A unique Rickettsia species (Rickettsia thailandii sp. nov.) was identified in an Ixodes granulatus by use of polymerase chain reaction heteroduplex mobility assay by use of segments of the citrate synthase gene. This tick was collected from Rattus rattus from Nakhon Ratchasima province in 1970. Another I. granulatus was infected with Thai tick typhus strain TT-118, Rickettsia honei sp. nov. Stenos, Roux, Walker & Raoult; this tick was removed from a R. rattus collected 4 years later from the same province. Ixodes granulatus is the first tick species in Australasia shown to be infected with R. honei and the unique Rickettsia species.