Linfadenopatía transmitida por garrapata. Un caso pediátrico de rickettsiosis en atención primaria / Lymphadenopathy transmitted by tick. A pediatric case of rickettsiosis in primary care

La entidad llamada tick-borne lymphadenopathy, también conocida como dermacentor-borne necrosis eritema lymphadenopathy, se incluye dentro del diagnóstico diferencial de enfermedades transmitidas por garrapatas, que han aumentado considerablemente en los últimos años debido a una mayor observación clínica-epidemiológica y mejoría diagnóstica. Es una zoonosis causada por Rickettsia slovaca y transmitida por la picadura de la garrapata Dermacentor marginatus. Los pacientes afectados tienen una escara necrótica rodeada de un halo eritematoso en el cuero cabelludo y adenopatías regionales dolorosas. Se presenta a un varón de siete años que mostró, tras la extracción de una garrapata, una escara necrótica en el cuero cabelludo, con adenopatías y febrícula. La reacción en cadena de la polimerasa y el cultivo de la lesión confirmaron la presencia de Rickettsia slovaca. El paciente recibió azitromicina oral con buena respuesta. En Atención Primaria, es importante el seguimiento de las picaduras de garrapatas para detectar posibles enfermedades transmitidas por estas.

The entity called tick-borne lymphadenopathy, also known as dermacentor-borne necrosis eritema lymphadenopathy, is included in the differential diagnosis of diseases transmitted by ticks, which have increased considerably in recent years due to greater clinical-epidemiological observation and diagnostic improvement. It is a zoonosis caused by Rickettsia slovaca and transmitted by the bite of the Dermacentor marginatus tick. Affected patients have a necrotic eschar surrounded by an erythematous halo in the scalp, as well as painful regional lymphadenopathy. Linfadenopatía transmitida por garrapata. Un caso pediátrico de rickettsiosis en atención primaria Lymphadenopathy transmitted by tick. A pediatric case of rickettsiosis in primary care We present a seven-year-old male who shows, after the extraction of a tick, a necrotic eschar on the scalp, accompanied by lymphadenopathy and low-grade fever. The study of polymerase chain reaction and culture of the lesion confirm the presence of Rickettsia slovaca. The patient received oral azithromycin with a good response. In Primary Care, it is important to monitor the tick bites, to detect possible diseases transmitted by them.

Serological survey of Rickettsia in equids from Vale do Paraíba, São Paulo, Brazil, and their tick identification and molecular investigation of Rickettsia / Levantamento sorológico de Rickettsias em equinos no Vale do Paraíba, São Paulo, Brasil. Identificação e pesquisa molecular de Rickettsias em carrapatos

Brazilian spotted fever is a serious and lethal illness for humans and is caused by the Rickettsia rickettsii bacteria. In the state of São Paulo/SP (Brazil), the etiological agent of this disease is transmitted by the Amblyomma sculptum tick. It was already shown that horses infected with this bacteria produce a strong immune response and could be important sentinels for the detection of the disease in a proper region. The present investigation performed a serological survey in horses from five farms of Vale do Paraíba, São Paulo state, Brazil, searching for antibodies against, Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Rickettsia rhipicephali, and Rickettsia bellii. In each farm, ticks were also collected that were taxonomically identified and examined by real-time PCR for Rickettsia spp DNA. Blood samples were collected from 206 horses, and 334 ticks were picked up from these animals from January to December 2017. Eighty ticks were A. sculptum and 254 Dermacentor nitens. Of the blood samples, 7.3% seroconverted to Rickettsia spp. Of these, 0.97% had a positive serological response to R. bellii. None of the 80 A. sculptum ticks were positive through real-time PCR for Rickettsia spp. Although there was no detection of ticks infected by Rickettsia spp in five farms of Paraíba Valley, the horses presented serological positive reactions against this agent. Thus, further large studies should be conducted in the area targeting hosts and vectors to generate data for control measures of the transmission of Brazilian spotted fever(AU)

A febre maculosa brasileira é uma doença grave e letal para seres humanos causada pela bactéria Rickettsia rickettsii. No estado de São Paulo, SP, Brasil, o agente etiológico desta enfermidade é transmitido pelo carrapato Amblyomma sculptum. Conforme descrito na literatura científica, os cavalos infectados com esta bactéria produzem uma forte resposta imune e podem ser importantes sentinelas para a detecção da doença. A presente investigação realizou um levantamento sorológico em cavalos de cinco fazendas do Vale do Paraíba, São Paulo, Brasil, à procura de anticorpos contra Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Rickettsia rhipicephali e Rickettsia bellii. Em cada fazenda, também foram coletados carrapatos identificados taxonomicamente e examinados por PCR em tempo real para o DNA de Rickettsia spp. Foram coletadas amostras de sangue de 206 cavalos e coletados 334 carrapatos desses animais entre os meses de janeiro e dezembro de 2017. Oitenta carrapatos foram identificados como A. sculptum e 254 Dermacentor nitens. Das amostras de sangue, 7,3% soroconverteram para Rickettsia spp., sendo que, 0,97% apresentaram soropositividade homóloga para R. bellii. Nenhum dos 80 carrapatos de A. sculptum foi positivo com o emprego de PCR em tempo real para Rickettsia spp. Embora não tenham sido detectados carrapatos infectados por Rickettsia spp em cinco fazendas do Vale do Paraíba, os animais apresentaram reações sorológicas positivas para este agente. Assim, outros estudos abrangentes deverão ser realizados na área investigando hospedeiros e vetores, gerando dados para medidas de controle da transmissão da febre maculosa brasileira.(AU)

Life Cycle of Dermacentor everestianus Hirst, 1926 (Acari: Ixodidae) under Laboratory Conditions

This study investigated the development characteristics of Dermacentor everestianus under laboratory conditions. The time taken for D. everestianus to complete the whole life cycle was 110.2 days on average, and the average developmental durations of larvae and nymphs were 17.1 days and 29.5 days, respectively. The summation of the prefeeding, feeding, and preoviposition periods of females was 17.8 days, and the oviposition and egg incubation lasted for 18.1 days and 27.7 days, respectively. A highly positive correlation was observed between the weight of engorged female and the number of egg mass laid (r=0.947). The reproductive efficiency index and the reproductive fitness index were 7.1 and 6.1, respectively.

A rare case of tick infestation of the eyelid: case report and literature review / Um caso raro de infestação de carrapatos da pálpebra: relato de caso e revisão da literatura

ABSTRACT This article presents a case of tick infestation of the lower eyelid by a previously unreported species. A 71-year-old male presented with a tick attached to the lower eyelid. The tick was identified morphologically, and then molecularly via polymerase chain reaction (PCR) and sequencing of its DNA. In addition, a review of the literature relevant to the genera of ticks associated with infestation of the human eye is provided. The tick, which was in the nymphal developmental stage, was first identified according to taxonomic keys as Dermacentor sp. For complete species identification, 16s rDNA gene PCR and sequencing were performed, which showed that the tick was D. marginatus. Systematizing tick species could assist physicians in determining the potential for transmission of tick-borne human diseases.

RESUMO Este artigo apresenta um caso de infestação por carrapatos da pálpebra inferior por uma espécie previamente não declarada. Um homem de 71 anos de idade apresentou-se com um carrapato grudado na pálpebra inferior. O carrapato foi identificado morfologicamente, e, em seguida, uma estrutura molecular através de reacção em cadeia da polimerase (PCR) e a sequenciação do seu DNA. Além disso, uma análise da literatura pertinente aos gêneros de carrapatos associados à infestação do olho humano é fornecido. O carrapato, que estava em fase de desenvolvimento das ninfas, foi identificado pela primeira vez de acordo com chaves taxonômicas com o Dermacentor sp. Para identificação de espécies completa, gene 16S rDNA PCR e sequenciamento foram realizadas, que mostrou que o carrapato foi D. marginatus. Sistematizando espécie de carrapato poderia ajudar os médicos a determinar o potencial de transmissão de doenças humanas transmitidas por carrapatos.

Screening and Identification of Antigenic Proteins from the Hard Tick Dermacentor silvarum (Acari: Ixodidae)

In order to explore tick proteins as potential targets for further developing vaccine against ticks, the total proteins of unfed female Dermacentor silvarum were screened with anti-D. silvarum serum produced from rabbits. The results of western blot showed that 3 antigenic proteins of about 100, 68, and 52 kDa were detected by polyclonal antibodies, which means that they probably have immunogenicity. Then, unfed female tick proteins were separated by 12% SDS-PAGE, and target proteins (100, 68, and 52 kDa) were cut and analyzed by LC-MS/MS, respectively. The comparative results of peptide sequences showed that they might be vitellogenin (Vg), heat shock protein 60 (Hsp60), and fructose-1, 6-bisphosphate aldolase (FBA), respectively. These data will lay the foundation for the further validation of antigenic proteins to prevent infestation and diseases transmitted by D. silvarum.

Gene Cloning, Expression and Immunogenicity of the Protective Antigen Subolesin in Dermacentor silvarum

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.

First record of Borrelia burgdorferi B31 strain in Dermacentor nitens ticks in the northern region of Parana (Brazil)

The aim of this study was to investigate the presence of DNA of Borrelia burgdorferi sensu lato (s.l.) in ticks that feed on horses used for animal traction in rural Jataizinho, Parana, Brazil. Between February and June 2008, a total of 224 ticks was collected of which 75% were identified as Dermacentor nitens and 25% as Amblyomma cajenense. To amplify B. burgdorferi s.l. DNA, the intergenic space region (ISR) between the 5S (rrf) 23S (rrl) rRNA genes was used as targets for nested-PCR. Two ticks of the D. nitens species were positive for B. burgdorferi s.l. Both species showed a fragment of 184 bp, but the sequencing revealed 99.9% homology with the B. burgdorferi sensu stricto (s.s.) strain B31. These results showed, for the first time, the presence of spirochete DNA infecting ticks that parasitize horses used for animal traction, in the rural municipality mentioned. In conclusion, this study opens up promising prospects for determining the infection rate of B. burgdorferi s.s. genospecies or other species in the equine population, as well as the impact of the infection rate on Lyme disease in the state of Parana.

Ticks on buffaloes (Bubalusbubalis) in the state of Rio de Janeiro, Brazil / Carrapatos em búfalos (Bubalusbubalis) do estado do Rio de Janeiro, Brasil

Some tick species have been described parasitizing buffaloes, but reports from Brazil are rare. This study aimed to ascertain the species of ticks that parasitize buffaloes in the state of Rio de Janeiro. Seventeen farm properties were visited between June 2008 and December 2009, and buffaloes were inspected in relation to infestation. The ticks were identified and a semi-structured questionnaire on parasitism and treatment against ticks was filled out through interviews with the owners. The species identified were Rhipicephalus (Boophilus) microplus, Dermacentornitens and Amblyommacajennense. It was noted that 82.3% (14/17) of the properties visited presented animals infested with ticks, although 76.5% (13/17) had been administering acaricide treatment. Buffaloes in the state of Rio de Janeiro are often parasitized by ticks, especially in the larval and nymph stages.

Algumas espécies de carrapatos foram descritas parasitando bubalinos, porém são raros os relatos no Brasil. O presente trabalho teve como objetivo conhecer as espécies de carrapatos que parasitam bubalinos no estado do Rio de Janeiro. Dezessete propriedades foram visitadas de junho de 2008 a dezembro de 2009 e os bubalinos foram inspecionados para verificar infestação. Os carrapatos foram identificados e um questionário semi estruturado sobre o parasitismo e o tratamento contra carrapatos foram preenchidos por meio de entrevista com os proprietários. Foram identificadas as espécies Rhipicephalus (Boophilus) microplus, Dermacentornitens e Amblyommacajennense. Foi observado que 82,3% (14/17) das propriedades visitadas apresentavam animais parasitados por carrapatos, embora 76,5% (13/17) realizassem tratamento acaricida. Búfalos do estado do Rio de Janeiro são frequentemente parasitados por carrapatos, principalmente nos estágios de larvas e ninfas.

Spotted fever group Rickettsia infecting ticks (Acari: Ixodidae) in the state of Santa Catarina, Brazil

During 2006-2008, a total of 260 adult ticks were collected from domestic and wild animals in different regions of the state of Santa Catarina (SC), Brazil, including areas where human cases of Brazilian spotted fever have been reported. Collected ticks belonging to nine species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Amblyomma longirostre, Amblyomma ovale, Amblyomma tigrinum, Dermacentor nitens, Rhipicephalus microplus and Rhipicephalus sanguineus) were tested by polymerase chain reaction (PCR) for rickettsial infection. Overall, eight (3.1 percent) ticks were found to be infected with Rickettsia species. After sequencing the PCR products, we determined that the sequences generated from three A. aureolatum, one A. ovale and one R. sanguineus from the municipality of Blumenau, one A. ovale from the municipality of Águas Mornas and one A. ovale from the municipality of Urussanga were identical to the corresponding partial rickettsial ompA gene sequence of Rickettsia parkeri strain Atlantic rainforest. The sequence generated from one A. longirostre from Blumenau was 100 percent identical to the corresponding partial rickettsial ompA gene sequence of Rickettsia amblyommii strain AL. Because R. parkeri strain Atlantic rainforest was recently shown to have caused two cases of human spotted fever in other states of Brazil, the role of this rickettsial agent as a possible etiological agent of spotted fever in SC is discussed.

Ocorrência de carrapatos (acari: ixodidae) parasitando equinos no município de Jataizinho, Paraná / Ocurrencia de garrapatas (acari: ixodidae) parasitando equinos en el municipio de Jataizinho, Paraná / Occurence of ticks (acari: ixodidae) parasiting equines in the municipality of Jataizinho, Paraná

Devido a suspeita de anticorpos contra Borrelia burgdorferi sensu lato (s.l) em moradores da área rural do município de Jataizinho (PR), foram coletados, entre fevereiro a junho de 2008, ixodídeos adultos parasitando equinos de 21 propriedades rurais deste município. Os 224 ixodídeos adultos de equinos foram identificados como Amblyomma cajennense (25%) e Dermacentor nitens (75%). Nas áreas de coleta desta pesquisa, o parasitismo humano por carrapatos era frequentemente relatado pelos moradores rurais. O estudo dos ixodídeos parasitando equinos em relação à doença de Lyme é um assunto pouco explorado e entendido no Brasil. No estado do Paraná (PR) é a primeira vez que a doença de Lyme está sendo pesquisada, o que ressalta a importância deste estudo nos ixodídeos capturados em equinos, para o entendimento da epidemiologia desta enfermidade transmitida pelo carrapato em outras regiões do Brasil.

Due to the possibility of antibodies against Borrelia burgdorferi sensu lato (sl) in the rural area of Jataizinho (PR), adult ixodids parasitizing horses 21 farms in this municipality were collected between February and June 2008. 224 adult ixodid horses were identified as Amblyomma cajennense (25%) and Dermacentor nitens (75%). In the collection areas of this research, human infestation by ticks are frequently reported by rural residents. The study of ixodid parasites of horses in relation to Lyme disease is a little explored and understood subject in Brazil. In the state of Paraná (PR) it is the first time that Lyme disease is being studied empahsizing the importance of this study in ixodid from horses to understand the epidemiology of this disease transmitted by ticks in other regions of Brazil.

Debido a sospecha de anticuerpos contra Borrelia burgdorferi sensu lato (s.l) en moradores del área rural del municipio de Jataizinho (PR), se colectó entre febrero a junio de 2008, ixodídeos adultos parasitando equinos de 21 propiedades rurales de este municipio. Los 224 ixodídeos adultos de equinos fueron identificados como Amblyomma cajennense (25%) y Dermacentor nitens (75%). En las áreas de coleta de esta investigación, el parasitismo humano por garrapatas era frecuentemente relatado por los moradores rurales. El estudio de los ixodídeos parasitando equinos en relación a la enfermedad de Lyme es un asunto poco explotado y entendido en Brasil. En el estado de Paraná es la primera vez que la enfermedad de Lyme está siendo investigada, lo que resalta la importancia de este estudio en los ixodídeos capturados en equinos, para el entendimiento de la epidemiología de esta enfermedad transmitida por garrapata en otras regiones de Brasil.