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Mechanisms of Resistance

Mechanisms of Increased Resistance to Chlorhexidine and Cross-Resistance to Colistin following Exposure of Klebsiella pneumoniae Clinical Isolates to Chlorhexidine

Matthew E. Wand, Lucy J. Bock, Laura C. Bonney, J. Mark Sutton
Matthew E. Wand
Public Health England, National Infection Service, Porton Down, Salisbury, Wiltshire, United Kingdom
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Lucy J. Bock
Public Health England, National Infection Service, Porton Down, Salisbury, Wiltshire, United Kingdom
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Laura C. Bonney
Public Health England, National Infection Service, Porton Down, Salisbury, Wiltshire, United Kingdom
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J. Mark Sutton
Public Health England, National Infection Service, Porton Down, Salisbury, Wiltshire, United Kingdom
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DOI: 10.1128/AAC.01162-16
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ABSTRACT

Klebsiella pneumoniae is an opportunistic pathogen that is often difficult to treat due to its multidrug resistance (MDR). We have previously shown that K. pneumoniae strains are able to “adapt” (become more resistant) to the widely used bisbiguanide antiseptic chlorhexidine. Here, we investigated the mechanisms responsible for and the phenotypic consequences of chlorhexidine adaptation, with particular reference to antibiotic cross-resistance. In five of six strains, adaptation to chlorhexidine also led to resistance to the last-resort antibiotic colistin. Here, we show that chlorhexidine adaptation is associated with mutations in the two-component regulator phoPQ and a putative Tet repressor gene (smvR) adjacent to the major facilitator superfamily (MFS) efflux pump gene, smvA. Upregulation of smvA (10- to 27-fold) was confirmed in smvR mutant strains, and this effect and the associated phenotype were suppressed when a wild-type copy of smvR was introduced on plasmid pACYC. Upregulation of phoPQ (5- to 15-fold) and phoPQ-regulated genes, pmrD (6- to 19-fold) and pmrK (18- to 64-fold), was confirmed in phoPQ mutant strains. In contrast, adaptation of K. pneumoniae to colistin did not result in increased chlorhexidine resistance despite the presence of mutations in phoQ and elevated phoPQ, pmrD, and pmrK transcript levels. Insertion of a plasmid containing phoPQ from chlorhexidine-adapted strains into wild-type K. pneumoniae resulted in elevated expression levels of phoPQ, pmrD, and pmrK and increased resistance to colistin, but not chlorhexidine. The potential risk of colistin resistance emerging in K. pneumoniae as a consequence of exposure to chlorhexidine has important clinical implications for infection prevention procedures.

FOOTNOTES

    • Received 3 June 2016.
    • Returned for modification 23 August 2016.
    • Accepted 9 October 2016.
    • Accepted manuscript posted online 31 October 2016.
  • Supplemental material for this article may be found at https://doi.org/10.1128/AAC.01162-16 .

  • © Crown copyright 2016.

The government of Australia, Canada, or the UK (“the Crown”) owns the copyright interests of authors who are government employees. The Crown Copyright is not transferable.

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Mechanisms of Increased Resistance to Chlorhexidine and Cross-Resistance to Colistin following Exposure of Klebsiella pneumoniae Clinical Isolates to Chlorhexidine
Matthew E. Wand, Lucy J. Bock, Laura C. Bonney, J. Mark Sutton
Antimicrobial Agents and Chemotherapy Dec 2016, 61 (1) e01162-16; DOI: 10.1128/AAC.01162-16

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Mechanisms of Increased Resistance to Chlorhexidine and Cross-Resistance to Colistin following Exposure of Klebsiella pneumoniae Clinical Isolates to Chlorhexidine
Matthew E. Wand, Lucy J. Bock, Laura C. Bonney, J. Mark Sutton
Antimicrobial Agents and Chemotherapy Dec 2016, 61 (1) e01162-16; DOI: 10.1128/AAC.01162-16
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KEYWORDS

Anti-Bacterial Agents
chlorhexidine
colistin
Klebsiella pneumoniae
Klebsiella pneumoniae
PhoPQ
chlorhexidine
colistin
smvA
smvR

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