Patógenos transmitidos por vetores na reprodução canina / Vector-borne pathogens in canine reproduction

O artigo apresenta uma revisão sobre os efeitos reprodutivos da infecção pelos principais patógenos transmitidos por vetores que acometem cães, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. e Trypanosoma spp. São discutidas a patologia genital, fertilidade masculina e feminina, transmissão venérea, transmissão vertical, transmissão horizontal mãe-filho e sua relevância na epidemiologia das doenças.(AU)

The article presents a review of the reproductive effects of infection by main vector-borne pathogens of dogs, Leishmania spp., Babesia spp., Ehrlichia spp., Anaplasma spp. and Trypanosoma spp. Genital pathology, male and female fertility, venereal transmission, vertical transmission, horizontal mother-offspring transmission and their relevance to the epidemiology of the diseases are discussed.(AU)

Occurrence of tick-borne diseases in domestic dogs in Belém, Pará, Brazil / Ocorrência de doenças transmitidas por carrapatos em cães domésticos em Belém do Pará, Brasil

Tick-borne blood cell pathogens, which are challenging to diagnose, are primarily detected using molecular techniques. Therefore, this study aimed to detect the main infectious agents involved in 50 cases of suspected hemoparasitosis in dogs treated at the Veterinary Hospital Mário Dias Teixeira of the Federal Rural University of the Amazon. Hematological parameters were evaluated, and blood samples were subjected to polymerase chain reaction (PCR) assays for DNA amplification of the following species: Ehrlichia canis, Anaplasma platys, and Babesia canis. The PCR test results indicated that the most prevalent infectious agent was E. canis, present in 12% (6/50) infected animals, followed by A. platys and B. canis, present in 8% (4/50) and 2% (1/50) infected animals, respectively. Regarding hematological analysis, the most relevant changes were anemia, lymphopenia, thrombocytopenia, leukocytosis, and leukopenia. The availability of molecular techniques allows the management of the most appropriate treatment to infected animals in a rapid and specific way.(AU)

Patógenos de células sanguíneas transmitidos por carrapatos, cujo diagnóstico é desafiador, são detectados princi-palmente por meio de técnicas moleculares. Dessa forma, este estudo teve como objetivo detectar os principais agentes infec-ciosos envolvidos em 50 casos de suspeita de hemoparasitose em cães atendidos no Hospital Veterinário Mário Dias Teixeira da Universidade Federal Rural da Amazônia. Parâmetros hematológicos foram avaliados e amostras de sangue foram subme-tidas a ensaios de reação em cadeia da polimerase (PCR) para amplificação de DNA das seguintes espécies: Ehrlichia canis, Anaplasma platys e Babesia canis. Os resultados do teste de PCR indicaram que o agente infeccioso mais prevalente foi E. canis, presente em 12% (6/50) dos animais infectados, seguido por A. platys e B. canis, presente em 8% (4/50) e 2% (1/50) animais infectados, respectivamente. Em relação à análise hematológica, as alterações mais relevantes foram anemia, linfopenia, trom-bocitopenia, leucocitose e leucopenia. A disponibilidade de técnicas moleculares permite o manejo do tratamento mais ade-quado aos animais infectados de forma rápida e específica.(AU)

Occurrence of tick-borne diseases in domestic dogs in Belém, Pará, Brazil / Ocorrência de doenças transmitidas por carrapatos em cães domésticos em Belém do Pará, Brasil

Tick-borne blood cell pathogens, which are challenging to diagnose, are primarily detected using molecular techniques. Therefore, this study aimed to detect the main infectious agents involved in 50 cases of suspected hemoparasitosis in dogs treated at the Veterinary Hospital Mário Dias Teixeira of the Federal Rural University of the Amazon. Hematological parameters were evaluated, and blood samples were subjected to polymerase chain reaction (PCR) assays for DNA amplification of the following species: Ehrlichia canis, Anaplasma platys, and Babesia canis. The PCR test results indicated that the most prevalent infectious agent was E. canis, present in 12% (6/50) infected animals, followed by A. platys and B. canis, present in 8% (4/50) and 2% (1/50) infected animals, respectively. Regarding hematological analysis, the most relevant changes were anemia, lymphopenia, thrombocytopenia, leukocytosis, and leukopenia. The availability of molecular techniques allows the management of the most appropriate treatment to infected animals in a rapid and specific way.

Patógenos de células sanguíneas transmitidos por carrapatos, cujo diagnóstico é desafiador, são detectados princi-palmente por meio de técnicas moleculares. Dessa forma, este estudo teve como objetivo detectar os principais agentes infec-ciosos envolvidos em 50 casos de suspeita de hemoparasitose em cães atendidos no Hospital Veterinário Mário Dias Teixeira da Universidade Federal Rural da Amazônia. Parâmetros hematológicos foram avaliados e amostras de sangue foram subme-tidas a ensaios de reação em cadeia da polimerase (PCR) para amplificação de DNA das seguintes espécies: Ehrlichia canis, Anaplasma platys e Babesia canis. Os resultados do teste de PCR indicaram que o agente infeccioso mais prevalente foi E. canis, presente em 12% (6/50) dos animais infectados, seguido por A. platys e B. canis, presente em 8% (4/50) e 2% (1/50) animais infectados, respectivamente. Em relação à análise hematológica, as alterações mais relevantes foram anemia, linfopenia, trom-bocitopenia, leucocitose e leucopenia. A disponibilidade de técnicas moleculares permite o manejo do tratamento mais ade-quado aos animais infectados de forma rápida e específica.

Occurrence of Babesia vogeli, Mycoplasma spp., Ehrlichia canis and Anaplasma spp. in a hospital dog population of western Paraná / Ocorrência de Babesia vogeli, Mycoplasma spp., Ehrlichia canis e Anaplasma spp. em uma população hospitalar de cães do Oeste do Paraná

Ehrlichia canis, Anaplasma platys, and Babesia vogeli are frequently detected in the veterinary routine with varied pathogenesis that can lead to death, mainly in co-infections. Although canine hemotropic mycoplasmas are considered of low clinical importance, they have recently gained prominence in molecular research. The present study aimed to analyze two hospital populations of dogs, randomly selected from the year 2013 and 2015, from the municipalities of Toledo (n=68) and Cascavel (n=107). Direct examination of blood smears, serology, and PCR were used to detect the presence of E. canis, A. platys, B. vogeli and Mycoplasma sp. Direct blood smear examination was negative for the investigated agents in all samples. Serum immunochromatography (SensPERT ™, VetAll Laboratories, Korea) in 175 animals showed that only 4% (n = 7) had anti-E. canis antibodies, while 60% (n = 105) were positive for Anaplasma sp. All PCR samples were negative for E. canis, while 18.28% (n = 32) were positive for A. platys, 3.42% (n = 6) positive for Babesia vogeli, and 7.41% (n = 13) positive for Mycoplasma sp. The study revealed the presence of A. platys in Toledo and Cascavel by PCR, which accentuates the need to monitor vector populations and usual hosts and to evaluate the potential risk of infection in humans.

Ehrlichia canis, Anaplasma platys e Babesia vogeli são frequentemente detectados na rotina veterinária com patogênese variada que pode levar à morte, principalmente em coinfecções. Embora os micoplasmas hemotrópicos caninos sejam considerados de baixa importância clínica, recentemente ganharam destaque na pesquisa molecular. O presente estudo teve como objetivo analisar duas populações hospitalares de cães, selecionadas aleatoriamente no ano de 2013 e 2015, dos municípios de Toledo (n = 68) e Cascavel (n = 107). O exame direto de esfregaços de sangue, sorologia e PCR foram usados para detectar a presença de E. canis, A. platys, B. vogeli e Mycoplasma sp. O esfregaço sanguíneo direto foi negativo para os agentes investigados em todas as amostras. A imunocromatografia sérica (SensPERT ™, VetAll Laboratories, Coréia) em 175 animais mostrou que apenas 4% (n = 7) tinham anticorpos contra E.canis, enquanto 60% (n = 105) foram positivos para Anaplasma sp. Todas as amostras de PCR foram negativas para E. canis, enquanto 18,28% (n = 32) foram positivas para A. platys, 3,42% (n = 6) positivas para Babesia vogeli e 7,41% (n = 13) positivas para Mycoplasma sp. O estudo revelou a presença de A. platys em Toledo e Cascavel por PCR, o que acentua a necessidade de monitorar populações de vetores e hospedeiros usuais e avaliar o risco potencial de infecção em humanos.

Occurrence of Babesia vogeli, Mycoplasma spp., Ehrlichia canis and Anaplasma spp. in a hospital dog population of western Paraná / Ocorrência de Babesia vogeli, Mycoplasma spp., Ehrlichia canis e Anaplasma spp. em uma população hospitalar de cães do Oeste do Paraná

Ehrlichia canis, Anaplasma platys, and Babesia vogeli are frequently detected in the veterinary routine with varied pathogenesis that can lead to death, mainly in co-infections. Although canine hemotropic mycoplasmas are considered of low clinical importance, they have recently gained prominence in molecular research. The present study aimed to analyze two hospital populations of dogs, randomly selected from the year 2013 and 2015, from the municipalities of Toledo (n=68) and Cascavel (n=107). Direct examination of blood smears, serology, and PCR were used to detect the presence of E. canis, A. platys, B. vogeli and Mycoplasma sp. Direct blood smear examination was negative for the investigated agents in all samples. Serum immunochromatography (SensPERT ™, VetAll Laboratories, Korea) in 175 animals showed that only 4% (n = 7) had anti-E. canis antibodies, while 60% (n = 105) were positive for Anaplasma sp. All PCR samples were negative for E. canis, while 18.28% (n = 32) were positive for A. platys, 3.42% (n = 6) positive for Babesia vogeli, and 7.41% (n = 13) positive for Mycoplasma sp. The study revealed the presence of A. platys in Toledo and Cascavel by PCR, which accentuates the need to monitor vector populations and usual hosts and to evaluate the potential risk of infection in humans.(AU)

Ehrlichia canis, Anaplasma platys e Babesia vogeli são frequentemente detectados na rotina veterinária com patogênese variada que pode levar à morte, principalmente em coinfecções. Embora os micoplasmas hemotrópicos caninos sejam considerados de baixa importância clínica, recentemente ganharam destaque na pesquisa molecular. O presente estudo teve como objetivo analisar duas populações hospitalares de cães, selecionadas aleatoriamente no ano de 2013 e 2015, dos municípios de Toledo (n = 68) e Cascavel (n = 107). O exame direto de esfregaços de sangue, sorologia e PCR foram usados para detectar a presença de E. canis, A. platys, B. vogeli e Mycoplasma sp. O esfregaço sanguíneo direto foi negativo para os agentes investigados em todas as amostras. A imunocromatografia sérica (SensPERT ™, VetAll Laboratories, Coréia) em 175 animais mostrou que apenas 4% (n = 7) tinham anticorpos contra E.canis, enquanto 60% (n = 105) foram positivos para Anaplasma sp. Todas as amostras de PCR foram negativas para E. canis, enquanto 18,28% (n = 32) foram positivas para A. platys, 3,42% (n = 6) positivas para Babesia vogeli e 7,41% (n = 13) positivas para Mycoplasma sp. O estudo revelou a presença de A. platys em Toledo e Cascavel por PCR, o que acentua a necessidade de monitorar populações de vetores e hospedeiros usuais e avaliar o risco potencial de infecção em humanos.(AU)

Molecular, serological, and parasitological detection of Babesia vogeli in dogs in the state of Piauí, Brazil / Detecção molecular, sorológica e parasitológica de Babesia vogeli em cães do estado do Piauí, Brasil

Studies on canine babesiosis in northeastern Brazil are scarce, although the weather conditions in this region are favorable for the development of the tick vector. This study determined the prevalence of Babesia vogeli in dogs sampled in Teresina, state of Piauí, northeast Brazil, using direct and indirect diagnostic methods and performed a phylogenetic analysis of 18S rRNA sequences. A total of 315 dogs were screened during routine care regardless of clinical suspicion. Blood was collected by jugular venipuncture to perform indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR) and for parasite screening in peripheral blood smears. Positivity was 2.2% (7/315) by microscopy, 4.8% (15/315) by PCR, and 48.6% (153/315) by IFA. PCR amplified a 602-bp fragment of the piroplasmid 18S rRNA gene, and sequence alignment and analysis revealed 99% homology with B. vogeli isolates from other regions of Brazil and other countries. In addition, there was high variability among sequences from other northeast states of Brazil. This study is the first to perform the molecular analysis of B. vogeli in Piauí. The results demonstrate that canine babesiosis is endemic in dogs sampled in Teresina and that PCR may be the method of choice to perform parasite screening in this region.

Estudos sobre a babesiose canina são escassos no Nordeste do Brasil, apesar das condições climáticas favoráveis ao desenvolvimento do carrapato vetor. Esta pesquisa objetivou determinar a ocorrência de Babesia vogeli em cães amostrados em Teresina, estado do Piauí, região Meio Norte do Brasil, através de métodos diretos e indiretos de diagnóstico, além de realizar análise filogenética das sequências 18S rRNA de piroplasmídeos obtidas no estudo. Foram avaliados 315 cães atendidos em clínicas veterinárias, sob qualquer suspeita clínica. Desses animais, foi colhido sangue por venopunção jugular para Reação de Imunofluorescência Indireta (RIFI) e Reação em Cadeia pela Polimerase (PCR). Além disso, esfregaços de sangue periférico foram realizados para pesquisa direta do parasita. A positividade dos animais foi de 2,2% (7/315) ao esfregaço sanguíneo, 4,8% (15/315) à PCR e 48,6% (153/315) à RIFI. O sequenciamento de amostras positivas à PCR resultou em um fragmento de 602 pb do gene 18S rRNA de piroplasmídeos, cujo alinhamento e análise da sequência revelaram 99% de homologia com isolados de B. vogeli de outras regiões do Brasil, além de outros países. É interessante ressaltar que, comparando isolados em diferentes estados do Nordeste, a homologia pode ser bastante variável. Esses são os primeiros resultados sobre a análise molecular de B. vogeli no Estado do Piauí. Além disso, este estudo demonstra que a babesiose canina é endêmica em cães de Teresina, Nordeste do Brasil, e que a PCR pode ser o método de escolha para diagnóstico da doença nessas áreas.

Molecular, serological, and parasitological detection of Babesia vogeli in dogs in the state of Piauí, Brazil / Detecção molecular, sorológica e parasitológica de Babesia vogeli em cães do estado do Piauí, Brasil

Studies on canine babesiosis in northeastern Brazil are scarce, although the weather conditions in this region are favorable for the development of the tick vector. This study determined the prevalence of Babesia vogeli in dogs sampled in Teresina, state of Piauí, northeast Brazil, using direct and indirect diagnostic methods and performed a phylogenetic analysis of 18S rRNA sequences. A total of 315 dogs were screened during routine care regardless of clinical suspicion. Blood was collected by jugular venipuncture to perform indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR) and for parasite screening in peripheral blood smears. Positivity was 2.2% (7/315) by microscopy, 4.8% (15/315) by PCR, and 48.6% (153/315) by IFA. PCR amplified a 602-bp fragment of the piroplasmid 18S rRNA gene, and sequence alignment and analysis revealed 99% homology with B. vogeli isolates from other regions of Brazil and other countries. In addition, there was high variability among sequences from other northeast states of Brazil. This study is the first to perform the molecular analysis of B. vogeli in Piauí. The results demonstrate that canine babesiosis is endemic in dogs sampled in Teresina and that PCR may be the method of choice to perform parasite screening in this region.(AU)

Estudos sobre a babesiose canina são escassos no Nordeste do Brasil, apesar das condições climáticas favoráveis ao desenvolvimento do carrapato vetor. Esta pesquisa objetivou determinar a ocorrência de Babesia vogeli em cães amostrados em Teresina, estado do Piauí, região Meio Norte do Brasil, através de métodos diretos e indiretos de diagnóstico, além de realizar análise filogenética das sequências 18S rRNA de piroplasmídeos obtidas no estudo. Foram avaliados 315 cães atendidos em clínicas veterinárias, sob qualquer suspeita clínica. Desses animais, foi colhido sangue por venopunção jugular para Reação de Imunofluorescência Indireta (RIFI) e Reação em Cadeia pela Polimerase (PCR). Além disso, esfregaços de sangue periférico foram realizados para pesquisa direta do parasita. A positividade dos animais foi de 2,2% (7/315) ao esfregaço sanguíneo, 4,8% (15/315) à PCR e 48,6% (153/315) à RIFI. O sequenciamento de amostras positivas à PCR resultou em um fragmento de 602 pb do gene 18S rRNA de piroplasmídeos, cujo alinhamento e análise da sequência revelaram 99% de homologia com isolados de B. vogeli de outras regiões do Brasil, além de outros países. É interessante ressaltar que, comparando isolados em diferentes estados do Nordeste, a homologia pode ser bastante variável. Esses são os primeiros resultados sobre a análise molecular de B. vogeli no Estado do Piauí. Além disso, este estudo demonstra que a babesiose canina é endêmica em cães de Teresina, Nordeste do Brasil, e que a PCR pode ser o método de escolha para diagnóstico da doença nessas áreas.(AU)

Ticks and Tick-Borne Pathogens of the Caribbean: Current Understanding and Future Directions for More Comprehensive Surveillance.

Ticks are obligate hematophagous arthropods of significant importance to human and veterinary medicine. They transmit a vast array of pathogens, including bacteria, viruses, protozoa, and helminths. Most epidemiological data on ticks and tick-borne pathogens (TBPs) in the West Indies are limited to common livestock pathogens such as Ehrlichia ruminantium, Babesia spp. (i.e., B. bovis and B. bigemina), and Anaplasma marginale, and less information is available on companion animal pathogens. Of note, human tick-borne diseases (TBDs) remain almost completely uncharacterized in the West Indies. Information on TBP presence in wildlife is also missing. Herein, we provide a comprehensive review of the ticks and TBPs affecting human and animal health in the Caribbean, and introduce the challenges associated with understanding TBD epidemiology and implementing successful TBD management in this region. In particular, we stress the need for innovative and versatile surveillance tools using high-throughput pathogen detection (e.g., high-throughput real-time microfluidic PCR). The use of such tools in large epidemiological surveys will likely improve TBD prevention and control programs in the Caribbean.

Occurrence of Leishmania chagasi, Trypanosoma cruzi, Babesia canis vogeli, Anaplasma platys, and Ehrlichia canis in canine blood donors / Ocorrência de Leishmania chagasi, Trypanosoma cruzi, Babesia canis vogeli, Anaplasma platys e Ehrlichia canis em cães doadores de sangue

The transfusion of blood components is common in a veterinary clinic; however, the safety of this therapeutic measure cannot always be guaranteed. Studies show a high risk of haemoparasite transmission during blood transfusion in canines. These parasites include Leishmania chagasi, Anaplasma platys, and Ehrlichia canis, which are endemic to the city of Cuiabá. This study aimed to evaluate the occurrence of L. chagasi, Trypanosoma cruzi, Babesia (canis) vogeli, A. platys, and E. canis in canine blood donor candidates, and identify possible factors associated with the infection of these agents. Sixty-six canines were evaluated using serologic and molecular tests, for the presence of the Leishmania species. While one canine sample showed a positive result for L. chagasi with indirect fluorescent antibody test, with titer of 1:40, and seven canine samples were positive using DPP, all other samples were negative when using PCR and ELISA. All canines were negative for T. cruzi when using PCR. The B. (c.) vogeli infection was identified in one canine and A. platys was identified in six canines. E. canis was identified in 17 canines, with a prevalence of 25.7%. There were no significant factors associated with the infection of the pathogens investigated. Given the observation of infection, even in the absence of clinical symptoms, emphasis must be placed on the need for the use of more sensitive and specific diagnostic methods for the screening of donor canines.(AU)

Administração de hemocomponentes é uma prática de rotina na clínica veterinária, entretanto, a segurança nessa medida terapêutica nem sempre é garantida. Estudos mostram risco elevado na transmissão de hemoparasitos pela transfusão sanguínea em cães. Dentre estes, Leishmania chagasi, Anaplasma platys e Ehrlichia canis são endêmicas no município de Cuiabá. Este estudo teve como objetivo avaliar a ocorrência de L. chagasi, Trypanosoma cruzi, Babesia (canis) vogeli, A. platys e E. canis em cães candidatos a doadores de sangue e identificar os possíveis fatores associados à infecção pelos referidos agentes. Dos 66 cães avaliados por exames sorológicos e moleculares para Leishmania sp, um cão reagiu na IFI na titulação 1:40 para L. chagasi., sete no DPP, e todos negativos na PCR e ELISA. Todos os cães foram negativos na PCR para T. cruzi. Foi identificada infecção por Babesia (c.) vogeli em apenas um cão e seis para A. platys. E. canis foi diagnosticado em 17 cães, com ocorrência de 25,7%. Não foram observados fatores associados significativamente à infecção pelos patógenos pesquisados. Diante da observação da infecção mesmo na ausência de alterações clínicas, ressalta-se o uso de métodos diagnósticos mais sensíveis e específicos na triagem de cães doadores.(AU)

Occurrence of Leishmania chagasi, Trypanosoma cruzi, Babesia canis vogeli, Anaplasma platys, and Ehrlichia canis in canine blood donors / Ocorrência de Leishmania chagasi, Trypanosoma cruzi, Babesia canis vogeli, Anaplasma platys e Ehrlichia canis em cães doadores de sangue

The transfusion of blood components is common in a veterinary clinic; however, the safety of this therapeutic measure cannot always be guaranteed. Studies show a high risk of haemoparasite transmission during blood transfusion in canines. These parasites include Leishmania chagasi, Anaplasma platys, and Ehrlichia canis, which are endemic to the city of Cuiabá. This study aimed to evaluate the occurrence of L. chagasi, Trypanosoma cruzi, Babesia (canis) vogeli, A. platys, and E. canis in canine blood donor candidates, and identify possible factors associated with the infection of these agents. Sixty-six canines were evaluated using serologic and molecular tests, for the presence of the Leishmania species. While one canine sample showed a positive result for L. chagasi with indirect fluorescent antibody test, with titer of 1:40, and seven canine samples were positive using DPP, all other samples were negative when using PCR and ELISA. All canines were negative for T. cruzi when using PCR. The B. (c.) vogeli infection was identified in one canine and A. platys was identified in six canines. E. canis was identified in 17 canines, with a prevalence of 25.7%. There were no significant factors associated with the infection of the pathogens investigated. Given the observation of infection, even in the absence of clinical symptoms, emphasis must be placed on the need for the use of more sensitive and specific diagnostic methods for the screening of donor canines.

Administração de hemocomponentes é uma prática de rotina na clínica veterinária, entretanto, a segurança nessa medida terapêutica nem sempre é garantida. Estudos mostram risco elevado na transmissão de hemoparasitos pela transfusão sanguínea em cães. Dentre estes, Leishmania chagasi, Anaplasma platys e Ehrlichia canis são endêmicas no município de Cuiabá. Este estudo teve como objetivo avaliar a ocorrência de L. chagasi, Trypanosoma cruzi, Babesia (canis) vogeli, A. platys e E. canis em cães candidatos a doadores de sangue e identificar os possíveis fatores associados à infecção pelos referidos agentes. Dos 66 cães avaliados por exames sorológicos e moleculares para Leishmania sp, um cão reagiu na IFI na titulação 1:40 para L. chagasi., sete no DPP, e todos negativos na PCR e ELISA. Todos os cães foram negativos na PCR para T. cruzi. Foi identificada infecção por Babesia (c.) vogeli em apenas um cão e seis para A. platys. E. canis foi diagnosticado em 17 cães, com ocorrência de 25,7%. Não foram observados fatores associados significativamente à infecção pelos patógenos pesquisados. Diante da observação da infecção mesmo na ausência de alterações clínicas, ressalta-se o uso de métodos diagnósticos mais sensíveis e específicos na triagem de cães doadores.

Hematological parameters and seroprevalence of Ehrlichia canis and Babesia vogeli in dogs / Parâmetros hematológicos e soroprevalência de Ehrlichia canis e Babesia vogeli em cães

The aim of this study was to determine the prevalence of anti-Ehrlichia canis and anti-Babesia vogeli IgG antibodies in dogs and correlate this prevalence with risk factors to evaluate the relation of serological status to hematological findings. Blood samples of dogs attended from September 2011 to March 2012 at the veterinary hospital of the Federal University of Lavras, Brazil, were analyzed using an indirect fluorescence antibody test (IFAT). Of the total 160 dog serum samples, 23.7% (38 dogs; CI95 17.7% 30.7%) were seropositive for E. canis, 40.0% (64 dogs; CI95 40.0% 59.2%) for B. vogeli, and 5.6% (9 dogs) for both hemoparasites. None of the epidemiological variables showed a significant association (P>0.05) with seropositivity to E. canis and B. vogeli. Dogs seropositive for E. canis showed lower values for hematocrit (P < 0.05). However, for the erythrogram, the platelet count, the leukogram and clinical signs, no significant difference (P>0.05) was observed between dogs that were seropositive and seronegative for E. canis or for B. vogeli. Serological results suggest that infection with E. canis and B. vogeli is endemic in the canine population in question, with a prevalence of the subclinical phase (asymptomatic) in dogs that are seropositive for ehrlichiosis or babesiosis.(AU)

O objetivo deste estudo foi determinar a prevalência de anticorpos IgG anti-Ehrlichia canis e anti-Babesia vogeli em cães e correlacionar com a prevalência e fatores de risco para avaliar a relação do estado sorológico com os achados hematológicos. Amostras de sangue de cães coletadas de setembro de 2011 a março de 2012, no Hospital Veterinário da Universidade Federal de Lavras, Brasil, foram analisadas usando o teste de reação de imunofluorescência indireta (RIFI). Do total de amostras de soro de 160 cães, 23,7% (38 cães; IC95 17,7% - 30,7%) foram soropositivos para E. canis, 40,0% (64 cães; IC95 40,0% - 59,2%) para B. vogeli, e 5,6% (9 cães) para os dois hemoparasitas. Nenhuma das variáveis epidemiológicas mostrou associação significativa (P> 0,05) com a soropositividade para E. canis e B. vogeli. Cães soropositivos para E. canis mostraram valores médios mais baixos para hematócrito (P < 0,05). No entanto, para o eritrograma, a contagem de plaquetas, o leucograma e os sinais clínicos, nenhuma diferença significativa (P> 0,05) foi observada entre cães soropositivos e soronegativos para E. canis ou para B. vogeli. Os resultados sorológicos deste estudo sugerem que a infecção por E. canis e B. vogeli é endêmica na população canina em questão, com uma prevalência da fase subclínica (assintomática) em cães soropositivos para erliquiose ou babesiose.(AU)

Hematological parameters and seroprevalence of Ehrlichia canis and Babesia vogeli in dogs / Parâmetros hematológicos e soroprevalência de Ehrlichia canis e Babesia vogeli em cães

The aim of this study was to determine the prevalence of anti-Ehrlichia canis and anti-Babesia vogeli IgG antibodies in dogs and correlate this prevalence with risk factors to evaluate the relation of serological status to hematological findings. Blood samples of dogs attended from September 2011 to March 2012 at the veterinary hospital of the Federal University of Lavras, Brazil, were analyzed using an indirect fluorescence antibody test (IFAT). Of the total 160 dog serum samples, 23.7% (38 dogs; CI95 17.7% 30.7%) were seropositive for E. canis, 40.0% (64 dogs; CI95 40.0% 59.2%) for B. vogeli, and 5.6% (9 dogs) for both hemoparasites. None of the epidemiological variables showed a significant association (P>0.05) with seropositivity to E. canis and B. vogeli. Dogs seropositive for E. canis showed lower values for hematocrit (P 0.05) was observed between dogs that were seropositive and seronegative for E. canis or for B. vogeli. Serological results suggest that infection with E. canis and B. vogeli is endemic in the canine population in question, with a prevalence of the subclinical phase (asymptomatic) in dogs that are seropositive for ehrlichiosis or babesiosis.

O objetivo deste estudo foi determinar a prevalência de anticorpos IgG anti-Ehrlichia canis e anti-Babesia vogeli em cães e correlacionar com a prevalência e fatores de risco para avaliar a relação do estado sorológico com os achados hematológicos. Amostras de sangue de cães coletadas de setembro de 2011 a março de 2012, no Hospital Veterinário da Universidade Federal de Lavras, Brasil, foram analisadas usando o teste de reação de imunofluorescência indireta (RIFI). Do total de amostras de soro de 160 cães, 23,7% (38 cães; IC95 17,7% - 30,7%) foram soropositivos para E. canis, 40,0% (64 cães; IC95 40,0% - 59,2%) para B. vogeli, e 5,6% (9 cães) para os dois hemoparasitas. Nenhuma das variáveis epidemiológicas mostrou associação significativa (P> 0,05) com a soropositividade para E. canis e B. vogeli. Cães soropositivos para E. canis mostraram valores médios mais baixos para hematócrito (P 0,05) foi observada entre cães soropositivos e soronegativos para E. canis ou para B. vogeli. Os resultados sorológicos deste estudo sugerem que a infecção por E. canis e B. vogeli é endêmica na população canina em questão, com uma prevalência da fase subclínica (assintomática) em cães soropositivos para erliquiose ou babesiose.

Clinical and hematological evaluation of Rangelia vitalii-naturally infected dogs in southeastern Brazil

Rangelia vitalii, a tick-borne piroplasm that infects dogs, has been recently molecularly characterized in Brazil, Uruguay and Argentina. Studies on molecular characterization of these piroplasms in different Brazilian regions are scarce. The aim of this study was to evaluate clinical and hematological changes in dogs caused by R. vitalii in the mountainous region of the state of Rio de Janeiro. Blood samples from 36 dogs were evaluated for piroplasms and hematological disorders using light microscopy and molecular analysis. Blood samples from all the animals included in this study were confirmed to be positive for R. vitalii through genetic sequencing. Clinical signspresented by 24 of the 36 dogs of the study were evaluated during appointments or hospitalization within private practice. The most frequent clinical disorders in these dogs that were naturally infected with R. vitalii were fever, spontaneous cutaneous bleeding and diarrhea. Normochromic non-regenerative anemia and thrombocytopenia were the most common hematological disorders in these R. vitalii-positive dogs and therefore should be considered in hematological evaluations on suspected cases.(AU)

Rangelia vitalii, um piroplasma transmitido por carrapatos que infecta cães, foi sendo recentemente caracterizado molecularmente no Brasil, Uruguai e Argentina. Nas diferentes regiões brasileiras são escassos os estudos acerca da caracterização molecular destes piroplasmídeos. O objetivo deste estudo foi avaliar as alterações clínicas e hematológicas em cães causadas por R. vitalii na região serrana do Rio de Janeiro. Amostras de sangue total de 36 cães foram examinadas quanto à presença de piroplasmas pela microscopia de luz, alterações hematológicas e análise molecular. Todos os cães do presente estudo foram positivos para R. vitalii através do sequenciamento genético. Dos 36 animais positivos para R. vitalii, 24 foram avaliados clinicamente. As alterações mais frequentemente observadas foram febre, sangramento cutâneo espontâneo e diarréia. Anemia normocítica normocrômica arregenerativa e trombocitopenia foram as alterações hematológicas mais observadas em cães positivos para R. vitalii, devendo ser consideradas na avaliação hematológica de cães suspeitos.(AU)

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

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.(AU)

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.(AU)

Diferenciação hematológica e bioquímica de cães infectados por Leishmania infantum, Babesia spp., Dirofilaria spp. e vírus da Cinomose / Hematologic and biochemical differentiation of dogs infected with Leishmania infantum, Babesia spp., Dirofilaria spp. and Cinomosis virus

Patógenos virais, protozoários e helmintoícos podem ser transmitidos simultaneamente ao mesmo hospedeiro canino, através de vetores biológicos. A avaliação de parâmetros hematobioquímicos pode fornecer dados acerca da fisiopatologia das coinfecções. Nesse contexto, objetivou-se avaliar o papel da coinfecção natural sobre o perfil hematológico e bioquímico de cães, naturalmente infectados por Leishmania infantum. Para tanto, foram utilizados 31 cães, distribuídos entre grupo controle (Grupo A, n=10) e grupo infectado (Grupo B, n=21), que foi subdividido em cães mono-infectados com L. infantum (Grupo B1, n=14) e co-infectados com L. infantum e Babesia spp., Dirofilaria spp. ou vírus da cinomose canina (Grupo B2, n=7). Amostras de sangue e soro foram analisadas, respectivamente, para avaliação de parâmetros hematológicos e dosagens bioquímicas de AST, ALT, ureia, creatinina e proteína total. Previamente, os animais foram avaliados por métodos sorológicos e parasitológicos, para confirmar as coinfecções. O protocolo experimental foi aprovado pelo CEUA/UECE (4609061/2014). Os resultados foram expressos em média±desvio padrão e foram analisados pelo teste t-Student não-pareado com p≤0,05. Hemácias, hemoglobina, hematócrito, plaquetas, eosinófilos e linfócitos apresentaram valores reduzidos (p≤0,05) nos Grupos B e B2, quando comparados com o Grupo A; enquanto que os teores de AST foram maiores (p≤0,05), na mesma comparação. O valor de ureia foi maior (p≤0,05) no Grupo B e menor no Grupo B2, quando comparado com o Grupo A. Os valores de ALT foram maiores (p≤0,05) no Grupo B1, em relação ao B2. As co-infecções influenciam nos parâmetros hematológicos e os níveis séricos de ALT, AST e ureia são importantes na avaliação bioquímica de cães acometidos por Leishmania infantum, em relação aos animais infectados...

Virus, protozoa and helminths may be transmitted simultaneously to the same canine host through biological vectors. The evaluation of blood-biochemical parameters can provide data about the pathophysiology of coinfection. In this context, the objective was to evaluate the role of natural coinfection on hematological and biochemical profile of dogs naturally infected by Leishmania infantum. To this end, 31 dogs were randomly divided between control group (Group A, n=10) and infected group (group B, n=21), which was divided into dogs mono-infected with L. infantum (Group B1, n=14) and dogs co-infected with L. infantum and Dirofilaria spp, Babesia spp or canine distemper virus (B2 Group, n=7). Blood and serum samples were analyzed, respectively, for hematological and biochemical levels of AST, ALT, urea, creatinine, and total protein. Previously, animals were evaluated by serological and parasitological methods to confirm the co-infections. The experimental protocol was approved by CEUA / UECE (4609061/2014). Results were expressed as mean ± standard deviation and were analyzed by unpaired t-test with p≤0.05. Erythrocytes, hemoglobin, hematocrit and platelets showed reduced values (p≤0.05) in group B and B2 compared with group A, while AST levels were higher (p≤0.05) in the same comparison. The amount of urea was higher (p≤0.05) in Group B and lower in Group B2 when compared to Group A. ALT levels were higher (p≤0.05) in Group B1 relative to B2. It is concluded that coinfection influence on hematological parameters and serum levels of ALT, AST and urea are important in biochemical evaluation of affected dogs by L. infantum.

Diferenciação hematológica e bioquímica de cães infectados por Leishmania infantum, Babesia spp., Dirofilaria spp. e vírus da Cinomose / Hematologic and biochemical differentiation of dogs infected with Leishmania infantum, Babesia spp., Dirofilaria spp. and Cinomosis virus

Patógenos virais, protozoários e helmintoícos podem ser transmitidos simultaneamente ao mesmo hospedeiro canino, através de vetores biológicos. A avaliação de parâmetros hematobioquímicos pode fornecer dados acerca da fisiopatologia das coinfecções. Nesse contexto, objetivou-se avaliar o papel da coinfecção natural sobre o perfil hematológico e bioquímico de cães, naturalmente infectados por Leishmania infantum. Para tanto, foram utilizados 31 cães, distribuídos entre grupo controle (Grupo A, n=10) e grupo infectado (Grupo B, n=21), que foi subdividido em cães mono-infectados com L. infantum (Grupo B1, n=14) e co-infectados com L. infantum e Babesia spp., Dirofilaria spp. ou vírus da cinomose canina (Grupo B2, n=7). Amostras de sangue e soro foram analisadas, respectivamente, para avaliação de parâmetros hematológicos e dosagens bioquímicas de AST, ALT, ureia, creatinina e proteína total. Previamente, os animais foram avaliados por métodos sorológicos e parasitológicos, para confirmar as coinfecções. O protocolo experimental foi aprovado pelo CEUA/UECE (4609061/2014). Os resultados foram expressos em média±desvio padrão e foram analisados pelo teste t-Student não-pareado com p≤0,05. Hemácias, hemoglobina, hematócrito, plaquetas, eosinófilos e linfócitos apresentaram valores reduzidos (p≤0,05) nos Grupos B e B2, quando comparados com o Grupo A; enquanto que os teores de AST foram maiores (p≤0,05), na mesma comparação. O valor de ureia foi maior (p≤0,05) no Grupo B e menor no Grupo B2, quando comparado com o Grupo A. Os valores de ALT foram maiores (p≤0,05) no Grupo B1, em relação ao B2. As co-infecções influenciam nos parâmetros hematológicos e os níveis séricos de ALT, AST e ureia são importantes na avaliação bioquímica de cães acometidos por Leishmania infantum, em relação aos animais infectados...(AU)

Virus, protozoa and helminths may be transmitted simultaneously to the same canine host through biological vectors. The evaluation of blood-biochemical parameters can provide data about the pathophysiology of coinfection. In this context, the objective was to evaluate the role of natural coinfection on hematological and biochemical profile of dogs naturally infected by Leishmania infantum. To this end, 31 dogs were randomly divided between control group (Group A, n=10) and infected group (group B, n=21), which was divided into dogs mono-infected with L. infantum (Group B1, n=14) and dogs co-infected with L. infantum and Dirofilaria spp, Babesia spp or canine distemper virus (B2 Group, n=7). Blood and serum samples were analyzed, respectively, for hematological and biochemical levels of AST, ALT, urea, creatinine, and total protein. Previously, animals were evaluated by serological and parasitological methods to confirm the co-infections. The experimental protocol was approved by CEUA / UECE (4609061/2014). Results were expressed as mean ± standard deviation and were analyzed by unpaired t-test with p≤0.05. Erythrocytes, hemoglobin, hematocrit and platelets showed reduced values (p≤0.05) in group B and B2 compared with group A, while AST levels were higher (p≤0.05) in the same comparison. The amount of urea was higher (p≤0.05) in Group B and lower in Group B2 when compared to Group A. ALT levels were higher (p≤0.05) in Group B1 relative to B2. It is concluded that coinfection influence on hematological parameters and serum levels of ALT, AST and urea are important in biochemical evaluation of affected dogs by L. infantum.(AU)

Oxidative stress control by apicomplexan parasites.

Apicomplexan parasites cause infectious diseases that are either a severe public health problem or an economic burden. In this paper we will shed light on how oxidative stress can influence the host-pathogen relationship by focusing on three major diseases: babesiosis, coccidiosis, and toxoplasmosis.

Typification of virulent and low virulence Babesia bigemina clones by 18S rRNA and rap-1c.

The population structure of original Babesia bigemina isolates and reference strains with a defined phenotypic profile was assessed using 18S rRNA and rap-1c genes. Two reference strains, BbiS2P-c (virulent) and BbiS1A-c (low virulence), were biologically cloned in vitro. The virulence profile of the strains and clones was assessed in vivo. One fully virulent and one low-virulence clone were mixed in identical proportions to evaluate their growth efficiency in vitro. Each clone was differentiated by two microsatellites and the gene gp45. The 18S rRNA and rap-1c genes sequences from B. bigemina biological clones and their parental strains, multiplied exclusively in vivo or in vitro, were compared with strain JG-29. The virulence of clones derived from the BbiS2P-c strain was variable. Virulent clone Bbi9P1 grew more efficiently in vitro than did the low-virulence clone Bbi2A1. The haplotypes generated by the nucleotide polymorphism, localized in the V4 region of the 18S rRNA, allowed the identification of three genotypes. The rap-1c haplotypes allowed defining four genotypes. Parental and original strains were defined by multiple haplotypes identified in both genes. The rap-1c gene, analyzed by high-resolution melting (HRM), allowed discrimination between two genotypes according to their phenotype, and both were different from JG-29. B. bigemina biological clones made it possible to define the population structure of isolates and strains. The polymorphic regions of the 18S rRNA and rap-1c genes allowed the identification of different subpopulations within original B. bigemina isolates by the definition of several haplotypes and the differentiation of fully virulent from low virulence clones.

Babesia: a world emerging.

Babesia are tick-transmitted hemoprotozooans that infect mammals and birds, and which are acknowledged for their major impact on farm and pet animal health and associated economic costs worldwide. Additionally, Babesia infections of wildlife can be fatal if associated with stressful management practices; and human babesiosis, also transmitted by blood transfusion, is an increasing public-health concern. Due to the huge diversity of species reported to serve as Babesia hosts, all vertebrates might be potential carriers, as long as they are adequate hosts for Babesia-vector ticks. We here provide a comprehensive overview of the most relevant Babesia species, and a discussion of the classical taxonomic criteria. Babesia, Cytauxzoon and Theileria parasites are closely related and collectively referred to as piroplasmids. A possible scenario for the history of piroplasmids is presented in the context of recent findings, and its implications for future research avenues are outlined. Phylogenetic trees of all available 18S rRNA and hsp70 genes were generated, based on which we present a thoroughly revised molecular classification, comprising five monophyletic Babesia lineages, one Cytauxzoon clade, and one Theileria clade. Updated 18S rRNA and beta-tubulin gene trees of the B. microti isolates agree with those previously reported. To reconcile estimates of the origin of piroplasmids and ticks (~300 Ma, respectively), and mammalian radiation (60 Ma), we hypothesize that the dixenous piroplasmid life cycle evolved with the origin of ticks. Thus, the observed time gap between tick origin and mammalian radiation indicates the existence of hitherto unknown piroplasmid lineages and/or species in extant vertebrate taxa, including reptiles and possibly amphibians. The development and current status of the molecular taxonomy of Babesia, with emphasis on human-infecting species, is discussed. Finally, recent results from population genetic studies of Babesia parasites, and their implications for the development of pathogenicity, drug resistance and vaccines, are summarized.

Characterization and transcriptional analysis of the promoter region of the Duffy blood group, chemokine receptor (DARC) gene in cattle.

The Duffy antigen is the only receptor for Plasmodium vivax, a hemoparasite of the phylum Apicomplexa and the cause of vivax malaria in humans. Resistance to this parasite in the majority of black African individuals and their descendents is due to a mutation in the gene promoter region, which blocks its transcription on erythrocytes. Regarding bovine babesiosis, it is known that taurine breeds are more susceptible to parasite infection than zebuine breeds. In order to verify whether the same human resistance occurs in bovine, the 5' flanking region of the DARC gene was isolated and characterized in Bos indicus and Bos taurus. Four single nucleotide polymorphisms were identified and genotyped (SNP1: EF_647729.1:g.91C>T; SNP2: EF_647729.1:g.405C>T; SNP3: EF_647729.1: g.433A>G and SNP4: EF_647729.1:g.588A>G), which showed significant frequency differences among 99 bovines of each species (n=198). Characterization of the isolated region revealed the presence of 6 putative haplotypes, 14 genotypes, which are formed by haplotypes, and numerous putative transcription factor binding sites. Only the thymine presence on SNPs 1 and 2, more common in B. indicus, was observed to alter some of the sites in this region. Despite this fact, analyses through real-time PCR on bovines that present the most common homozygote genotypes of each species, which contrast for all the polymorphism, revealed no difference on the DARC gene transcription. Thus, in principle, it was concluded that the polymorphisms identified would not be useful as molecular markers in an improvement program for resistance to babesiosis.