A new focus of spotted fever caused by Rickettsia parkeri in Brazil

ABSTRACT Spotted fever (SF) is a tick-borne zoonosis caused by bacteria of the genus Rickettsia . The disease varies in severity from mild clinical forms to severe cases. In Brazil, Rickettsia rickettsii SF is the most serious rickettsiosis and can result in death if not diagnosed and treated at the onset of symptoms. The SF mild form is caused by Rickettsia parkeri strain Atlantic Rainforest, and this etiological agent has been reported in the South, Southeast and Northeast regions of the country, in areas of preserved or little antropized Atlantic Rainforest. Amblyomma ovale is the proven vector and dogs are the hosts associated with the bioagent cycle. During a SF case investigation in Paraty municipality, Rio de Janeiro State, an Atlantic Rainforest biome area in Southeastern Brazil, the human pathogen R. parkeri strain Atlantic Rainforest was detected by PCR in a sample of human skin inoculation eschar and in a female A. ovale tick collected from a dog. These results expand the known area of occurrence of this mild form rickettsiosis in Brazil. In addition, the results of the present study indicate the importance of implementing programs to control canine ectoparasites and to raise awareness of the risks of infection, signs and symptoms of SF caused by R. parkeri strain Atlantic Rainforest.

First molecular detection of Rickettsia sp. strain Atlantic rainforest in Amblyomma ovale ticks from Espírito Santo state, Brazil / Primeira detecção molecular da Rickettsia sp. cepa Mata Atlântica em carrapatos Amblyomma ovale do estado do Espírito Santo, Brasil

Abstract Espírito Santo state (southeastern Brazil) is considered an endemic area for spotted fever group rickettsioses. In February 2017, we received in our laboratory seven unfed Amblyomma ovale adult ticks collected by a farmer from his clothes and body (not attached) during a working day in the rural area of Ibiraçu municipality, Espírito Santo state. By polymerase chain reaction (PCR) analyses, targeting gltA and ompA rickettsial genes, the DNA of Rickettsia was detected in 6/7 (85.7%) A. ovale. In all cases, DNA sequencing of PCR products revealed that consensus sequences of both genes were 100% identical to gltA and ompA corresponding sequences of Rickettsia sp. strain Atlantic rainforest retrieved from GenBank. This study reports the first molecular detection of Rickettsia sp. strain Atlantic rainforest in A. ovale ticks from Espírito Santo state. Our findings indicate a new Brazilian state in the southeast region at risk of human infection with this tick-borne emerging rickettsial agent.

Resumo O estado do Espírito Santo (Sudeste do Brasil) é considerado área endêmica para riquetsioses do Grupo Febre Maculosa. Em fevereiro de 2017, recebemos em nosso laboratório sete carrapatos adultos Amblyomma ovale não ingurgitados, coletados por um fazendeiro nas suas roupas e corpo (não fixadas) durante um dia de trabalho, em área rural do municipio de Ibiraçu, estado do Espírito Santo. Por meio de reação em cadeia da polimerase (PCR), amplificando os genes riquetsiais gltA e ompA , foi detectado ADN de Rickettsia em 6/7 (85,7%) dos A. ovale . O sequenciamento dos produtos de PCR indicou que as sequências consenso de ambos genes foram 100% idênticos às sequências correspondentes dos genes gltA e ompA da Rickettsia sp. cepa Mata Atlântica recuperadas do GenBank. Este estudo relata a primeira detecção molecular da Rickettsia sp. cepa Mata Atlântica em carrapatos A. ovale do estado do Espírito Santo. Nossos resultados apontam um novo estado brasileiro da região Sudeste com risco de infecção humana por este agente rickettsial emergente transmitido por carrapatos.

Ixodidae fauna of domestic dogs in Parana, southern Brazil / Fauna ixodídica em cães domésticos no Paraná, Sul do Brasil

Abstract The present study aimed to contribute towards identification and registration of tick species that parasitize dogs in rural and urban areas of three mesoregions of Paraná, southern Brazil, and to estimate the rate of occurrence of each species. Fifty-six dogs with ticks living in three mesoregions: Metropolitana de Curitiba (MC), Centro Oriental (COP) and Centro Sul Paranaense (CSP), were used in the study. From these 56 dogs, 253 ticks were collected and were identified and morphologically characterized according to the species. Among all the ticks, 69.6% were identified as belonging to the species Rhipicephalus sanguineus sensu lato (s. l.); 28.1% as Amblyomma aureolatum and 2.4% as Amblyomma ovale. Among the dogs in MC that were evaluated, 57.7% were parasitized by R. sanguineus s. l., 38.5% by A. aureolatum and 3.8% by A. ovale; while in COP, 72.4% of the dogs were parasitized by A. aureolatum and 27.6% by R. sanguineus s. l.. In CSP, one tick was obtained, which was identified as A. aureolatum.

Resumo O presente estudo objetivou contribuir com a identificação e o registro das espécies de carrapatos que parasitam cães de áreas rurais e urbanas de três mesorregiões do Paraná, Sul do Brasil, e estimar a taxa de ocorrência de cada espécie. Cinquenta e seis cães com carrapatos, provenientes das mesorregiões: Metropolitana de Curitiba (MC), Centro Oriental (COP) e Centro Sul Paranaense (CSP) foram utilizados no estudo. Dos 56 cães, foram coletados 253 carrapatos que foram identificados e caracterizados morfologicamente de acordo com a espécie. Do total de carrapatos, 69,6% foram identificadas como pertencentes à espécie Rhipicephalus sanguineus s. l.; 28,1% como Amblyomma aureolatum e 2,4% como Amblyomma ovale. Dentre os animais avaliados, provenientes da MC, 57,7% estavam parasitados por R. sanguineus s. l., 38,5% por A. aureolatum e 3,8% por A. ovale; enquanto na COP 72,4% dos cães foram parasitados por A. aureolatum e 27,6% por R. sanguineus s. l.. Na CSP foi obtido um carrapato, identificado como A. aureolatum.

Ticks infesting wild small rodents in three areas of the state of São Paulo, Brazil / Carrapatos infestando pequenos roedores silvestres em três municípios do estado de São Paulo, Brasil

ABSTRACT: From May to September 2011, a total of 138 wild rodents of the Cricetidae family were collected in the cities of Anhembi, Bofete and Torre de Pedra, in São Paulo State. All animals were examined for the presence of ticks, which were collected and identified at species level in the laboratory by morphological examination (for adults, nymphs and larvae) and molecular biology, by sequencing of a fragment of the 16S mitochondrial rDNA (for larvae). A total of 47 ticks (21 larvae and 26 nymphs) were collected from rodents, identified as 21 larvae and 23 nymphs of Amblyomma ovale (collected on Oligoryzomys nigripes, Oligoryzomys flavescens, Sooretamys agouya and Nectomys squamipes), two nymphs of Ixodes schulzei (O. nigripes and O. flavescens) and an Amblyomma sculptum nymph in S. agouya. The overall prevalence of parasitism by A. ovale was 4.3% for larvae, and 10.1% for nymphs. The mean intensity of parasitism was 3.5 larvae/parasitized animal, and 1.6 nymphs/parasitized animal. One O. nigripes was found parasitized by both larva and nymph of A. ovale, and another O. nigripes was parasitized simultaneously by an I. schulzei nymph and five A. ovale nymphs. This research reports the following unpublished records: A. ovale on O. flavescens, O. nigripes and S. agouya; A. sculptum on S. agouya; and I. schulzei on O. flavescens and O. nigripes.

RESUMO: De maio a setembro de 2011, um total de 138 roedores silvestres da família Cricetidae foram coletados nos municípios de Anhembi, Bofete e Torre de Pedra, no estado de São Paulo. Todos os animais foram examinados quanto à presença de carrapatos, os quais foram coletados e identificados ao nível de espécie em laboratório, através de análises morfológicas (para adultos, ninfas e larvas) e por biologia molecular para confirmar estas análises, através do sequenciamento de um fragmento do gene 16S rDNA mitocondrial (para uma parte das larvas). Um total de 47 carrapatos (21 larvas e 26 ninfas) foi coletado dos roedores, sendo identificados como 21 larvas e 23 ninfas de Amblyomma ovale (coletados sobre Oligoryzomys nigripes, Oligoryzomys flavescens, Sooretamys agouya e Nectomys squamipes), duas ninfas de Ixodes schulzei (em O. nigripes e O. flavescens) e uma ninfa de Amblyomma sculptum em S. agouya. A prevalência geral de parasitismo por A. ovale foi de 4,3% e de 10,1% para larvas e ninfas, respectivamente. As intensidades médias de parasitismo foram de 3,5 larvas/animal parasitado e de 1,6 ninfas/animal parasitado. Um O. nigripes foi encontrado parasitado simultaneamente por larva e ninfa de A. ovale, e outro O. nigripes estava parasitado simultaneamente por uma ninfa de I. schulzei e cinco ninfas de A. ovale. O presente trabalho reporta os seguintes registros inéditos: A. ovale em O. flavescens, O. nigripes e S. agouya; A. sculptum em S. agouya; e I. schulzei em O. flavescens e O. nigripes.

Ticks and fleas in crab-eating fox (Cerdocyon thous) of Pernambuco state, Brazil / Carrapatos e pulgas em cachorro-do-mato (Cerdocyon thous) do Estado de Pernambuco, Brasil

This study aimed to identify the species of fleas and ticks of Cerdocyon thous from the state of Pernambuco. Animals (n = 20) were examined, with 30% (6/20) ectoparasitized. Fleas (n = 16) and ticks (n = 17) parasitizing free-living crab-eating fox and captive in state of Pernambuco were collected. The fleas were identified as: Pulex irritans and Ctenocephalides felis; and the ticks were: Rhipicephalus sanguineus sensu lato and Amblyomma ovale. The presence of ectoparasites of domestic animals parasitizing C. thous, suggests a close contact of this species to the peridomicile. Furthermore, this is the first report of the occurrence of A. ovale in C. thous in northeastern Brazil.(AU)

Objetivou-se identificar as espécies de pulgas e carrapatos de Cerdocyon thous provenientes do estado Pernambuco. Foram examinados 20 animais, estando 30% (6/20) ectoparasitados. Foram coletadas 16 pulgas e 17 carrapatos em cachorros-do-mato de vida livre e de cativeiro no estado de Pernambuco. As pulgas foram identificadas como: Pulex irritans e Ctenocephalides felis; e os carrapatos foram: Rhipicephalus sanguineus sensu lato e Amblyomma ovale. A presença de ectoparasitos de animais domésticos parasitando C. thous, sugere a aproximação de indivíduos desta espécie ao peridomicílio. Além disso, este é o primeiro relato da ocorrência de A. ovale em C. thous no nordeste brasileiro.(AU)

Parasitismo por Amblyomma ovale e Amblyomma fuscum (Acari: Ixodidae) em Dasypus novemcinctus (Xenarthra: Dasypodidae) no Brasil / Parasitism by Amblyomma ovale and Amblyomma fuscum (Acari: Ixodidae) on Dasypus novemcinctus (Xenarthra: Dasypodidae) in Brazil

Carrapatos são ectoparasitos hematófagos importantes para a saúde pública e animal por transmitirem agentes infecciosos durante a hematofagia, bem como causarem injúrias a seus hospedeiros. A ocorrência dos carrapatos Amblyomma ovale e Amblyomma fuscum é registrada, pela primeira vez, para o Brasil, parasitando tatu-galinha (Dasypus novemcinctus).(AU)

Ticks are important bloodsucking ectoparasites for public and animal health for transmitting infectious agents during blood feeding and causing injuries to their hosts. The authors report the occurrence of Amblyomma ovale and Amblyomma fuscum for the first time in Brazil, parasitizing the nine-banded armadillo (Dasypus novemcinctus).(AU)

Diagnóstico de hepatozoon spp. en perros (Canis familiaris) y sus vectores en áreas rurales de los Estados Lara y Yaracuy-Venezuela / Diagnosis of hepatozoon spp. in dogs (Canis familiaris) and its vectors from rural areas of Lara and Yaracuy States, Venezuela

El objetivo de este trabajo fue diagnosticar Hepatozoon spp. en perros de áreas rurales en algunos municipios de los estados Lara y Yaracuy, así como su relación con los potenciales artrópodos vectores involucrados en su transmisión. Babesia canis vogeli, Ehrlichia canis y en menor frecuencia Hepatozoon canis son los agentes parasitarios más importantes en caninos teniendo todos ellos como vector a Rhipicephalus sanguineus. Estos hemotrópicos infectan frecuentemente a los animales sin causarles sintomatología clínica y en otros casos les causan enfermedad de gravedad variable. La asociación de estos agentes es muy común, lo que favorece la gravedad de los síntomas clínicos. Además de R. sanguineus, otros géneros de garrapatas pueden ser encontradas parasitando perros, como Amblyomma e Ixodes. Durante el estudio se realizaron frotis sanguíneos de sangre periférica de la oreja de 300 perros y se colectaron 604 garrapatas por medio de extracción manual. Estos ixódidos fueron identificados taxonómicamente en el Laboratorio de Parasitología con la ayuda de claves dicotómicas y luego fueron sometidas a disección en busca de fases evolutivas de Hepatozoon spp. Los resultados fueron analizados a través de estadísticas no paramétricas indicando la presencia o ausencia de Hepatozoon spp. y la identificación de los vectores involucrados. Se observó Hepatozoon spp. en las muestras de sangre periférica de 11 de los perros estudiados, correspondiendo a tres perros del estado Lara y ocho al estado Yaracuy. En relación a los vectores, se identificaron en el estado Lara, 557 Rhipicephalus sanguineus, 11 Amblyomma ovale y 10 Ixodes spp. y en el estado Yaracuy solamente se diagnosticaron 26 especímenes de Amblyomma ovale. Es importante destacar la presencia de Hepatozoon spp. en áreas rurales donde no se identificó R. sanguineus.

The purpose of this research was to diagnose Hepatozoon spp. in dogs from rural areas in some municipalities of the States of Lara and Yaracuy, Venezuela, and its relationship with potential arthropods vectors involved in its transmission. Babesia canis vogeli, Ehrlichia canis, and less frequently, Hepatozoon canis, are the most important hemoparasites of dogs, all having as a common vector Rhipicephalus sanguineus. These hemoparasites frequently infest both domestic and rural animals without overt clinical symptoms; in other cases, they cause diseases of varied severity. Coinfections by hemoparasites are very common, which favors the severity of clinical symptoms. Besides R. sanguineus, other tick genera like Amblyomma and Ixodes, can be found infesting dogs. To carry out this study, thin blood smears from peripheral ear veins of 300 dogs were performed; and 604 ticks were collected by manual extraction. These ixodids were taxonomically identified at the Parasitology Laboratory, with the help of dichotomous taxonomic keys. Then, they were subjected to dissections looking for evolutionary stages of Hepatozoon spp. Results obtained were analyzed through non-parametric statistics, showing the presence or absence of Hepatozoon spp. and the identification of the involved vectors. Hepatozoon spp. was found in samples of peripheral blood in 11 of the dogs studied, three from the State of Lara and eight from the State of Yaracuy. Regarding vectors, 557 Rhipicephalus sanguineus, 11 Amblyomma ovale and 10 Ixodes spp. were identified in Lara, whereas only 26 specimens of Amblyomma ovale were identified in Yaracuy. It is important to underscore the presence of Hepatozoon spp. in rural areas where R. sanguineus was not identified.

Ocorrência de Amblyomma aureolatum (Pallas, 1772) e A. ovale (Kock, 1844) (Acari: Ixodidae) parasitando Alouatta clamitans Cabrera, 1940 (Primates: Atelidae) na região norte do estado de Santa Catarina / Ocurrence of Amblyomma aureolatum (Pallas, 1772) and A. ovale (Kock, 1844) (Acari: Ixodidae) parasitizing Alouatta clamitans Cabrera, 1940 (Primates: Atelidae) in the north of Santa Catarina state, Brazil

The authors confirm the presence of one female and two males of Amblyomma aureolatum and five females and one male of A. ovale parasitizing southern brown howler monkeys in the municipalities of Indaial, Blumenau, Garuva, and Jaraguá do Sul, Santa Catarina State, Brazil.

Especies de Amblyomma (acari: ixodidae) parasitando perros (canis familiaris) en áreas rurales de los estados Lara, Yaracuy, Carabobo y Falcón Venezuela / Species of Amblyomma (acari: ixodidae) parasitizing dogs (canis familiaris) from rural areas of Lara, Yaracuy, Carabobo y Falcón Venezuela

Un total de 50 caninos de áreas rurales de los estados Lara, Yaracuy, Carabobo y Falcón, Venezuela, fueron examinados visualmente y mediante palpación cutánea con el fin de detectar la presencia de garrapatas. Ninguno de los animales se encontró parasitado por Rhipicephalus sanguineus, la garrapata común del perro. Se colectaron 139 especimenes adultos del género Amblyomma Koch, 1844: 72 hembras y 65 machos de la especie Amblyomma ovale Koch, 1844, y dos hembras de la especie Amblyomma triste Koch, 1844. La identificación de los especimenes se realizó mediante observación en lupa estereoscópica y con la ayuda de claves taxonómicas. Los Estados donde se colectó el mayor número de ixódidos fueron Falcón con 52 especimenes y Lara con 49, representando 38 y 36%, respectivamente. A. ovale ha sido reportada como vector natural de Hepatozoon canis en áreas rurales. La especie A. triste fue colectada sólo en un canino del estado Yaracuy. Esta especie ha sido incriminada como el vector biológico de varios agentes rickettsiales patógenos, tanto para humanos como para animales domésticos y salvajes, por lo que se resalta la importancia del género Amblyomma en la transmisión de enfermedades zoonóticas.

Fifty dogs from the rural areas of Lara, Yaracuy, Carabobo and Falcón States of Venezuela were sampled through the visual and touch techniques in search of the presence of ticks. All the dogs were free of the brown dog tick Rhipicephalus sanguineus. A total of 139 adult ticks of the genus Amblyomma Koch, 1844 were collected: of them, 65 males and 72 females were identified as Amblyomma ovale Koch, 1844, whereas two females belonged to the species Amblyomma triste Koch, 1844. The observation through stereomicroscope, as well as the use of taxonomic keys, were employed in the identification of the collected specimens. Most of the ticks were collected in Falcón and Lara States, totalling 52 (38%) and 49 (36%), respectively. The species A. ovale has been reported as a natural vector of the protozoan Hepatozoon canis in rural areas. A. triste was collected only in one dog in Yaracuy State. This species has been incriminated as a biological vector of several rickettsial agents to the men and animals. The actual role of the genus Amblyomma as vector of zoonotic disease agents is highlighted.

Aislamiento de Rickettsia bellii a partir de Garrapatasamblyomma ovale y Amblyomma incisum procedentes del sur de Brasil / Isolation of Rickettsia bellii from Amblyomma ovale and Amblyomma incisum ticks from southern Brazil

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 stateof São Paulo, Brazil. Each tick was tested using the hemolymph assay; samples from positiveticks were placed in shell vials in order to isolate rickettsiae and subsequently grown in Verocells. Amplification of three rickettsial genes (gltA, htrA and ompA) was attempted usingpolymerase chain reaction (PCR) for each isolate obtained. Amplicons were subsequentlysequenced. 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 successfullyisolated from these ticks; however establishment in Vero cell culture was successful only forthe isolate from A. ovale. Bacterial contamination in the first cell passage of the A. incisumisolate precluded successful isolation of the organism. PCR products were obtained with thegltA and htrA primers for the two isolates, but no product was obtained with the ompAprimers. 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 thefirst report of R. bellii infecting A. incisum and the first successful isolation from A. ovale.