Mucus organisation is shaped by colonic content; a new view.

The colonic mucus barrier is commonly described as a continuous double layer covering the epithelium, separating the microbiota from the intestinal tissue. This model is currently considered valid throughout the colon. The colon is characterised by regional anatomo-functional specificities such as presence and consistency of contents and location. In this study, we characterised the organisation of the colonic mucus barrier in proximal and distal colon of rodents by histological and FISH staining, taking into account aforementioned specificities. By using longitudinal sections and imaging extensive areas of tissue with and without colonic contents, we have obtained a spatiotemporal overview of mucus organisation in the colon. We describe for the first time that the colonic mucus layer covers the faeces instead of the epithelium in the distal colon. This faecal mucus layer confines the microbiota to the faeces and prevents it from remaining in empty distal colon. In the proximal colon, the mucus did not form a separating layer between bacteria and epithelium. We conclude that the organisation of colonic mucus is reliant on the presence of the colonic content, and the location within the colon. Our findings reopen the discussion on the nature of the colonic mucus barrier.

Transmission electron microscopy for elucidating the impact of silver-based treatments (ionic silver versus nanosilver-containing coating) on the model yeast Saccharomyces cerevisiae.

After exposure to ionic silver or nanosilver-containing plasma coating, the same visual aspect of scanning transmission electron microscopy (STEM) images was observed for the model yeast Saccharomyces cerevisiae. The main common feature was the presence of electron-dense nodules all over the cell. However, high resolution TEM (HRTEM), STEM, energy dispersive x-ray microanalysis spectroscopy (EDS) and electron microdiffraction revealed some striking differences. Regarding ionic silver exposure, the formation of electron-dense nodules was related to the Ag(+) reactivity towards sulfur-containing compounds to form clusters with Ag(2)S-like structures, together with the production of a few silver nanocrystals, mainly at the cell wall periphery. For nanosilver-based treatment, some sulfur-containing silver clusters preferentially located at the cell wall periphery were detected, together with nodules composed of silver, sulfur and phosphorus all over the cell. In both silver-based treatments, nitrogen and silver signals overlapped, confirming the affinity of silver entities for proteinaceous compounds. Moreover, in the case of nanosilver, interactions of silver with phosphorus-containing subcellular structures were indicated.

Orientation and detachment dynamics of Bacillus spores from stainless steel under controlled shear flow: modelling of the adhesion force.

Shear-flow induced spore detachment was performed under well-controlled laminar flow conditions, in a specially-designed shear stress flow chamber. By comparing detachment profiles of a panel of four strains, belonging to the B. cereus group (B. cereus and B. thuringiensis) and to less related Bacillus species (B. pumilus), it was shown that the spore ability of attaching to stainless steel, probed under dynamic conditions, was mainly affected by the presence (and number) of appendages. Adhesion force between the B. cereus 98/4 strain and stainless steel was quantified at nanoscale. To this aim, detachment results were combined with a theoretical modelling, based on the balance of hydrodynamic forces and torque exerted over a simplified spore model with a spherical form. The wall shear stress, required to remove 50% of the spores initially attached to stainless steel, was determined. On this basis, an adhesion force of 930 ± 390 pN was obtained. Real-time re-orientation of B. cereus 98/4 spores was experimentally established, by using a high-speed camera for tracking the motions of individual spores with high temporal and spatial resolution. Even though tethered to stainless steel without any detachment occurring, spores kept mobile on the substratum, probably due to the existence of discrete bonds or local clusters of anchoring sites, and tended to re-orientate in the flow direction, for minimizing hydrodynamic forces and torque exerted by fluid flow. A significant heterogeneity within the population was also observed, with the co-existence of both moving and immobile spores.

Synchrotron FTIR microspectroscopy of the yeast Saccharomyces cerevisiae after exposure to plasma-deposited nanosilver-containing coating.

The present work was focused on elucidating changes in the model yeast Saccharomyces cerevisiae (cell composition, ultrastructure) after exposure to antimicrobial plasma-mediated nanocomposite films. In order to achieve this, a nanosilver-containing coating was deposited onto stainless steel using radiofrequency HMDSO plasma deposition, combined with simultaneous silver sputtering. X-ray photoelectron spectroscopy (XPS) confirmed the presence of silver nanoparticles embedded in an organosilicon matrix. In addition, scanning electron microscopy (SEM) demonstrated the nanoparticle-based morphology of the deposited layer. The antifungal properties towards S. cerevisiae were established, since a 1.4 log reduction in viable counts was observed after a 24-h adhesion compared to control conditions with the matrix alone. Differences in cell composition after exposure to the nanosilver was assessed for the protein region using, for the first time, synchrotron Fourier-transform infrared (FTIR) microspectroscopy of single S. cerevisiae cells, through in situ mapping with sub-cellular spatial resolution. IR spectrum of yeast cells recovered after a 24-h adhesion to the nanosilver-containing coating revealed a significant downshift (20 cm(-1)) of the amide I peak at 1655 cm(-1), compared to freshly harvested cells. This lower band position, corresponding to a loss in alpha-helix structures, is indicative of the disordered secondary structures of proteins, due to the transition between active and inactive conformations under nanosilver-induced stress conditions. No significant effect on the nucleic acid region was detected. The inhibitory action of silver was targeted against both cell wall and intracellular proteins such as enzymes. Transmission electron microscopy (TEM) observations of the yeast ultrastructure confirmed serious morphological and structural damages. A homogeneous protein-binding distribution of nanosilver all over the cell was assumed, since the presence of electron-dense silver clusters was detected not only on the cell surface but also within the cell. For control experiments with the organosilicon matrix alone, no antimicrobial effect was observed, which was consistent with synchrotron FTIR results and TEM observations.

Study of bioadhesion on a flat plate with a yeast/glass model system.

The attachment of microorganisms to a surface is a critical first step of biofilm fouling in membrane processes. The shear-induced detachment of baker's yeast in adhesive contact with a plane glass surface was thus experimentally studied, using a specially designed shear stress flow chamber. The yeast was marketed either as rod-shaped pellets (type I yeast) or as spherical pellets (type II yeast). A complete series of experiments for measuring the shear stress necessary to detach a given proportion of individual yeast cells of type I or II was performed under different environmental conditions (ionic strength, contact time). In parallel, the surface physicochemical properties of the cells (surface charge, hydrophobicity, and electron donor and electron acceptor components) were determined. For the first type of yeast cells, which were rather hydrophilic, adhesion to the glass plate was weak. This was due to both electrostatic effects and hydrophilic repulsion. Furthermore, adhesion was not sensitive to any variation of the ionic strength. For yeast of the second type, adhesion was drastically increased. This could be explained by their physicochemical surface properties and especially their hydrophobic and electron acceptor components, which caused strong attractive van der Waals and Lewis acid-base interactions, counterbalancing the electrostatic repulsion. For increasing ionic strengths, adhesion was greater, due to lower electrostatic repulsion. The results were quantified through the definition of a critical wall shear stress ( tau w 50% ) required to detach 50% of the yeast cells initially deposited on the glass surface. The influence of the contact time was also evaluated and it was shown that, whatever the type of yeast, macromolecules such as proteins were released into the extracellular medium due to cell lysis and could contribute to the formation of a conditioning film. As a result, the cells were more strongly stuck to the glass plate.

Novel membrane bioreactor with gas/liquid two-phase flow for high-performance degradation of phenol.

The use of a membrane bioreactor with cell retention to achieve high biomass concentrations has been examined for phenol degradation by the bacteria Alcaligenes eutrophus. This process is particularly interesting for toxic substrates as the hydraulic dilution rate and the growth rate are independently controlled. In the case of a transitory excess of phenol, this potentially toxic situation can be overcome by modifying the substrate concentration or the dilution rate without any loss of cells. The injection of a gas phase at the filter inlet increased both the permeate flow rate (by a factor of 1. 75) and the oxygen transfer capacity (by a factor of 1.5). This has enabled the cell concentration to reach a maximal value of 60 g L-1 with a hydraulic dilution rate of 0.5 h-1 and a phenol feed concentration of 8 g L-1. The volumetric productivity of this process corresponds to a phenol degradation rate approaching 100 kg m-3 day-1. The on-line measurement of the characteristic yellow color of 2-hydroxymuconate semialdehyde, a metabolic intermediate of the phenol degradation pathway, in the permeate provides an interesting basis for process control of phenol supply into the reactor since the color intensity correlates directly to the specific rate of phenol degradation.