| Summary: | Tick-borne agents constitute a growing concern for human and animal health worldwide. <i>Hyalomma aegyptium</i> is a hard tick with a three-host life cycle, whose main hosts for adults are Palearctic tortoises of genus <i>Testudo</i>. Nevertheless, immature ticks can feed on a variety of hosts, representing an important eco-epidemiological issue regarding <i>H. aegyptium</i> pathogens circulation. <i>Hyalomma aegyptium</i> ticks are vectors and/or reservoirs of various pathogenic agents, such as <i>Ehrlichia</i>, <i>Anaplasma</i>, <i>Babesia</i> and <i>Hepatozoon/Hemolivia</i>. <i>Ehrlichia</i> and <i>Anaplasma</i> are emergent tick-borne bacteria with a worldwide distribution and zoonotic potential, responsible for diseases that cause clinical manifestations that grade from acute febrile illness to a fulminant disease characterized by multi-organ system failure, depending on the species. <i>Babesia</i> and <i>Hepatozoon/Hemolivia</i> are tick-borne parasites with increasing importance in multiple species. <i>Testudo graeca</i> tortoises acquired in a large animal market in Doha, Qatar, were screened for a panel of tick-borne pathogens by conventional PCR followed by bidirectional sequencing. The most prevalent agent identified in ticks was <i>Hemolivia mauritanica</i> (28.6%), followed by <i>Candidatus</i> Midichloria mitochondrii (9.5%) and <i>Ehrlichia</i> spp. (4.7%). All samples were negative for <i>Babesia</i> spp. and <i>Hepatozoon</i> spp. Overall, 43% of the examined adult ticks were infected with at least one agent. Only 4.7% of the ticks appeared to be simultaneously infected with two agents, i.e., <i>Ehrlichia</i> spp. and <i>H. mauritanica</i>. This is the first detection of <i>H. mauritanica</i>, <i>Ehrlichia</i> spp. and <i>Candidatus</i> M. mitochondrii in <i>H. aegyptium</i> ticks collected from pet spur-thighed tortoises, in Qatar, a fact which adds to the geographical extension of these agents. The international trade of <i>Testudo</i> tortoises carrying ticks infected with pathogens of veterinary and medical importance deserves strict control, in order to reduce potential exotic diseases.
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