Optimizing a Screening Protocol for Potential Extended-Spectrum β-Lactamase Escherichia coli on MacConkey Agar for Use in a Global Surveillance Program

J Clin Microbiol. 2020 Aug 24;58(9):e01039-19. doi: 10.1128/JCM.01039-19. Print 2020 Aug 24.

Abstract

The increasing prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli is worrisome. Coordinated efforts to better understand global prevalence and risk factors are needed. Developing lower- and middle-income countries need reliable, readily available, and cost-effective solutions for detecting ESBL E. coli to contribute to global surveillance. We evaluated MacConkey agar supplemented with ceftriaxone or cefotaxime as a screening method for accurately detecting and quantifying potential ESBL E. coli MacConkey agar from eight manufacturers, representing seven countries, was prepared with 2 or 4 μg/ml ceftriaxone or cefotaxime. Four E. coli strains (NC11, ATCC 25922, CM-13457, and CM-10455) and one Klebsiella pneumoniae strain (CM-11073) were grown overnight, serially diluted, and plated in triplicate for enumeration on all medium combinations. After recovery was assessed, US-1 MacConkey agar with cefotaxime was used to further evaluate the reproducibility and detection of potential ESBL E. coli from poultry cecal (n = 30) and water (n = 30) samples. Results indicated the recovery of E. coli 13457 from four MacConkey agar manufacturers was reduced by up to 4 log CFU/ml, and phenotypic differences in colony size and color were apparent for each manufacturer for control E. coli strains. A true ESBL, NC11, was not reduced with 4 μg/ml cefotaxime. From ceca and water, potential ESBL E. coli isolates were only confirmed from MacConkey agar with 4 μg/ml cefotaxime, where 45% and 16.6% of E. coli isolates phenotypically expressed ESBL production. The quality and reproducibility of MacConkey agar varied by manufacturer, which suggests that a single manufacturer and medium type should be selected for global monitoring efforts so that training and interpretation can be standardized.

Keywords: ESBL; Escherichia coli; MacConkey agar; global surveillance.

MeSH terms

  • Agar
  • Anti-Bacterial Agents / pharmacology
  • Escherichia coli Infections* / epidemiology
  • Escherichia coli*
  • Humans
  • Klebsiella pneumoniae
  • Microbial Sensitivity Tests
  • Reproducibility of Results
  • beta-Lactamases

Substances

  • Anti-Bacterial Agents
  • Agar
  • beta-Lactamases