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dc.contributor.authorOliveira, Ângela de
dc.date.accessioned2023-12-21T18:43:43Z-
dc.date.available2023-12-21T18:43:43Z-
dc.date.issued2002-03-21
dc.identifier.citationOLIVEIRA, Ângela de. Cultivo de Borrelia burgdorferi e B. garinii (Spirochaetales:Spirochaetaceae): cinética de crescimento em diferentes meios de cultivo e técnicas de coloração. 2002. 113 f. Tese (Doutorado em Ciências Veterinárias) - Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica - RJ, 2002.por
dc.identifier.urihttps://rima.ufrrj.br/jspui/handle/20.500.14407/9754-
dc.description.abstractNo presente trabalho, foram utilizados 57 meios para cultivo de Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner 1984, e 26 meios para cultivo de B. garinii Baranton, Postic & Saint Girons 1992. Nestes meios foram adicionadas diferentes substâncias, soros sanguíneos de diferentes espécies animais e vários pHs: 6,4; 6,5; 6,8; 7,0 e 7,5. Observou-se que as duas amostras tiveram um bom crescimento na maioria dos meios testados. Foi estudada a cinética de crescimento de B. burgdorferi e B. garinii, pelo período de 3 meses, utilizando os seguintes meios: 1) BSK adicionado de soro de coelho; 2) BSK adicionado de soro de suino; 3) BSK adicionado de soro de suino e 5 fluorouracil; 4) PMR; 5) CTB; 6) DUBOS; 7) Caldo Brucella e 8) BHI. Todos os meios foram preparados assepticamente e mantidos em tubos de ensaio com capacidade para 10 mL. Para cada meio de cultivo, o inóculo foi padronizado para conter no início, 102 espiroquetas para cada 0,1 mL de cultivo. O monitoramento do crescimento foi feito contando-se o total de espiroquetas em 0,1 mL do meio entre lâmina de microscopia com dimensões de 26 x 76 mm e lamínula com dimensões de 10 x 30 mm, tendo sido utilizado microscópio de campo escuro e aumento de 250X e 400X. A contagem foi realizada pelo período de 14 semanas, tendo sido diária nos primeiros 12 dias e semanal a partir desta data. Houve crescimento de B. burgdorferi e B. garinii, em todos meios testados, com melhor performance para BSK adicionado de soro de coelho, BSK xvi adicionado de soro de suino + 5 fluorouracil e meio CTB. Observou-se crescimento de B. burgdorferi a partir da 4ª semana atingindo o platô de crescimento entre a 8ª e 12ª semanas, quando ocorreu a exaustão do meio de cultivo. Para B. garinii, observou-se um ligeiro aumento do crescimento a partir da 6ª semana, atingindo um platô de crescimento entre a 9ª e 12ª semanas, e seguido da diminuição do mesmo. A B. burgdorferi após ser cultivada em meio de BSK, foi mantida viável em tubo nunc, adicionado de Glicerol a 80%, pelo período de 12 meses a temperatura de 0 ºC. Foi realizada, com sucesso, a coloração das duas amostras de Borrelia, utilizando os corantes cristal violeta, safranina, fucsina e verde brilhante.por
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico - CNPqpor
dc.formatapplication/pdf*
dc.languageporpor
dc.publisherUniversidade Federal Rural do Rio de Janeiropor
dc.rightsAcesso Abertopor
dc.subjectMedicina Veterináriapor
dc.titleCultivo de Borrelia burgdorferi e B. garinii (Spirochaetales:Spirochaetaceae): cinética de crescimento em diferentes meios de cultivo e técnicas de coloraçãopor
dc.typeTesepor
dc.description.abstractOtherIn the present study, it was used 57 media for cultivation of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner 1984, and 26 media for cultivation of Borrelia garinii Baranton, Postic & Saint Girons 1992. In these media it was added different substances, blood serum from different animal species and pH variation of 6.4; 6.5; 6.8, 7.0 and 7.5. It was observed a good growth of the two isolates on most of the tested media. It was studied the dynamic of growth of B. burgdorferi and B. garinii for a period of three months, by utilizing the following media: rabbit serum BSK; 2) swine serum BSK; 3) swine serum BSK + 5- fluorouracil; 4) PMR; 5) CTB; 6) DUBOS; 7) Brucella broth and 8) BHI. All media were prepared aseptically and they were maintained in culture tubes with capacity for 10 mL. For each culture medium, the inoculum was standardized to contain 102 spirochete initially for each 0.1 mL of culture. The monitorization of growth was done by counting the total number of spirochetes in 0.1 mL of medium, utilizing a 26 X 76 mm slide with a 10 x 30 mm cover slip, in a dark field microscope with xviii magnication of 250X or 400X. On the first 12 days counting was done at each 24 hours and after that, once per week, for a period of 14 weeks. There was growth of B. burgdorferi and B. garinii, in all tested medium, with the best performance for BSK with rabbit serum, BSK swine serum + 5 fluorouracil and CTB medium. Growth of B. burgdorferi was observed starting by the 4th week, reaching maximum growth by the 8 th week and 12th week, depleting the culture medium after this time. For B. garinii, it was observed a slight increase in growth at the 6 th week, and a maximum growth between the 9 th and 12 th weeks, and after that, there was reduction of growth. The B. burgdorferi after being cultured in BSK medium, was maintained viable in cryogenic vials with addition of 80% glycerol, for the period of 12 months at 0 ºC. The staining of two isolates of Borrelia was done, with success, by using crystal violet, safranin, fucsin and brilhant green.eng
dc.contributor.advisor1Fonseca, Adivaldo Henrique da
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/4411441162862608por
dc.contributor.referee1Fonseca, Adivaldo Henrique da
dc.contributor.referee2Yoshinari , Natalino Hajime
dc.contributor.referee3Rangel Filho, Francisco Benedito
dc.contributor.referee4Massard, Carlos Luiz
dc.contributor.referee5Inada, Tetsuo
dc.creator.Latteshttp://lattes.cnpq.br/5874811174505909por
dc.publisher.countryBrasilpor
dc.publisher.departmentInstituto de Veterináriapor
dc.publisher.initialsUFRRJpor
dc.publisher.programPrograma de Pós-Graduação em Ciências Veterináriaspor
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dc.thumbnail.urlhttps://tede.ufrrj.br/retrieve/61789/2002%20-%20Angela%20de%20Oliveira.pdf.jpg*
dc.originais.urihttps://tede.ufrrj.br/jspui/handle/jspui/3849
dc.originais.provenanceSubmitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2020-09-09T11:31:17Z No. of bitstreams: 1 2002 - Angela de Oliveira.pdf: 1417105 bytes, checksum: 5aff7eed3c7971bada8de144d1badf25 (MD5)eng
dc.originais.provenanceMade available in DSpace on 2020-09-09T11:31:17Z (GMT). No. of bitstreams: 1 2002 - Angela de Oliveira.pdf: 1417105 bytes, checksum: 5aff7eed3c7971bada8de144d1badf25 (MD5) Previous issue date: 2002-03-21eng
Appears in Collections:Doutorado em Ciências Veterinárias

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