Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes

Detalhes bibliográficos
Ano de defesa: 2018
Autor(a) principal: Calil, Priscila Rodrigues
Orientador(a): Ramos, Patrícia Locosque lattes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de São Carlos
Câmpus São Carlos
Programa de Pós-Graduação: Programa de Pós-Graduação em Conservação da Fauna - PPGCFau
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
PCR
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: https://repositorio.ufscar.br/handle/20.500.14289/9662
Resumo: The genus Hepatozoon represents one of six genera in hemogregarine group, sharing a similar morphology and heteroxenus life cycle (Smith,1996). Some studies in snakes indicated effects in the host, since light influence on fitness to severe effects on the growth, reproduction and offspring survival rates (Madsen et al., 2005; Brown et al., 2006). The diagnosis and identification are usually through blood smears analysis and parasite visualization; however, these results cannot be reliable in negative cases. Based on this, this study established an adapted molecular protocol to identify Hepatozoon spp., to be used as complementary test at diagnosis routine of Clinical Analyses Laboratory at São Paulo Zoological Park Foundation. The study was conducted with 117 individuals of 32 species. Microscopy and molecular techniques were used to identify the parasite. Microscopical analyses showed 13.6% of positivity. The first pair of primers, used to amplify 18S gene, show a positivity of 6.3% and the second pair of primer that amplifies the Apicoplast target, was used in the positive samples under microscopy and shows 93.33% of positivity. After sequencing, philogenetics analyses show five clusters for 18S gene and five clusters for Apicoplast target. Studies involving Hepatozoon spp. are still scarce and limited, mainly with snakes and their actual impacts on the host, so diagnostic studies are essential for the wildlife conservation, especially Ex Situ.
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spelling Calil, Priscila RodriguesRamos, Patrícia Locosquehttp://lattes.cnpq.br/7542477918045216Chagas, Carolina Romeiro Fernandeshttp://lattes.cnpq.br/5646852838444379http://lattes.cnpq.br/78642615704021780f01102c-225b-494b-8559-8bfb44451e152018-04-04T14:12:15Z2018-04-04T14:12:15Z2018-02-27CALIL, Priscila Rodrigues. Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes. 2018. Dissertação (Mestrado em Conservação da Fauna) – Universidade Federal de São Carlos, São Carlos, 2018. Disponível em: https://repositorio.ufscar.br/handle/20.500.14289/9662.https://repositorio.ufscar.br/handle/20.500.14289/9662The genus Hepatozoon represents one of six genera in hemogregarine group, sharing a similar morphology and heteroxenus life cycle (Smith,1996). Some studies in snakes indicated effects in the host, since light influence on fitness to severe effects on the growth, reproduction and offspring survival rates (Madsen et al., 2005; Brown et al., 2006). The diagnosis and identification are usually through blood smears analysis and parasite visualization; however, these results cannot be reliable in negative cases. Based on this, this study established an adapted molecular protocol to identify Hepatozoon spp., to be used as complementary test at diagnosis routine of Clinical Analyses Laboratory at São Paulo Zoological Park Foundation. The study was conducted with 117 individuals of 32 species. Microscopy and molecular techniques were used to identify the parasite. Microscopical analyses showed 13.6% of positivity. The first pair of primers, used to amplify 18S gene, show a positivity of 6.3% and the second pair of primer that amplifies the Apicoplast target, was used in the positive samples under microscopy and shows 93.33% of positivity. After sequencing, philogenetics analyses show five clusters for 18S gene and five clusters for Apicoplast target. Studies involving Hepatozoon spp. are still scarce and limited, mainly with snakes and their actual impacts on the host, so diagnostic studies are essential for the wildlife conservation, especially Ex Situ.O gênero Hepatozoon representa um dos seis gêneros do grupo das hemogregarinas, porém todos são semelhantes morfologicamente e possuem ciclo heteroxênico (Smith,1996). Alguns estudos em serpentes indicaram efeitos diversos no hospedeiro, desde leve influência no fitness até efeitos severos na taxa de crescimento, reprodução e sobrevivência dos filhotes (Madsen et al., 2005; Brown et al., 2006). O diagnóstico e identificação são realizados através da visualização do parasita em esfregaços sanguíneos, porém os resultados podem não ser confiáveis em casos negativos. Com base nisso, o presente trabalho estabeleceu um protocolo adaptado de diagnóstico molecular de Hepatozoon spp. para ser aplicado como método complementar em rotinas diagnósticas do Laboratório de Análises Clínicas na Fundação parque Zoológico de são Paulo. O estudo foi realizado com 117 indivíduos de 32 espécies. Técnicas de microscopia e biologia molecular foram utilizadas para identificação desse parasita. As análises microscópicas apresentaram positividade de 13,6%. O primeiro par de primer utilizado, que amplifica o gene 18S, obteve uma positividade de 6,3% e o segundo par de primer que amplifica o “target” Apicoplasto, foi utilizado nas amostras positivas na microscopia, obteve 93,33% de positivos. Após sequenciamento, as análises filogenéticas apresentam cinco agrupamentos para a árvore filogenética do gene 18S e cinco agrupamentos para o “target” Apicoplasto. Estudos com Hepatozoon spp. ainda são escassos e limitados, principalmente com serpentes e de seus reais impactos no hospedeiro, assim estudos de diagnóstico se mostram essenciais para a conservação das espécies, principalmente ex situ.OutraporUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Conservação da Fauna - PPGCFauUFSCarPCRCativeiroRépteisHemoparasitasHepatozoonCaptiveReptilesHemoparasitesCIENCIAS BIOLOGICASProtocolo de detecção molecular para identificação de Hepatozoon spp em serpentesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline6006006d2fac0d-222b-4530-9369-582865a28803info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARLICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstreams/56d835eb-5e08-424d-96cc-0572d7d8360a/downloadae0398b6f8b235e40ad82cba6c50031dMD55falseAnonymousREADORIGINALCALIL_Priscila_2018.pdfCALIL_Priscila_2018.pdfapplication/pdf4672136https://repositorio.ufscar.br/bitstreams/ef5fa915-b1d1-429d-9734-6bd3a869c7a8/downloadadbca81aab9a0b0c3c628e32e54f7aa7MD56trueAnonymousREADTEXTCALIL_Priscila_2018.pdf.txtCALIL_Priscila_2018.pdf.txtExtracted texttext/plain203786https://repositorio.ufscar.br/bitstreams/3e83c0b0-9970-4028-a581-1a0ea69af557/download5c7e234621b2ba6d1d632870fae9af00MD59falseAnonymousREADTHUMBNAILCALIL_Priscila_2018.pdf.jpgCALIL_Priscila_2018.pdf.jpgIM Thumbnailimage/jpeg3455https://repositorio.ufscar.br/bitstreams/1eb57d99-1d19-4ac9-bc54-4fea5a2d932c/download5add0265455b083e54bcedf28f1eb622MD510falseAnonymousREAD20.500.14289/96622025-02-05 17:49:19.157Acesso abertoopen.accessoai:repositorio.ufscar.br:20.500.14289/9662https://repositorio.ufscar.brRepositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestrepositorio.sibi@ufscar.bropendoar:43222025-02-05T20:49:19Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)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
dc.title.por.fl_str_mv Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
title Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
spellingShingle Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
Calil, Priscila Rodrigues
PCR
Cativeiro
Répteis
Hemoparasitas
Hepatozoon
Captive
Reptiles
Hemoparasites
CIENCIAS BIOLOGICAS
title_short Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
title_full Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
title_fullStr Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
title_full_unstemmed Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
title_sort Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes
author Calil, Priscila Rodrigues
author_facet Calil, Priscila Rodrigues
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/7864261570402178
dc.contributor.author.fl_str_mv Calil, Priscila Rodrigues
dc.contributor.advisor1.fl_str_mv Ramos, Patrícia Locosque
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/7542477918045216
dc.contributor.advisor-co1.fl_str_mv Chagas, Carolina Romeiro Fernandes
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/5646852838444379
dc.contributor.authorID.fl_str_mv 0f01102c-225b-494b-8559-8bfb44451e15
contributor_str_mv Ramos, Patrícia Locosque
Chagas, Carolina Romeiro Fernandes
dc.subject.por.fl_str_mv PCR
Cativeiro
Répteis
Hemoparasitas
topic PCR
Cativeiro
Répteis
Hemoparasitas
Hepatozoon
Captive
Reptiles
Hemoparasites
CIENCIAS BIOLOGICAS
dc.subject.eng.fl_str_mv Hepatozoon
Captive
Reptiles
Hemoparasites
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS
description The genus Hepatozoon represents one of six genera in hemogregarine group, sharing a similar morphology and heteroxenus life cycle (Smith,1996). Some studies in snakes indicated effects in the host, since light influence on fitness to severe effects on the growth, reproduction and offspring survival rates (Madsen et al., 2005; Brown et al., 2006). The diagnosis and identification are usually through blood smears analysis and parasite visualization; however, these results cannot be reliable in negative cases. Based on this, this study established an adapted molecular protocol to identify Hepatozoon spp., to be used as complementary test at diagnosis routine of Clinical Analyses Laboratory at São Paulo Zoological Park Foundation. The study was conducted with 117 individuals of 32 species. Microscopy and molecular techniques were used to identify the parasite. Microscopical analyses showed 13.6% of positivity. The first pair of primers, used to amplify 18S gene, show a positivity of 6.3% and the second pair of primer that amplifies the Apicoplast target, was used in the positive samples under microscopy and shows 93.33% of positivity. After sequencing, philogenetics analyses show five clusters for 18S gene and five clusters for Apicoplast target. Studies involving Hepatozoon spp. are still scarce and limited, mainly with snakes and their actual impacts on the host, so diagnostic studies are essential for the wildlife conservation, especially Ex Situ.
publishDate 2018
dc.date.accessioned.fl_str_mv 2018-04-04T14:12:15Z
dc.date.available.fl_str_mv 2018-04-04T14:12:15Z
dc.date.issued.fl_str_mv 2018-02-27
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv CALIL, Priscila Rodrigues. Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes. 2018. Dissertação (Mestrado em Conservação da Fauna) – Universidade Federal de São Carlos, São Carlos, 2018. Disponível em: https://repositorio.ufscar.br/handle/20.500.14289/9662.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/20.500.14289/9662
identifier_str_mv CALIL, Priscila Rodrigues. Protocolo de detecção molecular para identificação de Hepatozoon spp em serpentes. 2018. Dissertação (Mestrado em Conservação da Fauna) – Universidade Federal de São Carlos, São Carlos, 2018. Disponível em: https://repositorio.ufscar.br/handle/20.500.14289/9662.
url https://repositorio.ufscar.br/handle/20.500.14289/9662
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dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
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publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
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