Fate of antibiotic resistance genes and resistant bacteria under various operating temperatures of sludge anaerobic digestion. - Related Documents




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755901.0000Fate of antibiotic resistance genes and resistant bacteria under various operating temperatures of sludge anaerobic digestion. This study investigates the impact of varying temperatures on reducing antibiotic resistance genes (ARGs) during anaerobic digestion (AD) of mixed raw sludge in wastewater treatment plants. Employing three different operating temperatures, i.e., 37, 55, and 65 °C, the research aims to identify how these conditions affect the diminution of resistant genes. The results, based on quantitative PCR analysis and metagenomic sequencing, show that higher temperatures significantly enhance the reduction of ARGs, with the most substantial decreases observed at 65 °C. This temperature-dependent reduction correlates with changes in the microbial community structure, where specific bacterial genera like Alicycliphilus, Macellibacteroides, Dokdonella, Ahniella, Thauera, and Zoogloea associated with ARGs exhibit decreased abundance at elevated temperatures. The study infers that AD at higher temperatures could be a more effective strategy in mitigating the spread of antibiotic resistance in the environment, suggesting a pivotal role of operational temperature in optimizing wastewater treatment processes for ARGs attenuation. The findings highlight the need for further research to refine AD protocols, aiming to minimize the environmental impact of antibiotic resistance dissemination.202540662898
755110.9999Microbial community evolution and fate of antibiotic resistance genes during sludge treatment in two full-scale anaerobic digestion plants with thermal hydrolysis pretreatment. Anaerobic digestion (AD) with thermal hydrolysis pretreatment is widely used as an efficient sludge treatment nowadays. However, the evolution of microbial community (especially for the archaea community), the fate of antibiotic resistance genes (ARGs), and their associations during such process in full-scale sludge treatment plants are rarely reported. Therefore, these scientific questions were explored at two full-scale sludge treatment plants through high-throughput sequencing and quantitative PCR. Results showed that Methanobacterium and Methanosphaera were the dominant archaea in thermal hydrolyzed sludge. The predominant bacteria in the sludge first shifted from nutrients removal functional bacteria to spore-forming bacteria after thermal hydrolysis, and then shifted to fermentative bacteria after AD. The full-scale plants could select ermB, ermF, mefA/E, qnrS and tetM. Though the bacteria and archaea biomass and community largely influenced the fate of ARGs, multiple linear regression analysis showed that the total ARGs were mainly affected by mobile genetic elements (MGEs).201931158777
703420.9999Meta-analysis reveals the processes and conditions of using biochar to control antibiotic resistance genes in soil. Soil is a significant reservoir of antibiotic resistance genes (ARGs) and an important habitat for pathogens associated with many clinical infections and plant disease outbreaks. Although scientists have found that biochar can reduce ARGs in soil, the understanding of how biochar removes soil ARGs and the influencing factors remains limited. Here, a meta-analysis of 65 published studies was conducted to illuminate the mechanisms through which biochar remediates ARG-contaminated soils. In biochar-amended soil, the antibiotic content significantly decreased by 24.1 %, while the abundances of mobile genetic elements and ARG host bacteria declined by 23.5 % and 12.1 %, respectively. The reduced antibiotic content, suppressed mobile genetic elements, and altered bacterial community structure collectively led to a 41.8 % reduction in soil ARG abundance. In addition, wood-derived biochar pyrolyzed at 300-500 °C exhibited a substantial advantage in the remediation of ARGs. Furthermore, biochar application decreased the abundance of ARGs in alkaline and neutral soil more markedly than that in acidic soil. The results of this research confirmed the positive mitigating effect of biochar on ARGs in soil, providing valuable insights for the prevention and control of ARG pollution.202540359860
702330.9999Metagenomic absolute quantification of antibiotic resistance genes and virulence factor genes-carrying bacterial genomes in anaerobic digesters. Sewage treatment works have been considered as hotspots for the dissemination of antibiotic resistance genes (ARGs). Anaerobic digestion (AD) has emerged as a promising approach for controlling the spread of ARGs while destroying biomass in sludge. Evaluating the impact of AD on ARG removal relies on the absolute quantification of ARGs. In this study, we quantified the ARG concentrations in both full-scale and lab-scale AD systems using a cellular spike-ins based absolute quantification approach. Results demonstrated that AD effectively removed 68 ± 18 %, 55 ± 12 %, and 57 ± 19 % of total ARGs in semi-continuous AD digesters, with solid retention times of 15, 20, and 25 days, respectively. The removal efficiency of total ARGs increased as the AD process progressed in the batch digesters over 40 days. A significant negative correlation was observed between digestion time and the concentrations of certain ARG types, such as beta-lactam, sulfonamide, and tetracycline. However, certain potential pathogenic antibiotic resistant bacteria (PARB) and multi-resistant high-risk ARGs-carrying populations robustly persisted throughout the AD process, regardless of the operating conditions. This study highlighted the influence of the AD process and its operating parameters on ARG removal, and revealed the broad spectrum and persistence of PARB in AD systems. These findings provided critical insights for the management of microbial hazards.202438359594
703040.9999Metagenomic profiling of antibiotic resistance genes/bacteria removal in urban water: Algal-bacterial consortium treatment system. Antibiotic resistance genes (ARGs) have exhibited significant ecological concerns, especially in the urban water that are closely associated with human health. In this study, with presence of exogenous Chlorella vulgaris-Bacillus licheniformis consortium, most of the typical ARGs and MGEs were removed. Furthermore, the relative abundance of potential ARGs hosts has generally decreased by 1-4 orders of magnitude, revealing the role of algal-bacterial consortium in cutting the spread of ARGs in urban water. While some of ARGs such as macB increased, which may be due to the negative impact of algicidal bacteria and algal viruses in urban water on exogenous C. vulgaris and the suppression of exogenous B. licheniformis by indigenous microorganisms. A new algal-bacterial interaction might form between C. vulgaris and indigenous microorganisms. The interplay between C. vulgaris and bacteria has a significant impact on the fate of ARGs removal in urban water.202438801952
755250.9999Effect of temperature on removal of antibiotic resistance genes by anaerobic digestion of activated sludge revealed by metagenomic approach. As antibiotic resistance continues to spread globally, there is growing interest in the potential to limit the spread of antibiotic resistance genes (ARGs) from wastewater sources. In particular, operational conditions during sludge digestion may serve to discourage selection of resistant bacteria, reduce horizontal transfer of ARGs, and aid in hydrolysis of DNA. This study applied metagenomic analysis to examine the removal efficiency of ARGs through thermophilic and mesophilic anaerobic digestion using bench-scale reactors. Although the relative abundance of various ARGs shifted from influent to effluent sludge, there was no measureable change in the abundance of total ARGs or their diversity in either the thermophilic or mesophilic treatment. Among the 35 major ARG subtypes detected in feed sludge, substantial reductions (removal efficiency >90%) of 8 and 13 ARGs were achieved by thermophilic and mesophilic digestion, respectively. However, resistance genes of aadA, macB, and sul1 were enriched during the thermophilic anaerobic digestion, while resistance genes of erythromycin esterase type I, sul1, and tetM were enriched during the mesophilic anaerobic digestion. Efflux pump remained to be the major antibiotic resistance mechanism in sludge samples, but the portion of ARGs encoding resistance via target modification increased in the anaerobically digested sludge relative to the feed. Metagenomic analysis provided insight into the potential for anaerobic digestion to mitigate a broad array of ARGs.201525994259
808660.9999Biofilm 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
702570.9998Aerobic composting as an effective cow manure management strategy for reducing the dissemination of antibiotic resistance genes: An integrated meta-omics study. Livestock manure is considered as an important source for spreading antibiotic resistance genes (ARGs) into the environment, and therefore poses a direct threat to public health. Whereas the effects of reused manure on soil microbial communities and ARGs have been studied extensively, comprehensive characterizations of microbial communities and ARGs of manure produced by different management methods are not well understood. Here, we analyzed the fate of microbial communities and ARGs of cow manure treated by three conventional management strategies: aerobic composting, mechanical drying and precipitation, applying an integrated-omics approach combining metagenomics and metaproteomics. Integrated-omics demonstrated that composted manure contained the lowest diversity of microbial community and ARGs compared with manure treated by other two strategies. Quantitative PCR methods revealed that the abundances of ARGs were reduced by over 83 % after composting for 14 days, regardless of the season. Besides, the potential ARG hosts Acinetobacter and Pseudomonas dominating mechanical drying process were sharply decreased in abundances after composting. The significant co-occurrence networks among bacteria, ARGs and transposase gene tnpA-01 in composting samples indicated the important role of these bacteria in the dissemination of ARGs. These findings offer insight into potential strategies to control the spread of ARGs during livestock manure reuse.202031884359
857880.9998Impact of earthworms on suppressing dissemination of antibiotic resistance genes during vermicomposting treatment of excess sludge. Earthworms play a crucial role in suppressing the dissemination of antibiotic resistance genes (ARGs) during vermicomposting. However, there is still a lack of how earthworms influence the spread of ARGs. To address this gap, a microcosm experiment was conducted, incorporating earthworms and utilizing metagenomics and quantitative PCR to assess the impact of earthworms on microbial interactions and the removal of plasmid-induced ARGs. The findings revealed that vermicomposting led to a reduction in the relative abundance of ARGs by altering microbial communities and interactions. Significantly, vermicomposting demonstrated an impressive capability, reducing 92% of ARGs donor bacteria and impeding the transmission of 94% of the RP4 plasmid. Furthermore, through structural equation model analysis, it was determined that mobile genetic elements and environmental variables were the primary influencers of ARG reduction. Overall, this study offers a fresh perspective on the effects of vermicomposting and its potential to mitigate the spread of ARGs.202438885722
702990.9998Filamentous bacteria-induced sludge bulking can alter antibiotic resistance gene profiles and increase potential risks in wastewater treatment systems. Sludge bulking caused by filamentous bacteria is a prevalent issue in wastewater treatment systems. While previous studies have primarily concentrated on controlling sludge bulking, the biological risks associated with it have been overlooked. This study demonstrates that excessive growth of filamentous bacteria during sludge bulking can significantly increase the abundance of antibiotic resistance genes (ARGs) in activated sludge. Through metagenomic analysis, we identified specific ARGs carried by filamentous bacteria, such as Sphaerotilus and Thiothrix, which are responsible for bulking. Additionally, by examining over 1,000 filamentous bacterial genomes, we discovered a diverse array of ARGs across different filamentous bacteria derived from wastewater treatment systems. Our findings indicate that 74.84% of the filamentous bacteria harbor at least one ARG, with the occurrence frequency of ARGs in these bacteria being approximately 1.5 times higher than that in the overall bacterial population in activated sludge. Furthermore, genomic and metagenomic analyses have shown that the ARGs in filamentous bacteria are closely linked to mobile genetic elements and are frequently found in potentially pathogenic bacteria, highlighting potential risks posed by these filamentous bacteria. These insights enhance our understanding of ARGs in activated sludge and underscore the importance of risk management in wastewater treatment systems.202439094405
7560100.9998The 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
7032110.9998Free Radicals on Aging Microplastics Regulated the Prevalence of Antibiotic Resistance Genes in the Aquatic Environment: New Insight into the Effect of Microplastics on the Spreading of Biofilm Resistomes. The spread of antibiotic resistance genes (ARGs) by microplastics has received a great concern in coexisting "hotspots". Despite most microplastics suffering from natural aging, little is known about the effect of aging microplastics (A-MPs) on ARGs dissemination. Here, we demonstrated significant suppression of A-MPs on ARGs dissemination in natural rivers. Although ARGs and mobile genetic elements (MGEs) were effectively enriched on A-MPs, the relative abundance of ARGs and MGEs on A-MPs as well as in receiving water decreased by approximately 21.4% to 42.3% during a period of 30 days of dissemination. Further investigation revealed that (•)OH was consistently generated on A-MPs with a maximum value of 0.2 μmol/g. Importantly, scavenging of (•)OH significantly increased the relative abundance of ARGs and MGEs both on A-MPs and in receiving water 1.4-29.1 times, indicating the vital role of (•)OH in suppressing ARGs dissemination. Microbial analysis revealed that (•)OH inhibited the potential antibiotic-resistant bacteria in surface biofilms, such as Pseudomonas and Acinetobacter (with a decrease of 68.8% and 89.3%). These results demonstrated that (•)OH was extensively produced on A-MPs, which greatly reduced both the vertical and horizontal gene transfer of ARGs. This study provided new insights into the dissemination of ARGs through microplastics in natural systems.202540359213
7040120.9998The maturity period is the main stage of antibiotic resistance genes reduction in aerobic composting process of swine manure in sub-scale farms. This study was focused on the changes of antibiotic resistance genes (ARGs) and their potential host bacteria during the swine manure composting on sub-scale farms. Eight target ARGs increased 427% on average, with a trend of increase at early stage and decrease at later stage, and the main reduction stage appeared in maturity stage. The abundance of ARGs was mainly affected by the community succession of potential host bacteria. Composting could reduce the abundance of potential host bacteria of ARGs as well as pathogens such as Pseudomonas, and reduce the environmental risks of swine manure. N/C and S levels had a positive effect on the potential host of most ARGs. Prolonging the maturity period would inhibit the growth of potential host bacteria of ARGs during composting, therefore inhibiting the transmission of ARGs.202132971337
7561130.9998Deciphering 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
6950140.9998Ceftiofur in swine manure contributes to reducing pathogens and antibiotic resistance genes during composting. Aerobic composting is a common way for the disposal of feces produced in animal husbandry, and can reduce the release of antibiotic resistance genes (ARGs) from feces into the environment. In this study, we collected samples from two distinct treatments of swine manure compost with and without ceftiofur (CEF), and identified the ARGs, mobile genetic elements (MGEs), and bacterial community by metagenomic sequencing. The impacts of CEF on the bacterial community composition and fate of ARGs and MGEs were investigated. With increasing composting temperature and pH, the concentration of CEF in the manure decreased rapidly, with a degradation half-life of 1.12 d and a 100% removal rate after 10 d of aerobic composting. Metagenomics demonstrated that CEF in the manure might inhibit the growth of Firmicutes and Proteobacteria, thereby reducing some ARGs and MGEs hosted by these two bacteria, which was further confirmed by the variations of ARGs and MGEs. A further redundancy analysis suggested that pH and temperature are key environmental factors affecting ARG removal during composting, and intI1 and bacterial communities also have significant influence on ARG abundance. These results are of great significance for promoting the removal of some ARGs from animal manure by controlling some key environmental factors and the type of antibiotics used in animals.202438685300
7021150.9998Metagenomic insights into dissemination of antibiotic resistance across bacterial genera in wastewater treatment. The aim of this study was to evaluate the impacts of conventional wastewater treatment processes including secondary treatment and chlorination on the removal of antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB), and to assess the association of ARGs with their potential hosts in each treatment process. The results showed chlorination with subinhibitory concentration (<8 mg/L) resulted in an increased ARB number in the disinfection effluent. qPCR analysis indicated secondary treatment increased relative abundance of ARGs in remaining bacteria whereas disinfection reduced the relative abundance of those genes effectively. Metagenomic analysis revealed a significant shift of dominating bacterial genera harboring ARGs. Along the treatment train, 48, 95 and 80 genera were identified to be the ARG carriers in primary effluent, secondary effluent, and disinfection effluent, respectively. It was also found that secondary treatment increased the diversity of potential ARG hosts while both secondary treatment and chlorination broadened the host range of some ARGs at the genus level, which may be attributed to the spread of antibiotic resistance across bacterial genera through horizontal transfer. This study highlights the growing concerns that wastewater treatment plants (WWTPs) may disseminate ARGs by associating this effect to specific treatment stages and by correlating ARGs with their bacterial hosts.202133453487
7549160.9998Fate of antibiotic resistance genes during sludge anaerobic fermentation: Roles of different sludge pretreatment. Excess sludge, the primary by-product of wastewater treatment plants, is the source and sink of antibiotic resistance genes (ARGs). Sludge pretreatments are an indispensable pathway to improve the resource recovery and harmfulness for anaerobic digestion sludge. However, fewer studies have compared the effects of different pretreatment technologies on the distribution of ARGs during anaerobic sludge digestion. Here, this study established seven anaerobic digesters, and four typical ARGs and one integrase gene of class 1 integron (intI1) regarded as the representative mobile genetic elements (MGEs) were examined during the whole anaerobic digestion process. It was found anaerobic digestion could effectively remove ARGs with about 70.86% removal rate of total ARGs. Among these pretreatments, the reduce efficiency of ARGs was the highest in 50 °C pretreatment, followed by oxidant, and the last was acid-alkaline. The microbial community analysis demonstrated the microbial community structure, including ARGs hosts and antibiotic resistant bacteria, was significantly changed and influenced by high temperature pretreatment. In addition, high temperature and K(2)S(2)O(8) observably decrease the level of ROS production. Macro transcriptome analysis indicated that sludge pretreatment, except for 50 °C pretreatment, up-regulated the genes relevant to lyases and transferase, but down-regulated the genes responsible for peroxidase, antioxidant enzymes and T4SS gene. This study emphasized and compared the different sludge pretreatments on the fate of ARGs in anaerobic sludge, and highlighted concerns regarding the environmental and health risks to our society.202439393457
6954170.9998Temporal effects of repeated application of biogas slurry on soil antibiotic resistance genes and their potential bacterial hosts. Biogas slurry, a liquid end product of animal manure fermentation, is widely used as fertilizer in crop fields. Land application may introduce antibiotics and related resistance genes from livestock production into agricultural soil. Nevertheless, changes in antimicrobial resistance in soil where biogas slurry has been repeatedly applied are not fully understood. In the present study, 13 veterinary antibiotics were analyzed in soils that were repeatedly sprayed with biogas slurry, and simultaneously, temporal changes in antibiotic resistance genes (ARGs) and bacterial community composition were investigated using a real-time quantitative PCR assay and MiSeq sequencing. Long-term repeated application of biogas slurry did not result in excessive accumulation of antibiotic residuals in the soil but increased the abundance of ARGs and facilitated ARG transfer among potential hosts. Although the quantitative PCR assay showed a decreasing trend for the relative abundance of ARGs over time, a relevance network analysis revealed highly complex bacteria-ARG co-occurrence after long-term application, which implied that repeated application might intensify horizontal gene transfer (HGT) of ARGs among different bacterial hosts in soil. The increased relative abundance of the intl1 gene supported the shift in ARG-bacteria co-occurrence. Furthermore, ordination analysis showed that the distributions of antibiotic resistance bacteria (ARB) and ARGs were closely related to application duration than to the influence of antibiotic residuals in the biogas slurry-treated soil environment. Additionally, natural level of ARG abundance in untreated soils indirectly suggested the presence/absence of antibiotics was not a key determinant causing the spread of antimicrobial resistance. This study provides improved insight into the effects of long-term repeated application of biogas slurry on the shift in ARG abundances and bacteria-ARG co-occurrence in soils, highlighting the need to focus on the influence of changed soil environment on the ARG transfer.202031818620
8083180.9998Alteration of the migration trajectory of antibiotic resistance genes by microplastics in a leachate activated sludge system. The environmental behavior of emerging contaminants of microplastics (MPs), antibiotics and antibiotic resistance genes (ARGs) in the leachate activated sludge system has been monitored and analyzed comprehensively. The results suggested that MPs could effectively alter the migration trajectory of tetracycline resistance genes (tet genes) in the leachate activated sludge system under intermittent and continuous influent conditions. After adding MPs, the total average abundance of tet genes in leachate increased from 0.74 ± 0.07 to 0.78 ± 0.07 (log(10)tet genes/log(10) 16S rRNA) and that in sludge increased from 0.65 ± 0.08 to 0.70 ± 0.06 (log(10)tet genes/log(10) 16S rRNA). Except for tetA, the abundance of tetB, tetO, tetM and tetQ on MPs increased with increasing TC concentration under both aerobic and anaerobic conditions. MPs not only significantly affect the abundance level and migration trajectory of ARGs in the leachate activated sludge system, but also remarkably improve the level of heavy metals in the ambient environment, indirectly promoting the selective effect of antibiotic-resistant bacteria (ARB) and promoting the development of antibiotic resistance (AR). In addition, MPs changed their physicochemical properties and released hazardous substances with aging to force tet genes to migrate from the leachate activated sludge system to the MPs, making AR more difficult to eliminate and persisted in wastewater treatment plants. Meanwhile, microorganisms played a driving role, making MPs serve as a niche for ARGs and ARB colonization. The co-occurrence network analysis indicated the specific distribution pattern of tet genes and microorganisms in different media, and the potential host was speculated. This study improves the understanding of the environmental behavior of emerging contaminants in leachate activated sludge system and lays a theoretical for protecting the ecological environment.202337321316
7022190.9998Fate of antibiotics, antibiotic-resistant bacteria, and cell-free antibiotic-resistant genes in full-scale membrane bioreactor wastewater treatment plants. The removal of antibiotics, antibiotic-resistant bacteria (ARB), and cell-free antibiotic-resistant genes (ARGs) and the microbial community of ARB were investigated in detail to understand their fate and provide valuable information on the feasibility of full-scale membrane bioreactor (MBR). The potential risks of cell-free ARGs to the receiving environment were discovered. High influent antibiotic concentration could inhibit the microbial activity of MBR sludge, whereas good antibiotic removal could be maintained because of relatively long solid retention time and high biomass retention. Approximately 61.8%-77.5% of the total antibiotics were degraded, and 22.5%-38.2% of the total antibiotics were adsorbed by MBR sludge on average. The individual antibiotic removal presented intense discrepancy because of the chemical construction and distribution coefficient of antibiotics. Aeromonas exhibited specific antibiotic resistance to ampicillin and erythromycin, Escherichia became the predominant genera in kanamycin-ARB and tetracycline-ARB, and Klebsiella and Bacteroides were the particular genera that exhibited distinct antibiotic resistance to ciprofloxacin. A significant correlation was found between cell-free ARG abundance and ARB content, and relatively high effluent cell-free ARG abundance facilitated the proliferation and transmission of ARB. The impacts of the receiving environment to eliminate the ecological risks and severe threats to human health should be investigated because of the low decay ratio and long-term persistence of cell-free ARGs.202031986335