Isolation and genetic characterization of a relapsing fever spirochete isolated from Ornithodoros puertoricensis collected in central Panama.

Tick-borne relapsing fever (TBRF) spirochetes are likely an overlooked cause of disease in Latin America. In Panama, the pathogens were first reported to cause human disease in the early 1900s. Recent collections of Ornithodoros puertoricensis from human dwellings in Panama prompted our interest to determine whether spirochetes still circulate in the country. Ornithodoros puertoricensis ticks were collected at field sites around the City of Panama. In the laboratory, the ticks were determined to be infected with TBRF spirochetes by transmission to mice, and we report the laboratory isolation and genetic characterization of a species of TBRF spirochete from Panama. Since this was the first isolation of a species of TBRF spirochete from Central America, we propose to designate the bacteria as Borrelia puertoricensis sp. nov. This is consistent with TBRF spirochete species nomenclature from North America that are designated after their tick vector. These findings warrant further investigations to assess the threat B. puertoricensis sp. nov. may impose on human health.

Free-living hard ticks (Ixodida: Ixodidae) from three different natural environments of Costa Rica.

This paper presents data on free-living ticks collected by flagging and using CO2 traps in three natural areas in Costa Rica: Carara National Park (CNP), Palo Verde National Park (PVNP), and a Private Forest Reserve in Sarapiquí (SPR). Data were analyzed calculating aspects of alpha diversity (species richness, entropy; dominance index, and evenness); and for beta diversity, compositional similarity between communities of ticks was also calculated. We collected 12,795 ticks belonging to 10 species: Amblyomma coelebs, Amblyomma dissimile, Amblyomma mixtum, Amblyomma naponense, Amblyomma cf. oblongoguttatum, Amblyomma cf. parvum, Amblyomma sabanerae, Amblyomma tapirellum, Haemaphysalis juxtakochi and Ixodes affinis. The number of species and individuals varied between sites: 5970 ticks were collected in CNP, 4443 in PVNP, and 2382 in SPR. Amblyomma cf. oblongoguttatum and A. cf. parvum were collected at all three sites, but A. mixtum was the most abundant species, even though it was not collected in SPR. Values of alpha diversity were calculated for CNP and SPR, while diversity in PVNP was the lowest of the three locations. Evenness was highest in SPR and lowest in CNP. The only community that presented dominance was PVNP. Beta diversity showed low similarity between the three locations with the lowest being CNP and SPR. For the three localities, estimates of the number of tick species based on presence/absence data was higher using flagging than CO2; and considering the stage of the ticks collected. More larvae were captured using CO2 traps than by flagging, while flagging was better for collecting adults. To our knowledge this is the first study in Costa Rica that compares these two sampling methods in three different environmental areas.

Diversity of Rickettsia in ticks collected from wild animals in Panama.

This paper presents new data about Rickettsia species detected in ticks collected from wild animals, using 16S rRNA, gltA and ompA. Rickettsia DNA was found in 66 of 101 ticks. Using EZ BioCloud libraries were produced reads that identified Rickettsia aeschlimannii, and Illumina BaseSpace produced reads of Rickettsia rickettsii group, Rickettsia bellii group, and unclassified Rickettsia. Using gltA and ompA gene-specific primers, R. aeschlimannii could not be confirmed, but detection of Rickettsia amblyommatis was achieved in Amblyomma auricularium, Amblyomma geayi, Amblyomma mixtum, and Amblyomma pacae; R. bellii from Amblyomma dissimile, "Candidatus Rickettsia colombianensi" from A. dissimile, Rickettsia spp. closely related to R. raoultii from A. geayi, Rickettsia tamurae from A. dissimile, and Rickettsia endosymbionts of Ixodes from Ixodes affinis. There were no databases available specifically for 16S rRNA of Neotropical Rickettsia, highlighting the need to use species primers over only 16S rRNA primers to achieve more accurate interpretations and identifications. These findings increase the number of Rickettsia species detected in Panama and highlight the need to establish isolates to further characterize the nature of Rickettsia in the area.

Morphological descriptions of the nymph and adults of Ornithodoros clarki, the larva and nymph of Ornithodoros rondoniensis, with notes on their phylogenetic relationships.

Based on tick specimens collected recently in Mexico, Nicaragua, Panama and Brazil, we provide morphological descriptions of the nymph and adults of Ornithodoros clarki Jones & Clifford, 1972 from the first three countries, and the larva and nymph of Ornithodoros rondoniensis (Labruna, Terassini, Camargo, Brandão, Ribeiro & Estrada-Peña, 2008) from Brazil. Also, an analysis of mitochondrial 16S rDNA sequences was performed to analyze the phylogenetic relationships of these tick species. Adults and nymphs of O. clarki and O. rondoniensis are unique among the Argasidae family by presenting exceptionally large spiracular plates with small goblets, and an integument with smooth polygonal mammillae. However, these two species are morphologically distinct based on specific patterns of coxal folds, idiosomal mammillae and pilosity, and female genital flap. In contrast, the larvae of O. clarki and O. rondoniensis are morphologically identical, except for a general larger size of the former species; this slight difference is corroborated by Principal Component Analysis (PCA) by using 40 morphometric variables. Phylogenetic analyses including 16S rDNA partial sequences of different Ornithodoros taxa from Central and South America indicate that O. rondoniensis from Brazil diverges from O. clarki from Mexico, Nicaragua and Panama. However, phylogenetic distance separating both alleged species is similar or slightly lower than the distances depicted for conspecific populations of a few other Ornithodoros species. Nonetheless, our primary criterion to maintain O. rondoniensis as a valid species is because its adult and nymphal stages do present distinct morphological traits that easily distinguish these postlarval stages from O. clarki.

Isolation of Rickettsia rickettsii in Rocky Mountain Spotted Fever Outbreak, Panama.

We report new cases of Rocky Mountain spotted fever in patients from Kinkantu, Ngäbe-Bugle indigenous comarca, Panama. We isolated Rickettsia rickettsii in cell culture after intraperitoneal inoculation of guinea pigs with tissues from a deceased patient. Our results indicate that Rocky Mountain spotted fever is emerging in this region.

Ornithodoros capensis sensu stricto (Ixodida: Argasidae) in Coiba National Park: first report for Panama, with notes on the O. capensis group in Panamanian shores and Costa Rica.

Ornithodoros capensis sensu lato (s.l.) is a morphologically similar group of soft ticks that parasitizes mostly seabirds in continental and offshore territories worldwide. Ornithodoros capensis sensu stricto (s.s.) has been previously recorded in many islands and coastal localities along the American continent; however, some records from Central America remain obscure. In this work we performed morphological and molecular analyses on soft ticks collected in Coiba National Park, an archipelago located in the Pacific Ocean off the coast of Panama, confirming the occurrence of O. capensis s.s. in this country for the first time. In addition, a morphological examination of museum specimens collected in Costa Rica, and a further locality in Panama, confirmed that O. capensis s.l. is established in the former country, and that its distribution along Panamanian shores is likely larger.

Hard tick (Acari: Ixodidae) survey of Oleoducto trail, Soberania National Park, Panama.

Hard tick diversity was determined along the Oleoducto trail (OT), Soberania National Park, from February 2013 to September 2014. Ticks were surveyed at four sites of 500 m2 each and with increasing forest cover gradient and decreasing disturbance. Tick collections were made by dragging and flagging vegetation, and traps and mist nets were used to capture mammals and birds. Animals confiscated from poachers were also examined. To obtain information about potential hosts along the trail, 20 camera traps were used. 1536 ticks were collected, representing 20 species; of these, 1089 were questing ticks (10 species) collected on flags. We examined 143 birds (30 species) and 59 mammals (10 species), of which 40 birds and 36 mammals had ticks. Site 1 presented the lowest number of species and also the lowest number of potential hosts. Artiodactyls were the most frequent mammals photographed in camera traps, and ticks that parasitize these animals were among the most abundant in sites 2-4. Of these, Haemaphysalis juxtakochi was the most abundant species. Differences among sites were consistent with the gradient of forest cover, disturbance along OT and distribution of potential hosts.

New records of Ornithodoros puertoricensis Fox 1947 (Ixodida: Argasidae) parasitizing humans in rural and urban dwellings, Panama.

The presence of ticks inside human constructions was evaluated in two localities from Colon province (Charco La Piedra and Espinar) and one from Panama province (Ancon, City of Panama). In two of houses, eight people from Charco La Piedra and one from Ancón reported "insect bites," which produced blisters for several weeks. The investigation resulted in the collection of argasid ticks, which were identified by morphology and sequencing the 16s ribosomal RNA gene, and later evaluated for the presence of relapsing fever Borrelia DNA. All ticks were identified as Ornithodoros puertoricensis. While spirochetal DNA was not detected by PCR in the ticks, our report highlights the potential for relapsing fever borreliosis in rural and urban localities in Panama.

First description of the male and redescription of the female of Ixodes tapirus Kohls, 1956 (Acari: Ixodidae), a parasite of tapirs (Perissodactyla: Tapiridae) from the mountains of Colombia, Costa Rica and Panama.

The male of Ixodes tapirus Kohls, 1956 (Acari: Ixodidae) is described for the first time and the female is redescribed in greater detail. Adults of I. tapirus are similar to those of Ixodes guatemalensis Kohls, 1956, Ixodes lasallei Méndez & Ortiz, 1958, Ixodes montoyanus Cooley, 1944 and Ixodes venezuelensis Kohls, 1953 but can be distinguished by their overall size, the amount of sclerotisation of the conscutum and accessory plates, the shape of the scutum, the number of punctations and their pattern on the conscutum and scutum, the depth of the punctations on the basis capituli dorsally, the shape and size of the porose areas and the size and shape of the auriculae. Adults of I. tapirus were collected from tapirs and vegetation in the mountains of Colombia, Panama and recorded from Costa Rica for the first time.