Search

In agroindustry, the hygiene of solid surfaces is of primary importance in order to ensure that products are safe for consumers. To improve safety, one of the major ways consists in identifying and understanding the mechanisms of microbial cell adhesion to nonporous solid surfaces or filtration membranes. In this paper we investigate the adhesion of the yeast cell Saccharomyces cerevisiae (about 5 mum in diameter) to a model solid surface, using well-defined hydrophilic glass substrates. An optical tweezer device developed by Piau [J. Non-Newtonian Fluid Mech. 144, 1 (2007)] was applied to yeast cells in contact with well-characterized glass surfaces. Two planes of observation were used to obtain quantitative measurements of removal forces and to characterize the corresponding mechanisms at a micrometer length scale. The results highlight various adhesion mechanisms, depending on the ionic strength, contact time, and type of yeast. The study has allowed to show a considerable increase of adhering cells with the ionic strength and has provided a quantitative measurement of the detachment forces of cultured yeast cells. Force levels are found to grow with ionic strength and differences in mobility are highlighted. The results clearly underline that a microrheological approach is essential for analyzing the adhesion mechanisms of biological systems at the relevant local scales.

The present study focused on the shear-induced detachment of Saccharomyces cerevisiae in adhesive contact with a 316L stainless steel surface using a shear stress flow chamber, with a view to determining the respective influence of the yeast surface properties and the support characteristics. The effect of cultivation of S. cerevisiae yeast cells on their subsequent detachment from the solid surface was particularly investigated. In order to elucidate the role of stainless steel, non-metallic supports were used as control, covering a broad range of surface properties such as surface free energy and roughness: polypropylene (hydrophobic), polystyrene (mildly hydrophobic, similar to stainless steel) and glass (hydrophilic). All materials were very smooth with respect to the size of yeast. First, experiments were carried out on two types of yeast cells, just rehydrated in saline solution, a biological model widely used in the literature. The influence of the ionic strength (1.5 and 150 mM NaCl) on glass and stainless steel was evaluated. Unlike on glass, no clear evidence was found for electrostatic repulsion with stainless steel since high adhesion was observed whatever the ionic strength. A lack of correlation in adhesion results was also obtained when considering the surface physico-chemical characteristics of type I (hydrophilic) and type II (hydrophobic) rehydrated cells and those of both polymers. It was postulated that unavoidable "sticky" compounds were present on the cell wall, which could not be completely removed during the successive washings of the rehydrated cell suspension before use. This could dramatically alter the yeast surface properties and modify the adhesion strength, thus clearly demonstrating the necessity to work with yeast coming from fresh cultures. Biologically active yeast cells were then used. Once cultured, type I- and type II-yeast cells were shown to exhibit the same hydrophilic properties. Regardless of the material used, for the same ionic strength (150 mM NaCl), yeast adhesion was drastically reduced compared to rehydrated yeast cells. Among all the materials tested, the specificity of 316L stainless steel was clearly established. Indeed, for glass and polymers, cell adhesion was substratum-dependent and driven by the balance between the Lifshitz-van der Waals and Lewis acid/base interactions. Despite nearly identical surface free energies for polystyrene and stainless steel, the metallic surface promoted a totally distinct behaviour which was characterized by a strong - although highly variable - yeast adhesion.

Expanded bed adsorption (EBA) is an interesting primary technology allowing the adsorption of target proteins from unclarified feedstock in order to combine separation, concentration, and purification steps. However, interactions between cells and adsorbent beads during the EBA process can strongly reduce the performance of the separation. So, to minimize these interactions, the mechanisms of cell adsorption on the support were investigated. Adsorption kinetics of the baker's yeast Saccharomyces cerevisiae on the anion exchanger Q Hyper Z were directly performed under real EBA operating conditions, in a lab-scale UpFront 10 column. The yeast was marketed either as rod-shaped pellets (type I yeast) or as spherical pellets (type II yeast). For both types, a complete series of experiments for determining the adsorption profile versus time was performed, varying the superficial velocity or the pH. In parallel, the surface physicochemical properties of the cells (surface charge and electron-donor and electron-acceptor components) and of the support were determined. First of all, whatever the yeast types, the relation between cell adsorption and bed expansion has been highlighted, demonstrating the important role of hydrodynamic. However, for the type II yeast cells, adsorption increased dramatically, compared to the type I, even though it was shown that both types exhibited the same surface charge. In fact, there were strong differences in the Lewis acidic and basic components of the two yeasts. These differences explain the variable affinity toward the support, which was characterized by a strong electron-donor and a weak electron-acceptor component. These observed behaviors agreed with the colloidal theory. This work demonstrates that all kinds of interaction between the cells and the support (electrostatic, Lifshitz-van der Waals, acid/base) have to be taken into account together with hydrodynamic characteristics inside the bed.

The optimal conditions for recovery of an enzyme were determined using gas/liquid two-phase flows. When filtering the enzyme-only solution under single-phase flow conditions, severe fouling occurred. This fouling was manifest as a decline in flux to less than 2% of the initial water flux and a decline in protein concentration in the permeate to 30% of its initial value, during a five-hour filtration period. When yeast cells were added under the same experimental conditions, enzyme transmission was maintained at 100% for the five-hour period and the enzyme mass flux was twofold higher. During gas-sparged microfiltration of the enzyme/yeast mixture in a permeate-recycling mode at the same liquid flow rate, gas/liquid slug flow strongly decreased the transmission of the enzyme (70% decrease), even though the permeate flux was improved (140% improvement). As a result, the mass flux of the enzyme was significantly reduced. However, with a bubble flow pattern, the permeate flux was 1.5 times higher and the transmission was maintained at a high level. The enzyme mass flux was then 25% higher when compared to single-phase flow filtration conditions. During diafiltration experiments with a bubble flow pattern, a 13% higher enzyme recovery was achieved.

The effect of a gas/liquid two-phase flow on the recovery of an enzyme was evaluated and compared with standard crossflow operation when confronted with the microfiltration of a high-fouling yeast suspension. Ceramic tubular and flat sheet membranes were used. At constant feed concentration (permeate recycling) and transmembrane pressure, the results obtained with the tubular membrane were dependent on the two-phase flow pattern. In comparison with single-phase flow performances at the same liquid velocity, the enzyme transmission was maintained at a high level with a bubble flow pattern but it decreased by 70% with a slug flow, whatever the flow rate ratio. Identical results were obtained with flat sheet membranes: for the highest flow rate ratio, the enzyme transmission was reduced by 70% even though the permeate flux was improved by 240%. During diafiltration experiments with the tubular membrane, it was found that a bubble flow pattern led to a 13% higher enzyme recovery compared to single-phase flow conditions, whereas with a slug flow the enzyme recovery was strongly reduced. With bubble flow conditions, energy consumption was minimal, confirming that this flow pattern was the most suitable for enzyme recovery., (Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 610-621, 2002.)

Abstract: The present work focused on the importance of performing EBA experiments under real process conditions. To this end, a high ionic strength medium was used. The influence of cultivating Saccharomyces cerevisiae yeast cells on their subsequent adsorption on a Q HyperZ anion-exchanger was investigated. Two types of industrial yeast cells were used. Once cultured, both types of cells presented similar hydrophilic surface properties and identical adsorption profiles on the anion-exchanger. This was significantly different from the results obtained in previous work on the same yeast cells, just rehydrated in saline buffer, a biological model widely used in the literature. It was postulated that unavoidable “sticky” compounds, initially present in the culture medium or formed during the drying process, were strongly adsorbed on the cell wall and could not be completely removed during the successive washings of the rehydrated cell suspension before use. This could dramatically alter the yeast surface properties and modify the biomass/adsorbent interactions and the bed hydrodynamics, thus demonstrating the necessity to work with yeast coming from fresh cultures. Using biologically active yeast cells allowed to really elucidating the main physico-chemical mechanisms involved in cell adsorption by focusing on the role of the non-electrostatic interactions. Two laboratory mutant yeast strains in which the protein-related cell wall biogenesis was affected, were tested. No significant differences were observed between hydrophilic and hydrophobic yeast cells: the bed remained stable at its initial value and there was a low adsorption ratio in a narrow range between 10% and 17%. This clearly demonstrated that non-electrostatic interactions play a minor role on the affinity of yeast to anion-exchanger adsorbents. [Copyright &y& Elsevier]

Abstract: Composite thin films (∼170nm) containing silver nanoclusters embedded in an organosilicon matrix were deposited by PE-CVD onto stainless steel in order to prevent microbial adhesion. The process originality relies on a dual strategy combining silver sputtering and simultaneous plasma polymerization in argon–hexamethyldisiloxane (HMDSO) plasma, using an asymmetrical RF glow discharge. The metal content in the film was controlled by varying the HMDSO flow rate. Investigation of the physico-chemical properties of the obtained films was conducted by X-ray photoelectron spectroscopy and transmission FTIR spectroscopy. Plasma-mediated coatings were composed of C, O, Si and Ag which was predominantly under metallic form, as indicated by XPS analysis. The presence of Si–H, Si–O–Si, Si–(CH) n –Si and C–H groups was established by FTIR. The yeast Saccharomyces cerevisiae was selected as the model for eukaryotic microorganisms. The maximal anti-adhesive efficiency was achieved for the organosilicon matrix alone. When nanosilver was incorporated into the organic matrix, the efficiency was reduced, especially for high metal contents. Silver antimicrobial property was assumed to be related to Ag+ progressive release from the embedded nanoparticles into the surrounding medium. This release was confirmed by ICP-MS measurements. Moreover, silver-containing film antifungal activity was observed towards sessile cells. [Copyright &y& Elsevier]