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In the present study, we investigate the possible contribution of Sphingomonas spp. glycosphingolipids (GSL) and its extracellular polymeric substances (EPS) to the initial colonization and development of biofilm bodies on reverse osmosis (RO) membranes. A combination of an RO cross-flow membrane lab unit, a quartz crystal microbalance with dissipation (QCM-D), and a rear stagnation point flow (RSPF) system with either model bacteria (Sphingomonas wittichii, Escherichia coli, and Pseudomonas aeruginosa) or vesicles made of the bacterial GSL or LPS was used. Results showed noticeable differences in the adhesion LPS versus GSL vesicles in the QCM-D, with the latter exhibiting 50% higher adhesion to polyamide coated crystals (mimicking an RO membrane surface). A similar trend was observed for EPS extracted from S. wittichii, when compared to the adhesion tendency of EPS extracted from P. aeruginosa. By applying the whole-cell approach in the RO lab unit, the cumulative impact of S. wittichii cells composing GSL and probably their EPS reduced the permeate flux during bacterial accumulation on the membrane surface. Experiments were conducted with the same amount of Sphingomonas spp. or Escherichia coli cells resulting in a two times greater flux decline in the presence of S. wittichii. The distinct effects of Sphingomonas spp. on RO membrane biofouling are likely a combination of GSL presence (known for enhancing adhesion when compared to non-GSL containing bacteria) and the EPS contributing to the overall strength of the biofilm matrix.

This investigation is focused on the combined effect of bacterial physicochemical characteristics and motility on cell adhesion and deposition using a flow-through quartz crystal microbalance with dissipation (QCM-D). Three model flagellated strains with different degrees of motility were selected, including a highly motile Escherichia coli K12 MG1655, an environmental strain Sphingomonas wittichii RW1, and a nonmotile (with paralyzed flagella) Escherichia coli K12 MG1655 ΔmotA that is incapable of encoding the motor torque generator for flagellar movement. Of the three strains, S. wittichii RW1 is highly hydrophobic, while E. coli strains are equally hydrophilic. Consideration of the hydrophobicity provides an alternative explanation for the bacterial adhesion behavior. QCM-D results show that motility is a critical factor in determining bacterial adhesion, as long as the aquatic chemical conditions are conducive for motility and the substratum and bacterial surface are similarly hydrophobic or hydrophilic. Once their properties are not similar, the contribution of hydrophobic interactions becomes more pronounced. QCM-D results suggest that during adhesion of the hydrophobic bacterium, S. wittichii RW1, the initial step of adhesion and maturation of bacteria-substratum interaction on hydrophilic surface includes a dynamic change of the viscoelastic properties of the bond bacterium-surface becoming more viscously oriented.

Nonionic surfactants of the form CxEy were studied for their ability to alter the cell surface hydrophobicity and apparent Hamaker constants of a Sphingomonas sp. Through contact angle measurements on hydrated and dried bacterial lawns, it was found that the cell surface hydrophobicity changed systematically with both the alkyl (x) and polyoxyethylene (y) chain lengths. While differences in contact angles were observed between hydrated and dried lawns, they could not be attributed to the mere presence or absence of water, suggesting that reorientation of cell surface components may occur during drying. All surfactants examined reduced the hydrophilicity of the bacterial cell surface, with one surfactant (C18E10) making the cells hydrophobic. Effective bacterial Hamaker constants for bacteria interacting across a vacuum (Abb) and water (Abwb) and bacteria interacting in water with quartz sand (Abws) were calculated from the contact angles. It was found that the surfactants have the potential to reduce the Hamaker constants, but that the overall effects differed between dried and hydrated lawns, indicating that lawn preparation method can have a significant impact in interpretation of cell surface properties. The results also indicate that the Abws value of 10-20 J, which is often assumed in bacterial attachment and transport studies, may be an order of magnitude higher than the actual value. Finally, the results suggest that alteration of bacterial adhesion due to the presence of surfactants cannot be attributed to a single cell surface property but is rather due to multiple interactions.

The adsorption of linear polyoxyethylene (POE) alcohol surfactants of the form CxEy onto the surface of a Sphingomonas sp. has been examined. For this study, the alkyl chain length (x) was fixed at 12 and the POE chain length (y) was varied, with y = 4, 7, 9, 10, and 23 ethylene oxide units. Langmuirian isotherms were observed for C12E4 and C12E23, and more complex isotherms were observed for the three intermediate POE chain length surfactants, with C12E7 and C12E9 exhibiting strong S-shaped isotherms. All isotherms showed plateaus near the critical micelle concentration (CMC) with the plateau decreasing with increasing POE chain length. A simple multi-interaction isotherm is proposed that models the sorption isotherm as the sum of two interactions. The first interaction describes monolayer adsorption, whereas the second interaction describes lateral interactions between sorbed surfactant molecules and the formation of surface aggregates. Varying ratios of these two interactions as a function of POE chain length gives rise to the variety of observed isotherm shapes. Results of the isotherm analysis suggest that lateral interactions dominate for surfactants with low POE chain lengths, and the lateral interactions decrease as the POE chain length is increased.

An investigation of the mobility, viability, and activity of polycyclic aromatic hydrocarbon (PAH) degrading bacteria in an electric field is presented. Bench-scale model aquifers were used to test electrophoresis and electroosmosis as potential mechanisms for bacterial dispersion in contaminated sites. Glass beads, alluvial sand from Lake Geneva, and historically polluted clayey soil were used as packing materials. The green-fluorescent protein labeled PAH-degrading bacteria Sphingomonas sp. L138 and Mycobacterium frederiksbergense LB501TG were used as test organisms because of the known differing physicochemical surface and adhesion properties of the corresponding wild-type strains. No adverse effects of the electric current on bacterial viability and PAH-degradation were observed in the system chosen. Up to 90% of the weakly negatively charged and moderately adhesive cells of strain L138 were transported by electroosmosis, whereas 0-20% were transported by electrophoresis. By contrast, poor electrokinetic transport of strongly charged and highly adhesive cells of M. frederiksbergense LB501TG occurred in the different model aquifers. Treatment of bacteria with the nonionic surfactant Brij35 resulted in up to 80% enhanced electrokinetic dispersion of both strains. Our findings demonstrate that electroosmosis may be a valuable mechanism to transport bacteria in the subsurface with transport efficiencies heavily depending on the retention of the bacteria by the solid phase.

Biodegradable nonionic Tween series surfactants were employed to assess the effects of synthetic surfactants on the bioavailability of a target polycyclic aromatic hydrocarbon (PAH), phenanthrene, in soil/sediment-free micellar solutions. Dosages of surfactants in excess of their respective critical micelle concentrations (CMCs) dramatically enhanced solubilization of phenanthrene, but the micellar-solubilized phenanthrene was neither directly nor readily bioavailable to the PAH-degrading strain, Sphingomonas paucimobilis EPA 505, used in these bioavailability experiments. The microorganism preferred instead to utilize hydrophobic fractions of the Tween surfactants as a carbon source, resulting in an imbalance of amphiphilic moieties in surfactant molecules and associated destabilization of micelles. This effect was assessed by measurements of surface tension, CMCs, weight-based PAH solubilization ratios, and by characterizations of the surfactants via HPLC separation and emulsification behavior. The observations and analyses lead to a conclusion that preferential biological destabilization of surfactant micelles effects an associated release of phenanthrene to the aqueous phase. The phenanthrene so released then apparently reverts to a crystallized form that appears to be bioavailable only through normal re-dissolution to the aqueous phase. This is, to our knowledge, the first attempt to characterize and quantify changes in the properties and solubilization behaviors of surfactant micelles resulting from their partial and preferential biodegradation. The associated re-deposition of previously micellar-solubilized PAHs observed and the loss of solubilization capacity of recovered surfactants have significant implications for applications of surfactant-enhanced bioremediation of contaminated soils and sediments.

The effect of the nonionic surfactant Brij 35 on the bioavailability of solid and Teflon-sorbed dibenzofuran for Sphingomonas sp. strain HH19k was studied in simple model systems. Growth with dibenzofuran and dibenzofuran-specific oxygen uptake in surfactant-free media and with Brij 35 above the critical micelle concentration (cmc) were compared with dissolution and desorption in the absence of bacteria. Brij 35 accelerated dissolution and biodegradation of solid dibenzofuran by a factor of 2. It also enhanced the initial desorption rate of dibenzofuran from Teflon by this factor. Continuously decreasing desorption rates were attributed to slow diffusion of dibenzofuran inside Teflon, leading to depletion of dibenzofuran in the exterior of the Teflon particles. Surprisingly, Brij 35 slowed the initial biodegradation of desorbing dibenzofuran. We propose two processes that led to low bioavailability of sorbed dibenzofuran in the presence of surfactant. First, desorbing dibenzofuran rapidly accumulated in surfactant micelles, leading to reduced truly water-dissolved dibenzofuran concentration as the factor controlling the biodegradation rate. Second, Brij 35 suppressed the contact between bacteria and Teflon. This increased the average diffusion distance of dibenzofuran to the bacteria, which in turn flattened the gradient of the dissolved dibenzofuran concentration between the sorbent and the cells as the driving force for desorption.