Increased tolerance to commonly used antibiotics in aPseudomonas aeruginosa ex vivoporcine keratitis model

Abstract

AbstractAntibiotics in development are usually tested on rapidly dividing cells in a culture medium and do not reflect the complexity of infectionsin vivo, while testingin vivois limited, expensive and ethically concerning. This often results in the development and subsequent prescription of antibiotics only targeting infections in which pathogens are undergoing rapid cell division and in case of persistent infections like keratitis leads to poor clinical outcomes such as impaired vision or loss of an eye. In this study, we demonstrate antibiotic tolerance ofPseudomonas aeruginosastrains PA01 and PA14 using theex vivoporcine keratitis model in which bacterial physiology more closely mimics infectionsin vivothan in a culture medium.MBEC and MIC were used as a guideline to establish the concentration of applied antibiotics on tissue. Infectedex vivoporcine corneas were treated with therapeutically relevant concentrations of gentamicin, ciprofloxacin and chloramphenicol. Ciprofloxacin was the most potent across all tests demonstrating a positive correlation with MIC but not MBEC. Nonetheless, the results demonstrated that MIC and MBEC concentrations were not sufficient to clear infection even after 18 hours of continuous exposure to the tested antibiotics reflecting the need for novel antibiotics that can target the persistent subpopulation of these pathogens and the ability of theex vivokeratitis model to be a relevant platform to identify novel antibiotics with suitable activities. There was a clear visual distinction between corneas infected with cytotoxic strain PA14 and invasive strain PA01. In this study, both strains PA14 and PA01 showed a high level of antibiotic tolerance, which suggests that in clinical settings the treatment approach could be similar regardless of the causative strain.Data summaryThe authors confirm all supporting data and protocols have been provided within the article or through supplementary data files.