ABSTRACTCorsica is a touristic mountainous French island in the north-west of the Mediterranean Sea presenting a large diversity of natural environments where many interactions between humans, domestic animals and wild fauna occur. Despite this favourable context, tick-borne pathogens (TBPs) have not systematically been investigated. In this study, a large number of TBPs were screened in ticks collected during one year from domestic and wild hosts in Corsica. More than 1,500 ticks belonging to nine species and five genera (Rhipicephalus,Hyalomma,Dermacentor,IxodesandHaemaphysalis) were analysed individually or pooled (by species, gender, host and locality). A real-time microfluidic PCR was used for high-throughput screening of TBPs DNA. This advanced methodology permitted the simultaneous detection of 29 bacterial and 12 parasitic species (includingBorrelia,Anaplasma,Ehrlichia,Rickettsia,Bartonella,CandidatusNeoehrlichia,Coxiella,Francisella,BabesiaandTheileria). CCHF virus was investigated individually in tick species known to be vectors or carriers of this virus. In almost half of the tick pools (48%), DNA from at least one pathogen was detected and eleven species of TBPs from six genera were reported. TBPs were found in ticks from all collected hosts and were present in more than 80% of the investigated area. The detection of some pathogens DNA confirmed their previous identification in Corsica, such asRickettsia aeschlimannii(23% of pools),Rickettsia slovaca(5%),Anaplasma marginale(4%) andTheileria equi(0.4%), but most TBPs DNA was not reported before in Corsican ticks. This includedAnaplasma phagocytophilum(16%),Rickettsia helvetica(1%), Borrelia afzelii(0.7%), Borrelia miyamotoi(1%), Bartonella henselae(2%),Babesia bigemina(2%) andBabesia ovis(0.5%). The important tick infection rate and the diversity of TBPs reported in this study highlight the probable role of animal reservoir hosts for zoonotic pathogens and human exposure to TBPs on Corsica.