Lizeth Taylor

First Report of the Isolation and Molecular Characterization of Rickettsia amblyommii and Rickettsia felis in Central America

During 2010, 15 adult ticks, identified as Amblyomma cajennense, were collected from horses in Cahuita and Turrialba districts, whereas 7 fleas, identified as Ctenocephalides felis, were collected from a dog in San Jose city, Costa Rica. In the laboratory, three A. cajennense specimens, two from Cahuita and one from Turrialba, were individually processed for rickettsial isolation in cell culture, as was a pool of seven fleas. Rickettsiae were successfully isolated and established in Vero cell culture from the three ticks and from a pool of seven fleas in C6/36 cell culture. The three tick isolates were genotypically identified as Rickettsia amblyommii, and the flea isolate was identified as Rickettsia felis through DNA sequencing of portions of the rickettsial genes gltA, ompA, and ompB of each isolate. In addition, other seven ticks were shown to contain rickettsial DNA. Polymerase chain reaction products of at least two of these ticks were sequenced and also showed to correspond to R. amblyommii. Overall, 66.7% (10/15) of the A. cajennense adult ticks were found to be infected with rickettsiae. This is the first report of a successful isolation in cell culture of R. amblyommii and R. felis from Central America.

Rickettsia felis in Ctenocephalides felis from Guatemala and Costa Rica

Rickettsia felis is an emerging human pathogen associated primarily with the cat flea Ctenocephalides felis. In this study, we investigated the presence of Rickettsia felis in C. felis from Guatemala and Costa Rica. Ctenocephalides felis were collected directly from dogs and cats, and analyzed by polymerase chain reaction for Rickettsia-specific fragments of 17-kDa protein, OmpA, and citrate synthase genes. Rickettsia DNA was detected in 64% (55 of 86) and 58% (47 of 81) of flea pools in Guatemala and Costa Rica, respectively. Sequencing of gltA fragments identified R. felis genotype URRWXCal(2) in samples from both countries, and genotype Rf2125 in Costa Rica. This is the first report of R. felis in Guatemala and of genotype Rf2125 in Costa Rica. The extensive presence of this pathogen in countries of Central America stresses the need for increased awareness and diagnosis.

Exposure of dogs to spotted fever group rickettsiae in urban sites associated with human rickettsioses in Costa Rica.

The zoonotic transmission cycles of Rickettsia rickettsii and other spotted fever group (SFG) rickettsiae in Latin America have usually been associated with rural or sylvatic environments, although domestic dogs can be implicated in more populated settings. In this study, exposure of dogs to SFG rickettsiae in the Greater Metropolitan Area of Costa Rica was investigated. Dogs from sites associated with human cases and from dog shelters were evaluated by indirect immunofluorescence assay (IFA) using antigen of SFG rickettsiae. Rickettsia spp. were detected in ectoparasites by polymerase chain reaction (PCR). A total 18.5% (31/168) of dogs associated with human cases and 6.8% (11/161) of dogs in shelters had IgG end titers≥64 to Rickettsia spp. The odds of being seropositive were greater in dogs from areas associated with human cases when compared to shelters (OR: 3.2; 95% C.I: 1.5-5.6). Rhipicephalus sanguineus sensu lato (s. l.) was present in all sites associated with human cases. Rickettsia felis URRWXCal2 and R. felis-like RF2125 were detected in Ctenocephalides felis, and Rickettsia sp. IbR/CRC in Ixodes boliviensis. Results demonstrate that dogs from the main urban center of Costa Rica have been exposed to SFG rickettsiae, especially in areas with known human infection. Both human and animal health sectors must be aware of possible rickettsial diseases in urban areas, where dogs may also serve as sentinels for human infection.