Amblyomma triguttatum triguttatum is assumed to be endemic to south-western Western Australia (including Barrow Island), Queensland (excluding Cape York Peninsula), and New South Wales, south to Dubbo and Barham. The species has been recorded on a range of mammalian hosts including macropods and domestic animals. In Queensland, A. triguttatum triguttatum is implicated in the epidemiology of Q fever. In 2000, the species was detected on southern Yorke Peninsula in South Australia. We aimed to identify A. triguttatum triguttatum's hosts through trapping, sampling of carcasses, and opportunistic capture of vertebrates on Yorke Peninsula. A. triguttatum triguttatum was removed from black rats (Rattus rattus), wild rabbits (Oryctolagus cuniculus), western grey kangaroos (Macropus fuliginosus), Tammar wallabies (M. eugenii eugenii), domesticated cats and dogs, and humans. Before this study, A. triguttatum triguttatum had not been found on black rats or rabbits in the wild. This research has implications for the management of wildlife, livestock, and visitors on Yorke Peninsula. The potential for A. triguttatum triguttatum to spread to other areas of Yorke Peninsula and South Australia is considerable, as visitors (tourists) to southern Yorke Peninsula report the presence of ticks both on themselves and among camping equipment on arriving home.