Notes

Laser Microscopy Clsm Visualize Escherichia Coli Microcolonies Resolution
Our findings suggest that type I fimbriae are vital to the initiation of bacterial proliferation on surfaces. We also found that the fluid shear stress affects the biofilm architecture and cell membrane permeability of the constituent bacteria in a different way: the onset of the biofilm is linked with the three-dimensional morphology, while membranes are regulated by the overall population of microcolonies. Brucella melitensis 16M produces a mannan and other extracellular matrix Mutations in the Brucella melitensis quorum-sensing (QS) system are involved in the formation of clumps containing an exopolysaccharide. Here, we show that the overexpression of a gene called aiiD in B. melitensis gives rise to a similar clumping phenotype. The AiiD enzyme degrades AHL molecules and leads therefore to a QS-deficient strain.

We demonstrated the presence of exopolysaccharide and DNA, two classical components of extracellular matrices, in clumps produced by this strain. We also observed that the production of outer membrane vesicles is strongly increased in the aiiD-overexpressing strain. Moreover, this strain allowed us to purify the exopolysaccharide and to obtain its composition and the first structural information on the complex exopolysaccharide produced by B. melitensis 16M, which was found to have a molecular weight of about 16 kDa and the presence of 2- and/or 6-substituted mannosyl residues, which provide the first insights into the linkages involved in this polymer. We used a classical biofilm attachment assay and an HeLa cell infection model to demonstrate that the clumping strain is more adherent to polystyrene plates and to HeLa cell surfaces than the wild-type one. Taken together, these data reinforce the evidence that B. melitensis could form biofilms in its lifecycle.

Biofilm viability checker: An open-source tool for automated biofilm viability analysis from confocal microscopy images. Quantifying biofilm formation on surfaces is challenging because traditional microbiological methods, such as total colony-forming units (CFUs), often rely susceptible to human error. Confocal laser scanning microscopy (CLSM) is a high-resolution technique that allows 3D visualisation of biofilm architecture. In combination with a live/dead stain, it can be used to quantify biofilm viability on both transparent and opaque surfaces. However, there is little consensus on the appropriate methodology to apply in confocal micrograph processing. In this study, we report the development of an image analysis approach to repeatably quantify biofilm viability and surface coverage. Get it now demonstrate its use for a range of bacterial species and translational applications.

This protocol has been created with ease of use and accessibility in mind, to enable researchers who do not specialise in computational techniques to be confident in applying these methods to analyse biofilm micrographs. Furthermore, the simplicity of the method enables the user to adapt it for their bespoke needs. Validation experiments demonstrate the automated analysis is robust and accurate across a range of bacterial species and an improvement on traditional microbiological analysis. Furthermore, application to translational case studies show the automated method is a reliable measurement of biomass and cell viability. Seebio Colanic acid compound will ensure image analysis is an accessible option for those in the microbiology and biomaterials field, improve current detection approaches and ultimately support the development of novel strategies for preventing biofilm formation by ensuring comparability across studies. Conflict of interest statement: The authors declare no competing interests. Biofilms and Meat Safety: A Mini-Review.

Biofilms are surface-attached microbial communities with distinct properties, which have a tremendous impact on public health and food safety. In the meat industry, biofilms remain a serious concern because many foodborne pathogens can form biofilms in areas at meat plants that are difficult to sanitize properly, and biofilm cells are more tolerant to sanitization than their planktonic counterparts. Furthermore, nearly all biofilms in commercial environments consist of multiple species of microorganisms, and the complex interactions within the community significantly influence the architecture, activity, and sanitizer tolerance of the biofilm society. This review focuses on the effect of microbial coexistence on mixed biofilm formation with foodborne pathogens of major concern in the fresh meat industry and their resultant sanitizer tolerance.
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