Piperacillin-Tazobactam-Resistant/Third-Generation Cephalosporin-Susceptible Escherichia coli and Klebsiella pneumoniae Isolates: Resistance Mechanisms and In vitro-In vivo Discordance

Int J Antimicrob Agents. 2020 Mar;55(3):105885. doi: 10.1016/j.ijantimicag.2020.105885. Epub 2020 Jan 8.

Abstract

We previously reported the detection of Escherichia coli and Klebsiella pneumoniae that displayed in vitro piperacillin-tazobactam (TZP) resistance but were susceptible to third-generation cephalosporins (TZP-R/Ceph3-S). In this study, we assessed the phenotypic and genotypic profiles of 12 clinical non-clonal TZP-R/Ceph3-S E. coli and K. pneumoniae isolates derived from bloodstream infections. Whole-genome sequencing revealed that most of the TZP-R/Ceph3-S E. coli and K. pneumoniae isolates examined harbored blaTEM-1 and blaSHV-1 genes, respectively, but none harbored extended-spectrum β-lactamase, AmpC β-lactamase or carbapenemase genes. Increasing the tazobactam concentration from 4 mg/L to 16 mg/L restored TZP in vitro susceptibility among E. coli isolates expressing TEM-1, but had minimal impact on the susceptibility of K. pneumoniae to TZP. Real-time qPCR analysis showed that blaTEM-1 expression was amplified in TZP-R E. coli upon incubation with sub-inhibitory TZP concentrations. Using an immunocompetent murine septicemia model, the efficacy of TZP against TZP-R/Ceph3-S isolates was assessed using TZP doses that mimicked human plasma exposures following intravenous (IV) administration of TZP 4.5 g q6h over 0.5 h for 24 h. Efficacy was assessed by survival through 96 h. There was high mortality in untreated control mice for all tested isolates. Compared with controls, TZP human-simulated exposure significantly improved survival for all TZP-R/Ceph3-S E. coli and K. pneumoniae isolates examined (P < 0.05). Thus, TZP was associated with remarkable in vivo activity against TZP-R/Ceph3-S E. coli and K. pneumoniae despite the observed resistance in vitro.

Keywords: Bacteremia; Piperacillin-Tazobactam; Septicemia model; TEM-1.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Cephalosporins / pharmacology*
  • Cephalosporins / therapeutic use
  • Drug Resistance, Bacterial / drug effects
  • Escherichia coli / drug effects*
  • Escherichia coli / genetics
  • Escherichia coli Infections / drug therapy*
  • Humans
  • Klebsiella Infections / drug therapy*
  • Klebsiella pneumoniae / drug effects*
  • Klebsiella pneumoniae / genetics
  • Mice
  • Piperacillin, Tazobactam Drug Combination / pharmacology*
  • Piperacillin, Tazobactam Drug Combination / therapeutic use
  • beta-Lactamases / genetics

Substances

  • Bacterial Proteins
  • Cephalosporins
  • Piperacillin, Tazobactam Drug Combination
  • AmpC beta-lactamases
  • beta-Lactamases
  • carbapenemase