Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas

Detalhes bibliográficos
Ano de defesa: 2016
Autor(a) principal: Natália Rocha Guimarães
Orientador(a): Não Informado pela instituição
Banca de defesa: Não Informado pela instituição
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Minas Gerais
Brasil
ICB - DEPARTAMENTO DE MICROBIOLOGIA
Programa de Pós-Graduação em Microbiologia
UFMG
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://hdl.handle.net/1843/42406
https://orcid.org/0000-0002-9859-5895
Resumo: The search for new antimicrobial substances, especially naturally occurring ones, are gaining great prominence, being the target of several studies, in particular due to the emergence of multiresistant microorganisms, as well as their association with the occurrence of outbreaks. Acinetobacter baumannii is a bacterium of great clinical relevance, that markedly shows multidrug resistance. The aim of this study was to evaluate and to characterize the antagonistic activity of A. baumanni. Among the 19 tested strains, only one, A. baumannii 397, expressed antagonism. Activity was observed exclusively against isolates of the same species. The interference of bacteriophages, fatty acids, residual chloroform and hydrogen peroxide in the interpretation of data was evaluated. In order to optimize antagonism expression, the producer strain was cultivated under different conditions. When grown at 25 °C, larger and clear inhibition zones were generated. Protein extraction of A. baumannii 397 was performed by acid precipitation with HCl 5N. The antagonistic activity was confirmed by the spread plate technique against a clinical isolate of A. baumannii. The crude extract showed antagonist activity of 1280 AU/mL. The extract was inactivated by proteases and remained active after exposure to organic solvents and a broad pH range and temperatures. CIM and CBM values showed bactericidal activity against the indicator strain. The crude extract was submitted to three steps of reverse-phase chromatography, the first step in a C-8 column and the others in a C-18 column. The last chromatographic step generated two active fractions, C5 and C6, eluted with approximately 80% acetonitrile. These fractions were submitted to MS. Five molecular masses were detected in C5, and in C6, two molecular masses of 6.2 kDa and 8.3 kDa. The results showed the synthesis of hydrophobic antimicrobial proteinaceous substance(s), active(s) against multidrug-resistant bacteria, by A. baumannii 397, property not yet described for this species, that displays biotechnological potential.
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spelling Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianasAcinetobacter baumanniiSubstância antagonistaBacteriocinaMicrobiologiaAcinetobacter baumanniiBacteriocinasThe search for new antimicrobial substances, especially naturally occurring ones, are gaining great prominence, being the target of several studies, in particular due to the emergence of multiresistant microorganisms, as well as their association with the occurrence of outbreaks. Acinetobacter baumannii is a bacterium of great clinical relevance, that markedly shows multidrug resistance. The aim of this study was to evaluate and to characterize the antagonistic activity of A. baumanni. Among the 19 tested strains, only one, A. baumannii 397, expressed antagonism. Activity was observed exclusively against isolates of the same species. The interference of bacteriophages, fatty acids, residual chloroform and hydrogen peroxide in the interpretation of data was evaluated. In order to optimize antagonism expression, the producer strain was cultivated under different conditions. When grown at 25 °C, larger and clear inhibition zones were generated. Protein extraction of A. baumannii 397 was performed by acid precipitation with HCl 5N. The antagonistic activity was confirmed by the spread plate technique against a clinical isolate of A. baumannii. The crude extract showed antagonist activity of 1280 AU/mL. The extract was inactivated by proteases and remained active after exposure to organic solvents and a broad pH range and temperatures. CIM and CBM values showed bactericidal activity against the indicator strain. The crude extract was submitted to three steps of reverse-phase chromatography, the first step in a C-8 column and the others in a C-18 column. The last chromatographic step generated two active fractions, C5 and C6, eluted with approximately 80% acetonitrile. These fractions were submitted to MS. Five molecular masses were detected in C5, and in C6, two molecular masses of 6.2 kDa and 8.3 kDa. The results showed the synthesis of hydrophobic antimicrobial proteinaceous substance(s), active(s) against multidrug-resistant bacteria, by A. baumannii 397, property not yet described for this species, that displays biotechnological potential.A busca por novas substâncias antimicrobianas, principalmente de ocorrência natural, vêm ganhando grande destaque, sendo alvo de diversos estudos, em especial, devido à emergência de microrganismos resistentes, bem como à sua associação com a ocorrência de surtos. Acinetobacter baumannii é uma bactéria de grande relevância clínica, marcadamente multirresistente a antimicrobianos. Este estudo foi realizado com o objetivo de avaliar e caracterizar a atividade antagonista de A. baumannii. Dentre as 19 amostras testadas, somente uma, A. baumannii 397, expressou antagonismo. Detectou-se atividade apenas contra amostras da mesma espécie. Avaliou-se a possibilidade de interferência de bacteriófagos, ácidos graxos, clorofórmio residual e peróxido de hidrogênio na interpretação dos dados. Visando otimizar a expressão do antagonismo, a amostra produtora foi cultivada sob diferentes condições. Quando cultivada à temperatura de 25 ºC, os halos de inibição eram maiores e mais límpidos. A extração proteica da amostra A. baumannii 397 foi realizada por meio de precipitação ácida com HCl 5 N. A atividade antagonista foi confirmada por técnica de spread plate contra amostra clínica de A. baumannii. O extrato bruto apresentou atividade antagonista de 1280 UA/mL. O extrato foi inativado por proteases e manteve-se ativo após exposição a solventes orgânicos e uma ampla faixa de pH e temperaturas. Os valores de CIM e CBM mostraram atividade bactericida contra a amostra reveladora. O extrato bruto foi submetido a três etapas de cromatografia de fase reversa, sendo, a primeira etapa em coluna C-8 e, as demais, em coluna C-18. A última etapa cromatográfica gerou duas frações ativas, C5 e C6, eluídas com, aproximadamente, 80% de acetonitrila. Essas frações foram submetidas à MS. Cinco massas moleculares foram detectadas em C5 e, em C6, duas massas moleculares, de 6,2 kDa e 8,3 kDa. Os dados mostram a síntese de substância(s) antagonista(s) hidrofóbica(s), de natureza proteica, ativa(s) contra bactérias multirresistentes, pela amostra A. baumannii 397, propriedade ainda não descrita para a espécie, com potencial para aplicação biotecnológica.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorOutra AgênciaUniversidade Federal de Minas GeraisBrasilICB - DEPARTAMENTO DE MICROBIOLOGIAPrograma de Pós-Graduação em MicrobiologiaUFMGPaula Prazeres Magalhãeshttp://lattes.cnpq.br/2184773662680617Luiz de Macêdo FariasEdmar Chartone de SouzaAna Carolina Morais ApolônioSílvia Beleza de MouraNatália Rocha Guimarães2022-06-09T19:03:03Z2022-06-09T19:03:03Z2016-01-18info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/1843/42406https://orcid.org/0000-0002-9859-5895porhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2022-06-09T19:03:04Zoai:repositorio.ufmg.br:1843/42406Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2022-06-09T19:03:04Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
title Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
spellingShingle Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
Natália Rocha Guimarães
Acinetobacter baumannii
Substância antagonista
Bacteriocina
Microbiologia
Acinetobacter baumannii
Bacteriocinas
title_short Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
title_full Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
title_fullStr Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
title_full_unstemmed Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
title_sort Atividade antagonista de Acinetobacter baumannii contra amostras de Acinetobacter resistentes a drogas antimicrobianas
author Natália Rocha Guimarães
author_facet Natália Rocha Guimarães
author_role author
dc.contributor.none.fl_str_mv Paula Prazeres Magalhães
http://lattes.cnpq.br/2184773662680617
Luiz de Macêdo Farias
Edmar Chartone de Souza
Ana Carolina Morais Apolônio
Sílvia Beleza de Moura
dc.contributor.author.fl_str_mv Natália Rocha Guimarães
dc.subject.por.fl_str_mv Acinetobacter baumannii
Substância antagonista
Bacteriocina
Microbiologia
Acinetobacter baumannii
Bacteriocinas
topic Acinetobacter baumannii
Substância antagonista
Bacteriocina
Microbiologia
Acinetobacter baumannii
Bacteriocinas
description The search for new antimicrobial substances, especially naturally occurring ones, are gaining great prominence, being the target of several studies, in particular due to the emergence of multiresistant microorganisms, as well as their association with the occurrence of outbreaks. Acinetobacter baumannii is a bacterium of great clinical relevance, that markedly shows multidrug resistance. The aim of this study was to evaluate and to characterize the antagonistic activity of A. baumanni. Among the 19 tested strains, only one, A. baumannii 397, expressed antagonism. Activity was observed exclusively against isolates of the same species. The interference of bacteriophages, fatty acids, residual chloroform and hydrogen peroxide in the interpretation of data was evaluated. In order to optimize antagonism expression, the producer strain was cultivated under different conditions. When grown at 25 °C, larger and clear inhibition zones were generated. Protein extraction of A. baumannii 397 was performed by acid precipitation with HCl 5N. The antagonistic activity was confirmed by the spread plate technique against a clinical isolate of A. baumannii. The crude extract showed antagonist activity of 1280 AU/mL. The extract was inactivated by proteases and remained active after exposure to organic solvents and a broad pH range and temperatures. CIM and CBM values showed bactericidal activity against the indicator strain. The crude extract was submitted to three steps of reverse-phase chromatography, the first step in a C-8 column and the others in a C-18 column. The last chromatographic step generated two active fractions, C5 and C6, eluted with approximately 80% acetonitrile. These fractions were submitted to MS. Five molecular masses were detected in C5, and in C6, two molecular masses of 6.2 kDa and 8.3 kDa. The results showed the synthesis of hydrophobic antimicrobial proteinaceous substance(s), active(s) against multidrug-resistant bacteria, by A. baumannii 397, property not yet described for this species, that displays biotechnological potential.
publishDate 2016
dc.date.none.fl_str_mv 2016-01-18
2022-06-09T19:03:03Z
2022-06-09T19:03:03Z
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https://orcid.org/0000-0002-9859-5895
url http://hdl.handle.net/1843/42406
https://orcid.org/0000-0002-9859-5895
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dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
Brasil
ICB - DEPARTAMENTO DE MICROBIOLOGIA
Programa de Pós-Graduação em Microbiologia
UFMG
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
Brasil
ICB - DEPARTAMENTO DE MICROBIOLOGIA
Programa de Pós-Graduação em Microbiologia
UFMG
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
instacron_str UFMG
institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv repositorio@ufmg.br
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