Calves infected with Babesia bovis or Babesia bigemina between 3 and 9 months of age can develop immunity without showing overt clinical signs. This transient immunity is not dependent on maternal immunity. After 9 months of age, they are fully susceptible to challenge. Dairy calves between 2 and 3 months of age were vaccinated with B. bigemina and B. bovis live frozen vaccines (Onderstepoort Biological Products®). Two months after vaccination, 90% of calves were serologically positive on IFA test to B. bigemina, and 70% were serologically positive to B. bovis. At this age, only 17% of the control group had seroconverted to B. bigemina and none of the calves had seroconverted to B. bovis. All experimental calves maintained positive serological status to both B. bovis</i. and B. bigemina for at least 5 months after vaccination. It is sound practice to vaccinate dairy calves against babesiosis at 2–3 months of age. Endemic stability is achieved before the period of natural resistance wanes. Copyright / Dissertation (MSc)--University of Pretoria, 2011. / Veterinary Tropical Diseases / unrestricted

O carrapato Rhipicephalus microplus é um ectoparasito hematófago de grande importância para a pecuária por ser responsável por perdas massivas na produção animal, de forma que o seu controle é economicamente relevante. Este carrapato, além dos danos que causa por si só, é também um importante vetor para a transmissão de microorganismos patogênicos, entre eles o hemoprotozoário intraeritrocítico Babesia bovis. O presente trabalho descreve o desenvolvimento de uma linhagem de B. bovis capaz de expressar um antígeno protetor, uma glutationa S-transferase do carrapato Haemaphysalis longicornis (HlGST), e o teste desta linhagem como uma vacina de vetor vivo para o controle do carrapato R. microplus. B. bovis, em cultivo, da linhagem S74-T3B foram eletroporados em presença de plasmídeo contendo o promotor bidirecional de B. bovis Ef-1 aresponsável pela expressão independente de dois genes: o repórter fusionado ao agente para seleção (GFP-BSD) e HlGST fusionada à sequência codificadora do peptídeo sinal de MSA-1 (merozoite surface antigen-1). Após a eletroporação, foi feita a seleção com blasticidina para obtenção da linhagem nomeada HlGST. A linhagem HlGST é composta por parasitos contendo diferentes padrões de inserção dos genes exógenos, tanto dentro quanto fora do locus Ef-1. Uma linhagem clonal denominada HlGST-Cln expressando HlGST e GFP-BSD foi obtida a partir da linhagem HlGST. Dois ensaios, independentes, de imunização de bovinos com os parasitos clonais foram realizados, sendo usado como controle uma linhagem clonal previamente caracterizada denominada GFP-Cln. Todos os animais inoculados desenvolveram uma forma branda de babesiose, indicando que ambas as linhagens clonais são atenuadas, mas apenas os animais imunizados com a linhagem HlGST-Cln foram capazes de produzir anticorpos anti-HlGST. O segundo procedimento de imunização foi seguido por um desafio com larvas de R. microplus. O desenvolvimento dessas larvas no hospedeiro levou a fêmeas adultas de menor peso e fertilidade. Coletivamente, esses dados mostram a possibilidade de uso de linhagens transfectadas de B. bovis como vacinas de vetor vivo. / The tick Rhipicephalus microplus is a notorious blood-feeding ectoparasite of cattle, responsible for massive losses in animal production. It is the main vector of pathogenic microorganisms, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74- T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1 promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD, and HlGST fused to the MSA-1 (merozoite surface antigen-1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1 a locus. A B. bovis clonal line termed HlGST-Cln expressing HlGST and GFP-BSD was then derived from the mixed parasite line HlGST. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1 locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis indicating that both transfected cloned parasite lines are attenuated. All animals immunized with HlGST-Cln produced detectable anti-glutathione-S-transferase antibodies. After immunization with HlGST-Cln, calves were challenged with R. microplus larva. Development of these larva produce fully engorged female tick with reduced weight and fertility. Collectively, these data show that transfected B. bovis parasites can be used as vectors in live vectored vaccines.

Canine tick-borne diseases are an emerging problem within Australia and throughout the world. This thesis investigates Babesia gibsoni and Anaplasma platys infections in dogs in Australia and also explores the evolutionary relationships and taxonomy of the canine piroplasm species and the members of the order Piroplasmida. A nested PCR-RFLP assay was developed for the detection and differentiation of the canine piroplasm species and was found to have a high detection limit, capable of detecting a 2.7 x 10-7 % parasitaemia or the equivalent of 1.2 molecules of target DNA. Detection of piroplasm DNA applied to Whatman FTA“ classic cards using nested-PCR was found to have a lower detection limit than when using DNA extracted from whole blood but higher than IsoCode‘ Stix or QIAamp extraction from filter paper based techniques. The nested PCR-RFLP assay was further evaluated for the detection of B. gibsoni infection in dogs being exported from Australia to New Zealand and compared to the current screening methods, the Immunofluorescent Antibody Test (IFAT) and microscopy. Of 235 dogs screened, 11 were IFAT positive, 1 was microscopy positive and 3 were PCR positive for B. gibsoni, highlighting the discordance that exists between various detection techniques. Replacing microscopic examination of blood smears with PCR-RFLP is suggested for screening dogs entering New Zealand, in addition to revising the current IFAT cut-off titre to minimize false positive results. The first case of B. gibsoni in New South Wales is also reported. A study was also conducted to further investigate the recent discovery of B. gibsoni in Australia and the association of this infection with American Pit Bull Terriers in an epidemiological study. Both American Pit Bull Terriers (n = 100) and other dog breeds (n = 51) were screened for B. gibsoni using IFAT and PCR-RFLP. A questionnaire was also completed by each dog owner regarding thethe husbandry and habits these dogs. Fourteen dogs were positive for B. gibsoni using IFAT and/or PCR-RFLP and all were American Pit Bull Terriers. Dogs that were male and/or were bitten by or were biters of other American Pit Bull Terriers were statistically more likely to be B. gibsoni positive, thus suggesting that blood-to-blood transmission may contribute to the spread of this disease. Experimental B. gibsoni infections were established in vivo to investigate the efficacy of combined atovaquone and azithromycin therapy and to determine the detection limits of PCR, IFAT and microscopy during various stages of infection. While atovaquone and azithromycin produced a reduction in circulating parasite levels, it did not cause total eradication, and possible drug resistance also developed in one dog. PCR was found to be most useful in detecting early and acute stage infections, while IFAT was most useful during chronic and acute infections. Microscopy is suggested to be only useful for detecting acute stage infections. This study also describes the detection of B. gibsoni in tissue samples during chronic infection for the first time, suggesting possible sequestration of this parasite. Anaplasma platys has also only recently been reported in Australia and the distribution, molecular-charcterisation, pathogenesis, co-infection with Babesia canis vogeli and treatment of infection with doxycycline were investigated. For the first time, A. platys is reported in Western Australia, Queensland and Victoria, with each isolate found to be genetically identical on the basis of the 16S rRNA gene. No correlation could be established between A. platys infection and the development of clinical signs or pathogenesis and definitive treatment using doxycycline could not be determined. Isolates of canine piroplasms from various geographical locations worldwide (n = 46), including Australia were characterised on the basis of multiple gene loci to explore the distribution, genetic variation and possible phylogeographical relationships of these species. Separate genotypes of B. canis vogeli, B. canis canis and B. gibsoni are suggested and may be correlated to different geographical origins. Characterization of B. canis vogeli, B. canis canis and B. canis rossi on the basis of the HSP 70 gene and B. gibsoni on the basis of the ITS 1, 5.8S rRNA gene and ITS 2 is described for the first time. Elevation of each of the B. canis subspecies, with the exclusion of B. canis presentii, to separate species is also proposed. The current paraphyly and taxonomic confusion associated with the members of the order Piroplasmida led to a review of the phylogenetic and taxonomic status of this group of organisms. Phylogenetic relationships are determined using 18S rRNA gene, 5.8S rRNA gene, HSP 70 gene and combined loci analyses. Rearrangement of the Piroplasmida into three families, including the new family Piroplasmiidae is proposed, in addition to the establishment of two new genera, the Piroplasma (Patton, 1895) and the Achromaticus (Dionisi, 1899). Other proposed schemes of classification and the limitations of phenotypic characteristics in taxonomic classification within the Piroplasmida are also discussed.

Thesis (MSc (Veterinary Science))--University of Pretoria, 2001. / Includes bibliographical references.

Thesis (MSc (Veterinary Science))--University of Pretoria, 2005. / Includes bibliographical references.

Thesis (M.A.) -- Central Connecticut State University, 2006. / Thesis advisor: Kathy Martin. "... in partial fulfillment of the requirements for the degree of Master of Arts in Biomolecular Sciences." Includes bibliographical references (leaves 99-103). Also available via the World Wide Web.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-06-07T18:34:57Z No. of bitstreams: 1 texto completo.pdf: 620244 bytes, checksum: fb04690385958b310ed3e970511278f3 (MD5) / Made available in DSpace on 2017-06-07T18:34:57Z (GMT). No. of bitstreams: 1 texto completo.pdf: 620244 bytes, checksum: fb04690385958b310ed3e970511278f3 (MD5) Previous issue date: 2001-03-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Eritrócitos de bovinos inoculados com amostras de Babesia bovis (BbovUFV1 7 a passagem e Jaboticabal 7 a passagem, patogênicas e BbovUFV1 26 a passagem, atenuada) foram testados quanto à sua capacidade de aderir em células endoteliais de aorta bovina (BAECs) “in vitro”. Paralelamente, foram realizados ensaios de imunofluorescência indireta com amostra de sangue periférico dos animais para detecção da presença de eritrócitos portando antígenos de B. bovis em sua superfície e testes de hemaglutinação passiva para detecção de antígenos livres de B. bovis nos soros dos mesmos. Houve aumento significativo no número de eritrócitos não parasitados aderidos dos animais inoculados com amostra patogênica BbovUFV1 7 a passagem, que se associou ao aparecimento de antígenos livres de B. bovis no soro e de eritrócitos não parasitados marcados com antígenos do parasita no sangue periférico. Estes resultados não foram observados nos testes com amostras dos animais inoculados com BbovUFV1 26 a passagem atenuada. Eritrócitos obtidos dos animais inoculados com amostra patogênica Jaboticabal 7 a passagem não se mostraram aderentes às BAECs a despeito dos animais terem apresentado eritrócitos não parasitados modificados com antígenos de B. bovis em sua superfície e níveis transitórios de antígenos livres no soro. Os dados sugerem que a modificação da superfície de eritrócitos não parasitados por antígenos de B. bovis presentes no soro dos animais inoculados pode levar a adesão eritrocitária “in vitro”. Porém as modificações não culminam sempre em desenvolvimento de adesão, visto que os antígenos de B. bovis que marcam os eritrócitos não parasitados podem variar em sua estrutura e capacidade de ligação em receptores endoteliais. / Erythrocytes from cattle infected with Babesia bovis strains BbovUFV1 7 th passage and Jaboticabal 7 th passage, both pathogenic, and BbovUFV1 26 th passage, attenuated strain, were assayed for binding to bovine aortic endothelial cells (BAECs) “in vitro”. In parallel, periferal blood samples were assayed in an indirect immunofluorescense antibody test (IFAT) to detect erythrocytes bearing B. bovis antigens on its surface and serum samples were used in haemagglutination tests for detection of free soluble B. bovis antigens. A significant increase in the number of adherent erythrocytes was obseved for erythrocytes from animals infected with the pathogenic strain BbovUFV1 7 th passage, which was associated to the appearance of B. bovis antigens released in the serum, as well as to the appearance of non-parasitised erythrocytes marked with parasite’s antigens. These results were not observed in tests with serum and blood samples of animals inoculated with the attenuated strain BbovUFV1 26 th passage. Erythrocytes obtained from animals inocculated with pathogenic strain Jaboticabal 7 th passage did not adhere to BAECs, despite the presence of non-parasitized erythrocytes modified with B. bovis in blood and transient levels of free antigens in serum of inocculated animals. The data suggests that surface modifications on non-parasitized erythrocytes by B. bovis antigens in the serum of the inocculated animals could lead to erythrocytic adhesion “in vitro”. However, these modifications do not always lead to adhesion, because the B. bovis antigens which mark the non-parasitised erythrocytes can vary in structure and endothelial receptors binding properties. / Dissertação importada do Alexandria

Submitted by Nathália Faria da Silva (nathaliafsilva.ufv@gmail.com) on 2017-06-07T19:51:19Z No. of bitstreams: 1 texto completo.pdf: 2564154 bytes, checksum: ee2fe00d1c7a56d422a883011419d135 (MD5) / Made available in DSpace on 2017-06-07T19:51:19Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2564154 bytes, checksum: ee2fe00d1c7a56d422a883011419d135 (MD5) Previous issue date: 2002-09-09 / Fundação de Amparo a Pesquisa do Estado de Minas Gerais / Foram isoladas células endoteliais de veia umbilical bovina (BUVECs) com a finalidade de determinar a expressão de moléculas de adesão como ICAM-1, VCAM, PECAM-1, E-selectina e trombospondina na fisiopatologia de B. bovis num estudo in vitro. Posteriormente esta expressão foi confirmada, num estudo in situ, em tecidos (cérebro, pulmão e rim) de animais que morreram após inoculação com amostra patogênica de Babesia bovis (BbovUFV1 7 a passagem). Eritrócitos dos animais infectados foram testados quanto à capacidade de aderir em BUVECs determinando a cinética de adesão. Os mesmos testes de adesão foram realizados em BUVECs estimuladas com plasma de animais infectados com B. bovis e sobrenadante de cultura de PBMC bovino estimulado com peptídeo sintético proveniente da RAP-1 de B. bovis contendo citocinas quantificadas (IFN-γ, TNF-α e IL-12). Houve aumento significativo na adesão de eritrócitos de animais inoculados em BUVECs estimuladas com plasma e sobrenadante de PBMC. Entretanto, adesão foi observada somente para eritrócitos não parasitados, sugerindo que antígenos livres de B. bovis no soro marcam eritrócitos não parasitados, ou ainda uma possível expressão de uma isoforma de VESA-1 não aderente. Aderência não foi observada nos testes com amostras dos animais negativos. As células estimuladas com plasma de animais infectados e com sobrenadante de PBMC mostraram imunomarcação positiva muito mais intensa de ICAM-1, VCAM, PECAM-1, E-selectina e trombospondina que as células que não receberam estímulo. Igualmente no estudo in situ foi observado aumento na expressão de ICAM- 1, VCAM, PECAM-1, E-selectina e trombospondina em tecidos como cérebro, pulmão e rim dos bovinos infectados com B. bovis em comparação ao grupo controle. Estes resultados sugerem que as interleucinas, liberadas na fase aguda da babesiose, estimulam a expressão das moléculas de adesão relacionadas à fisiopatologia da babesiose causada por B. bovis, fato demonstrado in vitro pela expressão das moléculas em BUVECs e a citoaderência de eritrócitos. Estes achados demonstram similaridades na fisiopatologia de B. bovis e do Plasmodium falciparum. / Endothelial cells from bovine umbilical vein (BUVECs) were isolated with the purpose of determining the expression of ICAM-1, VCAM, PECAM- 1, E-selectin and thrombospondin in the physiopathology of Babesiosis caused by B. bovis. Later, this expression was confirmed by in situ study, using tissue samples (brain, lung and kidney) of animals that died after inoculation with a pathogenic strain of B. bovis (BbovUFV1 7th passage). Erythrocytes of infected animals were tested in order to observer capacity of binding to BUVECs and its adhesion kinetics. The same adhesion tests were made on BUVECs stimulated with plasma of animals infected with B. bovis and culture supernatant of bovine PBMC, stimulated with the synthetic peptide RAP-1 of B. bovis containing quantified cytokines (IFN-γ, TNF-α and IL-12). There was a significant increase in the adhesion of erythrocytes of animals inoculated in BUVECs stimulated with plasma and supernatant of PBMC. However, adhesion was observed only on non-parasitised erythrocytes, suggesting that free antigens of B. bovis in the serum can prime erythrocytes non-parasitised, or still a possible expression of an isoform of VESA-1 non adherent. Adherence was not observed in the tests with samples of the negative animals. Cells stimulated with infected animals xiiiplasma and with supernatant of PBMC showed stronger expression of ICAM- 1, VCAM, PECAM-1, E-selectine and thrombospondin, them cells that didn't receive stimuli. In the same way, it was observed strong expression of ICAM- 1, VCAM, PECAM-1, E-selectine and thrombospondin in tissue samples of brain, lung and kidney in bovines infected with B. bovis, when comparing to the control group. These results suggest that Interleukins, liberated in the acute phase the Babesiosis, stimulate the expression of adhesion molecules related to the physiopathology of Babesiosis caused by B. bovis, as demonstrated by the expression of molecules in BUVECs and erythrocytes cytoadhesion. These date demonstrate physiopathologycal similarities between B. bovis and Plasmodium falciparum.

O carrapato Rhipicephalus microplus é um ectoparasito hematófago de grande importância para a pecuária por ser responsável por perdas massivas na produção animal, de forma que o seu controle é economicamente relevante. Este carrapato, além dos danos que causa por si só, é também um importante vetor para a transmissão de microorganismos patogênicos, entre eles o hemoprotozoário intraeritrocítico Babesia bovis. O presente trabalho descreve o desenvolvimento de uma linhagem de B. bovis capaz de expressar um antígeno protetor, uma glutationa S-transferase do carrapato Haemaphysalis longicornis (HlGST), e o teste desta linhagem como uma vacina de vetor vivo para o controle do carrapato R. microplus. B. bovis, em cultivo, da linhagem S74-T3B foram eletroporados em presença de plasmídeo contendo o promotor bidirecional de B. bovis Ef-1 aresponsável pela expressão independente de dois genes: o repórter fusionado ao agente para seleção (GFP-BSD) e HlGST fusionada à sequência codificadora do peptídeo sinal de MSA-1 (merozoite surface antigen-1). Após a eletroporação, foi feita a seleção com blasticidina para obtenção da linhagem nomeada HlGST. A linhagem HlGST é composta por parasitos contendo diferentes padrões de inserção dos genes exógenos, tanto dentro quanto fora do locus Ef-1. Uma linhagem clonal denominada HlGST-Cln expressando HlGST e GFP-BSD foi obtida a partir da linhagem HlGST. Dois ensaios, independentes, de imunização de bovinos com os parasitos clonais foram realizados, sendo usado como controle uma linhagem clonal previamente caracterizada denominada GFP-Cln. Todos os animais inoculados desenvolveram uma forma branda de babesiose, indicando que ambas as linhagens clonais são atenuadas, mas apenas os animais imunizados com a linhagem HlGST-Cln foram capazes de produzir anticorpos anti-HlGST. O segundo procedimento de imunização foi seguido por um desafio com larvas de R. microplus. O desenvolvimento dessas larvas no hospedeiro levou a fêmeas adultas de menor peso e fertilidade. Coletivamente, esses dados mostram a possibilidade de uso de linhagens transfectadas de B. bovis como vacinas de vetor vivo. / The tick Rhipicephalus microplus is a notorious blood-feeding ectoparasite of cattle, responsible for massive losses in animal production. It is the main vector of pathogenic microorganisms, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74- T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1 promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD, and HlGST fused to the MSA-1 (merozoite surface antigen-1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1 a locus. A B. bovis clonal line termed HlGST-Cln expressing HlGST and GFP-BSD was then derived from the mixed parasite line HlGST. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1 locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis indicating that both transfected cloned parasite lines are attenuated. All animals immunized with HlGST-Cln produced detectable anti-glutathione-S-transferase antibodies. After immunization with HlGST-Cln, calves were challenged with R. microplus larva. Development of these larva produce fully engorged female tick with reduced weight and fertility. Collectively, these data show that transfected B. bovis parasites can be used as vectors in live vectored vaccines.

Made available in DSpace on 2014-06-11T19:32:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-01-12Bitstream added on 2014-06-13T18:44:22Z : No. of bitstreams: 1 andre_mr_dr_jabo.pdf: 1327619 bytes, checksum: 69c97d185f41eb670b28bd9bfcbe3759 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Doenças transmitidas por vetores artrópodes são mundialmente importantes para a saúde humana e animal. Recentemente, diversos estudos têm sido realizados a fim de investigar o possível papel dos animais selvagens na epidemiologia destas enfermidades. A identificação de reservatórios selvagens para estes hemoparasitas ajudaria no entendimento da epi enfermidades por eles causadas, principalmente aquelas de caráter zoonótico. O presente estudo teve como objetivo pesquisar em amostras de sangue de carnívoros selvagens mantidos em cativeiro a presença de infecção ou exposição a agentes Anaplasmataceae (Ehrlichia canis, E. chaffeensis, E. ewingii, Neorickettsia risticii, N. helminthoeca, Anaplasma platys e A. phagocytophilum), Rickettsiaceae (Orientia tsutsugamushi e Rickettsia sp. dos Grupos da Febre Maculosa e do Tifo), Babesia sp., Cytauxzoon sp. (somente para os felídeos), micoplasmas hemotróficos (Mycoplasma haemofelis, Candidatus M. haemominutum, Candidatus M. turicensis, M. haemocanis e Candidatus M. haematoparvum) e Hepatozoon sp., utilizando métodos sorológicos e moleculares. Para tal, foram amostrados 167 felídeos e 100 canídeos selvagens mantidos em cativeiro nos estados de São Paulo, Mato Grosso e Distrito Federal. Doze felídeos (7,2%) e três (3%) canídeos mostraram-se soropositivos frente ao antígeno de E. canis. Apenas um canídeo (1%) mostrou-se soropositivo para A. phagocytophilum. Nenhum animal mostrou-se soropositivo para E. chaffeensis ou N. risticii. A análise filogenética baseada em um fragmento de 350 pb do gene 16S rRNA não foi suficientemente robusta para diferenciar entre as espécies de Ehrlichia spp. Os fragmentos de DNA de Ehrlichia spp. encontrados em quatro felídeos foram posicionados em um ramo distinto daquela de E. canis e E. chaffeensis, com base... / Vector-borne diseases are worldwide important to human and animal health. Recently, several studies have been done aiming to investigate the role of wild animals in the epidemiology of these diseases. The identification of wild reservoirs for these hemoparasites would help in a better understanding of the epidemiology of these diseases, mainly that with zoonotic character. The present work aimed to investigate the presence of infection or exposure to Anaplasmataceae (Ehrlichia canis, E. chaffeensis, E. ewingii, Neorickettsia risticii, N. helminthoeca, Anaplasma platys e A. phagocytophilum) and Rickettsiaceae (Orientia tsutsugamushi and Spotted Fever and Typhus Group Rickettsia sp.), Babesia sp., Cytauxzoon sp., hemotrophic hemoplasmas (Mycoplasma haemofelis, Candidatus M. haemominutum, Candidatus M. turicensis, M. haemocanis e Candidatus M. haematoparvum) and Hepatozoon sp. in captive wild carnivores blood samples by molecular and serological methods. Blood samples were collected from 167 wild felids and 100 wild canids, maintained in captivity in zoos from São Paulo and Mato Grosso states, and Distrito Federal. Twelve felids (7.2%) and three canids (3%) were seropositive to E. canis. Only one canid (1%) was seropositive to A. phagocytophilum. None was seropositive to E. chaffeensis neither N. risticii. The phylogenetic analysis of 350bp of 16S rRNA gene was not sufficiently robust to support the differentiation of Ehrlichia species. The Ehrlichia spp. DNA found in four felids was in a distinct clade from E. chaffeensis and E. canis by omp-1 phylogenetic analysis. The phylogenetic analysis of dsb gene showed the infection by E. canis in one sampled crab-eating fox. Seventeen felids (10%) and ten canids (10%) was positive to Anaplasma spp. PCR, which was in the same clade than A. phagocytophilum and A. platys by... (Complete abstract click electronic access below)