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786800.8604A 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
843110.8471A 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
120.8460Constructs for insertional mutagenesis, transcriptional signal localization and gene regulation studies in root nodule and other bacteria. Cassettes have been developed that contain an antibiotic resistance marker with and without a promoterless gusA reporter gene. The nptII (encoding kanamycin resistance) or aacCI (encoding gentamicin resistance) genes were equipped with the tac promoter (Ptac) and the trpA terminator (TtrpA) and then cloned between NotI sites to construct the CAS-Nm (Ptac-nptII-TtrpA) and CAS-Gm (Ptac/PaacCI-aacCI-TtrpA) cassettes. The markers were also cloned downstream to a modified promoterless Escherichia coli gusA gene (containing TGA stop codons in all three reading frames prior to its RBS and start codon) to construct the CAS-GNm (gusA-Ptac-nptII-TtrpA) or CAS-GGm (gusA-Ptac/PaacCI-aacCI-TtrpA) cassettes. Cassettes containing the promoterless gusA create type I fusions with a target DNA sequence to detect transcriptional activity. The promoterless gusA gene has also been cloned into a broad-host-range IncP1 plasmid. This construct will enable transcriptional activity to be monitored in different genetic backgrounds. Each cassette was cloned as a NotI fragment into the NotI site of a pUT derivative to construct four minitransposons. The mTn5-Nm (containing Ptac-nptII-TtrpA) and mTn5-Gm (containing Ptac/PaacCI-aacCI-TtrpA) minitransposons have been constructed specifically for insertional inactivation studies. The minitransposons mTn5-GNm (containing gusA-Ptac-nptII-TtrpA) and mTn5-GGm (containing gusA-Ptac/PaacCI-aacCI-TtrpA) can be used for transcription signal localization or insertional inactivation. The TAC-31R and TAC-105F primers can be used to sequence DNA flanking both sides of CAS-Nm, CAS-Gm, mTn5-Nm and mTn5-Gm. The WIL3 and TAC-105F primers can be used to sequence DNA flanking both sides of CAS-GNm, CAS-GGm, mTn5-GNm and mTn5-GGm. The specific application of these constructs to generate acid- or nodule-inducible fusions is presented. The new constructs provide useful tools for insertional mutagenesis, transcriptional signal localization and gene regulation studies in the root nodule bacteria and possibly other gram-negative bacteria.199910411257
782830.8432Simultaneous 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
840.8424The hawthorn CpLRR-RLK1 gene targeted by ACLSV-derived vsiRNA positively regulate resistance to bacteria disease. Virus-derived small interfering RNAs (vsiRNAs) can target not only viruses but also plant genes. Apple chlorotic leaf spot virus (ACLSV) is an RNA virus that infects Rosaceae plants extensively, including apple, pear and hawthorn. Here, we report an ACLSV-derived vsiRNA [vsiR1360(-)] that targets and down-regulates the leucine-rich repeat receptor-like kinase 1 (LRR-RLK1) gene of hawthorn (Crataegus pinnatifida). The targeting and cleavage of the CpLRR-RLK1 gene by vsiR1360(-) were validated by RNA ligase-mediated 5' rapid amplification of cDNA ends and tobacco transient transformation assays. And the CpLRR-RLK1 protein fused to green fluorescent protein localized to the cell membrane. Conserved domain and phylogenetic tree analyses showed that CpLRR-RLK1 is closely related to the proteins of the LRRII-RLK subfamily. The biological function of CpLRR-RLK1 was explored by heterologous overexpression of CpLRR-RLK1 gene in Arabidopsis. The results of inoculation of Pst DC3000 in Arabidopsis leaves showed that the symptoms of CpLRR-RLK1 overexpression plants infected with Pst DC3000 were significantly reduced compared with the wild type. In addition, the detection of reactive oxygen species and callose deposition and the expression analysis of defense-related genes showed that the CpLRR-RLK1 gene can indeed enhance the resistance of Arabidopsis to bacteria disease.202033180701
787050.8418Hierarchical Bi(2)O(2)CO(3) wrapped with modified graphene oxide for adsorption-enhanced photocatalytic inactivation of antibiotic resistant bacteria and resistance genes. There is growing pressure for wastewater treatment plants to mitigate the discharge of antibiotic resistant bacteria (ARB) and extracellular resistance genes (eARGs), which requires technological innovation. Here, hierarchical Bi(2)O(2)CO(3) microspheres were wrapped with nitrogen-doped, reduced graphene oxide (NRGO) for enhanced inactivation of multidrug-resistant E. coli NDM-1 and degradation of the plasmid-encoded ARG (bla(NDM-1)) in secondary effluent. The NRGO shell enhanced reactive oxygen species (ROS) generation (•OH and H(2)O(2)) by about three-fold, which was ascribed to broadened light absorption region (red-shifted up to 459 nm) and decreased electron-transfer time (from 55.3 to 19.8 ns). Wrapping enhanced E. coli adsorption near photocatalytic sites to minimize ROS scavenging by background constituents, which contributed to the NRGO-wrapped microspheres significantly outperforming commercial TiO(2) photocatalyst. ROS scavenger tests indicated that wrapping also changed the primary inactivation pathway, with photogenerated electron holes and surface-attached hydroxyl radicals becoming the predominant oxidizing species with wrapped microspheres, versus free ROS (e.g., •OH, H(2)O(2) and •O(2)(-)) for bare microspheres. Formation of resistance plasmid-composited microsphere complexes, primary due to the π-π stacking and hydrogen bonding between the shell and nucleotides, also minimized ROS scavenging and kept free plasmid concentrations below 10(2) copies/mL. As proof-of-concept, this work offers promising insight into the utilization of NRGO-wrapped microspheres for mitigating antibiotic resistance propagation in the environment.202032679343
784860.8417Simultaneous Removal of Antibiotic Resistant Bacteria, Antibiotic Resistance Genes, and Micropollutants by FeS(2)@GO-Based Heterogeneous Photo-Fenton Process. The co-occurrence of various chemical and biological contaminants of emerging concerns has hindered the application of water recycling. This study aims to develop a heterogeneous photo-Fenton treatment by fabricating nano pyrite (FeS(2)) on graphene oxide (FeS(2)@GO) to simultaneously remove antibiotic resistant bacteria (ARB), antibiotic resistance genes (ARGs), and micropollutants (MPs). A facile and solvothermal process was used to synthesize new pyrite-based composites. The GO coated layer forms a strong chemical bond with nano pyrite, which enables to prevent the oxidation and photocorrosion of pyrite and promote the transfer of charge carriers. Low reagent doses of FeS(2)@GO catalyst (0.25 mg/L) and H(2)O(2) (1.0 mM) were found to be efficient for removing 6-log of ARB and 7-log of extracellular ARG (e-ARG) after 30 and 7.5 min treatment, respectively, in synthetic wastewater. Bacterial regrowth was not observed even after a two-day incubation. Moreover, four recalcitrant MPs (sulfamethoxazole, carbamazepine, diclofenac, and mecoprop at an environmentally relevant concentration of 10 μg/L each) were completely removed after 10 min of treatment. The stable and recyclable composite generated more reactive species, including hydroxyl radicals (HO(•)), superoxide radicals (O(2)(• -)), singlet oxygen ((1)O(2)). These findings highlight that the synthesized FeS(2)@GO catalyst is a promising heterogeneous photo-Fenton catalyst for the removal of emerging contaminants.202235759741
785370.8414Natural pyrite and ascorbic acid co-enhance periodate activation for inactivation of antibiotic resistant bacteria and inhibition of resistance genes transmission: A green disinfection process dominated by singlet oxygen. The transmission of antibiotic resistance genes (ARGs) and the propagation of antibiotic resistant bacteria (ARB) threaten public health security and human health, and greener and more efficient disinfection technologies are expected to be discovered for wastewater treatment. In this study, natural pyrite and ascorbic acid (AA) were proposed as environmental-friendly activator and reductant for periodate (PI) activation to inactivate ARB. The disinfection treatment of PI/pyrite/AA system could inactivate 5.62 log ARB within 30 min, and the lower pH and higher PI and natural pyrite dosage could further boost the disinfection efficiency. The (1)O(2) and SO(4)(•-) were demonstrated to be crucial for the inactivation of ARB in PI/pyrite/AA system. The disinfection process destroyed the morphological structure of ARB, inducing oxidative stress and stimulating the antioxidant system. The PI/pyrite/AA system effectively reduced the intracellular and extracellular DNA concentration and ARGs abundance, inhibiting the propagation of ARGs. The presence of AA facilitated the activation of PI with natural pyrite and significantly increased the concentration of Fe(2+) in solution. The reusability of natural pyrite, the safety of the disinfection by-products and the inhibition of ARB regeneration indicated the application potential of PI/pyrite/AA system in wastewater disinfection.202439038380
1080.8410YODA Kinase Controls a Novel Immune Pathway of Tomato Conferring Enhanced Disease Resistance to the Bacterium Pseudomonas syringae. Mitogen-activated protein kinases (MAPK) play pivotal roles in transducing developmental cues and environmental signals into cellular responses through pathways initiated by MAPK kinase kinases (MAP3K). AtYODA is a MAP3K of Arabidopsis thaliana that controls stomatal development and non-canonical immune responses. Arabidopsis plants overexpressing a constitutively active YODA protein (AtCA-YDA) show broad-spectrum disease resistance and constitutive expression of defensive genes. We tested YDA function in crops immunity by heterologously overexpressing AtCA-YDA in Solanum lycopersicum. We found that these tomato AtCA-YDA plants do not show developmental phenotypes and fitness alterations, except a reduction in stomatal index, as reported in Arabidopsis AtCA-YDA plants. Notably, AtCA-YDA tomato plants show enhanced resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 and constitutive upregulation of defense-associated genes, corroborating the functionality of YDA in tomato immunity. This function was further supported by generating CRISPR/Cas9-edited tomato mutants impaired in the closest orthologs of AtYDA [Solyc08g081210 (SlYDA1) and Solyc03g025360 (SlYDA2)]. Slyda1 and Slyda2 mutants are highly susceptible to P. syringae pv. tomato DC3000 in comparison to wild-type plants but only Slyda2 shows altered stomatal index. These results indicate that tomato orthologs have specialized functions and support that YDA also regulates immune responses in tomato and may be a trait for breeding disease resistance.202033154763
786990.8407Nano-CeO(2) activates physical and chemical defenses of garlic (Allium sativum L.) for reducing antibiotic resistance genes in plant endosphere. The transmission of manure- and wastewater-borne antibiotic-resistant bacteria (ARB) to plants contributes to the proliferation of antimicrobial resistance in agriculture, necessitating effective strategies for preventing the spread of antibiotic resistance genes (ARGs) from ARB in the environment to humans. Nanomaterials are potential candidates for efficiently controlling the dissemination of ARGs. The present study investigated the abundance of ARGs in hydroponically grown garlic (Allium sativum L.) following nano-CeO(2) (nCeO(2)) application. Specifically, root exposure to nCeO(2) (1, 2.5, 5, 10 mg L(-1), 18 days) reduced ARG abundance in the endosphere of bulbs and leaves. The accumulation of ARGs (cat, tet, and aph(3')-Ia) in garlic bulbs decreased by 24.2-32.5 % after nCeO(2) exposure at 10 mg L(-1). Notably, the lignification extent of garlic stem-disc was enhanced by 10 mg L(-1) nCeO(2), thereby accelerating the formation of an apoplastic barrier to impede the upward transfer of ARG-harboring bacteria to garlic bulbs. Besides, nCeO(2) upregulated the gene expression related to alliin biosynthesis and increased allicin content by 15.9-16.2 %, promoting a potent antimicrobial defense for reducing ARG-harboring bacteria. The potential exposure risks associated with ARGs and Ce were evaluated according to the estimated daily intake (EDI). The EDI of ARGs exhibited a decrease exceeding 95 %, while the EDI of Ce remained below the estimated oral reference dose. Consequently, through stimulating physical and chemical defenses, nCeO(2) contributed to a reduced EDI of ARGs and Ce, highlighting its potential for controlling ARGs in plant endosphere within the framework of nano-enabled agrotechnology.202438570269
8492100.8407Promotion effects and mechanisms of molybdenum disulfide on the propagation of antibiotic resistance genes in soil. The rapid development of nanotechnology has aroused considerable attentions toward understanding the effects of engineered nanomaterials (ENMs) on the propagation of antibiotic resistance. Molybdenum disulfide (MoS(2)) is an extensively used ENM and poses potential risks associated with environmental exposure; nevertheless, the role of MoS(2) toward antibiotic resistance genes (ARGs) transfer remains largely unknown. Herein, it was discovered that MoS(2) nanosheets accelerated the horizontal transfer of RP4 plasmid across Escherichia coli in a dose-dependent manner (0.5-10 mg/L), with the maximum transfer frequency 2.07-fold higher than that of the control. Integration of physiological, transcriptomics, and metabolomics analyses demonstrated that SOS response in bacteria was activated by MoS(2) due to the elevation of oxidative damage, accompanied by cell membrane permeabilization. MoS(2) promoted bacterial adhesion and intercellular contact via stimulating the secretion of extracellular polysaccharides. The ATP levels were maximally increased by 305.7 % upon exposure to MoS(2), and the expression of plasmid transfer genes was up-regulated, contributing to the accelerated plasmid conjugation and increased ARG abundance in soil. Our findings highlight the roles of emerging ENMs (e.g., MoS(2)) in ARGs dissemination, which is significant for the safe applications and risk management of ENMs under the development scenarios of nanotechnology.202337062264
7829110.8403Insights 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
7989120.8401Feasibility of sulfate-calcined eggshells for removing pathogenic bacteria and antibiotic resistance genes from landfill leachates. High abundance of human pathogen and antibiotic resistance genes (ARGs) in landfill leachate has become an emerging threat against human health. Therefore, sulfate- and calcination-modified eggshells as green agricultural bioresource were applied to test the feasibility of removing pathogenic bacteria and ARGs from leachate. The highest removal of Escherichia coli (E. coil) and gentamycin resistant gene (gmrA) from artificial contaminated landfill leachate was achieved by the application of eggshell with combined treatment of sulfate and calcination. The 16S and gmrA gene copies of E. coil declined significantly from 1.78E8±8.7E6 and 4.12E8±5.9E6 copies mL(-1) to 1.32E7±2.6E6 and 2.69E7±7.2E6 copies mL(-1), respectively, within 24h dynamic adsorption equilibrium process (p<0.05). Moreover, according to the Langmuir kinetic model, the greatest adsorption amount (1.56×10(9) CFU E. coil per gram of modified eggshells) could be obtained at neutral pH of 7.5. The optimal adsorption eggshells were then screened to the further application in three typical landfill leachates in Nanjing, eastern China. Significant decrease in species and abundance of pathogenic bacteria and ARGs (tet, sul, erm, qnr, and ampC) indicated its great efficiency to purify landfill leachates. This study demonstrated that sulfate-calcined eggshells can be an environmentally-friendly and highly efficient bioadsorbent to the management of reducing dissemination risk of pathogen and ARGs in landfill leachate.201728343745
523130.8397Sulfide-carbonate-mineralized functional bacterial consortium for cadmium removal in flue gas. Sulfide-carbonate-mineralized functional bacterial consortium was constructed for flue gas cadmium biomineralization. A membrane biofilm reactor (MBfR) using the bacterial consortium containing sulfate reducing bacteria (SRB) and denitrifying bacteria (DNB) was investigated for flue gas cadmium (Cd) removal. Cadmium removal efficiency achieved 90%. The bacterial consortium containing Citrobacter, Desulfocurvus and Stappia were dominated for cadmium resistance-nitrate-sulfate reduction. Under flue gas cadmium stress, ten cadmium resistance genes (czcA, czcB, czcC, czcD, cadA, cadB, cadC, cueR, copZ, zntA), and seven genes related to sulfate reduction, increased in abundance; whereas others, nine genes related to denitrification, decreased, indicating that cadmium stress was advantageous to sulfate reduction in the competition with denitrification. A bacterial consortium could capable of simultaneously cadmium resistance, sulfate reduction and denitrification. Microbial induced carbonate precipitation (MICP) and biological adsorption process would gradually yield to sulfide-mineralized process. Flue gas cadmium could transform to Cd-EPS, cadmium carbonate (CdCO(3)) and cadmium sulfide (CdS) bioprecipitate. The functional bacterial consortium was an efficient and eco-friendly bifunctional bacterial consortium for sulfide-carbonate-mineralized of cadmium. This provides a green and low-carbon advanced treatment technology using sulfide-carbonate-mineralized functional bacterial consortium for the removal of cadmium or other hazardous heavy metal contaminants in flue gas.202439019186
7861140.8394The removal of antibiotic resistant bacteria and genes and inhibition of the horizontal gene transfer by contrastive research on sulfidated nanoscale zerovalent iron activating peroxymonosulfate or peroxydisulfate. Antibiotic resistant bacteria (ARB) and the antibiotic resistance genes (ARGs) dissemination via plasmid-mediated conjugation have attracted considerable attentions. In this research, sulfidated nanoscale zerovalent iron (S-nZVI)/peroxymonosulfate (PMS) and S-nZVI/peroxydisulfate (PDS) process were investigated to inactivate ARB (Escherichia coli DH5α with RP4 plasmid, Pseudomonas. HLS-6 contains sul1 and intI1 on genome DNA sequence). S-nZVI/PMS system showed higher efficiency than S-nZVI/PDS on ARB inactivation. Thus, the optimal condition 28 mg/L S-nZVI coupled with 153.7 mg/L (0.5 mM) PMS was applied to remove both intracellular ARGs (iARGs) and ARB. The oxidative damage of ARB cell was systemically studied by cell viability, intracellular Mg(2+) levels, the changes of extracellular and internal structure, integrity of cell walls and membranes and enzymatic activities. S-nZVI/PMS effectively inactivated ARB (~7.32 log) within 15 min. These effects were greatly higher than those achieved individually. Moreover, removal efficiencies of iARGs sul1, intI1 and tetA were 1.52, 1.79 and 1.56 log, respectively. These results revealed that S-nZVI and PMS have a synergistic effect against ARB and iARGs. The regrowth assays illustrated that the ARB were effectively inactivated. By verifying the inhibitory impacts of S-nZVI/PMS treatment on conjugation transfer, this work highlights a promising alternative technique for inhibiting the horizontal gene transfer.202234482079
7859150.8394Abatement of antibiotics and resistance genes during catalytic ozonation enhanced sludge dewatering process: Synchronized in volume and hazardousness reduction. Based on the efficiency of the catalytic ozonation techniques (HDWS+O(3) and MnFe(2)O(4) @SBC+O(3)) in enhancing the sludge dewaterability, the effectiveness in synchronized abatement antibiotics and antibiotic resistance genes (ARGs) was conducted to determine. The results revealed that catalytic ozonation conditioning altered the distribution of target antibiotics (tetracycline (TC), oxytetracycline (OTC), norfloxacin (NOR), ofloxacin (OFL)) in the dewatered filtrate, the dewatered sludge cake and the extra-microcolony/cellular polymers (EMPS/ECPS) layers, achieving the redistribution from solid-phase adsorption to liquid-phase dissolution. The total degradation rate was over 90% for TC and OTC, 72-78% for NOR and OFL; the abatement efficiency of eleven ARGs reached 1.47-3.01 log and 1.64-3.59 log, respectively, and more than four eARGs were eliminated. The effective abatement of the absolute abundance of Mobile genetic elements (MGEs) (0.91-1.89 log) demonstrated that catalytic ozonation conditioning could also significantly inhibit horizontal gene transfer (HGT). The abundance of resistant bacteria was greatly reduced and the signal transduction of the typical ARGs host bacteria was inhibited. The highly reactive oxidation species (ROS) generated were responsible for the abatement of antibiotics and ARGs. These findings provided new insights into the sludge conditioning for ideal and synchronized reduction in volume and hazardousness by catalytic ozonation processes in sludge treatment.202437944236
7855160.8393Combat against antibiotic resistance genes during photo-treatment of magnetic Zr-MOFs@Layered double hydroxide heterojunction: Conjugative transfer risk mitigating and bacterial inactivation. The dissemination of antimicrobial resistance (AMR) in wastewater treatment poses a severe threat to the global ecological environment. This study explored the effectiveness of photocatalysis in inactivating antibiotic resistant bacteria (ARB) and quantitatively clarified the inhibiting rate of the transfer of antibiotics resistance genes (ARGs). Herein, the magnetic heterojunction as UiO-66-NH(2)@CuFe LDH-Fe(3)O(4) (UN-66@LDH-Fe) effectively facilitated the electron-hole separation and accelerated the photogenerated charge transfer, thereby guaranteeing the stable practical application in aeration tanks. Notably, the internal electric field of heterogeneous photocatalyst resulted in significant increase of ARGs inactivation, achieving 5.63 log of ARB, 3.66 log of tetA and 3.57 log of Ampr genes were photodegraded under optimal reaction conditions within 6 h. Based on the complex microbial and molecular mechanism of multiple-ARB communities inactivation in photo-treatment, the photogenerated reactive oxygen species (ROSs, ·OH and ·O(2)(-)) effectively destroyed bacterial membrane protein, thereby the intracellular ROSs and redox cycles further induced oxidative stress, attributing to the abundance reduction of ARGs and their host bacteria. Moreover, long-term (7 days) continuous operation preliminarily verified the practical potential in reducing AMR spread and developing wastewater treatment efficacy. Overall, this study presented an advantageous synergistic strategy for mitigating the AMR-associated environmental risk in wastewater treatment.202540188541
392170.8391Stable Tagging of Rhizobium meliloti with the Firefly Luciferase Gene for Environmental Monitoring. A system for stable tagging of gram-negative bacteria with the firefly luciferase gene, luc, is described. A previously constructed fusion constitutively expressing luc from the lambdap(R) promoter was used. Stable integration into the bacterial genome was achieved by use of mini-Tn5 delivery vectors. The procedure developed was applied for tagging of representative gram-negative bacteria, such as Escherichia coli, Rhizobium meliloti, Pseudomonas putida, and Agrobacterium tumefaciens. The system permitted the detection of tagged R. meliloti in the presence of more than 10 CFU per plate without the use of any selective markers (such as antibiotic resistance genes). No significant differences in growth rates or soil survival were found between the marked strain and the wild-type strain. Studies of bioluminescent R. meliloti also revealed a good correlation between cell biomass and bioluminescence. The firefly luciferase tagging system is an easy, safe, and sensitive method for the detection and enumeration of bacteria in the environment.199316349015
7849180.8391Efficient removal of antibiotic-resistant bacteria and intracellular antibiotic resistance genes by heterogeneous activation of peroxymonosulfate on hierarchical macro-mesoporous Co(3)O(4)-SiO(2) with enhanced photogenerated charges. Antibiotic resistance genes (ARGs) and their host antibiotic-resistant bacteria (ARB) are widely detected in the environment and pose a threat to human health. Traditional disinfection in water treatment plants cannot effectively remove ARGs and ARB. This study explored the potential of a heterogeneous photo-Fenton-like process utilizing a hierarchical macro-mesoporous Co(3)O(4)-SiO(2) (MM CS) catalyst for activation of peroxymonosulfate (PMS) to inactivate ARB and degrade the intracellular ARGs. A typical gram-negative antibiotic-resistant bacteria called Pseudomonas sp. HLS-6 was used as a model ARB. A completed inactivation of ARB at ∼10(7) CFU/mL was achieved in 30 s, and an efficient removal rate of more than 4.0 log for specific ARGs (sul1 and intI1) was achieved within 60 min by the MM CS-based heterogeneous photo-Fenton-like process under visible light and neutral pH conditions. Mechanism investigation revealed that •O(2)(-) and (1)O(2) were the vital reactive species for ARB inactivation and ARG degradation. The formation and transformation of the active species were proposed. Furthermore, the hierarchical macro-mesoporous structure of MM CS provided excellent optical and photoelectrochemical properties that promoted the cycle of Co(3+)/Co(2+) and the effective utilization of PMS. This process was validated to be effective in various water matrices, including deionized water, underground water, source water, and secondary effluent wastewater. Collectively, this work demonstrated that the MM CS-based heterogeneous photo-Fenton-like process is a promising technology for controlling the spread of antibiotic resistance in aquatic environments.202235149504
7831190.8391Integration of nanowire-confined electroporation of antibiotic-resistant bacteria and electroactivation of peracetic acid for eliminating intracellular resistance genes. Antimicrobial resistance is one of the most substantial challenges for global public health. To address the inefficient elimination of intracellular resistance genes (i-ARGs) in antibiotic-resistant bacteria (ARB) by peracetic acid (PAA) oxidation, we developed an integration strategy (NW-EP/EA) of nanowire-confined electroporation (NW-EP) of ARB cells and nanowire-confined electroactivation (NW-EA) of PAA with a sequential oxidation-reduction process. The locally enhanced electric field and electrocatalytic activity over NW tips prompted the formation of electroporation pores on ARB cells and the generation of reactive ⋅OH and RO⋅ radicals by PAA electroactivation. The NW-EP/EA with Pd-coated TiO(2)NW cathode with atomic H* evolution exhibited 0.6 -2.8-log higher i-ARG removal than the pristine TiO(2)NW cathode, especially achieving ∼5.0-log i-ARG removal (99.999 %) at 4.0 V and 2.0 mM PAA with ∼4.1-log synergistic effect and ∼10 times lower energy consumption as compared with the individual NW-EP (∼0.32-log and 52.1 %) and PAA (∼0.56-log and 74.4 %). For the sequential oxidation-reduction process, the electrooxidative activation of PAA on TiO(2)NW anode produced H(+) ions, ⋅OH and RO⋅ radicals for enlarging electroporation pores, and the generated H(+) ions promoted the evolution of atomic H* and electroreduction of PAA on subsequent Pd-TiO(2)NW cathode for further facilitating ARB cell damages, i-ARG leakage and degradation. The effective i-ARGs removal and HGT inhibition in tap water suggested the great application potentials of NW-EP/EA in the control of ARGs dissemination risks in drinking water.202540907311