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An?lise do crescimento, diferencia??o celular e perfil isoenzim?tico de um isolado de tripanosomat?deo obtido de Megaselia scalaris Loew, 1866 (D?ptera, Phoridae) / Analysis of growth, cell differentiation and isoenzyme profile of an isolated trypanosomatid obtained Megaselia scalaris Loew, 1866 (Diptera, Phoridae)Lima , Nathanielly Rocha Casado de 30 March 2011 (has links)
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Previous issue date: 2011-03-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The Trypanosomatidae family includes parasites of a wide variety of vertebrates, invertebrates (mainly insects), plants and some species of protozoa. In this study, an isolate was obtained by feces culturing of the intestinal tract of Megaselia scalaris Loew, 1866 (Diptera, Phoridae). The original isolate and its clones (obtained by flow cytometry) were deposited in the ?Trypanosomatids Collection of the Instituto Oswaldo Cruz. These clones were characterized by different approaches I in comparison with the reference species of different genera. Its growth has been studied in LIT medium (with 10 or 20% fetal bovine serum) modified according to the requirement. The number of cells per microliter was estimated at 24 h intervals between 48-144 h. Giemsa-stained smears (after acid hydrolysis) were prepared in order to study cell differentiation and morphology of the forms. The isozyme analysis was performed using the following systems: GPI, PGM, 6PGDH, ME, IDH, MDH, ACON, FUM, MPI and HK. Data from this analysis were processed and analyzed numerically using the computacional Jaccard coefficient and UPGMA algorighm. The result set of this study suggest that: (1) The strain of M. scalaris showed 70% similarity with Herpetomonas megaseliae, (2) This isolate showed the exclusive presence of promastigotes in any analysis of the growth curve as expected for the genus Leptomonas, however the forms exhibited high polymorphism. Additional studies using molecular tools will be realized / A fam?lia Trypanosomatidae inclui parasitas de uma grande variedade de vertebrados, invertebrados (principalmente insetos), plantas e algumas esp?cies de protozo?rios. No presente trabalho, um isolado foi obtido por coprocultivo de trato intestinal de Megaselia scalaris Loew, 1866 (Diptera, Phoridae). O isolado original e seus clones (obtidos por citometria de fluxo) foram depositados na "Cole??o de Tripanosomat?deos do Instituto Oswaldo Cruz". Estes clones foram caracterizados por diversas abordagens em compara??o com esp?cies de refer?ncia de diferentes g?neros. Seu crescimento foi estudado em meio LIT (com 10 ou 20% de soro fetal bovino) modificado, de acordo com o n?vel de exig?ncia. O n?mero de c?lulas por microlitro foi estimado em intervalos de 24 h, entre 48-144 h. Esfrega?os corados pelo Giemsa (ap?s hidr?lise ?cida) foram preparados visando estudos de diferencia??o celular e morfometria das formas encontradas. A an?lise de isoenzimas foi realizada utilizando-se os seguintes sistemas: GPI , PGM, 6PGDH, ME, IDH, MDH, ACON, FUM, MPI e HK. Os dados desta an?lise foram processados numericamente e submetidos ? an?lise computacional utilizando-se o coeficiente de Jaccard e o algoritmo UPGMA. O conjunto de resultados deste trabalho sugere que: (1) O isolado de M. scalaris apresentou similaridade de 70% com Herptomonas megaseliae; (2) Esse isolado revelou a presen?a exclusiva de promastigotas em toda an?lise da curva de crescimento, como esperado para o g?nero Leptomonas, entretanto as formas exibiram grande polimorfismo. Estudos complementares utilizando ferramentas moleculares ser?o realizados.
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