Thesis (Ph. D.)--University of Chicago, Dept. of Ecology and Evolution. / Includes bibliographical references. Also available on the Internet.

Thesis (Ph.D.) -- University of Rhode Island, 2009. / Typescript. Includes bibliographical references (leaves 188-202).

Thesis (M.A.) -- Central Connecticut State University, 2006. / Thesis advisor: Kathy Martin. "... in partial fulfillment of the requirements for the degree of Master of Arts in Biomolecular Sciences." Includes bibliographical references (leaves 99-103). Also available via the World Wide Web.

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Thesis (Ph. D.)--West Virginia University, 2005 / Title from document title page. Document formatted into pages; contains ix, 210 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-07-21T12:10:16Z No. of bitstreams: 1 2010 - Rafaella Camara Teixeira.pdf: 2127524 bytes, checksum: 2f2281ce314ac38db40bc03d628e7447 (MD5) / Made available in DSpace on 2017-07-21T12:10:16Z (GMT). No. of bitstreams: 1 2010 - Rafaella Camara Teixeira.pdf: 2127524 bytes, checksum: 2f2281ce314ac38db40bc03d628e7447 (MD5) Previous issue date: 2010-02-25 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq, Brasil. / Cell cultures provide a simplified system of observation that can be particularly useful for studies of intracellular and epicellular microorganisms. The aim of this study was to establish in vitro embryonic cells primary culture of the tick Rhipicephalus sanguineus to cultivate the spirochete Borrelia burgdorferi american strain G39/40. The culture was established from embryonated eggs of engorged females of R. sanguineus to 12 days after the beginning of the oviposition, using the culture medium Leibovitz's L-15B supplemented with 20% of inactivated fetal calf serum, 10% of tryptose phosphate broth, 0.1% fraction V bovine albumin, 1% of glutamine and 0.1% of gentamicin antibiotic, pH 6.8. After the formation of a monolayer, the initial culture medium L-15B was removed from the tubes and replaced by Barbour-Stoenner-Kelly medium (BSK) or BSK with L-15B without antibiotics. Spirochetes previously grown in BSK were counted and inoculated into tubes, with final concentration of approximately 6.2 x 105 spirochetes/mL. B. burgdorferi from the inoculated tubes were countered when the means showed yellow color, indicative of high acidity due to the multiplication of spirochetes. On the third day after the start of primary culture of R. sanguineus embryonic cells, we observed the fixation of cell aggregates on the surface of the bottles. From these clusters, there were several cell types, such as large fibroblast-type cells and structures like vesicles and tubes. In the second week, we observed the appearance of round or flattened epithelial-type cells, and after 21 days of culture, we realized the formation of a monolayer due to the appearance of confluent cells. The L-15B medium proved to be efficient for the development of primary culture of R. sanguineus embryonic cells. There was a great multiplication of spirochetes cultivated with cultured embryonic cells when compared to the initial concentration, as well as the spirochetes grown in the absence of the tick cells, observing an increase of 100 times the number of B. burgdorferi. Seven days after inoculation, the tubes in which we used only the BSK medium, higher concentrations of B. burgdorferi were recovered when compared to the tubes where the medium BSK and Leibovitz's L-15B were used. Regardless of the culture media tested, the final concentration of B. burgdorferi of the tubes with embryonic tick cells was lower than that of seamless embryonic cells. In observation of the culture tubes on microscopy phase contrast, spirochetes were presented adhered to epithelial-type and fibroblast-type tick cells in an epicelular way and with great motility. R. sanguineus embryonic cells grown in BSK medium, with or without B. burgdorferi inoculation, stopped its propagation, showed membrane degeneration and many of them broke away from the surface of the bottle. The cells grown in BSK and L- 15B continued to multiply, many were still intact and attached to the bottle, with the presence of tissues in development, with fewer degenerated and floating cells than those cultivated in BSK. The spirochete B. burgdorferi strain G39/40, adhered, grew, multiplied and showed great motility in cultures of embryonic cells of R. sanguineus tick, using BSK and Leibovitz?s L-15B media. / As culturas celulares oferecem um simplificado sistema de observa??o que pode ser particularmente ?til para estudos de microrganismos intracelulares e epicelulares. O objetivo deste estudo foi estabelecer cultura prim?ria in vitro de c?lulas embrion?rias do carrapato Rhipicephalus sanguineus para cultivo da espiroqueta Borrelia burgdorferi, cepa americana G39/40. A cultura foi estabelecida a partir de ovos embrionados de f?meas ingurgitadas de R. sanguineus com 12 dias ap?s o ?nicio da postura, utilizando o meio de cultivo Leibovitz?s L- 15B, suplementado com 20% de soro fetal bovino inativado, 10% de caldo triptose fosfato, 0,1% fra??o V de albumina bovina, 1% de glutamina e 0,1% de antibi?tico gentamicina, pH 6,8. Com a forma??o de uma monocamada celular, o meio de cultura inicial L-15B foi retirado dos tubos e trocado por meio Barbour-Stoenner-Kelly (BSK) ou BSK com L-15B sem antibi?tico. As espiroquetas previamente cultivadas em BSK foram contadas e inoculadas nos tubos, apresentando concentra??o final de aproximadamente 6,2 x 105 espiroquetas/mL. A contagem de B. burgdorferi dos tubos inoculados foi realizada quando o meio apresentou colora??o amarela, indicativa de elevada acidez devido ? multiplica??o das espiroquetas. No terceiro dia ap?s o in?cio da cultura prim?ria de c?lulas embrion?rias de R. sanguineus, foi poss?vel observar a fixa??o de agregados celulares na superf?cie dos frascos. A partir destes agregados, surgiram diversos tipos celulares, como grandes c?lulas fibroblast?ides e estruturas semelhantes a ves?culas e tubos. Na segunda semana, foi observado o aparecimento das c?lulas epiteli?ides ou redondas e, com 21 dias de cultivo, visualizou-se a forma??o de uma monocamada celular devido ao aspecto confluente das c?lulas. O meio de cultivo L-15B demonstrou ser eficiente para o desenvolvimento da cultura prim?ria de c?lulas embrion?rias de R. sanguineus. Houve grande multiplica??o das espiroquetas cultivadas com c?lulas embrion?rias quando comparada ? concentra??o inicial, assim como das espiroquetas cultivadas na aus?ncia das c?lulas de carrapato, observando-se um aumento em 100 vezes do n?mero de B. burgdorferi. Sete dias ap?s a inocula??o, foram recuperadas maiores concentra??es de B. burgdorferi nos tubos onde se utilizou somente o meio BSK, do que nos tubos onde foi utilizado BSK juntamente com Leibovitz?s L-15B. Independente dos meios de cultivo testados, a concentra??o final de B. burgdorferi dos tubos com c?lulas embrion?rias de carrapato foi menor do que a dos tubos sem c?lulas embrion?rias. Na observa??o dos tubos de cultivo ? microscopia de contraste de fase, as espiroquetas apresentaram-se aderidas ?s c?lulas de carrapato epiteli?ides e fibroblast?ides de maneira epicelular e com grande motilidade. As c?lulas embrion?rias de R. sanguineus cultivadas em meio BSK, com ou sem in?culo de B. burgdorferi, pararam sua multiplica??o, apresentaram degenera??o na membrana e muitas desprenderam-se da superf?cie do frasco. As c?lulas cultivadas em meio BSK e L-15B continuaram a se multiplicar, muitas ainda estavam ?ntegras e aderidas ao frasco, com presen?a de tecidos em desenvolvimento, com menos c?lulas degeneradas e flutuantes que as cultivadas somente em BSK. A espiroqueta B. burgdorferi, cepa G39/40, aderiu, cresceu, multiplicou e apresentou grande motilidade nos cultivos com c?lulas embrion?rias do carrapato R. sanguineus, utilizando meios BSK e Leibovitz?s L-15B.

Thesis (M.S.)--Ball State University, 2009. / Title from PDF t.p. (viewed on Feb. 08, 2010). Includes bibliographical references (p. 81-94).

Thesis--Auburn University, 2005. / Abstract. Vita. Includes bibliographic references.

Thesis (M.S.)--University of Tennessee, Knoxville, 2009. / Title from title page screen (viewed on Mar. 18, 2010). Thesis advisor: Graham Hickling. Vita. Includes bibliographical references.

Thesis (Ph. D.)--University of Missouri-Columbia, 1999. / Typescript. Vita. Includes bibliographical references (leaves 70-85). Also available on the Internet.