline: Impacts of static magnetic field. The long-term effects of oxytetracycline (OTC) with a high concentration on the anaerobic ammonium oxidation (Anammox) process were evaluated, and the role of static magnetic field (SMF) was further explored. The stress of OTC at 50 mg/L had little effect on the nitrogen removal of anammox process at the first 16 days. With the continuous addition of OTC and the increase of nitrogen loading, the OTC inhibited the nitrogen removal and anammox activity severely. During the 32 days of recovery period without OTC addition, the nitrogen removal was further deteriorated, indicating the inhibition of OTC on anammox activity was irreversible and persistent. The application of SMF alleviated the inhibition of OTC on anammox to some extent, and the specific anammox activity was enhanced by 47.1% compared to the system without SMF during the OTC stress stage. Antibiotic efflux was the major resistance mechanism in the anammox process, and tetA, tetG and rpsJ were the main functional antibiotic resistance genes. The addition of OTC weakened the metabolic interactions between the anammox bacteria and the symbiotic bacteria involved in the metabolism of cofactors and secondary metabolites, leading to the poor anammox activity. The adaptability of microbes to the OTC stress was improved by the application of SMF, which can enhance the metabolic pathways related to bacterial growth and resistance to environmental stress.202337586490
788540.9933Susceptibility, resistance and resilience of anammox biomass to nanoscale copper stress. The increasing use of engineered nanoparticles (NPs) poses an emerging challenge to biological wastewater treatment. The long-term impact of CuNPs on anaerobic ammonium oxidation (anammox) process was firstly investigated in this study. The nitrogen removal capacity of anammox reactor was nearly deprived within 30days under the stress of 5.0mgL(-1) CuNPs and the relative abundance of anammox bacteria (Ca. Kuenenia) was decreased from 29.59% to 17.53%. Meanwhile, copper resistance genes associated with the Cus, Cop and Pco systems were enriched to eliminate excess intracellular copper. After the withdrawal of CuNPs from the influent, the nitrogen removal capacity of anammox biomass recovered completely within 70days. Overall, anammox biomass showed susceptibility, resistance and resilience to the stress of CuNPs. Therefore, the potential impacts of ENPs on anammox-based processes should be of great concern.201728550773
783650.9933Efficient Degradation of Intracellular Antibiotic Resistance Genes by Photosensitized Erythrosine-Produced (1)O(2). Intracellular antibiotic resistance genes (iARGs) constitute the important part of wastewater ARGs and need to be efficiently removed. However, due to the dual protection of intracellular DNA by bacterial membranes and the cytoplasm, present disinfection technologies are largely inefficient in iARG degradation. Herein, we for the first time found that erythrosine (ERY, an edible dye) could efficiently degrade iARGs by producing abundant (1)O(2) under visible light. Seven log antibiotic-resistant bacteria were inactivated within only 1.5 min, and 6 log iARGs were completely degraded within 40 min by photosensitized ERY (5.0 mg/L). A linear relationship was established between ARG degradation rate constants and (1)O(2) concentrations in the ERY photosensitizing system. Surprisingly, a 3.2-fold faster degradation of iARGs than extracellular ARGs was observed, which was attributed to the unique indirect oxidation of iARGs induced by (1)O(2). Furthermore, ERY photosensitizing was effective for iARG degradation in real wastewater and other photosensitizers (including Rose Bengal and Phloxine B) of high (1)O(2) yields could also achieve efficient iARG degradation. The findings increase our knowledge of the iARG degradation preference by (1)O(2) and provide a new strategy of developing technologies with high (1)O(2) yield, like ERY photosensitizing, for efficient iARG removal.202337531556
791860.9932Robustness of the partial nitrification-anammox system exposing to triclosan wastewater: Stress relieved by extracellular polymeric substances and resistance genes. The partial nitrification-anammox (PN/A) process is a promising method for the treatment of municipal wastewater. It is necessary to clarify the responses of PN/A system to antimicrobial agent triclosan (TCS) widely existed in the influent of wastewater treatment plants. In this study, it was found that PN/A system was robust to cope with 0.5 mg/L TCS. Specifically, the control reactor reached 80% total nitrogen removal efficiency (TNRE) on day 107, while the reactor feeding with 0.5 mg/L TCS reached the same TNRE on day 84. The results of the activity test, high-throughput sequencing and DNA-based stable isotope probing showed that 0.5 mg/L TCS did not impede the performance of ammonia oxidizing archaea, ammonia oxidizing bacteria (Nitrosomonas) and anammox bacteria (Candidatus Brocadia and Ca. Kuenenia), but significant inhibited the nitrite oxidizing bacteria (Nitrospira and Ca. Nitrotoga) and denitrifying bacteria. The influent TCS led to the increase of EPS content and enrichment of four resistance genes (RGs) (intI1, sul1, mexB, and tnpA), which might be two principal mechanisms by which PN/A can resist TCS. In addition, functional bacteria carrying multiple RGs also contributed to the maintenance of PN/A system function. These findings improved the understandings of antimicrobial effects on the PN/A system.202234954146
788170.9932Bacterial community shift and antibiotics resistant genes analysis in response to biodegradation of oxytetracycline in dual graphene modified bioelectrode microbial fuel cell. This study explored the biodegradation mechanisms of oxytetracycline (OTC/O) and electrochemical characteristics from the perspective of bacterial community shift and OTC resistance genes in dual graphene modified bioelectrode microbial fuel cell (O-D-GM-BE MFC). In phylum level, Proteobacteria was accounted to 95.04% in O-GM-BA, Proteobacteria and Bacteroidetes were accounted to 59.13% and 20.52% in O-GM-BC, which were beneficial for extracellular electron transport (EET) process and OTC biodegradation. In genus level, the most dominant bacteria in O-GM-BA were Salmonella and Trabulsiella, accounting up to 83.04%, moreover, representative exoelectrogens (Geobacter) were enriched, which contributed to OTC biodegradation and electrochemical performances; abundant degrading bacteria (Moheibacter, Comamonas, Pseudomonas, Dechloromonas, Nitrospira, Methylomicrobium, Pseudorhodoferax, Thiobacillus, Mycobacterium) were enriched in O-GM-BC, which contributed to the maximum removal efficiency of OTC; coding resistance genes of efflux pump, ribosome protective protein and modifying or passivating were all found in O-GM-BE, and this explained the OTC removal mechanisms from gene level.201930640017
775180.9931A novel hypothermic strain, Pseudomonas reactans WL20-3 with high nitrate removal from actual sewage, and its synergistic resistance mechanism for efficient nitrate removal at 4 °C. Nitrate can be well removed by bacteria at 25-30 °C. However, nitrate removal almost ceases at temperatures lower than 5 °C. In this study, a novel hypothermic strain, Pseudomonas reactans WL20-3 exhibited an excellent aerobic nitrate removal ability at 4 °C. It had high capability for the removal of nitrate, total dissolved nitrogen (TDN), and dissolved organic carbon (DOC) at 4 °C, achieving removal efficiencies of 100%, 87.91%, and 97.48%, respectively. The transcriptome analysis revealed all genes involved in the nitrate removal pathway were significantly up-regulated. Additionally, the up-regulation of ABC transporter genes and down-regulation of respiratory chain genes cooperated with the nitrate metabolism pathway to resist low-temperature stress. In actual sewage, inoculated with WL20-3, the nitrate removal efficiency was found to be 70.70%. Overall, these findings demonstrated the impressive capacity of the novel strain WL20-3 to remove nitrate and provided novel insights into the synergistic resistance mechanism of WL20-3 at low temperature.202337369315
788390.9931Anammox biofilm system under the stress of Hg(II): Nitrogen removal performance, microbial community dynamic and resistance genes expression. The existence of heavy metals in wastewater has obtained more attention due to its high toxicity and non-degradability. In this study, we investigated the changes of anaerobic ammonium oxidation (Anammox) system under long-term invasion of Hg(Ⅱ). The results indicated that the total nitrogen removal efficiency (TNRE) dropped to around 55 % as Hg(Ⅱ) concentration went up to 20 mg L(-1). But the functional bacteria rapidly developed some resistant abilities and maintained a stable TNRE of 65 % till the end of test. The maximum relative expression fold change of merA, merB, merD and merR were 468.8476, 23.7383, 5.0321 and 15.2514 times, respectively. The high positive correlation between the expression abundance of metal resistance genes and the concentrations of Hg(Ⅱ) revealed the resistant mechanisms of microorganisms to heavy metals. Moreover, the protective strategy based on extracellular polymeric substances also contributed to the stability of Anammox system.202032315795
8530100.9931Intrinsic chlorine resistance of bacteria modulated by glutaminyl-tRNA biosynthesis in drinking water supply systems. The existence of chlorine-resistant bacteria (CRB) in drinking water supply systems (DWSSs) results in significant challenges to the biological security of drinking water. However, little is known about the intrinsic chlorine-resistant molecular metabolic mechanism of bacteria in DWSSs. This research explored the microbial interactions and the key metabolic pathways that modulate the chlorine resistance of bacteria in full-scale chloraminated DWSSs. The dominant CRB, including Bdellovibrio, Bradyrhizobium, Peredibacter, Sphingomonas, and Hydrogenophaga, strongly interacted with each other to maintain basic metabolism. A total of 4.21% of the bacterial metabolic pathways were key and specific to chlorine-resistant bacteria. Glutaminyl-tRNA biosynthesis was the dominant metabolic pathway of CRB in the target DWSSs. After chloramine disinfection, the relative abundance of glutamate-tRNA ligase (GlnRS) and the related orthologous genes increased by 10.11% and 14.58%, respectively. The inactivation rate of the GlnRS overexpression strain (81.40%) was lower than that of the wild-type strain (90.11%) after exposure to chloramine. Meanwhile, the growth rate of the GlnRS overexpression strain was higher than that of the wild-type strain. Glutaminyl-tRNA biosynthesis can enhance chlorine resistance in DWSSs.202236084827
7914110.9930Response of partial nitrification sludge to the single and combined stress of CuO nanoparticles and sulfamethoxazole antibiotic on microbial activity, community and resistance genes. Considering the inevitable release of antibiotics and nanoparticles (NPs) into the nitrogen containing wastewater, the combined impact of CuO NPs and sulfamethoxazole (SMX) antibiotic on partial nitrification (PN) process was investigated in four identical reactors. Results showed that the bioactivity of the aerobic ammonia-oxidizing bacteria (AOB) decreased by half after they were exposed to the combination of CuO NPs and SMX for short-term; however, there was no obvious variation in the bioactivity of AOB when they were exposed to either CuO NPs or SMX. During long-term exposure, the ammonia removal efficiency (ARE) of CuO NPs improved whereas that of SMX decreased, while the combination of CuO NPs and SMX significantly decreased ARE from 62.9% (in control) to 38.2% and had an unsatisfactory self-recovery performance. The combination of CuO NPs and SMX significantly changed the composition of microbial community, decreased the abundance of AOB, and significantly suppressed PN process. Reegarding the resistance genes, the CuO NPs-SMX combination did not improve the expression of copA, cusA, sul1 and sul2; however, it significantly induced the expression of sul3 and sulA.202032050397
7971120.9930Abiotic mechanism changing tetracycline resistance in root mucus layer of floating plant: The role of antibiotic-exudate complexation. Antibiotic contamination and antibiotic resistance have caused growing concerns in different aquatic environments. This work investigated the complexation between tetracycline chloride (TCH) and the molecular weight (MW)-fractionated root exudates - the key abiotic mechanism impacting antibiotic fate and antibiotic resistance in rhizosphere. Results show that the affinity of TCH to the high MW exudates (≥10 kDa) facilitated the TCH deposition on roots and meanwhile reinforced the expression of certain tetracycline resistance genes (i.e. tetA) and the growth of tetracycline resistant bacteria. The interaction between TCH and the lower MW exudates (<10 kDa) completely inhibited the bacteria growth even below the minimum inhibitory concentration of TCH. In microcosms, the abiotic interaction between TCH and root exudates made effects along with biotic processes. Persistent TCH stimulation (≥50 µg/L, 7 d) induced the change of tet gene abundance and bacteria phyla composition though the mediation of root exudates made the rhizosphere less sensitive to the TCH stress. Summarily, the affinity of antibiotics to root exudates varied with MWs, which was closely related to (i) the antibiotic fate in the root mucus layer, (ii) the bacteria inhibition capacity of antibiotics, and (iii) the antibiotic resistance and bacterial community.202133813291
7884130.9930Underlying the inhibition mechanisms of sulfate and lincomycin on long-term anaerobic digestion: Microbial response and antibiotic resistance genes distribution. This study evaluated the resilience of a long-term anaerobic treatment system exposed to sulfate, lincomycin (LCM) and their combined stress. LCM was found to impede anaerobic propionate degradation, while sulfate for restraining methanogenic acetate utilization. The combined stress, with influent LCM of 200 mg/L and sulfate of 1404 mg/L, revealed severer inhibition on anaerobic digestion than individual inhibition, leading to 73.9 % and 38.5 % decrease in methane production and sulfate removal, respectively. Suppression on propionate-oxidizing bacteria like unclassified_f__Anaerolineae and unclassified_f__Syntrophaceae further demonstrated LCM's inhibitory effect on propionate degradation. Besides, the down-regulation of genes encoding dissimilatory sulfate reduction enzymes caused by LCM triggered great inhibition on sulfate reduction. A notable increase in ARGs was detected under sulfate-stressed condition, owing to its obvious enrichment of tetracycline-resistant genes. Genera including unclassified_f__Syntrophaceae, unclassified_f__Geobacteraceae and unclassified_f__Anaerolineaceae were identified as dominant host of ARGs and enriched by sulfate addition. Overall, these results could provide the theoretical basis for further enhancement on anaerobic digestion of pharmaceutical wastewater containing sulfate and lincomycin.202438185146
7915140.9930Deciphering 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
7911150.9929Biochar induced inhibitory effects on intracellular and extracellular antibiotic resistance genes in anaerobic digestion of swine manure. Distribution of intracellular (iARGs) and extracellular ARGs (eARGs) in manure anaerobic digestion (AD) process coupled with two types of biochar (BC and BP) were investigated. And the effects of biochar on the conjugation transfer of ARGs were explored by deciphering the interaction of biochar with bacterial stress responses, physiological metabolism and antibiotic resistances. Results showed that AD process could effectively remove all the detected eARGs with efficiency of 47.4-98.2%. The modified biochar (BP) with larger specific surface area (SSA) was propitious to decrease the absolute copy number of extracellular resistance genes. AD process could effectively remove iARGs by inhibiting the growth of host bacteria. The results of structural equation models (SEM) indicated that biochar put indirect influences on the fate of ARGs (λ = -0.23, P > 0.05). Analysis on oxidative stress levels, antioxidant capacity, DNA damage-induced response (SOS) response and energy generation process demonstrated that biochar induced the oxidative stress response of microorganisms and enhanced the antioxidant capacity of bacteria. The elevated antioxidant capacity negatively affected SOS response, amplified cell membrane damage and further weakened the energy generation process, resulted in the inhibition of horizontal transfer of ARGs.202235609652
7860160.9929Enhanced removal of antibiotic-resistant bacteria and resistance genes by three-dimensional electrochemical process using MgFe(2)O(4)-loaded biochar as both particle electrode and catalyst for peroxymonosulfate activation. In this study, MgFe(2)O(4)-loaded biochar (MFBC) was used as a three-dimensional particle electrode to active peroxymonosulfate (EC/MFBC/PMS) for the removal of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). The results demonstrated that, under the conditions of 1.0 mM PMS concentration, 0.4 g/L material dosage, 5 V voltage intensity, and MFBC preparation temperature of 600 °C, the EC/MFBC600/PMS system achieved complete inactivation of E. coli DH5α within 5 min and the intracellular sul1 was reduced by 81.5 % after 30 min of the treatment. Compared to EC and PMS alone treatments, the conjugation transfer frequency of sul1 rapidly declined by 92.9 % within 2 min. The cell membrane, proteins, lipids, as well as intracellular and extracellular ARGs in E. coli DH5α were severely damaged by free radicals in solution and intracellular reactive oxygen species (ROS). Furthermore, up-regulation was observed in genes associated with oxidative stress, SOS response and cell membrane permeability in E. coli DH5α, however, no significant changes were observed in functional genes related to gene conjugation and transfer mechanisms. This study would contribute to the underlying of PMS activation by three-dimensional particle electrode, and provide novel insights into the mechanism of ARB inactivation and ARGs degradation under PMS advanced oxidation treatment.202439197284
8819170.9929Responses of Bacillus sp. under Cu(II) stress in relation to extracellular polymeric substances and functional gene expression level. The production and composition of extracellular polymeric substances (EPS), as well as the EPS-related functional resistance genes and metabolic levels of Bacillus sp. under Cu(II) stress, were investigated. EPS production increased by 2.73 ± 0.29 times compared to the control when the strain was treated with 30 mg L(-1) Cu(II). Specifically, the polysaccharide (PS) content in EPS increased by 2.26 ± 0.28 g CDW(-1) and the PN/PS (protein/polysaccharide) ratio value increased by 3.18 ± 0.33 times under 30 mg L(-1) Cu(II) compared to the control. The increased EPS secretion and higher PN/PS ratio in EPS strengthened the cells' ability to resist the toxic effect of Cu(II). Differential expression of functional genes under Cu(II) stress was revealed by Gene Ontology pathway enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. The enriched genes were most obviously upregulated in the UMP biosynthesis pathway, the pyrimidine metabolism pathway, and the TCS metabolism pathway. This indicates an enhancement of EPS regulation-related metabolic levels and their role as a defense mechanism for cells to adapt to Cu(II) stress. Additionally, seven copper resistance genes were upregulated while three were downregulated. The upregulated genes were related to the heavy metal resistance, while downregulated genes were related to cell differentiation, indicating that the strain had initiated an obvious resistance to Cu(II) despite its severe cell toxicity. These results provided a basis for promoting EPS-regulated associated functional genes and the application of gene-regulated bacteria in heavy metal-containing wastewater treatment.202337195605
7969180.9929Metagenomic insights into the influence of pH on antibiotic removal and antibiotic resistance during nitritation: Regulations on functional genus and genes. The changes in pH and the resulting presence of free nitrous acid (FNA) or free ammonia (FA) often inhibit antibiotic biodegradation during nitritation. However, the specific mechanisms through which pH, FNA and FA influence antibiotic removal and the fate of antibiotic resistance genes (ARGs) are not yet fully understood. In this study, the effects of pH, FNA, and FA on the removal of cefalexin and amoxicillin during nitritation were investigated. The results revealed that the decreased antibiotic removal under both acidic condition (pH 4.5) and alkaline condition (pH 9.5) was due to the inhibition of the expression of amoA in ammonia-oxidizing bacteria and functional genes (hydrolase-encoding genes, transferase-encoding genes, lyase-encoding genes, and oxidoreductase-encoding genes) in heterotrophs. Furthermore, acidity was the primary inhibitor of antibiotic removal at pH 4.5, followed by FNA. Antibiotic removal was primarily inhibited by alkalinity at pH 9.5, followed by FA. The proliferation of ARGs mediated by mobile genetic element was promoted under both acidic and alkaline conditions, attributed to the promotion of FNA and FA, respectively. Overall, this study highlights the inhibitory effects of acidity and alkalinity on antibiotic removal during nitritation.202439068965
8044190.9929Effect 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