Biofilm represents a major public health concern. It is a highly structured and heterogeneous microbial population that is well protected by a hydrated extracellular matrix. In most cases, the difficulties in combating a wide spectrum of biofilm-associated diseases are due to the presence of dormant cells and differential molecular expression. Proteomics is the large-scale and systematic study of cellular proteome expression at any given time by mass spectrometry. It allows high-sensitivity and high-specificity identification of differentially expressed proteins in the biofilms. Over the past few decades, multiple lines of proteomic works have successfully elucidated various aspects of the biofilm including developmental stages, antimicrobial resistance, and survival mechanisms. However, the heterogeneity of biofilms may contribute to inconsistent proteome expression throughout a proteomic experiment. This is due to the fact that the mature biofilm is often associated with the mixture between monolayer and multilayer biofilms, thick microbial population, and chemical gradient of nutrients. This review highlights the biofilm heterogeneities, the principle of mass spectrometry in proteomics, and the possible strategies for quantitative proteomic analysis of heterogeneous biofilms. It is suggested that isolation of monolayer biofilm, laser capture microdissection, flow cytometry, and subtractive proteome profiling may be considered for an accurate and reliable quantitative proteomics experiment.
The present work was performed to determine the impacts of commercial disinfectants against biomass, viability, and biochemical composition of Salmonella enterica serovar Thyphimurium ATCC14028 biofilm. Salmonella Thyphimurium biofilm grown in microplates was exposed to commercial disinfectants namely sodium hypochlorite, benzalkonium chloride, chloroxylenol, and sodium dodecyl-benzene sulfonate-based disinfectants. Biofilm biomass, biofilm viability, and biochemical composition of the biofilm were determined using crystal violet assay, resazurin assay and Fourier transform infrared (FTIR) spectroscopy respectively. Results demonstrated that, among four commercial disinfectants, chloroxylenol-based disinfectant showed the highest inhibition against S. Thyphimurium biofilm. It remarkably hindered biofilm biomass and biofilm viability at all tested concentrations (0.78%-25%). Half-maximal biofilm inhibitory concentration (BIC50) of chloroxylenol-based disinfectant (5.06%) was found to be the lowest among the tested disinfectants. Meanwhile, S. Thyphimurium biofilm treated with chloroxylenol-based disinfectant exhibited changes in FTIR spectral peaks associated with lipid (1460 cm-1), protein (630 cm-1, 702 cm-1, 1550 cm-1 & 1650 cm-1), and nucleic acid (1080 cm-1 & 1229 cm-1). The findings of the present study suggest that the inhibition of chloroxylenol-based disinfectant against S. Thyphimurium biofilm is mediated by structural changes of biofilm.
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