Microalgae Enhances the Adaptability of Epiphytic Bacteria to Sulfamethoxazole Stress and Proliferation of Antibiotic Resistance Genes Mediated by Integron. - Related Documents




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756501.0000Microalgae Enhances the Adaptability of Epiphytic Bacteria to Sulfamethoxazole Stress and Proliferation of Antibiotic Resistance Genes Mediated by Integron. The transmission of ARGs in the microalgae-associated epiphytic bacteria remains unclear under antibiotic exposure, apart from altering the microbial community structure. In this study, Chlorella vulgaris cocultured with bacteria screened from surface water was examined to explore the spread of ARGs in the presence of sulfamethoxazole (SMX). The extracellular polymers released by Chlorella vulgaris could reduce antibiotic-induced collateral damage to bacteria, thus increasing the diversity of the microalgae-associated epiphytic bacteria. The abundances of sul1 and intI1 in the phycosphere at 1 mg/L SMX dose increased by 290 and 28 times, respectively. Metagenomic sequencing further confirmed that SMX bioaccumulation stimulated the horizontal transfer of sul1 mediated by intI1 in the microalgae-associated epiphytic bacteria, while reactive oxygen species (ROS)-mediated oxidative stress induced the SOS response and thus enhanced the transformation of sul1 in the J group. This is the first study to verify that microalgae protect bacteria from antibiotic damage and hinder the spread of ARGs mediated by SOS response, while the transfer of ARGs mediated by integron is promoted due to the bioaccumulation of SMX in the phycosphere. The results contribute to present comprehensive understanding of the risk of ARG proliferation by the presence of emerging contaminants residues in river.202439417646
756610.9999Antibiotic sulfadiazine degradation by persulfate oxidation: Intermediates dependence of ecotoxicity and the induction of antibiotic resistance genes. To preserve the water resources, this study has analyzed the ecotoxicity and antibiotic resistance genes (ARGs) induction capacity of sulfadiazine degradation intermediates resulting from persulfate activation oxidation enhanced by ultraviolet, ultrasound and microwave. The five degradation pathways caused by the contribution discrepancy of electron transfer and singlet oxygen ((1)O(2)) and variations in the ecotoxicity of different degradation products were analyzed. Microcosm experiment exhibited that the microbial community in actual water changed significantly with SDZ and degradation intermediates, in which the dominant genera were Aeromonas, Cupriavidus, Elizabethkingia and Achromobacter. Except for the selective pressure on bacteria, the degradation intermediates also exert a certain degree or even stronger induction on sulfonamide ARGs (sul4, sul1 and sul2) than SDZ. Furthermore, the potential hosts for sulfonamide ARGs were revealed by network analysis. These results provide a better understanding of antibiotics degradation mechanism and ARGs occurrence, which is useful for controlling the spread of ARGs.202336372382
756420.9999Formation, 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
756030.9998The 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
851540.9998In vitro assessment of the bacterial stress response and resistance evolution during multidrug-resistant bacterial invasion of the Xenopus tropicalis intestinal tract under typical stresses. The intestinal microbiome might be both a sink and source of resistance genes (RGs). To investigate the impact of environmental stress on the disturbance of exogenous multidrug-resistant bacteria (mARB) within the indigenous microbiome and proliferation of RGs, an intestinal conjugative system was established to simulate the invasion of mARB into the intestinal microbiota in vitro. Oxytetracycline (OTC) and heavy metals (Zn, Cu, Pb), commonly encountered in aquaculture, were selected as typical stresses for investigation. Adenosine 5'-triphosphate (ATP), hydroxyl radical (OH·(-)) and extracellular polymeric substance (EPS) were measured to investigate their influence on the acceptance of RGs by intestinal bacteria. The results showed that the transfer and diffusion of RGs under typical combined stressors were greater than those under a single stressor. Combined effect of OTC and heavy metals (Zn, Cu) significantly increased the activity and extracellular EPS content of bacteria in the intestinal conjugative system, increasing intI3 and RG abundance. OTC induced a notable inhibitory response in Citrobacter and exerted the proportion of Citrobacter and Carnobacterium in microbiota. The introduction of stressors stimulates the proliferation and dissemination of RGs within the intestinal environment. These results enhance our comprehension of the typical stresses effect on the RGs dispersal in the intestine.202438280323
790150.9998Responses 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
856460.9998Effects of functional modules and bacterial clusters response on transmission performance of antibiotic resistance genes under antibiotic stress during anaerobic digestion of livestock wastewater. The formation and transmission of antibiotic resistance genes (ARGs) have attracted increasing attention. It is unclear whether the internal mechanisms by which antibiotics affect horizontal gene transfer (HGT) of ARGs during anaerobic digestion (AD) were influenced by dose and type. We investigated the effects of two major antibiotics (oxytetracycline, OTC, and sulfamethoxazole, SMX) on ARGs during AD according to antibiotic concentration in livestock wastewater influent. The low-dose antibiotic (0.5 mg/L) increased ROS and SOS responses, promoting the formation of ARGs. Meanwhile, low-dose antibiotics could also promote the spread of ARGs by promoting pili, communication responses, and the type IV secretion system (T4SS). However, different types and doses of antibiotics would lead to changes in the above functional modules and then affect the enrichment of ARGs. With the increasing dose of SMX, the advantages of pili and communication responses would gradually change. In the OTC system, low-dose has the strongest promoting ability in both pili and communication responses. Similarly, an increase in the dose of SMX would change T4SS from facilitation to inhibition, while OTC completely inhibits T4SS. Microbial and network analysis also revealed that low-dose antibiotics were more favorable for the growth of host bacteria.202336063716
856270.9998Nitrogen and phosphorus limitations promoted bacterial nitrate metabolism and propagation of antibiotic resistome in the phycosphere of Auxenochlorella pyrenoidosa. Despite that nitrogen (N) and phosphorus (P) play critical roles in the lifecycle of microalgae, how N and P further affect the distribution of bacteria and antibiotic resistance genes (ARGs) in the phycosphere is still poorly understood. In this study, the effects of N and P on the distribution of ARGs in the phycosphere of Auxenochlorella pyrenoidosa were investigated. Results showed that the growth and chlorophyll synthesis of microalgae were inhibited when N or P was limited, regardless of the N/P ratios, but the extracellular polymeric substances content and nitrate assimilation efficiency were enhanced in contrast. Metagenomic sequencing revealed that N or P limitation resulted in the recruitment of specific bacteria that highly contribute to the nitrate metabolism in the phycosphere. Besides, N or P limitation promoted the propagation of phycosphere ARGs, primarily through horizontal gene transfer mediated by mobile genetic elements. The enrichment of specific bacteria induced by changes in the algal physiology also contributed to the ARGs proliferation under nutrient limitation. Our results demonstrated that the reduction of algal cells caused by nutrient limitation could promote the propagation of ARGs, which provides new insights into the occurrence and spread of ARGs in the phycosphere.202438367442
808280.9998Deciphering the toxic effects of antibiotics on denitrification: Process performance, microbial community and antibiotic resistance genes. The extensive application of antibiotics, and the occurrence and spread of antibiotic resistance genes (ARGs) shade health risks to human and animal. The long-term effects of sulfamethoxazole (SMX) and tetracycline (TC) on denitrification process were evaluated in this study, with the focus on nitrogen removal performance, microbial community and ARGs. Results showed that low-concentration SMX and TC (<0.2 mg L(-1)) initially caused a deterioration in nitrogen removal performance, while higher concentrations (0.4-20 mg L(-1)) of both antibiotics had no further inhibitory influences. The abundances of ARGs in both systems generally increased during the whole period, and most of them had significant correlations with intI1, especially efflux-pump genes. Castellaniella, which was the dominant genus under antibiotic pressure, might be potential resistant bacteria. These findings provide an insight into the toxic effects of different antibiotics on denitrification process, and guides future efforts to control antibiotics pollution in ecosystems.202032250829
851690.9998Graphene 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
8563100.9998Overlooked role of extracellular polymeric substances in antibiotic-resistance gene transfer within microalgae-bacteria system. Controlling the spread of antibiotic-resistance genes (ARGs) under antibiotic stress has become an increasingly urgent issue. Microalgae possess the capability to remove antibiotics while concurrently inhibiting ARGs. Microalgae-bacteria systems can produce significant quantities of extracellular polymeric substances (EPS). However, the roles of EPS in the spread of ARGs have not been sufficiently explored, resulting in an insufficient understanding of the contribution of each EPS component and a lack of analysis on the complex interactions between EPS and ARGs. This study systematically explored the overlooked role of EPS in the transmission of ARGs within microalgae-bacteria systems. The current results showed that the potential of the microalgae-bacteria system for treating antibiotic wastewater. The tightly bound-EPS (TB-EPS) can acquire the higher absolute abundances of ARGs compared with the loosely bound-EPS (LB-EPS). The correlation coefficient between polysaccharides and TB-EPS ARGs was higher than that between polysaccharides and LB-EPS ARGs. The gene patterns of LB-EPS closely clustered with those of TB-EPS, while intracellular ARG gene patterns differed from both TB-EPS and LB-EPS. Metagenomic analyses indicated that the relative abundances of sul1 and sul2 were considerably higher at the beginning stage compared to the end stage. The abundance of Achromobacter, increased by the end stage, aligning with its potential to produce exopolysaccharide. Additionally, the absolute abundance of genes encoding exopolysaccharides (nagB and galE) and conjugative transfer transcription regulator (traF), increased over time. These findings enhanced our comprehension of the significance of EPS on the fate of ARGs in microalgae-bacteria systems during the treatment of antibiotic-contaminated wastewater.202539879767
7581110.9998Enhanced 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
7582120.9998Anaerobic 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
7966130.9998How 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
7611140.9998Response of microorganisms in biofilm to sulfadiazine and ciprofloxacin in drinking water distribution systems. Effects of sulfadiazine and ciprofloxacin on microorganisms in biofilm of drinking water distribution systems (DWDSs) were studied. The results verified that the increases of 16S rRNA for total bacteria and bacterial genus Hyphomicrobium were related to the promotion of antibiotic resistance genes (ARGs) and class 1 integrons (int1) in DWDSs with sulfadiazine and ciprofloxacin. Moreover, the bacteria showed higher enzymatic activities in DWDSs with sulfadiazine and ciprofloxacin, which resulted in more production of extracellular polymeric substances (EPS). The higher contents of EPS proteins and secondary structure β-sheet promoted bacterial aggregation and adsorption onto surface of pipelines to form biofilm. EPS can serve as a barrier for the microorganisms in biofilm. Therefore, the biofilm bacterial communities shifted and the 16S rRNA for total bacteria increased in DWDSs with antibiotics, which also drove the ARGs promotion. Furthermore, the two antibiotics exhibited stronger combined effects than that caused by sulfadiazine and ciprofloxacin alone.201930471500
7980150.9998Effect of dissolved biochar on the transfer of antibiotic resistance genes between bacteria. The spread of antibiotic resistance genes (ARGs) is a global environmental issue. Dissolved biochar is more likely to contact bacteria in water, producing ecological risks. This study explored the effects of dissolved biochar on ARGs transfer in bacteria. Conjugative transfer efficiency was significantly different following treatment with different types of dissolved biochar. Typically, humic acid-like substance in dissolved biochar can significantly improve the transfer efficiency of ARGs between bacteria. When the concentration of dissolved biochar was ≤10 mg biochar/mL, humic acid-like substance substantially promoted ARGs transfer. An increase in dissolved biochar concentration weakened the ARGs transfer from humic acid-like substance. The inhibitory effects of small-molecule matters dominated, decreasing conjugative transfer frequency. At a concentration of 100 mg biochar/mL, the conjugative transfer efficiency of all treatments was lower than that of control. Compared with corn straw dissolved biochar, there were more transconjugants in pine sawdust dissolved biochar. Following treatment with 10 mg biochar/mL pine sawdust dissolved biochar, the number of transconjugants was at its maximum; approximately 7.3 folds higher than the control. We also explored mechanisms by which dissolved biochar impacts conjugative transfer. Due to the complex composition of dissolved biochar, its effects on the expression of conjugative transfer-related genes were also dynamic. This study investigates the ecological risk of biochar and guides its scientific application.202134274650
7978160.9998Metagenomic insight into the enrichment of antibiotic resistance genes in activated sludge upon exposure to nanoplastics. Activated sludge is an important reservoir for the co-occurring emerging contaminants including nanoplastics (NPs) and antibiotic resistance genes (ARGs). However, the impacts and potential mechanisms of NPs on the fate of ARGs in activated sludge are not fully understood. Herein, we used metagenomic approach to investigate the responses of ARGs, host bacteria, mobile genetic elements (MGEs), and functional genes to polystyrene (PS) NPs at environmentally relevant (0.5 mg/L) and high stress concentrations (50 mg/L) in activated sludge. The results showed that 0.5 and 50 mg/L PS NPs increased the relative abundance of ARGs in the activated sludge by 58.68% and 46.52%, respectively (p < 0.05). Host tracking analysis elucidated that the hosts of ARGs were significantly enriched by PS NPs (p < 0.05), with Proteobacteria being the predominant host bacteria. Additionally, the occurrence of new ARGs hosts and the enrichment of MGEs and functional genes (i.e., genes related to SOS response, cell membrane permeability, and secretion system, etc.) indicated that PS NPs promoted horizontal gene transfer (HGT) of ARGs. Finally, path modeling analysis revealed that the proliferation of ARGs caused by PS NPs was primarily attributed to the enhancement of HGT and the enrichment of host bacteria. Our findings contribute to a comprehensive understanding of the spread risk of ARGs in activated sludge under NPs pollution.202439510298
7502170.9998Differential dose-response patterns of intracellular and extracellular antibiotic resistance genes under sub-lethal antibiotic exposure. Although antibiotics are one of the most significant factors contributing to the propagation of antibiotic resistance genes (ARGs), studies on the dose-response relationship at sub-lethal concentrations of antibiotics remain scarce, despite their importance for assessing the risks of antibiotics in the environment. In this study, we constructed a series of microcosms to investigate the propagation of intracellular (iARGs) and extracellular (eARGs) ARGs in both water and biofilms when exposed to antibiotics at various concentrations (1-100 μg/L) and frequencies. Results showed that eARGs were more abundant than iARGs in water, while iARGs were the dominant ARGs form in biofilms. eARGs showed differentiated dose-response relationships from iARGs. The abundance of iARGs increased with the concentration of antibiotics as enhanced selective pressure overcame the metabolic burden of antibiotic-resistant bacteria carrying ARGs. However, the abundance of eARGs decreased with increasing antibiotic concentrations because less ARGs were secreted from bacterial hosts at higher concentrations (100 μg/L). Furthermore, combined exposure to two antibiotics (tetracycline & imipenem) showed a synergistic effect on the propagation of iARGs, but an antagonistic effect on the propagation of eARGs compared to exposure to a single antibiotic. When exposed to antibiotic at a fixed total dose, one-time dosing (1 time/10 d) favored the propagation of iARGs, while fractional dosing (5 times /10 d) favored the propagation of eARGs. This study sheds light on the propagation of antibiotic resistance in the environment and can help in assessing the risks associated with the use of antibiotics.202337257347
7613180.9998Effect of bio-electrochemical system on the fate and proliferation of chloramphenicol resistance genes during the treatment of chloramphenicol wastewater. Bioelectrochemical systems can effectively degrade antibiotics, but there is the need to better understand the fate of antibiotic resistance bacteria and antibiotic resistance genes during the bioelectrochemical degradation of antibiotics. In this study, a BES was developed as a platform to investigate the fate of chloramphenicol resistance bacteria (CRB) and the expression of chloramphenicol resistance genes (CRGs) under different operating conditions during chloramphenicol biodegradation. The results indicated that chloramphenicol was effectively removed and chloramphenicol removal efficiency could be improved under less chloramphenicol concentration and more negative cathode potential. Higher chloramphenicol concentration enhanced the enrichment of CRB and expression of CRGs. Furthermore, the abundances of CRB were enhanced under more negative cathode potential, the expression of CRGs under less negative cathode potential were induced. However, both the enrichment of CRB and expression of CRGs could be moderated under a medium cathode potential. This result could provide the scientific reference for research about the fate of antibiotic resistance genes in bioelectrochemical systems.201728390239
7981190.9998Dissolved biochar eliminates the effect of Cu(II) on the transfer of antibiotic resistance genes between bacteria. The proliferation of antibiotic resistance genes (ARGs) has posed significant risks to human and environmental health. Research has confirmed that Cu(II) could accelerate the conjugative transfer of ARGs between bacteria. This study found that adding dissolved biochar effectively weakened or eliminated the Cu(II)-facilitated efficient transfer of ARGs. The efficiency of conjugative transfer was promoted after treatment with Cu(II) (0.05 mg/L) or dissolved biochar at a pyrolysis temperature of 300 °C. When exposed to the combination of Cu(II) and dissolved biochar, the transfer frequency was significantly reduced; this occurred regardless of the Cu(II) concentration or pyrolysis temperature of dissolved biochar. In particular, when the Cu(II) concentration exceeded 0.5 mg/L, the transfer efficiency was entirely inhibited. Gene expression analysis indicated that different treatments affect transfer efficiency by regulating the expression of three global regulatory genes: korA, korB, and trbA. Among them, humic acid repressed the expression of these genes; however, Cu(II) formed complex with the humic acid-like components, gradually weakening the inhibitive effect of these components. The promotion of low molecule organic matters dominated, resulting in a dynamic decline in the transfer efficiency. This study provides a new environmental contaminant treatment approach to eliminate the heavy metal-facilitated transfer of ARGs between bacteria.202234583164