[Frontiers in Bioscience 14, 4815-4824, June 1, 2009]

Evolution of altruists and cheaters in near-isogenic populations of Escherichia coli

Denise R. Clark1, Tia M. Alton1, Alexander Bajorek1, Patricia Holden2, Lee A. Dugatkin1, Ronald M. Atlas1, Michael H. Perlin1

1University of Louisville, Louisville, Kentucky 40292 USA, 2 Present Address: Ohio University, Athens, OH 45701 USA

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
4. Results
4.1. Testing membrane selective permeability of competition flasks: whole cells do not cross, while beta-lactamase and ampicillin cross freely
4.2. Growth rates of starting strains are approximately the same, but all plasmid-bearing strains are not "created equal"
4.3. Competitions to determine whether background strains bearing the same plasmid grow equally well in the presence of antibiotic
4.4. Testing beta-lactamase location in liquid cultures of resistant strains
4.5. Competitions for 120 h
4.6. Other kinds of Cheaters
5. Discussion
6. Acknowledgements
7. References

1. ABSTRACT

Emergence of antibiotic-resistant bacteria threatens the continued efficacy of many critical drugs used to treat serious infections. What if such resistant organisms could also act as altruists and "share" their resistance with sensitive cohorts without any actual genetic exchange? We competed resistant strains that differ solely in their ability to secrete a plasmid-encoded beta-lactamase. Sensitive strains were otherwise isogenic with their resistant counterparts and were either plasmid-free or contained a "Dummy" plasmid of roughly the same size as that of the resistance plasmids. Absent antibiotic selection, plasmid-free sensitive strains outperformed the plasmid-bearing strains. In the presence of ampicillin, the outcome depended on whether the resistant strain secreted its beta-lactamase (Altruist) or retained it (Selfish). In the latter case, only resistant cells survived. When beta-lactamase was secreted, some sensitive cohorts were also provided protection, with the largest fitness increase provided to plasmid-free cells. However, some Altruist strains appeared to be at a disadvantage, as a great deal of their enzyme broke off cells. Thus, additional variables must be considered when designing microbial competition experiments.