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779600.9155Irreversible inactivation of carbapenem-resistant Klebsiella pneumoniae and its genes in water by photo-electro-oxidation and photo-electro-Fenton - Processes action modes. Carbapenem-resistant Klebsiella pneumoniae is a critical priority pathogen according to the World Health Organization's classification. Effluents of municipal wastewater treatment plants (EWWTP) may be a route for K. pneumoniae dissemination. Herein, the inactivation of this microorganism in simulated EWWTP by the photo-electro-oxidation (PEO) and photo-electro-Fenton (PEF) processes was evaluated. Firstly, the disinfecting ability and action pathways of these processes were established. PEO achieved faster K. pneumoniae inactivation (6 log units in 75 min of treatment) than the PEF process (6 log units in 105 min of treatment). PEO completely inactivated K. pneumoniae due to the simultaneous action of UVA light, electrogenerated H(2)O(2,) and anodic oxidation pathways. The slower inactivation of K. pneumoniae when using PEF was related to interfering screen effects of iron oxides on light penetration and the diffusion of the bacteria to the anode. However, both PEO and PEF avoided the recovery and regrowth of treated bacteria (with no detectable increase in the bacteria concentration after 24 h of incubation). In addition to the bacteria evolution, the effect of treatment processes on the resistance gene was examined. Despite inactivation of K. pneumoniae by PEF was slower than by PEO, the former process induced a stronger degrading action on the gene, conferring the resistance to carbapenems (PEF had a Ct value of 24.92 cycles after 105 min of treatment, while PEO presented a Ct of 19.97 cycles after 75 min). The results of this research indicate that electrochemical processes such as PEO and PEF are highly effective at dealing with resistant K. pneumoniae in the EWWTP matrix.202134146813
10110.9114The encapsulated strain TIGR4 of Streptococcus pneumoniae is phagocytosed but is resistant to intracellular killing by mouse microglia. The polysaccharide capsule is a major virulence factor of Streptococcus pneumoniae as it confers resistance to phagocytosis. The encapsulated serotype 4 TIGR4 strain was shown to be efficiently phagocytosed by the mouse microglial cell line BV2, whereas the type 3 HB565 strain resisted phagocytosis. Comparing survival after uptake of TIGR4 or its unencapsulated derivative FP23 in gentamicin protection and phagolysosome maturation assays, it was shown that TIGR4 was protected from intracellular killing. Pneumococcal capsular genes were up-regulated in intracellular TIGR4 bacteria recovered from microglial cells. Actual presence of bacteria inside BV2 cells was confirmed by transmission electron microscopy (TEM) for both TIGR4 and FP23 strains, but typical phagosomes/phagolysosomes were detected only in cells infected with the unencapsulated strain. In a mouse model of meningitis based on intracranic inoculation of pneumococci, TIGR4 caused lethal meningitis with an LD(50) of 2 × 10² CFU, whereas the LD(50) for the unencapsulated FP23 was greater than 10⁷ CFU. Phagocytosis of TIGR4 by microglia was also demonstrated by TEM and immunohistochemistry on brain samples from infected mice. The results indicate that encapsulation does not protect the TIGR4 strain from phagocytosis by microglia, while it affords resistance to intracellular killing.201020615478
820.9070The 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
786030.9069Enhanced 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
783540.9068Crouching bacteria, hidden tetA genes in natural waters: Intracellular damage via double persulfate activation (UVA/Fe(2+)/PDS) effectively alleviates the spread of antibiotic resistance. In this study, we elucidated the chemical and biological inactivation mechanisms of peroxydisulfate (PDS) activated by UVA and Fe(2+) (UVA/Fe(2+)/PDS) in wild-type antibiotic-resistant bacteria (ARB) isolated from a river in Inner Mongolia. Among the screened wild-type ARB, the relative abundance of unidentified Enterobacteriaceae, Stenotrophomonas, and Ralstonia was high. A ratio of 1:1 for Fe(2+) and PDS under 18 W·m(-2) UVA radiation (sunny days) completely inactivated the environmental ARB isolates. In the macro view of the inactivation process, Fe(2+) first activates PDS rapidly, and later the UVA energy accumulated starts to activate PDS; HO• then becomes the main active species at a rate-limiting step. From a micro perspective, damage to the cell wall, intracellular proteins, inactivation of antioxidant enzymes, and genetic material degradation are the inactivation series of events by UVA/Fe(2+)/PDS, contributing to the 97.8 % inactivation of ARB at the initial stage. No regrowth of sublethal ARBs was observed. The transfer of tetracycline resistance genes from ARB to lab E. coli was evaluated by horizontal gene transfer (HGT), in which no HGT occurred when ARB was eliminated by UVA/Fe(2+)/PDS. Moreover, the sulfate and iron residuals in the effluents of treated water were lower than the drinking water standards. In summary, PDS, UVA, and Fe(2+) activation effectively inactivated wild ARB with a low concentration of reagents, while inhibiting their regrowth and spread of resistance due to the contribution of intracellular inactivation pathways.202439316921
787650.9065Sulfamethoxazole impact on pollutant removal and microbial community of aerobic granular sludge with filamentous bacteria. In this study, sulfamethoxazole (SMX) was employed to investigate its impact on the process of aerobic granule sludge with filamentous bacteria (FAGS). FAGS has shown great tolerance ability. FAGS in a continuous flow reactor (CFR) could keep stable with 2 μg/L of SMX addition during long-term operation. The NH(4)(+), chemical oxygen demand (COD), and SMX removal efficiencies kept higher than 80%, 85%, and 80%, respectively. Both adsorption and biodegradation play important roles in SMX removal for FAGS. The extracellular polymeric substances (EPS) might play important role in SMX removal and FAGS tolerance to SMX. The EPS content increased from 157.84 mg/g VSS to 328.22 mg/g VSS with SMX addition. SMX has slightly affected on microorganism community. A high abundance of Rhodobacter, Gemmobacter, and Sphaerotilus of FAGS may positively correlate to SMX. The SMX addition has led to the increase in the abundance of the four sulfonamide resistance genes in FAGS.202336871701
786160.9065The 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
784870.9062Simultaneous 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
788580.9061Susceptibility, resistance and resilience of anammox biomass to nanoscale copper stress. The increasing use of engineered nanoparticles (NPs) poses an emerging challenge to biological wastewater treatment. The long-term impact of CuNPs on anaerobic ammonium oxidation (anammox) process was firstly investigated in this study. The nitrogen removal capacity of anammox reactor was nearly deprived within 30days under the stress of 5.0mgL(-1) CuNPs and the relative abundance of anammox bacteria (Ca. Kuenenia) was decreased from 29.59% to 17.53%. Meanwhile, copper resistance genes associated with the Cus, Cop and Pco systems were enriched to eliminate excess intracellular copper. After the withdrawal of CuNPs from the influent, the nitrogen removal capacity of anammox biomass recovered completely within 70days. Overall, anammox biomass showed susceptibility, resistance and resilience to the stress of CuNPs. Therefore, the potential impacts of ENPs on anammox-based processes should be of great concern.201728550773
33490.9058Transformation of soybean protoplasts from permanent suspension cultures by cocultivation with cells of Agrobacterium tumefaciens. Cell wall regenerating protoplasts from soybean cells kept in suspension culture were cocultivated with bacteria which were derived from the nopaline strain C58 of Agrobacterium tumefaciens. When the bacteria carried an oncogenic Ti-plasmid, about 5% of the surviving protoplasts were able to form calli on hormone-free agar in contrast to controls, where bacteria without Ti-plasmid were applied, and where no calli were formed. After isolation of DNA from hormone-independently growing cells further evidence for transformation was obtained by hybridization to Ti-plasmid specific RNA and by rescue of a segment with a bacterial resistance gene which had been inserted before into the T-DNA. Transfer of T-DNA harboring a neomycin-resistance gene activated by the nos-promoter resulted in calli growing on kanamycin. Verification of segments located at the left and the right part of the T-DNA indicated the presence of its entire length in transformed soybean cells. Expression of T-DNA genes was measured by the assay of nopaline-synthase. Cells cultured on agar had a much higher level of nopaline-synthase than fast growing cells in suspension culture. Transferring them to agar or treatment with azacytidine strongly increased synthesis of nopaline-synthase indicating a reversible repression presumably via a methylation mechanism.198724276903
7828100.9057Simultaneous 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
528110.9057Effect of dimethyl sulphoxide on the expression of nitrogen fixation in bacteria. Storage in dimethyl sulphoxide (DMSO) of Escherichia coli K12 hybrids carrying nif+ genes from Klebsiella pneumoniae can result in selection of a defective nitrogen-fixing phenotype. Similar results are obtained with E. coli K12 hybrids containing the nitrogen-fixing capacity from Rhizobium trifolii. DMSO appears to affect particular inner membrane proteins associated with energy metabolism in E. coli K12 and four chromosomal regions (chlD, chlG, his and unc) are associated with resistance to DMSO.1977332135
530120.9057Location of the genes for anthranilate synthase in Streptomyces venezuelae ISP5230: genetic mapping after integration of the cloned genes. The anthranilate synthase (trpEG) genes in Streptomyces venezuelae ISP5230 were located by allowing a segregationally unstable plasmid carrying cloned S. venezuelae trpEG DNA and a thiostrepton resistance (tsr) marker to integrate into the chromosome. The integrated tsr was mapped by conjugation and transduction to a location close to tyr-2, between arg-6 and trpA13. A genomic DNA fragment containing trpC from S. venezuelae ISP5230 was cloned by complementation of a trpC mutation in Streptomyces lividans. Evidence from restriction enzyme analysis of the cloned DNA fragments, from Southern hybridization using the cloned trp DNA as probes, and from cotransduction frequencies, placed trpEG at a distance of 12-45 kb from the trpCBA cluster. The overall arrangement of tryptophan biosynthesis genes in the S. venezuelae chromosome differs from that in other bacteria examined so far.19938515229
7827130.9055Inactivation of antibiotic-resistant bacteria and antibiotic resistance genes by electrochemical oxidation/electro-Fenton process. Antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the environment are of great concern due to their potential risk to human health. The effluents from wastewater treatment plants and livestock production are major sources of ARB and ARGs. Chlorination, UV irradiation, and ozone disinfection cannot remove ARGs completely. In this study, the potential of electrochemical oxidation and electro-Fenton processes as alternative treatment technologies for inactivation of ARB and ARGs in both intracellular and extracellular forms was evaluated. Results showed that the electrochemical oxidation process was effective for the inactivation of selected ARB but not for the removal of intracellular ARGs or extracellular ARGs. The electro-Fenton process was more effective for the removal of both intracellular and extracellular ARGs. The removal efficiency after 120 min of electro-Fenton treatment under 21.42 mA/cm(2) was 3.8 logs for intracellular tetA, 4.1 logs for intracellular ampC, 5.2 logs for extracellular tetA, and 4.8 logs for extracellular ampC, respectively in the presence of 1.0 mmol/L Fe(2+). It is suggested that electrochemical oxidation is an effective disinfection method for ARB and the electro-Fenton process is a promising technology for the removal of both intracellular and extracellular ARGs in wastewater.202032701499
540140.9055Effect of ogt expression on mutation induction by methyl-, ethyl- and propylmethanesulphonate in Escherichia coli K12 strains. We have previously reported the isolation of an Escherichia coli K12 mutant that is extremely sensitive to mutagenesis by low doses of ethylating agents. We now show by Southern analysis that the mutation involves a gross deletion covering at least the ogt and fnr genes and that no O6-alkylguanine-DNA-alkyltransferase activity is present in cell-free extracts of an ada::Tn10 derivative of these bacteria. Confirmation that sensitisation to ethylation-induced mutagenesis was attributable to ogt and not to any other loci covered by the deletion was obtained by constructing derivatives. Thus an ogt::kanr disruption mutation was introduced into the parental ogt+ bacteria, and the ogt::kanr mutation was then eliminated by cotransduction of ogt+ with the closely linked Tetr marker (zcj::Tn10). The delta(ogt-fnr) deletion or ogt::kanr disruption mutants were highly sensitive to ethyl methanesulphonate-induced mutagenesis, as measured by the induction of forward mutations to L-arabinose resistance (Arar). Furthermore, the number of Arar mutants increased linearly with dose, unlike the case in ogt+ bacteria, which had a threshold dose below which no mutants accumulated. Differences in mutability were even greater with propyl methanesulphonate. Overproduction of the ogt alkyltransferase from a multicopy plasmid reduced ethylmethanesulphonate-induced mutagenesis in the ogt- mutant strains and also methylmethanesulphonate mutagenesis in ada- bacteria. A sample of AB1157 obtained from the E. coli K12 genetic stock centre also had a deletion covering the ogt and fnr genes. Since such deletions greatly influence the mutagenic responses to alkylating agents, a survey of the presence of the ogt gene in the E. coli K12 strain being used is advisable.19948152424
9061150.9052Ozone-Resistant Bacteria, an Inconvenient Hazard in Water Reclamation: Resistance Mechanism, Propagating Capacity, and Potential Risks. Resistant bacteria have always been of research interest worldwide. In the urban water system, the increased disinfectant usage gives more chances for undesirable disinfection-resistant bacteria. As the strongest oxidative disinfectant in large-scale water treatment, ozone might select ozone-resistant bacteria (ORB), which, however, have rarely been reported and are inexplicit for their resistant mechanisms and physiological characteristics. In this study, six strains of ORB were screened from a water reclamation plant in Beijing. Three of them (O7, CR19, and O4) were more resistant to ozone than all previously reported ORB or even spores. The ozone consumption capacity of extracellular polymeric substances and cell walls was proved to be the main sources of bacterial ozone resistance, rather than intracellular antioxidant enzymes. The transcriptome results elucidated that strong ORB possessed a combined antioxidant mechanism consisting of the enhanced transcription of protein synthesis, protein export, and polysaccharide export genes (LptF, LptB, NodJ, LivK, LviG, MetQ, MetN, and GltU). This study confirmed the existence of ORB in urban water systems and brought doubts to the idea of a traditional control strategy against chlorine-resistant bacteria. A salient "trade-off" effect between the ozone resistance and propagation ability indicated the weakness and potential control approaches of ORB.202439344972
7881160.9052Bacterial community shift and antibiotics resistant genes analysis in response to biodegradation of oxytetracycline in dual graphene modified bioelectrode microbial fuel cell. This study explored the biodegradation mechanisms of oxytetracycline (OTC/O) and electrochemical characteristics from the perspective of bacterial community shift and OTC resistance genes in dual graphene modified bioelectrode microbial fuel cell (O-D-GM-BE MFC). In phylum level, Proteobacteria was accounted to 95.04% in O-GM-BA, Proteobacteria and Bacteroidetes were accounted to 59.13% and 20.52% in O-GM-BC, which were beneficial for extracellular electron transport (EET) process and OTC biodegradation. In genus level, the most dominant bacteria in O-GM-BA were Salmonella and Trabulsiella, accounting up to 83.04%, moreover, representative exoelectrogens (Geobacter) were enriched, which contributed to OTC biodegradation and electrochemical performances; abundant degrading bacteria (Moheibacter, Comamonas, Pseudomonas, Dechloromonas, Nitrospira, Methylomicrobium, Pseudorhodoferax, Thiobacillus, Mycobacterium) were enriched in O-GM-BC, which contributed to the maximum removal efficiency of OTC; coding resistance genes of efflux pump, ribosome protective protein and modifying or passivating were all found in O-GM-BE, and this explained the OTC removal mechanisms from gene level.201930640017
6150170.9052Redox biotransformation of arsenic along with plant growth promotion by multi-metal resistance Pseudomonas sp. MX6. Remediation of toxic metal-polluted sites by microorganisms is an environment-friendly remediation technique. Multi-metal-resistant bacteria were isolated from a wastewater treatment plant showing resistance against As(III), As(V), Cr, Co, Cu, Cd, Hg, Ni, Pb, Se and Zn. Maximum resistance against all metals was shown by the bacterial isolate MX-6 (As 20mM, Cd 30mM, Cr 5.0mM, Co 25mM, Cu 25mM, Ni 20mM, Zn 30mM, Pb 15mM, Se 20mM and Hg 2.5mM), which was identified as Pseudomonas sp. through 16S rDNA sequencing. Pseudomonas sp. MX-6 reduced 506μM As(V) and also oxidized 160μM As(III). The genes for As, Cd, Se and Zn resistance in Pseudomonas sp. MX-6 were found to be plasmid borne, as indicated by transformation. Pseudomonas sp. MX-6 produced 49.37μg·mL(-1) IAA and was also positive for HCN production and phosphate solubilisation. The bacterial isolate also supported Vigna radiata growth, both in the absence and presence of the aforementioned metals. Such bacteria can be used as biofertilizers to reclaim the polluted lands and to enhance crop production in metal-contaminated soils.201728684222
7836180.9051Efficient Degradation of Intracellular Antibiotic Resistance Genes by Photosensitized Erythrosine-Produced (1)O(2). Intracellular antibiotic resistance genes (iARGs) constitute the important part of wastewater ARGs and need to be efficiently removed. However, due to the dual protection of intracellular DNA by bacterial membranes and the cytoplasm, present disinfection technologies are largely inefficient in iARG degradation. Herein, we for the first time found that erythrosine (ERY, an edible dye) could efficiently degrade iARGs by producing abundant (1)O(2) under visible light. Seven log antibiotic-resistant bacteria were inactivated within only 1.5 min, and 6 log iARGs were completely degraded within 40 min by photosensitized ERY (5.0 mg/L). A linear relationship was established between ARG degradation rate constants and (1)O(2) concentrations in the ERY photosensitizing system. Surprisingly, a 3.2-fold faster degradation of iARGs than extracellular ARGs was observed, which was attributed to the unique indirect oxidation of iARGs induced by (1)O(2). Furthermore, ERY photosensitizing was effective for iARG degradation in real wastewater and other photosensitizers (including Rose Bengal and Phloxine B) of high (1)O(2) yields could also achieve efficient iARG degradation. The findings increase our knowledge of the iARG degradation preference by (1)O(2) and provide a new strategy of developing technologies with high (1)O(2) yield, like ERY photosensitizing, for efficient iARG removal.202337531556
524190.9050Sulfamethoxazole degradation by Pseudomonas silesiensis F6a isolated from bioelectrochemical technology-integrated constructed wetlands. The antibiotic-degrading ability and mechanism of the bacteria in the novel and ecological bioelectrochemical technology-integrated constructed wetlands (BICW) remain unknown. In this study, the sulfamethoxazole (SMX) degrading strain Pseudomonas silesiensis F6a (F6a), which had high degradation efficiency, was firstly isolated from a substrate sample in BICW. The SMX degradation process of F6a follows pseudo first order kinetics. Four metabolic pathways and twelve degradation products were identified. Based on genomics and proteomics analysis, six key SMX-degrading genes, Gene4641 deoC, Gene0552 narI, Gene0546 luxS, Gene1753 nuoH, Gene0655 and Gene4650, were identified, which were mainly participated in C-S cleavage, S-N hydrolysis and isoxazole ring cleavage. Interestingly, we found the corresponding sulfonamides resistance genes were not detected in F6a, which may provide an evidence for low abundance of the sulfonamides resistance genes in BICW system. These findings would contribute to a better understanding of biotransformation of antibiotic in the BICW.202235636241