Data and code from: Antibiotic susceptibility of Escherichia coli is affected by evolutionary history but not by history of elemental limitation
Data files
Mar 10, 2026 version files 37.19 KB
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Figure_2_C-limited_Fitness.csv
5.31 KB
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Figure_2_N-limited_Fitness.csv
5.04 KB
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Figure_3_MIC_analysis_with_ARTool.R
2.24 KB
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Figure_3_N-lim_Expt_MIC_Averaged_Data.csv
4.20 KB
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Figure_3_N-lim_Expt_MIC_Raw_Data.csv
9.31 KB
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Figure_4_A-2_Analysis_with_ARTool.R
918 B
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Figure_4_Ara-2_MIC_Averaged_Data.csv
278 B
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Figure_4_Ara-2_MIC_Raw_Data.csv
2.65 KB
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Figure_S1_N_limitation_of_population_size_.csv
377 B
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README.md
6.88 KB
Abstract
Antibiotic resistance in bacteria is a global public health threat. To understand how the evolution of antibiotic susceptibility is affected by environmental conditions and prior evolutionary history, we worked with populations from the Long-Term Evolution Experiment with Escherichia coli. These populations previously evolved independently for 50,000 generations in an environment without antibiotics, making them an ideal system for studying the effect of evolutionary history on adaptation to new selective pressures. We further evolved five of the LTEE populations, as well as their shared ancestor, under either carbon-limited or nitrogen-limited conditions and then tested intrinsic resistance to four antibiotics. Evolution under elemental limitation did not have a significant impact on resistance to any of the tested antibiotics. However, some LTEE populations did have higher resistance than other populations. Susceptibility also varied within one population, which had the lowest level of resistance to all four antibiotics. We hypothesized that resistance levels might differ between two clades of bacteria that have coexisted within this population for more than 40,000 generations. Interestingly, although antibiotic susceptibility varied within the population, there was not a consistent difference between clades. Instead, one particular clone isolated from the population exhibited higher resistance than the other clones sampled. These findings indicate that antibiotic resistance can vary both within and between experimentally evolved populations, even in the absence of direct selection on resistance. Our results also show that measured levels of susceptibility may depend on stochastic sampling effects during isolation of clones.
Dataset DOI: 10.5061/dryad.1zcrjdg68
Description of the data and file structure
Data includes fitness, minimum inhibitory concentration, and population size measurements for E. coli bacteria experimentally evolved under carbon or nitrogen limitation in the absence of antibiotics.
Files and variables
File: Figure_2_C-limited_Fitness.csv
Description: Data from fitness competitions between ancestral or evolved bacteria and the common competitor REL607. All competitions were conducted in carbon-limited media.
Variables
- LTEE population derived from: Name of Long-Term Evolution Experiment population from which the evolved strain was derived
- Evolved strain ID: Freezer ID number for the evolved competitor
- Ancestor ID: Freezer ID number for the ancestral competitor
- Replicate: Replicate number
- D0- Ancestor colonies: Number of colonies on tetrazolium arabinose (TA) plate on day 0 of competition
- D0- REL607 colonies vs ancestor: Number of colonies on TA plate on day 0 of competition
- D1- Ancestor colonies: Number of colonies on TA plate on day 1 of competition
- D1- REL607 colonies vs ancestor: Number of colonies on TA plate on day 1 of competition
- Generations- Ancestor: Number of generations undergone by the ancestral strain during the competition. Equal to log2("D1- Ancestor colonies"*100/"D0- Ancestor colonies")
- Generations-REL607 vs ancestor: Number of generations undergone by REL607 during competition with the ancestral strain. Equal to log2("D1- REL607 colonies vs ancestor"*100/"D0- REL607 colonies vs ancestor")
- Fitness ancestor (w): Equal to "Generations- Ancestor"/"Generations-REL607 vs ancestor". Unitless
- D0- Evolved colonies: Number of colonies on TA plate on day 0 of competition
- D0- REL607 colonies vs evolved: Number of colonies on TA plate on day 0 of competition
- D1-Evolved colonies: Number of colonies on the TA plate on day 1 of competition
- D1- REL607 colonies vs evolved: Number of colonies on TA plate on day 0 of competition
- Generations- Evolved: Number of generations undergone by the evolved strain during the competition. Equal to log2("D1- Evolved colonies"*100/"D0- Ancestor colonies")
- Generations- REL607 vs evolved: Number of generations undergone by REL607 during competition with the evolved strain. Equal to log2("D1- REL607 colonies vs ancestor"*100/"D0- REL607 colonies vs ancestor")
- Fitness evolved (w): Equal to "Generations- Evolved"/"Generations-REL607 vs evolved". Unitless
- Change in fitness: "Fitness evolved (w)"-"Fitness ancestor (w)".
File: Figure_2_N-limited_Fitness.csv
Description: Data from fitness competitions between ancestral or evolved bacteria and the common competitor REL607. All competitions were conducted in nitrogen-limited media.
Variables
- Same as for Figure_2_C-limited_Fitness.csv above
File: Figure_3_N-lim_Expt_MIC_Averaged_Data.csv
Description: Minimum inhibitory concentrations of ampicillin, tetracycline, tobramycin, and erythromycin for E. coli bacteria evolved under carbon limitation or nitrogen limitation. Values are averages of three replicate measurements.
Variables
- Antibiotic: Name of antibiotic tested
- Strain: CBTn freezer number of tested clone
- Limitation: Identifies the limiting element under which the strain evolved
- Ancestor: Identifies the ancestor of the tested clone
- MIC: Minimum inhibitory concentration (µg/mL). Mean of three replicate measurements.
File: Figure_3_MIC_analysis_with_ARTool.R
Description: R code for statistical analysis of MIC data for E. coli clones evolved under carbon or nitrogen limitation.
File: Figure_4_Ara-2_MIC_Averaged_Data.csv
Description: Minimum inhibitory concentrations of ampicillin, tetracycline, tobramycin, and erythromycin for E. coli clones isolated from population Ara-2 of the Long-Term Evolution Experiment. Values are averages of three replicate MIC measurements.
Variables
- Strain: Freezer ID of tested clone
- Size: Identifies whether the tested clone forms large or small colonies on agar plates
- AMP MIC: Minimum inhibitory concentration (µg/mL) for ampicillin. Mean of three replicate measurements.
- ERY MIC: Minimum inhibitory concentration (µg/mL) for erythromycin. Mean of three replicate measurements.
- TET MIC: Minimum inhibitory concentration (µg/mL) for tetracycline. Mean of three replicate measurements.
- TOB MIC: Minimum inhibitory concentration (µg/mL) for tobramycin. Mean of three replicate measurements.
File: Figure_S1_N_limitation_of_population_size_.csv
Description: Population size of E. coli clones REL606/607 under varying nitrogen concentrations.
Variables
- Ammonium sulfate (mg/L): Concentration of ammonium sulfate in the media (mg/L)
- Colonies: Number of colonies formed on plate after 24 h incubation
- dilution to plate: fold-dilution from culture flask to plate
- cfu/mL: Colony forming units per mL of culture after 24 h incubation. Calculated as "Colonies"*"dilution to plate".
File: Figure_3_N-lim_Expt_MIC_Raw_Data.csv
Description: Minimum inhibitory concentrations of ampicillin, tetracycline, tobramycin, and erythromycin for E. coli bacteria evolved under carbon limitation or nitrogen limitation. Values are individual measurements underlying the average values in the file Figure_3_N-lim_Expt_MIC_Averaged_Data.csv.
Variables
- Antibiotic: Name of antibiotic tested
- Strain: CBTn freezer number of tested clone
- Limitation: Identifies the limiting element under which the strain evolved
- Ancestor: Identifies the ancestor of the tested clone
- MIC (ug/mL): Minimum inhibitory concentration (µg/mL) for the tested antibiotic. Value for a single replicate.
File: Figure_4_A-2_Analysis_with_ARTool.R
Description: R code for statistical analysis of MIC data for E. coli clones isolated from population Ara-2 of the Long-Term Evolution Experiment.
File: Figure_4_Ara-2_MIC_Raw_Data.csv
Description: Minimum inhibitory concentrations of ampicillin, tetracycline, tobramycin, and erythromycin for E. coli clones isolated from population Ara-2 of the Long-Term Evolution Experiment. Values are individual measurements underlying the average values in the file Figure_4_Ara-2_MIC_Averaged_Data.csv.
Variables
- Antibiotic: Name of the antibiotic tested
- Strain: Freezer ID of tested clone
- Size: Identifies whether the tested clone forms large or small colonies on agar plates
- MIC: Minimum inhibitory concentration (µg/mL) for the tested antibiotic. Value for a single replicate.
Code/software
R v4.4.2 with the ARTool and rcompanion packages
