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Reding Roman, R. C.

Publications and source records attributed to Reding Roman, R. C..

2 recordsLinked to original sources

Metabolic trade-offs hide unforeseen benefits of plasmids carriage

The link between fitness and reproduction rate is a central tenet in microbiology, and indeed evolutionary biology: Mutants reproducing faster than the dominant wild-type are favoured by selection, but otherwise the mutation is lost. This link was given by Ronald Fisher in 1930 under the assumption that fitness can only change through mutations that boost or hinder growth rate, whence the use of logarithms on growth data by experimentalists. Here I show that logarithms are highly sensitive to sampling times, resulting in fitness estimates that are not constant over the growth of bacterial cultures. This variability invalidates typical selection measurements, and unfit mutants can be co-maintained if they reach their equilibrium. And this is what I observed in competition assays between two Escherichia coli constructs, one of which harbours a non-transmissible plasmid that protects against tetracycline (pGW155B), without using the antibiotic. Despite growing 40% slower than its drug-sensitive counterpart, the construct harbouring the plasmid persisted throughout the competition. And, perhaps more importantly, maintained the plasmid. My study suggests that reliance on growth rate masks that selection on plasmid carriage may be stronger than previously thought--explaining the seemingly-paradoxical abundance of plasmids in nature.

evolutionary biology

Fluorescence photography of patterns and waves of bacterial adaptation at high antibiotic doses

Fisher suggested advantageous genes would spread through populations as a wave so we sought genetic waves in evolving populations, as follows. By fusing a fluorescent marker to a drug efflux protein (AcrB) whose expression provides Escherichia coli with resistance to some antibiotics, we quantified the evolution and spread of drug-resistant E. coli through spacetime using image analysis and quantitative PCR. As is done in hospitals routinely, we exposed the bacterium to a gradient of antibiotic in a disk diffusion drug susceptibility test that we videoed. The videos show complex spatio-genomic patterns redolent of, yet more complex than, Fishers predictions whereby a decelerating wave front of advantageous genes colonises towards the antibiotic source, forming bullseye patterns en route and leaving a wave back of bacterial sub-populations expressing AcrB at decreasing levels away from the drug source. qPCR data show that E. coli sited at rapidly-adapting spatial hotspots gain 2 additional copies of acr, the operon that encodes AcrB, within 24h and imaging data show resistant sub-populations thrive most near the antibiotic source due to non-monotone relationships between inhibition due to antibiotic and distance from the source. In the spirit of Fisher, we provide an explicitly spatial nonlinear diffusion equation that exhibits these properties too. Finally, linear diffusion theory quantifies how the spatial extent of bacterial killing scales with increases in antibiotic dosage, predicting that microbes can survive chemotherapies that have been escalated to 250x the clinical dosage if the antibiotic is diffusion-limited.

evolutionary biology