Differential size-dependent response patterns and antibiotic resistance development mechanism in anammox consortia. - Related Documents




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795701.0000Differential size-dependent response patterns and antibiotic resistance development mechanism in anammox consortia. Antibiotic resistance is a global threat to human and animal health. Anaerobic ammonia oxidation (anammox) is an efficient and innovative wastewater treatment technology, which can be served as a promising approach to teat antibiotic wastewater. This study systematically investigated effects of sulfamethazine on the performance, microbial community dynamics and the resistome in anammox systems inoculated with different-sized granular sludge. The activity and performance of small (< 0.5 mm) anammox granules were more susceptible to sulfamethazine stress than those of medium (0.5-1.0 mm) and large (1.0-2.0 mm) granules. Sulfamethazine addition greatly increased the diversity and abundance of mobile genetic elements (MGEs) and antibiotic resistance genes (ARGs). Based on the metagenomic analysis, the horizontal transfer of ARGs in the anammox system was upregulated through bacterial oxidative stress, pili synthesis and type IV secretion system. In addition, two strains of sulfamethazine-resistant bacteria (Pseudomonas asiatica sp. nov. and Pseudomonas shirazica sp. nov.) were isolated from the anammox system. Their whole genome sequencing results showed that the most abundant plasmid was pkF7158B, which mediated the horizontal transfer of two main multidrug resistance genes (cpxR and mexB). This work provides a holistic insight into microbial heterogeneity of different-sized anammox granular sludge and their evolution and resistance development mechanism.202540086246
795810.9998Microbial response and recovery strategy of the anammox process under ciprofloxacin stress from pure strain and consortia perspectives. Ciprofloxacin (CIP) poses a high risk of resistance development in water environments. Therefore, comprehensive effects and recovery strategies of CIP in anaerobic ammonia oxidation (anammox) process were systematically elucidated from consortia and pure strains perspectives. The anammox consortia was not significantly affected by the stress of 10 mg L(-1) CIP, while the higher concentration (20 mg L(-1)) of CIP caused a dramatic reduction in the nitrogen removal performance of anammox system. Simultaneously, the abundances of dominant functional bacteria and corresponding genes also significantly decreased. Such inhibition could not be mitigated by the recovery strategy of adding hydrazine and hydroxylamine. Reducing nitrogen load rate from 5.1 to 1.4 kg N m(-3) d(-)(1) promoted the restoration of three reactors. In addition, the robustness and recovery of anammox systems was evaluated using starvation and shock strategies. Simultaneously, antibiotic resistance genes and key metabolic pathways of anammox consortia were upregulated, such as carbohydrate and energy metabolisms. In addition, 11 pure stains were isolated from the anammox system and identified through phylogenetic analysis, 40 % of which showed multidrug resistance, especially Pseudomonas. These findings provide deep insights into the responding mechanism of anammox consortia to CIP stress and promote the application of anammox process for treating wastewater containing antibiotics.202438554504
758520.9998Impacts of engineered nanoparticles and antibiotics on denitrification: Element cycling functional genes and antibiotic resistance genes. The wide presence of antibiotics and minerals warrants their combined effects on the denitrification in natural aquatic environment. Herein, we investigated the effects of two antibiotics, sulfamethazine (SMZ) and chlortetracycline (CTC), on the reduction of NO(3)(-)-N and accumulation of NO(2)(-)-N in the absence and presence of engineered nanoparticles (NPs) (Al(2)O(3), SiO(2), and geothite) using 16 S rRNA sequencing and high-throughput quantitative PCR. The results showed that the addition of antibiotics inhibited the reduction of NO(3)(-)-N by changing the bacterial community structure and reducing the abundance of denitrification genes, while engineered NPs promoted the denitrification by increasing the abundance of denitrification genes. In the binary systems, engineered NPs alleviated the inhibitory effect of antibiotics through enriching the denitrification genes and adsorbing antibiotics. Antibiotics and its combination with engineered NPs changed the composition of functional genes related to C, N, P, S metabolisms (p < 0.01). The addition of antibiotics and/or engineered NPs altered the bacterial community structure, which is dominated by the genera of Enterobacter (40.7-90.5%), Bacillus (4.9-58.5%), and Pseudomonas (0.21-12.7%). The significant relationship between denitrification, carbon metabolism genes, and antibiotic resistance genes revealed that the heterotrophic denitrifying bacteria may host the antibiotic resistance genes and denitrification genes simultaneously. The findings underscore the significance of engineered NPs in the toxicity assessment of pollutants, and provide a more realistic insight into the toxicity of antibiotics in the natural aquatic environment.202235738104
758430.9998Responses 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
851640.9998Graphene Oxide Inhibits Antibiotic Uptake and Antibiotic Resistance Gene Propagation. Antibiotics and antibiotic resistance genes (ARGs) in the natural environment have become substantial threats to the ecosystem and public health. Effective strategies to control antibiotics and ARG contaminations are emergent. A novel carbon nanomaterial, graphene oxide (GO), has attracted a substantial amount of attention in environmental fields. This study discovered the inhibition effects of GO on sulfamethoxazole (SMZ) uptake for bacteria and ARG transfer among microorganisms. GO promoted the penetration of SMZ from intracellular to extracellular environments by increasing the cell membrane permeability. In addition, the formation of a GO-SMZ complex reduced the uptake of SMZ in bacteria. Moreover, GO decreased the abundance of the sulI and intI genes by approximately 2-3 orders of magnitude, but the global bacterial activity was not obviously inhibited. A class I integron transfer experiment showed that the transfer frequency was up to 55-fold higher in the control than that of the GO-treated groups. Genetic methylation levels were not significant while sulI gene replication was inhibited. The biological properties of ARGs were altered due to the GO-ARG noncovalent combination, which was confirmed using multiple spectral analyses. This work suggests that GO can potentially be applied for controlling ARG contamination via inhibiting antibiotic uptake and ARG propagation.201627934199
759050.9997Characteristics, Performance and Microbial Response of Aerobic Granular Sludge for Treating Tetracycline Hypersaline Pharmaceutical Wastewater. Salt-tolerant aerobic granular sludge(AGS) was successfully cultivated under the dual stress of tetracycline and 2.5% salinity, resulting in an average particle size of 435.0 ± 0.5 and exhibiting a chemical oxygen demand(COD) removal rate exceeding 80%, as well as excellent sedimentation performance. The analysis of metagenomics technology revealed a significant pattern of succession in the development of AGS. The proportion of Oleiagrimonas, a type of salt-tolerant bacteria, exhibited a gradual increase and reached 38.07% after 42 days, which indicated that an AGS system based on moderate halophilic bacteria was successfully constructed. The expression levels of targeted genes were found to be reduced across the entire AGS process and formation, as evidenced by qPCR analysis. The presence of int1 (7.67 log10 gene copies g(-1) in 0 d sludge sample) enabled microbes to horizontally transfer ARGs genes along the AGS formation under the double pressure of TC and 2.5% salinity. These findings will enhance our understanding of ARG profiles and the development in AGS under tetracycline pressure, providing a foundation for guiding the use of AGS to treat hypersaline pharmaceutical wastewater.202438930555
763060.9997Coexistence of silver ion and tetracycline at environmentally relevant concentrations greatly enhanced antibiotic resistance gene development in activated sludge bioreactor. Antibiotic resistance has become a global public health problem. Recently, various environmental pollutants have been reported to induce the proliferation of antibiotic resistance. However, the impact of multiple pollutants (e.g., heavy metals and antibiotics), which more frequently occur in practical environments, is poorly understood. Herein, one widely distributed heavy metal (Ag(+)) and one frequently detected antibiotic (tetracycline) were chosen to investigate their coexisting effect on the proliferation of antibiotic resistance in the activated sludge system. Results show that the co-occurrence of Ag(+) and tetracycline at environmentally relevant concentrations exhibited no distinct inhibition in reactor performances. However, they inhibited the respiratory activity by 42%, destroyed the membrane structure by 218%, and increased membrane permeability by 29% compared with the blank control bioreactor. Moreover, the relative abundances of target antibiotic resistance genes (ARGs) (e.g., tetA, bla(TEM-1), and sulII) in effluent after exposure of coexisting Ag(+) and tetracycline were increased by 92-1983% compared with those in control reactor, which were 1.1-4.3 folds higher than the sum of the sole ones. These were possibly attributed to the enrichments of antibiotic-resistant bacteria. The results would illumine the coexisting effect of heavy metals and antibiotics on the dissemination of ARGs in activated sludge system.202234482077
796670.9997How heavy metal stress promotes dissemination of antibiotic resistance genes in the activated sludge process. Heavy metals have been recently revealed as promoters to antibiotic resistance gene (ARG) dissemination in water environment, but their influence on ARG transfer in the activated sludge process has not been clear. In this study, a set of sequencing batch reactors (SBRs) and micro-scale microfluidic chips were established to quantify the impacts of heavy metals (0.5 mM of Pb, 0.1 mM of As, and 0.005 mM of Hg) on the ARG spreading in the activated sludge consortium. Under heavy metal stress, transfer frequencies were 1.7-3.6 folds increase compared to the control. Gram-negative bacteria increased significantly after heavy metal added, which were more prone to receiving resistant plasmid from donors. Meanwhile, the relative expression of genes related to conjugation changed in activated sludge, especially the expression of outer membrane protein and oxidative stress regulatory genes increased by 2.9-7.4 folds and 7.8-13.1 folds, respectively. Furthermore, using microfluidic chips, the dynamics of ARG transfer was observed at single cell level under heavy metal pressure. Heavy metals firstly promoted conjugation and then vertical gene transfer played an important part for ARG spreading. The results provided in-depth understanding of the influence of heavy metals on ARG behavior in the environment.202235724617
795980.9997Evolution 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
758290.9997Anaerobic fermentation for hydrogen production and tetracycline degradation: Biodegradation mechanism and microbial community succession. The misuse and continues discharge of antibiotics can cause serious pollution, which is urgent to take steps to remit the environment pollution. In this study, anaerobic bacteria isolated from the aeration tank of a local sewage treatment plant were employed to investigate hydrogen production and tetracycline (TC) degradation during anaerobic fermentation. Results indicate that low concentrations of TC enhanced hydrogen production, increasing from 366 mL to a maximum of 480 mL. This increase is attributed to stimulated hydrolysis and acidogenesis, coupled with significant inhibition of homoacetogenesis. Furthermore, the removal of TC, facilitated by adsorption and biodegradation, exceeded 90 %. During the fermentation process, twenty-one by-products were identified, leading to the proposal of four potential degradation pathways. Analysis of the microbial community revealed shifts in diversity and a decrease in the abundance of hydrogen-producing bacteria, whereas bacteria harboring tetracycline resistance genes became more prevalent. This study provides a possibility to treat tetracycline-contaminated wastewater and to produce clean energy simultaneously by anaerobic fermentation.202439168318
7583100.9997Insights into the combined effect of ofloxacin and humic substances on sewage sludge anaerobic digestion. Humic substances (HS) and antibiotics are present simultaneously in various environments. However, the influence path and consequences of HS on antibiotics behaviors in complex anaerobic microbial systems are rarely known, hindering the understanding and control of antibiotics risks. This study for the first time investigated the combined effects of ofloxacin (OFL) and HS in sewage sludge anaerobic digestion system. Experimental results showed that OFL alone reduced the cumulative methane production and the maximum methane production rate by 14.6 % and 33.5 %, respectively. The methane production curves showed step by step adaption, which might be related with the increase of antibiotics resistance genes and their potential hosts. The coexistence of low concentration (6 % of sludge volatile solid) HS could alleviate the inhibition of OFL on hydrolysis-related bacteria and genes to a certain extent, thereby enhanced the methane production by 4.8 %. However, the coexistence of high concentration (12-24 % of sludge volatile solid) HS intensified the inhibition on hydrolysis-related bacteria and genes, and had more potential to combine with organic matters to prevent sludge solubilization, macromolecular organics hydrolysis and OFL degradation, thereby further decreasing the methane production by 7.6-15.9 %. Besides, the coexistence of OFL and high concentration HS increased the antimicrobial resistance and pathogenicity risks of digested sludge, by enhancing the residual level of verified pathogens, antibiotics resistance genes and virulence factor genes. This study provides new insights into the environmental risks of combined antibiotics and HS pollution, and offers a basis for strengthening the safe treatment and disposal of sewage sludge.202540752562
7560110.9997The 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
7581120.9997Enhanced performance of anaerobic digestion of cephalosporin C fermentation residues by gamma irradiation-induced pretreatment. Antibiotic fermentation residues is a hazardous waste due to the existence of residual antibiotics and antibiotic resistance genes (ARGs), probably leading to the induction and spread of antibiotic resistant bacteria (ARB) in the environment, which could pose potential harm to the ecosystem and human health. It is urgent to develop an effective technology to remove the residual antibiotics and ARGs. In this study, the anaerobic digestion combined with gamma irradiation was applied for the disposal and utilization of cephalosporin C fermentation residues. The experimental results showed that the antibacterial activities of cephalosporin C against Staphylococcus aureus were significantly decreased after anaerobic digestion. The removal of tolC, a multidrug resistant gene, was improved up to 100% by the combination of gamma irradiation and anaerobic digestion compared to solely anaerobic digestion process, which may be due to the changes of microbial community structures induced by gamma irradiation.202031590081
7589130.9997Neglected risks of enhanced antimicrobial resistance and pathogenicity in anaerobic digestion during transition from thermophilic to mesophilic. Minimization of antibiotic resistance genes (ARGs) and potential pathogenic antibiotic-resistant bacteria (PARB) during anaerobic digestion (AD) is significantly impacted by temperature. However, knowledge on how ARGs and PARB respond to temperature transition from thermophilic to mesophilic is limited. Here, we combined metagenomic-based with culture-based approaches and revealed the risks of antimicrobial resistance and pathogenicity during transition from 55 °C to 35 °C for AD, with strategies of sharp (ST, one-step by 20 °C/d) and mild (MT, step-wise by 1 °C/d). Results indicated a lower decrease in methane production with MT (by 38.9%) than ST (by 88.8%). Phenotypic assays characterized a significant propagation of multi-resistant lactose-fermenting Enterobacteriaceae and indicator pathogens after both transitions, especially via ST. Further genomic evidence indicated a significant increase of ARGs (29.4-fold), virulence factor genes (1.8-fold) and PARB (65.3-fold) after ST, while slight enrichment via MT. Bacterial succession and enhanced horizontal transfer mediated by mobile genetic elements promoted ARG propagation in AD during transition, which was synchronously exacerbated through horizontal transfer mechanisms mediated by cellular physiological responses (oxidative stress, membrane permeability, bacterial conjugation and transformation) and co-selection mechanisms of biomethanation metabolic functions (acidogenesis and acetogenesis). This study reveals temperature-dependent resistome and pathogenicity development in AD, facilitating microbial risk control.202438878435
7629140.9997Graphene oxide in the water environment could affect tetracycline-antibiotic resistance. In recent years, the influence of new materials like nanoparticles in the water environment on biological substances has been widely studied. Antibiotic resistance genes (ARGs) represent a new type of pollutant in the environment. Graphene oxide (GO), as a nano material, because of its unique structure, may have an impact on antibiotic resistance bacteria (ARB) and ARGs; however the research in this area is rarely reported. Therefore, this study mainly investigated the effects of GO on bacterial antibiotic resistance. The results showed that GO had a limited effect on ARB inactivation. A high concentration of GO (>10 mg/L) can damage resistant plasmids to reduce bacterial resistance to antibiotics, but low concentrations of GO (<1 mg/L) led to almost no damage to the plasmid. However, all tested concentrations of GO promoted the conjugative transfer from 1to over 3 folds, with low concentrations and high concentration (1-10 and 100 mg/L) of GO samples the least promoted. The overall effect of GO on antibiotic resistance needs further investigation.201728549325
7598150.9997Aspartame affects methane yield and enhances transmission of antibiotic resistance genes during anaerobic digestion of sludge. Aspartame (ASP) is a widely used artificial sweetener, yet recent studies have shown that ASP have potential toxic effect. ASP is also detected in sludge, however, the influence of ASP on the performance of sludge anaerobic digestion and the fate of antibiotic resistance genes (ARGs) have not been thoroughly investigated. Under stress of 0, 0.5, 5 and 50 mg/L ASP, cumulative methane production was 181.7, 167.0, 154.0 and 140.8 mlCH(4)/g VSS, respectively. ASP inhibited the dissolution and conversion of organic matter in sludge. Sequencing data revealed a decline in the abundance of functional microorganisms compared to control, such as hydrolytic-acidifying bacteria and methanogens, potentially attributed to increased intracellular reactive oxygen species and damaged cell membranes caused by ASP addition. Specifically, 50 mg/L ASP reduced the total abundance of methanogens by 59.40 % compared to control. Concurrently, alterations in microbial communities along with an increase in Tn916 and intI1 were observed, increasing the abundance of ARGs. The total abundance of five ARGs peaked at 1.43E+ 12 copies/g at 5 mg/L ASP, representing 139 % of the control. This research contributes valuable insights into the alterations in anaerobic digestion caused by ASP, emphasizing the potential risks in the overall environmental system.202539616849
7960160.9997Diversity evolution of functional bacteria and resistance genes (CzcA) in aerobic activated sludge under Cd(II) stress. An activated sludge sequencing batch reactor (SBR) was used to treat divalent cadmium (Cd(II)) wastewater for 60 d to investigate the overall treatment performance, evolution of the bacterial community, and abundance of the Cd(II) resistance gene CzcA and shifts in its potential host bacteria. During stable operation with a Cd(II) concentration of 20 mg/L, the average removal efficiencies of Cd(II) and chemical oxygen demand (COD) were more than 85% and that of total phosphorus was greater than 70%, while the total nitrogen (TN) was only about 45%. The protein (PN) content in the extracellular polymeric substances (EPS) increased significantly after Cd(II) addition, while polysaccharides displayed a decreasing trend (p < 0.05), indicating that EPS prefer to release PN to adsorb Cd(II) and protect bacteria from damage. Three-dimensional fluorescence spectral analysis showed that fulvic acid-like substances were the most abundant chemical components of EPS. The addition of Cd(II) adversely affected most denitrifying bacteria (p < 0.05), which is consistent with the low TN removal. In addition, quantitative polymerase chain reaction analysis revealed that CzcA gene abundance decreased as the Cd(II) concentration increased, possibly because expression of the CzcA gene was inhibited by Cd(II) stress. The majority of CzcA gene sequences were carried by Pseudomonas, making it the dominant genus among Cd(II)-resistant bacteria.201931514000
7561170.9997Deciphering antibiotic resistome variations during nitrogen removal process transition under mixed antibiotics stress: Assembly process and driving factors. Antibiotic resistome, which encompasses all types of antibiotic resistance genes (ARGs) in a given environment, has received increasing attention in research on different wastewater treatment processes. However, the variation in antibiotic resistome during the transition from the full nitrification-denitrification to the shortcut nitrification-denitrification process remains unclear. In this study, a total of 269 targeted gene subtypes were identified, along with 108 genes were consistently present in all samples. The introduction of mixed antibioticsrapidly increased the abundance of corresponding and non-corresponding ARGs, as well as that of mobile genetic elements.The variations in of the antibiotic resistome were primarily driven by dissolved oxygen and nitrite accumulation rate. Moreover, 34 bacterial genera were identified as potential ARG hosts, with most denitrifiers considered as potential antibiotic-resistant bacteria, including Branchymonas, Rhodobacter, and Thauera. This study provides a method for controlling antibiotic resistance by regulating the changes in environmental variables and bacterial communities.202339492537
8517180.9997Influences of graphene on microbial community and antibiotic resistance genes in mouse gut as determined by high-throughput sequencing. Graphene is a promising candidate as an antibacterial material owning to its bacterial toxicity. However, little information on influence of graphene on gut microbiota is available. In this study, mice were exposed to graphene for 4 weeks, and high-throughput sequencing was applied to characterize the changes in microbial community and antibiotic resistance genes (ARGs) in mouse gut. The results showed that graphene exposure increased biodiversity of gut microbiota, and changed their community. The 1 μg/d graphene exposure had higher influences on the gut microbiota than 10 μg/d and 100 μg/d graphene exposures, which might be due to higher aggregation of high-level graphene. The influence of graphene on gut microbiota might attribute to that graphene could induce oxidative stress and damage of cell membrane integrity. The results were verified by the increase of ratio of Gram-negative bacteria. Outer membrane of Gram-negative bacteria could reduce the membrane damage induced by graphene and make them more tolerance to graphene. Further, we found that graphene exposure significantly increased the abundance and types of ARGs, indicating a potential health risk of graphene. This study firstly provides new insight to the health effects of graphene on gut microbiota.201626476051
8503190.9997Dual-pathway inhibition of antibiotic resistance genes by ferrate (Fe(VI)): Oxidative inactivation and genetic mobility impairment in anaerobically digested sludge. Antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB) are emerging environmental contaminants that threaten public health, highlighting the urgent need for effective control strategies. Ferrate (Fe(VI)), a strong and eco-friendly oxidant, shows great potential for this purpose. This study systematically evaluated the efficacy of Fe(VI) in mitigating ARGs and ARB in anaerobically digested sludge, with a particular focus on elucidating the underlying mechanisms by which Fe(VI) effects ARGs dissemination through both vertical gene transfer (VGT) and horizontal gene transfer (HGT). Result shows that Fe(VI) doses of 20 and 60 mg/g-TS reduce ARGs by 9.75 % and 19.12 %, respectively, while inactivating up to 24.7 % of ARB at the higher dose. Pathogenic ARB, such as Escherichia coli and Shigella sonnei, are preferentially removed, with abundances decrease by 63.7 % and 28.0 %. Mechanistically, the structural disruption of bacterial cells caused by Fe(VI) in anaerobically digested sludge, as indicated by a 29 % reduction in extracellular polymeric substances and a 23.7 % increase in cell membrane permeability. Subsequently, a marked release of intracellular ARGs into the extracellular environment is also observed, where they are likely subjected to degradation by Fe(VI). This oxidative killing accounts for the observed ARB decrease, thereby limiting the VGT of ARGs. In addition, Fe(VI) impairs the HGT of ARGs by diminishing their mobility potential, reflected in the reduced co-occurence with mobile genetic elements. Meanwhile, sludge bacterial competence for DNA uptake and recombination is markedly reduced, as evidenced by a 9.8 % decline in the abundance of related functional genes. These findings demonstrate that Fe(VI) effectively inhibits the dissemination of ARGs by targeting both primary transmission pathways. It suppresses VGT, thereby reducing the inheritance of ARB within populations, and limits HGT, curbing the spread of mobile ARGs among competent species. By disrupting these two critical routes, Fe(VI) shows strong potential as an effective strategy for mitigating ARGs propagation in sludge systems.202541138327