Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2014 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/490 |
Resumo: | The high occurrence of Campylobacter spp. in chicken meat and by- products and their increasing antimicrobial resistance have worried public health authorities worldwide. The objectives of this study were to evaluate the Campylobacter contamination in chicken carcasses and antimicrobial resistance of the isolates, as well as to investigate the resistance-modifying and efflux inhibitory activities of plant extract against this microorganism. A total of 95 chicken carcasses from slaughterhouses from Minas Gerais Brazil were analyzed by most probable number method (MPN) associated with subsequent confirmation by polymerase chain reaction (PCR). The isolates were screened for antimicrobial resistance by agar diffusion disk method, and the main genes and mechanisms of resistance were investigated. Twenty ethanolic extracts of plants were tested for antimicrobial, resistance-modifying and accumulation of ethidium bromide activities. The major classes of secondary metabolites in the plant extracts showing biological activities were analyzed by high performance liquid chromatography (HPLC). Campylobacter spp. was found in 16.8% of samples with microbial load ranging from 60.0 to 184 MPN/carcass, and C. jejuni was the species most prevalent (93.8 %). The isolates were resistant to at least 5 (31.2 %) of the antimicrobial agents tested, and high resistance rates were observed for cephalosporins (75 - 100 %), quinolones (95 %), tetracycline (50 %) and ampicillin (45 %). Ciprofloxacin resistance proved to be caused by mutation in the gyrA gene, whereas 40 % of the tetracycline resistance was explained by presence of the tet(O) gene. The blaOXA-61 gene, which encodes for a β-lactamase, was found in both resistant and susceptible isolates for ampicillin. Almost all isolates showed the three genes required to synthesize the CmeABC efflux system. The use of phenylalanine-arginine β-naphthylamide (PAβN), a well-known efflux pump inhibitor, decreased the MIC of ciprofloxacin, tetracycline, ampicillin and bile salts by 2 to 8 fold confirming the activity of this resistance mechanism. These extracts showed weak antimicrobial activity comparing to antibiotics, with MIC values ranging from 0.1 to >16.38 mg/mL. However, most of them showed synergistic effects on combining with bile salts when used at subinhibitory concentration, decreasing 2 to 16 fold the MIC values of these antimicrobials. We observed that ten plant extracts increased the accumulation of ethidium bromide in the Campylobacter cells, suggesting inhibition of efflux systems. The extracts of Alpinia zerumbet and Vernonia polyanthes showed greater synergistic effect on combining with bile salts, whereas extracts of Cyperus rotundus and Struthanthus flexicaulis were more effective in inhibiting the efflux systems. Great variety of complex phenolic compounds, including flavonoids, was observed in these plant extracts, which may be involved in the biological activities. Despite low number of chicken carcasses positive for Campylobacter spp. observed in this study, identification of high levels of antimicrobial resistance is an important problem and emphasizes the need for continuous monitoring and new ways to control this pathogen in the poultry production chain. The results reported here suggest that plant extracts may be used to control Campylobacter spp. in chicken and by- products by reducing of the poultry colonization. Nevertheless, future studies are required for more detailed characterization of plant extracts, isolation and identification of active principles, as well as to evaluate activity of these plant extracts in vivo. |
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Hungaro, Humberto Moreirahttp://lattes.cnpq.br/8309143011476182Moreira, Maria Aparecida Scatamburlohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797678J6Leite, João Paulo Vianahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763897U8Mendonça, Regina Célia Santoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790986E3Cunha, Luciana Rodrigues dahttp://lattes.cnpq.br/3546793877060403Pinto, Cláudia Lúcia de Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783521J62015-03-26T12:25:14Z2015-01-202015-03-26T12:25:14Z2014-02-20HUNGARO, Humberto Moreira. Activity of plant extracts on multidrug efflux systems from antimicrobial multi-resistant Campylobacter spp.. 2014. 102 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2014.http://locus.ufv.br/handle/123456789/490The high occurrence of Campylobacter spp. in chicken meat and by- products and their increasing antimicrobial resistance have worried public health authorities worldwide. The objectives of this study were to evaluate the Campylobacter contamination in chicken carcasses and antimicrobial resistance of the isolates, as well as to investigate the resistance-modifying and efflux inhibitory activities of plant extract against this microorganism. A total of 95 chicken carcasses from slaughterhouses from Minas Gerais Brazil were analyzed by most probable number method (MPN) associated with subsequent confirmation by polymerase chain reaction (PCR). The isolates were screened for antimicrobial resistance by agar diffusion disk method, and the main genes and mechanisms of resistance were investigated. Twenty ethanolic extracts of plants were tested for antimicrobial, resistance-modifying and accumulation of ethidium bromide activities. The major classes of secondary metabolites in the plant extracts showing biological activities were analyzed by high performance liquid chromatography (HPLC). Campylobacter spp. was found in 16.8% of samples with microbial load ranging from 60.0 to 184 MPN/carcass, and C. jejuni was the species most prevalent (93.8 %). The isolates were resistant to at least 5 (31.2 %) of the antimicrobial agents tested, and high resistance rates were observed for cephalosporins (75 - 100 %), quinolones (95 %), tetracycline (50 %) and ampicillin (45 %). Ciprofloxacin resistance proved to be caused by mutation in the gyrA gene, whereas 40 % of the tetracycline resistance was explained by presence of the tet(O) gene. The blaOXA-61 gene, which encodes for a β-lactamase, was found in both resistant and susceptible isolates for ampicillin. Almost all isolates showed the three genes required to synthesize the CmeABC efflux system. The use of phenylalanine-arginine β-naphthylamide (PAβN), a well-known efflux pump inhibitor, decreased the MIC of ciprofloxacin, tetracycline, ampicillin and bile salts by 2 to 8 fold confirming the activity of this resistance mechanism. These extracts showed weak antimicrobial activity comparing to antibiotics, with MIC values ranging from 0.1 to >16.38 mg/mL. However, most of them showed synergistic effects on combining with bile salts when used at subinhibitory concentration, decreasing 2 to 16 fold the MIC values of these antimicrobials. We observed that ten plant extracts increased the accumulation of ethidium bromide in the Campylobacter cells, suggesting inhibition of efflux systems. The extracts of Alpinia zerumbet and Vernonia polyanthes showed greater synergistic effect on combining with bile salts, whereas extracts of Cyperus rotundus and Struthanthus flexicaulis were more effective in inhibiting the efflux systems. Great variety of complex phenolic compounds, including flavonoids, was observed in these plant extracts, which may be involved in the biological activities. Despite low number of chicken carcasses positive for Campylobacter spp. observed in this study, identification of high levels of antimicrobial resistance is an important problem and emphasizes the need for continuous monitoring and new ways to control this pathogen in the poultry production chain. The results reported here suggest that plant extracts may be used to control Campylobacter spp. in chicken and by- products by reducing of the poultry colonization. Nevertheless, future studies are required for more detailed characterization of plant extracts, isolation and identification of active principles, as well as to evaluate activity of these plant extracts in vivo.A elevada ocorrência de Campylobacter spp. em carne e subprodutos de frango e sua crescente resistência a antimicrobianos têm preocupado as autoridades sanitárias em todo o mundo. Os objetivos deste estudo foram avaliar a contaminação de carcaças de frango por Campylobacter spp. e a resistência a antimicrobianos dos isolados; bem como investigar a capacidade de extratos vegetais em modular a atividade de antimicrobianos e de inibir os sistemas de efluxo expressos por este micro-organismo. Noventa e cinco carcaças de frangos abatidos em Minas Gerais Brasil foram analisadas por meio da técnica de Número Mais Provável (NMP) com confirmação pela reação de polimerase em cadeia (PCR). Os isolados foram submetidos ao teste de resistência a antimicrobianos por disco de difusão em ágar, e os principais genes e mecanismos de resistência foram investigados. Vinte extratos vegetais etanólicos foram avaliados quanto às atividades antimicrobiana, moduladora de resistência e de acumulação de brometo de etídeo. As principais classes de metabólitos secundários de extratos vegetais com maior atividade biológica foram analisadas por cromatografia líquida de alta eficiência (HPLC). Campylobacter spp. foi identificado em 16,8 % das amostras, as contagens variaram entre 60,0 - 184 NMP/carcaça, e a espécie C. jejuni foi prevalente (93.8 %). Os isolados foram resistentes a no mínimo cinco (31,2 %) dos antimicrobianos testados e maior resistência foi observada para cefalosporinas (75 - 100 %), quinolonas (95 %), tetraciclina (50 %) e ampicilina (45 %). A resistência a ciprofloxacina foi associada a mutações no gene gyrA, enquanto que 40 % da resistência à tetraciclina foi relacionada à presença do gene tet(O). O gene blaOXA-61, que codifica uma β-lactamase, foi encontrado em isolados resistentes e suscetíveis a ampicilina. A maioria dos isolados apresentou os três genes do sistema de efluxo CmeABC. O uso de fenilalanina-arginil- naftilamida (PAβN), um inibidor de sistemas de efluxo, diminuiu a MIC de ciprofloxacina, tetraciclina, ampicilina e sais biliares entre 2 e 128 vezes, o que confirmou a atividade deste mecanismo de resistência. Os extratos vegetais apresentaram fraca atividade antimicrobiana em comparação com os antibióticos, com a MIC variando entre 0,1 e >16,38 mg/mL. Entretanto, a maioria deles apresentou sinergismo em combinação com sais biliares quando utilizados em concentrações subinibitórias, com diminuição da MIC destes antimicrobianos entre 2 e 16 vezes. Observou-se que dez extratos vegetais aumentaram a acumulação de brometo de etídeo por Campylobacter spp., o que indica a inibição de sistemas de efluxo. Os extratos de Alpinia zerumbet e Vernonia polyanthes apresentaram maior efeito sinérgico com sais biliares, enquanto que os extratos de Cyperus rotundus e Struthanthus flexicaulis foram mais eficazes na inibição dos sistemas de efluxo. Grande variedade de compostos fenólicos, principalmente flavonóides, foi observada nestes extratos vegetais, os quais podem estar envolvidos nas atividades biológicas. Apesar do baixo número de carcaças de frango positivas para Campylobacter spp. observado neste estudo, a identificação de altos níveis de resistência a antimicrobianos é um importante problema e enfatiza a necessidade de monitoramento contínuo e de novas formas de controle deste patógeno na cadeia produtiva de aves. Os resultados aqui reportados sugerem que extratos vegetais podem ser utilizados para controlar Campylobacter spp. em carne de frango e subprodutos por meio da redução da colonização das aves. Contudo, existe a necessidade de trabalhos futuros para caracterização mais detalhada dos extratos vegetais, isolamento e identificação dos princípios ativos, bem como estudos para avaliar sua atividade in vivo.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaDoutorado em Ciência e Tecnologia de AlimentosUFVBRCiência de Alimentos; Tecnologia de Alimentos; Engenharia de AlimentosBomba de efluxoExtratos vegetaisCampylobacter spp.Resistência antimicrobianaEfflux pumpPlant extractsCampylobacter spp .Antimicrobial resistanceCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOSAtividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianosActivity of plant extracts on multidrug efflux systems from antimicrobial multi-resistant Campylobacter spp.info: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/pdf1352868https://locus.ufv.br//bitstream/123456789/490/1/texto%20completo.pdf30b34da64bf56493e7800cca82638153MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain219006https://locus.ufv.br//bitstream/123456789/490/2/texto%20completo.pdf.txt7e298effdbebcef254ff33769652702cMD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3705https://locus.ufv.br//bitstream/123456789/490/3/texto%20completo.pdf.jpgcb943b5ffc30758ad92b878aaf5e83d2MD53123456789/4902016-04-06 23:07:43.262oai:locus.ufv.br:123456789/490Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-07T02:07:43LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
dc.title.alternative.eng.fl_str_mv |
Activity of plant extracts on multidrug efflux systems from antimicrobial multi-resistant Campylobacter spp. |
title |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
spellingShingle |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos Hungaro, Humberto Moreira Bomba de efluxo Extratos vegetais Campylobacter spp. Resistência antimicrobiana Efflux pump Plant extracts Campylobacter spp . Antimicrobial resistance CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS |
title_short |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
title_full |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
title_fullStr |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
title_full_unstemmed |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
title_sort |
Atividade de extratos vegetais sobre sistema de efluxo multidrogas em Campylobacter spp. multirresistentes a antimicrobianos |
author |
Hungaro, Humberto Moreira |
author_facet |
Hungaro, Humberto Moreira |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/8309143011476182 |
dc.contributor.author.fl_str_mv |
Hungaro, Humberto Moreira |
dc.contributor.advisor-co1.fl_str_mv |
Moreira, Maria Aparecida Scatamburlo |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797678J6 |
dc.contributor.advisor-co2.fl_str_mv |
Leite, João Paulo Viana |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763897U8 |
dc.contributor.advisor1.fl_str_mv |
Mendonça, Regina Célia Santos |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4790986E3 |
dc.contributor.referee1.fl_str_mv |
Cunha, Luciana Rodrigues da |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/3546793877060403 |
dc.contributor.referee2.fl_str_mv |
Pinto, Cláudia Lúcia de Oliveira |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783521J6 |
contributor_str_mv |
Moreira, Maria Aparecida Scatamburlo Leite, João Paulo Viana Mendonça, Regina Célia Santos Cunha, Luciana Rodrigues da Pinto, Cláudia Lúcia de Oliveira |
dc.subject.por.fl_str_mv |
Bomba de efluxo Extratos vegetais Campylobacter spp. Resistência antimicrobiana |
topic |
Bomba de efluxo Extratos vegetais Campylobacter spp. Resistência antimicrobiana Efflux pump Plant extracts Campylobacter spp . Antimicrobial resistance CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS |
dc.subject.eng.fl_str_mv |
Efflux pump Plant extracts Campylobacter spp . Antimicrobial resistance |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS::CIENCIA DE ALIMENTOS |
description |
The high occurrence of Campylobacter spp. in chicken meat and by- products and their increasing antimicrobial resistance have worried public health authorities worldwide. The objectives of this study were to evaluate the Campylobacter contamination in chicken carcasses and antimicrobial resistance of the isolates, as well as to investigate the resistance-modifying and efflux inhibitory activities of plant extract against this microorganism. A total of 95 chicken carcasses from slaughterhouses from Minas Gerais Brazil were analyzed by most probable number method (MPN) associated with subsequent confirmation by polymerase chain reaction (PCR). The isolates were screened for antimicrobial resistance by agar diffusion disk method, and the main genes and mechanisms of resistance were investigated. Twenty ethanolic extracts of plants were tested for antimicrobial, resistance-modifying and accumulation of ethidium bromide activities. The major classes of secondary metabolites in the plant extracts showing biological activities were analyzed by high performance liquid chromatography (HPLC). Campylobacter spp. was found in 16.8% of samples with microbial load ranging from 60.0 to 184 MPN/carcass, and C. jejuni was the species most prevalent (93.8 %). The isolates were resistant to at least 5 (31.2 %) of the antimicrobial agents tested, and high resistance rates were observed for cephalosporins (75 - 100 %), quinolones (95 %), tetracycline (50 %) and ampicillin (45 %). Ciprofloxacin resistance proved to be caused by mutation in the gyrA gene, whereas 40 % of the tetracycline resistance was explained by presence of the tet(O) gene. The blaOXA-61 gene, which encodes for a β-lactamase, was found in both resistant and susceptible isolates for ampicillin. Almost all isolates showed the three genes required to synthesize the CmeABC efflux system. The use of phenylalanine-arginine β-naphthylamide (PAβN), a well-known efflux pump inhibitor, decreased the MIC of ciprofloxacin, tetracycline, ampicillin and bile salts by 2 to 8 fold confirming the activity of this resistance mechanism. These extracts showed weak antimicrobial activity comparing to antibiotics, with MIC values ranging from 0.1 to >16.38 mg/mL. However, most of them showed synergistic effects on combining with bile salts when used at subinhibitory concentration, decreasing 2 to 16 fold the MIC values of these antimicrobials. We observed that ten plant extracts increased the accumulation of ethidium bromide in the Campylobacter cells, suggesting inhibition of efflux systems. The extracts of Alpinia zerumbet and Vernonia polyanthes showed greater synergistic effect on combining with bile salts, whereas extracts of Cyperus rotundus and Struthanthus flexicaulis were more effective in inhibiting the efflux systems. Great variety of complex phenolic compounds, including flavonoids, was observed in these plant extracts, which may be involved in the biological activities. Despite low number of chicken carcasses positive for Campylobacter spp. observed in this study, identification of high levels of antimicrobial resistance is an important problem and emphasizes the need for continuous monitoring and new ways to control this pathogen in the poultry production chain. The results reported here suggest that plant extracts may be used to control Campylobacter spp. in chicken and by- products by reducing of the poultry colonization. Nevertheless, future studies are required for more detailed characterization of plant extracts, isolation and identification of active principles, as well as to evaluate activity of these plant extracts in vivo. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-02-20 |
dc.date.accessioned.fl_str_mv |
2015-03-26T12:25:14Z |
dc.date.available.fl_str_mv |
2015-01-20 2015-03-26T12:25:14Z |
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 |
HUNGARO, Humberto Moreira. Activity of plant extracts on multidrug efflux systems from antimicrobial multi-resistant Campylobacter spp.. 2014. 102 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2014. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/490 |
identifier_str_mv |
HUNGARO, Humberto Moreira. Activity of plant extracts on multidrug efflux systems from antimicrobial multi-resistant Campylobacter spp.. 2014. 102 f. Tese (Doutorado em Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos) - Universidade Federal de Viçosa, Viçosa, 2014. |
url |
http://locus.ufv.br/handle/123456789/490 |
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por |
language |
por |
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Universidade Federal de Viçosa |
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Doutorado em Ciência e Tecnologia de Alimentos |
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UFV |
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BR |
dc.publisher.department.fl_str_mv |
Ciência de Alimentos; Tecnologia de Alimentos; Engenharia de Alimentos |
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Universidade Federal de Viçosa |
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