Owing to the potential role of the tick Amblyomma cooperi in the enzootic cycle of Rickettsia rickettsii, the etiologic agent of Brazilian spotted fever (BSF), this study evaluated infection by Rickettsia species in A. cooperi ticks collected from an area in Brazil where BSF is endemic. Among a total of 40 A. cooperi adult ticks collected in an area of BSF endemicity in the state of São Paulo, PCR analysis detected DNA of Rickettsia bellii in 16 ticks (40%), and 3 other ticks (7.5%) were positive for a previously unidentified spotted-fever-group (SFG) rickettsia. Cultivation in Vero cell cultures by the shell vial technique with individual A. cooperi ticks resulted in two isolates of R. bellii and one isolate genotypically characterized as an SFG rickettsia. The two R. bellii isolates were established in Vero cell cultures in the laboratory and were confirmed to be R. bellii by molecular analysis of the gltA and 17-kDa protein-encoding genes and by electron microscopic analysis. The SFG rickettsial isolate could not be stably passaged in cell culture in the laboratory, but molecular analysis of early passages suggested that it was closely related to Rickettsia parkeri, Rickettsia africae, and Rickettsia sibirica. These results do not support the role of A. cooperi in the ecology of R. rickettsii in the area studied, but they add two more species of rickettsiae to the poorly developed list of species occurring in ticks in South America.

Spotted-fever-caused Rickettsia rickettsii infection is in Brazil the major tick-borne zoonotic disease. Recently, a second and milder human rickettsiosis caused by an agent genetically related to R. parkeri was discovered in the country (Atlantic rainforest strain). Both diseases clearly have an ecological background linked to a few tick species and their environment. Capybaras (Hydrochoerus hydrochaeris) and Amblyomma cajennense ticks in urban and rural areas close to water sources are the main and long-known epidemiological feature behind R. rickettsii-caused spotted-fever. Unfortunately, this ecological background seems to be increasing in the country and disease spreading may be foreseen. Metropolitan area of São Paulo, the most populous of the country, is embedded in Atlantic rainforest that harbors another important R. rickettsii vector, the tick Amblyomma aureolatum. Thus, at the city–forest interface, dogs carry infected ticks to human dwellings and human infection occurs. A role for R. rickettsii vectoring to humans of a third tick species, Rhipicephalus sanguineus in Brazil, has not been proven; however, there is circumstantial evidence for that. A R. parkeri-like strain was found in A. ovale ticks from Atlantic rainforest and was shown to be responsible for a milder febrile human disease. Rickettsia-infected A. ovale ticks are known to be spread over large areas along the Atlantic coast of the country, and diagnosis of human infection is increasing with awareness and proper diagnostic tools. In this review, ecological features of the tick species mentioned, and that are important for Rickettsia transmission to humans, are updated and discussed. Specific knowledge gaps in the epidemiology of such diseases are highlighted to guide forthcoming research.

This study investigated rickettsial infection in animals, humans, ticks, and fleas collected in five areas of the state of São Paulo. Eight flea species (Adoratopsylla antiquorum antiquorum, Ctenocephalides felis felis, Polygenis atopus, Polygenis rimatus, Polygenis roberti roberti, Polygenis tripus, Rhopalopsyllus lugubris, and Rhopalopsyllus lutzi lutzi), and five tick species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Ixodes loricatus, and Rhipicephalus sanguineus) were collected from dogs, cats, and opossums. Rickettsia felis was the only rickettsia found infecting fleas, whereas Rickettsia bellii was the only agent infecting ticks, but no animal or human blood was shown to contain rickettsial DNA. Testing animal and human sera by indirect immunofluorescence assay against four rickettsia antigens (R. rickettsii, R. parkeri, R. felis, and R. bellii), some opossum, dog, horse, and human sera reacted to R. rickettsii with titers at least four-fold higher than to the other three rickettsial antigens. These sera were considered to have a predominant antibody response to R. rickettsii. Using the same criteria, opossum, dog, and horse sera showed predominant antibody response to R. parkeri or a very closely related genotype. Our serological results suggest that both R. rickettsii and R. parkeri infected animals and/or humans in the studied areas.Key words: Rickettsia -spotted fever -fleas -ticks -opossum -domestic animals Rickettsiae are obligate intracellular bacteria that have been classically divided into two groups: the typhus group (TG), composed of Rickettsia prowazekii and Rickettsia typhi, which are associated with lice and fleas, respectively; and the spotted fever group (SFG), which includes more than 20 valid species, mostly associated with ticks (e.g., Rickettsia rickettsii, Rickettsia parkeri), and at least one species associated with fleas (Rickettsia felis). Other species, such as Rickettsia bellii and Rickettsia canadensis, both associated with ticks, have been considered a distinct group sharing antigenic, genetic and ecological characters with both TG and SFG rickettsiae (Parola et al. 2005b). Since all SFG Rickettsia share common outer membrane antigens, conventional serological assays employing a single Rickettsia antigen (e.g., R. rickettsii) allow detecting a SFG infection, but they do not identify the Rickettsia species responsible for the infection, even in the presence of high titers (La Scola & Raoult 1997). For a more specific serological diagnosis, it is recommended techniques such as testing serum against all known Rickettsia species present in a given area, or serum cross-absorption tests (La Scola & Raoult 1997, Parola et al. 2005b).For many years, SFG rickettsiosis in the Americas was thought to be caused only by R. rickettsii, transmitted by ticks (Weiss & Moulder 1984). This rickettsia has been reported in several countries from North, Central, and South America causing an acute, highly lethal febrile disease receiving different names according to the country...

Brazilian spotted fever (BSF) is a highly lethal disease caused by Rickettsia rickettsii. In the present study, rickettsial infection was evaluated in 669 Amblyomma aureolatum adult ticks collected from naturally infested dogs in Taiaçupeba, a BSF-endemic area in the state of São Paulo. Ten (1.49%) ticks were infected with Rickettsia bellii, and 6 (0.89%) ticks were infected with R. rickettsii. Both Rickettsia species were isolated and established in Vero cell cultures. The Rickettsia isolates were characterized by molecular analyses, sequencing fragments of different rickettsial genes. Our results suggest that A. aureolatum is an important vector of R. rickettsii in Brazil.

The present study evaluated the rickettsial infection among dogs living in the rural and urban areas of Monte Negro, state of Rondônia, western Brazilian Amazon. Canine sera were tested by the indirect immunofluorescence assay (IFA) using six rickettsial antigens: Rickettsia bellii, Rickettsia amblyommii, Rickettsia rhipicephali, Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia felis. While the first three Rickettsia species are known to occur in the study site, the latter three species are known to occur in southeastern Brazil. For each serum, end point titer reacting with each Rickettsia antigen was determined. Serum showing for a Rickettsia species titer at least fourfold higher than that observed for any other Ricketttsia species was considered homologous to the first Rickettsia species or to a very closely related genotype. A total of 164 rural and 153 urban dogs were tested. Overall, 19 (11.6%) and 6 (3.9%) dogs from rural and urban areas, respectively, reacted positively to at least one Rickettsia species. In the rural area, three sera showed titers to R. parkeri at least four-fold higher than any of the other five antigens. These sera were considered to be homologous to R. parkeri or a very closely related genotype. Using the same criteria, two rural sera were considered homologous to R. amblyommii, two other rural sera to R. rhipicephali, and one urban serum to R. parkeri. Because dogs living in the rural area of Monte Negro are commonly infested by the same tick species infesting humans, they indeed serve as sentinels for human rickettsial diseases. Thus, humans living in Monte Negro are likely to be infected by at least three Rickettsia species: R. parkeri, R. amblyommii, and R. rhipicephali. While R. parkeri is a known human pathogen, further studies are required to verify the potential role of R. amblyommii and R. rhipicephali as human pathogens.

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.

The present study reports field data of ticks infesting wild carnivores captured from July 1998 to September 2004 in Brazil. Additional data were obtained from one tick collection and from previous published data of ticks on carnivores in Brazil. During field work, a total of 3437 ticks were collected from 89 Cerdocyon thous (crab-eating fox), 58 Chrysocyon brachyurus (maned wolf), 30 Puma concolor (puma), 26 Panthera onca (jaguar), 12 Procyon cancrivorus (crab-eating raccoon), 4 Speothos venaticus (bush dog), 6 Pseudalopex vetulus (hoary fox), 6 Nasua nasua (coati), 6 Leopardus pardalis (ocelot), 2 Leopardus tigrinus (oncilla), 1 Leopardus wiedii (margay), 1 Herpailurus yagouaroundi (jaguarundi), 1 Oncifelis colocolo (pampas cat), 1 Eira barbara (tayara), 1 Galictis vittata (grison), 1 Lontra longicaudis (neotropical otter), and 1 Potus flavus (kinkajou). Data obtained from the Acari Collection IBSP included a total of 381 tick specimens collected on 13 C. thous, 8 C. brachyurus, 3 P. concolor, 10 P. onca, 3 P. cancrivorus, 4 N. nasua, 1 L. pardalis, 1 L. wiedii, 4 H. yagouaroundi, 1 Galictis cuja (lesser grison), and 1 L. longicaudis. The only tick-infested carnivore species previously reported in Brazil, for which we do not present any field data are Pseudalopex gymnocercus (pampas fox), Conepatus chinga (Molina's hog-nosed skunk), and Conepatus semistriatus (striped hog-nosed skunk). We report the first tick records in Brazil on two Felidae species (O. colocolo, H. yagouaroundi), two Canidae species (P. vetulus, S. venaticus), one Procyonidae species (P. flavus) and one Mustelidae (E. barbara). Tick infestation remains unreported for 5 of the 26 Carnivora species native in Brazil: Oncifelis geoffroyi (Geoffroy's cat), Atelocynus microtis (short-eared dog), Pteronura brasiliensis (giant otter), Mustela africana (Amazon weasel), and Bassaricyon gabbii (olingo). Our field data comprise 16 tick species represented by the genera Amblyomma (12 species), Ixodes (1 species), Dermacentor (1 species), Rhipicephalus (1 species), and Boophilus (1 species). Additional 5 tick species (3 Amblyomma species and 1 species from each of the genera Ixodes and Ornithodoros) were reported in the literature. The most common ticks on Carnivora hosts were Amblyomma ovale (found on 14 host species), Amblyomma cajennense (10 species), Amblyomma aureolatum (10 species), Amblyomma tigrinum (7 species), Amblyomma parvum (7 species), and Boophilus microplus (7 species).

The present study reports a collection of Amblyomma spp. ticks in birds from several areas of the state of São Paulo, Brazil. A total of 568 tick specimens (404 larvae, 164 nymphs) were collected from 261 bird specimens. From these ticks, 204 (36%) specimens (94 larvae, 110 nymphs) were reared to the adult stage, being identified as Amblyomma longirostre (94 larvae, 90 nymphs), Amblyomma calcaratum (13 nymphs), Amblyomma nodosum (2 nymphs), and Amblyomma cajennense (5 nymphs). Additionally, 39 larvae reared to the nymphal stage and 8 nymphs that died before reaching the adult stage were identified as A. longirostre according to peculiar characters inherent to the nymphal stage of this species: scutum elongate, and hypostome pointed. The remaining 271 larvae and 46 nymphs were identified as Amblyomma sp. Ticks were collected from 51 species of birds distributed in 22 bird families and 6 orders. The order Passeriformes constituted the vast majority of the records, comprising 253 (97%) out of the 261 infested birds. Subadults of A. longirostre were identified from 35 species of Passeriformes, comprising 11 families (Cardinalidae, Dendrocolaptidae, Fringillidae, Furnariidae, Parulidae, Pipridae, Thamnophilidae, Thraupidae, Turdidae, Tyrannidae, and Vireonidae), and from 1 species of a non-passerine bird, a puffbird (Bucconidae). Subadults of A. calcaratum were identified from 5 species of Passeriformes, comprising 5 families (Cardinalinae, Conopophagidae, Pipridae, Thamnophilidae and Turdidae). Subadults of A. nodosum were identified from 2 species of Passeriformes, comprising two bird families (Thamnophilidae and Pipridae). Subadults of A. cajennense were identified from 2 species of non-passerine birds, belonging to 2 different orders (Ciconiiformes: Threskiornithidae, and Gruiformes: Cariamidae). Birds were usually infested by few ticks (mean infestation of 2.2 ticks per bird; range: 1-16). Currently, 82 bird species are known to be infested by immature stages of A. longirostre, with the vast majority [74 (90%)] being Passeriformes. Our results showed that Passeriformes seems to be primary hosts for subadult stages of A. longirostre, A. calcaratum, and A. nodosum. However, arboreal passerine birds seem to be the most important hosts for A. longirostre whereas ground-feeding passerine birds seem to be the most important for both A. calcaratum and A. nodosum. In contrast, the parasitism of birds by subadults of A. cajennense has been restricted to non-passerine birds.