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.Keywords: Hemoparasites, piroplasms, sequencing, dogs, nambiuvú, indirect immunofluorescence. ResumoPatógenos 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.
Equine piroplasmosis is a disease caused by the hemoparasites Babesia caballi and Theileria equi and is considered to be the most important parasitic infection affecting Equidae. The objective of the present study was to carry out an epidemiological molecular and serological survey for the presence of these two protozoal organisms in equids from the northwestern region of the State of Rio Grande do Sul (RS), south Brazil. For this purpose, blood samples were collected from 90 equids in the city of Passo Fundo, RS, Brazil. Those were animals used for sport activities, outdoor recreational riding, and work including cattle herding and mounted patrol. Anti-T. equi and anti-B. caballi IgG antibodies were detected in the sera of those animals by commercial ELISA kits. The molecular diagnosis of equine piroplasmosis due to T. equi or B. caballi (or both) consisted in the amplification of the 18S rRNA gene by nested PCR followed by sequencing of the amplified PCR product and sequence comparison and phylogenetic analysis of the isolates; 17 (18.9%) and 5 (5.55%) out of the 90 serum samples tested in this study were positive for T. equi and B. caballi, respectively. Piroplasmid 18S rRNA gene fragments were detected by PCR in 24.4% (22/90) of the samples analysed and shared 99-100% identity with sequences of T. equi by BLASTn. Samples for the phylogenetic analysis were divided into 2 groups. In group A, there was close phylogenetic relationship between 4 sequences and sequences previously reported along the US-Mexico border, in South Africa, and in Brazil. There was a phylogenetic proximity between 5 samples from group B and samples tested by other authors in the US and Spain. Variation of the 18S rRNA gene allowed the identification of 9 new T. equi genotypes in the geographical region studied.
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