Phenotypic variants of staphylococci and their underlying population distributions following exposure to stress

PLoS One. 2013 Oct 18;8(10):e77614. doi: 10.1371/journal.pone.0077614. eCollection 2013.

Abstract

This study investigated whether alterations in environmental conditions would induce the formation of small colony variant phenotypes (SCV) with associated changes in cell morphology and ultra-structure in S. aureus, s. epidermidis, and S. lugdunensis. Wild-type clinical isolates were exposed to low temperature (4 °C), antibiotic stress (penicillin G and vancomycin; 0-10,000 µg mL(-1)), pH stress (pH 3-9) and osmotic challenge (NaCl concentrations of 0-20%). Changes in cell diameter, cell-wall thickness, and population distribution changes (n ≥ 300) were assessed via scanning and transmission electron microscopy (SEM and TEM), and compared to control populations. Our analyses found that prolonged exposure to all treatments resulted in the subsequent formation of SCV phenotypes. Observed SCVs manifested as minute colonies with reduced haemolysis and pigmentation (NaCl, pH and 4°C treatments), or complete lack thereof (antibiotic treatments). SEM comparison analyses revealed significantly smaller cell sizes for SCV populations except in S. aureus and S. epidermidis 10% NaCl, and S. epidermidis 4 °C (p<0.05). Shifts in population distribution patterns were also observed with distinct sub-populations of smaller cells appearing for S. epidermidis, and S. lugdunensis. TEM analyses revealed significantly thicker cell-walls in all treatments and species except S. lugdunensis exposed to 4 °C. These findings suggest that staphylococci adapted to environmental stresses by altering their cell size and wall thickness which could represent the formation of altered phenotypes which facilitate survival under harsh conditions. The phenotypic response was governed by the type of prevailing environmental stress regime leading to appropriate alterations in ultra-structure and size, suggesting downstream changes in gene expression, the proteome, and metabolome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / adverse effects
  • Cell Size
  • Cell Wall / physiology
  • Cell Wall / ultrastructure
  • Environment
  • Hydrogen-Ion Concentration
  • Osmotic Pressure / physiology
  • Phenotype
  • Staphylococcus / drug effects
  • Staphylococcus / physiology*
  • Staphylococcus / ultrastructure*
  • Stress, Physiological / physiology*
  • Temperature

Substances

  • Anti-Bacterial Agents

Grants and funding

This work was partially supported via a University of Newcastle International Scholarship and Gideon Lang Scholarship, and additional funding from the Harold Stannet Williams and Judith Mason Research Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.