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Background  Spotted Fever Group (SFG) Rickettsiae can cause febrile diseases with or without rash in humans worldwide. In Germany only limited data are available about their medical significance. Serological screening tests for antibodies against rickettsiae usually only distinguish between SFG and Typhus Group (TG) Rickettsiae due to the strong cross reactivities within the groups. Seroprevalence rates in dogs, as possible sentinels for tick-borne diseases, could serve as an indicator for the distribution of different  Rickettsia  species.    Methods  In this study, a micro-immunofluorescence assay (micro-IFA) was established for detection and differentiation of antibodies against five  Rickettsia  species in dogs ( R. helvetica ,  R. raoultii ,  R. slovaca ,  R. monacensis  and  R. felis ). Dogs that never left Germany (n = 605) previously investigated with an SFG-ELISA were included in this study and screened at a 1:128 dilution. Endpoint titres of fifty randomly selected seropositive samples of each of the five investigated regions in Germany were determined in order to allow a differentiation of the causative  Rickettsia  species. Sensitivity and specificity of the micro-IFA were compared with ELISA results of the previous study.    Results  A total of 93.9% of the dogs were positive for antibodies of the SFG Rickettsiae at the screening titer of 1:128. Differentiation of SFG Rickettsiae with the micro-IFA was possible in 70.4%, but in 29.6% of the cases the detected antibodies were not differentiable. Considering a clear differentiation by a twofold titre difference between observed reactions, the seroprevalence rates were 66.0% for  R. helvetica,  2.8% for  R. raoultii,  1.6% for  R. slovaca , but no serological reaction could be clearly attributed to  R. monacensis  or  R. felis . No statistically significant regional differences were found for  R. helvetica ,  R. slovaca  and  R. raoultii  comparing the five regions of Germany. Comparison of micro-IFA with ELISA revealed a sensitivity of 82.0% and a specificity of 83.8% for the  Rickettsia  SFG ELISA.    Conclusions  The micro-IFA is a useful serological tool to differentiate antibodies against different  Rickettsia  species in dogs. Seroprevalence rates in dogs correspond to the prevalence rates and distribution of  Rickettsia -carrying tick species.

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Serological differentiation of antibodies against Rickettsia helvetica, R. raoultii, R. slovaca, R. monacensis and R. felis in dogs from Germany by a micro-immunofluorescent antibody test