Comparative Activity of Ceftriaxone, Ciprofloxacin, and Gentamicin as a Function of Bacterial Growth Rate Probed by Escherichia coli Chromosome Replication in the Mouse Peritonitis Model

Abstract

Commonly used antibiotics exert their effects predominantly on rapidly growing bacterial cells; yet, the growth dynamics taking place during infection in a complex host environment remain largely unknown. Hence, a means to measure <i>in situ</i> bacterial growth rate is essential to predict the outcome of antibacterial treatment. We have recently validated chromosome replication as a readout of <i>in situ</i> bacterial growth rate during <i>Escherichia coli</i> infection in the mouse peritonitis model. By the use of two complementary methods (quantitative PCR and fluorescence microscopy) for differential genome origin and terminus copy number quantification, we demonstrated the ability to track bacterial growth rate, both on a population average level and on a single-cell level, from one single biological specimen. Here, we asked whether the <i>in situ</i> growth rate predicts antibiotic treatment effect during infection in the same model. Parallel <i>in vitro</i> growth experiments were conducted as a proof of concept. Our data demonstrate that the activities of the commonly used antibiotics ceftriaxone and gentamicin correlated with pretreatment bacterial growth rate; both drugs performed better during rapid growth than during slow growth. Conversely, ciprofloxacin was less sensitive to bacterial growth rate, both in a homogenous <i>in vitro</i> bacterial population and in a more heterogeneous <i>in vivo</i> bacterial population. The method serves as a platform to test any antibiotic's dependency on active <i>in situ</i> bacterial growth. Improved insight into this relationship <i>in vivo</i> could ultimately prove helpful in evaluating future antibacterial strategies.