Candidatus Rickettsia andeanae, a spotted fever group agent infecting Amblyomma parvum ticks in two Brazilian biomes

Adult ticks of the species Amblyomma parvum were collected from the vegetation in the Pantanal biome (state of Mato Grosso do Sul) and from horses in the Cerrado biome (state of Piauí) in Brazil. The ticks were individually tested for rickettsial infection via polymerase chain reaction (PCR) targeting three rickettsial genes, gltA, ompA and ompB. Overall, 63.5% (40/63) and 66.7% (2/3) of A. parvum ticks from Pantanal and Cerrado, respectively, contained rickettsial DNA, which were all confirmed by DNA sequencing to be 100% identical to the corresponding fragments of the gltA, ompA and ompB genes of Candidatus Rickettsia andeanae. This report is the first to describe Ca. R. andeanae in Brazil.

Rickettsia amblyommii infecting Amblyomma sculptum in endemic spotted fever area from southeastern Brazil

The Rickettsia bacteria include the aetiological agents for the human spotted fever (SF) disease. In the present study, a SF groupRickettsia amblyommii related bacterium was detected in a field collected Amblyomma sculptum (Amblyomma cajennense species complex) tick from a Brazilian SF endemic site in southeastern Brazil, in the municipality of Juiz de Fora, state of Minas Gerais. Genetic analysis based on genes ompA,ompB and htrA showed that the detected strain, named R. amblyommii str. JF, is related to the speciesR. amblyommii.

Análise do número de cerdas internas do órgão de Haller de larvas de Amblyomma nodosum Neumann (Acari, Ixodidae)

A partir de teleógina de Amblyomma nodosum Neumann, 1899, removida de Tamandua tetradactyla (Linnaeus, 1758) foram obtidas posturas que originaram larvas, em condições de laboratório. Um total de 219 larvas foi examinado em microscopia de luz, com objetiva de imersão e microscopia de confocal com varredura a laser. Buscou-se analisar a variação do número das cerdas internas dos lados direito e esquerdo, no fundo da cápsula do Órgão de Haller, com intuito de utilizá-las como um parâmetro quetotáxico na diagnose específica. A média do número de cerdas foi 5,06±0, 71, com valores limites da amplitude de 4-11.

Patterns of parasitism by Amblyomma rotundatum (Ixodidae) on Rhinella schneideri (Bufonidae) in islands of São Francisco River, Minas Gerais, Brazil

Abstract: We sampled ticks from specimens of the rococo toad Rhinella schneideriby flannel dragging on two Islands located in the São Francisco River near the Três Marias hydroelectric dam, southeastern Brazil. A total of 120 toads was examined, of which 63 (52.5%) were parasitized only by Amblyomma rotundatumtotaling 96 larvae, 163 nymphs and 134 females. The burden of parasitism ranged from one to 43 ticks, with a mean intensity of infestation of 6.2±5.5 ticks per host. The tick A. rotundatumexhibited highly aggregated distribution. Peak abundance of larvae and nymphs occurred in the dry season (May to September), whereas peak abundance of females occurred in the wet season (October to April). We collected most ticks near the head and hind limbs of R. schneideri. The finding of two engorged A. rotundatumnymphs in the same resting places of two toads and the absence of this species in the dragged areas suggest a nidicolous behavior at the studied site.

Parasitos de aves e mamíferos silvestres em cativeiro no estado de Pernambuco

Resumo: Os animais silvestres são hospedeiros de uma grande variedade de parasitos que podem interferir em sua conservação ex situ. O objetivo deste estudo foi identificar os parasitos gastrointestinais (PGI) e ectoparasitos dos animais do Centro de Triagem de Animais Silvestres (CETAS) do Instituto Brasileiro de Meio Ambiente e Recursos Naturais Renováveis (IBAMA) de Recife, Pernambuco, além de determinar os aspectos do manejo em cativeiro que possam estar relacionados com os parasitos identificados. Foram coletados ectoparasitos e amostras fecais de 223 aves e mamíferos, as quais foram processadas pelos métodos: microscopia direta, flutuação e sedimentação. Helmintos e/ou protozoários foram detectados em 91 (40,8%) amostras fecais, sendo 64 (70,3%) de aves e 27 (29,7%) de mamíferos. Ovos de Capillaria sp., Ascaridida, Spirurida e oocistos de Eimeria sp. foram detectados nas amostras fecais das aves, enquanto ovos de Trichuris trichiura, Strongyloides sp., Toxocara canis, Ancylostoma sp., Strongylida e oocistos de Coccídios foram encontrados nas amostras fecais de mamíferos. Os ectoparasitos identificados em aves foram Colpocephalum turbinatum, Kurodaia (Kurodaia) fulvofasciata, Halipeurus sp., Naubates sp., Saemundssonia sp., Austromenopon sp., Paragoniocotes sp., Brueelia sp., Myrsidea sp. and Pseudolynchia sp., enquanto em mamíferos os ectoparasitos identificados foram Rhipicephalus sanguineus, Amblyomma varium, A. calcaratum, A. nodosum, Ornithodoros talaje e Ctenocephalides felis felis. A. calcaratum e O. talaje são registrados pela primeira vez em Pernambuco e T. tetradactyla é apresentado como novo hospedeiro de O. talaje. Nenhum dos animais estudados apresentou sinais clínicos em decorrência da infecção/infestação parasitária. Parasitos com potencial zoonótico como T. trichiura, Strongyloides sp., T. canis e Ancylostoma sp. foram identificados em primatas não humanos e carnívoros. Precárias condições estruturais e sanitárias do CETAS-PE estão relacionadas com os parasitos identificados neste estudo e devem ser levadas em consideração para a adoção de medidas adequadas de controle. Os resultados deste estudo contribuirão de maneira significativa para a conservação de animais selvagens no CETAS-PE e para a saúde dos profissionais responsáveis pela manutenção destes animais.

Rickettsiosis cutáneo ganglionar por Rickettsia conorii en el Uruguay

Se refieren 3 casos autóctonos de rickettsiosis cutáneo ganglionar trasmitidos por garrapatas de perros (Amblyomma maculatum, en uno de ellos) en el Uruguay. Dos de los 3 casos fueron seguramente provocados por Rickettsia conorii de acuerdo a los resultados de la reacción específica de inmunofluorescencia indirecta - IgM, anti R. conorii. Se incluye un tercer paciente no estudiado con tal técnica, por la similitud clínico-epidemiológica, la reactividad del suero frente al Proteus OX 19 y la rápida respuesta a la tetraciclina. La no descripción previa de la rickettsiosis por R. conorii en forma autóctona en el área de las Américas confiere especial interés a la comunicación, recomendándose la búsqueda de la afección en otros países de la región.

Evidence of spotted fever group rickettsiae in state of Rio de Janeiro, Brazil

Ticks were obtained from dogs from February to September of 1999 at weekly intervals, in the County of Piraí, State of Rio de Janeiro. Four hundred seventy four ixodids were taxonomically identified, 103 Amblyomma cajennense, seven Amblyomma ovale, 209 Rhipicephalus sanguineus, and 155 Amblyomma sp. An hemolymph test associated with Giemsa's stain revealed two specimens in 163 ticks tested (R. sanguineus and Amblyomma sp), containing rickettsia-like organisms. Direct immunofluorescence verified the presence of spotted fever group rickettsia in one specimen of R. sanguineus. Considering the limited information on rickettsiosis in Brazil, principally in relation to the vectors involved in perpetuating it in foci, these preliminary results give us an idea on the importance of infection in ticks, allowing to expand our knowledge on this zoonosis.

Rickettsial spotted fever in capoeirão Village, Itabira, Minas Gerais, Brazil

The present study investigated the infection by spotted fever rickettsia in an endemic area for Brazilian spotted fever (BSF; caused by Rickettsia rickettsii) in Minas Gerais State, Brazil. Human, canine and equine sera samples, and Amblyomma cajennense adult ticks collected in a rural area of Itabira City, Minas Gerais State were tested for rickettsial infection. Through Immunofluorescence Assay (IFA) we demonstrated the presence of antibodies anti-R. rickettsii in 8.2%, 81.3% and 100% of the human, canine and equine sera, respectively. None of the 356 tick specimens analyzed were positive for Rickettsia by the hemolymph test or Polymerase Chain Reaction technique (PCR) for the htrA and the gltA genes. Our serological results on horses and dogs (sentinels for BSF) appoint for the circulation of a SFG Rickettsia in the study area, however in a very low infection rate among the A. cajennense tick population.

Rickettsia parkeri: a Rickettsial pathogen transmitted by ticks in endemic areas for spotted fever rickettsiosis in southern Uruguay

At first Rickettsia conorii was implicated as the causative agent of spotted fever in Uruguay diagnosed by serological assays. Later Rickettsia parkeri was detected in human-biting Amblyomma triste ticks using molecular tests. The natural vector of R. conorii, Rhipicephalus sanguineus, has not been studied for the presence of rickettsial organisms in Uruguay. To address this question, 180 R. sanguineus from dogs and 245 A. triste from vegetation (flagging) collected in three endemic localities were screened for spotted fever group (SFG) rickettsiosis in southern Uruguay. Tick extracted DNA pools were subjected to PCR using primers which amplify a fragment of the rickettsial gltA gene. Positive tick DNA pools with these primers were subjected to a second PCR round with primers targeting a fragment of the ompA gene, which is only present in SFG rickettsiae. No rickettsial DNA was detected in R. sanguineus. However, DNA pools of A. triste were found to be positive for a rickettsial organism in two of the three localities, with prevalences of 11.8% to 37.5% positive pools. DNA sequences generated from these PCR-positive ticks corresponded to R. parkeri. These findings, joint with the aggressiveness shown by A. triste towards humans, support previous data on the involvement of A. triste as vector of human infections caused by R. parkeri in Uruguay.

SEROLOGICAL SURVEY OF Ehrlichia SPECIES IN DOGS, HORSES AND HUMANS: ZOONOTIC SCENERY IN A RURAL SETTLEMENT FROM SOUTHERN BRAZIL

SUMMARY The aims of this study were to determine the seroprevalence of Ehrlichia spp. and risk factors for exposure in a restricted population of dogs, horses, and humans highly exposed to tick bites in a Brazilian rural settlement using a commercial ELISA rapid test and two indirect immunofluorescent assays (IFA) with E. canis and E. chaffeensis crude antigens. Serum samples from 132 dogs, 16 horses and 100 humans were used. Fifty-six out of 132 (42.4%) dogs were seropositive for E. canis. Dogs > one year were more likely to be seropositive for E. canis than dogs ≤ one year (p = 0.0051). Ten/16 (62.5%) and 8/16 (50%) horses were seropositive by the commercial ELISA and IFA, respectively. Five out of 100 (5%) humans were seropositive for E. canis and E. chaffeensis. Rhipicephalus sanguineus (n = 291, 97.98%) on dogs and Amblyomma cajennense (n = 25, 96.15%) on horses were the most common ticks found. In conclusion, anti-Ehrlichia spp. antibodies were found in horses; however, the lack of a molecular characterization precludes any conclusion regarding the agent involved. Additionally, the higher seroprevalence of E. canis in dogs and the evidence of anti-Ehrlichia spp. antibodies in humans suggest that human cases of ehrlichiosis in Brazil might be caused by E. canis, or other closely related species.

Epidemiological aspects of the Brazilian spotted fever: seasonal activity of ticks collected in an endemic area in São Paulo, Brazil

Ticks were collected from vegetation and animals at monthly intervals during one year (1993-1994) in an endemic area of Brazilian spotted fever in the County of Pedreira, State of São Paulo. Six species of ticks were identified Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus and Boophilus microplus. Only the first species was sufficiently numerous to permit a quantitative study with seasonal activity, although the distribution and source of capture of other species were observed and are reported. This information is correlated with the epidemiology of tick-borne rickettsiosis.

Isolamento de Rickettsia em cultura de células vero

Embora o diagnóstico da febre maculosa baseie-se em sinais e sintomas característicos, o mesmo requer confirmação laboratorial, pois existem alguns diagnósticos diferenciais possíveis como meningococcemia, leptospirose, infecção por enterovírus e febre tifóide. A confirmação laboratorial pode ser feita através da pesquisa de anticorpos específicos, possível somente alguns dias após o aparecimento da doença, através do isolamento do agente em amostras de sangue e/ou biópsia de pele, e ainda, de amostras de carrapatos coletados do paciente ou de animais reservatório. O isolamento a partir de sangue ou biópsia de pele resulta em diagnóstico precoce da doença, pois na fase de rickettsemia ainda não há anticorpos detectáveis no sangue. Assim, com o objetivo de facilitar o diagnóstico precoce da febre maculosa, estabelecemos um método de isolamento de rickettsia em cultura de células vero. Para a padronização foi inoculada amostra padrão de Rickettsia rickettsii, cepa Sheyla Smith, cedida pelo CDC. A identificação foi feita através da reação de imunofluorescência indireta. A presença de microrganismos verdes fluorescentes visualizados no interior do citoplasma das células caracterizou o crescimento do agente. Posteriormente, a metodologia foi confirmada pelo isolamento do agente da febre maculosa em amostras de biópsia de pele de paciente proveniente de área endêmica no Estado de São Paulo, bem como, de amostras de carrapato do gênero Amblyomma, considerado o reservatório e transmissor da doença no Brasil.

Report on ticks collected in the Southeast and Mid-West regions of Brazil: analyzing the potential transmission of tick-borne pathogens to man

Specimens of ticks were collected in 1993, 1996, 1997, and 1998, mostly from wild and domestic animals in the Southeast and Mid-West regions of Brazil. Nine species of Amblyommidae were identified: Anocentor nitens, Amblyomma cajennense, Amblyomma ovale, Amblyomma fulvum, Amblyomma striatum, Amblyomma rotundatum, Boophilus microplus, Boophilus annulatus, and Rhipicephalus sanguineus. The potential of these tick species as transmitters of pathogens to man was analyzed. A Flaviviridade Flavivirus was isolated from Amblyomma cajennense specimens collected from a sick capybara (Hydrochaeris hydrochaeris). Amblyomma cajennense is the main transmitter of Rickettsia rickettsii (=R. rickettsi), the causative agent of spotted fever in Brazil. Wild mammals, mainly capybaras and deer, infested by ticks and living in close contact with cattle, horses and dogs, offer the risk of transmission of wild zoonosis to these domestic animals and to man.

Study of infection by Rickettsiae of the spotted fever group in humans and ticks in an urban park located in the City of Londrina, State of Paraná, Brazil

INTRODUCTION: Spotted fevers are emerging zoonoses caused by Rickettsia species in the spotted fever group (SFG). Rickettsia rickettsii is the main etiologic agent of Brazilian spotted fever (BSF) and it is transmitted by Amblyomma spp. ticks. METHODS: The study aimed to investigate SFG rickettsiae in the Arthur Thomas Municipal Park in Londrina, PR, by collecting free-living ticks and ticks from capybaras and blood samples from personnel working in these areas. Samples from A. dubitatum and A. cajennense were submitted for PCR in pools to analyze the Rickettsia spp. gltA (citrate synthase gene). RESULTS: All the pools analyzed were negative. Human sera were tested by indirect immunofluorescence assay with R. rickettsii and R. parkeri as antigens. Among the 34 sera analyzed, seven (20.6%) were reactive for R. rickettsii: four of these had endpoint titers equal to 64, 2 titers were 128 and 1 titer was 256. None of the samples were reactive for R. parkeri. An epidemiological questionnaire was applied to the park staff, but no statistically significant associations were identified. CONCLUSIONS: The serological studies suggest the presence of Rickettsiae related to SFG that could be infecting the human population studied; however, analysis of the ticks collected was unable to determine which species may be involved in transmission to humans.

Risk factors associated with the transmissionof Brazilian spotted fever in the Piracicaba river basin, State of São Paulo, Brazil

INTRODUCTION : Brazilian spotted fever (BSF) is a disease transmitted by ticks for which the etiological agent is Rickettsia rickettsii. The present essay evaluates the risk factors associated with the transmission of cases of BSF in the time period between 2003 and 2013 in the Piracicaba river basin, state of São Paulo. METHODS : This essay presents a retrospective study to identify the factors associated with the transmission of cases of BSF among all suspected cases identified by the System for Epidemiological Surveillance of São Paulo (CVE). After the description of temporal distribution (onset of symptoms) and the environmental and demographic variations of the confirmed and discarded cases, a multiple logistic regression model was applied. RESULTS : We searched 569 probable locations of infection (PLI) with 210 (37%) confirmed cases of BSF and 359 (63%) discarded cases. The associated variables for the confirmation of BSF in the multiple logistic model using a confidence interval (CI) of 95% were age (OR = 1.025 CI: 1.015-1.035), the presence of Amblyomma sculptum in the environment (OR = 1.629 CI: 1.097-2.439), the collection of ticks from horses (OR = 1.939 CI: 0.999-3.764), the presence of capybaras (OR = 1.467 CI: 1.009-2.138), an urban environment (OR = 1.515 CI: 1.036-2.231), and the existence of a dirty pasture (OR = 1.759 CI: 1.028-3.003). CONCLUSIONS : The factors associated with the confirmation of BSF cases included an urban environment, age, presence of the A. sculptum vector, the collection of ticks from horses, the presence of a capybara population, and a dirty pasture environment.