Sulfadiazine degradation in soils: Dynamics, functional gene, antibiotic resistance genes and microbial community. - Related Documents




#
Rank
Similarity
Title + Abs.
Year
PMID
012345
755701.0000Sulfadiazine degradation in soils: Dynamics, functional gene, antibiotic resistance genes and microbial community. Sulfonamides and their corresponding antibiotic resistance genes (ARGs) are widespread in the environment, which leads to a major threat to global health crisis. Biodegradation plays a major role in sulfonamides removal in soil ecosystem, but the degradation dynamics and the associated functional bacteria in situ remain unclear. In this study, aerobic degradation of sulfadiazine (SDZ) at two dosages (1 and 10 mg/kg) was explored for up to 70 days in two different agricultural soils. The removal of SDZ in all treatments followed first-order multi-compartment model with half-life times of 0.96-2.57 days, and DT50 prolonged with the increase of initial dosage. A total of seven bacterial genera, namely Gaiella, Clostrium_sensu_stricto_1, Tumebacillus, Roseiflexus, Variocorax, Nocardioide and Bacillus, were proposed as the potential SDZ-degraders. sadA gene was for the first time detected in soil samples, but other functional genes might also participate in SDZ degradation. The enrichment of sulfonamide resistance genes was found after 70 days' incubation, which might result in the spread of ARGs in soil. This study can add some new insights towards SDZ degradation in soil ecosystem and provide a potential resource for the bioremediation of SDZ-contaminated soil.201931466189
808610.9997Biofilm 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
808420.9997Metagenomic 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
703030.9997Metagenomic 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
808240.9997Deciphering 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
807150.9997Impact 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
756060.9997The 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
756370.9997Polystyrene 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
755880.9997Interaction of sulfate-reducing bacteria and methanogenic archaea in urban sewers, leads to increased risk of proliferation of antibiotic resistance genes. Sewers are considered a potential reservoir of antibiotic resistance. However, the generation of antibiotic resistance genes (ARGs) in microbial communities in pipeline biofilms under antibiotic stress remains unexplored. In this study, the biodegradation efficiency of tetracycline (TCY) and sulfamethoxazole (SMX) was evaluated in a pilot reactor of the sewers. The results showed that under TCY and SMX stress, the degradation efficiency of sewage water was inhibited. The most abundant ARGs detected in the biofilm samples were TCY-related genes (e.g., tetW/N/W, tetC, and tetM), accounting for 34.1%. The microbial community composition varied, and the correlation analysis showed that antibiotic stress had a certain impact on the biological metabolic activity and function of the urban sewers. The community structure and diversity of biofilms enabled the evaluation of the bioconversion of antibiotics. Notably, Anaerocella and Paludibacter directly influenced the methanogenesis and sulfate reduction processes, playing a key role in the interaction between sulfate-reducing bacteria and methanogenic archaea. These microorganisms facilitated the proliferation of ARGs (tet and sul) in the biofilms through horizontal gene transfer. This study provides insight into the front-end control of ARGs, further improving sewage treatment plant processes and reducing the environmental and health risks caused by antibiotic abuse.202539894155
756490.9997Formation, 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
7561100.9997Deciphering 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
7901110.9997Responses of antibiotic resistance genes and microbial community in the microalgae-bacteria system under sulfadiazine: Mechanisms and implications. Microalgae-bacteria system is an emerging alternative for sustainable wastewater treatment. Exploring the structure and diversity of microbial community in microalgae-bacteria system under sulfadiazine stress can contribute to the understanding of the sulfadiazine behavior in environments. Furthermore, as important carriers of antibiotic resistance genes (ARGs), microalgae can influence the profiles of ARGs either directly or indirectly through the secretion of metabolites. However, the effects of sulfadiazine on ARGs dissemination of microalgae-bacteria systems remain underreported. Herein, the impacts of sulfadiazine (1 mg/L) on the structural diversity and metabolic activity of microorganisms were examined in microalgae-bacteria systems. Results showed that microalgae-bacteria system could remove NH(4)(+)-N better (about 72.3 %) than activated sludge system, and hydrolysis was the first step in sulfadiazine degradation. A high level of intI1 (5.7 × 10(4) copies/mL) was detected in the initial media of the microalgae-bacteria system. Microalgae could hamper the rate of horizontal gene transfer activation. Compared with activated sludge system, the abundance of sul genes (sul1, sul2, sul3, and sulA) was significantly lowered after treating with microalgae-bacteria system. Additionally, the number of proteins and the sum of polysaccharides in the extracellular polymeric substances of the activated sludge system were lower than those of the microalgae-bacteria system. Microalgae can alter microbial communities. The genus Rozellomycota predominated all samples. Fungi with relatively high abundance increased in the microalgae-bacteria system, including Dipodascaceae, Rhodotorula, and Geotrichum. These results offer valuable insights into the application processes involving microalgae-bacteria system.202540602895
8085120.9997Elevated CO(2) alleviated the dissemination of antibiotic resistance genes in sulfadiazine-contaminated soil: A free-air CO(2) enrichment study. Climate change affects soil microbial communities and their genetic exchange, and subsequently modifies the transfer of antibiotic resistance genes (ARGs) among bacteria. However, how elevated CO(2) impacts soil antibiotic resistome remains poorly characterized. Here, a free-air CO(2) enrichment system was used in the field to investigate the responses of ARGs profiles and bacterial communities to elevated CO(2) (+200 ppm) in soils amended with sulfadiazine (SDZ) at 0, 0.5 and 5 mg kg(-1). Results showed that SDZ exposure induced the co-occurrence of beta-lactamase and tetracycline resistance genes, and SDZ at 5 mg kg(-1) enhanced the abundance of aminoglycoside, sulfonamide and multidrug resistance genes. However, elevated CO(2) weakened the effects of SDZ at 0.5 mg kg(-1) following an observed reduction in the total abundance of ARGs and mobile genetic elements. Additionally, elevated CO(2) significantly decreased the abundance of vancomycin resistance genes and alleviated the stimulation of SDZ on the dissemination of aminoglycoside resistance genes. Correlation analysis and structural equation models revealed that elevated CO(2) could directly influence the spread of ARGs or impose indirect effects on ARGs by affecting soil properties and bacterial communities. Overall, our results furthered the knowledge of the dissemination risks of ARGs under future climate scenarios.202336857828
6965130.9997Metaphire guillelmi gut as hospitable micro-environment for the potential transmission of antibiotic resistance genes. Earthworm gut played an important role in the transformation of various contaminants in the soil environments. With the increasing application of organic fertilizer recently, the ingestion of antibiotics, antibiotic resistance bacteria (ARB), and antibiotic resistance genes (ARGs) made the earthworm gut a potential favorable micro-environment for the transmission of ARGs in the soil. In this work, the conventional plate incubation and high-throughput sequencing methods were both employed to investigate the composition of the cultivable and overall ARB/ARGs in the Metaphire guillelmi earthworm gut. A total of 87 cultivable isolates that resisted tetracycline (TC) and/or sulfadiazine (SD) were obtained, most of which belonged to phylum Firmicutes, genus Bacillus. Meanwhile, the counts of isolates with TC-SD dual resistance were higher than those with sole SD or TC resistance. Moreover, higher ARB counts and diversity were detected in the earthworm gut by high-throughput sequencing technique than those by the classical plate cultivation. Overall, the combination of conventional cultivable bacteria isolation and high-throughput sequencing methods provided a comprehensive understanding of the ARB composition in the earthworm gut. The results demonstrate that the earthworm gut is a hospitable micro-environment for ARB colonization. The potential role of earthworm intestinal ARB and ARGs proliferation in soil environments warrants further research.201930884260
7542140.9997Antibiotics resistance removal from piggery wastewater by an integrated anaerobic-aerobic biofilm reactor: Efficiency and mechanism. Antibiotic resistance residual in piggery wastewater poses serious threat to environment and human health. Biological treatment process is commonly installed to remove nutrient from piggery wastewater and also effective in removing antibiotics to varying degrees. But the specific pathways and mechanisms involved in the removal of antibiotic resistance are not yet well-understood. An integrated anaerobic-aerobic biofilm reactor (IAOBR) has been demonstrated efficient in removing conventional nutrients. It is here shown that the IAOBR effectively removed 79.0% of Sulfonamides, 55.7% of Tetracyclines and 53.6% of Quinones. Antibiotic resistance bacteria (ARB) were simultaneously inactivated by ~0.5 logs. Antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) were decreased by 0.51 logs and 0.42 logs, respectively. The antibiotics were mainly removed through aerobic compartments of the IAOBR. The mass loss of antibiotics in the reactor was achieved by biodegradation and adsorption, accounting for 52.1% and 47.9%, respectively. An obvious accumulation of ARGs was observed in the activated sludge. The potential host of ARGs was analyzed via microbial community and network. Partial least squares-structural equation model and correlation analysis revealed that the enrichment of ARGs was positively affected by MGEs, followed by bacterial community and ARBs, but the effect of antibiotics on ARGs was negative. Outcomes of this study provide valuable insights into the mechanisms of antibiotic resistance removal in biological treatment processes.202337714352
7022150.9997Fate 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
7552160.9997Effect 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
8083170.9997Alteration 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
8078180.9997Responses of bacterial communities and resistance genes on microplastics to antibiotics and heavy metals in sewage environment. In present study, copper (Cu), zinc (Zn), tetracycline (TC) and ampicillin (AMP) were selected to study the individual and synergistic effects of antibiotics and heavy metals on the microbial communities and resistance genes on polyvinyl chloride microplastics (PVC MPs) and surrounding sewage after 28 and 84 days. The results indicated that PVC MPs enriched many microorganisms from surrounding sewage, especially pathogenic bacteria such as Mycobacterium and Aquabacterium. The resistance gene with the highest abundance enriched on PVC MPs was tnpA (average abundance of 1.0 × 10(7) copies/mL sewage). The single presence of Zn, TC and AMP inhibited these enrichments for a short period of time (28 days). But the single presence of Cu and the co-existence of antibiotics and heavy metals inhibited these enrichments for a long period of time (84 days), resulting in relatively low microbial diversities and resistance genes abundances. Transpose tnpA had significantly positive correlations (p < 0.05) with all other genes. Pathogenic bacteria Mycobacterium and Legionella were potential hosts harboring 5 and 1 resistance genes, respectively. Overall, PVC MPs played important roles in the distribution and transfer of pathogenic bacteria and resistance genes in sewage with the presence of antibiotics or (and) heavy metals.202133254740
8077190.9997Effects of coexistence of tetracycline, copper and microplastics on the fate of antibiotic resistance genes in manured soil. The coexistence of antibiotics, heavy metals and microplastics is becoming commonplace and may affect antibiotic resistance in manured soil. The current understanding of the role of microplastics in soil with combined pollution of antibiotics, heavy metals and antibiotic resistance genes (ARGs) is limited. Here, the effects of the coexistence of tetracycline (TC), Cu and environmental microplastics (EM) on the fate of nine ARGs and three heavy metal resistance genes in agricultural soil were investigated by batch and microcosm experiments. EM were obtained by exposing virgin microplastics to soil environments for 80 days, which exhibited higher adsorption affinity for Cu and TC than soil particles and virgin microplastics. 1% EM in soil increased bioavailable concentrations of TC and Cu by 79-138% and 88-135%, respectively, and decreased TC dissipation from 11.79 mg kg(-1) to 3.08 mg kg(-1). Correspondingly, the total relative abundances of target ARGs increased by 219-348%. The significant correlations of tetG, tetB, tetQ, sul2, sul1 and intl1 with bioavailable fractions of TC and Cu in soil environments were revealed by network analysis. Moreover, scanning electron micrographs showed the special plastisphere around EM. Attributed to the biofilm generation and higher pollutant accumulation in the plastisphere, EM could be the source of antibiotic-resistant bacteria and ARGs in soil environments. Structure equation models further identified that indirect effects of EM acted a major role in the propagation of ARGs by altering soil properties, soil microbial diversity and intl1 abundance. This study revealed that EM could increase the stimulative effects of Cu and TC on antibiotic resistance and magnify the environmental risk of manure application in soil environments.202134091329