Ubiquitous nanocolloids suppress the conjugative transfer of plasmid-mediated antibiotic resistance in aqueous environment. - Related Documents




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678201.0000Ubiquitous nanocolloids suppress the conjugative transfer of plasmid-mediated antibiotic resistance in aqueous environment. Nanocolloids (Nc) are widespread in natural water environment, whereas the potential effects of Nc on dissemination of antibiotic resistance remain largely unknown. In this study, Nc collected from the Yellow River in Henan province was tested for its ability to influence the conjugative transfer of resistant plasmid in aqueous environment. The results revealed that the conjugative transfer of RP4 plasmid between Escherichia coli was down-regulated by 52%-91% upon exposure to 1-10 mg/L Nc and the reduction became constant when the dose became higher (20-200 mg/L). Despite the exposure of Nc activated the anti-oxidation and SOS response in bacteria through up-regulating genes involved in glutathione biosynthesis and DNA recombination, the inhibition on the synthesis and secretion of extracellular polysaccharide induced the prevention of cell-cell contact, leading to the reduction of plasmid transfer. This was evidenced by the decreased bacterial adhesion and lowered levels of genes and metabolites relevant to transmembrane transport and D-glucose phosphorylation, as clarified in phenotypic, transcriptomics and metabolomics analysis of E. coli. The significant down-regulation of glycolysis/gluconeogenesis and TCA cycle was associated with the shortage of ATP induced by Nc. The up-regulation of global regulatory genes (korA and trbA) and the reduction of plasmid genes (trfAp, trbBp, and traG) expression also contributed to the suppressed conjugation of RP4 plasmid. The obtained findings remind that the role of ubiquitous colloidal particles is nonnegligible when practically and comprehensively assessing the risk of antibiotic resistance in the environment.202438801878
678110.9997Antibiotic-resistance gene transfer in antibiotic-resistance bacteria under different light irradiation: Implications from oxidative stress and gene expression. Due to the significant public health risks, there is substantial scientific interest in the increasing abundance of antibiotic-resistance bacteria (ARB) and the spread of antibiotic-resistance genes (ARGs) in aquatic environments. To clearly understand the mechanism of ARG transfer, this study examined the conjugative transfer of genes encoding resistance to cephalosporin (bla(CTX)) and polymyxin (mcr-1) from two antibiotic-resistant donor strains, namely E. coli DH5α (CTX) and E. coli DH5α (MCR), and to a streptomycin-resistant receptor strain (E. coli C600 (Sm)). Conjugative transfer was specifically studied under different light irradiation conditions including visible light (VL), simulated sunlight (SS) and ultraviolet light (UV(254nm)). Results show that the conjugative transfer frequency was not affected by VL irradiation, while it was slightly improved (2-10 fold) by SS irradiation and extremely accelerated (up to 100 fold) by UV irradiation. Furthermore, this study also explored the link between ARG transfer and stress conditions. This was done by studying physiological and biochemical changes; oxidative stress response; and functional gene expression of co-cultured AR-E. coli strains under stress conditions. When correlated with the transfer frequency results, we found that VL irradiation did not affect the physiological and biochemical characteristics of the bacteria, or induce oxidative stress and gene expression. For SS irradiation, oxidative stress occurred slowly, with a slight increase in the expression of target genes in the bacterial cells. In contrast, UV irradiation, rapidly inactivated the bacteria, the degree of oxidative stress was very severe and the expression of the target genes was markedly up-regulated. Our study could provide new insight into the underlying mechanisms and links between accelerated conjugative transfer and oxidative stress, as well as the altered expression of genes relevant to conjugation and other stress responses in bacterial cells.201930465986
676120.9997Exposure to Al(2)O(3) nanoparticles facilitates conjugative transfer of antibiotic resistance genes from Escherichia coli to Streptomyces. The spread of antibiotic resistance genes (ARGs) has become a global environmental issue; it has been found that nanoparticles (NPs) can promote the transfer of ARGs between bacteria. However, it remains unclear whether NPs can affect this kind of conjugation in Streptomyces, which mainly conjugate with other bacteria via spores. In the present study, we demonstrated that Al(2)O(3) NPs significantly promote the conjugative transfer of ARGs from Escherichia coli (E. coli) ET12567 to Streptomyces coelicolor (S. coelicolor) M145 without the use of heat shock method. The number of transconjugants induced by Al(2)O(3) particles was associated with the size and concentration of Al(2)O(3) particles, exposure time, and the ratio of E. coli and spores. When nanoparticle size was 30 nm at a concentration of 10 mg/L, the conjugation efficiency reached a peak value of 182 cfu/10(8) spores, which was more than 60-fold higher than that of the control. Compared with nanomaterials, bulk particles exhibited no significant effect on conjugation efficiency. We also explored the mechanisms by which NPs promote conjugative transfer. After the addition of NPs, the intracellular ROS content increased and the expression of the classical porin gene ompC was stimulated. In addition, ROS enhanced the mRNA expression levels of conjugative genes by inhibiting global regulation genes. Meanwhile, expression of the conjugation-related gene intA was also stimulated, ultimately increasing the number of transconjugants. Our results indicated that Al(2)O(3) NPs significantly promoted the conjugative transfer of ARGs from bacteria to spores and aggravated the diffusion of resistance genes in the environment.201931561730
677730.9997Unveiling the role of uranium in enhancing the transformation of antibiotic resistance genes. Transformation represents one of the most important pathways for the horizontal transfer of antibiotic resistance genes (ARGs), which enables competent bacteria to acquire extracellular ARGs from the surrounding environment. Both heavy metals and irradiation have been demonstrated to influence the bacterial transformation process. However, the impact of ubiquitously occurring radioactive heavy metals on the transformation of ARGs remains largely unknown. Here, we showed that a representative radioactive nuclide, uranium (U), at environmental concentrations (0.005-5 mg/L), improved the transformation frequency of resistant plasmid pUC19 into Escherichia coli by 0.10-0.85-fold in a concentration-dependent manner. The enhanced ARGs transformation ability under U stress was demonstrated to be associated with reactive oxygen species (ROS) overproduction, membrane damage, and up-regulation of genes related to DNA uptake and recombination. Membrane permeability and ROS production were the predominant direct and indirect factors affecting transformation ability, respectively. Our findings provide valuable insight into the underlying mechanisms of the impacts of U on the ARGs transformation process and highlight concerns about the exacerbated spread of ARGs in radioactive heavy metal-contaminated ecosystems, especially in areas with nuclear activity or accidents.202439208634
898240.9997Ampicillin Exposure and Glutathione Deficiency Synergistically Promote Conjugative Transfer of Plasmid-Borne Antibiotic Resistance Genes. Plasmid-mediated conjugation is an important pathway for the spread of antibiotic resistance genes (ARGs), posing a significant risk to global public health. It has been reported that the conjugative transfer of ARGs could be enhanced by oxidative stress. Whether endogenous glutathione (GSH), a major non-protein thiol compound involved in cellular redox homeostasis, influences conjugative transfer is unknown. In this study, we show that the deletion of the GSH biosynthesis gene gshA and ampicillin exposure synergistically promoted the conjugative transfer of plasmid RP4 bearing multiple ARGs from the soil bacterium Enterobacter sp. CZ-1 to Escherichia coli S17-1λπ in co-culture experiments and to diverse soil bacteria belonging to eight phyla, including some potential human pathogens, in a soil incubation experiment. The deletion of gshA increased ROS generation and cell membrane permeability, and upregulated the expression of the genes involved in intracellular oxidative stress regulation, membrane permeability, plasmid replication, and the SOS response process, especially under ampicillin exposure. These results suggest that endogenous GSH is an important factor affecting the spread of plasmid-borne ARGs. Exposure to antibiotics and environmental stresses that cause a depletion of endogenous GSH in vivo are likely to increase the risk of ARG dissemination in the environment.202540346915
677150.9997Triclosan at environmental concentrations can enhance the spread of extracellular antibiotic resistance genes through transformation. The dissemination of antibiotic resistance mediated by horizontal transfer of antibiotic resistance genes (ARGs) is exacerbating the global antibiotic crisis. Currently, little is known about whether non-antibiotic, anti-microbial (NAAM) chemicals are associated with the dissemination of ARGs in the environment. In this study, we aimed to evaluate whether a ubiquitous NAAM chemical, triclosan (TCS), is able to promote the transformation of plasmid-borne antibiotic resistance genes (ARGs). By using the plasmid pUC19 carrying ampicillin resistance genes as the extracellular ARG and a model microorganism Escherichia coli DH5ɑ as the recipient, we found that TCS at environmentally detected concentrations (0.2 μg/L to 20 μg/L) significantly enhanced the transformation of plasmid-borne ARGs into E. coli DH5ɑ for up to 1.4-fold. The combination of phenotypic experiments, genome-wide RNA sequencing and proteomic analyses revealed that TCS exposure stimulated the reactive oxygen species (ROS) production for 1.3- to 1.5-fold, induced bacterial membrane damage and up-regulated the translation of outer membrane porin. Moreover, general secretion system Sec (1.4-fold), twin arginine translocation system Tat (1.2-fold) and type IV pilus secretion systems (2.5-fold) were enhanced by TCS, which might contribute to the DNA searching/capture by pilus. Together, TCS might increase the transformation frequency of ARGs into E. coli DH5ɑ by ROS over-production, damaging cell membrane barrier, mediating the pilus capture of plasmid and the translocation of plasmid via cell membrane channels. This study reports that TCS could accelerate the transformation of extracellular ARGs to competent bacteria at environmentally relevant concentrations. The findings advance our understanding of the fate of ARGs in ecosystems and call for risk assessments of NAAM chemicals on disseminating antibiotic resistance.202032019018
677660.9997Natural sphalerite nanoparticles can accelerate horizontal transfer of plasmid-mediated antibiotic-resistance genes. Minerals and microorganisms are integral parts of natural environments, and they inevitably interact. Antibiotic-resistance genes (ARGs) significantly threaten modern healthcare. However, the effects of natural minerals on ARG propagation in aquatic systems are not fully understood. The present work studied the effects of natural sphalerite (NS) nanoparticles on the horizontal transfer of ARGs from Escherichia coli DH5α (CTX) (donor) to E. coli C600 (Sm) (recipient), and from E. coli DH5α (MCR) (donor) to E. coli C600 (Sm), and their underlying mechanisms. NS particles (0.5-50 mg L(-1)) induced an NS-concentration-dependent increase in conjugative transfer frequency. The underlying mechanisms associated with the facilitated ARG transfer included the production of intracellular reactive oxygen species, the SOS response, changes in bacterial cell morphology, and alteration of mRNA levels of bacterial cell membrane protein-related genes and genes associated with conjugative ARG transfer. The information herein offers new mechanistic understanding of risks of bacterial resistance resulting from NS.202031999971
677570.9997Copper nanoparticles and copper ions promote horizontal transfer of plasmid-mediated multi-antibiotic resistance genes across bacterial genera. The spread of antibiotic resistance has become a major concern for public health. As emerging contaminants, various metallic nanoparticles (NPs) and ionic heavy metals have been ubiquitously detected in various environments. Although previous studies have indicated NPs and ionic heavy metals could exhibit co-selection effects for antibiotic resistance, little is known about whether and how they could promote antibiotic resistance spread via horizontal gene transfer across bacterial genera. This study, we report both CuO NPs and copper ions (Cu(2+)) could stimulate the conjugative transfer of multiple-drug resistance genes. When exposing bacteria to CuO NPs or Cu(2+) at environmental-relevant and sub-inhibitory concentrations (e.g., 1-100 μmol/L), conjugation frequencies of plasmid-encoded antibiotic resistance genes across genera (i.e., from Escherichia coli to Pseudomonas putida) were significantly enhanced (p < 0.05). The over-production of reactive oxygen species played a crucial role in promoting conjugative transfer. Genome-wide RNA and protein sequencing suggested expressional levels of genes and proteins related to oxidative stress, cell membrane permeability, and pilus generation were significantly up-regulated under CuO NPs and Cu(2+) exposure (p < 0.05). This study provides insights in the contributions of NPs and heavy metals on the spread of antibiotic resistance.201931158594
678080.9997Enhanced uptake of antibiotic resistance genes in the presence of nanoalumina. Nanomaterial pollution and the spread of antibiotic resistance genes (ARGs) are global public health and environmental concerns. Whether nanomaterials could aid the transfer of ARGs released from dead bacteria into live bacteria to cause spread of ARGs is still unknown. Here, we demonstrated that nano-Al2O3 could significantly promote plasmid-mediated ARGs transformation into Gram-negative Escherichia coli strains and into Gram-positive Staphylococcus aureus; however, bulk Al2O3 did not have this effect. Under suitable conditions, 7.4 × 10(6) transformants of E. coli and 2.9 × 10(5) transformants of S. aureus were obtained from 100 ng of a pBR322-based plasmid when bacteria were treated with nano-Al2O3. Nanoparticles concentrations, plasmid concentrations, bacterial concentrations, interaction time between the nanomaterial and bacterial cells and the vortexing time affected the transformation efficiency. We also explored the mechanisms underlying this phenomenon. Using fluorescence in situ hybridization and scanning electron microscopy, we found that nano-Al2O3 damaged the cell membrane to produce pores, through which plasmid could enter bacterial cells. Results from reactive oxygen species (ROS) assays, genome-wide expression microarray profiling and quantitative real-time polymerase chain reactions suggested that intracellular ROS damaged the cell membrane, and that an SOS response promoted plasmid transformation. Our results indicated the environmental and health risk resulting from nanomaterials helping sensitive bacteria to obtain antibiotic resistance.201626946995
677390.9997Regulation of intracellular process by two-component systems: Exploring the mechanism of plasmid-mediated conjugative transfer. Plasmid-mediated conjugative transfer facilitates the dissemination of antibiotic resistance, yet the comprehensive regulatory mechanisms governing this process remain elusive. Herein, we established pure bacteria and activated sludge conjugation system to investigate the regulatory mechanisms of conjugative transfer, leveraging metformin as an exogenous agent. Transcriptomic analysis unveiled that substantial upregulation of genes associated with the two-component system (e.g., AcrB/AcrA, EnvZ/Omp, and CpxA/CpxR) upon exposure to metformin. Furthermore, downstream regulators of the two-component system, including reactive oxygen species (ROS), cytoplasmic membrane permeability, and adenosine triphosphate (ATP) production, were enhanced by 1.7, 1.4 and 1.1 times, respectively, compared to the control group under 0.1 mg/L metformin exposure. Moreover, flow sorting and high-throughput sequencing revealed increased microbial community diversity among transconjugants in activated sludge systems. Notably, the antibacterial potential of human pathogenic bacteria (e.g., Bacteroides, Escherichia-Shigella, and Lactobacillus) was augmented, posing a potential threat to human health. Our findings shed light on the spread of antibiotic resistance bacteria and assess the ecological risks associated with plasmid-mediated conjugative transfer in wastewater treatment systems.202438838482
6778100.9997Bisphenol S Promotes the Transfer of Antibiotic Resistance Genes via Transformation. The antibiotic resistance crisis has seriously jeopardized public health and human safety. As one of the ways of horizontal transfer, transformation enables bacteria to acquire exogenous genes naturally. Bisphenol compounds are now widely used in plastics, food, and beverage packaging, and have become a new environmental pollutant. However, their potential relationship with the spread of antibiotic resistance genes (ARGs) in the environment remains largely unexplored. In this study, we aimed to assess whether the ubiquitous bisphenol S (BPS) could promote the transformation of plasmid-borne ARGs. Using plasmid pUC19 carrying the ampicillin resistance gene as an extracellular ARG and model microorganism E. coli DH5α as the recipient, we established a transformation system. Transformation assays revealed that environmentally relevant concentrations of BPS (0.1-10 μg/mL) markedly enhanced the transformation frequency of plasmid-borne ARGs into E. coli DH5α up to 2.02-fold. Fluorescent probes and transcript-level analyses suggest that BPS stimulated increased reactive oxygen species (ROS) production, activated the SOS response, induced membrane damage, and increased membrane fluidity, which weakened the barrier for plasmid transfer, allowing foreign DNA to be more easily absorbed. Moreover, BPS stimulates ATP supply by activating the tricarboxylic acid (TCA) cycle, which promotes flagellar motility and expands the search for foreign DNA. Overall, these findings provide important insight into the role of bisphenol compounds in facilitating the horizontal spread of ARGs and emphasize the need to monitor the residues of these environmental contaminants.202439337307
8524110.9997Tebuconazole exacerbates co-occurrence and horizontal transfer of antibiotic resistance genes. As one of the most widely used pesticides in the global fungicide market, tebuconazole has become heavily embedded in soil along with antibiotic resistance genes (ARGs). However, it remains unclear whether the selective pressure produced by tebuconazole affects ARGs and their horizontal transfer. In this experiment, we simulated a tebuconazole-contaminated soil ecosystem and observed changes in the abundance of ARGs and mobile genetic element (MGEs) due to tebuconazole exposure. We also established a plasmid RP4-mediated conjugative transfer system to investigate in depth the impact of tebuconazole on the horizontal transfer of ARGs and its mechanism of action. The results showed that under tebuconazole treatment at concentrations ranging from 0 to 10 mg/L, there was a gradual increase in the frequency of plasmid conjugative transfer, peaking at 10 mg/L which was 7.93 times higher than that of the control group, significantly promoting horizontal transfer of ARGs. Further analysis revealed that the conjugative transfer system under tebuconazole stress exhibited strong ability to form biofilm, and the conjugative transfer frequency ratio of biofilm to planktonic bacteria varied with the growth cycle of biofilm. Additionally, scanning electron microscopy and flow cytometry demonstrated increased cell membrane permeability in both donor and recipient bacteria under tebuconazole stress, accompanied by upregulation of ompA gene expression controlling cell membrane permeability. Furthermore, enzyme activity assays indicated significant increases in CAT, SOD activity, and GSH content in recipient bacteria under tebuconazole stress. Moreover, expression levels of transmembrane transporter gene trfAp as well as genes involved in oxidative stress and SOS response were found to be correlated with the frequency of plasmid conjugative transfer.202439277355
6783120.9996Mechanism of earthworm coelomic fluid inhibits multidrug-resistant bacteria and blocks resistance transmission. Antibiotic resistance is a growing global health crisis, especially the spread of multi-drug resistance. In this study, the inhibitory effects of earthworm coelomic fluid (ECF) on multidrug-resistant bacteria (MRB) were investigated during employing vermicomposting to treat excess sludge generated from wastewater treatment. The results demonstrated that the ECF was able to inhibit, even completely decompose the MRB. Notably, when the ECF concentration reached 1.0 mg/mL, the intracellular reactive oxygen species (ROS) level increased by 46.7 %, while cell viability decreased by 55.2 % compared to the control, demonstrating that ECF exerts strong antibacterial activity by inducing oxidative stress and disrupting cellular homeostasis. Furthermore, ECF effectively degraded the DNA of MRB, with removal rates of aphA, KanR, and tetA reaching 51.8 %, 42.3 %, and 35.0 %, respectively, indicating its ability to eliminate resistance genes and hinder their potential transfer. Additionally, the upregulation of genes involved in signaling, DNA replication and repair, and energy metabolism pathways suggests a systemic stress response in MRB, further supporting the broad-spectrum inhibitory effects of ECF on bacterial viability and resistance maintenance. Taken together, these findings may open a door to naturally and ecologically combat antibiotic resistance in pollutants control in wastewater treatment.202540706790
8508130.9996Phenolic compounds promote the horizontal transfer of antibiotic resistance genes in activated sludge. Phenolic compounds are common organic pollutants in wastewater. During the wastewater treatment process, these compounds may influence the microbial community structure and functions. However, the impact of the phenolic compounds in the wastewater treatment plants on the horizontal transfer of antibiotic resistance genes (ARGs) has not been well assessed. In this study, we investigated the horizontal transfer of ARGs under the stress of phenolic compounds. The results showed that in pure culture bacteria system, p-nitrophenol (PNP), p-aminophenol (PAP) and phenol (PhOH) (10-100 mg/L) can significantly increase the horizontal transfer frequency of ARGs by 2.2-4.6, 3.6-9.4 and 1.9-9.0 fold, respectively. And, the RP4 plasmid transfer from Escherichia coli HB101 (E. coli HB101) to the bacteria in activated sludge increased obviously under the stress of phenolic compounds. Further investigation revealed that the PNP and PhOH at the concentration of 10-100 mg/L increased the production of reactive oxygen species and the permeability of cell membrane in the donor and recipient, which could be the causes of horizontal transfer of RP4 plasmid. In addition, it was also found that PNP, PAP and PhOH stress inhibit the expression of the global regulatory genes korB and trbA in the RP4 plasmid, and increase the expression level of the traF gene, thereby promoting the conjugative transfer of the RP4 plasmid. Taken together, these results improved our understanding of the horizontal transfer of ARGs under the stress of phenolic compounds and provided basic information for management of the systems that treat wastewater containing phenolic compounds.202134392203
8607140.9996Different paths, same destination: Bisphenol A and its substitute induce the conjugative transfer of antibiotic resistance genes. Antibiotic resistance genes are primarily spread through horizontal gene transfer in aquatic environments. Bisphenols, which are widely used in industry, are pervasive contaminants in such environments. This study investigated how environmentally relevant concentrations of bisphenol A and its substitute (bisphenol S, Bisphenol AP and Bisphenol AF) affect the spread of antibiotic resistance genes among Escherichia coli. As a result, bisphenol A and its three substitutes were found to promote the RP4 plasmid-mediated conjugative transfer of antibiotic resistance genes with different promotive efficiency. Particularly, bisphenol A and bisphenol S were found to induce more than double the incidence of conjugation at 0.1 nmol/L concentration. They therefore were selected as model compounds to investigate the involved mechanisms. Surprisingly, both slightly inhibited bacterial activity, but there was no significant increase in cell death. Bisphenols exposure changed the polymeric substances excreted by the bacteria, increased the permeability of their cell membranes, induced the secretion of antioxidant enzymes and generated reactive oxygen species. They also affected the expression of genes related to conjugative transfer by upregulating replication and DNA transfer genes and downregulating global regulatory genes. It should be noted that gene expression levels were higher in the BPS-exposed group than in the BPA-exposed group. The synthesis of bacterial metabolites and functional components was also significantly affected by bisphenols exposure. This research has helped to clarify the potential health risks of bisphenol contamination of aquatic environments.202439510271
6770150.9996Triclosan at environmentally relevant concentrations promotes horizontal transfer of multidrug resistance genes within and across bacterial genera. BACKGROUND: Antibiotic resistance poses an increasing threat to public health. Horizontal gene transfer (HGT) promoted by antibiotics is recognized as a significant pathway to disseminate antibiotic resistance genes (ARGs). However, it is unclear whether non-antibiotic, anti-microbial (NAAM) chemicals can directly promote HGT of ARGs in the environment. OBJECTIVES: We aimed to investigate whether triclosan (TCS), a widely-used NAAM chemical in personal care products, is able to stimulate the conjugative transfer of antibiotic multi-resistance genes carried by plasmid within and across bacterial genera. METHODS: We established two model mating systems, to investigate intra-genera transfer and inter-genera transfer. Escherichia coli K-12 LE392 carrying IncP-α plasmid RP4 was used as the donor, and E. coli K-12 MG1655 or Pseudomonas putida KT2440 were the intra- and inter-genera recipients, respectively. The mechanisms of the HGT promoted by TCS were unveiled by detecting oxidative stress and cell membrane permeability, in combination with Nanopore sequencing, genome-wide RNA sequencing and proteomic analyses. RESULTS: Exposure of the bacteria to environmentally relevant concentrations of TCS (from 0.02 μg/L to 20 μg/L) significantly stimulated the conjugative transfer of plasmid-encoded multi-resistance genes within and across genera. The TCS exposure promoted ROS generation and damaged bacterial membrane, and caused increased expression of the SOS response regulatory genes umuC, dinB and dinD in the donor. In addition, higher expression levels of ATP synthesis encoding genes in E. coli and P. putida were found with increased TCS dosage. CONCLUSIONS: TCS could enhance the conjugative ARGs transfer between bacteria by triggering ROS overproduction at environmentally relevant concentrations. These findings improve our awareness of the hidden risks of NAAM chemicals on the spread of antibiotic resistance.201830389380
8506160.9996Extracellular Polymeric Substances Acting as a Permeable Barrier Hinder the Lateral Transfer of Antibiotic Resistance Genes. Antibiotic resistance genes (ARGs) in bacteria are emerging contaminants as their proliferation in the environment poses significant threats to human health. It is well recognized that extracellular polymeric substances (EPS) can protect microorganisms against stress or damage from exogenous contaminants. However, it is not clear whether EPS could affect the lateral transfer of ARGs into bacteria, which is one of the major processes for the dissemination of ARGs. This study investigated the lateral transfer of ARGs carried by plasmids (pUC19, pHSG298, and pHSG396) into competent Escherichia coli cells with and without EPS. Transformant numbers and transformation efficiency for E. coli without EPS were up to 29 times of those with EPS at pH 7.0 in an aqueous system. The EPS removal further increased cell permeability in addition to the enhanced cell permeability by Ca(2+), which could be responsible for the enhanced lateral transfer of ARGs. The fluorescence quenching experiments showed that EPS could strongly bind to plasmid DNA in the presence of Ca(2+) and the binding strength (LogK (A) = 10.65-15.80 L mol(-1)) between EPS and plasmids was positively correlated with the enhancement percentage of transformation efficiency resulting from the EPS removal. X-ray photoelectron spectroscopy (XPS) analyses and model computation further showed that Ca(2+) could electrostatically bind with EPS mainly through the carboxyl group, hydroxyl group, and RC-O-CR in glucoside, thus bridging the plasmid and EPS. As a result, the binding of plasmids with EPS hindered the lateral transfer of plasmid-borne ARGs. This study improved our understanding on the function of EPS in controlling the fate and transport of ARGs on the molecular and cellular scales.201931057498
8981170.9996Response mechanisms of different antibiotic-resistant bacteria with different resistance action targets to the stress from photocatalytic oxidation. The stress response of antibiotic-resistant bacteria (ARB) and the spread of antibiotic resistance genes (ARGs) pose a serious threat to the aquatic environment and human beings. This study mainly explored the effect of the heterogeneous photocatalytic oxidation (UVA-TiO(2) system) on the stress response mechanism of ARB with different antibiotic resistance action targets, including the cell wall, proteins, DNA, RNA, folate and the cell membrane. Results indicate that the stress response mechanism of tetracycline- and sulfamethoxazole-resistant E. coli DH5α, which targets the synthesis of protein and folate, could rapidly induce global regulators by the overexpression of relative antibiotic resistance action target genes. Different stress response systems were mediated via cross-protection mechanism, causing stronger tolerance to an adverse environment than other ARB. Moreover, the photocatalytic inactivation mechanism of bacterial cells and a graded response of cellular stress mechanism caused differences in the intensity of the stress mechanism of antibiotic resistance action targets. E. coli DH5α resistant to cefotaxime and polymyxin, targeting synthesis of the cell wall and cell membrane, respectively, could confer greater advantages to bacterial survival and higher conjugative transfer frequency than E. coli DH5α resistant to nalidixic acid and rifampicin, which target the synthesis of DNA and RNA, respectively. This new perspective provides detailed information on the practical application of photocatalytic oxidation for inactivating ARB and hampering the spreading of ARGs in the aquatic environment.202235453030
8527180.9996Insight into the impacts and mechanisms of ketone stress on the antibiotic resistance in Escherichia coli. Accumulation of toxic organic has posed a substantial pressure on the proliferation of bacterial resistance. While aromatic organics have been demonstrated to enhance the antibiotic resistance in bacteria, no information is yet available on the effects of non-aromatic organics on the variations of bacterial resistance. Here, we investigated the effects of a typical ketone (i.e., methylisobutanone (MIBK)) on the variations of antibiotic resistance in Escherichia coli (E. coli). The results showed that the growth of resistant E. coli under environmental concentration of 50 μg/L MIBK was firstly inhibited as explained by the transient disruption in the cell membrane and then recovered possibly due to the reactive oxygen species. Exposure to 50 μg/L MIBK gradually raised the abundance of representative resistance gene (ampR) in E. coli. In contrast, the high concentration of 50 mg/L MIBK continuously inhibited the growth of resistant E. coli by disrupting cell membrane and notably promoted the proliferation of ampR through enhancing the horizontal transformation and up-regulating the expression of efflux pump gene. These findings provided the first evidence for the evolution of bacterial resistance in response to ketone organics.202235771331
8519190.9996Effects of Antibiotic Resistance Genes and Antibiotics on the Transport and Deposition Behaviors of Bacteria in Porous Media. Antibiotics present in the natural environment would induce the generation of antibiotic-resistant bacteria (ARB), causing great environmental risks. The effects of antibiotic resistance genes (ARGs) and antibiotics on bacterial transport/deposition in porous media yet are unclear. By using E. coli without ARGs as antibiotic-susceptible bacteria (ASB) and their corresponding isogenic mutants with ARGs in plasmids as ARB, the effects of ARGs and antibiotics on bacterial transport in porous media were examined under different conditions (1-4 m/d flow rates and 5-100 mM NaCl solutions). The transport behaviors of ARB were comparable with those of ASB under antibiotic-free conditions, indicating that ARGs present within cells had negligible influence on bacterial transport in antibiotic-free solutions. Interestingly, antibiotics (5-1000 μg/L gentamicin) present in solutions increased the transport of both ARB and ASB with more significant enhancement for ASB. This changed bacterial transport induced by antibiotics held true in solution with humic acid, in river water and groundwater samples. Antibiotics enhanced the transport of ARB and ASB in porous media via different mechanisms (ARB: competition of deposition sites; ASB: enhanced motility and chemotaxis effects). Clearly, since ASB are likely to escape sites containing antibiotics, these locations are more likely to accumulate ARB and their environmental risks would increase.202337406198