Studies of Antibiotic Resistance of Beta-Lactamase Bacteria under Different Nutrition Limitations at the Single-Cell Level

PLoS One. 2015 May 20;10(5):e0127115. doi: 10.1371/journal.pone.0127115. eCollection 2015.

Abstract

Drug resistance involves many biological processes, including cell growth, cell communication, and cell cooperation. In the last few decades, bacterial drug resistance studies have made substantial progress. However, a major limitation of the traditional resistance study still exists: most of the studies have concentrated on the average behavior of enormous amounts of cells rather than surveying single cells with different phenotypes or genotypes. Here, we report our study of beta-lactamase bacterial drug resistance in a well-designed microfluidic device, which allows us to conduct more controllable experiments, such as controlling the nutrient concentration, switching the culture media, performing parallel experiments, observing single cells, and acquiring time-lapse images. By using GFP as a beta-lactamase indicator and acquiring time-lapse images at the single-cell level, we observed correlations between the bacterial heterogeneous phenotypes and their behavior in different culture media. The feedback loop between the growth rate and the beta-lactamase production suggests that the beta-lactamase bacteria are more resistant in a rich medium than in a relatively poor medium. In the poorest medium, the proportion of dormant cells may increase, which causes a lower death rate in the same generation. Our work may contribute to assaying the antibiotic resistance of pathogenic bacteria in heterogeneous complex media.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / drug effects
  • Bacteria / growth & development
  • Bacteria / metabolism*
  • Culture Media / pharmacology*
  • Drug Resistance, Bacterial / drug effects*
  • Fluorescence
  • Image Processing, Computer-Assisted
  • Microbial Viability / drug effects
  • Phenotype
  • Single-Cell Analysis*
  • beta-Lactamases / metabolism*

Substances

  • Culture Media
  • beta-Lactamases

Grants and funding

This work is partially supported by the National Natural Science Foundation of China (10721403, 11074009, 11174012, 11434001), the Ministry of Science and technology of the People's Republic of China (2009CB918500, 2012AA02A702), the National Fund for Fostering Talents of Basic Science (J0630311). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.