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ObjectiveAntibiotic resistance genes （ARGs） have received wide attention all over the world. The purpose of this study was to explore the bacterial community structure， the types and levels of antibiotic resistance genes in a water body in east China， and to compare and analyze the characteristics of microbial species distribution and antibiotic resistance gene distribution in various water environments. MethodsA total of 10 households in Haimen City， Jiangsu Province were selected and their surrounding water environment samples were collected. 21 water samples including river water （4）， Mingou water （9） and well water （8） were collected for metagenomics sequencing， assembled with MetaWRAP， annotated with CARD database， and analyzed with R software. ResultsIn various water bodies， the dominant bacteria phyla was Proteobacteria， the dominant bacteria genera were Deuterostomia， Pseudomonas， Flavobacteriales and Streptomycetaceae. The ARGs annotated were mainly composed of quinolones， aminoglycosides， macrolides and beta-lactams antibiotic resistance genes. The top four relative abundance of resistance genes were macB， RanA， evgS and TxR， The average absolute abundance and expression of resistance genes in well water and Mingou water were higher than those in river water. ConclusionMultiple ARGs are detected to varying degrees in well water， river water， and Mingou water bodies， and the expression of resistance genes in well water and Mingou water bodies is higher than that in river water bodies， possibly due to human production and living activities.
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ObjectiveTo determine the distribution of various antibiotic resistance genes （ARGs） in raw water from drinking water source， and to explore the correlation between the ARGs and common carbapenem-resistant and multidrug-resistant bacteria isolated from drinking water source， so as to provide scientific evidence for improving the safety of urban drinking water. MethodsA total of 30 raw water samples were collected from a major drinking water source in Shanghai in 2020. Bacterial strains were selectively cultured on Columbia blood agar medium containing 1 μg·μL-1 meropenem， and then identified by MALDI-TOF-MS mass spectrometry system. Minimum inhibitory concentration （MIC） of the strains was detected by broth microdilution method. The water samples were filtered through a 0.45 μm filter membrane and diversity of ARGs was determined by using high-throughput metagenomic sequencing. ResultsA total of 64 strains of carbapenem-resistant bacteria were isolated from the water samples， including Stenotrophomonas maltophilia， Enterococcus faecium， Pseudomonas aeruginosa， Acinetobacter baumannii and Klebsiella pneumoniae， which were resistant to a variety of common antibiotics. Using metagenomic sequencing，1 244 ARGs were identified. The relative average abundance of the top 100 ARGs accounted for 96.1%， and that of the multidrug-resistant ARGs accounted for 63.41%. Furthermore， the multidrug-resistant ARGs were mainly adeJ， mexT， adeC， oprM， mexF， mdfA， mexB， mdtK， adeK， etc. Using Spearman's correlation， five multidrug-resistant bacteria isolated from the drinking water source were significantly associated with the ARGs. ConclusionRelative abundance of multidrug-resistant ARGs is high in raw water from main drinking water source. The five isolated carbapenem-resistant and multidrug-resistant bacteria are significantly correlated with the ARGs. It warrants strengthening the rational and standardized application of antibiotics to protect water resources and ensure the safety of drinking water.
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With the worldwide spread of multi-drug resistant (MDR) bacteria, bacterial resistance has become a major issue affecting human health. Although traditional methods for obtaining antibiotics by screening bacterial strains have found the most available antibiotics for us, this method has resulted in fewer and fewer antibiotics in the past few decades and is increasingly difficult to find the new structure of the compound entity. At present, there are few drugs that can fight super-resistant bacteria in the clinic or even research. therefore, the development and application of new technologies to address the issue of bacterial resistance is imminent. Since the first bacterial genome was sequenced more than 20 years ago, a large number of bacterial genomic sequence information can provide clues for the discovery of new antibiotics. In this review, we briefly outline the available data sources and highlight the use of genomic mining and metagenomics in discovery of new antibiotics.
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This study investigated the occurrence and abundance of class 1 integrons and related antibiotic resistance genes (ARGs) in a sewage treatment plant (STP) of China. Totally, 189 bacterial strains were isolated from influent, activated sludge and effluent, and 40 isolates contained the integons with a complete structure. The intI1-carrying isolates were found to harbor two types of gene cassettes: dfr17-aadA5 and aadA2, conferring resistances to trimethoprim and streptomycin, which were further confirmed by antimicrobial susceptibility analysis. Many other gene cassettes were carried on integron, including qnrVC1, catB-8-blaoxa-10-aadA1-aac(6'), aadB-aacA29b, aadA2, aac(6')-1b, aadA6 and aadA12, which were detected using DNA cloning. Quantitative real time PCR showed that over 99% of the integrons was eliminated in activated sludge process, but average copy number of integrons in given bacterial cells was increased by 56% in treated sewage. Besides integrons, other mobile gene elements (MGEs) were present in the STP with high abundance. MGEs and the associated ARGs may be wide-spread in STPs, which constitute a potential hot spot for selection of antibiotic resistant bacteria and horizontal transfer of ARGs.
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BACKGROUND: The emergence of multidrug-resistant (MDR) Acinetobacter baumannii as an important opportunistic pathogen has given rise to significant therapeutic challenges in the treatment of nosocomial infections. In the present study, we assess the antibiotic resistance mechanisms of MDR A. baumannii strains by estimating the prevalence of antibiotic resistance determinants, including integrons, beta-lactamases, str genes, and gyrA and parC mutations. METHODS: Thirty-five MDR A. baumannii clinical isolates were collected from 3 Korean university hospitals over a 2-yr period. A. baumannii was confirmed by rpoB gene analysis. For each isolate, the minimal inhibitory concentrations (MICs) of 9 antibiotics were determined by the agar dilution method. PCR and DNA sequencing were used to identify the genes that potentially contribute to each resistance phenotype. RESULTS: Of the 35 MDR A. baumannii isolates examined, 7 antibiotic resistance gene determinants were detected. These resistance gene determinants included the gene bla(OXA-23), with an upstream element ISAba1 to promote increased gene expression and subsequent resistance to carbapenems, in 8 isolates (22.9%); aacA4, located within class 1 integrons, in 7 isolates (20.0%); and fluoroquinolone resistance conferred by gyrA and parC sense mutations in 31 isolates. CONCLUSIONS: Of the 35 MDR A. baumannii isolates, 26 (74.3%) from both outbreak and sporadic cases possessed at least 4 of the 7 antibiotic resistance gene determinants that give rise to the MDR phenotype. The co-occurrence of several resistance determinants may present a significant threat.
Subject(s)
Humans , Acinetobacter baumannii/drug effects , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Carbapenems/pharmacology , Drug Resistance, Multiple, Bacterial/genetics , Hospitals, University , Integrons/genetics , Microbial Sensitivity Tests , Republic of Korea , Sequence Analysis, DNAABSTRACT
OBJECTIVE To investigate the existence of genes for beta-lactam antibiotic resistance and for aminoglycosides modification enzymes(AMEs) in Pseudomonas aeruginosa(PAE) isolates from ICU patients and analyze the homology among strains.METHODS ?-Lactamase genes including TEM,SHV,OXA-10,PER,VEB,GES,CARB,IMP,VIM,SPM,GIM,DHA,FOX,MOX and oprD2,were detected by PCR amplication in 21 PAE isolates.The genes for AMES including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰwere determined by PCR amplification as well.RESULTS Among 21 isolates 21(100%),2(9.5%),1(4.8%),2(9.5%)and 4(19.0%) were positive for TEM,SHV,GES,CARB and VIM genes,respectively.The deletion of oprD2 gene was found in 14 out of 21 strains.Other ?-lactamase genes were absent in all isolates.As for AME genes,aac(3)-Ⅱ,aac(6″)-Ⅰ,aac(6)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aac(3)-Ⅰgenes were present in 19.0%,23.8%,9.5%,4.8%,and 19.0% of 21 isolates,However,aac(3)-Ⅰ gene was no position in any isolates.CONCLUSIONS P.aeruginosa carries various beta-lactamase and AME genes in ICU patients.Genetic cluster analysis suggested that clonal propagation result in nosocomial infection of PAE.
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The frequency of antibiotic resistance among Salmonella enterica serovar Typhimurium has increased due to the transfer of multiple resistance factors. We detected the 13 antibiotic resistance genes by multiplex-PCR and compared with the results of phage typing and antibiotic disk diffusion for 49 S. typhimurium isolated from food-poisoning outbreaks in Seoul from 1999 to 2002. Resistance genes for tetracycline, streptomycin, ampicillin, sulfonamide, amino-glycoside-modifying enzyme, chloramphenicol, kanamycin, and trimethoprim were detected in 67.3%, 57.1%, 26.5%, 8.1%, 8.1%, 5%, 2.0%, and 0% of isolates, respectively. Overall 28 isolates (57.1%) possessed two or more antibiotic resistance genes. Class 1 integron carrying multidrug resistace genes, ant(3")-IaB, blaPSE, qacE delta1/sul, and tet G were amplified especially in only DT104 isolates. Among the related resistance genes for same antibiotics, strA and strB for streptomycin resistance were simultaneously detected but tetA and tetB for tetracycline were sporadically detected. DT 104 isolates contained only aadA2 and tetG.