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Previous issue date: 2008-02-22 / Funda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Embryonic cells of tick in vitro constitute an important one tool for culture and study of the
biology of B. burgdorferi. Spirochetes Borrelia burgdorferi is the aetiologic agent of borreliose
of Lyme in U.S.A. and Europe, where it is transmitted by tick of the Ixodes genus. The aim of
this work was in vitro to cultivate B. burgdorferi (American Cepa G39/40) in primary culture of
embryonic cells of Rhipicephalus (B.) microplus and the A. cajennense. From the primary culture
of embryonic cells of R. (B.) microplus were performed subculture maintained with medium
Leibovitz's (L-15) free of antibiotic, that later it were changed by medium Barbour-Stoener-Kelly
(BSK). After the addition of the BSK inoculated B. burgdorferi of and also in Tube of Leighton
(TL) with free BSK of cells (control) in a final concentration of 6x106 spirochetes/mL. The
embryonic cells of the A. cajennense initially were cultivated in medium L-15 with antibiotic,
which was substituted by the BSK. Later, 1,1x107 spirochetes/mL in medium BSK cultivated was
inoculated, and also in TL controlled free of cells. All the culture was incubated at 34?C. The
development of the culture was observed in microscope of inverted contrast of phase, as well as
the countings of B. burgdorferi performed in chamber of neubauer. Cover glass of the TL had
been stained with Giemsa. It was evidenced by the observation in microscope of inverted contrast
of phase the survival, attach and multiplication of B. burgdorferi, in the embryonic cells of R. (B.)
microplus and A. cajennense. It did not have differences in the finale counting of spirochetes
cultivated in cells of R. (B.) microplus when compared with the free control of cells, but on with
cells of the A. cajennense, the number amount approximately was 1,9x107 spirochetes/mL, and
while in the tube it has controlled free of cells was 1x106 spirochetes /mL. The culture of cells of
tick R. (B.) microplus and the A. cajennense have potential to be used as substratum for culture of
B. burgdorferi, and study of its development. / C?lulas embrion?rias de carrapatos mantidas in vitro constituem uma importante ferramenta para
cultivo e estudo da biologia de Borrelia burgdorferi. A espiroquetas B. burgdorferi ? o agente
etiol?gico da borreliose de Lyme nos EUA e Europa, onde ? transmitida por carrapatos do g?nero
Ixodes. O objetivo deste trabalho foi cultivar in vitro Borrelia burgdorferi (Cepa Americana
G39/40) em cultura prim?ria de c?lulas embrion?rias dos carrapatos Rhipicephalus (Boophilus)
microplus e Amblyomma cajennense. A partir da cultura prim?ria de c?lulas embrion?rias de R.
(Boophilus) microplus foram realizados subcultivos mantidos com meio Leibovitz s L-15 livre de
antibi?tico, que posteriomente foi trocado pelo meio Barbour-Stoener-Kelly (BSK). Ap?s a
adi??o do BSK na cultura, inoculou-se B. burgdorferi e tamb?m em Tubo de Leighton (TL) com
BSK livres de c?lulas (controle), numa concentra??o final de 6x106 espiroquetas/mL. As c?lulas
embrion?rias de A. cajennense foram inicialmente cultivadas em meio L-15 com antibi?tico, o
qual foi substitu?do pelo BSK. Posteriormente, inoculou-se 1,1x107 espiroquetas/mL cultivadas
em meio BSK, e tamb?m em TL controle livre de c?lulas. Todos os cultivos foram incubados em
estufa bacteriol?gica a 34?C. O desenvolvimento dos cultivos foram observados em microsc?pio
de contraste de fase invertido, assim como as contagens de B. burgdorferi realizados em c?mara
de neubauer. As lam?nulas dos TL foram coradas com Giemsa. Foi constatado pela observa??o
em microsc?pio de contraste de fase invertido a sobreviv?ncia, ader?ncia e multiplica??o de B.
burgdorferi, nas c?lulas embrion?rias de R. (Boophilus) microplus e A. cajennense. N?o houve
diferen?as na contagem final de espiroquetas cultivadas em c?lulas de R. (Boophilus) microplus
quando comparada ao controle livre de c?lulas, mas sobre c?lulas de A. cajennense o valor total
de aproximadamente 1,9x107 espiroquetas/mL, e enquanto no tubo controle livre de c?lulas foi
1x106 espiroquetas/mL. O cultivo de c?lulas do carrapato R. (Boophilus) microplus e A.
cajennense t?m potencial para ser utilizado como substrato para cultivo de B. burgdorferi e para
estudo de seu desenvolvimento.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:tede/759 |
| Date | 22 February 2008 |
| Creators | Rezende, Jania de |
| Contributors | Fonseca, Adivaldo Henrique da |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Curso de P?s-Gradua??o em Ci?ncias Veterin?rias, UFRRJ, Brasil, Parasitologia Veterin?ria |
| 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 UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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