47 research outputs found
Protection from Staphylococcus aureus mastitis associated with poly-N-acetyl β-1,6 glucosamine specific antibody production using biofilm-embedded bacteria
Protection from Staphylococcus aureus mastitis associated with poly-N-acetyl beta-1,6 glucosamine specific antibody production using biofilm-embedded bacteria
Staphylococcus aureus vaccines based on bacterins surrounded by slime, surface polysaccharides
coupled to protein carriers and polysaccharides embedded in liposomes administered together with
non-biofilm bacterins confer protection against mastitis. However, it remains unknown whether
protective antibodies are directed to slime-associated known exopolysaccharides and could be
produced in the absence of bacterin immunizations. Here, a sheep mastitis vaccination study was
carried out using bacterins, crude bacterial extracts or a purified exopolysaccharide from biofil
Functional Amyloids Composed of Phenol Soluble Modulins Stabilize Staphylococcus aureus Biofilms
Staphylococcus aureus is an opportunistic pathogen that colonizes the skin and mucosal surfaces of mammals. Persistent staphylococcal infections often involve surface-associated communities called biofilms. Here we report the discovery of a novel extracellular fibril structure that promotes S. aureus biofilm integrity. Biochemical and genetic analysis has revealed that these fibers have amyloid-like properties and consist of small peptides called phenol soluble modulins (PSMs). Mutants unable to produce PSMs were susceptible to biofilm disassembly by matrix degrading enzymes and mechanical stress. Previous work has associated PSMs with biofilm disassembly, and we present data showing that soluble PSM peptides disperse biofilms while polymerized peptides do not. This work suggests the PSMs' aggregation into amyloid fibers modulates their biological activity and role in biofilms
agr-Mediated Dispersal of Staphylococcus aureus Biofilms
The agr quorum-sensing system of Staphylococcus aureus modulates the expression of virulence factors in response to autoinducing peptides (AIPs). Recent studies have suggested a role for the agr system in S. aureus biofilm development, as agr mutants exhibit a high propensity to form biofilms, and cells dispersing from a biofilm have been observed displaying an active agr system. Here, we report that repression of agr is necessary to form a biofilm and that reactivation of agr in established biofilms through AIP addition or glucose depletion triggers detachment. Inhibitory AIP molecules did not induce detachment and an agr mutant was non-responsive, indicating a dependence on a functional, active agr system for dispersal. Biofilm detachment occurred in multiple S. aureus strains possessing divergent agr systems, suggesting it is a general S. aureus phenomenon. Importantly, detachment also restored sensitivity of the dispersed cells to the antibiotic rifampicin. Proteinase K inhibited biofilm formation and dispersed established biofilms, suggesting agr-mediated detachment occurred in an ica-independent manner. Consistent with a protease-mediated mechanism, increased levels of serine proteases were detected in detaching biofilm effluents, and the serine protease inhibitor PMSF reduced the degree of agr-mediated detachment. Through genetic analysis, a double mutant in the agr-regulated Aur metalloprotease and the SplABCDEF serine proteases displayed minimal extracellular protease activity, improved biofilm formation, and a strongly attenuated detachment phenotype. These findings indicate that induction of the agr system in established S. aureus biofilms detaches cells and demonstrate that the dispersal mechanism requires extracellular protease activity
Expression of the multiple resistance operon (mar) during growth of Escherichia coli as a biofilm
An evaluation of the potential role of the multiple antibiotic resistance operon (mar) and the multi-drug efflux pump acrAB in the resistance of E. coli biofilms towards ciprofloxacin
The Biofilm Exopolysaccharide Polysaccharide Intercellular Adhesin—A Molecular and Biochemical Approach
Antibody to a conserved antigenic target is protective against diverse prokaryotic and eukaryotic pathogens
A direct, heavy metal free synthesis of the β-1,6-linked GlcNAc disaccharide
The major component of the matrix of many bacterial biofilms is a linear polymer of β-1,6-linked units of N-acetylglucosamine (polysaccharide intercellular adhesin or poly-N-acetylβ - 1,6-D-glucosamine). In order to facilitate synthetic vaccine construction we have developed a direct, inexpensive, and biologically compatible synthesis of the minimal building block of this polymer, a β -1,6-linked GlcNAc disaccharide, namely 6-O-[2-acetamido-2-deoxy - β-D-glucopyranosyl]-2-acetamido-2-deoxy-D- glucopyranose