Abstract This report describes the finding of soft ticks (Ixodida, Argasidae) parasitizing two Andean flickers (Colaptes rupicola) from the Nuñoa district, Melgar province in Puno, Peru (14°31'11.77"S; 70°32'15.95''W; elevation 3967 m). A total of 29 larval ticks were collected directly from the birds and morphologically identified as Argas neghmei. Diagnosis was based on the larval morphology and confirmed by molecular analysis of the tick mitochondrial 16S rRNA gene. A sequence of 363 base pair of this gene showed to be 100% identical with sequences of A. neghmei from Argentina and Chile (GenBank: FJ853598 and DQ295781). This finding confirms the presence of A. neghmei in Peru and adds the Andean flicker as a new host for this species.

Resumen Este informe describe el hallazgo de garrapatas blandas (Ixodida, Argasidae) parasitando a dos carpinteros andinos (Colaptes rupicola) provenientes del distrito de Nuñoa, provincia de Melgar en Puno, Perú (14°31’11.77"S; 70°32’15.95"W; 3967 m de altitud). Un total de 29 larvas de garrapatas fueron colectadas directamente de las aves y posteriormente identificadas morfológicamente como Argas neghmei. El diagnóstico fue basado en la morfología de las larvas y confirmado mediante el análisis molecular del gen 16S ARNr mitocondrial de la garrapata. Un fragmento de 363 pares de bases de este gen mostró una identidad del 100% con secuencias registradas para A. neghmei de Argentina y Chile (GenBank: FJ853598 y DQ295781). Este hallazgo confirma la presencia de A. neghmei en Perú y agrega al carpintero andino como nuevo hospedero para esta garrapata.

Tick-borne rickettsioses are worldwide infectious diseases that are considered emerging and re-emerging. Until recently the only tick-borne rickettsiosis present in Latin America was Rickettsia rickettsii infection, but to date, with the incorporation of new tools as PCR and sequencing and the quick cellular close tube cultures (Shell-vial), new species has been involved as human pathogens. In these guidelines, we offer an update of the microbiological assays for diagnosing rickettsioses. Besides we have included a section in which the most important hard ticks involved in human rickettsioses in Latinoamerica are detailed.

Las rickettsiosis transmitidas por garrapatas son afecciones de distribución mundial, que por diferentes motivos se pueden considerar emergentes y reemergentes. Hasta hace escasos años la única rickettsiosis transmitida por garrapatas en Latinoamérica era la infección por Rickettsia rickettsii, pero en la actualidad y fundamentalmente, gracias a la incorporación de nuevas herramientas para el diagnóstico microbiológico como la reacción en cadena de la polimerasa y secuenciación o el cultivo celular rápido en tubo cerrado, se han descrito e involucrado otras especies de Rickettsia en la producción de patología humana. En estas guías se detallan y describen las diferentes técnicas utilizadas para el diagnóstico microbiológico de las rickettsiosis. Además, se incluye una sección en la que se detallan las especies más importantes de garrapatas duras relacionadas con las rickettsiosis en Latinoamérica, con claves para su clasificación taxonómica.

Rangelia vitalii é um protozoário que causa doença em cães, sendo a anemia o achado laboratorial mais frequente. No entanto, existem poucos estudos sobre as alterações bioquímicas em cães infectados com o protozoário. Assim, este estudo tem como objetivo investigar as alterações bioquímicas de cães experimentalmente infectados com R. vitalii na fase aguda da infecção. Para o estudo, foram utilizados 12 cães fêmeas (com idade entre 6 a 12 meses e peso entre 4 a 7 kg), divididos em dois grupos. O grupo A (n = 5) foi composto de animais saudáveis e o grupo B (n = 7) de animais infectados. Amostras de sangue foram coletadas nos dias zero, dez, vinte e trinta PI, utilizando tubos sem anticoagulante para obtenção de soro e análise dos parâmetros bioquímicos. Foi observado um aumento na alanino aminotransferase (ALT) no dia 20 PI (P < 0,05) e aumento na creatinoquinase (CK) e aspartato aminotransferase (AST) em todo o período experimental (P < 0,05). Não foram observadas alterações séricas na gama-glutamiltransferase, uréia e creatinina. Portanto, é possível concluir que a infecção experimental por R. vitalii causa alterações no perfil bioquímico, com aumento na ALT, CK e AST.

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.

Introducción. Los capibaras o chigüiros (Hydrochoerus hydrochaeris) son huéspedes amplificadores de Rickettsia sp. Usualmente se encuentran parasitados por la garrapata Amblyomma cajennense, principal vector de rickettsiosis en Suramérica. Los capibaras pueden ser usados como potenciales centinelas de la circulación de rickettsias. Objetivo. Detectar anticuerpos contra Rickettsia sp. del grupo de las fiebres manchadas en capibaras de una zona rural del municipio de Montería, departamento de Córdoba. Material y métodos. Se analizaron 36 sueros de capibaras de una zona rural de Montería (vereda San Jerónimo) en Córdoba. Para la detección de anticuerpos IgG se practicó inmunofluorescencia indirecta, que utilizó antígenos de la cepa Taiaçu de Rickettsia rickettsii de Brasil. Los sueros de los capibaras fueron diluidos 1:64. Se capturaron las garrapatas que se encontraban parasitando los capibaras y se clasificaron hasta su especie. Resultados. La seroprevalencia contra Rickettsia sp. del grupo de la fiebres manchadas encontrada fue de 22 % (8 capibaras); se encontraron cuatro sueros con título de 1:64, tres sueros con título 1:128 y un suero presentó titulación de 1:512. Todas las garrapatas (n=933) fueron identificadas taxonómicamente como A. cajennense. Conclusión. En Colombia existen zonas endémicas de rickettsiosis y la aparición de brotes anuales lo confirma (Necoclí, 2006; Los Córdobas, 2007, y Altos de Mulatos, 2008). El presente estudio reporta por primera vez la presencia de infección natural por rickettsia del grupo de las fiebres manchadas en capibaras de Colombia. Los hallazgos sugieren que los capibaras pueden ser usados como potenciales centinelas de la circulación de rickettsias y marcadores de las áreas de riesgo para la transmisión de rickettsiosis.

Introduction. Capybaras (Hydrochoerus hydrochaeris) are considered amplifying hosts of Rickettsia sp. These rodents are usually parasitized by the tick vector, Amblyomma cajennense, the main vector of rickettsioses in humans and animals in South America. Capybaras can be used as sentinels in detection of circulation of rickettsiae. Objective. Antibodies to rickettsiae of spotted fever group were detected in capybaras in a rural area of Cordoba Province, northern Colombia. Materials and methods. Sera were analyzed from 36 capybaras in a rural area of Monteria (village of San Jeronimo) in Córdoba. For the detection of IgG antibodies, indirect immunofluorescence was performed. The antigens were derived from R. rickettsia strain Taiaçu isolated in Brazil. Capybara sera were diluted 1:64 for IFA analysis. Ticks were collected from each capybara (also known as chigüiro) and identified to species. Results. The seroprevalence of spotted fever group Rickettsia was 22% (8 capybaras). Four sera had a titer of 1:64, 3 had a titer of 1:128 and one serum had a titer of 1:512. All ticks removed from the capybaras (n=933) were taxonomically identified as Amblyomma cajennense. Conclusion. Colombia has areas endemic for rickettsioses, as indicated by confirmed annual outbreaks. The current study reports the first evidence of natural rickettsial infection of the spotted fever group in capybaras from Colombia. The findings suggest that capybaras can be used as sentinels for the circulation of rickettsiae and can identify endemic areas for the transmission of rickettsial diseases.

Reportes del genero Rickettsia y sus asociadas infecciones fueron compilados en una revisión crítica retrospectiva de la literatura científica de los países de Latinoamérica, el Caribe, Portugal y España. Se consideraron todos los reportes para huéspedes humanos y/o animales y también para huéspedes invertebrados los cuales fueron considerados como vectores asociados con Rickettsia. En algunos casos, cuando no existió detección directa a un determinado grupo de Rickettsias o especies no disponible en un país, se tuvo en cuenta la detección indirecta por serología. Un total de 13 especies de Rickettsia han sido reportadas en Latinoamérica y el Caribe. Las especies más encontradas en los países fueron: Rickettsia felis (9 países), R. prowazekii (7 países), R. typhi (6 países), R. rickettsii (6 países), R. amblyommii (5 países) y R. parkeri (4 países). Los datos de las islas del Caribe (antillas menores o Indias occidentales), fueron agrupados en una sola área geográfica como un solo país. Ambas R. bellii, R. akari y Candidatus 'R. andeane' fueron reportadas en solo 2 países, mientras que R. massiliae, R. rhipicephali, R.monteiroi, y R. africae fueron informadas en un solo país. En este caso R. africae fue reportada en 9 islas de las Antillas menores. Para El Salvador, Honduras y Nicaragua, hasta ahora no se han reportado especies de Rickettsia, pero si evidencia serológica de infección humana y/o animal. Sin reportes de infección por Rickettsia permanecen: Belice, Venezuela, Guayana, Surinam y Paraguay. Además, a excepción de algunas islas del Caribe, muchas de ellas permanecen sin reportes. Un total de 12 especies de Rickettsia han sido documentadas en España y Portugal: R. conorii, R. helvetica, R. monacensis, R. felis, R. slovaca, R. raoultii, R. sibirica, R. aeschlimannii, R. rioja, R. massiliae, R. typhi y R. prowazekii. Entre estas, solamente R. prowazekii, R. typhi, R. felis y R. massiliae han sido documentados en Latinoamérica, España y Portugal. Los datos de este estudio permiten entender mejor la epidemiología de las Rickettsias en Latinoamérica, Caribe, España y Portugal, y la distribución de los vectores.

Data on genus and infectious by Rickettsia were retrospectively compiled from the critical review literature regarding all countries in Latin America, Caribbean islands, Portugal and Spain. We considered all Rickettsia records reported for human and/or animal hosts, and/or invertebrate hosts considered being the vector. In a few cases, when no direct detection of a given Rickettsia group or species was available for a given country, the serologic method was considered. A total of 13 Rickettsia species have been recorded in Latin America and the Caribbean. The species with the largest number of country confirmed records were Rickettsia felis (9 countries), R. prowazekii (7 countries), R. typhi (6 countries), R. rickettsii (6 countries), R. amblyommii (5 countries), and R. parkeri (4 countries). The Rickettsial records for the Caribbean islands (West Indies) were grouped in only one geographical area. Both R. bellii, R. akari, and Candidatus 'R. andeane' have been recorded in only 2 countries each, whereas R. massiliae, R. rhipicephali, R.monteiroi, and R. africae have each been recorded in a single country (in this case, R. africae has been recorded in nine Caribbean Islands). For El Salvador, Honduras, and Nicaragua, no specific Rickettsia has been reported so far, but there have been serological evidence of human or/and animal infection. The following countries remain without any Rickettsial records: Belize, Venezuela, Guyana, Surinam, and Paraguay. In addition, except for a few islands, many Caribbean islands remain without records. A total of 12 Rickettsia species have been reported in Spain and Portugal: R. conorii, R. helvetica, R. monacensis, R. felis, R. slovaca, R. raoultii, R. sibirica, R. aeschlimannii, R. rioja, R. massiliae, R. typhi, and R. prowazekii. Amongst these Rickettsia species reported in Spain and Portugal, only R. prowazekii, R. typhi, R. felis, and R. massiliae have also been reported in Latin America. This study summarizes the current state of art on the Rickettsial distribution in Latin America, Caribbean, Spain and Portugal. The data obtained allow a better understanding on Rickettsial epidemiology and distribution of vector ecology.

Amblyomma scalpturatum Neumann has been reported in Brazil in the northern (States of Amazonas, Pará and Rondônia) and mid-western regions (States of Mato Grosso and Mato Grosso do Sul). It has been found in association with large and medium sized mammals, as Artiodactyla, Perissodactyla, Xenarthra and Carnivora. In this paper, this tick species is reported for the first time in the Brazilian states of Paraná and Roraima, southern and northern regions, respectively.

The bacterium Rickettsia rickettsii is the etiological agent of an acute, severe disease called Rocky Mountain spotted fever in the United States or Brazilian spotted fever (BSF) in Brazil. In addition to these two countries, the disease has also been reported to affect humans in Mexico, Costa Rica, Panama, Colombia and Argentina. Like humans, dogs are also susceptible to R. rickettsii infection. However, despite the wide distribution of R. rickettsii in the Western Hemisphere, reports of R. rickettsii-induced illness in dogs has been restricted to the United States. The present study evaluated the pathogenicity for dogs of a South American strain of R. rickettsii. Three groups of dogs were evaluated: group 1 (G1) was inoculated ip with R. rickettsii; group 2 (G2) was infested by R. rickettsii-infected ticks; and the control group (G3) was infested by uninfected ticks. During the study, no clinical abnormalities, Rickettsia DNA or R. rickettsii-reactive antibodies were detected in G3. In contrast, all G1 and G2 dogs developed signs of rickettsial infection, i.e., fever, lethargy, anorexia, ocular lesions, thrombocytopenia, anemia and detectable levels of Rickettsia DNA and R. rickettsii-reactive antibodies in their blood. Rickettsemia started 3-8 days after inoculation or tick infestation and lasted for 3-13 days. Our results indicate that a Brazilian strain of R. rickettsii is pathogenic for dogs, suggesting that canine clinical illness due to R. rickettsii has been unreported in Brazil and possibly in the other South American countries where BSF has been reported among humans.

Objetivo. Aislar Rickettsias mediante cultivo celular a partir de muestras de garrapatas Amblyomma ovale y Amblyomma incisum del estado de São Paulo Materiales y métodos. A. ovale y A. incisum adultas de vida libre fueron colectadas en una área de selva tropical Atlántica en el estado de São Paulo, Brazil. Cada garrapata fue sometida a la prueba de hemolinfa, las garrapatas positivas en esta prueba fueron evaluadas con la técnica de shell vial con el propósito de aislar rickettsias en cultivo de células Vero. Pasajes celulares de los aislados fueron identificados genotípicamente por la reacción en cadena por la polimerasa (PCR) dirigidos a fragmentos de tres genes de rickettsias (gltA, htrA y ompA), seguido por secuenciación de ADN. Resultados. Un total de 388 A. incisum y 50 A. ovale fueron colectadas. Por la prueba de hemolinfa, únicamente una A. incisum y una A. ovale fueron positivas. Las Rickettsias fueron exitosamente aisladas de estas garrapatas. Sin embargo, el cultivo continuo en células Vero fue posible sólo para la garrapata A. ovale, debido a contaminación bacteriana en el primer pasaje celular de la muestra de A. incisum. Los productos de PCR fueron obtenidos con los primers gltA y htrA para los dos aislados, no obstante, ningún producto fue obtenido con los primers ompA. Por análisis BLAST, secuencias parciales de gltA y htrA procedentes de los aislados de A. ovale y A. incisum fueron similares a las secuencias correspondientes a R. bellii. Conclusiones. Este es el primer reporte de R. bellii infectando A. incisum y el primer establecimiento exitoso de un aislado de A. ovale.

Objective. To isolate and characterize rickettsiae from the ticks Amblyomma ovale and Amblyomma incisum collected in the state of São Paulo. Materials and methods. Adult, free-living A. ovale and A. incisum were collected in an Atlantic rainforest area in the state of São Paulo, Brazil. Each tick was tested using the hemolymph assay; samples from positive ticks were placed in shell vials in order to isolate rickettsiae and subsequently grown in Vero cells. Amplification of three rickettsial genes (gltA, htrA and ompA) was attempted using polymerase chain reaction (PCR) for each isolate obtained. Amplicons were subsequently sequenced. Results. A total of 388 A. incisum and 50 A. ovale were collected. Only one A. incisum and one A. ovale were hemolymph-test positive. Rickettsiae were successfully isolated from these ticks; however establishment in Vero cell culture was successful only for the isolate from A. ovale. Bacterial contamination in the first cell passage of the A. incisum isolate precluded successful isolation of the organism. PCR products were obtained with the gltA and htrA primers for the two isolates, but no product was obtained with the ompA primers. By BLAST analysis, partial gltA and htrA sequences of isolates from A. ovale and A. incisum were similar to the corresponding sequences of R. bellii. Conclusions. This is the first report of R. bellii infecting A. incisum and the first successful isolation from A. ovale.

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.

Introducción. En Brasil, los capibaras (Hydrochoerus hydrochaeris) son importantes huéspedes para garrapatas del género Amblyomma, las cuales transmiten rickettsiosis a humanos y animales. Por lo tanto, estos roedores pueden ser potenciales centinelas para detectar infección por rickettsia. Objetivos. Este trabajo evaluó la infección por rickettsia en capibaras de diferentes regiones del estado de São Paulo, donde las rickettsiosis nunca han sido reportadas. Materiales y métodos. Se examinarion los sueros de 73 capibaras de seis localidades en São Paulo con la prueba de immunofluorescencia indirecta con antígenos de Rickettsia rickettsii, Rickettsia parkeri y Rickettsia bellii. Los bazos de los capibaras se extrajeron y se analizaron por reacción en cadena de la polimerasa para un fragmento del gene gltA de rickettsia. Las garrapatas se recolectaron de los capibaras y se identificaron hasta especie. Resultados. Diecinueve (26,0%), 25 (34,2%) y 50 (68,5%) sueros de los capibaras reaccionaron con R. rickettsii, R. parkeri y R. bellii, respectivamente. De los 50 sueros que reaccionaron con antígenos de R. bellii, 25 presentaron títulos, por lo menos, cuatro veces mayores que los otros dos antígenos. Estos sueros fueron considerados homólogos de R. bellii. Usando el mismo criterio, tres sueros de los capibaras se consideraron homólogos de R. parkeri. Ningún suero se consideró homólogo de R. rickettsii. No se detectó ADN de rickettsia en bazo. Las garrapatas recolectadas de los capibaras fueron identificadas como Amblyomma dubitatum y Amblyomma cajennense. Conclusiones. Este trabajo reporta la primera evidencia de infección natural por R. bellii en vertebrados y, también, la primera evidencia de infección por R. parkeri en capibaras. Se sabe que R. parkeri infecta y produce enfermedad en humanos; sin embargo, no hay evidencia de infección humana por R. bellii.

Introduction. In Brazil, capybaras (Hydrochoerus hydrochaeris) are important hosts for Amblyomma ticks, which in turn can transmit rickettsiae to humans and animals. Therefore, capybaras are potential sentinels for rickettsial infection. Objective. The present study evaluated rickettsial infection in capybaras in different areas of the state of São Paulo, where rickettsiosis has never been reported. Materials and methods. Blood sera from 73 capybaras from six localities in São Paulo were tested by indirect immunofluorescence assay using Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia bellii antigens. Capybara spleens were tested by PCR, targeting a fragment of the rickettsial gltA gene. Ticks were collected from each capybara sample and taxonomically identified to species. Results. A total of 94 positively reacting capybara samples, 19 (26.0%), 25 (34.2%), and 50 (68.5%) capybara sera reacted to R. rickettsii, R. parkeri, and R. bellii, respectively. Twenty-five capybara sera showed titers to R. bellii at least four-fold higher than to any of the other two antigens. These sera were considered homologous to R. bellii. Using the same criteria, 3 capybara sera were considered homologous to R. parkeri. No sera were be considered homologous to R. rickettsii. No rickettsial DNA was detected in capybara spleen samples. Ticks collected on capybaras were Amblyomma dubitatum and Amblyomma cajennense. Conclusions. The first evidence is reported of R. bellii natural infection in vertebrate hosts, and the first evidence of R. parkeri infection in capybaras. While R. parkeri is known to infect and cause disease in humans, no similar evidence for human infection has been indicated by R. bellii.

The present study evaluated rickettsial infection in Amblyomma spp. ticks collected in a farm in Coronel Pacheco, a Brazilian spotted fever (BSF) endemic area. A total of 78 A. cajennense and 78 A. dubitatum free-living adult ticks were collected and tested by polymerase chain reaction (PCR) targeting a fragment of the rickettsial gene gltA. Only one pool of three A. cajennense ticks showed the expected product by PCR. This pool was further tested by PCR using sets of primers targeting the rickettsial genes gltA, ompA, and ompB. All reactions yielded the expected bands that by sequencing, showed 100% identity to the corresponding sequences of the Rickettsia rickettsii gene fragments gltA (1063-bp), ompA (457-bp), and ompB (720-bp). The minimal infection rate of R. rickettii in the A. cajennense population was 1.28% (at least one infected tick within 78 ticks).The present study showed molecular evidence for the presence of R. rickettsii in A. cajennense from a BSF-endemic area in Coronel Pacheco, state of Minas Gerais. Although R. rickettsii has been previously reported infecting A. cajennense ticks in Brazil and other Latin American countries, the present study performed the first molecular characterization of R. rickettsii from the tick A. cajennense.

From June 2000 to June 2001, a total of 741 ticks were collected from 51 free-living wild animals captured at the Porto-Primavera Hydroelectric power station area, located alongside an approximately 180 km course of the Paraná river, between the states of São Paulo and Mato Grosso do Sul, comprising 9 species of 3 genera: Ambly-omma (7 species), Boophilus (1) and Anocentor (1). A total of 421 immature Amblyomma ticks were reared in laboratory until the adult stage, allowing identification of the species. A. cajennense was the most frequent tick species (mostly immature stages) collected on 9 host species: Myrmecophaga tridactyla, Tamandua tetradactyla,Cerdocyon thous, Puma concolor,Tayassu tajacu, Mazama gouazoubira,Hydrochaeris hydrochaeris,Alouatta caraya, Cebus apella. Other tick species were less common, generally restricted to certain host taxa.