Introduction. In Brazil, capybaras (Hydrochoerus hydrochaeris) are important hosts for Amblyomma ticks, which in turn can transmit rickettsiae to humans and animals. Therefore, capybaras are potential sentinels for rickettsial infection.Objective. The present study evaluated rickettsial infection in capybaras in different areas of the state of São Paulo, where rickettsiosis has never been reported. Materials and methods. Blood sera from 73 capybaras from six localities in São Paulo were tested by indirect immunofluorescence assay using Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia bellii antigens. Capybara spleens were tested by PCR, targeting a fragment of the rickettsial gltA gene. Ticks were collected from each capybara sample and taxonomically identified to species. Results. A total of 94 positively reacting capybara samples, 19 (26.0%), 25 (34.2%), and 50 (68.5%) capybara sera reacted to R. rickettsii, R. parkeri, and R. bellii, respectively. Twenty-five capybara sera showed titers to R. bellii at least four-fold higher than to any of the other two antigens. These sera were considered homologous to R. bellii. Using the same criteria, 3 capybara sera were considered homologous to R. parkeri. No sera were be considered homologous to R. rickettsii. No rickettsial DNA was detected in capybara spleen samples. Ticks collected on capybaras were Amblyomma dubitatum and Amblyomma cajennense. Conclusions. The first evidence is reported of R. bellii natural infection in vertebrate hosts, and the first evidence of R. parkeri infection in capybaras. While R. parkeri is known to infect and cause disease in humans, no similar evidence for human infection has been indicated by R. bellii.
Lyme disease is caused by bacteria belonging to the complex Borrelia burgdorferi sensu lato, which are transmitted by ticks of the Ixodes ricinus complex. The distribution of these bacteria are restricted to the northern hemisphere (North America, Europe, and Asia). Lyme disease-like cases have been reported in Brazil, but it is possible that another Borrelia species is involved in these cases, and ticks of the genus Amblyomma have been implicated as vectors. Due to these reasons, the disease in Brazil has been referred as Lyme Disease-Simile (LDS). Borrelia anserina, the agent of avian spirochetosis, has a wordwide distribution, where it is transmitted primarily by ticks of the genus Argas. The present study was divided in two chapters: the first one evaluated the presence of Borrelia spp in areas of the state of São Paulo where LDS have been reported. The second chapter reports the first in vitro isolation in BSK medium and molecular characterization of a spirochete strain from Brazil, presumably identified as B. anserina. For the first chapter, a total of 349 adult ticks (Amblyomma cajennense) collected in areas where LDS cases have been reported were processed. In addition, nine human blood or tissue samples from patients with clinical and serological diagnostic of LDS, two samples from BSK medium previously inoculated with samples of skin or blood of LDS patients, and three samples of BSK medium previously inoculated with tick samples were also processed. All these samples were processed for DNA extraction and then tested by nested-PCR employing primers targeting a portion of the flagelin B gene (flaB), which amplify a flaB fragment in all known Borrelia species. All samples were negative by this nested-PCR, showing no evidence of Borrelia sp in the tested samples. The second chapter evaluated an avian spirochete strain originated from Argas miniatus ticks from Pedro Leopoldo municipality, state of Minas Gerais. DNA fragments of the rrs (16S rRNA) and flab genes were amplified by PCR and sequenced to determine phylogenetic similarities. The resulting sequences were 99.8% (483 of 484) and 98.7% (754 of 764) similar to GenBank corresponding sequences of B. anserina rrs and flaB genes, respectively. By neighbor-joining phylogenetic analysis, the flaB sequence of the Brazilian strain clustered in a monophyletic group with the sequence of B. anserina under 100% bootstrap support. The isolate was successfully isolated in BSK medium, with seven passages performed. The spirochete crude antigen, fixed in glass slides, showed strong immunfluorescence reactivity with sera from chickens previously inoculated with the isolate. The spirochete strain isolated in the present study was genetically identified as B. anserina, labeled as strain PL.
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