Molecular screening of tick-borne microorganisms in ticks from rural areas of Panama, with the first record of Ehrlichia minasensis in Rhipicephalus microplus from Central America.

During a molecular screening of rickettsial, borrelial and protozoal agents in ticks from rural areas of Panama, we detected DNA of Rickettsia amblyommatis in four Amblyomma mixtum collected from horses in a locality in Los Santos province. Ehrlichia minasensis and Anaplasma marginale were identified from three and two Rhipicephalus microplus, respectively, collected on cows from Chiriqui province. No DNA from other tick-borne agents was found. These results increase the information of R. amblyommatis and A. marginale in Panama, and also mark the first record of E. minasensis in Central America.

Molecular screening for tick-borne bacteria and hematozoa in Ixodes cf. boliviensis and Ixodes tapirus (Ixodida: Ixodidae) from western highlands of Panama.

The first molecular screening for Rickettsia, Anaplasma, Ehrlichia, Borrelia, Babesia and Hepatozoon was carried out in questing Ixodes cf. boliviensis and Ixodes tapirus from Talamanca Mountains, Panama, using specific primers, sequencing and phylogeny. Phylogenetic analyses for the microorganisms in Ixodes cf. boliviensis confirmed the presence of Rickettsia sp. strain IbR/CRC endosymbiont (26/27 ticks), three genotypes of the Borrelia burgdorferi (sensu lato) complex (4/27 ticks), Babesia odocoilei (1/27 ticks), and Hepatozoon sp. (2/27 ticks), tentatively designated Hepatozoon sp. strain Chiriquensis. Phylogenetic analyses for the microorganisms in I. tapirus revealed an undescribed Rickettsia sp., tentatively designated Rickettsia sp. strain Itapirus LQ (6/6 ticks), and Anaplasma phagocytophilum (2/6 ticks). To the best of our knowledge, this is the first report of B. burgdorferi (s.l.) complex, A. phagocytophilum, B. odocoilei, and Hepatozoon sp. in Ixodes ticks from Central America, and also the first detection of Rickettsia spp. in Ixodes species in Panama. In light of the importance of these findings, further studies are needed focusing on the role of I. tapirus and I. cf. boliviensis as vectors, and the vertebrates acting as reservoirs.

A review of the genus Rickettsia in Central America.

In this paper, we present a historical review of rickettsiosis in Central America and also the most recent findings of Rickettsia in ectoparasites. All countries of Central America have records of rickettsiosis. Regarding the typhus group rickettsioses, there is clinical or serological evidence of Rickettsia prowazekii in Guatemala, Rickettsia typhi in Panama, Guatemala, and Costa Rica and unidentified species of the typhus group in El Salvador. Concerning spotted fever group rickettsiosis, there is serological evidence of infection by Rickettsia akari in Costa Rica and confirmed cases involving Rickettsia rickettsii in Panama and Costa Rica. There are also reports of spotted fever group rickettsiosis in acute patients from Guatemala, Honduras, and Nicaragua. Serological studies in Central America show reactivity of Rickettsia ambyommatis, Rickettsia bellii, Rickettsia felis, Rickettsia rhipicephali, and R. rickettsii in domestic and wild mammals. Eight species of Rickettsia have been detected in ectoparasites from Central America: R. africae (or very similar), R. amblyommatis, R. asembonensis, R. bellii, R. felis, R. parkeri, R. rhipicephali, and R. rickettsii, in addition to undescribed strains such as Atlantic Rainforest, Colombianensi, IbR/CRC, Barva, Aragaoi, and Candidatus "Rickettsia nicoyana;" the latter being the only one associated with Argasidae (Ornithodoros knoxjonesi). R. amblyommatis is the most common species in Central America, seeing as it has been reported in 10 species of ticks and one of fleas in five of the seven countries of the region. In this study, we demonstrate that the genus Rickettsia is widely distributed in Central America and that rickettsiosis could be an underestimated problem in the absence of greater diagnostic efforts in undetermined febrile cases.

Questing Amblyomma mixtum and Haemaphysalis juxtakochi (Acari: Ixodidae) Infected with Candidatus "Rickettsia amblyommii" from the Natural Environment in Panama Canal Basin, Panama.

This work emphasizes the detection of Candidatus "Rickettsia amblyommii" in questing Haemaphysalis juxtakochi and Amblyomma mixtum. From February 2009 to December 2012, questing ticks were collected from the vegetation and leaf-litter of four protected forests and two grassy areas around the Panama Canal basin. DNA was extracted from Amblyomma mixtum, Amblyomma naponense, Amblyomma oblongoguttatum, Amblyomma pecarium, Amblyomma tapirellum, Haemaphysalis juxtakochi, and unidentified immature Amblyomma. Specific primers of citrate synthase gene gltA were used to detect and identify the rickettsiae. Amplicons with the expected band size were purified and sequenced. DNA of C. "R. amblyommii" was found in A. mixtum, H. juxtakochi and Amblyomma immatures. To our knowledge, these finding represent the first report of C. "R. amblyommii" in free-living ticks in the wilderness of Central America.

Ticks (Ixodida) on humans from central Panama, Panama (2010-2011).

From January 2010 to December 2011, a total of 138 cases of ticks feeding on humans were reported from 11 locations in central Panama. Five of these locations were situated in forest environments, three in rural landscapes and three in urban areas. The ticks were submitted to the Gorgas Memorial Institute, where nine species were identified among 65 specimens: Amblyomma cajennense s.l., A. dissimile, A. naponense, A. oblongoguttatum, A. ovale, A. sabanerae, A. tapirellum, Haemaphysalis juxtakochi and Rhipicephalus sanguineus s.l. The remaining 73 specimens consisted of unidentified immature ticks, all belonging to the genus of Amblyomma. Rhipicephalus sanguineus s.l. was the species most frequently associated with humans, particularly in urban environments. In rural landscapes, tick bites were most often caused by A. cajennense s.l., whereas A. tapirellum was the species most often found parasitizing humans in forest environments. These data provide information on the tick species most commonly associated with humans in forested environments, rural areas and cities around the Panama Canal.

Rickettsial infection in domestic mammals and their ectoparasites in El Valle de Antón, Coclé, Panamá.

The present research evaluated the presence of Rickettsia spp. on ectoparasites of horses and dogs (using PCR techniques), and their sera (using immunofluorescence assay) in El Valle de Antón town in Panama. A total of 20 horses and 20 dogs were sampled, finding four species of ectoparasites on dogs (the ticks Rhipicephalus sanguineus, Amblyomma ovale, Amblyomma oblongoguttatum, and the flea Ctenocephalides felis), and two tick species on horses (Amblyomma cajennense and Dermacentor nitens). DNA of Rickettsia amblyommii was found in pools of A. cajennense, D. nitens, and R. sanguineus, while Rickettsia felis was detected in C. felis pools. Overall, 70% (14/20) and 65% (13/20) of the horses and dogs, respectively, were seroreactive (titer ≥ 64) to spotted fever group rickettsiae. Sera from six dogs and five horses reacted to R. amblyommii antigens with titers at least four-fold higher than those for the other antigens tested (Rickettsia bellii, Rickettsia parkeri, Rickettsia rhipicephali, R. felis, and R. rickettsii). These serological results, coupled with our molecular findings, suggest that these dogs and horses were infected by Rickettsia amblyommii. More studies need to be realized afford to identify the Rickettsia species responsible for other serological and molecular positive results, and their ecological importance.