ABSTRACT. Navarrete M.G., Cordeiro M.D., Silva C.B., Pires, M.S., Ribeiro C.C.D.U., Cabezas-Cruz A., Massard C.L., López E.R & Foseca A.H. Molecular detection of Ehrlichia canis and Babesia canis vogeli in Rhipicephalus sanguineus sensu lato ticks from Cuba. [Detecção molecular de Ehrlichia canis e Babesia canis vogeli em Rhipicephalus sanguineus sensu lato de carrapatos em Cuba.] Revista Brasileira de Medicina Veterinária, 38(supl. 3):63-67, 2016. Programa de Pós-Graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23897-970, Brasil. E-mail: firstname.lastname@example.org Ticks (Acari: Ixodidae) are of relevant medical and veterinary importance worldwide because of the range of pathogens they can transmit. A survey was carried out to identify Babesia spp. and Ehrlichia spp. in ticks obtained from dogs of Cuba. A total of 431 ticks were collected from 378 dogs and the specimens were properly identified according to a taxonomic key for the genera. All ticks were identified as belong to the R. sanguineus sensu lato (s. l.) species. Genomic DNA was extracted from ticks with protocol using phenol/chloroform. Ticks were organized in pools and the DNA extracted were tested by a nested polymerase chain reaction (nPCR) to amplify 398 base pairs (bp) from the 16S ribosomal DNA (rDNA) of E. canis, and to a PCR to amplify approximately 560 bp from 18S ribosomal DNA (rDNA). Of the 49 pools tested, 8.16% (n=4/49) were positive for the E. canis by nPCR targeting the16S rDNA gene and only one pool (n=1/49; 2.04%) was positive for the gene 18S rDNA for Babesia canis. The four sequences obtained for the 16S rDNA fragment were identical to each other and resulted in 100% identity with E. canis from different countries. The obtained sequence of the 18S rDNA gene for Babesia spp. presented similarity of 100% with Babesia canis vogeli when compared to sequences deposited in Genbank. This is the first molecular detection of these agents in the tick R. sanguineus s. l. in Cuba.