Host-seeking Ixodes ricinus (L.) (Acari: Ixodidae) ticks were collected systematically, from May to September 2006, at selected sites in southern Germany, including a large city park in Munich. Polymerase chain reactions for amplification of genes of the rickettsial citrate synthase (gltA), the outer membrane proteins A and B (ompA and ompB), and the 16S rDNA were used to investigate 2,861 specimens (adults and nymphs). GltA sequences of spotted fever group rickettsiae were detected in 151 of all samples (5.3%; 95% CI = 4.3-6.2%). Sequencing revealed Rickettsia helvetica in 91.4% of the samples and R. monacensis in 8.6%. Amplification of ompA was not possible for R. helvetica, but in all except one of the R. monacensis. The results were analyzed statistically to test the effects of season, location, developmental stage, and gender of the tick on prevalence of Rickettsia spp. Although rickettsial DNA was detected in all investigated sites, sites in natural forest areas had significantly higher prevalences than sites in landscaped city parks. Adult female and male ticks had a similar prevalence and were significantly more often infected than nymphs. Monthly differences were not statistically significant. These results clearly show that R. helvetica is widespread throughout the study region and could result in a threat to public health in areas of high prevalence.