Ticks are vectors of many zoonotic pathogens of clinical relevance, including Anaplasma and Rickettsia species. Since few data about these tick-borne pathogens are available in the Canary Islands, the aim of the present study was to screen their presence in questing and feeding ticks on the island of Tenerife. A total of 81 ticks was removed from six hedgehogs, and eight ticks were collected from the grass. PCR assays for tick species identification based on the LSU rRNA gene, and detection of Anaplasma spp. and Rickettsia spp. by targeting the 16s rRNA and gltA gene, respectively, were carried out. Rhipicephalus sanguineus and Haemaphysalis formosensis tick species were identified, with Rickettsia hoogstraalii detected in H. formosensis. In addition, Rickettsia helvetica and one unidentified Rickettsia species were detected. The overall prevalence of Rickettsia in ticks was 12.2%, and none harbored Anaplasma sp. This work constitutes the first identification in the Canary Islands of pathogenic R. helvetica and the species of unknown pathogenicity R. hoogstraalii in questing ticks. The clinical relevance of the pathogens identified highlights the need for studies with increased sample size and locations, including potential hosts, as well as warning the population about the relevance of ticks as vectors.