Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina

Detalhes bibliográficos
Autor(a) principal: Silva, César Matos Ribeiro da
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFS
Texto Completo: http://ri.ufs.br/jspui/handle/riufs/9789
Resumo: Staphylococcus aureus has been considered a major public health problem worldwide due mainly to the ability to develop resistance to antibiotics. The prevalence of methicillin resistant strains of S. aureus (MRSA) in both hospital and community settings has been increasing, with cases of infections and deaths reported in healthy children and adults. Bacteriocins have been identified as promising alternatives for the control of this pathogen. However, besides the therapeutic use of these peptides still not approved, some studies indicate the possibility of selection of resistant strains. Therefore, prior to the therapeutic use of bacteriocins, studies must be carried out to understand the effect that bacteriocin may have on the selection of resistant strains to avoid the current problem with lineages resistant to traditional antibiotics. In view of the above, the objective of this study was to verify the antimicrobial activity of nisin against strains of MRSA and MSSA isolated from the oropharynx of health professionals and verify the selection of strains resistant to bacteriocin. For the genotypic characterization of the MRSA and MSSA strains, the nucA, LuckPV, mecA and mecC genes were tested. The MSSA lines were positive for amplification of the nucA gene only, confirming the identification of genus and species of these strains. For MRSA strains, the only gene that was not detected was mecC, confirming the methicillin resistance phenotype and that these strains are of human origin and of community environments. The results obtained by the agar diffusion test demonstrated that 80% of the MSSA strains (n = 30) were inhibited by nisin and only one MRSA line (n = 6) showed no sensitivity to bacteriocin. The MIC of MSSA strains ranged from 97.7 to 1250 IU / mL and from MRSA strains of 937.50 to 5000 IU / mL. The DBM for MSSA strains varied from 97.7 IU / mL to values greater than 50,000 IU / mL, and for MRSA, this variation was between 5000 IU / mL at values greater than 10,000 IU / mL depending on the lineage. For most lineages DBM was higher than MIC, showing that the effect of bacteriocin depends on bacteriocin concentration: low concentrations exert bacteriostatic effect and high concentrations bactericidal effect. The addition of increasing concentrations of the bacteriocin to the BHI medium generally resulted in the increase in the lag phase and decrease in the specific growth rate and maximal DO reached by the MSSA and MRSA cultures. MSSA and MRSA strains were transferred for approximately 30 generations in the presence of bacteriocin and a decrease in sensitivity was observed with a consequent increase in nisin MIC for all strains tested. The bacteriocin resistance phenotype has been shown to be a stable trait for these strains and may be associated with a genetic factor. It has also been observed that the use of nisin may trigger cross-resistance to some antibiotics. The results obtained demonstrated that nisin is efficient in controlling the growth of MSSA and MRSA. However, the fact that these lines demonstrate resistance to bacteriocin after transfer in the presence of the same indicates the need to develop strategies to avoid in the future the current problem of resistance to antibiotics. The best way to use bacteriocins therapeutically is to suggest that it is in combination with traditional antibiotics.
id UFS-2_aa62bf5fe924c184ceac8d4d90531bd3
oai_identifier_str oai:ufs.br:riufs/9789
network_acronym_str UFS-2
network_name_str Repositório Institucional da UFS
repository_id_str
spelling Silva, César Matos Ribeiro daBarbosa, Ana Andrea TeixeiraCarneiro, Maria Regina Pires2018-11-22T15:15:09Z2018-11-22T15:15:09Z2018-02-07SILVA, César Matos Ribeiro da. Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina. 2018. 62 f. Dissertação (Mestrado em Biologia Parasitária) - Universidade Federal de Sergipe, São Cristóvão, SE, 2018.http://ri.ufs.br/jspui/handle/riufs/9789Staphylococcus aureus has been considered a major public health problem worldwide due mainly to the ability to develop resistance to antibiotics. The prevalence of methicillin resistant strains of S. aureus (MRSA) in both hospital and community settings has been increasing, with cases of infections and deaths reported in healthy children and adults. Bacteriocins have been identified as promising alternatives for the control of this pathogen. However, besides the therapeutic use of these peptides still not approved, some studies indicate the possibility of selection of resistant strains. Therefore, prior to the therapeutic use of bacteriocins, studies must be carried out to understand the effect that bacteriocin may have on the selection of resistant strains to avoid the current problem with lineages resistant to traditional antibiotics. In view of the above, the objective of this study was to verify the antimicrobial activity of nisin against strains of MRSA and MSSA isolated from the oropharynx of health professionals and verify the selection of strains resistant to bacteriocin. For the genotypic characterization of the MRSA and MSSA strains, the nucA, LuckPV, mecA and mecC genes were tested. The MSSA lines were positive for amplification of the nucA gene only, confirming the identification of genus and species of these strains. For MRSA strains, the only gene that was not detected was mecC, confirming the methicillin resistance phenotype and that these strains are of human origin and of community environments. The results obtained by the agar diffusion test demonstrated that 80% of the MSSA strains (n = 30) were inhibited by nisin and only one MRSA line (n = 6) showed no sensitivity to bacteriocin. The MIC of MSSA strains ranged from 97.7 to 1250 IU / mL and from MRSA strains of 937.50 to 5000 IU / mL. The DBM for MSSA strains varied from 97.7 IU / mL to values greater than 50,000 IU / mL, and for MRSA, this variation was between 5000 IU / mL at values greater than 10,000 IU / mL depending on the lineage. For most lineages DBM was higher than MIC, showing that the effect of bacteriocin depends on bacteriocin concentration: low concentrations exert bacteriostatic effect and high concentrations bactericidal effect. The addition of increasing concentrations of the bacteriocin to the BHI medium generally resulted in the increase in the lag phase and decrease in the specific growth rate and maximal DO reached by the MSSA and MRSA cultures. MSSA and MRSA strains were transferred for approximately 30 generations in the presence of bacteriocin and a decrease in sensitivity was observed with a consequent increase in nisin MIC for all strains tested. The bacteriocin resistance phenotype has been shown to be a stable trait for these strains and may be associated with a genetic factor. It has also been observed that the use of nisin may trigger cross-resistance to some antibiotics. The results obtained demonstrated that nisin is efficient in controlling the growth of MSSA and MRSA. However, the fact that these lines demonstrate resistance to bacteriocin after transfer in the presence of the same indicates the need to develop strategies to avoid in the future the current problem of resistance to antibiotics. The best way to use bacteriocins therapeutically is to suggest that it is in combination with traditional antibiotics.Staphylococcus aureus têm sido considerado um dos maiores problemas de saúde pública a nível mundial devido, principalmente, a habilidade de desenvolver resistência a antibióticos. A prevalência de linhagens de S. aureus resistentes a meticilina (MRSA) tanto em ambiente hospitalar quanto em ambientes comunitários têm aumentado, sendo descritos casos de infecções e relatos de mortes em crianças e adultos saudáveis. Bacteriocinas tem sido apontadas como alternativas promissoras para o controle deste patógeno. Entretanto, além do uso terapêutico destes peptídeos ainda não ser aprovado, alguns estudos indicam a possibilidade de seleção de linhagens resistentes. Portanto, antes do uso terapêutico de bacteriocinas, estudos precisam ser realizados para o entendimento do efeito que a bacteriocina possa ter na seleção de linhagens resistentes para evitar o problema da atualidade com linhagens resistentes aos antibióticos tradicionais. Diante do exposto, o objetivo deste trabalho foi verificar a atividade antimicrobiana da nisina contra cepas de MRSA e MSSA isoladas da orofaringe de profissionais da área de saúde e verificar a seleção de linhagens resistentes à bacteriocina. Para a caracterização genotípica das linhagens MRSA e MSSA, foram testados os genes nucA, LuckPV, mecA e mecC. As linhagens MSSA foram positivas para amplificação apenas do gene nucA, confirmando a identificação de gênero e espécie destas linhagens. Para as linhagens MRSA, o único gene que não foi detectado foi o meC, confirmando o fenótipo de resistência a meticilina e que estas linhagens são de origem humana e de ambientes comunitários. Os resultados obtidos pelo teste de difusão em ágar demonstraram que 80% das linhagens MSSA (n=30) foram inibidas pela nisina e apenas uma linhagem MRSA (n=6) não apresentou sensibilidade à bacteriocina. A CIM das linhagens de MSSA variou de 97,7 a 1250UI/mL e das linhagens MRSA de 937,50 a 5000 UI/mL. A DBM para as linhagens de MSSA variou de 97,7 UI/mL a valores superiores a 50000 UI/mL, e para MRSA esta variação ficou entre 5000 UI/mL a valores maiores que 10000 UI/mL dependendo da linhagem. Para a maioria das linhagens a DBM foi maior que a CIM, mostrando que o efeito da bacteriocina depende da concentração da mesma: baixas concentrações exercem efeito bacteriostático e altas concentrações efeito bactericida. A adição de concentrações crescentes da bacteriocina ao meio BHI resultaram, de maneira geral, no aumento da fase lag e diminuição da velocidade específica de crescimento e DO máxima atingida pelas culturas MSSA e MRSA. Linhagens de MSSA e MRSA foram transferidas por aproximadamente 30 gerações na presença da bacteriocina e foi observada diminuição na sensibilidade com consequente aumento da CIM da nisina para todas as linhagens testadas. O fenótipo de resistência à bacteriocina demonstrou ser uma característica estável para estas linhagens, podendo estar associado a um fator genético. Foi observado também que a utilização da nisina pode desencadear resistência cruzada a alguns antibióticos. Os resultados obtidos demonstraram que a nisina é eficiente em controlar o crescimento de MSSA e MRSA. Entretanto, o fato destas linhagens demonstrarem resistência a bacteriocina após transferência na presença da mesma indica a necessidade de desenvolver estratégias para evitar no futuro o problema atual de resistência a antibióticos. A melhor maneira de usar bacteriocinas terapeuticamente sugere-se que seja em combinação com antibióticos tradicionais.Fundação de Apoio a Pesquisa e à Inovação Tecnológica do Estado de Sergipe - FAPITEC/SESão Cristóvão, SEporBacteriocinasAlternativa terapêuticaResistênciaOrofaringeOropharynxBacteriocinsTherapeutic alternativeResistanceCIENCIAS BIOLOGICAS::PARASITOLOGIAPotencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilinaPotencial of nisin in the control of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive (MSSA)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisPós-Graduação em Biologia ParasitáriaUniversidade Federal de Sergipereponame:Repositório Institucional da UFSinstname:Universidade Federal de Sergipe (UFS)instacron:UFSinfo:eu-repo/semantics/openAccessTEXTCESAR_MATOS_RIBEIRO_SILVA.pdf.txtCESAR_MATOS_RIBEIRO_SILVA.pdf.txtExtracted texttext/plain125510https://ri.ufs.br/jspui/bitstream/riufs/9789/3/CESAR_MATOS_RIBEIRO_SILVA.pdf.txta4c5a986746f0ad34e7a8ca004c8d409MD53THUMBNAILCESAR_MATOS_RIBEIRO_SILVA.pdf.jpgCESAR_MATOS_RIBEIRO_SILVA.pdf.jpgGenerated Thumbnailimage/jpeg1276https://ri.ufs.br/jspui/bitstream/riufs/9789/4/CESAR_MATOS_RIBEIRO_SILVA.pdf.jpg37c6dbef49042358c5b96e6d76321806MD54ORIGINALCESAR_MATOS_RIBEIRO_SILVA.pdfCESAR_MATOS_RIBEIRO_SILVA.pdfapplication/pdf1501451https://ri.ufs.br/jspui/bitstream/riufs/9789/2/CESAR_MATOS_RIBEIRO_SILVA.pdf55b1e466dde0441575d6af04b5dbee0cMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-81475https://ri.ufs.br/jspui/bitstream/riufs/9789/1/license.txt098cbbf65c2c15e1fb2e49c5d306a44cMD51riufs/97892018-11-22 12:15:09.56oai:ufs.br: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Repositório InstitucionalPUBhttps://ri.ufs.br/oai/requestrepositorio@academico.ufs.bropendoar:2018-11-22T15:15:09Repositório Institucional da UFS - Universidade Federal de Sergipe (UFS)false
dc.title.pt_BR.fl_str_mv Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
dc.title.alternative.eng.fl_str_mv Potencial of nisin in the control of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive (MSSA)
title Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
spellingShingle Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
Silva, César Matos Ribeiro da
Bacteriocinas
Alternativa terapêutica
Resistência
Orofaringe
Oropharynx
Bacteriocins
Therapeutic alternative
Resistance
CIENCIAS BIOLOGICAS::PARASITOLOGIA
title_short Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
title_full Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
title_fullStr Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
title_full_unstemmed Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
title_sort Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina
author Silva, César Matos Ribeiro da
author_facet Silva, César Matos Ribeiro da
author_role author
dc.contributor.author.fl_str_mv Silva, César Matos Ribeiro da
dc.contributor.advisor1.fl_str_mv Barbosa, Ana Andrea Teixeira
dc.contributor.advisor-co1.fl_str_mv Carneiro, Maria Regina Pires
contributor_str_mv Barbosa, Ana Andrea Teixeira
Carneiro, Maria Regina Pires
dc.subject.por.fl_str_mv Bacteriocinas
Alternativa terapêutica
Resistência
Orofaringe
Oropharynx
topic Bacteriocinas
Alternativa terapêutica
Resistência
Orofaringe
Oropharynx
Bacteriocins
Therapeutic alternative
Resistance
CIENCIAS BIOLOGICAS::PARASITOLOGIA
dc.subject.eng.fl_str_mv Bacteriocins
Therapeutic alternative
Resistance
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::PARASITOLOGIA
description Staphylococcus aureus has been considered a major public health problem worldwide due mainly to the ability to develop resistance to antibiotics. The prevalence of methicillin resistant strains of S. aureus (MRSA) in both hospital and community settings has been increasing, with cases of infections and deaths reported in healthy children and adults. Bacteriocins have been identified as promising alternatives for the control of this pathogen. However, besides the therapeutic use of these peptides still not approved, some studies indicate the possibility of selection of resistant strains. Therefore, prior to the therapeutic use of bacteriocins, studies must be carried out to understand the effect that bacteriocin may have on the selection of resistant strains to avoid the current problem with lineages resistant to traditional antibiotics. In view of the above, the objective of this study was to verify the antimicrobial activity of nisin against strains of MRSA and MSSA isolated from the oropharynx of health professionals and verify the selection of strains resistant to bacteriocin. For the genotypic characterization of the MRSA and MSSA strains, the nucA, LuckPV, mecA and mecC genes were tested. The MSSA lines were positive for amplification of the nucA gene only, confirming the identification of genus and species of these strains. For MRSA strains, the only gene that was not detected was mecC, confirming the methicillin resistance phenotype and that these strains are of human origin and of community environments. The results obtained by the agar diffusion test demonstrated that 80% of the MSSA strains (n = 30) were inhibited by nisin and only one MRSA line (n = 6) showed no sensitivity to bacteriocin. The MIC of MSSA strains ranged from 97.7 to 1250 IU / mL and from MRSA strains of 937.50 to 5000 IU / mL. The DBM for MSSA strains varied from 97.7 IU / mL to values greater than 50,000 IU / mL, and for MRSA, this variation was between 5000 IU / mL at values greater than 10,000 IU / mL depending on the lineage. For most lineages DBM was higher than MIC, showing that the effect of bacteriocin depends on bacteriocin concentration: low concentrations exert bacteriostatic effect and high concentrations bactericidal effect. The addition of increasing concentrations of the bacteriocin to the BHI medium generally resulted in the increase in the lag phase and decrease in the specific growth rate and maximal DO reached by the MSSA and MRSA cultures. MSSA and MRSA strains were transferred for approximately 30 generations in the presence of bacteriocin and a decrease in sensitivity was observed with a consequent increase in nisin MIC for all strains tested. The bacteriocin resistance phenotype has been shown to be a stable trait for these strains and may be associated with a genetic factor. It has also been observed that the use of nisin may trigger cross-resistance to some antibiotics. The results obtained demonstrated that nisin is efficient in controlling the growth of MSSA and MRSA. However, the fact that these lines demonstrate resistance to bacteriocin after transfer in the presence of the same indicates the need to develop strategies to avoid in the future the current problem of resistance to antibiotics. The best way to use bacteriocins therapeutically is to suggest that it is in combination with traditional antibiotics.
publishDate 2018
dc.date.accessioned.fl_str_mv 2018-11-22T15:15:09Z
dc.date.available.fl_str_mv 2018-11-22T15:15:09Z
dc.date.issued.fl_str_mv 2018-02-07
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv SILVA, César Matos Ribeiro da. Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina. 2018. 62 f. Dissertação (Mestrado em Biologia Parasitária) - Universidade Federal de Sergipe, São Cristóvão, SE, 2018.
dc.identifier.uri.fl_str_mv http://ri.ufs.br/jspui/handle/riufs/9789
identifier_str_mv SILVA, César Matos Ribeiro da. Potencial da nisina no controle de Staphylococcus aureus resistente (MRSA) e sensível (MSSA) à meticilina. 2018. 62 f. Dissertação (Mestrado em Biologia Parasitária) - Universidade Federal de Sergipe, São Cristóvão, SE, 2018.
url http://ri.ufs.br/jspui/handle/riufs/9789
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.program.fl_str_mv Pós-Graduação em Biologia Parasitária
dc.publisher.initials.fl_str_mv Universidade Federal de Sergipe
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFS
instname:Universidade Federal de Sergipe (UFS)
instacron:UFS
instname_str Universidade Federal de Sergipe (UFS)
instacron_str UFS
institution UFS
reponame_str Repositório Institucional da UFS
collection Repositório Institucional da UFS
bitstream.url.fl_str_mv https://ri.ufs.br/jspui/bitstream/riufs/9789/3/CESAR_MATOS_RIBEIRO_SILVA.pdf.txt
https://ri.ufs.br/jspui/bitstream/riufs/9789/4/CESAR_MATOS_RIBEIRO_SILVA.pdf.jpg
https://ri.ufs.br/jspui/bitstream/riufs/9789/2/CESAR_MATOS_RIBEIRO_SILVA.pdf
https://ri.ufs.br/jspui/bitstream/riufs/9789/1/license.txt
bitstream.checksum.fl_str_mv a4c5a986746f0ad34e7a8ca004c8d409
37c6dbef49042358c5b96e6d76321806
55b1e466dde0441575d6af04b5dbee0c
098cbbf65c2c15e1fb2e49c5d306a44c
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
repository.name.fl_str_mv Repositório Institucional da UFS - Universidade Federal de Sergipe (UFS)
repository.mail.fl_str_mv repositorio@academico.ufs.br
_version_ 1802110785367834624