Antifouling Properties of a Self-Assembling Glutamic Acid-Lysine Zwitterionic Polymer Surface Coating.

There is a need for the development of antifouling materials to resist adsorption of biomacromolecules. Here we describe the preparation of a novel zwitterionic block copolymer with the potential to prevent or delay the formation of microbial biofilms. The block copolymer comprised a zwitterionic (hydrophilic) section of alternating glutamic acid (negatively charged) and lysine (positively charged) units and a hydrophobic polystyrene section. Cryo-TEM and dynamic-light-scattering (DLS) results showed that, on average, the block copolymer self-assembled into 7-nm-diameter micelles in aqueous solutions (0 to 100 mM NaCl, pH 6). Quartz crystal microbalance with dissipation monitoring (QCM-D), atomic force microscopy (AFM), and contact angle measurements demonstrated that the block copolymer self-assembled into a brush-like monolayer on polystyrene surfaces. The brush-like monolayer produced from a 100 mg/L block copolymer solution exhibited an average distance, d, of approximately 4-8 nm between each block copolymer molecule (center to center). Once the brush-like monolayer self-assembled, it reduced EPS adsorption onto the polystyrene surface by ∼70% (mass), reduced the rate of bacterial attachment by >80%, and inhibited the development of thick biofilms. QCM-D results revealed that the EPS molecules penetrate between the chains of the brush and adsorb onto the polystyrene surface. Additionally, AFM analyses showed that the brush-like monolayer prevents the adhesion of large (> d) hydrophilic colloids onto the surface via hydration repulsion; however, molecules or colloids small enough to fit between the brush polymers (< d) were able to be adsorbed onto the surface via van der Waals interactions. Overall, we found that the penetration of extracellular organelles, as well as biopolymers through the brush, is critical for the failure of the antifouling coating, and likely could be prevented through tuning of the brush density. Stability and biofilm development testing on multiple surfaces (polypropylene, glass, and stainless steel) support practical applications of this novel block copolymer.

Disruptions in loading and aeration impact effluent chlorine demand during biological greywater recycling.

Greywater recycling systems designed for high-quality applications, such as hand washing, must deliver microbially safe and aesthetically acceptable water under the challenging operating conditions present where such systems are needed most urgently. As chlorination is the most popular strategy for reducing bacterial concentrations in greywater, understanding chlorination in the context of disruptive and challenging operation is essential to designing robust treatment. In this study, we have examined how disruptions through overall increased loading, interrupted aeration and increased ammonia loading have impacted the chlorine demand of the water produced by a greywater recycling system. We also presented concentrations of significant chemicals that contributed to this chlorine demand. The results indicate that a 1 d period with 8 times (8x) the normal design loading produced a peak chlorine demand of 0.74 mg Cl2/L, which is approximately double the baseline value. While this chlorine demand can be overcome by adding more chlorine, tests involving disruptions in aeration or feeding additional ammonia into the bioreactor produced much greater increases (>30x). The risks of increased chlorine demand on microbial safety can be overcome by limiting ammonia inputs to the system, providing backup systems to ensure sufficient aeration, or through additional anti-bacterial measures that do not depend on maintaining residual chlorine.

Linking transformations of organic carbon to post-treatment performance in a biological water recycling system.

Ozone, electrolysis and granular activated carbon (GAC) were examined as potential post-treatments to follow a household-scale biologically activated membrane bioreactor (BAMBi), treating a wash water containing trace urine and feces contamination. Each post-treatment was evaluated for abilities and reaction preferences to remove or transform dissolved organic carbon (DOC), chemical structures that contribute color, and assimilable organic carbon (AOC), which can support bacterial regrowth. Batch treatment with each technology demonstrated an ability to remove ≥95% DOC. Ozone demonstrated a reaction selectivity through increased reaction rates with larger compounds and color-contributing compounds. Electrolysis and GAC demonstrated generally less-selective reactivity. Adding post-treatments to full-scale systems reduced DOC (55-91%), AOC (34-62%), and color (75-98%), without significant reaction selectivity. These reductions in DOC and AOC were not linked to reduction of bacterial concentrations in treated water. Reductions in bacterial concentrations were observed with ozone and electrolysis, but this is credited to oxidation chemicals produced in these systems and not the removal or transformations of organic materials.

Comparing the anti-bacterial performance of chlorination and electrolysis post-treatments in a hand washing water recycling system.

Innovative solutions are necessary to enable the decentralized recycling of greywater for applications requiring high-quality water, such as hand washing. While physical barriers such as ultrafiltration membranes effectively prevent the passage of bacteria, and chemical and biological treatments can effectively reduce the carbon content of the treated water, there exists a knowledge gap regarding the application of anti-bacterial strategies to prevent the growth of harmful bacteria following treatment. In this study, the effluent water from a household-scale greywater treatment system was fed to seven parallel experimental post-treatment tanks: three receiving direct chlorination with free chlorine residuals of 0.2, 1 or 5 mg Cl2/L, three with chlorine produced through electrolysis at the same residual concentrations, and one control with no chlorine added. For increasing concentrations of direct chlorination, the median total cell count (TCC) values were 9 × 104, 2.9 × 104 and 1.8 × 103 cells/mL, respectively. Electrolysis treatment produced very similar TCC concentrations, 8.8 × 104, 1.1 × 104 and 2.3 × 103 cells/mL. The TCC concentrations were lower than the concentration of the water entering each tank (∼3 × 105 cells/mL). Intact cell count (ICC) measurements indicated that the viable cell concentrations, were less than 10% of the TCC values. Though electrolysis treatment can produce powerful oxidants, such as hydroxyl radical, there was no evidence that electrolysis in this system provided additional benefits beyond chlorine production for control of total or intact cell counts. Oxidation of bacteria by chlorine was the dominant anti-bacterial mechanism in our system. Monitoring of dissolved organic carbon (DOC) and assimilable organic carbon (AOC) did not suggest that carbon-limitation significantly impacted cell counts when chlorination or electrolysis treatment was applied. This work demonstrates that either direct chlorination or electrolysis treatment are able to reduce bacteria concentrations over long-term operation of a hand washing water treatment system. We recommend selecting chlorine residual targets such that a chlorine residual is maintained during periods of challenging operating conditions. We observed that a target residual of 1 mg Cl2/L, in our system, maintained the TCC below the concentration found in Zurich drinking water.

Chemical composition, nutrient-balancing and biological treatment of hand washing greywater.

On-site biological hand washing water treatment can improve global access to safe hand washing water, but requires a thorough understanding of the chemical composition of the water to be treated, and an effective treatment strategy. This study first presents a detailed characterization of the individual inputs to hand washing water. We demonstrate (i) that soap is likely the most significant input in hand washing water, representing ∼90% of mass loading, and (ii) that inputs to hand washing water have low concentrations of biologically-essential macro- and micro-nutrients (nitrogen, phosphorus, potassium, copper, zinc, molybdenum and cobalt) with respect to carbon, which may impair biological carbon removal. This study next formulates a recipe that recreates a representative composition of hand washing water and develops a procedure to identify and supplement nutrients in which this recipe is estimated to be deficient. Batch testing of the nutrient-supplemented hand washing water with an inoculum of planktonic bacteria demonstrated improved assimilable organic carbon removal (99% vs. 86% removal) and produced lower final dissolved organic carbon concentrations (1.7 mgC/L vs. 3.5 mgC/L) compared to realistic (nutrient-deficient) washing water. Supplementing nutrients did promote cell growth (50x higher final total cell count). Full-scale testing in a biologically activated membrane bioreactor (BAMBi) system treating 75 L/day of nutrient-supplemented hand washing water showed that long-term operation (100 days) can deliver effective carbon removal (95%) without detrimental fouling or other disruptions caused by cell growth. This work demonstrates that biological treatment in a BAMBi system, operated with appropriate nutrient-balancing offers an effective solution for decentralized treatment of light greywater.

Controlling Bacterial Pathogens in Water for Reuse: Treatment Technologies for Water Recirculation in the Blue Diversion Autarky Toilet.

The Blue Diversion AUTARKY Toilet is a urine-diverting toilet with on-site treatment. The toilet is being developed to provide a safe and affordable sanitation technology for people who lack access to sewer-based sanitation. Water used for personal hygiene, hand washing, and flushing to rinse urine- and feces-collection bowls is treated, stored, and recycled for reuse to reduce reliance on external water supplies. The system provides an opportunity to investigate hygiene of water for reuse following treatment. Treatment in the toilet includes a Biologically Activated Membrane Bioreactor (BAMBi) followed by a secondary treatment technology. To identify effective secondary treatment, three options, including granular activated carbon (GAC) only, GAC+chlorine (sodium hypochlorite), and GAC+electrolysis are considered based on the bacterial inactivation and growth inhibition efficiency. Four different hygiene-relevant bacteria are tested: Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, and Salmonella typhimurium. Our evaluation demonstrates that-despite treatment of water with the BAMBi-E. coli, P aeruginosa, and S. typhimurium have the potential to grow during storage in the absence of microbial competition. Including the indigenous microbial community influences bacterial growth in different ways: E. coli growth decreases but P. aeruginosa growth increases relative to no competition. The addition of the secondary treatment options considerably improves water quality. A column of GAC after the BAMBi reduces E. coli growth potential by 2 log10, likely due to the reduction of carbon sources. Additional treatments including chlorination and electrolysis provide further safety margins, with more than 5 log-10 inactivation of E. coli. However, reactivation and/or regrowth of E. coli and P. aeruginosa occurs under in the absence of residual disinfectant. Treatment including the BAMBi, GAC, and electrolysis appear to be promising technologies to control bacterial growth during storage in water intended for reuse.

Cellulose effects on morphology and elasticity of Vibrio fischeri biofilms.

Cellulose effects on Vibrio fischeri biofilm morphology were tested for the wild-type and two of its isogenic mutants that either exhibit increased cellulose production or do not produce cellulose at all. Confocal laser scanning microscopy imaging of each biofilm revealed that total sessile volume increases with cellulose expression, but the size of colonies formed with cellulose was smaller, creating a more diffuse biofilm. These morphological differences were not attributed to variations in bacterial deposition, extracellular polymeric substances affinity to the surface or bacterial growth. A positive correlation was found between cellulose expression, Young's (elastic) modulus of the biofilm analyzed with atomic force microscope and shear modulus of the related extracellular polymeric substances layers analyzed with quartz crystal microbalance with dissipation monitoring. Cellulose production also correlated positively with concentrations of extracellular DNA. A significant negative correlation was observed between cellulose expression and rates of diffusion through the extracellular polymeric substances. The difference observed in biofilm morphology is suggested as a combined result of cellulose and likely extracellular DNA (i) increasing biofilm Young's modulus, making shear removal more difficult, and (ii) decreased diffusion rate of nutrients and wastes into and out of the biofilm, which effectively limits colony size.

Net energy production associated with pathogen inactivation during mesophilic and thermophilic anaerobic digestion of sewage sludge.

The potential for anaerobic digester energy production must be balanced with the sustainability of reusing the resultant biosolids for land application. Mesophilic, thermophilic, temperature-phased, and high temperature (60 or 70 °C) batch pre-treatment digester configurations have been systematically evaluated for net energy production and pathogen inactivation potential. Energy input requirements and net energy production were modeled for each digester scheme. First-order inactivation rate coefficients for Escherichia coli, Enterococcus faecalis and bacteriophage MS-2 were measured at each digester temperature and full-scale pathogen inactivation performance was estimated for each indicator organism and each digester configuration. Inactivation rates were found to increase dramatically at temperatures above 55 °C. Modeling full-scale performance using retention times based on U.S. EPA time and temperature constraints predicts a 1-2 log inactivation in mesophilic treatment, and a 2-5 log inactivation in 50-55 °C thermophilic and temperature-phased treatments. Incorporating a 60 or 70 °C batch pre-treatment phase resulted in dramatically higher potency, achieving MS-2 inactivation of 14 and 16 logs respectively, and complete inactivation (over 100 log reduction) of E. coli and E. faecalis. For temperatures less than 70 °C, viability staining of thermally-treated E. coli showed significantly reduced inactivation relative to standard culture enumeration. Due to shorter residence times in thermophilic reactors, the net energy production for all digesters was similar (less than 20% difference) with the 60 or 70 °C batch treatment configurations producing the most net energy and the mesophilic treatment producing the least. Incorporating a 60 or 70 °C pre-treatment phase can dramatically increase pathogen inactivation performance without decreasing net energy capture from anaerobic digestion. Energy consumption is not a significant barrier against improving the pathogen quality of biosolids.

Impact of higher alginate expression on deposition of Pseudomonas aeruginosa in radial stagnation point flow and reverse osmosis systems.

Extracellular polymeric substances (EPS) have major impact on biofouling of reverse osmosis (RO) membranes. On one hand, EPS can reduce membrane permeability and on the other, EPS production by the primary colonizers may influence their deposition and attachment rate and subsequently affect the biofouling propensity of the membrane. The role of bacterial exopolysaccharides in bacterial deposition followed by the biofouling potential of an RO membrane was evaluated using an alginate overproducing (mucoid) Pseudomonas aeruginosa. The mucoid P. aeruginosa PAOmucA22 was compared with its isogenic nonmucoid prototypic parent PAO1 microscopically in a radial stagnation point flow (RSPF) system for their bacterial deposition characteristics. Then, biofouling potential of PAO1 and PAOmucA22 was determined in a crossflow rectangular plate-and-frame membrane cell, in which the strains were cultivated on a thin-film composite, polyamide, flat RO membrane coupon (LFC-1) under laminar flow conditions. In the RSPF system, the observed deposition rate of the mucoid strain was between 5- and 10-fold lower than of the wild type using either synthetic wastewater medium (with ionic strength of 14.7 mM and pH 7.4) or 15 mM KCl solution (pH of 6.2). The slower deposition rate of the mucoid strain is explained by 5- to 25-fold increased hydrophilicity of the mucoid strain as compared to the isogenic wild type, PAO1. Corroborating with these results, a significant delay in the onset of biofouling of the RO membrane was observed when the mucoid strain was used as the membrane colonizer, in which the observed time for the induced permeate flux decline was delayed (ca. 2-fold). In conclusion, the lower initial cell attachment of the mucoid strain decelerated biofouling of the RO membrane. Bacterial deposition and attachment is a critical step in biofilm formation and governed by intimate interactions between outer membrane proteins of the bacteria and the surface. Shielding these interactions by a hydrated and hydrophilic alginate capsule is shown to dramatically lessen the biofouling potential of the membrane colonizers.