Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)

Ba?ta, Bruna de Azevedo

Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)

Detalhes bibliográficos
Ano de defesa:	2011
Autor(a) principal:	Ba?ta, Bruna de Azevedo
Orientador(a):	Fonseca, Adivaldo Henrique da
Banca de defesa:	Ribeiro, M?cio Fl?vio Barbosa, Oliveira, ?ngela de, Cunha, Nathalie Costa da
Tipo de documento:	Dissertação
Tipo de acesso:	Acesso aberto
Idioma:	por
Instituição de defesa:	Universidade Federal Rural do Rio de Janeiro
Programa de Pós-Graduação:	Programa de P?s-Gradua??o em Ci?ncias Veterin?rias
Departamento:	Instituto de Veterin?ria
País:	Brasil
Palavras-chave em Português:	carrapato espiroquetas cultivo celular
Palavras-chave em Inglês:	ticks spirochetes cell culture
Área do conhecimento CNPq:	Medicina Veterin?ria
Link de acesso:	https://tede.ufrrj.br/jspui/handle/jspui/3908
Resumo:	The aims of this study were to establish the primary culture of embryonic cells from Dermacentor nitens and evaluate the influence of the Barbour-Stoenner-Kelly (BSK) and Leibovitz's L-15B media in co-culture with Borrelia burgdorferi (American strain G39/40). The culture was established from embryonated eggs of ingurgitated females of D. nitens with 13 days of laying, using the Leibovitz's L-15B medium supplemented. After the formation of monolayer in flasks of 10cm2, the L-15B medium was removed from the flasks. Three groups with different media were formed. A group consisting only of BSK medium (group I), a group with L-15B medium with 40% BSK (group II) and a third group with L-15B medium with 10% BSK (group III). For each group three repetitions were done, which received the spirochetes inoculum of B. burgdorferi, with final concentration of approximately 1.1 x 106 spirochetes/mL. It was prepared another flask without tick cells with 5 mL of BSK, in order to assess the development of spirochetes in the absence of embryonic cells. The count of B. burgdorferi was performed three days after inoculation of spirochetes. Since the second day of culture it was observed fixation of most cells on the surface of the flask, with numerous cell aggregates. After fixation, it was observed a wide variety of cell types that began to differentiate into fibroblastoid cells and posteriorly they start to differentiate into epithelioid and rounded cells. There was extensive proliferation of spirochetes cultured with embryonic cells in comparison to the initial concentration. Most spirochetes was epicellular and was attached to the cells longitudinally or by their edges, with few free spirochetes. Group II showed the best results, since the medium caused less damage to tick cells in comparison to Group I with good multiplication of spirochetes in comparison to groups III and control. Although D. nitens is not species-specific vector of B. burgdorferi, the spirochete culture in this study was successful, proving to be a useful tool in the isolation of strains or species of Borrelia spp.

Metadados do item

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spelling	Fonseca, Adivaldo Henrique da475.018.557-49http://lattes.cnpq.br/4411441162862608Ribeiro, M?cio Fl?vio BarbosaOliveira, ?ngela deCunha, Nathalie Costa da101.354.697-08http://lattes.cnpq.br/8016012971743463Ba?ta, Bruna de Azevedo2020-09-18T14:24:00Z2011-02-17BA?TA, Bruna de Azevedo. Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae). 2011. 8 f. Disserta??o( Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.https://tede.ufrrj.br/jspui/handle/jspui/3908The aims of this study were to establish the primary culture of embryonic cells from Dermacentor nitens and evaluate the influence of the Barbour-Stoenner-Kelly (BSK) and Leibovitz's L-15B media in co-culture with Borrelia burgdorferi (American strain G39/40). The culture was established from embryonated eggs of ingurgitated females of D. nitens with 13 days of laying, using the Leibovitz's L-15B medium supplemented. After the formation of monolayer in flasks of 10cm2, the L-15B medium was removed from the flasks. Three groups with different media were formed. A group consisting only of BSK medium (group I), a group with L-15B medium with 40% BSK (group II) and a third group with L-15B medium with 10% BSK (group III). For each group three repetitions were done, which received the spirochetes inoculum of B. burgdorferi, with final concentration of approximately 1.1 x 106 spirochetes/mL. It was prepared another flask without tick cells with 5 mL of BSK, in order to assess the development of spirochetes in the absence of embryonic cells. The count of B. burgdorferi was performed three days after inoculation of spirochetes. Since the second day of culture it was observed fixation of most cells on the surface of the flask, with numerous cell aggregates. After fixation, it was observed a wide variety of cell types that began to differentiate into fibroblastoid cells and posteriorly they start to differentiate into epithelioid and rounded cells. There was extensive proliferation of spirochetes cultured with embryonic cells in comparison to the initial concentration. Most spirochetes was epicellular and was attached to the cells longitudinally or by their edges, with few free spirochetes. Group II showed the best results, since the medium caused less damage to tick cells in comparison to Group I with good multiplication of spirochetes in comparison to groups III and control. Although D. nitens is not species-specific vector of B. burgdorferi, the spirochete culture in this study was successful, proving to be a useful tool in the isolation of strains or species of Borrelia spp.Os objetivos do presente estudo foram estabelecer o cultivo prim?rio de c?lulas embrion?rias de Dermacentor nitens e avaliar a influ?ncia dos meios Barbour-Stoenner-Kelly (BSK) e Leibovitz?s L-15B no co-cultivo com Borrelia burgdorferi (cepa americana G39/40). A cultura foi estabelecida a partir de ovos embrionados de f?meas ingurgitadas de D. nitens com 13 dias ap?s o in?cio da postura, utilizando o meio de cultivo Leibovitz?s L-15B suplementado. Ap?s a forma??o da monocamada nos frascos de 10cm2, o meio de cultura L-15B foi retirado dos tubos. Foram formados tr?s grupos com meios distintos, um grupo com meio constitu?do apenas de BSK (grupo I), um grupo com meio L-15B com 40% de BSK (grupo II) e um terceiro grupo com meio L-15B com 10% de BSK (grupo III). Para cada grupo foram realizadas tr?s repeti??es, as quais receberam os in?culos de espiroquetas de B. burgdorferi, apresentando concentra??o final de aproximadamente 1,1 x 106 espiroquetas/mL. Foi preparado mais um tubo sem c?lulas de carrapato com 5mL de BSK, com a finalidade de avaliar o desenvolvimento das espiroquetas na aus?ncia de c?lulas embrion?rias. A contagem de B. burgdorferi foi realizada tr?s dias ap?s a inocula??o das espiroquetas. A partir do segundo dia de in?cio de cultivo foi observada a fixa??o da maioria das c?lulas na superf?cie do frasco, com in?meros agregados celulares. Ap?s fixa??o, foi observada uma grande variedade de tipos celulares que come?aram a se diferenciar em c?lulas fibroblast?ides e posteriormente, c?lulas epiteli?ides e arredondadas. Houve grande multiplica??o das espiroquetas cultivadas com c?lulas embrion?rias quando comparada ? concentra??o inicial. A maioria das espiroquetas se apresentava epicelular e estava aderida ?s c?lulas longitudinalmente ou pelas suas extremidades, com poucas espiroquetas livres. O grupo II, demonstrou melhores resultados, visto que, os meios causaram menores danos ?s c?lulas de carrapato quando comparados ao grupo I e com boa multiplica??o de espiroquetas quando comparados aos grupos III e controle. Embora D. nitens n?o seja esp?cie vetora espec?fica de B. burgdorferi, o cultivo da espiroqueta no estudo foi bem sucedido, demonstrando ser uma ferramenta ?til na tentativa de isolamento de cepas ou esp?cies de Borrelia spp.Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2020-09-18T14:24:00Z No. of bitstreams: 1 2011 - Bruna de Azevedo Ba?ta.pdf: 855803 bytes, checksum: 0acd395ed803e42d6d3a9110df4fe74b (MD5)Made available in DSpace on 2020-09-18T14:24:00Z (GMT). No. of bitstreams: 1 2011 - Bruna de Azevedo Ba?ta.pdf: 855803 bytes, checksum: 0acd395ed803e42d6d3a9110df4fe74b (MD5) Previous issue date: 2011-02-17Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior, CAPES, Brasil.application/pdfhttps://tede.ufrrj.br/retrieve/62023/2011%20-%20Bruna%20de%20Azevedo%20Ba%c3%aata.pdf.jpgporUniversidade Federal Rural do Rio de JaneiroPrograma de P?s-Gradua??o em Ci?ncias Veterin?riasUFRRJBrasilInstituto de Veterin?riaABELE, D. C.; ANDER, K. H. The many faces and phases of borreliosis I. Lyme Disease. Journal of the American Academy of Dermatology, v. 23, n. 2, p. 167-186, 1990. ARAG?O, H. B. Ixodidas brasileiros e de alguns pa?ses limitadores. Mem?ria do Instituto Oswaldo Cruz, v. 31, n.4, p. 759-844, 1936. BALASHOV, Y. S. Bloodsucking Insects and Ticks and Mites, Vectors of transmissible Infections of Humans and Domestic Animals. Entomological Review, v. 85, n. 8, p. 990-1007, 2005. BARBOUR, A. G.; HAYES, S. F. Biology of Borrelia species. 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Revista Hospital das Cl?nicas da Faculdade de Medicina de S?o Paulo, v. 52, n. 2, p. 111-117, 1997. YOSHINARI, N. H.; BARROS, P. J. L.; FONSECA, A. H.; BONOLDI, V. L. N.; BARROS-BATTESTI, D. M.; SCHUMAKER, T. S.; COSSERMELLI, W. Borreliose de Lyme - Zoonose emergente de interesse multidisciplinar. News Laboratorial, v. 3, n.12, p. 90-104, 1995. YOSHINARI, N. H.; BARROS, P. J. L.; YASSUDA, P.; BAGGIO, D.; STEERE, A. C.; PAGLIARINE, R. C.; COSSERMELLI, W. Estudo epidemiol?gico da doen?a de Lyme no Brasil. Revista Hospital das Cl?nicas da Faculdade de Medicina de S?o Paulo, v. 47, n. 2, p. 71-75, 1992. YOSHINARI, N. H.; MANTOVANI, E.; BONOLDI, V. L. N.; MARANGONI, R. G.;GAUDITANO, G. Doen?a de lyme-s?mile brasileira ou s?ndrome Baggio-Yoshinari: zoonose ex?tica e emergente transmitida por carrapatos. Revista da Associa??o M?dica Brasileira. v. 56, n.3, p. 363-369, 2010. 28 YOSHINARI, N. H.; OYAFUSO, L. K.; MONTEIRO, F. G. V.; BARROS, P. J. L.; CRUZ, F. C. M.; FERREIRA, L. G. E.; BONASSER, F.; BAGGIO, D.; COSSERMELLI, W. Doen?a de Lyme: Relato de um caso observado no Brasil. Revista Hospital das Cl?nicas da Faculdade de Medicina de S?o Paulo, v. 48, n. 4, p.170-174, 1993. YUNKER, C. E. Preparation and maintenance of arthropod cell cultures: Acari, with emphasis on ticks. In: YUNKER, C. E. Arboviruses in arthropod cells in vitro. Boca Raton: CRC Press, 1987. p. 35-51. YUNKER, C. E.; CORY, J. Effectiveness of refrigerated nymphs in tick tissue culture experiments. Journal of Parasitology, v. 51, n. 4, p. 686, 1965. ZUNG, J. L.; LEWENGRUB, S.; RUDZINSKA, M. A.; SPIELMAN, A.; TELFORD, A. R.; PIESMAN, J. Fine structural evidence for the penetration of the Lyme disease spirochete Borrelia burgdoferi through the gut and salivary tissues of lxodes dammini. 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dc.title.por.fl_str_mv	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
dc.title.alternative.eng.fl_str_mv	Co-culture of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) in primary culture of embryonic cells from Dermacentor nitens (Acari: Ixodidae)
title	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
spellingShingle	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae) Ba?ta, Bruna de Azevedo carrapato espiroquetas cultivo celular ticks spirochetes cell culture Medicina Veterin?ria
title_short	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
title_full	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
title_fullStr	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
title_full_unstemmed	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
title_sort	Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)
author	Ba?ta, Bruna de Azevedo
author_facet	Ba?ta, Bruna de Azevedo
author_role	author
dc.contributor.advisor1.fl_str_mv	Fonseca, Adivaldo Henrique da
dc.contributor.advisor1ID.fl_str_mv	475.018.557-49
dc.contributor.advisor1Lattes.fl_str_mv	http://lattes.cnpq.br/4411441162862608
dc.contributor.referee1.fl_str_mv	Ribeiro, M?cio Fl?vio Barbosa
dc.contributor.referee2.fl_str_mv	Oliveira, ?ngela de
dc.contributor.referee3.fl_str_mv	Cunha, Nathalie Costa da
dc.contributor.authorID.fl_str_mv	101.354.697-08
dc.contributor.authorLattes.fl_str_mv	http://lattes.cnpq.br/8016012971743463
dc.contributor.author.fl_str_mv	Ba?ta, Bruna de Azevedo
contributor_str_mv	Fonseca, Adivaldo Henrique da Ribeiro, M?cio Fl?vio Barbosa Oliveira, ?ngela de Cunha, Nathalie Costa da
dc.subject.por.fl_str_mv	carrapato espiroquetas cultivo celular
topic	carrapato espiroquetas cultivo celular ticks spirochetes cell culture Medicina Veterin?ria
dc.subject.eng.fl_str_mv	ticks spirochetes cell culture
dc.subject.cnpq.fl_str_mv	Medicina Veterin?ria
description	The aims of this study were to establish the primary culture of embryonic cells from Dermacentor nitens and evaluate the influence of the Barbour-Stoenner-Kelly (BSK) and Leibovitz's L-15B media in co-culture with Borrelia burgdorferi (American strain G39/40). The culture was established from embryonated eggs of ingurgitated females of D. nitens with 13 days of laying, using the Leibovitz's L-15B medium supplemented. After the formation of monolayer in flasks of 10cm2, the L-15B medium was removed from the flasks. Three groups with different media were formed. A group consisting only of BSK medium (group I), a group with L-15B medium with 40% BSK (group II) and a third group with L-15B medium with 10% BSK (group III). For each group three repetitions were done, which received the spirochetes inoculum of B. burgdorferi, with final concentration of approximately 1.1 x 106 spirochetes/mL. It was prepared another flask without tick cells with 5 mL of BSK, in order to assess the development of spirochetes in the absence of embryonic cells. The count of B. burgdorferi was performed three days after inoculation of spirochetes. Since the second day of culture it was observed fixation of most cells on the surface of the flask, with numerous cell aggregates. After fixation, it was observed a wide variety of cell types that began to differentiate into fibroblastoid cells and posteriorly they start to differentiate into epithelioid and rounded cells. There was extensive proliferation of spirochetes cultured with embryonic cells in comparison to the initial concentration. Most spirochetes was epicellular and was attached to the cells longitudinally or by their edges, with few free spirochetes. Group II showed the best results, since the medium caused less damage to tick cells in comparison to Group I with good multiplication of spirochetes in comparison to groups III and control. Although D. nitens is not species-specific vector of B. burgdorferi, the spirochete culture in this study was successful, proving to be a useful tool in the isolation of strains or species of Borrelia spp.
publishDate	2011
dc.date.issued.fl_str_mv	2011-02-17
dc.date.accessioned.fl_str_mv	2020-09-18T14:24:00Z
dc.type.status.fl_str_mv	info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv	info:eu-repo/semantics/masterThesis
format	masterThesis
status_str	publishedVersion
dc.identifier.citation.fl_str_mv	BA?TA, Bruna de Azevedo. Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae). 2011. 8 f. Disserta??o( Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
dc.identifier.uri.fl_str_mv	https://tede.ufrrj.br/jspui/handle/jspui/3908
identifier_str_mv	BA?TA, Bruna de Azevedo. Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae). 2011. 8 f. Disserta??o( Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
url	https://tede.ufrrj.br/jspui/handle/jspui/3908
dc.language.iso.fl_str_mv	por
language	por
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Carrapatos de Import?ncia M?dico-Veterin?ria da Regi?o Neotropical: Um guia ilustrado para identifica??o de esp?cies, Vox/ICTTD-3/Butantan, 2006, 223 p. BARROS-BATTESTI, D. M.; YOSHINARI, N. H.; BONOLDI, V. L. N.; GOMES, A. C. Parasitism by Ixodes didelphidis and I. loricatus (Acari: Ixodidae) on Small Wild Mammals from an Atlantic Forest in the State of Sao Paulo, Brazil. Journal of Medical Entomology., v. 37, n. 6, p. 820-827, 2000. BECK, G.; HABICHT, G. S; BENACH, J. L.; COLEMAN, J. L. Chemical and biologic characterization of a lipopolysaccharide extracted from the Lyme disease spirochete (Borrelia burgdorferi) Journal of infectious diseases, v. 152, n. 1, p.108-117, 1985 BELL- SAKYI, L.; PAXTON, E. A.; MUNDERLOH, U. G.; SUMPTION, K. J. Growth of ruminatium, the causative agent of heartwater, in a tick line. Journal of Clinical Microbiology v. 38, n. 3, p. 1238-1240, 2000. BELL-SAKYI, L.; ZWEYGARTH, E.; BLOUIN, E. F.; GOULD, E. A.; JONGEJAN, F. Tick cell lines: tools for tick and tick-borne disease research. Trends in Parasitology, v. 23, n. 9, p. 450-457, 2007. BENACH, J. L.; COLEMAN, J. H.; SKINNER, R. A.; BOSLER, E. M. Adult lxodes dammini on rabbits: a hypothesis for the development and transmission of Borrelia burgdorferi. The Journal of Infectious Diseases, v. 155, n. 6, p. 1300-1306, 1987. BENNETT, C. E. Ticks and Lyme disease. Advances in Parasitology, v. 36, n. 1, p. 343-405, 1995. 21 BENXIU, J.; THOMAS, C. B.; COLLINS, M. T. Evaluation of an enzyme-linked immunosorbent assay that uses the 41-kd flagellin as the antigen for detection of antibodies to Borrelia burgdorferi in cattle. American Journal of Veterinary Research, v. 55, n. 9, p. 1213-1219, 1994. BLOUIN, E. F.; DE LA FUENTE, J.; GARCIA-GARCIA, J. C.; SAUER, J. R.; SALIKI, J. T.; KOCAN, K. M. Applications of a cell culture system for studying the interaction of Anaplasma marginale with tick cells. Animal Health Research Reviews, v. 3, n. 2, p. 57-68, 2002. 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Co-cultivo de Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) em cultura prim?ria de c?lulas embrion?rias de Dermacentor nitens (Acari: Ixodidae)

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