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MOURA, N. O. Detection and molecular characterization of Rickettsia in potential vectors from active focuses of spotted fever in the State of Rio de Janeiro. 2011. 78p. Master’s thesis (Parasitology) – Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, 2012. The Brazilian Spotted Fever (BSF) caused by Spotted Fever Group rickettsiae (SFGR) transmitted by ticks, occurs in Brazil, particularly in the southeast, where deaths are registered. Research indicates that at least three pathogenic species, transmitted by fleas or ticks, circulate in ecotopes of the states of São Paulo and Minas Gerais. In the State of Rio de Janeiro, the lethality of the BSF is high, but only recently epidemiological investigations began in some locations, indicating the participation of new species of ectoparasites in rickettsiae circulation. The objective of this project is to evaluate rickettsiae in ectoparasites collected in areas of suspected, compatible or confirmed BSF human cases in other different cities in the state in recently reported outbreaks. Specifically, the objectives are: a) detection of Rickettsia in possible vectors, ticks and fleas, collected on domestic animals, domesticated and also free environment; b) analysis of Minimum Frequency of Infected Vectors (MFIV) and c) identification, by molecular methods, Rickettsia species detected. Samples were collected between 2008-2009 in locations that lack of information on the maintenance of the transmission cycle of rickettsiae: Noroeste Fluminense (Porciúncula and Itaperuna), Serrana (Trajano de Moraes and São José do Vale do Rio Preto) and Médio Paraiba (Barra do Piraí, Piraí and Volta Redonda). The 2076 identified ectoparasites were collected and processed individually or in groups according to sex, stage of development and degree of engorgement, totalizing 301 samples. The genomic DNA of all samples was extracted and subjected to the search for genes of organisms of the genus Rickettsia (gltA) and SFGR (ompA) by polymerase chain reaction (PCR). We obtained 16 positive samples of the seven cities surveyed. Of these, 15 amplified ompA gene fragment, indicating broad geographic distribution of SFGR in the state. The MFIV was calculated for each species investigated and focus. As additional methodology for detection of rickettsiae in potential vectors, we performed quantitative PCR assays in 231 samples totalizing 1483 analyzed arthropods. Of these, 144 samples amplified gltA gene for Rickettsia sp. The conventional PCR amplicons were cloned and sequences were analyzed. Sequences with 99 and 100% identity with Rickettsia rickettsii, which causes the BSF were diagnosed in Amblyomma cajennense, Amblyomma dubitatum, Boophilus microplus, Ctenocephalides canis and Ctenocephalides felis, and for the first time in the cities of São José do Vale do Rio Preto, Trajano de Moraes, Volta Redonda, Itaperuna and Porciúncula. In A. dubitatum was first recorded presence of sequences with 99% identity with Rickettsia bellii to the state of Rio de Janeiro, and 100% identity with R. rickettsii to Brazil. Similarly, in C. canis sample was obtained sequence with 99% identity with Rickettsia felis in the state of Rio de Janeiro, and 99% identity with R. rickettsii in Brazil. This work reveals the wide geographical distribution of SFGR, including R. rickettsii causes BSF in Rio de Janeiro’ State.

Detecção e caracterização molecular de riquétsias em potenciais vetores procedentes de focos ativos de febre maculosa do Estado do Rio de Janeiro.