Thesis (Ph. D.)--Ohio State University, 2008. / Title from first page of PDF file.

La Ehrlichiosis es una zoonosis que ha sido identificada en muchos países de América Latina y el mundo entero. Esta enfermedad ocasiona alteraciones hematológicas y bioquímicas en los pacientes afectados. Los estudios reportados en el Perú muestran valores variables de seropositividad en relación a la enfermedad. En el presente estudio se determinó la presencia de seropositividad a Ehrlichia canis mediante la prueba de Inmunofluorescencia indirecta (IFI) en pacientes humanos y la evaluación hematológica respectiva en cada uno de ellos y su asociación con variables de sexo, edad, exposición a garrapatas y nivel de contacto con perros mediante la prueba de Chi Cuadrado. Se evaluaron 91 sujetos sin distinción de sexo, edad o condición socioeconómica cuyos perros tenían historia de erliqiosis reciente (máximo 6 meses previos al muestreo), los cuales llenaron una encuesta con datos clínico-epidemiológicos de importancia. El muestreo fue realizado entre los meses de febrero y mayo en el Laboratorio de Patología Clínica de la FMV-UNMSM y en los domicilios de cada uno de los pacientes; mientras que, el procesamiento de las muestras se ejecutó en dicho Laboratorio y en la Section de Virología del Laboratorio de Microbiología y Parasitología de dicha facultad. Los resultados de la evaluación hematológica mostraron ausencia de pacientes positivos, habiendo 15.38% de pacientes sospechosos (presencia de corpúsculos de inclusión en células mononucleares y ausencia de trombocitopenia). El porcentaje de pacientes seropositivos por IFI fue de 14.29%, no encontrándose significancia estadística a la prueba de Chi Cuadrado (p menor 0.05) con las variables de interés. Estos hallazgos confirman la exposición a Ehrlichia canis en propietarios de caninos domésticos con antecedentes de Ehrlichiosis en Lima Metropolitana, sin evidenciar asociación de los resultados con los factores de riesgo evaluados. / Ehrlichiosis is a zoonotic disease found in many Latin American countries and worldwide as well. The disease causes hematological and biochemical changes in affected patients. Studies reported in Peru show variable seropositivity values related to this disease. In this study, we evaluated seropositivity and hematologic changes in human subjects by indirect Inmunofluorescent test (IFI) and hematology. We tested the results against risk factors such as sex, age, ticks and dog contact using the Chi-Square test. Ninety-one samples were collected from patients with no difference related to socioeconomic status, age or sex whose dogs had recent history of ehrlichiosis (6 months preview to the sampling) and were asked to fill out a survey with these information. Data recopilation was done between february and may of 2010 at the FMV-UNMSM Clinical Pathology Laboratory and at the subject’s home when necessary. Samples were processed in this laboratory as well as in the Virology Unit of the Microbiology and Parasithology Laboratory. Hematology results revealed an absence of positive cases and 15.38% of suspected cases (showing presence of inclusions bodies within the cytoplasm of mononuclear cells and absence of thrombocytopenia). The IFI seropositive patients were 14.29%, without statistic relevance to the Chi Square test (p menor 0.05) compared to any of the risk factors considered above. These findings confirm exposition to Ehrlichia canis in dog owners with previous record of Ehrilichiosis in Lima Metropolitana, without statistical relationship between tests results and the risk factors evaluated.

La Ehrlichiosis es una zoonosis que ha sido identificada en muchos países de América Latina y el mundo entero. Esta enfermedad ocasiona alteraciones hematológicas y bioquímicas en los pacientes afectados. Los estudios reportados en el Perú muestran valores variables de seropositividad en relación a la enfermedad. En el presente estudio se determinó la presencia de seropositividad a Ehrlichia canis mediante la prueba de Inmunofluorescencia indirecta (IFI) en pacientes humanos y la evaluación hematológica respectiva en cada uno de ellos y su asociación con variables de sexo, edad, exposición a garrapatas y nivel de contacto con perros mediante la prueba de Chi Cuadrado. Se evaluaron 91 sujetos sin distinción de sexo, edad o condición socioeconómica cuyos perros tenían historia de erliqiosis reciente (máximo 6 meses previos al muestreo), los cuales llenaron una encuesta con datos clínico-epidemiológicos de importancia. El muestreo fue realizado entre los meses de febrero y mayo en el Laboratorio de Patología Clínica de la FMV-UNMSM y en los domicilios de cada uno de los pacientes; mientras que, el procesamiento de las muestras se ejecutó en dicho Laboratorio y en la Section de Virología del Laboratorio de Microbiología y Parasitología de dicha facultad. Los resultados de la evaluación hematológica mostraron ausencia de pacientes positivos, habiendo 15.38% de pacientes sospechosos (presencia de corpúsculos de inclusión en células mononucleares y ausencia de trombocitopenia). El porcentaje de pacientes seropositivos por IFI fue de 14.29%, no encontrándose significancia estadística a la prueba de Chi Cuadrado (p menor 0.05) con las variables de interés. Estos hallazgos confirman la exposición a Ehrlichia canis en propietarios de caninos domésticos con antecedentes de Ehrlichiosis en Lima Metropolitana, sin evidenciar asociación de los resultados con los factores de riesgo evaluados. / Ehrlichiosis is a zoonotic disease found in many Latin American countries and worldwide as well. The disease causes hematological and biochemical changes in affected patients. Studies reported in Peru show variable seropositivity values related to this disease. In this study, we evaluated seropositivity and hematologic changes in human subjects by indirect Inmunofluorescent test (IFI) and hematology. We tested the results against risk factors such as sex, age, ticks and dog contact using the Chi-Square test. Ninety-one samples were collected from patients with no difference related to socioeconomic status, age or sex whose dogs had recent history of ehrlichiosis (6 months preview to the sampling) and were asked to fill out a survey with these information. Data recopilation was done between february and may of 2010 at the FMV-UNMSM Clinical Pathology Laboratory and at the subject’s home when necessary. Samples were processed in this laboratory as well as in the Virology Unit of the Microbiology and Parasithology Laboratory. Hematology results revealed an absence of positive cases and 15.38% of suspected cases (showing presence of inclusions bodies within the cytoplasm of mononuclear cells and absence of thrombocytopenia). The IFI seropositive patients were 14.29%, without statistic relevance to the Chi Square test (p menor 0.05) compared to any of the risk factors considered above. These findings confirm exposition to Ehrlichia canis in dog owners with previous record of Ehrilichiosis in Lima Metropolitana, without statistical relationship between tests results and the risk factors evaluated.

ABSTRACT Heartwater, a tick-borne disease caused by Ehrlichia ruminantium, is considered to be a significant cause of mortality amongst domestic and wild ruminants in South Africa. The main vector is Amblyomma hebraeum and although previous epidemiological studies have outlined endemic areas based on mortalities, these have been limited by diagnostic methods which relied mainly on positive brain smears. The indirect fluorescent antibody test (IFA) has a low specificity for heartwater organisms as it cross-reacts with some other species. Since the advent of biotechnology and genomics, molecular epidemiology has evolved using the methodology of traditional epidemiology coupled with the new molecular techniques. A new quantitative real-time polymerase chain reaction (qPCR) test has been developed for rapid and accurate diagnosis of heartwater in the live animal. This method can also be used to survey populations of A. hebraeum ticks for heartwater. Sampling whole blood and ticks for this qPCR differs from routine serumsampling, which is used for many serological tests. Veterinary field staff, particularly animal health technicians, are involved in surveillance and monitoring of controlled and other diseases of animals in South Africa. However, it was found that the sampling of whole blood was not done correctly, probably because it is a new sampling technique specific for new technology, where the heartwater organism is much more labile than the serumantibodies required for other tests. This qPCR technique is highly sensitive and can diagnose heartwater in the living animal within 2 hours, in time to treat it. Poor sampling techniques that decrease the sensitivity of the test will, however, result in a false negative diagnosis. This paper describes the development of a skills training programme for para-veterinary field staff, to facilitate research into the molecular epidemiology of heartwater in ruminants and eliminate any sampling bias due to collection errors. Humane handling techniques were also included in the training, in line with the current focus on improved livestock welfare.

Ehrlichia ruminantium is a tick-borne pathogen transmitted by ticks in the genus Amblyomma. This bacterial pathogen causes heartwater, a fatal disease affecting domestic and wild ruminants in sub-Saharan Africa. The prevalence of heartwater in western Kenya is not well documented. In this study, reverse line blot (RLB) hybridization and quantitative real-time PCR (qPCR) assays were used to detect E. ruminantium DNA in 545 blood samples collected from calves from twenty sublocations distributed across five agro-ecological zones of western Kenya. Ehrlichia ruminantium DNA was detected in 1.10% and 0.92% of the samples using RLB and qPCR, respectively. There were discrepancies in the detection of E. ruminantium by the RLB and the qPCR. Five samples were positive with the qPCR while six were positive with the RLB, but only three of the samples were positive by both tests. The occurrence of E. ruminantium in western Kenya appears to be low, but this might be attributed to the inability of the tests used to detect E. ruminantium carriers. The most prevalent haemoparasites detected by the RLB in the Ehrlichia/Anaplasma group were Anaplasma (formerly Ehrlichia) sp. Omatjenne and Anaplasma bovis at 37.98% each, while Theileria mutans (66.61%) was the most prevalent in the Theileria/Babesia group. In addition, a nested p104 PCR was used to detect Theileria parva in a subset of 86 of the samples; T. parva was detected in 32.56% (28/86) of these samples. The RLB detected T. parva in 27.91% (24/86) of the same sample subset, but only 17 were positive by both tests. The molecular tests used in this study suggest that, of the pathogenic haemoparasites known to cause disease in Kenya, T. parva occurs the most commonly in western Kenya, while E. ruminantium, A. marginale, B. bigemina and B. bovis are less frequently detected. / Dissertation (MSc)--University of Pretoria, 2012. / Veterinary Tropical Diseases / unrestricted

Para elucidar alguns aspectos da imunopatogenia da uveíte na erliquiose canina, avaliaram-se, por meio da análise imunistoquímica, bulbos oculares de cães experimentalmente infectados por Ehrlichia canis (grupo 1- G1), naturalmente infectados por Ehrlichia canis (grupo 2-G2) e na co-infecção natural de E. canis e Babesia sp. (grupo 3). Parâmetros clínicos e hematológicos foram avaliados. Empregaram-se o dot-blot-Elisa e a reação de imunofluorescência indireta para E. canis e Babesia sp. respectivamente. Para a confirmação diagnóstica, utilizou-se a reação de cadeia de polimerase (PCR) para E.canis. A contagem imunofenotípica para os anticorpos CD3, CD4, CD8, Tal1B5 e MAC 387 não demonstrou diferença significativa nas diferentes regiões analisadas do bulbo ocular. Observou-se no G1, G2 e G3, em todas as regiões analisadas diferença significativa da contagem imunofenotípica de células CD8+ em relação às células CD4+. Evidenciou-se diferença significativa entre a contagem percentual de células IgG2+ e CD79?+ na região de corpo ciliar do G3 em relação ao G1. A região da íris do G3, em relação ao G2, demonstrou diferenças significativas para o anticorpo IgG1. Evidenciou-se nos três grupos, a existência de correlação linear entre as células CD3+ e CD8+ e entre as células IgG2+ e CD79?+ em diversas regiões do bulbo ocular. O infiltrado inflamatório mostrou-se mais intenso nas regiões de corpo ciliar e ângulo iridocorneal, moderado em limbo e íris e mínimo em coróide. A avaliação semiquantitativa por score da intensidade do infiltrado inflamatório mostrou-se mais intensa nos animais que apresentavam co-infecção, sugerindo uma resposta imune mais intensa nesses cães. Demonstrou-se que o infiltrado inflamatório era composto, predominantemente, por linfócitos T CD3+ e B CD79+?. A maior porcentagem de células T CD3+ era CD8+, caracterizando, portanto, uma resposta imune do tipo citotóxica. A presença de células B CD79+? fala a favor de produção local de anticorpos. Observaram-se células imunomarcadas por IgG2 e poucas células marcadas por IgG1, sugerindo uma polarização da resposta imune para o padrão Th1, sendo um possível mecanismo imune de lesão na uveíte. Observou-se alta expressão de moléculas de MHC-classe II, sugerindo uma resposta imune intensa nos tecidos oculares, contudo ineficiente devido a deficiência de células CD4+. / The immunopathogenicity of uveitis in the canine ehrlichiosis was studied by conducting anatomy and immunohistochemical analyses in the ocular globes of dogs experimentally (Group 1) and naturally (Group 2) infected with Ehrlichia canis, and naturally coinfected with Ehrlichia canis and Babesia sp. (Group 3). Clinical and hematological parameters were evaluated. Dot-blot Elisa and indirect immunofluorescent test (IFA) were used to analyze E. canis and Babesia sp., respectively. PCR assay confirmed the diagnosis of the disease caused by E. canis. The immunophenotypic analysis with the antibodies CD3, CD4, CD8, Tal1B5 and MAC 387 revealed no significant differences between the various ocular regions analyzed. Significant differences were observed between the immunophenotypic analysis of CD8+ and CD4+cells from all regions analyzed from G1, G2 and G3; between the percentile counts of IgG2+ and CD79?+ in the ciliary body of G3 dogs when compared to G1, and for the IgG1 antibody counts of the iris region from G3 when compared to G2. A linear correlation between CD3+ and CD8+ cells and between IgG2+ and CD79?+ cells from several regions of the ocular globe was found for all three groups. The cellular inflammatory infiltrate observed in the ocular tissue was severe in the regions of the ciliary bodies and iridocorneal angle, moderate in the limbus and iris, and only slightly present in the choroids. A semiquantitative analysis of the intensity of the inflammatory infiltrate of the ocular globes was more intense in G3, which suggested a greater response of the immune system in these animals. The inflammatory infiltrate was composed of mainly B CD79+? and T CD3+ lymphocytes, which had CD8+ at a high percentage, thus characterizing this as a cytotoxic immune response. Results indicated that B CD79+? cells favored the production of local antibodies. IgG2 and very few IgG1 immunolabeled cells were found. This result indicated a polarization of the immune response to the Th1 pattern, which could be the mechanism of the lesions in the uveitis. A large number of cells expressing the MHC-class II molecules were observed, suggesting an intense immune response in the ocular tissues, however ineffective due to the CD4+ cell´s deficient.

The aim of this study was to determine the serologic and molecular occurrence of Rickettsia spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. in ticks and domestic and wild animals from two conservation units in the city of Natal, Rio Grande do Norte State, Brazil. The collection period was between October 2012 and August 2013. Serum samples were tested against Rickettsia spp. antigens and Ehrlichia canis by Indirect fluorescent antibody test. Tissue samples and ticks were processed for molecular detection of the pathogens. Twenty-seven marsupials and four rodents were captured, and up to three animals of each species were euthanized. In addition, serum samples from 155 domestic animals: 53 cats living inside the units, 29 dogs domiciled around the areas and 73 dgos of the Zoonosis Control Center of the City (ZCC). Twenty dogs from ZCC were also euthanized and samples of spleen were obtained. Antibodies to at least one of the Rickettsia species tested were detected in six Didelphis albiventris and in one Rattus rattus; 17% (17/102) of the dogs presented antibodies to E. canis and 13% (20/155) of all tested domestic animals (dogs and cats) were seropositive for Rickettsia spp. antigens. Three species of ticks (Amblyomma auricularium, Ixodes loricatus and Ornithodoros mimon) were collected and one A. auricularium was positive for Rickettsia amblyommii by PCR. Two D. albiventris spleen samples amplified PCR products for Ehrlichia spp. Spleen samples from three D. albiventris and spleen and lung sample from one Necromys lasiurus were positive for Babesia spp. by PCR test. Among the 20 spleen samples from dogs subjected to molecular analysis, eight were positive by PCR for E. canis and two for H. canis / O objetivo deste estudo foi determinar a ocorrência sorológica e molecular de Rickettsia , Ehrlichia , Hepatozoon e Babesia em carrapatos e mamíferos silvestres e domésticos, provenientes de duas unidades de conservação ambiental (UC) na cidade de Natal, Rio Grande do Norte, Brasil. O período de coleta foi de outubro de 2012 a agosto de 2013. Os soros foram testados contra antígenos de Rickettsia spp. e Ehrlichia canis através da reação de imunofluorescência indireta. Amostras de tecido e carrapatos foram processadas para a detecção molecular dos patogenos. Foram capturados 27 marsupiais e quatro roedores para coleta de sangue, destes foram eutanásiados ate três animais de cada espécie e coletadas amostras de baço e pulmão. Paralelo, amostras de soro de 155 animais domésticos: 53 gatos que viviam nas UCs, 29 cães domiciliados no entorno das areas e 73 cães do Centro de Controle de Zoonoses do município, dos quais 20 tiveram amostras de baço coletadas. Foram detectados anticorpos para, pelo menos, uma das espécies de Rickettsia testadas em seis Didelphis albiventris e em um Rattus rattus , e 17 % (17/102) dos cães apresentaram anticorpos anti- E. canis e 13% (20/155) de todos os animais domésticos (cães e gatos) foram soropositivos para antígenos de Rickettsia spp. Três espécies de carrapatos (Amblyomma auricularium , Ixodes loricatus e Ornithodoros mimon ) foram coletadas e um A. auricularium foi positivo para R. amblyommii pela PCR. Duas amostras de baço de D. albiventris amplificaram produtos de PCR para Ehrlichia spp. e amostras de baço de três D. albiventris e baço e pulmão de um Necromys lasiurus foram positivas para Babesia spp. pela PCR. Entre as 20 amostras de baço de cão submetidas a análises moleculares, oito foram positivas na PCR para E. canis e duas para H. canis

Para elucidar alguns aspectos da imunopatogenia da uveíte na erliquiose canina, avaliaram-se, por meio da análise imunistoquímica, bulbos oculares de cães experimentalmente infectados por Ehrlichia canis (grupo 1- G1), naturalmente infectados por Ehrlichia canis (grupo 2-G2) e na co-infecção natural de E. canis e Babesia sp. (grupo 3). Parâmetros clínicos e hematológicos foram avaliados. Empregaram-se o dot-blot-Elisa e a reação de imunofluorescência indireta para E. canis e Babesia sp. respectivamente. Para a confirmação diagnóstica, utilizou-se a reação de cadeia de polimerase (PCR) para E.canis. A contagem imunofenotípica para os anticorpos CD3, CD4, CD8, Tal1B5 e MAC 387 não demonstrou diferença significativa nas diferentes regiões analisadas do bulbo ocular. Observou-se no G1, G2 e G3, em todas as regiões analisadas diferença significativa da contagem imunofenotípica de células CD8+ em relação às células CD4+. Evidenciou-se diferença significativa entre a contagem percentual de células IgG2+ e CD79?+ na região de corpo ciliar do G3 em relação ao G1. A região da íris do G3, em relação ao G2, demonstrou diferenças significativas para o anticorpo IgG1. Evidenciou-se nos três grupos, a existência de correlação linear entre as células CD3+ e CD8+ e entre as células IgG2+ e CD79?+ em diversas regiões do bulbo ocular. O infiltrado inflamatório mostrou-se mais intenso nas regiões de corpo ciliar e ângulo iridocorneal, moderado em limbo e íris e mínimo em coróide. A avaliação semiquantitativa por score da intensidade do infiltrado inflamatório mostrou-se mais intensa nos animais que apresentavam co-infecção, sugerindo uma resposta imune mais intensa nesses cães. Demonstrou-se que o infiltrado inflamatório era composto, predominantemente, por linfócitos T CD3+ e B CD79+?. A maior porcentagem de células T CD3+ era CD8+, caracterizando, portanto, uma resposta imune do tipo citotóxica. A presença de células B CD79+? fala a favor de produção local de anticorpos. Observaram-se células imunomarcadas por IgG2 e poucas células marcadas por IgG1, sugerindo uma polarização da resposta imune para o padrão Th1, sendo um possível mecanismo imune de lesão na uveíte. Observou-se alta expressão de moléculas de MHC-classe II, sugerindo uma resposta imune intensa nos tecidos oculares, contudo ineficiente devido a deficiência de células CD4+. / The immunopathogenicity of uveitis in the canine ehrlichiosis was studied by conducting anatomy and immunohistochemical analyses in the ocular globes of dogs experimentally (Group 1) and naturally (Group 2) infected with Ehrlichia canis, and naturally coinfected with Ehrlichia canis and Babesia sp. (Group 3). Clinical and hematological parameters were evaluated. Dot-blot Elisa and indirect immunofluorescent test (IFA) were used to analyze E. canis and Babesia sp., respectively. PCR assay confirmed the diagnosis of the disease caused by E. canis. The immunophenotypic analysis with the antibodies CD3, CD4, CD8, Tal1B5 and MAC 387 revealed no significant differences between the various ocular regions analyzed. Significant differences were observed between the immunophenotypic analysis of CD8+ and CD4+cells from all regions analyzed from G1, G2 and G3; between the percentile counts of IgG2+ and CD79?+ in the ciliary body of G3 dogs when compared to G1, and for the IgG1 antibody counts of the iris region from G3 when compared to G2. A linear correlation between CD3+ and CD8+ cells and between IgG2+ and CD79?+ cells from several regions of the ocular globe was found for all three groups. The cellular inflammatory infiltrate observed in the ocular tissue was severe in the regions of the ciliary bodies and iridocorneal angle, moderate in the limbus and iris, and only slightly present in the choroids. A semiquantitative analysis of the intensity of the inflammatory infiltrate of the ocular globes was more intense in G3, which suggested a greater response of the immune system in these animals. The inflammatory infiltrate was composed of mainly B CD79+? and T CD3+ lymphocytes, which had CD8+ at a high percentage, thus characterizing this as a cytotoxic immune response. Results indicated that B CD79+? cells favored the production of local antibodies. IgG2 and very few IgG1 immunolabeled cells were found. This result indicated a polarization of the immune response to the Th1 pattern, which could be the mechanism of the lesions in the uveitis. A large number of cells expressing the MHC-class II molecules were observed, suggesting an intense immune response in the ocular tissues, however ineffective due to the CD4+ cell´s deficient.

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Salvador, BA, Brasil / A Erliquiose Monocítica Canina (EMC) e a Leishmaniose Visceral Canina (LVC) são duas doenças, transmitidas por vetores, com ampla distribuição mundial. Os agentes causadores dessas doenças são Ehrlichia canis e Leishmania infantum, respectivamente. Enquanto a EMC é transmitida por carrapatos, principalmente Rhipicephalus sanguineus, a Leishmania é inoculada no hospedeiro através do inseto-vetor da subfamilia Phlebotominae. A maioria dos sinais físicos são comuns às duas enfermidades, o que dificulta o diagnóstico clínico e o tratamento, especialmente em áreas endêmicas. Poucos são os estudos relacionados à coinfecção da erliquiose e leishmaniose caninas no Brasil. No Piauí, particularmente em Teresina, ainda não se tem dados concretos sobre a prevalência dessas afecções.Os sinais físicos variam com a severidade da infecção, a resposta imune do hospedeiro e a presença de coinfecção. Assim, buscamos avaliar cães, independente dos sinais físicos, atendidos no hospital universitário e clínicas particulares da cidade de Teresina, Piauí quanto a frequência de erliquiose e leishmaniose caninas no período de março de 2012 a setembro de 2014, independente de sexo, raça, idade e suspeita clínica, excetuando os cães que vinham apenas para vacinação. Foi realizado exame clínico e colhidas amostras sanguíneas para exame hematimétrico, bioquímico, parasitológico e molecular. Dos 664 cães avaliados para E. canis, 22,9% tiveram o DNA amplificado somente para esta bactéria. No caso de Leishmania infantum, houve amplificação de DNA em 29,1% dos 664 cães, sendo que, destes, 80 (12,1%) estavam coinfectados com E.canis. Anemia e trombocitopenia foram as principais alterações hematológicas observadas em cães infectados tanto com E. canis, como com L. infantum e coinfectados. As principais alterações bioquímicas nos cães com infecção apenas por L. infantum, assim como nos coinfectados, foram hipoglobulinemia e hiperproteinemia. Já nos monoinfectados por E. canis a hipoalbuminemia e a uremia foram as principais alterações. Perda de peso, úlceras, apatia, conjuntivite, palidez nas mucosas, alópecia e linfoadenomegalia são mais frequentes em cães monoinfectados com L. infantum do que nos infectados apenas com E. canis. Além disso, os cães coinfectados apresentam uma doença mais severa do que os monoinfectados. Concluímos que a presença da coinfecção por esses agentes infecciosos é endêmica na cidade de Teresina-Piauí e que existe um sinergismo entra as duas doenças.Portanto, destacamos aqui a relevância dos estudos voltados a epidemiologia e melhoria dos testes diagnósticos para uma prevenção e/ou controle mais eficazes dessas doenças. Palavras / The Canine Monocytic Ehrlichiosis (CME) and Canine Visceral Leishmaniasis (CVL) are the two vector-borne diseases with worldwide distribution. The causative agents of these diseases are Ehrlichia canis and Leishmania infantum, respectively. While CME is transmitted by ticks, especially Rhipicephalus sanguineus, Leishmania is inoculated into the host through the insect vector of the subfamily Phlebotominae. Most clinical signs are common to both conditions, which hinders the clinical diagnosis and treatment, especially in endemic areas. There are few studies related to the coinfection of canine ehrlichiosis and leishmaniasis in Brazil. In Piaui, particularly in Teresina, there is not yet concrete data on the prevalence of these diseases. The clinical signs vary according to the severity of the infection, to the host immune response and to the presence of coinfection. Thus, we sought to evaluate dogs, regardless of clinical signs, seen at university hospital and private clinics in the city of Teresina, Piauí, for the frequency of canine ehrlichiosis and leishmaniasis from March 2012 to September 2014, regardless of sex, race, age and clinical suspicion, except dogs that came just to vaccination purposes. Clinical examination was performed and blood samples for erythrocyte examination, biochemical, and molecular parasitology were collected. From 664 dogs evaluated for E. canis, 22, 9% showed amplified DNA, to this bacterium only . Regarding to Leishmania infantum, there was DNA amplification in 29, 1% out of the 664 dogs, and, from these, 80 (12, 1%) were coinfected with E. canis. Anemia and thrombocytopenia were major hematologic changes observed in dogs infected with Leishmania, E.canis and coinfected dogs. The main biochemical findings in dogs infected by L. infantum and coinfected were hyperalbuminemia, hypoglobulinemia and hyperproteinemia. In those infected only by E. canis, hypoalbuminemia and uremia were the main changes. Weight loss, skin lesions, lethargy, conjunctivitis, pale mucous membranes, alopecia and enlarged lymphnodes are more common in dogs with L. infantum than in dogs infected only with E.canis. In addition, the coinfected dogs have a more severe disease than the ones with only one infection. We conclude that the presence of coinfection by these infectious agents are endemic in the city of Teresina, Piauí, and that there is a synergism between the two diseases. Therefore, we highlight here the relevance of studies regarding epidemiology and improvement of diagnostic tests for prevention and / or control of the diseases in a more effective way.

The aim of this study was to determine the serologic and molecular occurrence of Rickettsia spp., Ehrlichia spp., Babesia spp. and Hepatozoon spp. in ticks and domestic and wild animals from two conservation units in the city of Natal, Rio Grande do Norte State, Brazil. The collection period was between October 2012 and August 2013. Serum samples were tested against Rickettsia spp. antigens and Ehrlichia canis by Indirect fluorescent antibody test. Tissue samples and ticks were processed for molecular detection of the pathogens. Twenty-seven marsupials and four rodents were captured, and up to three animals of each species were euthanized. In addition, serum samples from 155 domestic animals: 53 cats living inside the units, 29 dogs domiciled around the areas and 73 dgos of the Zoonosis Control Center of the City (ZCC). Twenty dogs from ZCC were also euthanized and samples of spleen were obtained. Antibodies to at least one of the Rickettsia species tested were detected in six Didelphis albiventris and in one Rattus rattus; 17% (17/102) of the dogs presented antibodies to E. canis and 13% (20/155) of all tested domestic animals (dogs and cats) were seropositive for Rickettsia spp. antigens. Three species of ticks (Amblyomma auricularium, Ixodes loricatus and Ornithodoros mimon) were collected and one A. auricularium was positive for Rickettsia amblyommii by PCR. Two D. albiventris spleen samples amplified PCR products for Ehrlichia spp. Spleen samples from three D. albiventris and spleen and lung sample from one Necromys lasiurus were positive for Babesia spp. by PCR test. Among the 20 spleen samples from dogs subjected to molecular analysis, eight were positive by PCR for E. canis and two for H. canis / O objetivo deste estudo foi determinar a ocorrência sorológica e molecular de Rickettsia , Ehrlichia , Hepatozoon e Babesia em carrapatos e mamíferos silvestres e domésticos, provenientes de duas unidades de conservação ambiental (UC) na cidade de Natal, Rio Grande do Norte, Brasil. O período de coleta foi de outubro de 2012 a agosto de 2013. Os soros foram testados contra antígenos de Rickettsia spp. e Ehrlichia canis através da reação de imunofluorescência indireta. Amostras de tecido e carrapatos foram processadas para a detecção molecular dos patogenos. Foram capturados 27 marsupiais e quatro roedores para coleta de sangue, destes foram eutanásiados ate três animais de cada espécie e coletadas amostras de baço e pulmão. Paralelo, amostras de soro de 155 animais domésticos: 53 gatos que viviam nas UCs, 29 cães domiciliados no entorno das areas e 73 cães do Centro de Controle de Zoonoses do município, dos quais 20 tiveram amostras de baço coletadas. Foram detectados anticorpos para, pelo menos, uma das espécies de Rickettsia testadas em seis Didelphis albiventris e em um Rattus rattus , e 17 % (17/102) dos cães apresentaram anticorpos anti- E. canis e 13% (20/155) de todos os animais domésticos (cães e gatos) foram soropositivos para antígenos de Rickettsia spp. Três espécies de carrapatos (Amblyomma auricularium , Ixodes loricatus e Ornithodoros mimon ) foram coletadas e um A. auricularium foi positivo para R. amblyommii pela PCR. Duas amostras de baço de D. albiventris amplificaram produtos de PCR para Ehrlichia spp. e amostras de baço de três D. albiventris e baço e pulmão de um Necromys lasiurus foram positivas para Babesia spp. pela PCR. Entre as 20 amostras de baço de cão submetidas a análises moleculares, oito foram positivas na PCR para E. canis e duas para H. canis