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247400.9806Nano-biosensors for rapid detection of antibiotic resistance genes blaCTX-M in Escherichia coli and blaKPC in Klebsiella pneumoniae. The wide spread of antibiotic resistance genes in clinical isolates of Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) has become a worrying issue worldwide. Thus, electrochemical DNA-based biosensors were developed and applied for the rapid detection of antibiotic resistance genes (blaCTX-M extended-spectrum β-lactamase (ESBL) in E. coli and blaKPC carbapenemase in Klebsiella pneumoniae). Selective ssDNA probes specific to the targeting complementary genes were designed and individually immobilized onto disposable sensor chips that were functionalized with nanocomposite (ssDNA-probe@ATP@AuNPs-PPY-V(2)O(5)). In this regard, the sensor surfaces were first modified with vanadium oxide (V(2)O(5)) and polypyrrole (PPy) then gold nanoparticles (AuNPs) to effectively promote the chemical immobilization of each of the designed bio-receptor (ssDNA probe) onto the sensor matrix using the 4-aminothiophenol (ATP) as the chemical cross-linker. Different parameters that affect the biosensing performance were studied and optimized. Accordingly, high sensitivity to both E. coli blaCTX-M and K. pneumoniae blaKPC genes were obtained with a dynamic linear range from 10(-6) to 0.1 ng/μL with correlation coefficient (R(2)) of 0.991 and 0.988, limit of detection (LODs) of 0.5 × 10(-7) ng/μL and 1 × 10(-7) ng/μL, and limit of quantification (LOQs) of 1.2 × 10(-7) and 3 × 10(-7) ng/μL, respectively. High selectivity was provided since no electrochemical signals were obtained from various competitors of common bacteria. Finally, different types of clinical samples were analyzed and validated.202540250654
782810.9799Simultaneous elimination of antibiotic-resistant bacteria and antibiotic resistance genes by different Fe-N co-doped biochars activating peroxymonosulfate: The key role of pyridine-N and Fe-N sites. The coexistence of antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB) in the environment poses a potential threat to public health. In our study, we have developed a novel advanced oxidation process for simultaneously removing ARGs and ARB by two types of iron and nitrogen-doped biochar derived from rice straw (FeN-RBC) and sludge (FeN-SBC). All viable ARB (approximately 10(8) CFU mL(-1)) was inactivated in the FeN-RBC/ peroxymonosulfate (PMS) system within 40 min and did not regrow after 48 h even in real water samples. Flow cytometry identified 96.7 % of dead cells in the FeN-RBC/PMS system, which verified the complete inactivation of ARB. Thorough disinfection of ARB was associated with the disruption of cell membranes and intracellular enzymes related to the antioxidant system. Whereas live bacteria (approximately 200 CFU mL(-1)) remained after FeN-SBC/PMS treatment. Intracellular and extracellular ARGs (tetA and tetB) were efficiently degraded in the FeN-RBC/PMS system. The production of active species, primarily •OH, SO(4)(•-) and Fe (IV), as well as electron transfer, were essential to the effective disinfection of FeN-RBC/PMS. In comparison with FeN-SBC, the better catalytic performance of FeN-RBC was mainly ascribed to its higher amount of pyridine-N and Fe(0), and more reactive active sites (such as CO group and Fe-N sites). Density functional theory calculations indicated the greater adsorption energy and Bader charge, more stable Fe-O bond, more easily broken OO bond in FeN-RBC/PMS, which demonstrated the stronger electron transfer capacity between FeN-RBC and PMS. To encapsulate, our study provided an efficient and dependable method for the simultaneous elimination of ARGs and ARB in water.202438669989
507020.9794Sequence-specific DNA solid-phase extraction in an on-chip monolith: Towards detection of antibiotic resistance genes. Antibiotic resistance of bacteria is a growing problem and presents a challenge for prompt treatment in patients with sepsis. Currently used methods rely on culturing or amplification; however, these steps are either time consuming or suffer from interference issues. A microfluidic device was made from black polypropylene, with a monolithic column modified with a capture oligonucleotide for sequence selective solid-phase extraction of a complementary target from a lysate sample. Porous properties of the monolith allow flow and hybridization of a target complementary to the probe immobilized on the column surface. Good flow-through properties enable extraction of a 100μL sample and elution of target DNA in 12min total time. Using a fluorescently labeled target oligonucleotide related to Verona Integron-Mediated Metallo-β-lactamase it was possible to extract and detect a 1pM sample with 83% recovery. Temperature-mediated elution by heating above the duplex melting point provides a clean extract without any agents that interfere with base pairing, allowing various labeling methods or further downstream processing of the eluent. Further integration of this extraction module with a system for isolation and lysis of bacteria from blood, as well as combining with single-molecule detection should allow rapid determination of antibiotic resistance.201728734608
508430.9791Cloth-based hybridization array system for the identification of antibiotic resistance genes in Salmonella. A simple macroarray system based on the use of polyester cloth as the solid phase for DNA hybridization has been developed for the identification and characterization of bacteria on the basis of the presence of various virulence and toxin genes. In this approach, a multiplex polymerase chain reaction (PCR) incorporating digoxigenin-dUTP is used to simultaneously amplify different marker genes, with subsequent rapid detection of the amplicons by hybridization with an array of probes immobilized on polyester cloth and immunoenzymatic assay of the bound label. As an example of the applicability of this cloth-based hybridization array system (CHAS) in the characterization of foodborne pathogens, a method has been developed enabling the detection of antibiotic resistance and other marker genes associated with the multidrug-resistant food pathogen Salmonella enterica subsp. enterica serotype Typhimurium DT104. The CHAS is a simple, cost-effective tool for the simultaneous detection of amplicons generated in a multiplex PCR, and the concept is broadly applicable to the identification of key pathogen-specific marker genes in bacterial isolates.200718363231
782940.9790Insights into capture-inactivation/oxidation of antibiotic resistance bacteria and cell-free antibiotic resistance genes from waters using flexibly-functionalized microbubbles. The spread of antibiotic resistance in the aquatic environment severely threatens the public health and ecological security. This study investigated simultaneously capturing and inactivating/oxidizing the antibiotic resistant bacteria (ARB) and cell-free antibiotic resistance genes (ARGs) in waters by flexibly-functionalized microbubbles. The microbubbles were obtained by surface-modifying the bubbles with coagulant (named as coagulative colloidal gas aphrons, CCGAs) and further encapsulating ozone in the gas core (named as coagulative colloidal ozone aphrons, CCOAs). CCGAs removed 92.4-97.5% of the sulfamethoxazole-resistant bacteria in the presence of dissolved organic matter (DOM), and the log reduction of cell-free ARGs (particularly, those encoded in plasmid) reached 1.86-3.30. The ozone release from CCOAs led to efficient in-situ oxidation: 91.2% of ARB were membrane-damaged and inactivated. In the municipal wastewater matrix, the removal of ARB increased whilst that of cell-free ARGs decreased by CCGAs with the DOM content increasing. The ozone encapsulation into CCGAs reinforced the bubble performance. The predominant capture mechanism should be electrostatic attraction between bubbles and ARB (or cell-free ARGs), and DOM enhanced the sweeping and bridging effect. The functionalized microbubble technology can be a promising and effective barrier for ARB and cell-free ARGs with shortened retention time, lessened chemical doses and simplified treatment unit.202235063836
783450.9789Elimination of representative antibiotic-resistant bacteria, antibiotic resistance genes and ciprofloxacin from water via photoactivation of periodate using FeS(2). The propagation of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) induced by the release of antibiotics poses great threats to ecological safety and human health. In this study, periodate (PI)/FeS(2)/simulated sunlight (SSL) system was employed to remove representative ARB, ARGs and antibiotics in water. 1 × 10(7) CFU mL(-1) of gentamycin-resistant Escherichia coli was effectively disinfected below limit of detection in PI/FeS(2)/SSL system under different water matrix and in real water samples. Sulfadiazine-resistant Pseudomonas and Gram-positive Bacillus subtilis could also be efficiently sterilized. Theoretical calculation showed that (110) facet was the most reactive facet on FeS(2) to activate PI for the generation of reactive species (·OH, ·O(2)(-), h(+) and Fe(IV)=O) to damage cell membrane and intracellular enzyme defense system. Both intracellular and extracellular ARGs could be degraded and the expression levels of multidrug resistance-related genes were downregulated during the disinfection process. Thus, horizontal gene transfer (HGT) of ARB was inhibited. Moreover, PI/FeS(2)/SSL system could disinfect ARB in a continuous flow reactor and in an enlarged reactor under natural sunlight irradiation. PI/FeS(2)/SSL system could also effectively degrade the HGT-promoting antibiotic (ciprofloxacin) via hydroxylation and ring cleavage process. Overall, PI/FeS(2)/SSL exhibited great promise for the elimination of antibiotic resistance from water.202438917629
786860.9788A double-quenching paperclip ECL biosensing platform for ultrasensitive detection of antibiotic resistance genes (mecA) based on Ti(3)C(2) MXene-Au NPs as a coreactant accelerator. The global spread of environmental biological pollutants, such as antibiotic-resistant bacteria and their antibiotic resistance genes (ARGs), has emerged as a critical public health concern. It is imperative to address this pressing issue due to its potential implications for public health. Herein, a DNA paperclip probe with double-quenching function of target cyclic cleavage was proposed, and an electrochemiluminescence (ECL) biosensing platform was constructed using Ti(3)C(2) MXene in-situ reduction growth of Au NPs (TCM-Au) as a coreactant accelerator, and applied to the sensitive detection of ARGs. Thanks to the excellent catalytic performance, large surface area and Au-S affinity of TCM-Au, the ECL performance of CdS QDs have been significantly improved. By cleverly utilizing the negative charge of the paperclip nucleic acid probe and its modification group, double-quenching of the ECL signal was achieved. This innovative approach, combined with target cyclic amplification, facilitated specific and sensitive detection of the mecA gene. This biosensing platform manifested highly selective and sensitive determination of mecA genes in the range of 10 fM to 100 nM and a low detection limit of 2.7 fM. The credible detectability and anti-interference were demonstrated in Yangtze river and Aeration tank outlet, indicating its promising application toward pollution monitoring of ARGs.202337666010
781270.9788Using the heat generated from electrically conductive concrete slabs to reduce antibiotic resistance in beef cattle manure. Proper treatment is necessary to reduce antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in livestock manure before land application. Conventional stockpiling suffers unreliable removal efficiency, while composting can be complicated and expensive. The objective of this study was to test the feasibility of a novel heat-based technology, i.e., stockpiling manure on conductive concrete slabs, to inactivate ARB and ARGs in beef cattle manure. In this study, two independent bench-scale trials were conducted. In both trials, samples were taken from manure piles on conductive concrete slabs and regular slabs (i.e., heated and unheated piles). In the heated pile of the first trial, 25.9% and 83.5% of the pile volume met the EPA Class A and Class B biosolids standards, respectively. For the heated pile of the second trial, the two values were 43.9% and 74.2%. In both trials, nearly all forms of the total and resistant Escherichia coli and enterococci were significantly lower in the heated piles than in the unheated piles. Besides, significant reduction of ARGs in heated piles was observed in the first trial. Through this proof-of-concept study, the new technology based on conductive concrete slabs offers an alternative manure storage method to conventional stockpiling and composting with respect to reduce ARB and ARGs in manure.202133736325
356680.9786Transfer of antibiotic multiresistant plasmid RP4 from escherichia coli to activated sludge bacteria. In situ transfer of a self-transmissible, antibiotic-multiresistant plasmid RP4 from a laboratory Escherichia coli strain C600 to indigenous activated sludge bacteria was investigated using filter mating. The transfer frequency of RP4 from the donor E. coli to the bacteria that was sampled from two wastewater treatment plants was 5.1x10(-2) to 7.5x10(-1) and 4.6x10(-3) to 1.3x10(-2)/potential recipient. The isolated transconjugants showed resistance to Ap, Km, and Tc and the presence of a plasmid with a similar size to RP4. The traG gene on RP4 was also detected from all transconjugants. Reverse-transfer experiments from the transconjugants to E. coli HB101 indicated that RP4 maintained self-transmissibility in the transconjugants. The transconjugant strains were dominant bacteria in activated sludge including Pseudomonas fluorescens, P. putida, and Ochrobactrum anthropi and minor populations of enteric bacterial strains including Citrobacter freundii, E. coli, Enterobacter cloacae, E. asburiae, and Klebsiella pneumoniae ssp. pneumoniae. The transconjugant strains K. pneumoniae ssp. pneumonia, E. cloacae, and E. asburiae had several naturally occurring plasmids. These results suggest that in situ transfer of plasmids and the exchange of antibiotic-resistant genes can occur between released and indigenous bacteria in activated sludge.200818930008
843190.9786A quaternary ammonium salt grafted tannin-based flocculant boosts the conjugative transfer of plasmid-born antibiotic resistance genes: The nonnegligible side of their flocculation-sterilization properties. This study developed dual-function tannin-based flocculants, namely tannin-graft-acrylamide-diallyl dimethyl ammonium chloride (TGCC-A/TGCC-C), endowed with enhanced flocculation-sterilization properties. The impacts of these flocculants on proliferation and transformation of antibiotic resistance genes (ARGs) among bacteria during the flocculation-deposition process were examined. TGCC-A/TGCC-C exhibited remarkable flocculation capacities towards both Escherichia coli and Staphylococcus aureus, encompassing a logarithmic range of initial cell density (10(8)-10(9) CFU/mL) and a broad pH spectrum (pH 2-11). The grafted quaternary ammonium salt groups played pivotal parts in flocculation through charge neutralization and bridging mechanisms, concurrently contributing to sterilization by disrupting cellular membranes. The correlation between flocculation and sterilization entails a sequential progression, where an excess of TGCC, initially employed for flocculation, is subsequently consumed for sterilization purposes. The frequencies of ARGs conjugative transfer were enhanced in bacterial flocs across all TGCC treatments, stemming from augmented bacterial aggregation and cell membrane permeability, elicited stress response, and up-regulated genes encoding plasmid transfer. These findings underscore the indispensable role of flocculation-sterilization effects in mediating the propagation of ARGs, consequently providing substantial support for the scientific evaluation of the environmental risks associated with flocculants in the context of ARGs dissemination during the treatment of raw water featuring high bacterial density.202337619725
7858100.9785Photocatalytic Reactive Ultrafiltration Membrane for Removal of Antibiotic Resistant Bacteria and Antibiotic Resistance Genes from Wastewater Effluent. Biological wastewater treatment is not effective in removal of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). In this study, we fabricated a photocatalytic reactive membrane by functionalizing polyvinylidene fluoride (PVDF) ultrafiltration (UF) membrane with titanium oxide (TiO(2)) nanoparticles for the removal of ARB and ARGs from a secondary wastewater effluent. The TiO(2)-modified PVDF membrane provided complete retention of ARB and effective photocatalytic degradation of ARGs and integrons. Specifically, the total removal efficiency of ARGs (i.e., plasmid-mediated floR, sul1, and sul2) with TiO(2)-modified PVDF membrane reached ∼98% after exposure to UV irradiation. Photocatalytic degradation of ARGs located in the genome was found to be more efficient than those located in plasmid. Excellent removal of integrons (i.e., intI1, intI2, and intI3) after UV treatment indicated that the horizontal transfer potential of ARGs was effectively controlled by the TiO(2) photocatalytic reaction. We also evaluated the antifouling properties of the TiO(2)-UF membrane to demonstrate its potential application in wastewater treatment.201829984583
7786110.9785Effect of solar photo-Fenton process in raceway pond reactors at neutral pH on antibiotic resistance determinants in secondary treated urban wastewater. Solar photo-Fenton process in raceway pond reactors was investigated at neutral pH as a sustainable tertiary treatment of real urban wastewater. In particular, the effect on antibiotic resistance determinants was evaluated. An effective inactivation of different wild bacterial populations was achieved considering total and cefotaxime resistant bacteria. The detection limit (1 CFU mL(-1)) was achieved in the range 80-100 min (5.4-6.7 kJ L(-1) of cumulative solar energy required) for Total Coliforms (TC) (40-60 min for resistant TC, 4.3-5.2 kJ L(-1)), 60-80 min (4.5-5.4 kJ L(-1)) for Escherichia coli (E. coli) (40 min for resistant E. coli, 4.1-4.7 kJ L(-1)) and 40-60 min (3.9-4.5 kJ L(-1)) for Enterococcus sp. (Entero) (30-40 min for resistant Entero, 3.2-3.8 kJ L(-1)) with 20 mg L(-1) Fe(2+) and 50 mg L(-1) H(2)O(2). Under these mild oxidation conditions, 7 out of the 10 detected antibiotics were effectively removed (60-100%). As the removal of antibiotic resistance genes (ARGs) is of concern, no conclusive results were obtained, as sulfonamide resistance gene was reduced to some extent (relative abundance <1), meanwhile class 1 integron intI1 and ß-lactam resistance genes were not affected. Accordingly, more research and likely more intensive oxidative conditions are needed for an efficient ARGs removal.201931202058
7788120.9785Inactivation of antibiotic resistant Escherichia coli and degradation of its resistance genes by glow discharge plasma in an aqueous solution. Emerging contaminants such as antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) are becoming a global environmental problem. In this study, the glow discharge plasma (GDP) was applied for degrading antibiotic resistant Escherichia coli (E. coli) with resistance genes (tetA, tetR, aphA) and transposase gene (tnpA) in 0.9% sterile saline. The results showed that GDP was able to inactivate the antibiotic resistant E. coli and remove the ARGs and reduce the risk of gene transfer. The levels of E. coli determined by 16S rRNA decreased by approximately 4.7 logs with 15 min of discharge treatment. Propidium monoazide - quantitative polymerase chain reaction (PMA-qPCR) tests demonstrated that the cellular structure of 4.8 more logs E. coli was destroyed in 15 min. The reduction of tetA, tetR, aphA, tnpA genes was increased to 5.8, 5.4, 5.3 and 5.5 logs with 30 min discharge treatment, respectively. The removal of ARGs from high salinity wastewater was also investigated. The total abundance of ARGs was reduced by 3.9 logs in 30 min. Scavenging tests indicated that hydroxyl radicals (·OH) was the most probable agents for bacteria inactivation and ARGs degradation. In addition, the active chlorine (Cl· and Cl(2)) which formed during the discharge may also contribute to the inactivation and degradation.202032229364
6788130.9785Release and Constancy of an Antibiotic Resistance Gene in Seawater under Grazing Stress by Ciliates and Heterotrophic Nanoflagellates. Extracellular DNA (exDNA) is released from bacterial cells through various processes. The antibiotic resistance genes (ARGs) coded on exDNA may be horizontally transferred among bacterial communities by natural transformation. We quantitated the released/leaked tetracycline resistance gene, tet(M) over time under grazing stress by ciliates and heterotrophic nanoflagellates (HNFs), and found that extracellular tet(M) (ex-tetM) increased with bacterial grazing. Separate microcosms containing tet(M)-possessing bacteria with ciliates or HNFs were prepared. The copy number of ex-tetM in seawater in the ciliate microcosm rapidly increased until 3 d after the incubation, whereas that in the HNF microcosm showed a slower increase until 20 d. The copy number of ex-tetM was stable in both cases throughout the incubation period, suggesting that extracellular ARGs are preserved in the environment, even in the presence of grazers. Additionally, ARGs in bacterial cells were constant in the presence of grazers. These results suggest that ARGs are not rapidly extinguished in a marine environment under grazing stress.201728592722
7863140.9785Mechanisms on the removal of gram-negative/positive antibiotic resistant bacteria and inhibition of horizontal gene transfer by ferrate coupled with peroxydisulfate or peroxymonosulfate. The existence of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) has been a global public environment and health issue. Due to the different cell structures, gram-positive/negative ARB exhibit various inactivation mechanisms in water disinfection. In this study, a gram-negative ARB Escherichia coli DH5α (E. coli DH5α) was used as a horizontal gene transfer (HGT) donor, while a gram-positive ARB Bacillus as a recipient. To develop an efficient and engineering applicable method in water disinfection, ARB and ARGs removal efficiency of Fe(VI) coupled peroxydisulfate (PDS) or peroxymonosulfate (PMS) was compared, wherein hydroxylamine (HA) was added as a reducing agent. The results indicated that Fe(VI)/PMS/HA showed higher disinfection efficiency than Fe(VI)/PDS/HA. When the concentration of each Fe(VI), PMS, HA was 0.48 mM, 5.15 log E. coli DH5α and 3.57 log Bacillus lost cultivability, while the proportion of recovered cells was 0.0017 % and 0.0566 %, respectively, and HGT was blocked. Intracellular tetA was reduced by 2.49 log. Fe(IV) and/or Fe(V) were proved to be the decisive reactive species. Due to the superiority of low cost as well as high efficiency and practicality, Fe(VI)/PMS/HA has significant application potential in ARB, ARGs removal and HGT inhibition, offering a new insight for wastewater treatment.202438615644
7865150.9784Inactivation of antibiotic resistant bacteria by Fe(3)O(4) @MoS(2) activated persulfate and control of antibiotic resistance dissemination risk. Antibiotic resistance poses a global environmental challenge that jeopardizes human health and ecosystem stability. Antibiotic resistant bacteria (ARB) significantly promote the spreading and diffusion of antibiotic resistance. This study investigated the efficiency and mechanism of inactivating tetracycline-resistant Escherichia coli (TR E. coli) using Fe(3)O(4) @MoS(2) activated persulfate (Fe(3)O(4) @MoS(2)/PS). Under optimized conditions (200 mg/L Fe(3)O(4) @MoS(2), 4 mM PS, 35 °C), TR E. coli (∼7.5 log CFU/mL) could be fully inactivated within 20 min. The primary reactive oxygen species (ROS) responsible for TR E. coli inactivation in the Fe(3)O(4) @MoS(2)/PS system were hydroxyl radicals (•OH) and superoxide radicals (•O(2)(-)). Remarkably, the efflux pump protein was targeted and damaged by the generated ROS during the inactivation process, resulting in cell membrane rupture and efflux of cell content. Additionally, the horizontal transmission ability of residual antibiotic resistance genes (ARGs) harboring in the TR E. coli was also reduced after the inactivation treatment. This study offers an efficient approach for TR E. coli inactivation and substantial mitigation of antibiotic resistance dissemination risk.202438286046
7860160.9784Enhanced removal of antibiotic-resistant bacteria and resistance genes by three-dimensional electrochemical process using MgFe(2)O(4)-loaded biochar as both particle electrode and catalyst for peroxymonosulfate activation. In this study, MgFe(2)O(4)-loaded biochar (MFBC) was used as a three-dimensional particle electrode to active peroxymonosulfate (EC/MFBC/PMS) for the removal of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). The results demonstrated that, under the conditions of 1.0 mM PMS concentration, 0.4 g/L material dosage, 5 V voltage intensity, and MFBC preparation temperature of 600 °C, the EC/MFBC600/PMS system achieved complete inactivation of E. coli DH5α within 5 min and the intracellular sul1 was reduced by 81.5 % after 30 min of the treatment. Compared to EC and PMS alone treatments, the conjugation transfer frequency of sul1 rapidly declined by 92.9 % within 2 min. The cell membrane, proteins, lipids, as well as intracellular and extracellular ARGs in E. coli DH5α were severely damaged by free radicals in solution and intracellular reactive oxygen species (ROS). Furthermore, up-regulation was observed in genes associated with oxidative stress, SOS response and cell membrane permeability in E. coli DH5α, however, no significant changes were observed in functional genes related to gene conjugation and transfer mechanisms. This study would contribute to the underlying of PMS activation by three-dimensional particle electrode, and provide novel insights into the mechanism of ARB inactivation and ARGs degradation under PMS advanced oxidation treatment.202439197284
9995170.9783Direct fluorescence in situ hybridization (FISH) in Escherichia coli with a target-specific quantum dot-based molecular beacon. Quantum dots (QDs) are inorganic fluorescent nanocrystals with excellent properties such as tunable emission spectra and photo-bleaching resistance compared with organic dyes, which make them appropriate for applications in molecular beacons. In this work, quantum dot-based molecular beacons (QD-based MBs) were fabricated to specifically detect β-lactamase genes located in pUC18 which were responsible for antibiotic resistance in bacteria Escherichia coli (E. coli) DH5α. QD-based MBs were constructed by conjugating mercaptoacetic acid-quantum dots (MAA-QDs) with black hole quencher 2 (BHQ2) labeled thiol DNA vial metal-thiol bonds. Two types of molecular beacons, double-strands beacons and hairpin beacons, were observed in product characterization by gel electrophoresis. Using QD-based MBs, one-step FISH in tiny bacteria DH5α was realized for the first time. QD-based MBs retained their bioactivity when hybridizing with complementary target DNA, which showed excellent advantages of eliminating background noise caused by adsorption of non-specific bioprobes and achieving clearer focus of genes in plasmids pUC18, and capability of bacterial cell penetration and signal specificity in one-step in situ hybridization.201020729070
7805180.9783Elimination of antibiotic-resistance bacterium and its associated/dissociative bla(TEM-1) and aac(3)-II antibiotic-resistance genes in aqueous system via photoelectrocatalytic process. The ubiquity of antibiotic-resistance bacteria (ARB) and antibiotic-resistance genes (ARGs) in various environmental matrices is a potential threat to human and ecological health. Therefore, the inactivation of ARB E. coli S1-23 and the elimination of its associated ARGs, bla(TEM-1) and aac(3)-II, were investigated using the photoelectrocatalytic (PEC) process. Results indicate that the ARB E. coli S1-23 (1 × 10(8) cfu mL(-1)) and its ARGs (extracellular and intracellular) could be fully inactivated within 10 and 16 h PEC treatment, respectively. In contrast, photocatalytic (PC) and electrochemical (EC) treatments displayed no obvious effect; however, ARG-containing DNA extracted from E. coli S1-23, which was used as a model for dissociative naked ARGs, could be completely decomposed within a few minutes through these three treatments. Further analyses, including PCR, AFM and HPLC, proved that the structural integrity and surface topography of naked ARGs are damaged during treatment and can be completely eliminated. Furthermore, there is no generation of cytosine, guanine, adenine or thymine intermediates during the PEC, PC, and EC treatments. This study is the first report to propose the PEC treatment as a promising method for complete decomposition of ARB and ARGs in aqueous systems.201728863344
7846190.9783Removal of antibiotic resistance genes and inactivation of antibiotic-resistant bacteria by oxidative treatments. The persistence of antibiotics in the environment because of human activities, such as seafood cultivation, has attracted great attention as they can give rise to antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB). In this study, we explored the inactivation and removal efficiencies of Escherichia coli SR1 and sul1 (plasmid-encoded ARGs), respectively, in their extracellular and intracellular forms (eARGs and iARGs) by three commonly used fishery oxidants, namely chlorine, bromine, and potassium permanganate (KMnO(4)), at the practical effective concentration range (0.5, 5, and 15 mg/L). Kinetics data were obtained using laboratory phosphate-buffered saline (PBS). Following the same fishery oxidation methods, the determined kinetics models were tested by studying the SR1 and sul1 disinfection efficiencies in (sterilized) pond water matrix. At concentrations of 5 and 15 mg/L, all three oxidants achieved sufficient cumulative integrated exposure (CT values) to completely inactivate SR1 and efficiently remove sul1 (up to 4.0-log). The oxidation methods were then applied to an unsterilized pond water matrix in order to study and evaluate the indigenous ARB and ARGs disinfection efficiencies in aquaculture, which reached 1.4-log and 1.0-log during treatment with fishery oxidants used in pond preparation at high concentrations before stocking (5-15 mg/L), respectively. A high chlorine concentration (15 mg/L) could efficiently remove ARGs (or iARGs) from pond water, and the iARG removal efficiency was higher than that of eARGs in pond water. The method and results of this study could aid in guiding future research and practical disinfection to control the spread of ARGs and ARB in aquaculture.202134030387