Estudo da associação entre Angiostrongylus vasorum com Achatina fulica

Detalhes bibliográficos
Autor(a) principal: Aytube Coaglio Silva Lucas
Data de Publicação: 2017
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFMG
Texto Completo: http://hdl.handle.net/1843/34822
Resumo: Angiostrongylus vasorum is a parasitic nematode of domestic and wild canids, whose adult form can be found in the right ventricle, pulmonary arteries and their ramifications. The cycle involves is heteroxene with the participation of aquatic and terrestrial molluscs as intermediate hosts. The present study evaluated the relationship between Achatina fulica experimentally infected with 1000 L1 of A. vasorum on days 0, 1º, 8º, 13º and 30º after infection (dpi). The presence of phenoloxidase (PO) in the hemolymph acellular fraction in the hemocyte supernatant was analyzed using the L-DOPA, L-Tyrosine and p-phenylenediamine (PPD) substrates, as well as the evidence of melanin in the tissue of the region cephalopodal. The level of nitric oxide in the acellular fraction of A. fulica was quantified by the Griess reaction. In this study, the morphology and morphometry of L1, L2 and L3 were evaluated and fixed with 4% paraformaldehyde and labeled with antibodies: anti-β-tubulin, anti-α-tubulin, anti-collagen 1/1-collagen and anti-β-actin, and by the dyes: picrosirius and oil red, under an epifluorescence microscope. To observe the natural infection by A. vasorum was carried out the chemical digestion of A. fulica captured in the city of Belo Horizonte, Sabará, Igarapé and Lagoa Santa. PO activity and tissue melanin production were observed in A. fulica, varying between control and infected. At the 1st dpi, L-tyrosine (p <0.001), L-DOPA (p <0.05) and PPD (p <0.01) were observed, increased PO coincident with melanin. At 8° dpi, modulation of L-DOPA-dependent PO (p <0.001) and L-Tyrosine, but not with PPD (p <0.05) was observed, while in the tissue analyzes they were inversely proportional to PO on substrates L-DOPA and L-Tyrosine. At 13° dpi the inhibition of PO occurred in all three substrates and circulating levels of melanin in the tissue decreased. At 30° dpi the PO with L-Tyrosine did not change, whereas for L-DOPA and PPD they increased and in the tissue there was a decrease in the concentration of melanin. At the level of NO, it increased up to 8° dpi and decreased to 30° dpi. The larvae L1, L2, L3 of A. vasorum present in the cytokinetic composition fibers of actin, tubulin and collagen. With picrosirus, in the epifluorescence associated with ApoTome obervou internal structures in the anterior and posterior region. In the oil red, lipids were found to be dispersed throughout L1, L2 the lipid reserves fill the entire length of the larva and L3 were located in the posterior part of the larvae. In the molluscs of the metropolitan region of Belo Horizonte, only larvae of the genus Strongyluris sp and Rhaditis sp. were found. The work presents a possible mechanism, phenoloxidase, in the elimination of larvae of A. vasorum, mainly in the L1 to L2 stage, where there is an increase of nitric oxide and melanin in the tissues. Picrossirius was the used dye that presented the best result for morphological analysis of the larvae, having the advantage of being cheap and easy to access. The molluscs collected in Belo Horizonte and in the metropolitan region were not infected with metatrongilid larvae, and could indicate the absence of the parasites in these places, being areas free for metastrongilideos.
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spelling Estudo da associação entre Angiostrongylus vasorum com Achatina fulicaParasitologiaAngiostrongylus vasorumAchatina fulicaParasitologiaDirofilarioseMoluscosAngiostrongylus vasorum is a parasitic nematode of domestic and wild canids, whose adult form can be found in the right ventricle, pulmonary arteries and their ramifications. The cycle involves is heteroxene with the participation of aquatic and terrestrial molluscs as intermediate hosts. The present study evaluated the relationship between Achatina fulica experimentally infected with 1000 L1 of A. vasorum on days 0, 1º, 8º, 13º and 30º after infection (dpi). The presence of phenoloxidase (PO) in the hemolymph acellular fraction in the hemocyte supernatant was analyzed using the L-DOPA, L-Tyrosine and p-phenylenediamine (PPD) substrates, as well as the evidence of melanin in the tissue of the region cephalopodal. The level of nitric oxide in the acellular fraction of A. fulica was quantified by the Griess reaction. In this study, the morphology and morphometry of L1, L2 and L3 were evaluated and fixed with 4% paraformaldehyde and labeled with antibodies: anti-β-tubulin, anti-α-tubulin, anti-collagen 1/1-collagen and anti-β-actin, and by the dyes: picrosirius and oil red, under an epifluorescence microscope. To observe the natural infection by A. vasorum was carried out the chemical digestion of A. fulica captured in the city of Belo Horizonte, Sabará, Igarapé and Lagoa Santa. PO activity and tissue melanin production were observed in A. fulica, varying between control and infected. At the 1st dpi, L-tyrosine (p <0.001), L-DOPA (p <0.05) and PPD (p <0.01) were observed, increased PO coincident with melanin. At 8° dpi, modulation of L-DOPA-dependent PO (p <0.001) and L-Tyrosine, but not with PPD (p <0.05) was observed, while in the tissue analyzes they were inversely proportional to PO on substrates L-DOPA and L-Tyrosine. At 13° dpi the inhibition of PO occurred in all three substrates and circulating levels of melanin in the tissue decreased. At 30° dpi the PO with L-Tyrosine did not change, whereas for L-DOPA and PPD they increased and in the tissue there was a decrease in the concentration of melanin. At the level of NO, it increased up to 8° dpi and decreased to 30° dpi. The larvae L1, L2, L3 of A. vasorum present in the cytokinetic composition fibers of actin, tubulin and collagen. With picrosirus, in the epifluorescence associated with ApoTome obervou internal structures in the anterior and posterior region. In the oil red, lipids were found to be dispersed throughout L1, L2 the lipid reserves fill the entire length of the larva and L3 were located in the posterior part of the larvae. In the molluscs of the metropolitan region of Belo Horizonte, only larvae of the genus Strongyluris sp and Rhaditis sp. were found. The work presents a possible mechanism, phenoloxidase, in the elimination of larvae of A. vasorum, mainly in the L1 to L2 stage, where there is an increase of nitric oxide and melanin in the tissues. Picrossirius was the used dye that presented the best result for morphological analysis of the larvae, having the advantage of being cheap and easy to access. The molluscs collected in Belo Horizonte and in the metropolitan region were not infected with metatrongilid larvae, and could indicate the absence of the parasites in these places, being areas free for metastrongilideos.Angiostrongylus vasorum é um nematóide parasito de canídeos domésticos e silvestres, cuja forma adulta pode ser encontrada no ventrículo direito, artérias pulmonares e ramificações. O ciclo envolve uma fase no ambiente e a participação de moluscos aquáticos e terrestres como hospedeiros intermediários. O presente trabalho avaliou a relação entre Achatina fulica infectado experimentalmente com 1000 L1 A. vasorum, nos dias 01, 08,13 e 30. Analisou-se a presença de fenoloxidase (PO), na fração acelular da hemolinfa, no sobrenadante de hemócitos, utilizando os substratos L-Dopa L- Tirosina e p-fenilenediamine (PPD), assim como a evidência de melanina no tecido da região cefalopodal. Também quantificou-se óxido nítrico, na fração acelular de A. fulica pela Reação de Griess. Ainda, nesse estudo, avaliou-se a morfologia e morfometria de L1, L2 e L3, fixadas com paraformaldeído 4% e marcadas com anticorpos anti-β-tubulina, anti- α- tubulina, anti- colga 1/1- colagen, anti-β- actina, pelo picrosirius, oil red em microscópio de epifluorescência. E, para observar a infecção natural por A. vasorum, realizou-se digestão química de A. fulica, capturados na cidade de Belo Horizonte, Sabará, Igarapé e Lagoa Santa. Observou-se atividade de PO e produção de melanina tecidual em A. fulica, variando entre o controle e infectados. Ao 1°dpi, observou-se com L-tirosina (p<0,001), L-Dopa (p<0,05) e PPD (p<0,01) aumento de PO coincidente com melanina. Ao 8° dpi, observa-se modulação de PO dependente de L-DOPA (p<0,001) e L-tirosina, mas não com o PPD (p<0,05). Já as análises teciduais foram inversamente proporcionais à PO dosificada com L-DOPA, L-tirosina. Ao 13° dpi, a inibição de PO ocorreu para os três substratos, sendo para L-DOPA (p<0,01) e PPD (p<0,001), e os níveis circulantes de melanina diminuíram no tecido. Ao 30° dpi, a PO com L-tirosina não alterou, enquanto para L-DOPA e PPD aumentaram, e no tecido decresceu a concentração de melanina. O NO foi crescente até o 8° dpi, decrescendo até o 30° dpi. As larvas L1, L2, L3 de A. vasorum apresentam, na composição do citoesqueleto, fibras de actina, tubulina, colágeno. Com picrosirus na luz polarizada não evidenciou a morfologia das larvas. No entanto, na epifluorescência associada ao APOTOME, observaram-se estruturas internas da região anterior e posterior. Pelo Oil red, verificou-se que existem reservas de lipídeos localizadas na região anterior e posterior de L1; na L2, as reservas lipídicas preenchem toda a extensão da larva e, na L3, localiza-se na parte posterior. Nos moluscos da região metropolitana de Belo Horizonte encontrou-se, apenas, larvas do gênero Strongyluris sp e Rhaditis sp.. O trabalho apresenta um possível mecanismo, a fenoloxidase, na eliminação das larvas de A. vasorum, principalmente no estádio L1 para L2, onde há aumento de óxido nítrico e melanina nos tecidos. O picrossirius foi o corante usado que apresentou o melhor resultado para análise morfológica das larvas, tendo como vantagem de ser barato e de fácil acesso. Os moluscos coletados em Belo Horizonte e na região metropolitana não estavam infectados com larvas de metastrongilideos, podendo indicar a ausência dos parasitos nesses locais, sendo áreas livres para metastrongilideos.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoUniversidade Federal de Minas GeraisBrasilICB - INSTITUTO DE CIÊNCIAS BIOLOGICASPrograma de Pós-Graduação em ParasitologiaUFMGWalter dos Santos Limahttp://lattes.cnpq.br/8855532073043045Cíntia Aparecida de Jesus PereiraMarcos Pezzi GuimarãesAndréa Teixeira CarvalhoJoziana Muniz de Paiva BarçanteLuciano dos Santos Aggum CapettiniAytube Coaglio Silva Lucas2021-01-21T12:30:13Z2021-01-21T12:30:13Z2017-06-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/1843/34822porhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2021-01-21T12:30:13Zoai:repositorio.ufmg.br:1843/34822Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2021-01-21T12:30:13Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
title Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
spellingShingle Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
Aytube Coaglio Silva Lucas
Parasitologia
Angiostrongylus vasorum
Achatina fulica
Parasitologia
Dirofilariose
Moluscos
title_short Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
title_full Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
title_fullStr Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
title_full_unstemmed Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
title_sort Estudo da associação entre Angiostrongylus vasorum com Achatina fulica
author Aytube Coaglio Silva Lucas
author_facet Aytube Coaglio Silva Lucas
author_role author
dc.contributor.none.fl_str_mv Walter dos Santos Lima
http://lattes.cnpq.br/8855532073043045
Cíntia Aparecida de Jesus Pereira
Marcos Pezzi Guimarães
Andréa Teixeira Carvalho
Joziana Muniz de Paiva Barçante
Luciano dos Santos Aggum Capettini
dc.contributor.author.fl_str_mv Aytube Coaglio Silva Lucas
dc.subject.por.fl_str_mv Parasitologia
Angiostrongylus vasorum
Achatina fulica
Parasitologia
Dirofilariose
Moluscos
topic Parasitologia
Angiostrongylus vasorum
Achatina fulica
Parasitologia
Dirofilariose
Moluscos
description Angiostrongylus vasorum is a parasitic nematode of domestic and wild canids, whose adult form can be found in the right ventricle, pulmonary arteries and their ramifications. The cycle involves is heteroxene with the participation of aquatic and terrestrial molluscs as intermediate hosts. The present study evaluated the relationship between Achatina fulica experimentally infected with 1000 L1 of A. vasorum on days 0, 1º, 8º, 13º and 30º after infection (dpi). The presence of phenoloxidase (PO) in the hemolymph acellular fraction in the hemocyte supernatant was analyzed using the L-DOPA, L-Tyrosine and p-phenylenediamine (PPD) substrates, as well as the evidence of melanin in the tissue of the region cephalopodal. The level of nitric oxide in the acellular fraction of A. fulica was quantified by the Griess reaction. In this study, the morphology and morphometry of L1, L2 and L3 were evaluated and fixed with 4% paraformaldehyde and labeled with antibodies: anti-β-tubulin, anti-α-tubulin, anti-collagen 1/1-collagen and anti-β-actin, and by the dyes: picrosirius and oil red, under an epifluorescence microscope. To observe the natural infection by A. vasorum was carried out the chemical digestion of A. fulica captured in the city of Belo Horizonte, Sabará, Igarapé and Lagoa Santa. PO activity and tissue melanin production were observed in A. fulica, varying between control and infected. At the 1st dpi, L-tyrosine (p <0.001), L-DOPA (p <0.05) and PPD (p <0.01) were observed, increased PO coincident with melanin. At 8° dpi, modulation of L-DOPA-dependent PO (p <0.001) and L-Tyrosine, but not with PPD (p <0.05) was observed, while in the tissue analyzes they were inversely proportional to PO on substrates L-DOPA and L-Tyrosine. At 13° dpi the inhibition of PO occurred in all three substrates and circulating levels of melanin in the tissue decreased. At 30° dpi the PO with L-Tyrosine did not change, whereas for L-DOPA and PPD they increased and in the tissue there was a decrease in the concentration of melanin. At the level of NO, it increased up to 8° dpi and decreased to 30° dpi. The larvae L1, L2, L3 of A. vasorum present in the cytokinetic composition fibers of actin, tubulin and collagen. With picrosirus, in the epifluorescence associated with ApoTome obervou internal structures in the anterior and posterior region. In the oil red, lipids were found to be dispersed throughout L1, L2 the lipid reserves fill the entire length of the larva and L3 were located in the posterior part of the larvae. In the molluscs of the metropolitan region of Belo Horizonte, only larvae of the genus Strongyluris sp and Rhaditis sp. were found. The work presents a possible mechanism, phenoloxidase, in the elimination of larvae of A. vasorum, mainly in the L1 to L2 stage, where there is an increase of nitric oxide and melanin in the tissues. Picrossirius was the used dye that presented the best result for morphological analysis of the larvae, having the advantage of being cheap and easy to access. The molluscs collected in Belo Horizonte and in the metropolitan region were not infected with metatrongilid larvae, and could indicate the absence of the parasites in these places, being areas free for metastrongilideos.
publishDate 2017
dc.date.none.fl_str_mv 2017-06-28
2021-01-21T12:30:13Z
2021-01-21T12:30:13Z
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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dc.identifier.uri.fl_str_mv http://hdl.handle.net/1843/34822
url http://hdl.handle.net/1843/34822
dc.language.iso.fl_str_mv por
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dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/pt/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/pt/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
Brasil
ICB - INSTITUTO DE CIÊNCIAS BIOLOGICAS
Programa de Pós-Graduação em Parasitologia
UFMG
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
Brasil
ICB - INSTITUTO DE CIÊNCIAS BIOLOGICAS
Programa de Pós-Graduação em Parasitologia
UFMG
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
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institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv repositorio@ufmg.br
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