Dogs naturally infected by Rangelia vitalii, Babesia canis vogeli, and Ehrlichia canis in São Paulo, Brazil / Cães naturalmente infectados por Rangelia vitalii, Babesia canis vogeli, e Ehrlichia canis em São Paulo, Brasil

Several agents can cause hemoparasitic diseases in dogs, and blood-sucking arthropods transmit these diseases. These agents can cause several clinical manifestations and, in some cases, can kill the host. Because these agents are essential in animal health, this study aims to detect the frequency of Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, and Rangelia vitalii by real-time PCR and Babesia vogeli in dogs in the southern region of the city of São Paulo, São Paulo. Of the 98 dog samples, 18 (18.4%) tested positive with real-time polymerase chain reaction for at least one studied agent. Of these 18 samples, 17 tested positive for a single agent (11.2% for B. canis vogeli, 1.02% for R. vitalii, and 5.1% for E. canis), and one showed co-infection with B. canis vogeli and R. vitalii. The results demonstrate the presence of hemoparasites in the studied animals, which can influence the quality and life expectancy of these animals. The Rangeliadetection warns small animal clinicians to include it as a differential diagnosis for hemoparasitosis.(AU)

As hemoparasitoses em cães podem ser causadas por diversos agentes, sendo essas doenças transmitidas por artrópodes hematófagos. Esses agentes podem causar diversas manifestações clínicas e, em alguns casos, podem matar o hospedeiro. Este estudo teve como objetivo detectar por PCR em tempo real a frequência de Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, Rangelia vitalii e Babesia canis vogeli em amostras de cães da zona sul da cidade de São Paulo, Brasil. Das 98 amostras de cães, 18 (18,4%) testaram positivo com reação em cadeia da polimerase em tempo real para pelo menos um agente estudado. Destas 18 amostras, 17 testaram positivo para um único agente (11,2% para B. canis vogeli, 1,02% para R. vitalii e 5,1% para E. canis), e uma apresentou coinfecção com B. canis vogeli e R. vitalii. Os resultados demonstram a presença de hemoparasitas nos animais estudados, o que pode influenciar a qualidade e a expectativa de vida desses animais. Além disso, é o primeiro relato da detecção de R. vitalli na zona sul de São Paulo e serve de alerta para os clínicos de pequenos animais incluírem esse agente como diagnóstico diferencial para as hemoparasitoses.(AU)

Hemoparasites in Didelphis aurita from Seropédica, Rio de Janeiro state, Brazil / Hemoparasitos em Didelphis aurita capturados em Seropédica, estado do Rio de Janeiro, Brasi

The present study aimed to use polymerase chain reaction (PCR) to detect species of the order Piroplasmida, such as Anaplasma spp., Borrelia spp., and Ehrlichia spp., circulating in the blood of Didelphis aurita in a peridomiciliary environment. Blood samples collected from big-eared opossum (Didelphis aurita) were screened for hemoparasites using PCR. The extracted DNA was tested for tick-borne hemoparasites. We were unable to detect hemoparasites, such as Ehrlichia spp., Babesia spp., Anaplasma spp., and Borrelia spp. Theileria DNA was detected in only one sample screened using PCR for an approximately 650-base pair fragment of the 18S rRNA gene. Sequencing and BLAST analysis of a subset of the PCR amplicons revealed 97% (535/553 bp) identity with Theileria bicornis. The detection of Theileria sp. in D. aurita challenges us to pursue more in-depth studies of marsupial piroplasmosids and to evaluate the morphological aspects of the findings and their possible involvement in zoonoses.

O objetivo do presente estudo foi utilizar a reação em cadeia da polimerase (PCR) para detectar espécies da ordem Piroplasmida, como Anaplasma spp., Borrelia spp. e Ehrlichia spp., circulando no sangue de Didelphis aurita em ambiente peridomiciliar. Amostras de sangue coletadas de gambá da orelha preta (Didelphis aurita) foram triadas para hemoparasitas por meio da PCR. O DNA extraído foi testado para alguns hemoparasitas transmitidos por carrapatos. Não conseguimos detectar hemoparasitos, como Ehrlichia spp., Babesia spp., Anaplasma spp. e Borrelia spp. O DNA de Theileria foi detectado apenas em uma amostra. Um fragmento de ~650 pares de bases do gene 18S rRNA foi sequenciado. e a análise pelo BLAST (um subconjunto de amplicons de PCR) revelou uma identidade de 97% (535/553 pb) com Theileria bicornis. A detecção de Theileria sp. in D. aurita nos desafia a aprofundar estudos sobre piroplasmosídeos marsupiais e avaliar os aspectos morfológicos dos achados e seu possível envolvimento com zoonoses.

Molecular diagnosis and risk factors of canine ehrlichiosis in the municipality of Itabuna-Bahia, Brazil / Diagnóstico molecular e fatores de risco da erliquiose canina no município de Itabuna-Bahia

Ehrlichiosis is an emerging zoonosis worldwide and has had several adverse effects on public health. Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, has the tick Rhipicephalus sanguineus as the vector. The main clinical signs in affected dogs are fever, apathy, anorexia, weight loss, and neurological signs. The diagnosis is made through the association of clinical signs with parasitological, serological, and molecular tests. The aim of this study was to evaluate the occurrence of E. canis infection in dogs from the city of Itabuna-Bahia, as well as to identify the risk factors related to infection. For this, 405 dogs from the Center for Zoonoses Control (CCZ), non-governmental organizations (NGOs), and dogs domiciled and semi-domiciled in the city of Itabuna, southern Bahia, were evaluated. After initial physical evaluation of the dogs, blood samples were collected by venipuncture for subsequent DNA extraction and E. canis testing using the nested Polymerase Chain Reaction (nested-PCR) technique. In addition, an epidemiological questionnaire that included questions related to the animals was administered to the dog owners to identify the risk factors for exposure to the etiological agent and to the vector. Approximately 17% of the dogs in the municipality of Itabuna-Bahia tested positive for E. canis by nested-PCR, a result higher than that found in other studies conducted in the same municipality. Among the factors associated with E. canis infection, contact with other dogs (p = 0.0226) was an important factor for the dissemination of CME, since dogs are reported to be reservoirs of E. canis. Male dogs (p = 0.0016) presented lower risk for E. canis infection. Other studies, however, describe no association between animal gender and infection by E. canis. Preventive measures to reduce exposure to the vector of ehrlichiosis are necessary.

A erliquiose é uma zoonose emergente em todo o mundo e tem acarretado diversos transtornos para a saúde pública. A erliquiose monocítica canina (EMC), causada pela Ehrlichia canis, tem como vetor o carrapato Rhipicephalus sanguineus. Os principais sinais clínicos nos cães afetados são febre, apatia, anorexia, perda de peso e sinais neurológicos. O diagnóstico é feito através da associação dos sinais clínicos, exames parasitológicos, sorológicos e moleculares. Objetivou-se através deste estudo avaliar a ocorrência de infecção por E. canis em cães do município de Itabuna-Bahia, bem como identificar os fatores de risco relacionados à infecção. Para tanto, foram avaliados 405 cães provenientes do Centro de Controle de Zoonoses (CCZ), Organizações não governamentais (ONGs), cães domiciliados e semi-domiciliados da cidade de Itabuna-Bahia. Após avaliação física inicial dos cães, procedeu-se em seguida a coleta de amostras de sangue por punção venosa, para posterior extração de DNA e pesquisa de E. canis pela técnica de nested-Reação em Cadeia de Polimerase (nested-PCR). Adicionalmente, um questionário epidemiológico foi aplicado junto aos responsáveis, no qual constavam questões relacionadas aos animais, com a finalidade de identificar os fatores de risco de exposição destes ao agente etiológico e ao vetor. Em suma, este estudo mostrou que aproximadamente 17% dos cães do município de Itabuna-Bahia foram positivos para E. canis pela nested-PCR, resultado superior ao encontrado em outros estudos realizados no mesmo município. Dos fatores associados à infecção por E. canis, foi significativo o contato com outros cães (p=0,0226), um fator importante para a disseminação da EMC, pois os cães são relatados como reservatórios da E. canis. O gênero macho (p=0,0016) apresentou menor risco para a infecção por E. canis. Outros estudos, no entanto, descrevem que não há nenhuma associação entre o gênero do animal e a infecção por...

Molecular diagnosis and risk factors of canine ehrlichiosis in the municipality of Itabuna-Bahia, Brazil / Diagnóstico molecular e fatores de risco da erliquiose canina no município de Itabuna-Bahia

Ehrlichiosis is an emerging zoonosis worldwide and has had several adverse effects on public health. Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, has the tick Rhipicephalus sanguineus as the vector. The main clinical signs in affected dogs are fever, apathy, anorexia, weight loss, and neurological signs. The diagnosis is made through the association of clinical signs with parasitological, serological, and molecular tests. The aim of this study was to evaluate the occurrence of E. canis infection in dogs from the city of Itabuna-Bahia, as well as to identify the risk factors related to infection. For this, 405 dogs from the Center for Zoonoses Control (CCZ), non-governmental organizations (NGOs), and dogs domiciled and semi-domiciled in the city of Itabuna, southern Bahia, were evaluated. After initial physical evaluation of the dogs, blood samples were collected by venipuncture for subsequent DNA extraction and E. canis testing using the nested Polymerase Chain Reaction (nested-PCR) technique. In addition, an epidemiological questionnaire that included questions related to the animals was administered to the dog owners to identify the risk factors for exposure to the etiological agent and to the vector. Approximately 17% of the dogs in the municipality of Itabuna-Bahia tested positive for E. canis by nested-PCR, a result higher than that found in other studies conducted in the same municipality. Among the factors associated with E. canis infection, contact with other dogs (p = 0.0226) was an important factor for the dissemination of CME, since dogs are reported to be reservoirs of E. canis. Male dogs (p = 0.0016) presented lower risk for E. canis infection. Other studies, however, describe no association between animal gender and infection by E. canis. Preventive measures to reduce exposure to the vector of ehrlichiosis are necessary.(AU)

A erliquiose é uma zoonose emergente em todo o mundo e tem acarretado diversos transtornos para a saúde pública. A erliquiose monocítica canina (EMC), causada pela Ehrlichia canis, tem como vetor o carrapato Rhipicephalus sanguineus. Os principais sinais clínicos nos cães afetados são febre, apatia, anorexia, perda de peso e sinais neurológicos. O diagnóstico é feito através da associação dos sinais clínicos, exames parasitológicos, sorológicos e moleculares. Objetivou-se através deste estudo avaliar a ocorrência de infecção por E. canis em cães do município de Itabuna-Bahia, bem como identificar os fatores de risco relacionados à infecção. Para tanto, foram avaliados 405 cães provenientes do Centro de Controle de Zoonoses (CCZ), Organizações não governamentais (ONGs), cães domiciliados e semi-domiciliados da cidade de Itabuna-Bahia. Após avaliação física inicial dos cães, procedeu-se em seguida a coleta de amostras de sangue por punção venosa, para posterior extração de DNA e pesquisa de E. canis pela técnica de nested-Reação em Cadeia de Polimerase (nested-PCR). Adicionalmente, um questionário epidemiológico foi aplicado junto aos responsáveis, no qual constavam questões relacionadas aos animais, com a finalidade de identificar os fatores de risco de exposição destes ao agente etiológico e ao vetor. Em suma, este estudo mostrou que aproximadamente 17% dos cães do município de Itabuna-Bahia foram positivos para E. canis pela nested-PCR, resultado superior ao encontrado em outros estudos realizados no mesmo município. Dos fatores associados à infecção por E. canis, foi significativo o contato com outros cães (p=0,0226), um fator importante para a disseminação da EMC, pois os cães são relatados como reservatórios da E. canis. O gênero macho (p=0,0016) apresentou menor risco para a infecção por E. canis. Outros estudos, no entanto, descrevem que não há nenhuma associação entre o gênero do animal e a infecção por...(AU)

Interpretation of coagulation tendency contributing to thrombosis in vector-borne diseases (ehrlichiosis, anaplasmosis, leishmaniosis, and dirofilariasis) among dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs. Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid quantitave test analyser. Subgroups were formed according to the number of cases and the distribution of vector-borne agent. Statistically significant decreased PLT count was found in dogs mono infected with ehrlichiosis compared to healthy dogs (P < 0.001). Changes in mean PT value in the studied animals did not show statistically significant differences among the groups (P > 0.05). APTT values in the ehrlichiosis mono infection group were significantly higher than that of the healthy control (P < 0.01). A significant increase in FIB levels were detected […](AU)

Interpretation of coagulation tendency contributing to thrombosis in vector-borne diseases (ehrlichiosis, anaplasmosis, leishmaniosis, and dirofilariasis) among dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present author’s knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs. Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid quantitave test analyser. Subgroups were formed according to the number of cases and the distribution of vector-borne agent. Statistically significant decreased PLT count was found in dogs mono infected with ehrlichiosis compared to healthy dogs (P 0.05). APTT values in the ehrlichiosis mono infection group were significantly higher than that of the healthy control (P < 0.01). A significant increase in FIB levels were detected […]

Interpretation of Coagulation Tendency Contributing to Thrombosis in Vector-Borne Diseases (Ehrlichiosis, Anaplasmosis, Leishmaniosis, and Dirofilariasis) among Dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs.Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid qu

Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil / Detecção molecular e serológica de Theileria equi, Babesia caballi e Anaplasma phagocytophilum em equinos e carrapatos no Maranhão

Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT) and blood samples and ticks to Polymerase Chain Reaction (PCR) to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. It was detected 2.35% (4/170) and 0.59% (1/170) positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05) was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.(AU)

A piroplasmose equina é uma doença transmitida por carrapatos causada pelos protozoários intraeritrocitários Babesia caballi e Theileria equi. É relatada como uma doença parasitária comum em equinos. Além disso, Anaplasma phagocytophilum, o agente causal da ehrlichiose granulocítica, causa uma doença sazonal em equinos. Ambas as doenças, podem ser prejudiciais para a saúde animal. Nesse sentido, amostras de sangue e carrapatos foram coletadas de 97 cavalos criados na microrregião da Baixada Maranhense, estado do Maranhão, Brasil. As amostras de soro foram submetidas ao Teste de Imunofluorescência Indireta (RIFI) e amostras de sangue e os carrapatos a Reação da Polimerase em Cadeia (PCR) para avaliar a infecção por Theileria equi, Babesia caballi e Anaplasma phagocytophilum. A prevalência foi de 38,14%, 18,55% e 11,34% para T. equi, B. caballi e A. phagocytophilum, respectivamente. Os resultados da PCR para as amostras de sangue demonstraram 13,40% e 3,09% de positividade para T. equi e B. caballi, respectivamente. Um total de 170 specimens de carrapatos foi coletado e foram identificados Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. Obteve-se 2,35% (4/170) e 0,59% (1/170) positivos por PCR para T. equi e B. caballi, respectivamente. Todas as amostras foram negativas para A. phagocytophilum. Não houve diferença estatística significativa (p>0.05) em relação ao sexo, idade, uso de ectoparasiticida e presença de carrapatos. A análise BLASTn das amostras sequenciadas para gene 18S rRNA indicaram 97 a 100% de similaridade com T. equi e 98-100% com B. caballi no GenBank. Análises genéticas classificaram as sequencias obtidas no mesmo clado que T. equi e B. caballi, respectivamente. Podemos concluir que estes patógenos estão circulando na área de estudo.(AU)

Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil / Detecção molecular e serológica de Theileria equi, Babesia caballi e Anaplasma phagocytophilum em equinos e carrapatos no Maranhão

Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT) and blood samples and ticks to Polymerase Chain Reaction (PCR) to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. It was detected 2.35% (4/170) and 0.59% (1/170) positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05) was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.(AU)

A piroplasmose equina é uma doença transmitida por carrapatos causada pelos protozoários intraeritrocitários Babesia caballi e Theileria equi. É relatada como uma doença parasitária comum em equinos. Além disso, Anaplasma phagocytophilum, o agente causal da ehrlichiose granulocítica, causa uma doença sazonal em equinos. Ambas as doenças, podem ser prejudiciais para a saúde animal. Nesse sentido, amostras de sangue e carrapatos foram coletadas de 97 cavalos criados na microrregião da Baixada Maranhense, estado do Maranhão, Brasil. As amostras de soro foram submetidas ao Teste de Imunofluorescência Indireta (RIFI) e amostras de sangue e os carrapatos a Reação da Polimerase em Cadeia (PCR) para avaliar a infecção por Theileria equi, Babesia caballi e Anaplasma phagocytophilum. A prevalência foi de 38,14%, 18,55% e 11,34% para T. equi, B. caballi e A. phagocytophilum, respectivamente. Os resultados da PCR para as amostras de sangue demonstraram 13,40% e 3,09% de positividade para T. equi e B. caballi, respectivamente. Um total de 170 specimens de carrapatos foi coletado e foram identificados Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus) microplus. Obteve-se 2,35% (4/170) e 0,59% (1/170) positivos por PCR para T. equi e B. caballi, respectivamente. Todas as amostras foram negativas para A. phagocytophilum. Não houve diferença estatística significativa (p>0.05) em relação ao sexo, idade, uso de ectoparasiticida e presença de carrapatos. A análise BLASTn das amostras sequenciadas para gene 18S rRNA indicaram 97 a 100% de similaridade com T. equi e 98-100% com B. caballi no GenBank. Análises genéticas classificaram as sequencias obtidas no mesmo clado que T. equi e B. caballi, respectivamente. Podemos concluir que estes patógenos estão circulando na área de estudo.(AU)

Rickettsia parkeri em focos de febre maculosa no bioma Pampa brasileiro

A febre maculosa é uma doença febril aguda, com sintomatologia inespecífica, causada por bactérias do gênero Rickettsia e transmitida por carrapatos. Nesse trabalho relatamos a investigação eco-epidemiológica de dois casos da doença ocorridos no bioma Pampa brasileiro, nas cidades de Rosário do Sul em 2011, e Toropi, área de transição com Mata Atlântica, em 2014. Foram coletados carrapatos das duas áreas de foco, de capivaras, cães e do ambiente. Em Rosário do Sul foram coletadas também amostras de sangue dos cães da propriedade foco e vizinhança. Realizaram-se ainda, nas propriedades em que foram coletadas amostras, entrevistas que abrangiam perguntas sobre carrapatos e febre maculosa. Todos os carrapatos coletados foram processados individualmente para obtenção do DNA genômico. A pesquisa de Rickettsia spp. foi realizada por reação em cadeia da polimerase (PCR), utilizando primers específicos para fragmentos rickettsiais dos genes gltA, ompA e htrA. Nas amostras de soro canino foram pesquisados anticorpos contra Rickettsia spp. através da reação de imunofluorescência indireta (RIFI). Na investigação realizada no município de Rosário do Sul foram encontradas as seguintes espécies de carrapatos: 251 Amblyomma dubitatum em carcaças de capivaras e no ambiente; 60 larvas de Amblyomma spp. no ambiente; 62 adultos de Amblyomma tigrinum, dois Amblyomma aureolatum e quatro Rhipicephalus sanguineus nos cães da propriedade foco e vizinhança. Amostras de 24 A. tigrinum (38%) e uma de Rh. sanguineus foram positivas em todas as análises de PCR para Rickettsia spp. (gltA, ompA e htrA). No município de Toropi foram coletados 18 A. dubitatum de carcaças de capivaras, dos quais quatro foram positivos para presença de DNA de Rickettsia spp. Também foram testados 210 A. dubitatum obtidos de capivaras mortas em rodovias de outras localidades do bioma Pampa, nenhum deles foi positivo. Todas as amostras positivas (Rosário do Sul e Toropi) mostraram 100% de identidade com sequências de Rickettsia parkeri cepa Portsmouth. Na sorologia dos cães, todas as amostras (100%) continham anticorpos que reagiram pelo menos para uma espécie de Rickettsia, com títulos variando de 128 a 16384. Foi possível determinar R. parkeri como provável agente responsável pela infecção natural em 12 amostras (66,66%). Nas entrevistas observamos que o hábito de caça é frequente e os cães das propriedades são criados livres em contato com a mata, sendo observada alta taxa de parasitismo por carrapatos nesses animais. A abundância de carrapatos encontrados, o hábito da caça, e a criação dos animais de estimação livres em contato com a mata, atuando como hospedeiros-ponte, são fatores que oferecem risco de exposição à Rickettsia spp.. Cabe ressaltar que os dados clínicos de pacientes suspeitos não fornecem informações suficientes para determinar o agente e o vetor envolvidos, principalmente devido ao fato do diagnóstico oficial da febre maculosa se basear unicamente na sorologia do paciente para R. rickettsii. Nosso trabalho reforça a necessidade de que o diagnóstico molecular deveria ser adotado em todos os casos humanos em novas áreas, para determinação da espécie/cepa de Rickettsia spp. circulante em cada região. Considerando as espécies de Rickettsia e seus carrapatos vetores, este trabalho acrescenta um terceiro cenário eco-epidemiológico de febre maculosa para o país: (i) Rickettsia rickettsii associada a Amblyomma sculptum e A. aureolatum; (ii) Rickettsia sp. cepa Mata Atlântica associada a Amblyomma ovale; (iii) R. parkeri associada a A. tigrinum e A. dubitatum. Este trabalho evidencia que a vigilância permanente é necessária para preencher lacunas sobre agentes, reservatórios e vetores de febre maculosa. Em busca do aperfeiçoamento de políticas públicas de vigilância e educação em saúde, é essencial uma melhor compreensão da relação entre natureza, seres humanos e a ocorrência de doenças, particularmente dentro de cada contexto sócio-ambiental.

Spotted fever is an acute febrile disease with nonspecific symptoms. It is caused by bacteria of genus Rickettsia and transmitted by ticks. In this work, we report an eco-epidemiological investigation of two human cases of spotted fever which occurred in Brazilian Pampa biome, in the municipalities of Rosario do Sul in 2011, and Toropi in 2014. In these two areas, ticks were collected from capybaras, dogs and the environment. In Rosário do Sul, blood samples were collected from dogs from the patient domicile and the neighborhood. Also, interviews were carried out with local inhabitants covering questions about ticks and spotted fever. All collected ticks were processed individually to obtain genomic DNA. The search for Rickettsia spp. was carried out by polymerase chain reaction (PCR), which targets the rickettsial fragments of the gltA, ompA and htrA genes. The canine serum samples were also investigated for the presence of antibodies against Rickettsia spp. by indirect immunofluorescence reaction (IFI). In the municipality of Rosário do Sul, the following tick species were found: 251 Amblyomma dubitatum of capybaras carcasses and environment; 60 larvae of Amblyomma spp. from the vegetation; 62 adults of Amblyomma tigrinum, two Amblyomma aureolatum and four Rhipicephalus sanguineus of dogs from patient domicile and neighborhood. Samples of 24 A. tigrinum (38%) and one of Rh. sanguineus were positive in all PCR analyzes (gltA, ompA and htrA) to detect Rickettsia spp. In the municipality of Toropi, 18 A. dubitatum ticks were collected from capybaras carcasses, which four of them were positive for the presence of Rickettsia spp. DNA. Also, 210 A. dubitatum ticks obtained from dead capybara on highways from other locations in the Pampa biome were subjected to PCR analysis, being none of them positive. All positive tick samples (from Rosário do Sul and Toropi) showed 100% identity with Rickettsia parkeri strain Portsmouth. The serology analyses showed that all dogs (100%) have antibodies that reacted with at least one species of Rickettsia, with titres ranging from 128 to 16,384. It was possible to determine R. parkeri as a probable agent responsible for natural infection in 12 samples (66.66%). In the interviews it was observed that the habit of hunting is frequent. All dogs are raised with free contact to forest, and a high rate of parasitism by ticks was observed in these animals. The abundance of ticks, the habit to hunt and to raise the dogs with free contact within the forest (acting as bridge-host) are factors that offer an increased risk of exposure to Rickettsia spp. It is noteworthy, that clinical data of patients do not provide enough information to determine the agent and vector involved, particularly the official diagnosis of spotted fever in Brazil is based solely on the serology of the patient against R. rickettsii. Our research reinforces that molecular diagnosis must be adopted in all human cases of spotted fever to determine which species of Rickettsia circulates in each region. Taking into account the species of Rickettsia and ticks vectors, the set of results showed in this work adds a third eco-epidemiological scenario of spotted fever in Brazil: (i) Rickettsia rickettsii associated with Amblyomma sculptum and A. aureolatum ticks; (ii) Rickettsia sp. strain Atlantic rainforest associated with Amblyomma ovale ticks; (iii) R. parkeri associated with A. tigrinum and A. dubitatum ticks. This work evinces that a permanent survey is necessary to fill the gaps about the agent, reservoirs and vectors of spotted fever. For the improvement of public health surveillances and education policies is necesserary a better understanding about the relationship among nature, human beings and disease ocurrence, particularly considering each socio-enviromental context.

AGENTES DE DOENÇAS TRANSMITIDAS POR VETORES EM CÃES E GATOS DOMÉSTICOS

O presente trabalho relata a ocorrência dos agentes de doenças transmitidas por vetores (VBD) Rangelia vitalii, Babesia vogeli, Hepatozoon spp., Bartonella spp., Anaplasma spp. em cães e gatos domésticos. A presença destes vetores, carrapatos e pulgas, confere a distribuição em larga escala das doenças vetoriais no mundo, que por sua vez, ganharam destaque no ano de 2014 pela Organização Mundial de Saúde (OMS). As doenças transmitidas por vetores podem ser consideradas emergentes ou re-emergentes e, em certas regiões, sua epidemiologia ainda não está definida. Portanto, este estudo objetivou identificar agentes de doenças transmitidas por vetores causadores de hemoparasitoses em cães e gatos domésticos. Amostras de sangue periférico e de ponta de orelha foram colhidas de 110 animais, compreendendo 80 cães e 30 gatos. Os animais eram provenientes dos atendimentos de rotina e de cirurgias de esterilização do Hospital Veterinário da Universidade de Passo Fundo (HV-UPF), cidade de Passo Fundo, região Noroeste do RS. As amostras dos cães foram submetidas a esfregaço sanguíneo coradas com Giemsa e Panótico, detecção de anticorpos anti-Babesia vogeli e anti-Ehrlichia canis através do método de ELISA e PCR para Babesia spp. (gene 18S rRNA), Hepatozoon spp. (gene 18S rRNA), Anaplasma spp. (gene 16S rRNA) e Ehrlichia spp. (gene dsb). As amostras de gatos foram submetidas apenas a esfregaço sanguíneo e PCR para Babesia spp., Hepatozoon spp., Bartonella spp (gene nuoG), Anaplasma spp. e Ehrlichia spp. (gene 16S rRNA). Os esfregaços sanguíneos revelaram um total de 20% (22/110) de amostras positivas, sendo 63,63% (14/22) e 9,09% (2/22) apresentando estruturas semelhantes a mórulas em plaquetas e monócitos, respectivamente. Estruturas compatíveis com piroplasmas foram encontradas em 27,27% (6/22). Os demais agentes não foram identificados por estes métodos. Somente os animais infectados com R. vitalii apresentaram alterações clínicas e hematológicas compatíveis com rangeliose, cujas alterações mais frequentes foram anemia (100%), trombocitopenia (66,7%), icterícia, hemorragias externas e anorexia (50%). Os demais animais positivos identificados com os demais agentes não apresentaram sinais clínicos e/ou alterações hematológicas compatíveis com a infecção causada pelos mesmos. A pesquisa de anticorpos contra B. vogeli mostrou soropositividade de 91% (73/80), enquanto que para B. vogeli e E. canis, apenas 8.75% (7/80). Dezenove (17,27%) dos 110 animais amostrados foram positivos para algum tipo de hemoparasita na PCR. Onze (10%) foram positivos para o gene 18S rRNA de Babesia spp., e após sequenciamento genético identificou-se o agente R. vitalii em 6 (5,5%) dos cães e B. vogeli em 3 cães (2,72%) e 2 gatos (1,81%). Para o gene 18S rRNA de Hepatozoon spp. apenas um cão (0,9%) foi positivo, assim como também apenas um cão (0,9%) foi positivo para o gene 16S rRNA de Anaplasma spp. Para o gene nuoG de Bartonella spp., 6 gatos (20%) foram positivos. Nenhuma amostra foi positiva para os genes dsb e 16S rRNA de Ehrlichia spp. Neste estudo reportamos pela primeira vez no Estado do RS a confirmação molecular de B. vogeli em cães e gatos domésticos. Ademais, os agentes Hepatozoon spp., Anaplasma spp. e Bartonella spp. ainda não haviam sido descritos na região do presente estudo. E, além disso, nós sugerimos que R. vitalii é um dos agentes transmitidos por vetores mais frequentes no Estado do RS.

This study reports the occurrence of vector-borne diseases (VBD) agents Rangelia vitalii, Babesia vogeli, Hepatozoon spp., Anaplasma spp. e Bartonella spp. in domestics dogs and cats. The presence of these vectors, ticks and fleas, gives large-scale distribution of vector-borne diseases in the world, which in turn, have gained prominence in 2014 by the World Health Organization (WHO). The vector-borne diseases can be considered emerging or re-emerging and, in certain regions, its epidemiology is not yet defined. Therefore, this study aimed to identify vector-borne diseases agents by causing hemoparasitosis in domestic dogs and cats. Peripheral blood samples and ear tip were harvested from 110 animals, comprising 80 dogs and 30 cats. The animals were from the routine care and sterilization surgeries Veterinary Hospital of the University of Passo Fundo (HV-UPF), the city of Passo Fundo, Northwest, RS. The samples of dogs underwent blood smear by Giemsa and the Panotic staining, detection of anti-Babesia vogeli antibodies and anti-Ehrlichia canis by ELISA and PCR method for Babesia spp. (18S rRNA gene), Hepatozoon spp. (18S rRNA gene), Anaplasma spp. (16S rRNA) and Ehrlichia spp. (dsb gene). Blood smears revealed a total of 20% (22/110) of positive samples, and 63.63% (14/22) and 9.09% (2/22) having structures similar to morulae platelet and monocytes, respectively. Structures compatible with piroplasms were found in 27.27% (6/22). The other agents were not identified by these methods. Only animals infected with R. vitalii presented clinical and hematological alterations compatible with rangeliose, whose most common were anemia (100%), thrombocytopenia (66.7%), jaundice, external bleeding and anorexia (50%). The other positive animals identified with the other agents did not show clinical signs and/or hematological alterations compatible with infection caused by them. The detection of antibodies against B. vogeli showed seropositivity of 91% (73/80), while for B. vogeli and E. canis, only 8.75% (7/80). Nineteen (17.27%) of the 110 animals sampled were positive for some kind of hemoparasite in PCR. Eleven (10%) were positive for the 18S rRNA gene of Babesia spp., and after genetic sequencing identified the agent R. vitalii in 6 (5.5%) of dogs and B. vogeli in 3 dogs (2.72%) and 2 cats (1.81%). For the 18S rRNA gene Hepatozoon spp. only one dog was positive, as well as only one dogs (0.9%) were positive for 16S rRNA gene of Anaplasma spp. For nuoG gene Bartonella spp., 6 cats (20%) were positive. No sample was positive for dsb and 16S rRNA genes of Ehrlichia spp. In this study we report for the first time in the state of RS the molecular confirmation of B. vogeli in domestic dogs and cats. In addition, agents Hepatozoon spp., Anaplasma spp. and Bartonella spp. had not yet been described in the present study area. Furthermore, we suggest that R. vitalii is one of the most common vector-borne agents in RS.

Interpretation of Coagulation Tendency Contributing to Thrombosis in Vector-Borne Diseases (Ehrlichiosis, Anaplasmosis, Leishmaniosis, and Dirofilariasis) among Dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs.Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid qu

Interpretation of Coagulation Tendency Contributing to Thrombosis in Vector-Borne Diseases (Ehrlichiosis, Anaplasmosis, Leishmaniosis, and Dirofilariasis) among Dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs.Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid qu

Interpretation of Coagulation Tendency Contributing to Thrombosis in Vector-Borne Diseases (Ehrlichiosis, Anaplasmosis, Leishmaniosis, and Dirofilariasis) among Dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs.Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid qu

Interpretation of Coagulation Tendency Contributing to Thrombosis in Vector-Borne Diseases (Ehrlichiosis, Anaplasmosis, Leishmaniosis, and Dirofilariasis) among Dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs.Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid qu

Interpretation of Coagulation Tendency Contributing to Thrombosis in Vector-Borne Diseases (Ehrlichiosis, Anaplasmosis, Leishmaniosis, and Dirofilariasis) among Dogs

Background: Vector-borne infectious and zoonotic diseases are an important health problem that directly affects human and animal health negatively. Results through evaluation of coagulation disorders among vector-borne diseases should be of beneficial for both human and dogs studies. According to the present authors knowledge reports regarding changes in platelet (PLT) count, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer levels in dogs naturally infected with one or more vector-borne pathogens are lacking. Therefore, the present study was aimed to detecting those parameters for relation between diagnosis and prognosis of vector-borne diseases among dogs.Materials, Methods & Results: The material of this study was 46 dogs (36 were naturally infected with vector-borne diseases and 10 were healthy) from different breed, age and of both sexes. Venous blood samples were obtained to detect PLT counts, antibodies of ehrlichiosis, anaplasmosis, borreliosis, leishmaniosis and antigens of Dirofilaria immitis. The diagnosis of vector-borne diseases was performed by using a commercial ELISA assay kits. PLT count was performed with an automated blood cell counter analyser. In addition, PT, APTT and FIB concentrations were measured using a microcoagulometer. D-dimer concentrations were determined using fluorescence immunoassay rapid qu