Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães

Detalhes bibliográficos
Ano de defesa: 2012
Autor(a) principal: Pieri, Fábio Alessandro
Orientador(a): Moreira, Maria Aparecida Scatamburlo lattes
Banca de defesa: Gioso, Marco Antônio lattes, Mantovani, Hilário Cuquetto lattes
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Viçosa
Programa de Pós-Graduação: Doutorado em Medicina Veterinária
Departamento: Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://locus.ufv.br/handle/123456789/1467
Resumo: Periodontal disease affects about 85% of dogs older than four years and is caused by the buildup of plaque on the tooth surface and its body's reaction.In other work, Copaiba oil presented positive results in the inhibition of dental plaque in dogs, however the literature do not present any study that identifying this activity against bacteria from dog s dental plaque "in vitro" and to establishing the oil active substances. The objective of this study was to evaluate the antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluate the bioproduct obtained in the inhibition of dental plaque bacteria in dogs. C. langsdorffii oil was used, subjected to bioassay-guided fractionation and its last fraction had its compounds identified. The supragingival dental plaque of ten dogs aged between one and two years was obtained, grown on Mitis Salivarius agar with serial dilutions to obtain isolated colonies that was cultivated to pure cultures. The bacteria were identified using the PCR technique to amplify a part of the 16S rDNA gene and the amplified fragments were sequenced and compared with the sequences deposited in GenBank. The oil s antimicrobial activity was tested by agar diffusion test against 27 isolates of Escherichia coli from mastitic milk, an field isolated of Actinobacillus pleuropneumoniae and Haemophilus parasuis and reference strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Shigella flexneri and Citrobacter freundi. The antibacterial ability of oil and its fractions was tested against 20 isolates of dog s dental plaque bacteria with the same methodology. One of the major compounds identified in the last fraction obtained was selected and had its minimum inhibitory concentration evaluated against 34 isolates of dental plaque bacteria from dogs and was tested in an assay using dogs to check their ability to inhibit plaque formation "in vivo" compared to a negative control and a positive with 0.125% chlorhexidine. Dental plaque was obtained from 10 dogs and 183 isolates were isolated and identified with a predominance of the genera Staphylococcus (20.2%), Enterococcus (19.1%) and Streptococcus (18%). Copaiba oil inhibited the standard strains of E. coli, S. flexneri, P. aeruginosa and S. aureus with inhibition zones with 17, 18, 15 and 18mm respectively. Eight wild strains of E. coli were sensitive to the oil showing halos ranging among 12 to 34mm. When tested on dental plaque bacteria isolated from dogs, copaiba oil inhibited 16/20 isolates with halos ranging among 10 and 15mm, and with respect to its constituints, the not acid fraction was more effective than acid fraction, with activity against 17/20 and 8/20 isolates respectively. The last fraction was obtained from the not acid and was able to inhibit 18/20 of target isolates. The selected compound was called LDBACUFV01 and had established its minimum inhibitory concentration against the isolates, being able to inhibit 72% and was considered qualified for the "in vivo" assay. The bioproduct was applied in dogs and had better effect in reducing dental plaque the formation than negative control (p<0.001) and positive control (p<0.05), with plaque coverage about 23.3%, 65% and 37.5% of total area of teeth respectively. In conclusion, copaiba oil presented antibacterial activity against bacteria of dogs dental plaque and a bioproduct obtained, with a sesquiterpene component of this oil, may be used in reduction of dental plaque in dogs.
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spelling Pieri, Fábio Alessandrohttp://lattes.cnpq.br/1622593528670181Silva Júnior, Abelardohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4771645P8Borges, Andrea Pacheco Batistahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767373D8Moreira, Maria Aparecida Scatamburlohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797678J6Gioso, Marco Antôniohttp://lattes.cnpq.br/5242351363604381Mantovani, Hilário Cuquettohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026Z72015-03-26T12:47:50Z2014-06-092015-03-26T12:47:50Z2012-02-16PIERI, Fábio Alessandro. Antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluation of obtained bioproduct in inhibition of dental plaque bacteria in dogs. 2012. 112 f. Tese (Doutorado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.http://locus.ufv.br/handle/123456789/1467Periodontal disease affects about 85% of dogs older than four years and is caused by the buildup of plaque on the tooth surface and its body's reaction.In other work, Copaiba oil presented positive results in the inhibition of dental plaque in dogs, however the literature do not present any study that identifying this activity against bacteria from dog s dental plaque "in vitro" and to establishing the oil active substances. The objective of this study was to evaluate the antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluate the bioproduct obtained in the inhibition of dental plaque bacteria in dogs. C. langsdorffii oil was used, subjected to bioassay-guided fractionation and its last fraction had its compounds identified. The supragingival dental plaque of ten dogs aged between one and two years was obtained, grown on Mitis Salivarius agar with serial dilutions to obtain isolated colonies that was cultivated to pure cultures. The bacteria were identified using the PCR technique to amplify a part of the 16S rDNA gene and the amplified fragments were sequenced and compared with the sequences deposited in GenBank. The oil s antimicrobial activity was tested by agar diffusion test against 27 isolates of Escherichia coli from mastitic milk, an field isolated of Actinobacillus pleuropneumoniae and Haemophilus parasuis and reference strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Shigella flexneri and Citrobacter freundi. The antibacterial ability of oil and its fractions was tested against 20 isolates of dog s dental plaque bacteria with the same methodology. One of the major compounds identified in the last fraction obtained was selected and had its minimum inhibitory concentration evaluated against 34 isolates of dental plaque bacteria from dogs and was tested in an assay using dogs to check their ability to inhibit plaque formation "in vivo" compared to a negative control and a positive with 0.125% chlorhexidine. Dental plaque was obtained from 10 dogs and 183 isolates were isolated and identified with a predominance of the genera Staphylococcus (20.2%), Enterococcus (19.1%) and Streptococcus (18%). Copaiba oil inhibited the standard strains of E. coli, S. flexneri, P. aeruginosa and S. aureus with inhibition zones with 17, 18, 15 and 18mm respectively. Eight wild strains of E. coli were sensitive to the oil showing halos ranging among 12 to 34mm. When tested on dental plaque bacteria isolated from dogs, copaiba oil inhibited 16/20 isolates with halos ranging among 10 and 15mm, and with respect to its constituints, the not acid fraction was more effective than acid fraction, with activity against 17/20 and 8/20 isolates respectively. The last fraction was obtained from the not acid and was able to inhibit 18/20 of target isolates. The selected compound was called LDBACUFV01 and had established its minimum inhibitory concentration against the isolates, being able to inhibit 72% and was considered qualified for the "in vivo" assay. The bioproduct was applied in dogs and had better effect in reducing dental plaque the formation than negative control (p<0.001) and positive control (p<0.05), with plaque coverage about 23.3%, 65% and 37.5% of total area of teeth respectively. In conclusion, copaiba oil presented antibacterial activity against bacteria of dogs dental plaque and a bioproduct obtained, with a sesquiterpene component of this oil, may be used in reduction of dental plaque in dogs.A doença periodontal acomete cerca de 85% dos cães com mais de quatro anos de idade e é causada pelo acúmulo de placa bacteriana na superfície dos dentes e pela reação do organismo a esta. O óleo de copaíba apresentou, anteriormente, resultados positivos na inibição da placa dental em cães, entretanto faltam estudos que identifiquem esta atividade sobre as bactérias oriundas de cães in vitro e que estabeleçam as substâncias ativas do óleo. O objetivo deste trabalho foi verificar a atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliar o bioproduto obtido na inibição de bactérias da placa dental de cães. Foi utilizado um óleo de C. langsdorffii, submetido a fracionamento guiado por bioensaio e sua última fração teve seus compostos identificados. Para confecção da bacterioteca deste trabalho a placa dental supragengival de dez cães com idade entre um e dois anos foi obtida, cultivada em ágar Mitis Salivarius com diluições seriadas para obterem-se colônias isoladas e culturas puras a partir delas. As bactérias foram identificadas utilizando a técnica de PCR para amplificação de parte do gene 16S do rDNA e os fragmentos amplificados foram sequenciados e comparados com as seqüências depositadas no GenBank. O óleo íntegro teve sua capacidade antimicrobiana testada, por teste de difusão em ágar, contra 27 isolados de Escherichia coli provenientes de leite mastítico, um isolado de campo de Actinobacilus pleuropneumoniae e de Haemophilus parasuis além de cepas de referência de Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Shigella flexneri e Citrobacter freundi. A capacidade antibacteriana do óleo íntegro e de suas frações iniciais foi testada contra 20 isolados da bacterioteca da placa dental com a mesma metodologia. Dos compostos majoritários identificados na última fração obtida foi selecionado um que teve sua concentração inibitória mínima avaliada sobre 34 isolados da bacterioteca e testado em ensaio com cães para verificar sua capacidade de inibir a formação de placa in vivo comparado a um controle negativo e um positivo com clorexidina 0,125%. Da placa dental dos 10 cães foi obtida uma bacterioteca de 183 isolados com predominância dos gêneros Staphylococcus (20,2%), Enterococcus (19,1%) e Streptococcus(18%). O óleo integro inibiu o crescimento das cepas padrão de E. coli, S. flexneri, P. aeruginosa e S. aureus com halos de inibição de 17, 18, 15 e 18mm respectivamente. Oito isolados selvagens de E. coli foram sensíveis ao óleo apresentando halos que variaram de 12 a 34mm. Quando testado sobre isolados da placa dental de cães, o óleo de copaíba inibiu 16/20 isolados de diversos gêneros com halos variando entre 10 e 15mm, e com relação às suas frações, a não ácida se apresentou superior, com atividade sobre 17/20 isolados e foi refracionada, por ter ação contra mais isolados do que a fração ácida, que inibiu apenas 8/20 isolados. A última fração foi obtida da não ácida e foi capaz de inibir 18/20 isolados da bacterioteca da placa dental de cães. O composto selecionado foi denominado LDBACUFV01 e teve sua concentração inibitória mínima estabelecida sobre 33 isolados da bacterioteca, sendo capaz de inibir 72% e foi considerado habilitado para o ensaio in vivo . Um bioproduto foi formulado com este composto e aplicado nos dentes de cães por 15 dias, sendo considerado superior ao controle negativo (p<0,001) e positivo (p<0,05) na redução da formação de placa bacteriana na superfície dental, pois a área de cobertura de placa nos dentes dos cães foi de 23,3%, 65% e 37,5% respectivamente. Conclui-se que o óleo de copaíba possui capacidade antimicrobiana sobre bactérias componentes da placa dental de cães e que o bioproduto obtido, com um composto da fração de hidrocarbonetos sesquiterpênicos do óleo, pode ser utilizado para redução da placa dental de cães.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaDoutorado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deFitoterápicosCopaibaPeriodontiaCãesHerbal MedicinesCopaibaPeriodonticsdogsCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::DOENCAS INFECCIOSAS DE ANIMAISAtividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cãesAntimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluation of obtained bioproduct in inhibition of dental plaque bacteria in dogsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf2055846https://locus.ufv.br//bitstream/123456789/1467/1/texto%20completo.pdf329a4b063f0860451bb387d7911a6ff4MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain177321https://locus.ufv.br//bitstream/123456789/1467/2/texto%20completo.pdf.txt8218d200d0b94ec776c57d06c1370c2eMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3776https://locus.ufv.br//bitstream/123456789/1467/3/texto%20completo.pdf.jpg05423ed38ffa27ce7983d754a724bc11MD53123456789/14672016-04-07 23:04:16.469oai:locus.ufv.br:123456789/1467Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-08T02:04:16LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
dc.title.alternative.eng.fl_str_mv Antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluation of obtained bioproduct in inhibition of dental plaque bacteria in dogs
title Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
spellingShingle Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
Pieri, Fábio Alessandro
Fitoterápicos
Copaiba
Periodontia
Cães
Herbal Medicines
Copaiba
Periodontics
dogs
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::DOENCAS INFECCIOSAS DE ANIMAIS
title_short Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
title_full Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
title_fullStr Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
title_full_unstemmed Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
title_sort Atividade antimicrobiana do óleo de copaíba (Copaifera langsdorffii) e seus constituintes, e avaliação do bioproduto obtido na inibição de bactérias da placa dental de cães
author Pieri, Fábio Alessandro
author_facet Pieri, Fábio Alessandro
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/1622593528670181
dc.contributor.author.fl_str_mv Pieri, Fábio Alessandro
dc.contributor.advisor-co1.fl_str_mv Silva Júnior, Abelardo
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4771645P8
dc.contributor.advisor-co2.fl_str_mv Borges, Andrea Pacheco Batista
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767373D8
dc.contributor.advisor1.fl_str_mv Moreira, Maria Aparecida Scatamburlo
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797678J6
dc.contributor.referee1.fl_str_mv Gioso, Marco Antônio
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/5242351363604381
dc.contributor.referee2.fl_str_mv Mantovani, Hilário Cuquetto
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026Z7
contributor_str_mv Silva Júnior, Abelardo
Borges, Andrea Pacheco Batista
Moreira, Maria Aparecida Scatamburlo
Gioso, Marco Antônio
Mantovani, Hilário Cuquetto
dc.subject.por.fl_str_mv Fitoterápicos
Copaiba
Periodontia
Cães
topic Fitoterápicos
Copaiba
Periodontia
Cães
Herbal Medicines
Copaiba
Periodontics
dogs
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::DOENCAS INFECCIOSAS DE ANIMAIS
dc.subject.eng.fl_str_mv Herbal Medicines
Copaiba
Periodontics
dogs
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::DOENCAS INFECCIOSAS DE ANIMAIS
description Periodontal disease affects about 85% of dogs older than four years and is caused by the buildup of plaque on the tooth surface and its body's reaction.In other work, Copaiba oil presented positive results in the inhibition of dental plaque in dogs, however the literature do not present any study that identifying this activity against bacteria from dog s dental plaque "in vitro" and to establishing the oil active substances. The objective of this study was to evaluate the antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluate the bioproduct obtained in the inhibition of dental plaque bacteria in dogs. C. langsdorffii oil was used, subjected to bioassay-guided fractionation and its last fraction had its compounds identified. The supragingival dental plaque of ten dogs aged between one and two years was obtained, grown on Mitis Salivarius agar with serial dilutions to obtain isolated colonies that was cultivated to pure cultures. The bacteria were identified using the PCR technique to amplify a part of the 16S rDNA gene and the amplified fragments were sequenced and compared with the sequences deposited in GenBank. The oil s antimicrobial activity was tested by agar diffusion test against 27 isolates of Escherichia coli from mastitic milk, an field isolated of Actinobacillus pleuropneumoniae and Haemophilus parasuis and reference strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Shigella flexneri and Citrobacter freundi. The antibacterial ability of oil and its fractions was tested against 20 isolates of dog s dental plaque bacteria with the same methodology. One of the major compounds identified in the last fraction obtained was selected and had its minimum inhibitory concentration evaluated against 34 isolates of dental plaque bacteria from dogs and was tested in an assay using dogs to check their ability to inhibit plaque formation "in vivo" compared to a negative control and a positive with 0.125% chlorhexidine. Dental plaque was obtained from 10 dogs and 183 isolates were isolated and identified with a predominance of the genera Staphylococcus (20.2%), Enterococcus (19.1%) and Streptococcus (18%). Copaiba oil inhibited the standard strains of E. coli, S. flexneri, P. aeruginosa and S. aureus with inhibition zones with 17, 18, 15 and 18mm respectively. Eight wild strains of E. coli were sensitive to the oil showing halos ranging among 12 to 34mm. When tested on dental plaque bacteria isolated from dogs, copaiba oil inhibited 16/20 isolates with halos ranging among 10 and 15mm, and with respect to its constituints, the not acid fraction was more effective than acid fraction, with activity against 17/20 and 8/20 isolates respectively. The last fraction was obtained from the not acid and was able to inhibit 18/20 of target isolates. The selected compound was called LDBACUFV01 and had established its minimum inhibitory concentration against the isolates, being able to inhibit 72% and was considered qualified for the "in vivo" assay. The bioproduct was applied in dogs and had better effect in reducing dental plaque the formation than negative control (p<0.001) and positive control (p<0.05), with plaque coverage about 23.3%, 65% and 37.5% of total area of teeth respectively. In conclusion, copaiba oil presented antibacterial activity against bacteria of dogs dental plaque and a bioproduct obtained, with a sesquiterpene component of this oil, may be used in reduction of dental plaque in dogs.
publishDate 2012
dc.date.issued.fl_str_mv 2012-02-16
dc.date.available.fl_str_mv 2014-06-09
2015-03-26T12:47:50Z
dc.date.accessioned.fl_str_mv 2015-03-26T12:47:50Z
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dc.identifier.citation.fl_str_mv PIERI, Fábio Alessandro. Antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluation of obtained bioproduct in inhibition of dental plaque bacteria in dogs. 2012. 112 f. Tese (Doutorado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/1467
identifier_str_mv PIERI, Fábio Alessandro. Antimicrobial activity of copaiba oil (Copaifera langsdorffii) and its constituents, and evaluation of obtained bioproduct in inhibition of dental plaque bacteria in dogs. 2012. 112 f. Tese (Doutorado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
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