The progress of type II persisters of Escherichia coli O157:H7 to a non-culturable state during prolonged exposure to antibiotic stress with revival being aided through acid-shock treatment and provision of methyl pyruvate

Can J Microbiol. 2021 Jul;67(7):518-528. doi: 10.1139/cjm-2020-0339. Epub 2020 Oct 30.

Abstract

Persisters are a form of dormancy in bacteria that provide temporary resistance to antibiotics. The following reports on the formation of Escherichia coli O157:H7 E318 type II persisters from a protracted (8 days) challenge with ampicillin. Escherichia coli O157:H7 followed a multiphasic die-off pattern with an initial rapid decline (Phase I) of susceptible cells that transitioned to a slower rate representing tolerant cells (Phase II). After 24 h post-antibiotic challenge, the E. coli O157:H7 levels remained relatively constant at 2 log CFU/mL (Phase III), but became non-culturable within 8-days (Phase IV). The revival of persisters in Phase III could be achieved by the removal of antibiotic stress, although those in Phase IV required an extended incubation period or application of acid-shock. The carbon utilization profile of persister cells was less diverse compared with non-persisters, with only methyl pyruvate being utilized from the range tested. Inclusion of methyl pyruvate in tryptic soy agar revived non-cultural persisters, presumably by stimulating metabolism. The results suggest that persisters could be subdivided into culturable or non-culturable cells, with the former representing a transition state to the latter. The study provided insights into how to revive cells from dormancy to aid enumeration and control.

Keywords: Escherichia coli O157H7; acid shock; ampicillin; ampicilline; antibiotic resistance; choc acide; ester de pyruvate de méthyle; methyl pyruvate ester; persisters; résistance aux antibiotiques.

MeSH terms

  • Acids / pharmacology*
  • Anti-Bacterial Agents / pharmacology*
  • Colony Count, Microbial
  • Escherichia coli O157 / drug effects*
  • Escherichia coli O157 / genetics
  • Escherichia coli O157 / growth & development*
  • Pyruvates / pharmacology*

Substances

  • Acids
  • Anti-Bacterial Agents
  • Pyruvates
  • methyl pyruvate