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Previous issue date: 2014-05-19 / Multi-drug resistant Klebsiella pneumoniae that produce the enzyme K. pneumoniae carbapenemase (KPC) are becoming a common cause of infections in health care centers. Furthermore, Klebsiella can develop multicellular biofilms, which lead to elevated adaptive antibiotic resistance. Here, it was described the antimicrobial and anti-biofilm activities of synthetic cationic peptides against K. pneumoniae strains susceptible and carbapenens resistant through in vitro and in vivo assays. By using static microplate assays, it was observed that the concentration of the peptides IDR-1018, DJK-5 and DJK-6 required to prevent biofilm formation by these clinical isolates was below the minimum inhibitory concentration (MIC) of these peptides. Flow cell experiments confirmed the anti-biofilm activity of the peptides against 2 day-old biofilms of different KPC producing isolates and, in some cases, the peptides induced biofilm cell death. Combinations of DJK-6 together with ??-lactam antibiotics, including the carbapenem meropenem, also prevented planktonic growth and biofilm formation of KPC producing strain 1825971. Interestingly, the peptide DJK-6 was able to enhance, by at least 16-fold, the ability of meropenem to eradicate pre-formed biofilms formed by this strain. Although, this combination between meropenem, DJK-6 was effective in vitro, any reduction of bacterial load was observed in murine lung model of infection. Similarly, the peptides HHC-10, IDR-1018 e IDR-1002 was effective in vitro, but ineffective in vivo. The results in vivo, using the peptides HHC-10 and IDR-1018 after infection were contrasting (significant reduction or non-reduction of bacterial load) between two models of lung infection. Others approaches using the peptides IDR-1018 prophylactically or the peptide IDR-1002 (effective in inhibit bacterial growth) before the induction of the infection, as a preventive approach or IDR-1002 (peptide that reduced the bacterial counts in vitro) after infection, also was unable to reduce the bacterial load in lungs of mice. Despite ineffective in acute model of lung infection, the use of cationic peptides, such DJK-6, to potentiate the activity of ??-lactams including meropenem, represents a promising strategy to prevent biofilm formation or eliminate existent biofilms both in medical devices and in body surfaces. / Klebsiella pneumoniae resistente a m??ltiplos antibi??ticos, que produzem a enzima K. pneumoniae carbapenemase (KPC), est??o se tornando uma causa comum de infec????es em centros de cuidado a sa??de. Al??m disso, K. pneumoniae pode desenvolver biofilmes multicelulares, que levam o aumento da resist??ncia adaptativa a antibi??ticos. Aqui foi descrito a atividade antimicrobiana e antibiofilme de pept??deos cati??nicos sint??ticos contra isolados de K. pneumoniae suscept??vel e resistente a carbapenens em ensaios in vitro e in vivo. Usando ensaios de microplaca est??tico, foi observado que a concentra????o dos pept??deos IDR-1018, DJK-5 e DJK-6, requerida para prevenir a forma????o de biofilmes de isolados resistentes foi abaixo da concentra????o inibit??ria m??nima (CIM) desses pept??deos. Experimentos de fluxo de c??lula confirmaram a atividade antibiofilme dos pept??deos contra biofilmes pr??-formados de diferentes isolados de K. pneumoniae produtores de KPC e, em alguns casos, os pept??deos induziram morte celular. Combina????es de DJK-6 e antibi??ticos ??-lact??micos, incluindo o carbapenem meropenem, tamb??m preveniram o crescimento planct??nico e a forma????o de biofilmes do isolado produtor de KPC 1825971. Interessantemente, o pept??deo DJK-6 foi capaz de aumentar, em pelo menos 16 vezes, a habilidade de meropenem erradicar biofilmes pr??-formados desse isolado. Embora, essa combina????o tenha sido efetiva in vitro, nenhuma redu????o da carga bacteriana foi observada em modelo murino de infec????o pulmonar por K. pneumoniae. Os resultados dos pept??deos HHC-10 e IDR-1018 (os quais inibiram o crescimento bacteriano in vitro a concentra????o que apresentou baixa citotoxicidade) foram contrastantes entre os dois modelos de infec????o pulmonar (significando redu????o ou n??o redu????o da carga bacteriana). Outras abordagens utilizando os pept??deos IDR-1018 profilaticamente ou o pept??deo IDR-1002 (efetivo em inibir o crescimento bacteriano in vitro) ap??s a infec????o, tamb??m foram incapazes de reduzir a carga bacteriana em pulm??es de camundongos. Apesar da inefetividade em modelo agudo de infec????o pulmonar, o uso de pept??deos cati??nicos, tal como DJK-6, para potencializar a atividade de ??-lact??micos, incluindo meropenem, representa uma estrat??gia promissora para prevenir a forma????o de biofilmes ou eliminar biofilmes existentes tanto em equipamentos m??dicos quanto em superf??cies do corpo.
| Identifer | oai:union.ndltd.org:IBICT/oai:bdtd.ucb.br:tede/1996 |
| Date | 19 May 2014 |
| Creators | Ribeiro, Suzana Meira |
| Contributors | Franco, Oct??vio Luiz, Moreno, Susana Elisa |
| Publisher | Universidade Cat??lica de Bras??lia, Programa Strictu Sensu em Ci??ncias Gen??micas e Biotecnologia, UCB, Brasil, Escola de Sa??de e Medicina |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UCB, instname:Universidade Católica de Brasília, instacron:UCB |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | -2827197273900952156, 500, 500, 600, -6392058866414562720, -5518144268585252051 |
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