The importance of vector-borne pathogens has increased in recent years and molecular studies have revealed the presence of these pathogens in new hosts in different parts of the world. Türkiye is one of the important countries where vector-borne pathogens are reported in different hosts. Although there are many studies investigating vector-borne pathogens in cattle, sheep, and goats, there is paucity of data on the presence and distribution of these pathogens in water buffaloes. In this study, it was aimed to investigate Rickettsia species in buffaloes by molecular methods for the first time in Türkiye. In this study, 364 water buffalo blood samples obtained from seven distinct in Sivas province were researched for Rickettsia species using genus-specific PCR assay. Moreover, the partial parts of the ompA and ompB genes of the positive sample were sequenced for species identification and to perform phylogenetic analyses. DNA sequence analysis results showed that the sample identified in the study was infected with Rickettsia aeschlimannii. The obtained DNA sequence belonging to ompA and ompB genes were submitted to GenBank and accession numbers were for ompA gene PQ197208 and ompB gene PQ197209. The BLASTn analyses of both genes revealed that high nucleotide similarities (98.93-100% for ompA and 98.39-100% for ompB) were present between our Ri. aeschlimannii isolate and Ri. aeschlimannii isolates present in the GenBank. Rickettsia aeschlimannii is one of the most important vector-borne pathogens causing infection in humans. This is the first molecular report of Ri. aeschlimannii in water buffalo in Türkiye and the world. This result indicated that the agent circulates in buffaloes. However, large-scale molecular studies are still needed to better understand the contribution of buffaloes to the epidemiology of Ri. aeschlimannii.