Characterization of Staphylococcus epidermidis strains isolated from industrial cleanrooms under regular routine disinfection

J Appl Microbiol. 2017 May;122(5):1186-1196. doi: 10.1111/jam.13424. Epub 2017 Mar 29.

Abstract

Aims: The purpose of this study was the genotypic and phenotypic characterization of 57 strains of Staphylococcus epidermidis isolated from cleanroom environments, based on their biofilm formation and antimicrobial resistance profiles.

Methods and results: Biofilm formation was investigated using real-time PCR (icaA, aap, bhp genes), the Congo red agar method and the crystal violet assay. The majority of the strains (59·7%; 34/57) did not form biofilms according to the crystal violet assay, although the biofilm-associated genes were present in 94·7% (54/57) of the strains. Of the biofilm formers (40·4%; 23/57), 39·1% (9/23) have been identified as strong biofilm formers (>4× crystal violet absorbance cut-off). Resistance to a commercial disinfectant and its quaternary ammonium active component, didecyl-dimethyl-ammonium chloride (DDAC), was determined according to minimum inhibitory concentrations (MICs) and the presence of the qac (quaternary ammonium compound) genes. More than 95% (55/57) of the Staph. epidermidis strains had the qacA/B and qacC genes, but not the other qac genes. The MICs for the disinfectant and DDAC varied among the Staph. epidermidis strains, although none were resistant.

Conclusions: Although 59·6% of the Staph. epidermidis strains did not form biofilms and none were resistant to DDAC, more than 94% had the genetic basis for development of resistance to quaternary ammonium compounds, and among them at least 14·0% (8/57) might represent a high risk to cleanroom hygiene as strong biofim formers with qacA/B and qacC genes.

Significance and impact of the study: To assure controlled cleanroom environments, bacterial strains isolated from cleanroom environments need to be characterized regularly using several investigative methods.

Keywords: Staphylococcus epidermidis; biofilm; cleanroom; didecyl-dimethyl-ammonium chloride; quaternary ammonium compound; resistance.

MeSH terms

  • Biofilms / drug effects
  • Disinfectants / pharmacology*
  • Disinfection
  • Environment, Controlled
  • Humans
  • Industry / instrumentation
  • Industry / standards
  • Microbial Sensitivity Tests
  • Quaternary Ammonium Compounds / pharmacology
  • Staphylococcus epidermidis / classification
  • Staphylococcus epidermidis / drug effects
  • Staphylococcus epidermidis / genetics
  • Staphylococcus epidermidis / isolation & purification*

Substances

  • Disinfectants
  • Quaternary Ammonium Compounds