Characterization of the genetic environment of blaKPC in Escherichia coli isolates from hospitals in China

ABSTRACT Carbapenem resistance in Enterobacteriaceae members has become a major challenge, and the genetic environment of blaKPC, encoding Klebsiella pneumoniae carbapenemases, has not been fully clarified in China. In this study, we aimed to explore the genetic environment of blaKPC in 25 carbapenem-resistant E. coli isolates from hospitals in Hangzhou Province, China. Antimicrobial susceptibility against 22 common antimicrobial agents was tested. Polymerase chain reaction (PCR) analysis was performed for screening of the resistent genes, such as blaKPC, blaCTX-M, blaTEM, blaSHV, blaNDM, qnrA, qnrB, qnrS, aac(6’)-Ib, armA and rmtB. The genetic environment of blaKPC were determinedin one isolate. blaKPC was detected by PCR in all the clinical E. coli isolates. There were no strains carrying blaNDM, qnrA and armA. The genetic environment of blaKPC showed that blaKPC dissemination is plasmid mediated and that it is located in the Tn3–Tn4401 transposon complex. Encoding of blaKPC-2 was responsible for carbapenem resistance in the 25 E. coli isolates. The genetic environment of blaKPC was characterized by the Tn3–Tn4401 complex. Our findings may provide a theoretical basis for clinical drug-resistance monitoring, anti-infection treatment and hospital infection control.

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Characterization of Carbapenem-Resistant Enterobacteriaceae Cultured From Retail Meat Products, Patients, and Porcine Excrement in China

Frontiers in Microbiology ◽

10.3389/fmicb.2021.743468 ◽

2021 ◽

Vol 12 ◽

Author(s):

Jie Feng ◽

Qian Xiang ◽

Jiangang Ma ◽

Pei Zhang ◽

Kun Li ◽

...

Keyword(s):

Meat Products ◽

Carbapenem Resistance ◽

Multidrug Resistant ◽

E Coli ◽

Carbapenem Resistant ◽

Retail Meat ◽

Different Origins ◽

Different Sources ◽

Carbapenem Resistant Enterobacteriaceae

The emergence and dissemination of carbapenem-resistant Enterobacteriaceae (CRE) is a growing concern to animal and public health. However, little is known about the spread of CRE in food and livestock and its potential transmission to humans. To identify CRE strains from different origins and sources, 53 isolates were cultured from 760 samples including retail meat products, patients, and porcine excrement. Antimicrobial susceptibility testing was carried out, followed by phylogenetic typing, whole-genome sequencing, broth mating assays, and plasmids analyses. Forty-three Escherichia coli, nine Klebsiella pneumoniae, and one Enterobacter cloacae isolates were identified, each exhibiting multidrug-resistant phenotypes. Genetically, the main sequence types (STs) of E. coli were ST156 (n = 7), ST354 (n = 7), and ST48 (n = 7), and the dominant ST of K. pneumoniae is ST11 (n = 5). blaNDM–5 (n = 40) of E. coli and blaKPC–2 (n = 5) were the key genes that conferred carbapenem resistance phenotypes in these CRE strains. Additionally, the mcr-1 gene was identified in 17 blaNDM-producing isolates. The blaNDM–5 gene from eight strains could be transferred to the recipients via conjugation assays. Two mcr-1 genes in the E. coli isolates could be co-transferred along with the blaNDM–5 genes. IncF and IncX3 plasmids have been found to be predominantly associated with blaNDM gene in these strains. Strains isolated in our study from different sources and regions tend to be concordant and overlap. CRE strains from retail meat products are a reservoir for transition of CRE strains between animals and humans. These data also provide evidence of the dissemination of CRE strains and carbapenem-resistant genes between animal and human sources.

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Molecular Characterization of Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates from Egyptian Patients

10.21203/rs.3.rs-123397/v1 ◽

2020 ◽

Author(s):

Reem M Hassan ◽

Sherifa T Salem ◽

Saly Ismail Mostafa Hassan ◽

Asmaa Hegab ◽

Yasmine S Elkholy

Keyword(s):

Acinetobacter Baumannii ◽

Molecular Characterization ◽

Antimicrobial Agents ◽

Treatment Options ◽

Carbapenem Resistance ◽

Common Mechanism ◽

Carbapenem Resistant ◽

Egyptian Patients ◽

Global Threat

Abstract Acinetobacter baumannii (A. baumannii) represents a global threat owing to its ability to resist most of the currently available antimicrobial agents. Moreover, emergence of carbapenem resistant A. baumannii (CR-AB) isolates limits the available treatment options. Enzymatic degradation by variety of ß-lactamases, have been identified as the most common mechanism of carbapenem resistance in A. baumannii. The alarming increase in the prevalence of CR-AB necessitates continuous screening and molecular characterization to appreciate the problem. The present study was performed to assess the prevalence and characterize carbapenemases among 206 CR-AB isolated from various clinical specimens collected from different intensive care units at Kasr Al-Aini Hospital. All isolates were confirmed to be A. baumannii by detection of the blaOXA-51-like gene. Molecular screening of 13 common Ambler class bla carbapenemases genes in addition to insertion sequence (IS-1) upstream OXA-23 was performed by using four sets of multiplex PCR, followed by identification using gene sequencing technology. Among the investigated genes, the prevalence of blaOXA-23, and blaOXA-58 were 77.7%, and 1.9%, respectively. The ISAba1 was detected in 10% of the blaOXA-23 positive isolates. The prevalence of metallo-β-lactamases (MBLs) studied; blaNDM-1, blaSPM, blaVIM, blaSIM-1 were 11.7%, 6.3%, 0.5%, and 0.5% respectively. One of class A; bla KPC was detected in 10.7% of the investigated isolates. blaOXA-24/40, blaIMP, blaGES, blaVEB and blaGIM were not detected in any of the studied isolates. Moreover, 18.4% of the isolates have shown to harbor two or more of the screened bla genes. We concluded that the most prevalent type of ß-lactamases genes among CR-AB isolates collected from Egyptian patients were blaOXA-23 followed by blaNDM-1 and blaKPC.

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Arising Prevalence of OXA-48 producer Escherichia coli and OXA-48 with NDM co-producer Klebsiella pneumoniae Strains

Revista Romana de Medicina de Laborator ◽

10.2478/rrlm-2019-0030 ◽

2019 ◽

Vol 27 (3) ◽

pp. 319-326 ◽

Cited By ~ 2

Author(s):

Aylin Uskudar-Guclu ◽

Mustafa Guney ◽

Ali Korhan Sig ◽

Selcuk Kilic ◽

Mehmet Baysallar

Keyword(s):

Antimicrobial Agents ◽

Carbapenem Resistance ◽

Resistance Mechanisms ◽

Resistant Isolate ◽

Diffusion Method ◽

Clinical Samples ◽

Disc Diffusion ◽

E Coli ◽

Carbapenem Resistant ◽

Resistance Rates

Abstract Background/aim: This prospective study aimed to determine the presence of the most common carbapenemase genes, blaOXA-48, blaKPC, blaIMP, blaVIM and blaNDM on carbapenem resistant clinical K.pneumoniae and E.coli isolates. Materials and methods: Isolates were selected according to EUCAST guideline; gradient test and disc diffusion with both meropenem and ertapenem discs. Resistance rates of these isolates to other antimicrobial agents were also examined by disc diffusion method. Carbapenem resistance gene were investigated by using Real-Time PCR. Results: A total of 3845 E. coli and 1689 K.pneumoniae isolates from clinical samples between January 2015 and April 2017 were evaluated. The 419 isolates were found as carbapenem resistant but only the first resistant isolate (n=155; 126 K.pneumoniae and 29 E.coli) of each patient were included. Carbapenem resistant isolates were most frequently isolated from intensive care units (48.8%). Colistin was the most effective antibiotic (91.0%). The 121 (78.1%) of the tested isolates were positive for OXA-48 (103 K.pneumoniae and 18 E.coli) and 9 K. pneumoniae carrying blaNDM were also positive for blaOXA-48. VIM, IMP and KPC type carbapenemases were not detected in any isolates. Conclusion: Carbapenem-resistant pathogens have been shown to be able to develop resistance mechanisms with more than one carbapenemase encoding gene.

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Synergistic Activity of Equol and Meropenem against Carbapenem-Resistant Escherichia coli

Antibiotics ◽

10.3390/antibiotics10020161 ◽

2021 ◽

Vol 10 (2) ◽

pp. 161

Author(s):

Hye-Rim Kim ◽

Yong-Bin Eom

Keyword(s):

Escherichia Coli ◽

Synergistic Effect ◽

Bacterial Infections ◽

Treatment Options ◽

Carbapenem Resistance ◽

Bacterial Biofilm ◽

Pcr Analysis ◽

Synergistic Activity ◽

E Coli ◽

Carbapenem Resistant

The emergence of carbapenem-resistant Enterobacterales (CRE) seriously limits treatment options for bacterial infections. Combined drugs are an effective strategy to treat these resistant strains. This study aimed to evaluate the synergistic effect of equol and meropenem against carbapenem-resistant Escherichia coli. First, this study investigated the antibacterial activity of carbapenems on clinically isolated E. coli strains by analyzing the minimum inhibitory concentrations (MICs). The E. coli strains were all resistant to carbapenem antibiotics. Therefore, we confirmed the cause of carbapenem resistance by detecting blaKPC and blaOXA-48 among the carbapenemase genes using polymerase chain reaction (PCR) analysis. Checkerboard and time-kill analyses confirmed that equol restored the susceptibility of carbapenem-resistant E. coli to meropenem. Also, the transcription levels of specific carbapenemase genes in E. coli were significantly suppressed by equol. The study also evaluated the anti-virulence effects of equol on bacterial biofilm and motility through phenotypic and genotypic analyses. In conclusion, our results revealed that equol had a synergistic effect with meropenem on carbapenem-resistant E. coli. Therefore, this study suggests that equol is a promising antibiotic adjuvant that prevents the expression of carbapenemases and virulence factors in carbapenem-resistant E. coli.

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Molecular characterization of carbapenem-resistant Acinetobacter baumannii clinical isolates from Egyptian patients

PLoS ONE ◽

10.1371/journal.pone.0251508 ◽

2021 ◽

Vol 16 (6) ◽

pp. e0251508

Author(s):

Reem M. Hassan ◽

Sherifa T. Salem ◽

Saly Ismail Mostafa Hassan ◽

Asmaa Sayed Hegab ◽

Yasmine S. Elkholy

Keyword(s):

Acinetobacter Baumannii ◽

Molecular Characterization ◽

Antimicrobial Agents ◽

Treatment Options ◽

Carbapenem Resistance ◽

Common Mechanism ◽

Carbapenem Resistant ◽

Egyptian Patients ◽

Global Threat

Acinetobacter baumannii (A. baumannii) represents a global threat owing to its ability to resist most of the currently available antimicrobial agents. Moreover, emergence of carbapenem resistant A. baumannii (CR-AB) isolates limits the available treatment options. Enzymatic degradation by variety of ß-lactamases, have been identified as the most common mechanism of carbapenem resistance in A. baumannii. The alarming increase in the prevalence of CR-AB necessitates continuous screening and molecular characterization to appreciate the problem. The present study was performed to assess the prevalence and characterize carbapenemases among 206 CR-AB isolated from various clinical specimens collected from different intensive care units at Kasr Al-Aini Hospital. All isolates were confirmed to be A. baumannii by detection of the blaOXA-51-like gene. Molecular screening of 13 common Ambler class bla carbapenemases genes in addition to insertion sequence (IS-1) upstream OXA-23 were performed by using four sets of multiplex PCR, followed by identification using gene sequencing technology. Among the investigated genes, the prevalence of blaOXA-23, and blaOXA-58 were 77.7%, and 1.9%, respectively. The ISAba1 was detected in 10% of the blaOXA-23 positive isolates. The prevalence of metallo-β-lactamases (MBLs) studied; blaNDM-1, blaSPM, blaVIM, blaSIM-1 were 11.7%, 6.3%, 0.5%, and 0.5% respectively. One of class A; bla KPC was detected in 10.7% of the investigated isolates. blaOXA-24/40, blaIMP, blaGES, blaVEB and blaGIM were not detected in any of the studied isolates. Moreover, 18.4% of the isolates have shown to harbor two or more of the screened bla genes. We concluded that the most prevalent type of ß-lactamases genes among CR-AB isolates collected from Egyptian patients were blaOXA-23 followed by blaNDM-1 and blaKPC.

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Genomic and Molecular Characterization of Clinical Isolates of Enterobacteriaceae Harboring mcr-1 in Colombia, 2002 to 2016

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00841-17 ◽

2017 ◽

Vol 61 (12) ◽

Cited By ~ 28

Author(s):

Sandra Yamile Saavedra ◽

Lorena Diaz ◽

Magdalena Wiesner ◽

Adriana Correa ◽

Stefany Alejandra Arévalo ◽

...

Keyword(s):

Antimicrobial Agents ◽

Clinical Isolates ◽

Gram Negative ◽

Content Type ◽

E Coli ◽

Carbapenem Resistant ◽

Serovar Typhimurium ◽

Chromosome Conjugation ◽

Sequence Types

ABSTRACT Polymyxins are last-resort antimicrobial agents used to treat infections caused by carbapenem-resistant Enterobacteriaceae. Due to the worldwide dissemination of polymyxin resistance in animal and human isolates, we aimed to characterize polymyxin resistance associated with the presence of mcr-1 in Enterobacteriaceae and nonfermenter Gram-negative bacilli, using isolates collected retrospectively in Colombia from 2002 to 2016. A total of 5,887 Gram-negative clinical isolates were studied, and 513 were found to be resistant to the polymyxins. Susceptibility to colistin was confirmed by broth microdilution for all mcr-1-positive isolates, and these were further subjected to whole-genome sequencing (WGS). The localization of mcr-1 was confirmed by S1 pulsed-field gel electrophoresis (S1-PFGE) and CeuI-PFGE hybridization. Transferability was evaluated by mating assays. A total of 12 colistin-resistant isolates recovered after 2013 harbored mcr-1, including 8 Escherichia coli, 3 Salmonella enterica serovar Typhimurium, and 1 Klebsiella pneumoniae isolate. E. coli isolates were unrelated by PFGE and belonged to 7 different sequence types (STs) and phylogroups. S. Typhimurium and K. pneumoniae isolates belonged to ST34 and ST307, respectively. The mcr-1 gene was plasmid borne in all isolates but two E. coli isolates which harbored it on the chromosome. Conjugation of mcr-1 was successful in 8 of 10 isolates (8.2 × 10−5 to 2.07 × 10−1 cell per recipient). Plasmid sequences showed that the mcr-1 plasmids belonged to four different Inc groups (a new IncP-1 variant and the IncFII, IncHI1, and IncH families). Our results indicate that mcr-1 is circulating in clinical isolates of colistin-resistant Enterobacteriaceae in Colombia and is mainly harbored in transferable plasmids.

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Characterization of fosfomycin resistance and molecular epidemiology among carbapenem-resistant Klebsiella pneumoniae strains from two tertiary hospitals in China

BMC Microbiology ◽

10.1186/s12866-021-02165-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Haichen Wang ◽

Changhang Min ◽

Jun Li ◽

Ting Yu ◽

Yongmei Hu ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Molecular Mechanisms ◽

Genomic Analysis ◽

Carbapenem Resistance ◽

E Coli ◽

Carbapenem Resistant ◽

Its Gene ◽

Tertiary Hospitals ◽

Resistant Strains

Abstract Background Fosfomycin has been proven to be a vital choice to treat infection caused by multidrug resistance bacteria, especially carbapenem-resistant Klebsiella pneumoniae (CRKP). However, fosfomycin resistant cases has been reported gradually. In this study, we reported the fosfomycin-resistant rate in CRKP strains and further revealed the molecular mechanisms in resistance gene dissemination. Results A total of 294 non-duplicated CRKP strains were collected. And 55 fosfomyin-resistant strains were detected, 94.5% of which were clustered to sequence type (ST) 11 by PCR followed up sequencing. PFGE further revealed two major groups and four singletons. The positive rates of genes responsible to fosfomycin and carbapenem resistance were 81.8% (fosA3), 12.7% (fosA5) and 94.5% (blaKPC-2), respectively. Genomic analysis confirmed insertion sequence (IS) 26 was the predominant structure surrounding fosA3. The fosA3 genes in six isolates were located on plasmids which were able to transfer to E. coli J53 recipient cells by means of conjugation. Conclusions Although the resistant rate of CRKP to fosfomycin is relatively low in our area, considering its gene is located on transferrable plasmid and inserted in IS structure, continuous monitoring is still needed.

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Molecular characterization of clinical carbapenem-resistant Enterobacterales from Qatar

European Journal of Clinical Microbiology & Infectious Diseases ◽

10.1007/s10096-021-04185-7 ◽

2021 ◽

Author(s):

Fatma Ben Abid ◽

Clement K. M. Tsui ◽

Yohei Doi ◽

Anand Deshmukh ◽

Christi L. McElheny ◽

...

Keyword(s):

Common Species ◽

Clinical Samples ◽

Whole Genome ◽

E Coli ◽

Carbapenem Resistant ◽

Genes Encoding ◽

Encoding Genes ◽

Sequence Types ◽

Common Sequence

AbstractOne hundred forty-nine carbapenem-resistant Enterobacterales from clinical samples obtained between April 2014 and November 2017 were subjected to whole genome sequencing and multi-locus sequence typing. Klebsiella pneumoniae (81, 54.4%) and Escherichia coli (38, 25.5%) were the most common species. Genes encoding metallo-β-lactamases were detected in 68 (45.8%) isolates, and OXA-48-like enzymes in 60 (40.3%). blaNDM-1 (45; 30.2%) and blaOXA-48 (29; 19.5%) were the most frequent. KPC-encoding genes were identified in 5 (3.6%) isolates. Most common sequence types were E. coli ST410 (8; 21.1%) and ST38 (7; 18.4%), and K. pneumoniae ST147 (13; 16%) and ST231 (7; 8.6%).

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Characterization of resistance mechanisms of Enterobacter cloacae Complex co-resistant to carbapenem and colistin

BMC Microbiology ◽

10.1186/s12866-021-02250-x ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Shixing Liu ◽

Renchi Fang ◽

Ying Zhang ◽

Lijiang Chen ◽

Na Huang ◽

...

Keyword(s):

Lipid A ◽

Efflux Pump ◽

Resistance Mechanism ◽

Carbapenem Resistance ◽

Enterobacter Cloacae ◽

Resistance Mechanisms ◽

Colistin Resistance ◽

Carbapenem Resistant ◽

Horizontal Spread

Abstract Background The emergence of carbapenem-resistant and colistin-resistant ECC pose a huge challenge to infection control. The purpose of this study was to clarify the mechanism of the carbapenems and colistin co-resistance in Enterobacter cloacae Complex (ECC) strains. Results This study showed that the mechanisms of carbapenem resistance in this study are: 1. Generating carbapenemase (7 of 19); 2. The production of AmpC or ESBLs combined with decreased expression of out membrane protein (12 of 19). hsp60 sequence analysis suggested 10 of 19 the strains belong to colistin hetero-resistant clusters and the mechanism of colistin resistance is increasing expression of acrA in the efflux pump AcrAB-TolC alone (18 of 19) or accompanied by a decrease of affinity between colistin and outer membrane caused by the modification of lipid A (14 of 19). Moreover, an ECC strain co-harboring plasmid-mediated mcr-4.3 and blaNDM-1 has been found. Conclusions This study suggested that there is no overlap between the resistance mechanism of co-resistant ECC strains to carbapenem and colistin. However, the emergence of strain co-harboring plasmid-mediated resistance genes indicated that ECC is a potential carrier for the horizontal spread of carbapenems and colistin resistance.

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Conditions to Prolonged Release of Microencapsulated Carvacrol on Alginate Films as Affected by Emulsifier Type and PH

International Journal of Polymer Science ◽

10.1155/2015/173193 ◽

2015 ◽

Vol 2015 ◽

pp. 1-12 ◽

Cited By ~ 2

Author(s):

Silvia Matiacevich ◽

Natalia Riquelme ◽

María Lidia Herrera

Keyword(s):

Storage Time ◽

Antimicrobial Agents ◽

Antimicrobial Properties ◽

Edible Film ◽

Initial Time ◽

Tween 20 ◽

Prolonged Release ◽

E Coli ◽

Fresh Foods

Alginate from algal biomass is used as edible film and the incorporation of antimicrobial agents improves its performance to increase the shelf-life of fresh foods. However, environmental conditions and intrinsic properties of films influence their release. The aim of this study was to investigate the effect of the concentration and type of encapsulating agent and pH of emulsions on the physical and antimicrobial properties of alginate-carvacrol films. Films containing alginate, carvacrol as antimicrobial agent, and Tween 20 or trehalose (0.25 and 0.75% w/w) as encapsulating agents were obtained from suspensions at pH 4 and pH 8. Physical characterization of emulsions and films and antimicrobial properties (E. coliandB. cinerea) was evaluated. Results showed that droplets size depended on trehalose concentration, but emulsion stability depended on pH and type of encapsulating agent, being more stable samples with trehalose at pH 4. Although films with Tween 20 presented the highest opacity, they showed the best antimicrobial properties at initial time; however, during storage time, they lost their activity before samples with trehalose and relative humidity (RH) was the principal factor to influence their release. Therefore, sample formulated with 0.25% trehalose at pH 4 and stored at 75% RH had the best potential as edible film for fresh fruits.

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