RESUMO
Surface-enhanced Raman scattering (SERS), based on the enhancement of the Raman signal of molecules positioned within a few nanometres from a structured metal surface, is ideally suited to provide bacterial-specific molecular fingerprints which can be used for analytical purposes. However, for some complex structures such as bacteria, the generation of reproducible SERS spectra is still a challenging task. Among the various factors influencing the SERS variability (such as the nature of SERS-active substrate, Raman parameters and bacterial specificity), we demonstrate in this study that the environment of Gram-positive and Gram-negative bacteria deposited on ultra-thin silver films also impacts the origin of the SERS spectra. In the case of densely packed bacteria, the obtained SERS signatures were either characteristic of the secretion of adenosine triphosphate for Staphylococcus aureus (S. aureus) or the cell wall and the pili/flagella for Escherichia coli (E. coli), allowing for an easy discrimination between the various strains. In the case of isolated bacteria, SERS mapping together with principal component analysis revealed some variabilities of the spectra as a function of the bacteria environment and the bactericidal effect of the silver. However, the variability does not preclude the SERS signatures of various E. coli strains to be discriminated.
Assuntos
Escherichia coli/química , Análise Espectral Raman/métodos , Staphylococcus aureus/química , Escherichia coli/citologia , Infecções por Escherichia coli/microbiologia , Humanos , Prata/química , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/citologia , Propriedades de SuperfícieRESUMO
In the last few decades, methicillin-resistant Staphylococcus aureus (MRSA) strains have become a serious health care problem. However, in the European Union/European Economic Area countries the prevalence of the invasive MRSA isolates has decreased in recent years; in Romania, the considerably high prevalence of these strains is still unchanged. In this study, 396 staphylococcal strains were screened using molecular biology techniques for the presence of the nucA, mecA, and mecI genes and for the detection of the possible mutations accumulated in the mecI gene. More than half of the collected Staphylococcus strains (59.34%) were determined as S. aureus, and 63 strains were considered as MRSA. Small number of MRSA strains (n = 6; 54.54% of invasive S. aureus) originated from hemoculture. The mecI gene was present in 22 MRSA strains and in 4 methicillin-resistant coagulase-negative staphylococci strains. The majority of the mecI-positive MRSA strains contained the C to T substitution at position 202; furthermore, one previously undescribed mutation (C to G transversion at nucleotide position 285) was detected in one MRSA strain.
Assuntos
Proteínas de Bactérias/genética , Resistência a Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/genética , Mutação , Proteínas Repressoras/genética , Antibacterianos/uso terapêutico , Técnicas de Tipagem Bacteriana , Hemocultura , Proteínas de Ligação a DNA/genética , Desoxirribonucleases/genética , Expressão Gênica , Humanos , Meticilina/uso terapêutico , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Proteínas de Ligação às Penicilinas/genética , Prevalência , Romênia/epidemiologia , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologiaRESUMO
The approach of combining different mechanisms of antibacterial action by designing hybrid nanomaterials provides a new paradigm in the fight against resistant bacteria. Here, we present a new method for the synthesis of silver nanoparticles enveloped in the biopolymer chitosan. The method aims at the production of bionanocomposites with enhanced antibacterial properties. We find that chitosan and silver nanoparticles act synergistically against two strains of Gram-positive Staphylococcus aureus (S. aureus). As a result the bionanocomposites exhibit higher antibacterial activity than any component acting alone. The minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of the chitosan-silver nanoparticles synthesized at 0 °C were found to be lower than those reported for other types of silver nanoparticles. Atomic force microscopy (AFM) revealed dramatic changes in morphology of S. aureus cells due to disruption of bacterial cell wall integrity after incubation with chitosan-silver nanoparticles. Finally, we demonstrate that silver nanoparticles can be used not only as antibacterial agents but also as excellent plasmonic substrates to identify bacteria and monitor the induced biochemical changes in the bacterial cell wall via surface enhanced Raman scattering (SERS) spectroscopy.
