First report of canine rangeliosis in domestic dogs from different regions of Santa Catarina State, Brazil.

Rangeliosis is the disease caused by Rangelia vitalii, a parasite reported in dogs from southeastern and southern Brazil, Uruguay, Paraguay, and Argentina. This protozoan is transmitted by the ixodid Amblyomma aureolatum, and infects erythrocytes, leukocytes, and vascular endothelial cells of the host. The common clinical signs, such as prostration, fever, anemia, thrombocytopenia, anorexia, weight loss, and dehydration, are also found in other infections, like canine babesiosis and ehrlichiosis. The similar clinical presentation with other diseases, as well as the indistinguishable morphology with intraerythrocytic Babesia canis, complicates the disease diagnostic. In the present study, blood samples from dogs presenting clinical signs compatible with hemoparasitosis were investigated for rangeliosis. The dogs were treated at veterinary clinics in the cities of Blumenau and Lages, in the State of Santa Catarina, Brazil. Blood samples from 17 dogs were analyzed by PCR. The samples were screened by a conventional piroplasma-PCR and the positives confirmed by a specific R. vitalii-qPCR. Two animals (2/17; 11.8%) were positive for R. vitalii, one from Blumenau and the other from Lages. Both animals presented unspecific signs of hemoparasitosis, such as apathy, anemia, and anorexia. The results indicate the necessity of molecular assays for the proper identification of the hemoparasite, and to investigate the real prevalence of rangeliosis in the State of Santa Catarina.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil / Detecção molecular de Rangelia vitalii em cães do Estado do Paraná, Sul do Brasil

Abstract Rangelia vitalii infects erythrocytes, leukocytes and endothelial cells of dogs. The present study aimed to report the molecular detection confirmed by sequencing of R. vitalii in the state of Paraná, as well as describe the clinical, hematological and biochemical alterations of the infected dogs. Three sick dogs from the metropolitan area of Curitiba, PR, Brazil, underwent a physical exam, and laboratory tests included hematology, biochemistry, polymerase chain reaction (PCR), and gene sequencing. Clinical signs included apathy, anorexia, and hemorrhage. Intra-erythrocytic and extracellular piroplasms were found on peripheral blood smears from all three dogs. Blood samples from these animals were positive for Babesia sp. by PCR targeting 18S rRNA. PCR products from all three dogs were sequenced, and BLAST analysis showed that the PCR-generated sequences were highly homologous with those of R. vitalii previously reported. Hematologic findings included severe anemia, shift of neutrophils to the regenerative left, and thrombocytopenia. Serum urea levels were increased in all three dogs, and direct bilirubin levels were elevated in one dog.

Resumo Rangelia vitalii infecta eritrócitos, leucócitos e células endoteliais de cães. O presente estudo objetivou relatar a detecção molecular confirmada por sequenciamento de R. vitalii no estado do Paraná e descrever as alterações clínicas, hematológicas e bioquímicas dos cães infectados. Três cães doentes da região metropolitana de Curitiba, PR, Brasil, foram submetidos a exame físico e exames laboratoriais que incluíram hematologia, bioquímica, reação em cadeia da polimerase (PCR) e sequenciamento genético. Os sinais clínicos incluíram apatia, anorexia e hemorragia. Piroplasmas intra-eritrocíticos e extracelulares foram encontrados em esfregaços de sangue periférico dos três cães. As amostras de sangue destes animais foram positivas para Babesia sp. pela PCR baseada no gene 18S rRNA. Os produtos de PCR dos três cães foram sequenciados e a análise de BLAST mostrou que as seqüências geradas por PCR eram altamente homólogas com as de R. vitalii previamente relatadas. Os achados hematológicos incluíram anemia grave, desvio de neutrófilos à esquerda regenerativo e trombocitopenia. Os níveis de uréia no soro aumentaram nos três cães, e os níveis de bilirrubina direta foram elevados em um cão.

Detection molecular of Rangelia vitalii in dogs from Parana State, Southern Brazil

Abstract Rangelia vitalii infects erythrocytes, leukocytes and endothelial cells of dogs. The present study aimed to report the molecular detection confirmed by sequencing of R. vitalii in the state of Paraná, as well as describe the clinical, hematological and biochemical alterations of the infected dogs. Three sick dogs from the metropolitan area of Curitiba, PR, Brazil, underwent a physical exam, and laboratory tests included hematology, biochemistry, polymerase chain reaction (PCR), and gene sequencing. Clinical signs included apathy, anorexia, and hemorrhage. Intra-erythrocytic and extracellular piroplasms were found on peripheral blood smears from all three dogs. Blood samples from these animals were positive for Babesia sp. by PCR targeting 18S rRNA. PCR products from all three dogs were sequenced, and BLAST analysis showed that the PCR-generated sequences were highly homologous with those of R. vitalii previously reported. Hematologic findings included severe anemia, shift of neutrophils to the regenerative left, and thrombocytopenia. Serum urea levels were increased in all three dogs, and direct bilirubin levels were elevated in one dog.

Resumo Rangelia vitalii infecta eritrócitos, leucócitos e células endoteliais de cães. O presente estudo objetivou relatar a detecção molecular confirmada por sequenciamento de R. vitalii no estado do Paraná e descrever as alterações clínicas, hematológicas e bioquímicas dos cães infectados. Três cães doentes da região metropolitana de Curitiba, PR, Brasil, foram submetidos a exame físico e exames laboratoriais que incluíram hematologia, bioquímica, reação em cadeia da polimerase (PCR) e sequenciamento genético. Os sinais clínicos incluíram apatia, anorexia e hemorragia. Piroplasmas intra-eritrocíticos e extracelulares foram encontrados em esfregaços de sangue periférico dos três cães. As amostras de sangue destes animais foram positivas para Babesia sp. pela PCR baseada no gene 18S rRNA. Os produtos de PCR dos três cães foram sequenciados e a análise de BLAST mostrou que as seqüências geradas por PCR eram altamente homólogas com as de R. vitalii previamente relatadas. Os achados hematológicos incluíram anemia grave, desvio de neutrófilos à esquerda regenerativo e trombocitopenia. Os níveis de uréia no soro aumentaram nos três cães, e os níveis de bilirrubina direta foram elevados em um cão.

Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil / Rangelia vitalii, Babesia spp. e Ehrlichia spp. em cães de Passo Fundo, estado do Rio Grande do Sul, Brasil

Abstract Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) and Ehrlichia spp. (gene dsb). Five (8.6%) of the 58 dogs were serologically positive for Babesia spp. and three (5.1%) for E. canis. Four dogs (6.8%) were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed.

Resumo Patógenos transmitidos por carrapatos são um problema emergente em todo o mundo, o trabalho objetivou diagnosticar os agentes causais da infecção em cães com suspeita de hemoparasitoses. Cinquenta e oito caninos com sinais clínicos como depressão, diáteses hemorrágicas e febre foram avaliados quanto à apresentação clínica, hemograma, esfregaço sanguíneo, sorologia pelo método de Imunofluorescência Indireta para os agentes Babesia vogeli e Ehrlichia canis e na PCR convencional para Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) e Ehrlichia spp. (gene dsb). Cinco (8,6%) dos 58 cães apresentaram sorologia positiva para Babesia spp. e três (5,1%) para E. canis. Quatro (6,8%) animais mostraram-se positivos para R. vitalii no diagnóstico molecular. Os produtos da PCR foram sequenciados e o DNA encontrado de R. vitalii mostrou 99% de identidade genética com amostras de R. vitalii isoladas no Brasil. Não foi observada a presença de Babesia spp. e E. canis na PCR dos cães avaliados. Os resultados indicaram a presença de R. vitalii e exposição a Babesia spp. e Ehrlichia spp. entre os cães analisados.

First report of Rangelia vitalii infection (canine rangeliosis) in Argentina.

A 12-year old mixed breed neutered bitch from Misiones, Argentina, was presented with a history of fever and epistaxis. Blood, bone marrow, and lymph node samples were collected for hematology and cytology. Mild regenerative anemia was recorded and large, round, poorly stained piroplasms (>2.5 µm) were found within erythrocytes in blood and lymph node smears. Nested PCR-RFLP on blood and bone marrow samples was positive for piroplasm DNA. The 18S rRNA gene of piroplasms was targeted. A restriction pattern of a previously unreported piroplasm was observed. The PCR product was sequenced, and the sequence obtained had 99% identity with the Rangelia vitalii sequences from Brazil when compared by BLAST analysis. Further characterization of the detected piroplasm consisted of nearly full-length sequencing (1668 bp) of the 18S rRNA gene of this organism. Those sequences were deposited in GenBank. A phylogenetic analysis indicated that they clustered together with R. vitalii from Brazil but separately from large Babesia species of dogs such as Babesia canis, and from species of Theileria of dogs as well. This is the first report of R. vitalii infection in Argentina, and the first case of canine rangeliosis diagnosed outside Brazil.