Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum

Detalhes bibliográficos
Ano de defesa: 2014
Autor(a) principal: Candido, Renata Russo Frasca
Orientador(a): Graeff-Teixeira, Carlos
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Pontifícia Universidade Católica do Rio Grande do Sul
Programa de Pós-Graduação: Programa de Pós-Graduação em Biologia Celular e Molecular
Departamento: Faculdade de Biociências
País: Brasil
Palavras-chave em Português:
Área do conhecimento CNPq:
Link de acesso: http://tede2.pucrs.br/tede2/handle/tede/5912
Resumo: Schistososmiasis is a chronic endemic infection caused by parasites of the genus Schistosoma, and it occurs in 74 countries in Africa, South America and Asia. The three main agents of this infection in humans are: Schistosoma mansoni and Schistosoma japonicum, that cause the hepatic-intestinal disease, and Schistosoma haematobium, responsible for the genitourinary infection. Despite the effective treatment like praziquantel, schistososmiasis remains as the second most prevalent parasitic disease in the world. Diagnosis of the intestinal schistososmiasis is achieved through the direct visualization of the eggs in fecal samples. The current method recommended by the World Health Organization in epidemiological studies is the Kato-Katz method. Despite it being simple and cheap, in areas of low endemicity this technique loose sensibility, leading to the occurrence of false-negative cases and underestimation of the prevalence in the studied area. Helmintex™ is a coproparasitological method highly sensitive that allows the isolation of Schistosoma eggs from 30 grams of feces, based in the interaction between the eggs and paramagnetic microspheres in a magnetic field. However, this method demands time and specialized equipment, being of difficult manipulation in work field. The mechanism that promotes the interaction between the paramagnetic spheres with the Schistosoma eggs is not known. Considering the necessity of sensitive diagnostic tools of easy applicability in epidemiological studies in low endemicity areas, this work has the purpose to study the surface physical-chemical characteristics of S. mansoni and S. japonicum eggs, in order to enhance the efficiency of the Helmintex™ method. S. mansoni and S. japonicum eggs were isolated from livers of experimentally infected mice. The eggs were submitted to morphological and structural analysis using Scanning and Transmission Electron Microscopy and elemental analysis using Energy Disperssion Spectroscopy. The magnetic susceptibility was determined using SQUID (Superconducting Quantum Interference Device) and the concentration of the chemical elements was determined through Atomic Emission Spectroscopy. Experiments to elucidate the interaction properties of the eggs of the eggs and the microspheres were conducted incubating the eggs from both species with different paramagnetic microspheres. The results show that the egg surface of both species is recovered by a dense layer of microspines, being those shorter and less spaced in S. mansoni. The eggs spontaneously bind the particles, with a greater preference for magnetic material. S. japonicum eggs have a higher affinity for paramagnetic microspheres than S. mansoni eggs. The presence of streptavidin in the surface of the microspheres enhances the affinity of both species for non-magnetic material, however it decreases the affinity for paramagnetic microspheres. Despite the presence of iron in the eggshell of S. mansoni and S. japonicum, the origin of the interaction does not seem to be magnetic, but, based in the difference of electrostatic charges present in the surface of the eggs and the microspheres. The continuity of this study is important to determine the physical-chemical characteristics of eggs from human feces, and it can lead to the upgrading and optimization of the Helmintex™ method. Studies using Atomic Force Microscopy are in progress.
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spelling Graeff-Teixeira, Carlos992.077.610-68Candido, Renata Russo Frasca2015-04-17T14:54:06Z2014-06-27http://tede2.pucrs.br/tede2/handle/tede/5912Schistososmiasis is a chronic endemic infection caused by parasites of the genus Schistosoma, and it occurs in 74 countries in Africa, South America and Asia. The three main agents of this infection in humans are: Schistosoma mansoni and Schistosoma japonicum, that cause the hepatic-intestinal disease, and Schistosoma haematobium, responsible for the genitourinary infection. Despite the effective treatment like praziquantel, schistososmiasis remains as the second most prevalent parasitic disease in the world. Diagnosis of the intestinal schistososmiasis is achieved through the direct visualization of the eggs in fecal samples. The current method recommended by the World Health Organization in epidemiological studies is the Kato-Katz method. Despite it being simple and cheap, in areas of low endemicity this technique loose sensibility, leading to the occurrence of false-negative cases and underestimation of the prevalence in the studied area. Helmintex™ is a coproparasitological method highly sensitive that allows the isolation of Schistosoma eggs from 30 grams of feces, based in the interaction between the eggs and paramagnetic microspheres in a magnetic field. However, this method demands time and specialized equipment, being of difficult manipulation in work field. The mechanism that promotes the interaction between the paramagnetic spheres with the Schistosoma eggs is not known. Considering the necessity of sensitive diagnostic tools of easy applicability in epidemiological studies in low endemicity areas, this work has the purpose to study the surface physical-chemical characteristics of S. mansoni and S. japonicum eggs, in order to enhance the efficiency of the Helmintex™ method. S. mansoni and S. japonicum eggs were isolated from livers of experimentally infected mice. The eggs were submitted to morphological and structural analysis using Scanning and Transmission Electron Microscopy and elemental analysis using Energy Disperssion Spectroscopy. The magnetic susceptibility was determined using SQUID (Superconducting Quantum Interference Device) and the concentration of the chemical elements was determined through Atomic Emission Spectroscopy. Experiments to elucidate the interaction properties of the eggs of the eggs and the microspheres were conducted incubating the eggs from both species with different paramagnetic microspheres. The results show that the egg surface of both species is recovered by a dense layer of microspines, being those shorter and less spaced in S. mansoni. The eggs spontaneously bind the particles, with a greater preference for magnetic material. S. japonicum eggs have a higher affinity for paramagnetic microspheres than S. mansoni eggs. The presence of streptavidin in the surface of the microspheres enhances the affinity of both species for non-magnetic material, however it decreases the affinity for paramagnetic microspheres. Despite the presence of iron in the eggshell of S. mansoni and S. japonicum, the origin of the interaction does not seem to be magnetic, but, based in the difference of electrostatic charges present in the surface of the eggs and the microspheres. The continuity of this study is important to determine the physical-chemical characteristics of eggs from human feces, and it can lead to the upgrading and optimization of the Helmintex™ method. Studies using Atomic Force Microscopy are in progress.A esquistossomose é uma infecção crônica endêmica causada por parasitos do gênero Schistosoma, e ocorre em países 74 países na África, América do Sul e Ásia. Os três principais agentes desta infecção em humanos são: Schistosoma mansoni e Schistosoma japonicum, causadores da doença hepato-intestinal, e Schistosoma haematobium, responsável pela infecção genitourinária. Apesar de haver tratamento efetivo como o praziquantel, a esquistossomose permanece como a segunda infecção parasitária mais prevalente no mundo. O diagnóstico da esquistossomose intestinal é feito através da direta visualização dos ovos em amostras fecais. O método atualmente recomendado pela Organização Mundial de Saúde em estudos epidemiológicos é o método de Kato-Katz. Apesar de simples e barato, em áreas de baixa endemicidade esta técnica perde sensibilidade, levando à ocorrência de casos falso-negativos e subestimação da prevalência da área estudada. O Helmintex® é um método coproparasitológico altamente sensível que permite o isolamento de ovos de Schistosoma à partir de 30 gramas de fezes, baseado na interação entre os ovos e esferas paramagnéticas em um campo magnético. Entretanto, este método demanda tempo e equipamentos especializados, sendo de difícil manipulação em estudos de campo. O mecanismo que promove a interação das esferas paramagnéticas com os ovos de Schistosoma não é conhecido. Tendo em vista a necessidade de ferramentas diagnósticas sensíveis e de fácil aplicabilidade em estudos epidemiológicos em áreas de baixa transmissão, este trabalho tem por objetivo estudar características físico-químicas da superfície dos ovos de S. mansoni e S. japonicum, afim de aprimorar a eficiência do método Helmintex®. Ovos de S. mansoni e S. japonicum foram isolados de fígados de camundongos experimentalmente infectados. Os ovos foram submetidos à analise morfológica e estrutural utilizando Microscopia Eletrônica de Varredura e Transmissão e análise elementar utilizando Espectroscopia por Dispersão de Energia. A susceptibilidade magnética foi determinada utilizando-se o SQUID (Superconducting Quantum Interference Device) e a concentração dos elementos químicos foi determinada através de Espectroscopia por Emissão Atômica. Experimentos para elucidar as propriedades de interação dos ovos e das microesferas foram conduzidos incubando ovos de ambas as espécies com diferentes microesferas paramagnéticas. Os resultados mostram que a superfície do ovo de ambas as espécies é recoberta por uma camada densa de microespinhos, sendo estes mais curtos e menos espaçados em S. mansoni. Os ovos espontaneamente ligam-se às partículas, com maior preferência por material magnético. Os ovos de S. japonicum possuem maior afinidade pelas microsesferas paramagnéticas do que os ovos de S. mansoni. A presença de estreptavidina na superfície das microesferas aumenta a afinidade de ambas as espécies por microesferas não-magnéticas, porém diminui a afinidade por microesferas paramagnéticas. Apesar da presença de ferro na casca do ovo tanto de S. mansoni quanto de S. japonicum, a origem da interação não parece ser magnética, e sim, baseada na diferença de cargas eletrostáticas presentes na superfície dos ovos e das microesferas. A continuidade deste estudo é importante para determinar as características físico-químicas de ovos provenientes de fezes humanas, e pode levar ao aprimoramento e otimização do método Helmintex®. Estudos utilizando-se Microscopia de Força Atômica encontram-se em andamento.Submitted by Setor de Tratamento da Informação - BC/PUCRS (tede2@pucrs.br) on 2015-04-17T14:54:06Z No. of bitstreams: 1 467370 - Texto Completo.pdf: 6072599 bytes, checksum: 169f7ff123882c0b088732b04ad1956d (MD5)Made available in DSpace on 2015-04-17T14:54:06Z (GMT). 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dc.title.por.fl_str_mv Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
title Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
spellingShingle Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
Candido, Renata Russo Frasca
BIOLOGIA MOLECULAR
BIOLOGIA CELULAR
ESQUISTOSSOMOSE
PARASITOLOGIA MÉDICA
CIENCIAS BIOLOGICAS
title_short Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
title_full Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
title_fullStr Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
title_full_unstemmed Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
title_sort Estudo das características físico-químicas e propriedades magnéticas da superfície do ovo de Schistosoma mansoni e Schistosoma japonicum
author Candido, Renata Russo Frasca
author_facet Candido, Renata Russo Frasca
author_role author
dc.contributor.advisor1.fl_str_mv Graeff-Teixeira, Carlos
dc.contributor.authorID.fl_str_mv 992.077.610-68
dc.contributor.author.fl_str_mv Candido, Renata Russo Frasca
contributor_str_mv Graeff-Teixeira, Carlos
dc.subject.por.fl_str_mv BIOLOGIA MOLECULAR
BIOLOGIA CELULAR
ESQUISTOSSOMOSE
PARASITOLOGIA MÉDICA
topic BIOLOGIA MOLECULAR
BIOLOGIA CELULAR
ESQUISTOSSOMOSE
PARASITOLOGIA MÉDICA
CIENCIAS BIOLOGICAS
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS
description Schistososmiasis is a chronic endemic infection caused by parasites of the genus Schistosoma, and it occurs in 74 countries in Africa, South America and Asia. The three main agents of this infection in humans are: Schistosoma mansoni and Schistosoma japonicum, that cause the hepatic-intestinal disease, and Schistosoma haematobium, responsible for the genitourinary infection. Despite the effective treatment like praziquantel, schistososmiasis remains as the second most prevalent parasitic disease in the world. Diagnosis of the intestinal schistososmiasis is achieved through the direct visualization of the eggs in fecal samples. The current method recommended by the World Health Organization in epidemiological studies is the Kato-Katz method. Despite it being simple and cheap, in areas of low endemicity this technique loose sensibility, leading to the occurrence of false-negative cases and underestimation of the prevalence in the studied area. Helmintex™ is a coproparasitological method highly sensitive that allows the isolation of Schistosoma eggs from 30 grams of feces, based in the interaction between the eggs and paramagnetic microspheres in a magnetic field. However, this method demands time and specialized equipment, being of difficult manipulation in work field. The mechanism that promotes the interaction between the paramagnetic spheres with the Schistosoma eggs is not known. Considering the necessity of sensitive diagnostic tools of easy applicability in epidemiological studies in low endemicity areas, this work has the purpose to study the surface physical-chemical characteristics of S. mansoni and S. japonicum eggs, in order to enhance the efficiency of the Helmintex™ method. S. mansoni and S. japonicum eggs were isolated from livers of experimentally infected mice. The eggs were submitted to morphological and structural analysis using Scanning and Transmission Electron Microscopy and elemental analysis using Energy Disperssion Spectroscopy. The magnetic susceptibility was determined using SQUID (Superconducting Quantum Interference Device) and the concentration of the chemical elements was determined through Atomic Emission Spectroscopy. Experiments to elucidate the interaction properties of the eggs of the eggs and the microspheres were conducted incubating the eggs from both species with different paramagnetic microspheres. The results show that the egg surface of both species is recovered by a dense layer of microspines, being those shorter and less spaced in S. mansoni. The eggs spontaneously bind the particles, with a greater preference for magnetic material. S. japonicum eggs have a higher affinity for paramagnetic microspheres than S. mansoni eggs. The presence of streptavidin in the surface of the microspheres enhances the affinity of both species for non-magnetic material, however it decreases the affinity for paramagnetic microspheres. Despite the presence of iron in the eggshell of S. mansoni and S. japonicum, the origin of the interaction does not seem to be magnetic, but, based in the difference of electrostatic charges present in the surface of the eggs and the microspheres. The continuity of this study is important to determine the physical-chemical characteristics of eggs from human feces, and it can lead to the upgrading and optimization of the Helmintex™ method. Studies using Atomic Force Microscopy are in progress.
publishDate 2014
dc.date.issued.fl_str_mv 2014-06-27
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