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SUMMARYThe tick-borne bacteriumRickettsia rickettsiiis the aetiological agent of Brazilian spotted fever (BSF). The present study evaluated tick infestations on wild and domestic animals, and the rickettsial infection in these animals and their ticks in 7 forest areas adjacent to human communities in the São Paulo Metropolitan Area (SPMA). The results were compared to ecological traits of each sampled area. Two main tick species,Amblyomma aureolatumandRhipicephalus sanguineus,were collected from dogs. The major ticks found on small mammals and birds wereIxodes loricatusandAmblyomma longirostre, respectively. Both anti-R. rickettsiiantibodies andR. rickettsii-infected ticks were detected on dogs from only 2 areas in the southern part of the SPMA, which were considered to be endemic for BSF; the remaining 5 areas were considered to be non-endemic. Ecologically, the BSF-endemic areas clearly differed from the non-endemic areas by the presence of significantly more degraded forest patches in the former. The present results corroborate historical observations that have indicated that all human cases of BSF in the SPMA were contracted in the southern part of this metropolitan area. However, not all forest patches in the southern part of the SPMA were shown to be associated with BSF endemism.

Recently, a novel human rickettsiosis, namely Atlantic rainforest spotted fever, was described in Brazil. We herein report results of a survey led around the index case in an Atlantic rainforest reserve in Peruibe municipality, southeastern Brazil. A Rickettsia parkeri-like agent (Rickettsia sp. Atlantic rainforest genotype) and Ricketsia bellii were isolated from adult Amblyomma ovale ticks collected from dogs. Molecular evidence of infection with strain Atlantic rainforest was obtained for 30 (12.9%) of 232 A. ovale adult ticks collected from dogs. As many as 88.6% of the 35 examined dogs had anti-Rickettsia antibodies, with endpoint titres at their highest to R. parkeri. High correlation among antibody titres in dogs, A. ovale infestations, and access to rainforest was observed. Amblyomma ovale subadults were found predominantly on a rodent species (Euryoryzomys russatus). From 17 E. russatus tested, 6 (35.3%) displayed anti-Rickettsia antibodies, with endpoint titres highest to R. parkeri. It is concluded that Atlantic rainforest genotype circulates in this Atlantic rainforest area at relatively high levels. Dogs get infected when bitten by A. ovale ticks in the forest, and carry infected ticks to households. The role of E. russatus as an amplifier host of Rickettsia to A. ovale ticks deserves investigation.

We experimentally infected Amblyomma aureolatum ticks with the bacterium Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (RMSF). These ticks are a vector for RMSF in Brazil. R. rickettsii was efficiently conserved by both transstadial maintenance and vertical (transovarial) transmission to 100% of the ticks through 4 laboratory generations. However, lower reproductive performance and survival of infected females was attributed to R. rickettsii infection. Therefore, because of the high susceptibility of A. aureolatum ticks to R. rickettsii infection, the deleterious effect that the bacterium causes in these ticks may contribute to the low infection rates (<1%) usually reported among field populations of A. aureolatum ticks in RMSF-endemic areas of Brazil. Because the number of infected ticks would gradually decrease after each generation, it seems unlikely that A. aureolatum ticks could sustain R. rickettsii infection over multiple successive generations solely by vertical transmission.

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Ticks collected in the last two decades from free-living and captive wild animals from 28 municipalities of the Brazilian state of Mato Grosso were identified and tested using molecular methods for the presence of rickettsial agents. A total of 4467 ticks (229 larvae, 1676 nymphs, 1565 males, 997 females) representing 27 ixodid species were collected from 235 species of amphibians, reptiles, birds, and mammals from three different ecoregions (Pantanal, Cerrado, and Amazonia). The species Amblyomma parkeri, Amblyomma romitii, Amblyomma varium and Ixodes luciae are reported for the first time in the state of Mato Grosso. Amongst 538 ticks tested by molecular methods for rickettsial infection, we detected 'Candidatus Rickettsia amblyommii' infecting Amblyomma cajennense sensu stricto and Amblyomma coelebs, Rickettsia sp. strain Atlantic rainforest infecting Amblyomma ovale, Rickettsia sp. strain NOD infecting Amblyomma nodosum, and 'Candidatus Rickettsia andeanae' infecting Amblyomma sculptum. Our results represent an impressive expansion of knowledge on tick fauna and rickettsiae and are essential for understanding the ecology of ticks and tick-borne diseases in the Neotropical region, particularly in midwestern Brazil.

The present study aimed to evaluate under controlled conditions the acquisition of Hepatozoon canis by Amblyomma ovale after feeding on infected dogs, and the subsequent induction of infection in uninfected dogs that ingested the experimentally infected ticks. Two H. canis naturally infected dogs were infested with A. ovale adult ticks derived from an uninfected laboratory tick colony. After feeding, two A. ovale females presented H. canis oocysts in the hemolymph at the first and fourth days after removal of ticks from dogs. The oocysts had an average size of 244.34 microm x 255.46 microm. Three uninfected dogs were fed with ticks previously fed on the infected dogs. Only one dog became infected 32 days after oral inoculation, presenting circulating gametocytes, parasitemia less than 1%, and positive PCR confirmed to be H. canis by DNA sequencing. The results obtained indicated A. ovale ticks as potential vector of H. canis in rural areas of Brazil.

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