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Background: Our studies undertaken between 2006-2015 have shown that the most frequent species of ticks in Romania is Ixodes ricinus. It was found that I. ricinus is the main vector in Romania for the Borrelia burgdorferi s.l. and for the TBEv, but no data were available for Francisella tularensis and CCHFv infectious agents occurrence into I.ricinus and Hyalomma sp. Three Romanian counties were selected as ticks sampling sites (Sibiu, Tulcea and Giurgiu), with this occasion we collected ticks from the vegetation and from the livestock and reptile fauna. Among the reptiles (Tulcea county), Testudo graeca ibera (TGI) is a well represented species. Samples of ectoparasites obtained from TGI and livestock collected during the years: 2006-2007 and 2014 - 2015 (April-June) showed that the majority of ticks are represented by Hyalomma.aegyptium and H. marginatum. Over 400 I. ricinus and Hyalomma sp. ticks were collected and analyzed by Real-Time PCR methods (including the new TicKitqPCR detection concept; project funded by the MEN-UEFISCDIPNII "Partnerships in priority areas" program, National Research Grant No. 295/2014") that give us results on Borrelia burgdorferi, Francisella tularensis, TBEv and CCHFv (BFTC) presence in the vectors. Methods & Materials: Total RNA and DNA were extracted and analized by in house real-time PCR reagents (included in the new TicKitqPCR detection concept) and 2 commercial kits for the BFTC detection in the I. ricinus and Hyalomma sp.pools. Results: Specific DNAs from B. burgdorferi sl. were detected (Fla B gene target) in 20% of I. ricinus ticks and specific DNAs for F. tularensis were detected (IS Ftu2 genomic isertion-like element) in 2% of the same vector species. Specific RNAs from TBEv were detected (3' UTR-genomic region) in < 1% of I.ricinus pools. No specific CCHFv RNAs were detected (S -genomic region) among the Hyalomma sp. pools. Conclusion: The results strengthen the concern that already exists in Romania, for the enhancement the surveillance and the control measures for the tick populations but also for the means of active information of the human population about the danger of the diseases transmitted by ticks in some risk areas. [ABSTRACT FROM AUTHOR]