Disrupting Irreversible Bacterial Adhesion and Biofilm Formation with an Engineered Enzyme.

Biofilm formation is often attributed to postharvest bacterial persistence on fresh produce and food handling surfaces. In this study, a predicted glycosyl hydrolase enzyme was expressed, purified, and validated for the removal of microbial biofilms from biotic and abiotic surfaces under conditions used for chemical cleaning agents. Crystal violet biofilm staining assays revealed that 0.1 mg/ml of enzyme inhibited up to 41% of biofilm formation by Escherichia coli O157:H7, E. coli 25922, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes. Furthermore, the enzyme was effective at removing mature biofilms, providing a 35% improvement over rinsing with a saline solution alone. Additionally, a parallel-plate flow cell was used to directly observe and quantify the impact of enzyme rinses on E. coli O157:H7 cells adhering to spinach leaf surfaces. The presence of 1 mg/liter enzyme resulted in nearly 6-times-higher detachment rate coefficients than a deionized (DI) water rinse, while the total cells removed from the surface increased from 10% to 25% over the 30-min rinse time, reversing the initial phases of biofilm formation. Enzyme treatment of all 4 cell types resulted in significantly reduced cell surface hydrophobicity and collapse of negatively stained E. coli 25922 cells imaged by electron microscopy, suggesting potential polysaccharide surface modification of enzyme-treated bacteria. Collectively, these results point to the broad substrate specificity and robustness of the enzyme for different types of biofilm stages, solution conditions, and pathogen biofilm types and may be useful as a method for the removal or inhibition of bacterial biofilm formation. IMPORTANCE In this study, the ability of an engineered enzyme to reduce bacterial adhesion and biofilm formation of several foodborne pathogens was demonstrated, representing a promising option for enhancing or replacing chlorine and other chemical sanitizers in food processing applications. Specifically, significant reductions of biofilms of the pathogens Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes are observed, as are reductions in initial adhesion. Enzymes have the added benefits of being green, sustainable alternatives to chemical sanitizers, as well as having a minimal impact on food properties, in contrast to many alternative antimicrobial options such as bleach that aim to minimize food safety risks.

Escherichia coli O157:H7 and Salmonella Typhimurium adhesion to spinach leaf surfaces: Sensitivity to water chemistry and nutrient availability.

This study investigated the effects of solution chemistry and growth conditions on bacterial deposition on spinach leaf surfaces using a parallel plate flow cell. Two food safety pathogens of concern and two non-pathogen bacterial surrogates (environmental E. coli isolates) were grown in ideal (LB media) and nutrient-restricted (M9 media) conditions. Bacterial attachment was quantified as mass transfer rate coefficients for cells suspended in 10 mM KCl, CaCl2 and artificial groundwater, and cell and leaf surfaces were extensively characterized (zeta potential, hydrophobicity, extracellular polymer (EPS) composition). Between the pathogens, E. coli O157:H7 attachment was greater than that of Salmonella Typhimurium, attributed to measurable variability in cell surface charge and hydrophobicity. When grown in M9 media, both pathogens were significantly more adhesive to spinach surfaces (p < 0.01) than when grown in LB media. Surrogates did not follow this trend and showed minimal changes in adhesion kinetics and surface properties between growth conditions. EPS sugar/protein ratios were reduced in some of the highest attachment scenarios, suggesting that changes in EPS composition in favor of proteins may play a role. These results show the importance of growth conditions and solution complexities in understanding mechanisms of aqueous bacterial adhesion to food surfaces.

Comparison of filtration mechanisms of food and industrial grade TiO2 nanoparticles.

The removal of food and industrial grade titanium dioxide (TiO2) particles through drinking water filtration was assessed via direct visualization of an in situ 2-D micromodel. The goal of this research was to determine whether variances in surface composition, aggregate size, and ionic strength result in different transport and deposition processes in porous media. Food and industrial grade TiO2 particles were characterized by measuring their hydrodynamic diameter, zeta potential, and zero point of charge before introduction into the 2-D micromodel. The removal efficiency as a function of position on the collector surface was calculated from direct visualization measurements. Notably, food grade TiO2 had a lower removal efficiency when compared with industrial grade. The difference in removal efficiency between the two particle types could be attributed to the higher stability (as indicated by the larger zeta potential values) of the food grade particles, which lead to a reduced aggregate size when compared to the industrial grade particles. This removal efficiency trend was most pronounced in the rear stagnation point, due to the high contribution of hydrodynamic forces at that point. It could be inferred from the results presented herein that particle removal strategies should be based on particle aggregate size and surface charge. Graphical abstract ᅟ.

Influence of septic system wastewater treatment on titanium dioxide nanoparticle subsurface transport mechanisms.

Engineered nanomaterials (ENMs) are commonly incorporated into food and consumer applications to enhance a specific product aspect (i.e., optical properties). Life cycle analyses revealed ENMs can be released from products during usage and reach wastewater treatment plants (WWTPs), with titanium dioxide (TiO2) accounting for a large fraction. As such, food grade (FG) TiO2, a more common form of TiO2 in wastewater, was used in this study. Nanomaterials in WWTPs have been well characterized, although the problematic septic system has been neglected. Elution and bioaccumulation of TiO2 ENMs from WTTPs in downriver sediments and microorganisms has been observed; however, little is known about mechanisms governing the elution of FG TiO2 from the septic drainage system. This study characterized the transport behavior and mechanisms of FG TiO2 particles in porous media conditions after septic waste treatment. FG and industrial grade (IG) TiO2 (more commonly studied) were introduced to septic tank effluent and low-ionic strength electrolyte solutions prior to column transport experiments. Results indicate that FG TiO2 aggregate size (200-400 nm) remained consistent across solutions. Additionally, elution of FG and IG TiO2 was greatest in septic effluent at the higher nanoparticle concentration (100 ppm). FG TiO2 was well retained at the low (2 ppm) concentration in septic effluent, suggesting that particles that escape the septic system may still be retained in drainage field before reaching the groundwater system, although eluted particles are highly stabilized. Findings provide valuable insight into the significance of the solution environment at mediating differences observed between uniquely engineered nanomaterials. Graphical abstract.

Efficacy of post-harvest rinsing and bleach disinfection of E. coli O157:H7 on spinach leaf surfaces.

Attachment and detachment kinetics of Escherichia coli O157:H7 from baby spinach leaf epicuticle layers were investigated using a parallel plate flow chamber. Mass transfer rate coefficients were used to determine the impact of water chemistry and common bleach disinfection rinses on the removal and inactivation of the pathogen. Attachment mass transfer rate coefficients generally increased with ionic strength. Detachment mass transfer rate coefficients were nearly the same in KCl and AGW rinses; however, the detachment phase lasted longer in KCl than AGW (18 ± 4 min and 4 ± 2 min, respectively), indicating that the ions present during attachment play a significant role in the cells' ability to remain attached. Specifically, increasing bleach rinse concentration by two orders of magnitude was found to increase the detachment mass transfer rate coefficient by 20 times (from 5.7 ± 0.7 × 10-11 m/s to 112.1 ± 26.8 × 10-11 m/s for 10 ppb and 1000 ppb, respectively), and up to 88 ± 4% of attached cells remained alive. The spinach leaf texture was incorporated within a COMSOL model of disinfectant concentration gradients, which revealed nearly 15% of the leaf surface is exposed to almost 1000 times lower concentration than the bulk rinse solution.

O antigen modulates insect vector acquisition of the bacterial plant pathogen Xylella fastidiosa.

Hemipteran insect vectors transmit the majority of plant pathogens. Acquisition of pathogenic bacteria by these piercing/sucking insects requires intimate associations between the bacterial cells and insect surfaces. Lipopolysaccharide (LPS) is the predominant macromolecule displayed on the cell surface of Gram-negative bacteria and thus mediates bacterial interactions with the environment and potential hosts. We hypothesized that bacterial cell surface properties mediated by LPS would be important in modulating vector-pathogen interactions required for acquisition of the bacterial plant pathogen Xylella fastidiosa, the causative agent of Pierce's disease of grapevines. Utilizing a mutant that produces truncated O antigen (the terminal portion of the LPS molecule), we present results that link this LPS structural alteration to a significant decrease in the attachment of X. fastidiosa to blue-green sharpshooter foreguts. Scanning electron microscopy confirmed that this defect in initial attachment compromised subsequent biofilm formation within vector foreguts, thus impairing pathogen acquisition. We also establish a relationship between O antigen truncation and significant changes in the physiochemical properties of the cell, which in turn affect the dynamics of X. fastidiosa adhesion to the vector foregut. Lastly, we couple measurements of the physiochemical properties of the cell with hydrodynamic fluid shear rates to produce a Comsol model that predicts primary areas of bacterial colonization within blue-green sharpshooter foreguts, and we present experimental data that support the model. These results demonstrate that, in addition to reported protein adhesin-ligand interactions, O antigen is crucial for vector-pathogen interactions, specifically in the acquisition of this destructive agricultural pathogen.

D-amino acids inhibit initial bacterial adhesion: thermodynamic evidence.

Bacterial biofilms are structured communities of cells enclosed in a self-produced hydrated polymeric matrix that can adhere to inert or living surfaces. D-Amino acids were previously identified as self-produced compounds that mediate biofilm disassembly by causing the release of the protein component of the polymeric matrix. However, whether exogenous D-amino acids could inhibit initial bacterial adhesion is still unknown. Here, the effect of the exogenous amino acid D-tyrosine on initial bacterial adhesion was determined by combined use of chemical analysis, force spectroscopic measurement, and theoretical predictions. The surface thermodynamic theory demonstrated that the total interaction energy increased with more D-tyrosine, and the contribution of Lewis acid-base interactions relative to the change in the total interaction energy was much greater than the overall nonspecific interactions. Finally, atomic force microscopy analysis implied that the hydrogen bond numbers and adhesion forces decreased with the increase in D-tyrosine concentrations. D-Tyrosine contributed to the repulsive nature of the cell and ultimately led to the inhibition of bacterial adhesion. This study provides a new way to regulate biofilm formation by manipulating the contents of D-amino acids in natural or engineered systems.

Effect of hydration repulsion on nanoparticle agglomeration evaluated via a constant number Monte-Carlo simulation.

The effect of hydration repulsion on the agglomeration of nanoparticles in aqueous suspensions was investigated via the description of agglomeration by the Smoluchowski coagulation equation using constant number Monte-Carlo simulation making use of the classical DLVO theory extended to include the hydration repulsion energy. Evaluation of experimental DLS measurements for TiO2, CeO2, SiO2, and α-Fe2O3 (hematite) at high IS (up to 900 mM) or low |ζ-potential| (≥1.35 mV) demonstrated that hydration repulsion energy can be above electrostatic repulsion energy such that the increased overall repulsion energy can significantly lower the agglomerate diameter relative to the classical DLVO prediction. While the classical DLVO theory, which is reasonably applicable for agglomeration of NPs of high |ζ-potential| (∼>35 mV) in suspensions of low IS (∼<1 mM), it can overpredict agglomerate sizes by up to a factor of 5 at high IS or low |ζ-potential|. Given the potential important role of hydration repulsion over a range of relevant conditions, there is merit in quantifying this repulsion energy over a wide range of conditions as part of overall characterization of NP suspensions. Such information would be of relevance to improved understanding of NP agglomeration in aqueous suspensions and its correlation with NP physicochemical and solution properties.

Microbial Attachment Inhibition through Low-Voltage Electrochemical Reactions on Electrically Conducting Membranes.

Bacterial biofilm formation on membrane surfaces remains a serious challenge in water treatment systems. The impact of low voltages on microbial attachment to electrically conducting ultrafiltration membranes was investigated using a direct observation cross-flow membrane system mounted on a fluorescence microscope. Escherichia coli and microparticle deposition and detachment rates were measured as a function of the applied electrical potential to the membrane surface. Selecting bacteria and particles with low surface charge minimized electrostatic interactions between the bacteria and charged membrane surface. Application of an electrical potential had a significant impact on the detachment of live bacteria in comparison to dead bacteria and particles. Image analysis indicated that when a potential of 1.5 V was applied to the membrane/counter electrode pair, the percent of dead bacteria was 32±2.1 and 67±3.6% when the membrane was used as a cathode or anode, respectively, while at a potential of 1 V, 92±2.4% were alive. The application of low electrical potentials resulted in the production of low (µM) concentrations of hydrogen peroxide (HP) through the electroreduction of oxygen. The electrochemically produced HP reduced microbial cell viability and increased cellular permeability. Exposure to low concentrations of electrochemically produced HP on the membrane surface prevents bacterial attachment, thus ensuring biofilm-free conditions during membrane filtration operations.

Soybean susceptibility to manufactured nanomaterials with evidence for food quality and soil fertility interruption.

Based on previously published hydroponic plant, planktonic bacterial, and soil microbial community research, manufactured nanomaterial (MNM) environmental buildup could profoundly alter soil-based food crop quality and yield. However, thus far, no single study has at once examined the full implications, as no studies have involved growing plants to full maturity in MNM-contaminated field soil. We have done so for soybean, a major global commodity crop, using farm soil amended with two high-production metal oxide MNMs (nano-CeO(2) and -ZnO). The results provide a clear, but unfortunate, view of what could arise over the long term: (i) for nano-ZnO, component metal was taken up and distributed throughout edible plant tissues; (ii) for nano-CeO(2), plant growth and yield diminished, but also (iii) nitrogen fixation--a major ecosystem service of leguminous crops--was shut down at high nano-CeO(2) concentration. Juxtaposed against widespread land application of wastewater treatment biosolids to food crops, these findings forewarn of agriculturally associated human and environmental risks from the accelerating use of MNMs.

Fate and Transport of Molybdenum Disulfide Nanomaterials in Sand Columns.

Research and development of two-dimensional transition metal dichalcogenides (TMDC) (e.g., molybdenum disulfide [MoS2]) in electronic, optical, and catalytic applications has been growing rapidly. However, there is little known regarding the behavior of these particles once released into aquatic environments. Therefore, an in-depth study regarding the fate and transport of two popular types of MoS2 nanomaterials, lithiated (MoS2-Li) and Pluronic PF-87 dispersed (MoS2-PL), was conducted in saturated porous media (quartz sand) to identify which form would be least mobile in aquatic environments. The electrokinetic properties and hydrodynamic diameters of MoS2 as a function of ionic strength and pH were determined using a zeta potential analyzer and dynamic light scattering techniques. Results suggest that the stability is significantly decreased beginning at 10 and 31.6 mM KCl, for MoS2-PL and MoS2-Li, respectively. Transport study results from breakthrough curves, column dissections, and release experiments suggest that MoS2-PL exhibits a greater affinity to be irreversibly bound to quartz surfaces as compared with the MoS2-Li at a similar ionic strength. Derjaguin-Landau-Verwey-Overbeek theory was used to help explain the unique interactions between the MoS2-PL and MoS2-Li surfaces between particles and with the quartz collectors. Overall, the results suggest that the fate and transport of MoS2 is dependent on the type of MoS2 that enters the environment, where MoS2-PL will be least mobile and more likely be deposited in porous media from pluronic-quartz interactions, whereas MoS2-Li will travel greater distances and have a greater tendency to be remobilized in sand columns.

Epithelial microvilli establish an electrostatic barrier to microbial adhesion.

Microvilli are membrane extensions on the apical surface of polarized epithelia, such as intestinal enterocytes and tubule and duct epithelia. One notable exception in mucosal epithelia is M cells, which are specialized for capturing luminal microbial particles; M cells display a unique apical membrane lacking microvilli. Based on studies of M cell uptake under different ionic conditions, we hypothesized that microvilli may augment the mucosal barrier by providing an increased surface charge density from the increased membrane surface and associated glycoproteins. Thus, electrostatic charges may repel microbes from epithelial cells bearing microvilli, while M cells are more susceptible to microbial adhesion. To test the role of microvilli in bacterial adhesion and uptake, we developed polarized intestinal epithelial cells with reduced microvilli ("microvillus-minus," or MVM) but retaining normal tight junctions. When tested for interactions with microbial particles in suspension, MVM cells showed greatly enhanced adhesion and uptake of particles compared to microvillus-positive cells. This preference showed a linear relationship to bacterial surface charge, suggesting that microvilli resist binding of microbes by using electrostatic repulsion. Moreover, this predicts that pathogen modification of electrostatic forces may contribute directly to virulence. Accordingly, the effacement effector protein Tir from enterohemorrhagic Escherichia coli O157:H7 expressed in epithelial cells induced a loss of microvilli with consequent enhanced microbial binding. These results provide a new context for microvillus function in the host-pathogen relationship, based on electrostatic interactions.

Interactions of glycosphingolipids and lipopolysaccharides with silica and polyamide surfaces: adsorption and viscoelastic properties.

Bacterial outer membrane components play a critical role in bacteria-surface interactions (adhesion and repulsion). Sphingomonas species (spp.) differ from other Gram-negative bacteria in that they lack lipopolysaccharides (LPSs) in their outer membrane. Instead, Sphingomonas spp. outer membrane consists of glycosphingolipids (GSLs). To delineate the properties of the outer membrane of Sphingomonas spp. and to explain the adhesion of these cells to surfaces, we employed a single-component-based approach of comparing GSL vesicles to LPS vesicles. This is the first study to report the formation of vesicles containing 100% GSL. Significant physicochemical differences between GSL and LPS vesicles are reported. Composition-dependent vesicle adherence to different surfaces using quartz crystal microbalance with dissipation monitoring (QCM-D) technology was observed, where higher GSL content resulted in higher mass accumulation on the sensor. Additionally, the presence of 10% GSL and above was found to promote the relative rigidity of the vesicle obtaining viscoelastic ratio of 30-70% higher than that of pure LPS vesicles.

Biofouling of reverse osmosis membranes: positively contributing factors of Sphingomonas.

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.

Soybean plants modify metal oxide nanoparticle effects on soil bacterial communities.

Engineered nanoparticles (ENPs) are entering agricultural soils through land application of nanocontaining biosolids and agrochemicals. The potential adverse effects of ENPs have been studied on food crops and soil bacterial communities separately; however, how ENPs will affect the interacting plant-soil system remains unknown. To address this, we assessed ENP effects on soil microbial communities in soybean-planted, versus unplanted, mesocosms exposed to different doses of nano-CeO2 (0-1.0 g kg(-1)) or nano-ZnO (0-0.5 g kg(-1)). Nano-CeO2 did not affect soil bacterial communities in unplanted soils, but 0.1 g kg(-1) nano-CeO2 altered soil bacterial communities in planted soils, indicating that plants interactively promote nano-CeO2 effects in soil, possibly due to belowground C shifts since plant growth was impacted. Nano-ZnO at 0.5 g kg(-1) significantly altered soil bacterial communities, increasing some (e.g., Rhizobium and Sphingomonas) but decreasing other (e.g., Ensifer, Rhodospirillaceae, Clostridium, and Azotobacter) operational taxonomic units (OTUs). Fewer OTUs decreased from nano-ZnO exposure in planted (41) versus unplanted (85) soils, suggesting that plants ameliorate nano-ZnO effects. Taken together, plants--potentially through their effects on belowground biogeochemistry--could either promote (i.e., for the 0.1 g kg(-1) nano-CeO2 treatment) or limit (i.e., for the 0.5 g kg(-1) nano-ZnO treatment) ENP effects on soil bacterial communities.

Transport of , , and microspheres in biochar-amended soils with different textures.

The incorporation of biochar into soils has been proposed as a means to sequester carbon from the atmosphere. An added environmental benefit is that biochar has been shown to increase soil retention of agrochemicals, and recent research has indicated that biochar may be effective in increasing soil retention of bacteria. In this study we investigate the transport behavior of O157:H7, serovar Typhimurium, and carboxylated polystyrene microspheres in water-saturated column experiments for two soils (fine sand and sandy loam) amended with 2% poultry litter or pine chip biochars pyrolyzed at 350 and 700°C. Adding poultry litter biochar pyrolyzed at 350°C did not improve soil retention of either bacteria in fine sand and even facilitated their transport in sandy loam. Addition of either biochar pyrolyzed at 700°C generally improved retention of bacteria in fine sand, with the pine chip biochars being more effective in limiting their transport. Results from the column studies and auxiliary batch studies suggest that changes in cell retention after biochar amendments were likely due to changes in bacterial attachment in the column and not to physical straining or changes in survivability. We also found that changes in bacterial hydrophobicity after biochar amendments were generally correlated with changes in bacterial retention. The influence of biochar amendment in increasing retention of both bacteria was generally more pronounced in fine sand and indicates that soil texture affects the transport behavior of bacteria through biochar-amended soils.

Removal of TiO2 Nanoparticles During Primary Water Treatment: Role of Coagulant Type, Dose, and Nanoparticle Concentration.

Nanomaterials from consumer products (i.e., paints, sunscreens, toothpastes, and food grade titanium dioxide [TiO2]) have the capacity to end up in groundwater and surface water, which is of concern because the effectiveness of removing them via traditional treatment is uncertain. Although aggregation and transport of nanomaterials have been investigated, studies on their removal from suspension are limited. Hence, this study involves the development of scaled-down jar tests to determine the mechanisms involved in the removal of a model metal oxide nanoparticle (NP), TiO2, in artificial groundwater (AGW), and artificial surface water (ASW) at the primary stages of treatment: coagulation, flocculation, and sedimentation. Total removal was quantified at the end of each treatment stage by spectroscopy. Three different coagulants-iron chloride (FeCl3), iron sulfate (FeSO4), and alum [Al2(SO4)3]-destabilized the TiO2 NPs in both source waters. Overall, greater than one-log removal was seen in groundwater for all coagulants at a constant dose of 50 mg/L and across the range of particle concentrations (10, 25, 50, and 100 mg/L). In surface water, greater than 90% removal was seen with FeSO4 and Al2(SO4)3, but less than 60% when using FeCl3. Additionally, removal was most effective at higher NP concentrations (50 and 100 mg/L) in AGW when compared with ASW. Zeta potential was measured and compared between AGW and ASW with the presence of all three coagulants at the same treatment stage times as in the removal studies. These electrokinetic trends confirm that the greatest total removal of NPs occurred when the magnitude of charge was smallest (<10 mV) and conversely, higher zeta potential values (>35 mV) measured were under conditions with poor removal (<90%). These results are anticipated to be of considerable interest to practitioners for the assessment of traditional treatment processes' capacity to remove nanomaterials prior to subsequent filtration and distribution to domestic water supplies.

Stability and Transport of Graphene Oxide Nanoparticles in Groundwater and Surface Water.

The effects of groundwater and surface water constituents (i.e., natural organic matter [NOM] and the presence of a complex assortment of ions) on graphene oxide nanoparticles (GONPs) were investigated to provide additional insight into the factors contributing to fate and the mechanisms involved in their transport in soil, groundwater, and surface water environments. The stability and transport of GONPs was investigated using dynamic light scattering, electrokinetic characterization, and packed bed column experiments. Stability results showed that the hydrodynamic diameter of the GONPs at a similar ionic strength (2.1±1.1 mM) was 10 times greater in groundwater environments compared with surface water and NaCl and MgCl2 suspensions. Transport results confirmed that in groundwater, GONPs are less stable and are more likely to be removed during transport in porous media. In surface water and MgCl2 and NaCl suspensions, the relative recovery was 94%±3% indicating that GONPs will be very mobile in surface waters. Additional experiments were carried out in monovalent (KCl) and divalent (CaCl2) salts across an environmentally relevant concentration range (0.1-10 mg/L) of NOM using Suwannee River humic acid. Overall, the transport and stability of GONPs was increased in the presence of NOM. This study confirms that planar "carbonaceous-oxide" materials follow traditional theory for stability and transport, both due to their response to ionic strength, valence, and NOM presence and is the first to look at GONP transport across a wide range of representative conditions found in surface and groundwater environments.

Linking microbial community structure to function in representative simulated systems.

Pathogenic bacteria are generally studied as a single strain under ideal growing conditions, although these conditions are not the norm in the environments in which pathogens typically proliferate. In this investigation, a representative microbial community along with Escherichia coli O157:H7, a model pathogen, was studied in three environments in which such a pathogen could be found: a human colon, a septic tank, and groundwater. Each of these systems was built in the lab in order to retain the physical/chemical and microbial complexity of the environments while maintaining control of the feed into the models. The microbial community in the colon was found to have a high percentage of bacteriodetes and firmicutes, while the septic tank and groundwater systems were composed mostly of proteobacteria. The introduction of E. coli O157:H7 into the simulated systems elicited a shift in the structures and phenotypic cell characteristics of the microbial communities. The fate and transport of the microbial community with E. coli O157:H7 were found to be significantly different from those of E. coli O157:H7 studied as a single isolate, suggesting that the behavior of the organism in the environment was different from that previously conceived. The findings in this study clearly suggest that to gain insight into the fate of pathogens, cells should be grown and analyzed under conditions simulating those of the environment in which the pathogens are present.

Deposition and survival of Escherichia coli O157:H7 on clay minerals in a parallel plate flow system.

Understanding bacterial pathogens deposition and survival processes in the soil-groundwater system is crucial to protect public health from soilborne and waterborne diseases. However, mechanisms of bacterial pathogen-clay interactions are not well studied, particularly in dynamic systems. Also, little is known about the viability of bacterial pathogens when attached to clays. In this study, a parallel plate flow system was used to determine the deposition kinetics and survival of Escherichia coli O157:H7 on montmorillonite, kaolinite, and goethite over a wide range of ionic strengths (IS) (0.1-100 mM KCl). E. coli O157:H7 deposition on the positively charged goethite is greater than that on the negatively charged kaolinite and montmorillonite. Although the zeta potential of kaolinite was more negative than that of montmorillonite, kaolinite showed a greater deposition for E. coli O157:H7 than montmorillonite, which is attributed to the chemical heterogeneity of clay minerals. Overall, increasing IS resulted in an increase of E. coli O157:H7 deposition on montmorillonite and kaolinite, and a decrease on goethite. Interaction energy calculations suggest that E. coli O157:H7 deposition on clays was largely governed by DLVO (Derjaguin-Landau-Verwey-Overbeek) forces. The loss of bacterial membrane integrity was investigated as a function of time using the Live/Dead BacLight viability assay. During the examined period of 6 h, E. coli O157:H7 retained its viability in suspension and when attached to montmorillonite and kaolinite; however, interaction with the goethite was detrimental. The information obtained in this study is of fundamental significance for the understanding of the fate of bacterial pathogens in soil environments.