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Previous issue date: 2013-02-28 / Two species of parasites nematode of Angiostrongylidae family, genus Angiostrongylus intra-arterial localization are capable of human desease production: Angiostrongylus costaricensis and Angiostrongylus cantonensis. Both species are rodent s parasites and the human infection is considered incidental. On humans, A. cantonensis cause eosinophilic meningitis and A. costaricensis cause abdominal angiostrongiliase. Since there is no elimination of parasitic forms in human infection, diagnosis becomes difficult and molecular methods are necessary. After years of discovery of angiostrongil?ases, there are still many efforts in the study and development of a specific and sensitive diagnostic test capable of discriminating Angiostrongyliases of the other parasites. In this context, cross-reactivity becomes a problem in specificity of serologic tests. The main objective of this study was to analyze the sharing of antigens between Angiostrongylus spp. and parasites Strongyloides spp., Fasciola hepatica, Ascaris lumbricoides, Hymenolepis diminuta and Toxocara canis, and identify these molecules shared. To obtain antigens, some rats were captures and their feces analyzed by the method of Baerman addition to being seeded Agar Plates. Were obtained larvae of Strongyloides spp. and Angiostrongylus spp. In pulmonary arteries were found 11 female worms and 2 male worms of A. cantonensis, which led to the first report of the occurrence of this parasite in Rio Grande do Sul. In the small intestine was obtained a Strongyloides spp worm and analysis of small intestine were obtained Hymenolepis diminuta worms. From the adult worms of parasites were obtained antigens used in this study. Furthermore, we used antigens of allergens such as peanut, tomato, strawberry and pollen. To identify shared antigens, proteins were separated by one and two dimensional electrophoresis technique assayed by Western blot using sera of patients with angiostrongyliases. All antigens were recognized by sera from patients infected with angiostrongyliases, being a total of fourteen bands identified as being immunogenic. The fourteen bands were cut out of polyacrylamide gels and analyzed by mass spectrometry. The proteins identified were: Phosphoenolpyruvate carboxykinase, heat shock protein 70 (HSP70), DAPPUDRAFT hypothetical protein, 60 kDa Chaperonin 5, Glyceraldehyde-3-phosphate dehydrogenase, sigma class GST Chain A, Glucose-6-phosphate isomerase, pyruvate dehydrogenase subunit E1, Glutamate dehydrogenase 2, arachin Ahy-1, Full = Allergen Ara h 1, clone P41B, Gly1. The data generated in this study show that there is sharing of antigens between organisms of different taxonomic groups, but also with allergens tested. Moreover, the description and analysis of shared molecular components can help in understanding the evolutionary history and phylogeny of these organisms. / Duas esp?cies de parasitos nemat?deos da fam?lia Angiostrongylidae, do g?nero Angiostrongylus de localiza??o intra-arterial s?o capazes de produzir doen?a em humanos: Angiostrongylus costaricensis e Angiostrongylus cantonensis. Ambas as esp?cies s?o parasitos pr?prios de roedores e a infec??o humana ? considerada acidental. No homem, A. cantonensis ? o causador da meningite eosinof?lica e A. costaricensis causador da angiostrongil?ase abdominal. Como n?o h? elimina??o de formas parasit?rias na infec??o humana, o diagn?stico se torna dif?cil e m?todos moleculares se tornam necess?rios. Depois de anos da descoberta das angiostrongil?ases, ainda h? muitos esfor?os no estudo e desenvolvimento de um teste de diagn?stico espec?fico e sens?vel capaz de discriminar as Angiostrongil?ases de outras parasitoses. Neste contexto, a reatividade cruzada se torna um problema na especificidade de testes sorol?gicos. O objetivo principal deste trabalho foi analisar o compartilhamento de ant?genos entre Angiostrongylus spp. e os parasitos Strongyloides spp., Fasciola hepatica, Ascaris lumbricoides, Hymenolepis diminuta e Toxocara canis, assim como identificar essas mol?culas compartilhadas. Para obten??o de ant?genos, algumas ratazanas foram capturas e suas fezes analisadas atrav?s do m?todo de Baerman al?m de terem sido semeadas em Placas de ?gar. Foram obtidas larvas de Strongyloides spp. e Angiostrongylus spp. Nas art?rias pulmonares de uma ratazana foram encontrados 11 vermes f?meas e 2 vermes machos de A. cantonensis, que deu origem ao primeiro relato da ocorr?ncia deste parasito no Rio Grande do Sul. No intestino delgado foi obtido um verme de Strongyloides spp e da an?lise do intestino delgado foram obtidos vermes de Hymenolepis diminuta. A partir dos vermes adultos dos parasitos, foram obtidos os ant?genos utilizados neste trabalho. Al?m disso, foram utilizados ant?genos de alergenos como o amendoim, tomate, p?len e morango. Para identifica??o dos ant?genos compartilhados, as prote?nas foram separadas por eletroforese uni e bidimensional ensaiadas pela t?cnica de Western-Blot utilizando-se soro de pacientes com angiostrongil?ases. Todos os ant?genos foram reconhecidos pelo soro de pacientes infectados com angiostrongil?ases, sendo um total de quatorze bandas identificadas como imunog?nicas. As quartorze bandas foram recortadas dos g?is de poliacrilamida e analisadas por espectrometria de massas. As prote?nas identificadas foram: Fosfoenolpiruvato carboxiquinase, Prote?na de choque t?rmico 70 (HSP70), Prote?na hipot?tica DAPPUDRAFT, 60 kDa Chaperonina 5, Gliceralde?do-3-fosfato-desidrogenase, Cadeia A sigma classe GST, Glucose-6-fosfato isomerase, Piruvato desidrogenase subunidade E1, Glutamato desidrogenase 2 , arachin Ahy-1, Full=Allergen Ara h 1, clone P41B , Gly1. Os dados gerados no presente trabalho demonstram que h? compartilhamento de ant?genos entre organismos de diferentes grupos taxon?micos, como tamb?m com os alergenos testados. Al?m disso, a descri??o e a an?lise de componentes moleculares compartilhados podem ajudar na compreens?o da hist?ria evolutiva e da filogenia destes organismos.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/250 |
| Date | 28 February 2013 |
| Creators | Cognato, Bianca Barbieri |
| Contributors | Graeff-teixeira, Carlos |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Zoologia, PUCRS, BR, Faculdade de Bioci?ncias |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 2008925231902741151, 500, 600, 36528317262667714 |
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