Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul

Detalhes bibliográficos
Ano de defesa: 2025
Autor(a) principal: Ries, Ananda Segabinazzi
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Santa Maria
Brasil
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
PCR
Link de acesso: http://repositorio.ufsm.br/handle/1/37210
Resumo: Protozoa can cause systemic disease and central nervous system (CNS) alterations in dogs. Among them, Toxoplasma gondii, Neospora caninum, Sarcocystis spp. (Apicomplexa), and Trypanosoma evansi are of clinical and sanitary relevance. On histopathology, Sarcocystis spp., T. gondii, and N. caninum may exhibit very similar morphology, which hampers distinction without complementary methods. In this context, this thesis combined the molecular investigation of N. caninum, T. gondii, and Sarcocystis spp. in canine brains with the characterization of a T. evansi outbreak in working dogs. In the first study, molecular detection of Apicomplexa was performed in brains from dogs submitted for necropsy at two laboratories in Rio Grande do Sul, Brazil, between 2008 and 2023, organized into two groups: G1 (n = 11), formalin-fixed, paraffin-embedded tissue with a histopathological diagnosis of protozoal encephalitis; and G2 (n = 219), frozen tissue from dogs without a history of neurological disease. DNA was extracted using commercial kits, and all samples were tested by PCR targeting the 18S rRNA gene. In G1 samples, PCR–RFLP was applied to differentiate among the investigated agents. Protozoan DNA was detected in 6/11 G1 samples - two compatible with Neospora caninum, two with Toxoplasma gondii, one with Sarcocystis neurona, and one with Sarcocystis spp. In G2, 82/219 (37.45%) samples were positive, indicating circulation of these agents also in dogs without neurological signs. These findings support the use of complementary techniques such as PCR to confirm the etiology and reduce diagnostic uncertainty when morphology is similar, and also highlight the circulation of these agents in animals without neurological signs. In the second study, an outbreak of Trypanosoma evansi was investigated in 34 dogs used in wild boar management. The animals underwent clinical examination, complete blood count, parasitological detection in blood smears, PCR and serology for Trypanosoma spp.; parasitological detection was observed in eight dogs, molecular confirmation in six, and no seropositive animals were identified. Two dogs died and were necropsied. The set of epidemiological evidence from the outbreak (direct exposure to blood and viscera during handling, with temporal clustering of cases), together with precedents of oral transmission in dogs, is consistent with infection through ingestion of blood and/or tissues from infected wild boar. Of the 34 dogs treated with diminazene aceturate, 18 improved after the first administration; the remaining 14 received a second dose, after which 10 improved and four died. This study underscores the need to include T. evansi in the diagnostic work-up of working dogs exposed during wildlife management and to adopt preventive measures to reduce transmission risk. The studies in this thesis demonstrate that: (i) PCR targeting 18S rRNA in brain tissue, together with PCR–RFLP, assists etiologic confirmation of T. gondii, N. caninum, and Sarcocystis spp.; (ii) these agents are also detected in dogs without a history of neurological signs; (iii) dogs used in wild boar management can become infected by T. evansi, with clinical and pathological impact; and (iv) incorporating these protozoa into the differential diagnosis of canine CNS diseases, and implementing surveillance and preventive actions, are necessary measures aligned with the One Health perspective.
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spelling Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do SulDetection of Toxoplasma gondii, Neospora caninum, Sarcocystis spp. and Trypanosoma spp. in canine samples in Rio Grande do SulCãesSistema nervoso centralToxoplasma gondiiNeospora caninumSarcocystis spp.Trypanosoma evansiPCRPCR-RFLPJavalisDogsCentral nervous systemWild boarCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAProtozoa can cause systemic disease and central nervous system (CNS) alterations in dogs. Among them, Toxoplasma gondii, Neospora caninum, Sarcocystis spp. (Apicomplexa), and Trypanosoma evansi are of clinical and sanitary relevance. On histopathology, Sarcocystis spp., T. gondii, and N. caninum may exhibit very similar morphology, which hampers distinction without complementary methods. In this context, this thesis combined the molecular investigation of N. caninum, T. gondii, and Sarcocystis spp. in canine brains with the characterization of a T. evansi outbreak in working dogs. In the first study, molecular detection of Apicomplexa was performed in brains from dogs submitted for necropsy at two laboratories in Rio Grande do Sul, Brazil, between 2008 and 2023, organized into two groups: G1 (n = 11), formalin-fixed, paraffin-embedded tissue with a histopathological diagnosis of protozoal encephalitis; and G2 (n = 219), frozen tissue from dogs without a history of neurological disease. DNA was extracted using commercial kits, and all samples were tested by PCR targeting the 18S rRNA gene. In G1 samples, PCR–RFLP was applied to differentiate among the investigated agents. Protozoan DNA was detected in 6/11 G1 samples - two compatible with Neospora caninum, two with Toxoplasma gondii, one with Sarcocystis neurona, and one with Sarcocystis spp. In G2, 82/219 (37.45%) samples were positive, indicating circulation of these agents also in dogs without neurological signs. These findings support the use of complementary techniques such as PCR to confirm the etiology and reduce diagnostic uncertainty when morphology is similar, and also highlight the circulation of these agents in animals without neurological signs. In the second study, an outbreak of Trypanosoma evansi was investigated in 34 dogs used in wild boar management. The animals underwent clinical examination, complete blood count, parasitological detection in blood smears, PCR and serology for Trypanosoma spp.; parasitological detection was observed in eight dogs, molecular confirmation in six, and no seropositive animals were identified. Two dogs died and were necropsied. The set of epidemiological evidence from the outbreak (direct exposure to blood and viscera during handling, with temporal clustering of cases), together with precedents of oral transmission in dogs, is consistent with infection through ingestion of blood and/or tissues from infected wild boar. Of the 34 dogs treated with diminazene aceturate, 18 improved after the first administration; the remaining 14 received a second dose, after which 10 improved and four died. This study underscores the need to include T. evansi in the diagnostic work-up of working dogs exposed during wildlife management and to adopt preventive measures to reduce transmission risk. The studies in this thesis demonstrate that: (i) PCR targeting 18S rRNA in brain tissue, together with PCR–RFLP, assists etiologic confirmation of T. gondii, N. caninum, and Sarcocystis spp.; (ii) these agents are also detected in dogs without a history of neurological signs; (iii) dogs used in wild boar management can become infected by T. evansi, with clinical and pathological impact; and (iv) incorporating these protozoa into the differential diagnosis of canine CNS diseases, and implementing surveillance and preventive actions, are necessary measures aligned with the One Health perspective.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESProtozoários podem causar doença sistêmica e alterações do sistema nervoso central (SNC) em cães. Entre eles, Toxoplasma gondii, Neospora caninum, Sarcocystis spp. (Apicomplexa) e Trypanosoma evansi são de interesse clínico e sanitário. Em histopatologia, Sarcocystis spp., T. gondii e N. caninum podem ter morfologia muito semelhante, o que dificulta a distinção sem métodos complementares de diagnóstico. Considerando esses aspectos, esta tese combinou a investigação molecular de N. caninum, T. gondii e Sarcocystis spp. em encéfalos de cães e a caracterização de um surto por T. evansi em cães de trabalho. No primeiro estudo, foi realizada a detecção molecular de Apicomplexa em encéfalos de cães submetidos à necropsia em dois laboratórios no Rio Grande do Sul entre 2008 e 2023, os quais foram organizados em dois grupos: G1 (n = 11): tecido fixado em formol, emblocado em parafina com diagnóstico histopatológico de encefalite por protozoários, e G2 (n = 219): tecido congelado de cães sem histórico de doença neurológica. O DNA foi extraído com kits comerciais e todas as amostras foram testadas por PCR com alvo no gene 18S rRNA. Nas amostras do G1, aplicou-se PCRRFLP para diferenciação entre os agentes pesquisados. Detectou-se DNA de protozoários em 6/11 amostras do G1, sendo duas compatíveis com Neospora caninum, duas com Toxoplasma gondii, uma com Sarcocystis neurona e uma com Sarcocystis spp. Do G2, 82/219 (37,45%) amostras foram positivas, indicando circulação desses agentes também em cães sem sinais neurológicos. Esses achados apoiam o uso de técnicas complementares como a PCR para confirmar a etiologia e reduzir incertezas diagnósticas quando a morfologia é semelhante, assim como alertam para circulação desses agentes em animais sem sinais neurológicos. No segundo estudo, foi investigado um surto de T. evansi em 34 cães utilizados no manejo de javalis. Os animais passaram por exame clínico, hemograma, detecção de hemoparasitas em esfregaço sanguíneo, PCR e sorologia para Trypanosoma spp., com detecção parasitológica em oito cães, confirmação molecular em seis e ausência de soropositividade. Desses, dois foram a óbito e submetidos à necropsia. O conjunto de evidências epidemiológicas do surto (exposição direta a sangue e vísceras durante o manejo, com agrupamento temporal de casos) aliado a precedentes de transmissão oral em cães é compatível com infecção por ingestão de sangue e/ou tecidos de javalis infectados. Dos 34 cães tratados com aceturato de diminazeno, 18 melhoraram após a primeira aplicação; os 14 restantes receberam uma segunda, após a qual 10 melhoraram e 4 foram a óbito. O estudo reforça a necessidade de incluir T. evansi no diagnóstico de cães de trabalho expostos ao manejo de fauna e de adotar medidas preventivas para reduzir o risco de transmissão. Os estudos desta tese contribuem ao demonstrar que: i) a PCR para 18S rRNA em encéfalos, associada ao PCR-RFLP auxilia na confirmação etiológica de T. gondii, N. caninum e Sarcocystis spp.; ii) há detecção desses agentes também em cães sem histórico de sinais neurológicos; iii) cães usados no manejo de javalis podem se infectar por T. evansi, com impacto clínico e patológico; e iv) a incorporação desses protozoários no diagnóstico diferencial de doenças do SNC e a implementação de ações de vigilância e prevenção são medidas necessárias, em alinhamento com a perspectiva de Saúde Única.Universidade Federal de Santa MariaBrasilUFSMPrograma de Pós-Graduação em Medicina VeterináriaCentro de Ciências RuraisVogel, Fernanda Silveira Flôreshttp://lattes.cnpq.br/9676833435314493Cargnelutti, Juliana FelipettoCamillo, GiovanaDalla Rosa, LucianaLudwig, Marta Helena Machado AlvesRies, Ananda Segabinazzi2026-01-06T12:01:13Z2026-01-06T12:01:13Z2025-11-18info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://repositorio.ufsm.br/handle/1/37210porAttribution-NonCommercial-NoDerivatives 4.0 Internationalinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2026-01-06T12:01:13Zoai:repositorio.ufsm.br:1/37210Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/PUBhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.com||manancial@ufsm.bropendoar:2026-01-06T12:01:13Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
Detection of Toxoplasma gondii, Neospora caninum, Sarcocystis spp. and Trypanosoma spp. in canine samples in Rio Grande do Sul
title Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
spellingShingle Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
Ries, Ananda Segabinazzi
Cães
Sistema nervoso central
Toxoplasma gondii
Neospora caninum
Sarcocystis spp.
Trypanosoma evansi
PCR
PCR-RFLP
Javalis
Dogs
Central nervous system
Wild boar
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
title_full Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
title_fullStr Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
title_full_unstemmed Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
title_sort Detecção de Toxoplasma gondii, Neospora caninum, Sarcocystis spp. e Trypanosoma spp. em amostras de cães do Rio Grande do Sul
author Ries, Ananda Segabinazzi
author_facet Ries, Ananda Segabinazzi
author_role author
dc.contributor.none.fl_str_mv Vogel, Fernanda Silveira Flôres
http://lattes.cnpq.br/9676833435314493
Cargnelutti, Juliana Felipetto
Camillo, Giovana
Dalla Rosa, Luciana
Ludwig, Marta Helena Machado Alves
dc.contributor.author.fl_str_mv Ries, Ananda Segabinazzi
dc.subject.por.fl_str_mv Cães
Sistema nervoso central
Toxoplasma gondii
Neospora caninum
Sarcocystis spp.
Trypanosoma evansi
PCR
PCR-RFLP
Javalis
Dogs
Central nervous system
Wild boar
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic Cães
Sistema nervoso central
Toxoplasma gondii
Neospora caninum
Sarcocystis spp.
Trypanosoma evansi
PCR
PCR-RFLP
Javalis
Dogs
Central nervous system
Wild boar
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Protozoa can cause systemic disease and central nervous system (CNS) alterations in dogs. Among them, Toxoplasma gondii, Neospora caninum, Sarcocystis spp. (Apicomplexa), and Trypanosoma evansi are of clinical and sanitary relevance. On histopathology, Sarcocystis spp., T. gondii, and N. caninum may exhibit very similar morphology, which hampers distinction without complementary methods. In this context, this thesis combined the molecular investigation of N. caninum, T. gondii, and Sarcocystis spp. in canine brains with the characterization of a T. evansi outbreak in working dogs. In the first study, molecular detection of Apicomplexa was performed in brains from dogs submitted for necropsy at two laboratories in Rio Grande do Sul, Brazil, between 2008 and 2023, organized into two groups: G1 (n = 11), formalin-fixed, paraffin-embedded tissue with a histopathological diagnosis of protozoal encephalitis; and G2 (n = 219), frozen tissue from dogs without a history of neurological disease. DNA was extracted using commercial kits, and all samples were tested by PCR targeting the 18S rRNA gene. In G1 samples, PCR–RFLP was applied to differentiate among the investigated agents. Protozoan DNA was detected in 6/11 G1 samples - two compatible with Neospora caninum, two with Toxoplasma gondii, one with Sarcocystis neurona, and one with Sarcocystis spp. In G2, 82/219 (37.45%) samples were positive, indicating circulation of these agents also in dogs without neurological signs. These findings support the use of complementary techniques such as PCR to confirm the etiology and reduce diagnostic uncertainty when morphology is similar, and also highlight the circulation of these agents in animals without neurological signs. In the second study, an outbreak of Trypanosoma evansi was investigated in 34 dogs used in wild boar management. The animals underwent clinical examination, complete blood count, parasitological detection in blood smears, PCR and serology for Trypanosoma spp.; parasitological detection was observed in eight dogs, molecular confirmation in six, and no seropositive animals were identified. Two dogs died and were necropsied. The set of epidemiological evidence from the outbreak (direct exposure to blood and viscera during handling, with temporal clustering of cases), together with precedents of oral transmission in dogs, is consistent with infection through ingestion of blood and/or tissues from infected wild boar. Of the 34 dogs treated with diminazene aceturate, 18 improved after the first administration; the remaining 14 received a second dose, after which 10 improved and four died. This study underscores the need to include T. evansi in the diagnostic work-up of working dogs exposed during wildlife management and to adopt preventive measures to reduce transmission risk. The studies in this thesis demonstrate that: (i) PCR targeting 18S rRNA in brain tissue, together with PCR–RFLP, assists etiologic confirmation of T. gondii, N. caninum, and Sarcocystis spp.; (ii) these agents are also detected in dogs without a history of neurological signs; (iii) dogs used in wild boar management can become infected by T. evansi, with clinical and pathological impact; and (iv) incorporating these protozoa into the differential diagnosis of canine CNS diseases, and implementing surveillance and preventive actions, are necessary measures aligned with the One Health perspective.
publishDate 2025
dc.date.none.fl_str_mv 2025-11-18
2026-01-06T12:01:13Z
2026-01-06T12:01:13Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
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rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Brasil
UFSM
Programa de Pós-Graduação em Medicina Veterinária
Centro de Ciências Rurais
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
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institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
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