Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa

Detalhes bibliográficos
Autor(a) principal: CAMPOS, Ana Claudia
Data de Publicação: 2012
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRPE
Texto Completo: http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5105
Resumo: Mycoplasma agalactiae is the main causative organism of contagious agalactia (CA), a disease characterized by mastitis followed by agalactia, polyarthritis, and keratoconjunctivitis. In 2001, M. agalactiae was isolated and identified in Brazil, causing great economic losses. Considering the need for characterization of Brazilian strains of M. agalactiae and implementation of effective control, this work was performed in three steps. The first article describes the biochemical and protein profile of isolates of M. agalactiae in small ruminants through the culture in modified Hayflick medium, biochemical tests, polyacrylamide gel electrophoresis (SDS-PAGE) and immunogenicity of these proteins by Western blot (WB). The samples had similar protein in the profile bands ranging from 30-135 kDa in SDS-PAGE, in addition the presence of a protein of 48 kDa immunodominant WB was shown. To continue identifying the agent involved in the AC in Brazil, the second article describes the analysis of ten sequences of M. agalactiae isolated from goats and sheeps. A DNA fragment of 360 bp of the 16S rRNA gene was amplified by PCR and sequenced. The analysis revealed a high degree of similarity among all the sequences. The study revealed greater than 99% similarity with the reference samples of M. agalactiae. In the third article, efficacy of three inactivated vaccines prepared with a local of M. agalactiae sample and adsorbed with three adjuvants was evaluated in goats and sheep. The antibody response in vaccinated animals was analyzed using an indirect ELISA. The three vaccines induced antibody production, and can be an alternative to reduce economic losses and prevent contagious agalactia. These results confirms the presence and spread of M. agalactiae in the country and enhances the possibility of controlling the disease through the adoption of livestock vaccination.
id URPE_d41d79f34360721e606c8a7772979bdd
oai_identifier_str oai:tede2:tede2/5105
network_acronym_str URPE
network_name_str Biblioteca Digital de Teses e Dissertações da UFRPE
repository_id_str
spelling CASTRO, Roberto Soares deAZEVEDO, Edisio Oliveira deNASCIMENTO, Elmiro Rosendo doVESCHI, Josir Laine AparecidaMELO, Marcia Almeida dehttp://lattes.cnpq.br/2920890049034307CAMPOS, Ana Claudia2016-07-28T12:55:08Z2012-02-29CAMPOS, Ana Claudia. Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa. 2012. 75 f.. Tes(Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5105Mycoplasma agalactiae is the main causative organism of contagious agalactia (CA), a disease characterized by mastitis followed by agalactia, polyarthritis, and keratoconjunctivitis. In 2001, M. agalactiae was isolated and identified in Brazil, causing great economic losses. Considering the need for characterization of Brazilian strains of M. agalactiae and implementation of effective control, this work was performed in three steps. The first article describes the biochemical and protein profile of isolates of M. agalactiae in small ruminants through the culture in modified Hayflick medium, biochemical tests, polyacrylamide gel electrophoresis (SDS-PAGE) and immunogenicity of these proteins by Western blot (WB). The samples had similar protein in the profile bands ranging from 30-135 kDa in SDS-PAGE, in addition the presence of a protein of 48 kDa immunodominant WB was shown. To continue identifying the agent involved in the AC in Brazil, the second article describes the analysis of ten sequences of M. agalactiae isolated from goats and sheeps. A DNA fragment of 360 bp of the 16S rRNA gene was amplified by PCR and sequenced. The analysis revealed a high degree of similarity among all the sequences. The study revealed greater than 99% similarity with the reference samples of M. agalactiae. In the third article, efficacy of three inactivated vaccines prepared with a local of M. agalactiae sample and adsorbed with three adjuvants was evaluated in goats and sheep. The antibody response in vaccinated animals was analyzed using an indirect ELISA. The three vaccines induced antibody production, and can be an alternative to reduce economic losses and prevent contagious agalactia. These results confirms the presence and spread of M. agalactiae in the country and enhances the possibility of controlling the disease through the adoption of livestock vaccination.Mycoplasma agalactiae é o principal microrganismo causador da agalaxia contagiosa (AC), doença caracterizada por mastite seguida de agalaxia, poliartrite e ceratoconjuntivite. Em 2001, M. agalactiae foi isolado e identificado no Brasil, determinando grandes prejuízos econômicos. Considerando a necessidade de caracterização de amostras brasileiras de M. agalactiae e da implantação de medidas eficazes de controle, esse trabalho foi realizado em três etapas. O primeiro artigo descreve o perfil bioquímico e protéico de isolados de M. agalactiae de pequenos ruminantes através do cultivo em meio Hayflick modificado, testes bioquímicos e eletroforese em gel de poliacrilamida (SDS-PAGE) e a imunogenicidade destas proteínas por Western Blot (WB). As amostras apresentaram similaridade no perfil protéico com bandas variando de 30 a 135 kDa no SDS-PAGE, além da presença de uma proteína imunodominante de 48 kDa no WB. Para dar continuidade a identificação do agente envolvido na AC no Brasil, o segundo artigo descreve a análise de dez sequências de M. agalactiae isolados de caprinos e ovinos. Um fragmento de DNA de 360 bp do gene 16S rRNA amplificado por PCR foi sequenciado. A análise revelou alto grau de similaridade, entre as dez sequências e uma similaridade maior que 99% com amostras de referência de M. agalactiae. No terceiro artigo, a eficiência de três vacinas inativadas, preparadas com amostra local de M. agalactiae e adsorvidas com três adjuvantes, foi avaliada em caprinos e ovinos. A resposta sorológica dos animais vacinados foi analisada através de um ELISA indireto. As três vacinas induziram produção de anticorpos, podendo ser utilizadas como uma alternativa para reduzir as perdas econômicas e prevenir a agalaxia contagiosa. Estes resultados confirmam a presença e disseminação do M. agalactiae no país e fortalece a possibilidade de controle da doença pela adoção da vacinação dos rebanhos.Submitted by (edna.saturno@ufrpe.br) on 2016-07-28T12:55:08Z No. of bitstreams: 1 Ana Claudia Campos.pdf: 1040308 bytes, checksum: b0653bfe634b8d58d0d6d1c3c04816e9 (MD5)Made available in DSpace on 2016-07-28T12:55:08Z (GMT). No. of bitstreams: 1 Ana Claudia Campos.pdf: 1040308 bytes, checksum: b0653bfe634b8d58d0d6d1c3c04816e9 (MD5) Previous issue date: 2012-02-29Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqapplication/pdfporUniversidade Federal Rural de PernambucoPrograma de Pós-Graduação em Ciência VeterináriaUFRPEBrasilDepartamento de Medicina VeterináriaCaprinoOvinoMicoplasmoseVacinaçãoSequenciamentoWestern BlotGoatSheepMycoplasmosisVaccinationSequencingCIENCIAS AGRARIAS::MEDICINA VETERINARIACaracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-3061482854177903105600600600600-3020210563763616780453670264235017319-2555911436985713659info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFRPEinstname:Universidade Federal Rural de Pernambuco (UFRPE)instacron:UFRPELICENSElicense.txtlicense.txttext/plain; charset=utf-82165http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/5105/1/license.txtbd3efa91386c1718a7f26a329fdcb468MD51ORIGINALAna Claudia Campos.pdfAna Claudia Campos.pdfapplication/pdf1040308http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/5105/2/Ana+Claudia+Campos.pdfb0653bfe634b8d58d0d6d1c3c04816e9MD52tede2/51052016-07-28 09:55:08.247oai:tede2:tede2/5105Tk9UQTogQ09MT1FVRSBBUVVJIEEgU1VBIFBSw5NQUklBIExJQ0VOw4dBCkVzdGEgbGljZW7Dp2EgZGUgZXhlbXBsbyDDqSBmb3JuZWNpZGEgYXBlbmFzIHBhcmEgZmlucyBpbmZvcm1hdGl2b3MuCgpMSUNFTsOHQSBERSBESVNUUklCVUnDh8ODTyBOw4NPLUVYQ0xVU0lWQQoKQ29tIGEgYXByZXNlbnRhw6fDo28gZGVzdGEgbGljZW7Dp2EsIHZvY8OqIChvIGF1dG9yIChlcykgb3UgbyB0aXR1bGFyIGRvcyBkaXJlaXRvcyBkZSBhdXRvcikgY29uY2VkZSDDoCBVbml2ZXJzaWRhZGUgClhYWCAoU2lnbGEgZGEgVW5pdmVyc2lkYWRlKSBvIGRpcmVpdG8gbsOjby1leGNsdXNpdm8gZGUgcmVwcm9kdXppciwgIHRyYWR1emlyIChjb25mb3JtZSBkZWZpbmlkbyBhYmFpeG8pLCBlL291IApkaXN0cmlidWlyIGEgc3VhIHRlc2Ugb3UgZGlzc2VydGHDp8OjbyAoaW5jbHVpbmRvIG8gcmVzdW1vKSBwb3IgdG9kbyBvIG11bmRvIG5vIGZvcm1hdG8gaW1wcmVzc28gZSBlbGV0csO0bmljbyBlIAplbSBxdWFscXVlciBtZWlvLCBpbmNsdWluZG8gb3MgZm9ybWF0b3Mgw6F1ZGlvIG91IHbDrWRlby4KClZvY8OqIGNvbmNvcmRhIHF1ZSBhIFNpZ2xhIGRlIFVuaXZlcnNpZGFkZSBwb2RlLCBzZW0gYWx0ZXJhciBvIGNvbnRlw7pkbywgdHJhbnNwb3IgYSBzdWEgdGVzZSBvdSBkaXNzZXJ0YcOnw6NvIApwYXJhIHF1YWxxdWVyIG1laW8gb3UgZm9ybWF0byBwYXJhIGZpbnMgZGUgcHJlc2VydmHDp8Ojby4KClZvY8OqIHRhbWLDqW0gY29uY29yZGEgcXVlIGEgU2lnbGEgZGUgVW5pdmVyc2lkYWRlIHBvZGUgbWFudGVyIG1haXMgZGUgdW1hIGPDs3BpYSBhIHN1YSB0ZXNlIG91IApkaXNzZXJ0YcOnw6NvIHBhcmEgZmlucyBkZSBzZWd1cmFuw6dhLCBiYWNrLXVwIGUgcHJlc2VydmHDp8Ojby4KClZvY8OqIGRlY2xhcmEgcXVlIGEgc3VhIHRlc2Ugb3UgZGlzc2VydGHDp8OjbyDDqSBvcmlnaW5hbCBlIHF1ZSB2b2PDqiB0ZW0gbyBwb2RlciBkZSBjb25jZWRlciBvcyBkaXJlaXRvcyBjb250aWRvcyAKbmVzdGEgbGljZW7Dp2EuIFZvY8OqIHRhbWLDqW0gZGVjbGFyYSBxdWUgbyBkZXDDs3NpdG8gZGEgc3VhIHRlc2Ugb3UgZGlzc2VydGHDp8OjbyBuw6NvLCBxdWUgc2VqYSBkZSBzZXUgCmNvbmhlY2ltZW50bywgaW5mcmluZ2UgZGlyZWl0b3MgYXV0b3JhaXMgZGUgbmluZ3XDqW0uCgpDYXNvIGEgc3VhIHRlc2Ugb3UgZGlzc2VydGHDp8OjbyBjb250ZW5oYSBtYXRlcmlhbCBxdWUgdm9jw6ogbsOjbyBwb3NzdWkgYSB0aXR1bGFyaWRhZGUgZG9zIGRpcmVpdG9zIGF1dG9yYWlzLCB2b2PDqiAKZGVjbGFyYSBxdWUgb2J0ZXZlIGEgcGVybWlzc8OjbyBpcnJlc3RyaXRhIGRvIGRldGVudG9yIGRvcyBkaXJlaXRvcyBhdXRvcmFpcyBwYXJhIGNvbmNlZGVyIMOgIFNpZ2xhIGRlIFVuaXZlcnNpZGFkZSAKb3MgZGlyZWl0b3MgYXByZXNlbnRhZG9zIG5lc3RhIGxpY2Vuw6dhLCBlIHF1ZSBlc3NlIG1hdGVyaWFsIGRlIHByb3ByaWVkYWRlIGRlIHRlcmNlaXJvcyBlc3TDoSBjbGFyYW1lbnRlIAppZGVudGlmaWNhZG8gZSByZWNvbmhlY2lkbyBubyB0ZXh0byBvdSBubyBjb250ZcO6ZG8gZGEgdGVzZSBvdSBkaXNzZXJ0YcOnw6NvIG9yYSBkZXBvc2l0YWRhLgoKQ0FTTyBBIFRFU0UgT1UgRElTU0VSVEHDh8ODTyBPUkEgREVQT1NJVEFEQSBURU5IQSBTSURPIFJFU1VMVEFETyBERSBVTSBQQVRST0PDjU5JTyBPVSAKQVBPSU8gREUgVU1BIEFHw4pOQ0lBIERFIEZPTUVOVE8gT1UgT1VUUk8gT1JHQU5JU01PIFFVRSBOw4NPIFNFSkEgQSBTSUdMQSBERSAKVU5JVkVSU0lEQURFLCBWT0PDiiBERUNMQVJBIFFVRSBSRVNQRUlUT1UgVE9ET1MgRSBRVUFJU1FVRVIgRElSRUlUT1MgREUgUkVWSVPDg08gQ09NTyAKVEFNQsOJTSBBUyBERU1BSVMgT0JSSUdBw4fDlUVTIEVYSUdJREFTIFBPUiBDT05UUkFUTyBPVSBBQ09SRE8uCgpBIFNpZ2xhIGRlIFVuaXZlcnNpZGFkZSBzZSBjb21wcm9tZXRlIGEgaWRlbnRpZmljYXIgY2xhcmFtZW50ZSBvIHNldSBub21lIChzKSBvdSBvKHMpIG5vbWUocykgZG8ocykgCmRldGVudG9yKGVzKSBkb3MgZGlyZWl0b3MgYXV0b3JhaXMgZGEgdGVzZSBvdSBkaXNzZXJ0YcOnw6NvLCBlIG7Do28gZmFyw6EgcXVhbHF1ZXIgYWx0ZXJhw6fDo28sIGFsw6ltIGRhcXVlbGFzIApjb25jZWRpZGFzIHBvciBlc3RhIGxpY2Vuw6dhLgo=Biblioteca Digital de Teses e Dissertaçõeshttp://www.tede2.ufrpe.br:8080/tede/PUBhttp://www.tede2.ufrpe.br:8080/oai/requestbdtd@ufrpe.br ||bdtd@ufrpe.bropendoar:2016-07-28T12:55:08Biblioteca Digital de Teses e Dissertações da UFRPE - Universidade Federal Rural de Pernambuco (UFRPE)false
dc.title.por.fl_str_mv Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
title Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
spellingShingle Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
CAMPOS, Ana Claudia
Caprino
Ovino
Micoplasmose
Vacinação
Sequenciamento
Western Blot
Goat
Sheep
Mycoplasmosis
Vaccination
Sequencing
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
title_full Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
title_fullStr Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
title_full_unstemmed Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
title_sort Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa
author CAMPOS, Ana Claudia
author_facet CAMPOS, Ana Claudia
author_role author
dc.contributor.advisor1.fl_str_mv CASTRO, Roberto Soares de
dc.contributor.advisor-co1.fl_str_mv AZEVEDO, Edisio Oliveira de
dc.contributor.referee1.fl_str_mv NASCIMENTO, Elmiro Rosendo do
dc.contributor.referee2.fl_str_mv VESCHI, Josir Laine Aparecida
dc.contributor.referee3.fl_str_mv MELO, Marcia Almeida de
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2920890049034307
dc.contributor.author.fl_str_mv CAMPOS, Ana Claudia
contributor_str_mv CASTRO, Roberto Soares de
AZEVEDO, Edisio Oliveira de
NASCIMENTO, Elmiro Rosendo do
VESCHI, Josir Laine Aparecida
MELO, Marcia Almeida de
dc.subject.por.fl_str_mv Caprino
Ovino
Micoplasmose
Vacinação
Sequenciamento
topic Caprino
Ovino
Micoplasmose
Vacinação
Sequenciamento
Western Blot
Goat
Sheep
Mycoplasmosis
Vaccination
Sequencing
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.eng.fl_str_mv Western Blot
Goat
Sheep
Mycoplasmosis
Vaccination
Sequencing
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Mycoplasma agalactiae is the main causative organism of contagious agalactia (CA), a disease characterized by mastitis followed by agalactia, polyarthritis, and keratoconjunctivitis. In 2001, M. agalactiae was isolated and identified in Brazil, causing great economic losses. Considering the need for characterization of Brazilian strains of M. agalactiae and implementation of effective control, this work was performed in three steps. The first article describes the biochemical and protein profile of isolates of M. agalactiae in small ruminants through the culture in modified Hayflick medium, biochemical tests, polyacrylamide gel electrophoresis (SDS-PAGE) and immunogenicity of these proteins by Western blot (WB). The samples had similar protein in the profile bands ranging from 30-135 kDa in SDS-PAGE, in addition the presence of a protein of 48 kDa immunodominant WB was shown. To continue identifying the agent involved in the AC in Brazil, the second article describes the analysis of ten sequences of M. agalactiae isolated from goats and sheeps. A DNA fragment of 360 bp of the 16S rRNA gene was amplified by PCR and sequenced. The analysis revealed a high degree of similarity among all the sequences. The study revealed greater than 99% similarity with the reference samples of M. agalactiae. In the third article, efficacy of three inactivated vaccines prepared with a local of M. agalactiae sample and adsorbed with three adjuvants was evaluated in goats and sheep. The antibody response in vaccinated animals was analyzed using an indirect ELISA. The three vaccines induced antibody production, and can be an alternative to reduce economic losses and prevent contagious agalactia. These results confirms the presence and spread of M. agalactiae in the country and enhances the possibility of controlling the disease through the adoption of livestock vaccination.
publishDate 2012
dc.date.issued.fl_str_mv 2012-02-29
dc.date.accessioned.fl_str_mv 2016-07-28T12:55:08Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv CAMPOS, Ana Claudia. Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa. 2012. 75 f.. Tes(Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.
dc.identifier.uri.fl_str_mv http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5105
identifier_str_mv CAMPOS, Ana Claudia. Caracterização de Mycoplasma agalactiae isolados no Brasil e produção de vacinas contra agalaxia contagiosa. 2012. 75 f.. Tes(Programa de Pós-Graduação em Ciência Veterinária) - Universidade Federal Rural de Pernambuco, Recife.
url http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5105
dc.language.iso.fl_str_mv por
language por
dc.relation.program.fl_str_mv -3061482854177903105
dc.relation.confidence.fl_str_mv 600
600
600
600
dc.relation.department.fl_str_mv -3020210563763616780
dc.relation.cnpq.fl_str_mv 453670264235017319
dc.relation.sponsorship.fl_str_mv -2555911436985713659
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal Rural de Pernambuco
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Ciência Veterinária
dc.publisher.initials.fl_str_mv UFRPE
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Departamento de Medicina Veterinária
publisher.none.fl_str_mv Universidade Federal Rural de Pernambuco
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações da UFRPE
instname:Universidade Federal Rural de Pernambuco (UFRPE)
instacron:UFRPE
instname_str Universidade Federal Rural de Pernambuco (UFRPE)
instacron_str UFRPE
institution UFRPE
reponame_str Biblioteca Digital de Teses e Dissertações da UFRPE
collection Biblioteca Digital de Teses e Dissertações da UFRPE
bitstream.url.fl_str_mv http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/5105/1/license.txt
http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/5105/2/Ana+Claudia+Campos.pdf
bitstream.checksum.fl_str_mv bd3efa91386c1718a7f26a329fdcb468
b0653bfe634b8d58d0d6d1c3c04816e9
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da UFRPE - Universidade Federal Rural de Pernambuco (UFRPE)
repository.mail.fl_str_mv bdtd@ufrpe.br ||bdtd@ufrpe.br
_version_ 1800311450180255744