During the two World Wars, Bartonella quintana was responsible for trench fever and is now recognised as an agent of re-emerging infection. Many reports have indicated widespread B . quintana exposure since the 1990s. In order to evaluate its prevalence in ancient populations, we used real-time PCR to detect B . quintana DNA in 400 teeth collected from 145 individuals dating from the 1 st to 19 th centuries in nine archaeological sites, with the presence of negative controls. Fisher’s exact test was used to compare the prevalence of B . quintana in civil and military populations. B . quintana DNA was confirmed in a total of 28/145 (19.3%) individuals, comprising 78 citizens and 67 soldiers, 20.1% and 17.9% of which were positive for B . quintana bacteraemia, respectively. This study analysed previous studies on these ancient samples and showed that the presence of B . quintana infection followed the course of time in human history; a total of 14/15 sites from five European countries had a positive prevalence. The positive rate in soldiers was higher than those of civilians, with 20% and 18.8%, respectively, in the 18 th and 19 th centuries, but the difference in frequency was not significant. These results confirmed the role of dental pulp in diagnosing B . quintana bacteraemia in ancient populations and showed the incidence of B . quintana in both civilians and soldiers.