Quantitative determination of bacterial replication in vivo

Abstract

A new methodology which permits the quantitative measurement of absolute bacterial replication in vivo is proposed. Mice were inoculated with mixtures of temperature-sensitive mutants and parental wild types, and the changes in the ratios of the two strains were measured. The number of wild-type generations was calculated from the declining ratios over time with the formula n = log [r(0)/r(t)]log2; n is the number of generations, and r(0) and r(t) are the ratio of temperature-sensitive mutants to the parental wild type at time zero and at the times sampled throughout the experiment. The replication rate was determined by regression analysis. A mathematical argument for the formula is presented. Using this technique, we determined the mean generation times of Escherichia coli (33 min) and Pseudomonas aeruginosa (20 min) in the peritoneal cavities of mice, in the face of host clearance mechanisms during the first stages of infection.