Suppression of bacterial cell death underlies the antagonistic interaction between ciprofloxacin and tetracycline

Abstract Antibiotic combinations aim to maximise drug treatment efficiency and minimise resistance evolution, but a full understanding of their effect on bacterial cells is lacking. The interaction between the DNA-damaging antibiotic ciprofloxacin and the translation inhibitor tetracycline is antagonistic, resulting in a weaker effect on bacterial growth than expected from each drug individually. While this antagonism has been analysed at the population level, it has not been investigated at the single-cell level. We used a microfluidic device to quantify the antagonism between ciprofloxacin and tetracycline in single bacterial cells under three nutrient conditions. Improved growth results from increased survival of cells under the drug combination compared to ciprofloxacin alone. This effect depends on the initial drug-free growth rate, with better suppression in nutrient-rich conditions. Quantifying the DNA damage response (SOS response) revealed two sub-populations among cells that died upon ciprofloxacin treatment. The larger low-SOS sub-population, which showed increased survival compared to high-SOS cells, explains the stronger antagonistic effect in nutrient-rich conditions. Our results underscore the importance of single-cell quantification in understanding bacterial responses to antibiotic combinations.