Step-Wise Increase in Tigecycline Resistance in Klebsiella pneumoniae Associated with Mutations in ramR, lon and rpsJ

Li, Fang; Chen, Qiong; Shi, Kezhang; Li, Xi; Shi, Qiucheng; He, Fang; Zhou, Jiancang; Yu, Yunsong; Hua, Xiaoting

doi:10.1371/journal.pone.0165019

Cited by 39 publications

(42 citation statements)

References 38 publications

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“…Deletion of the chromosomal β-lactamase of K. pneumoniae should allow clarification of whether piperacillin is expelled by pumps produced at normal levels. We found that AcrAB overproduction affects not only the activity of tigecycline, as previously described (Bialek-Davenet et al, 2015 ; Fang et al, 2016 ), but also that of ertapenem, piperacillin, ceftolozane, and temocillin. We also found that the MICs of ceftolozane+tazobactam were 4-fold lower than ceftolozane MICs in strains overproducing AcrAB.…”

Section: Discussionsupporting

confidence: 88%

Interplay Between Membrane Permeability and Enzymatic Barrier Leads to Antibiotic-Dependent Resistance in Klebsiella Pneumoniae

et al. 2018

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The interplay between membrane permeability alterations and the enzymatic barrier contributes to Klebsiella pneumoniae multidrug resistance. We assessed the specific effect of the efflux levels of the main efflux pumps (AcrAB and OqxAB), alone and associated with the loss of the main porins (OmpK35 and OMPK36), on the activity of various antibiotics by constructing a set of K. pneumoniae isogenic strains, including strains with plasmid-mediated β-lactamases (DHA-1, CTX-M-15, and OXA-48). The two pumps contributed to intrinsic chloramphenicol resistance and AcrAB to that of nalidixic acid and cefoxitin, whereas they had no impact on the activity of the other 11 antibiotics tested. We confirmed the expulsion of these three antibiotics by the two overproduced pumps and that of tigecycline by overproduced AcrAB, and showed that overproduced AcrAB also expelled ertapenem, piperacillin, ceftolozane, and ceftazidime. The sole loss of porins did not significantly affect the activity of the tested antibiotics, except ertapenem. The effect of efflux increases and porin loss on β-lactam activity was the highest in plasmid-mediated β-lactamase-producing strains. Thus, DHA-1-producing strains became non-susceptible (NS) to (i) ertapenem when there was an increase in efflux or porin loss, (ii) imipenem and ceftazidime+avibactam when the two mechanisms were associated, and (iii) temocillin when AcrAB was overproduced. The CTX-M-15-producing strains became NS to (i) ertapenem when there was no porin, (ii) ceftolozane+tazobactam when there was either overproduced OqxAB or porin loss, and (iii) temocillin when AcrAB was overproduced. OXA-48-producing strains known to be NS to temocillin were also NS to ceftolozane and they became NS to imipenem when the two pumps were overproduced or there was porin loss. Overall, this study shows that the balance between influx and efflux differentially modulates the activity of the tested antibiotics, an important point for evaluating the activity of future antibiotics or new combinations.

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Section: Discussionsupporting

confidence: 88%

Interplay Between Membrane Permeability and Enzymatic Barrier Leads to Antibiotic-Dependent Resistance in Klebsiella Pneumoniae

et al. 2018

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“…In order to investigate AcrAB-TolC efflux pump-dependent and pump-independent tigecycline resistance mechanisms, we analysed the sequences of the acrR , ramR , marR , soxR , lon , rpsJ and rpoC genes [ 25 , 26 ] by aligning our isolates with the reference genes of the tigecycline-susceptible isolate K. pneumoniae subsp. pneumoniae MGH78578 (GenBank Accession No.…”

Section: Resultsmentioning

confidence: 99%

“…The resistome of antimicrobial resistance genes was analysed using ResFinder-2.1 software (default identity thresholds [ID] 98%) provided by the Center for Genomic Epidemiology ( http://www.genomicepidemiology.org ) and the resources of the Pasteur MLST K. pneumoniae database, and by locally running the BLAST program. Previously described genes relating to AcrAB-TolC efflux pump-dependent and pump-independent tigecycline resistance mechanisms ( acrR , ramR , marR , soxR , lon , rpsJ and rpoC) [ 25 , 26 ], and chromosomal ( mgrB, pmrA/pmrB, phoP/phoQ ) and plasmidic ( mcr-1 and mcr-2 ) colistin resistance-related genes [ 27 , 28 ] were analysed in order to elucidate the possible determinants of tigecycline and colistin resistance. In the case of blaKPC -negative strains, the possible association between carbapenem resistance and the loss or alteration of outer membrane porins was investigated by studying the ompK35 and ompK36 genes.…”

Section: Methodsmentioning

confidence: 99%

Whole genome sequencing for the molecular characterization of carbapenem-resistant Klebsiella pneumoniae strains isolated at the Italian ASST Fatebenefratelli Sacco Hospital, 2012–2014

Rimoldi

Gentile²,

Pagani

et al. 2017

BMC Infect Dis

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BackgroundThe emergence of carbapenem-resistant Klebsiella pneumoniae strains is threatening antimicrobial treatment.MethodsSixty-eight carbapenemase-producing K. pneumoniae strains isolated at Luigi Sacco University Hospital-ASST Fatebenefratelli Sacco (Milan, Italy) between 2012 and 2014 were characterised microbiologically and molecularly. They were tested for drug susceptibility and carbapenemase phenotypes, investigated by means of repetitive extra-genic palindromic polymerase chain reaction (REP-PCR), and fully sequenced by means of next-generation sequencing for the in silico analysis of multi-locus sequence typing (MLST), their resistome, virulome and plasmid content, and their core single nucleotide polymorphism (SNP) genotypes.ResultsAll of the samples were resistant to carbapenems, other β-lactams and ciprofloxacin; many were resistant to aminoglycosides and tigecycline; and seven were resistant to colistin. Resistome analysis revealed the presence of blaKPC genes and, less frequently blaSHV, blaTEM, blaCTX-M and blaOXA, which are related to resistance to carbapenem and other β-lactams. Other genes conferring resistance to aminoglycoside, fluoroquinolone, phenicol, sulphonamide, tetracycline, trimethoprim and macrolide-lincosamide-streptogramin were also detected. Genes related to AcrAB-TolC efflux pump-dependent and pump-independent tigecycline resistance mechanisms were investigated, but it was not possible to clearly correlate the genomic features with tigecycline resistance because of the presence of a common mutation in susceptible, intermediate and resistant strains. Concerning colistin resistance, the mgrB gene was disrupted by an IS5-like element, and the mobile mcr-1 and mcr-2 genes were not detected in two cases. The virulome profile revealed type-3 fimbriae and iron uptake system genes, which are important during the colonisation stage in the mammalian host environment. The in silico detected plasmid replicons were classified as IncFIB(pQil), IncFIB(K), ColRNAI, IncX1, IncX3, IncFII(K), IncN, IncL/M(pMU407) and IncFIA(HI1). REP-PCR showed five major clusters, and MLST revealed six different sequence types: 512, 258, 307, 1519, 745 and 101. Core SNP genotyping, which led to four clusters, correlated with the MLST data. Isolates of the same sequencing type often had common genetic traits, but the SNP analysis allowed greater strain tracking and discrimination than either the REP-PCR or MLST analysis.ConclusionOur findings support the importance of implementing bacterial genomics in clinical medicine in order to complement traditional methods and overcome their limited resolution.Electronic supplementary materialThe online version of this article (10.1186/s12879-017-2760-7) contains supplementary material, which is available to authorized users.

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“…The overexpression of AcrAB-TolC and OqxAB in clinical K . pneumoniae isolates without a history of tigecycline exposure may be due to exposure to other antimicrobials that are transported by the same efflux pumps 8 – 12 . The eravacycline-resistant and eravacycline-heteroresistant K .…”

Section: Discussionmentioning

confidence: 99%

“…Meanwhile, mutations in ramR , acrR , and rpsJ genes have also been reported to contribute to K . pneumoniae resistance to tigecycline 9 , 12 .…”

Section: Introductionmentioning

confidence: 99%

Overexpression of OqxAB and MacAB efflux pumps contributes to eravacycline resistance and heteroresistance in clinical isolates of Klebsiella pneumoniae

Zheng

Wang

Sun

et al. 2018

Emerging Microbes & Infections

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This study investigated the characteristics and mechanisms of eravacycline resistance and heteroresistance in clinical Klebsiella pneumoniae isolates. A total of 393 clinical K. pneumoniae isolates were collected and subjected to eravacycline and tigecycline MIC determinations using the agar dilution method. Eravacycline heteroresistance was assessed by a population analysis profile (PAP). The expression levels of efflux pumps and their regulators were determined by quantitative reverse-transcription PCR (qRT-PCR). This study identified 67 eravacycline-nonsusceptible isolates; among the extended-spectrum β-lactamase (ESBL)-positive isolates, eravacycline-nonsusceptible isolates were detected more frequently than tigecycline-nonsusceptible isolates (21.7% vs. 9.4%, p = 0.001). The study sample was observed to include 20 K. pneumoniae isolates with eravacycline heteroresistance. Compared to the reference strain, oqxA or oqxB overexpression was observed in nine eravacycline-nonsusceptible isolates (range, 35.64–309.02-fold) and 13 eravacycline-heteroresistant isolates (8.42–296.34-fold). The overexpression of macA or macB was detected in 12 eravacycline-heteroresistant isolates (3.23–28.35-fold). Overexpression of the efflux pump regulator gene ramA was observed in 11 eravacycline-nonsusceptible isolates (3.33–94.05-fold) and 18 eravacycline-heteroresistant isolates (3.89–571.70-fold). The eravacycline MICs were increased by one–fourfold by overexpression of oqxAB or macAB in three eravacycline-sensitive isolates. In conclusion, the overexpression of OqxAB and MacAB efflux pumps and the transcriptional regulator RamA were suggested to be involved in K. pneumoniae eravacycline resistance and heteroresistance.

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Step-Wise Increase in Tigecycline Resistance in Klebsiella pneumoniae Associated with Mutations in ramR, lon and rpsJ

Cited by 39 publications

References 38 publications

Interplay Between Membrane Permeability and Enzymatic Barrier Leads to Antibiotic-Dependent Resistance in Klebsiella Pneumoniae

Interplay Between Membrane Permeability and Enzymatic Barrier Leads to Antibiotic-Dependent Resistance in Klebsiella Pneumoniae

Whole genome sequencing for the molecular characterization of carbapenem-resistant Klebsiella pneumoniae strains isolated at the Italian ASST Fatebenefratelli Sacco Hospital, 2012–2014

Overexpression of OqxAB and MacAB efflux pumps contributes to eravacycline resistance and heteroresistance in clinical isolates of Klebsiella pneumoniae

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