Real-time biomonitoring of oxygen uptake rate and biochemical oxygen demand using a novel optical biogas respirometric system.

In response to the ever-increasing need for monitoring-based process control of wastewater treatment plants, an online applicable respirometer shows great promise for real-time measurement of oxygen uptake rate (OUR) and biochemical oxygen demand (BOD) measurements as a surrogate of the biodegradability of wastewater. Here, we have developed a photosensor-assisted real-time respirometric system equipped with bubble counting sensors for accurate measurement of microbial oxygen consumption in a bottle. This system can measure OUR and BOD in a bottle equipped with a tube containing NaOH solution to absorb carbon dioxide and supplied with continuous atmospheric oxygen to the bottle, which reliably supplies non-limiting dissolved oxygen (DO) for aerobic biodegradation even at high organic loads. These technical improvements allow a sensitive and rapid analytical tool offering real-time profiles of oxygen uptake rate as well as BOD measurements with an extended measurable range (0-420 mg O2/L), enabling significant reduction or elimination of dilution steps. The respirometric system was used to elucidate the biodegradable kinetics of domestic and swine wastewaters as a function of the type and concentration of organic matters, depending on source characteristics including rapidly or slowly oxidizable organic substances by bacteria. Compared with conventional and manometric BOD methods, our method is reliable and accurate.

Tunable Ion Sieving of Graphene Membranes through the Control of Nitrogen-Bonding Configuration.

Graphene-oxide (GO) membranes with notable ionic-sieving properties have attracted significant attention for many applications. However, the swelling and unstable nanostructure of GO laminates in water results in enlarged interlayer spacing and a low permeation cut-off, limiting their applicability for water purification and desalination. Herein, we propose novel nitrogen-doped graphene (NG) membranes for use in tunable ion sieving that are made via facile fabrication by a time-dependent N-doping technique. Doping reaction time associated variation in atomic content and bonding configurations strongly contributed to the nanostructure of NG laminates by yielding narrower interlayer spacing and a more-polarized surface than GO. These nanostructural features subsequently allowed ion transport through the combined mechanisms of size exclusion and electrostatic interaction. The stacked NG membranes provided size-dependent permeability for hydrated ions and improved ion selectivity by 1-3 orders of magnitude in comparison to that of a GO membrane. For ions small enough to move through the interlayer spacing, the ion permeation is determined by electrostatic properties of NG membranes with the type of N configuration, especially polarized pyridinic N. Due to these properties, the NG membrane functioned as an unconventionally selective graphene-based membrane with better ion sieving for water purification.

Near Real-Time Detection of E. coli in Reclaimed Water.

Advanced treatment of reclaimed water prior to potable reuse normally results in the inactivation of bacterial populations, however, incremental treatment failure can result in bacteria, including pathogens, remaining viable. Therefore, potential microorganisms need to be detected in real-time to preclude potential adverse human health effects. Real-time detection of microbes presents unique problems which are dependent on the water quality of the test water, including parameters such as particulate content and turbidity, and natural organic matter content. In addition, microbes are unusual in that: (i) viability and culturability are not always synonymous; (ii) viability in water can be reduced by osmotic stress; and (iii) bacteria can invoke repair mechanisms in response to UV disinfection resulting in regrowth of bacterial populations. All these issues related to bacteria affect the efficacy of real-time detection for bacteria. Here we evaluate three different sensors suitable for specific water qualities. The sensor A is an on-line, real-time sensor that allows for the continuous monitoring of particulates (including microbial contaminants) using multi-angle-light scattering (MALS) technology. The sensor B is a microbial detection system that uses optical technique, Mie light scattering, for particle sizing and fluorescence emission for viable bacteria detection. The last sensor C was based on adenosine triphosphate (ATP) production. E. coli was used a model organism and out of all tested sensors, we found the sensor C to be the most accurate. It has a great potential as a surrogate parameter for microbial loads in test waters and be useful for process control in treatment trains.

Physicochemical interactions between rhamnolipids and Pseudomonas aeruginosa biofilm layers.

This study investigated the physicochemical interactions between a rhamnolipid biosurfactant and a biofilm layer. A concentration of 300 µg mL(-1) of rhamnolipids, which is around the critical micelle concentration value (240 µg mL(-1)), showed great potential for reducing biofilm. The surface free energy between the rhamnolipids and biofilm layer decreased, as did the negative surface charge, due to the removal of negatively charged humic-like, protein-like, and fulvic acid-like substances. The carbohydrate and protein concentrations composed of extracellular polymeric substances decreased by 31.6% and 79.6%, respectively, at a rhamnolipid concentration of 300 µg mL(-1). In particular, rhamnolipids can interact with proteins, leading to a reduction of the N source and amide groups on the membrane. For carbohydrates, the component ratio of glucosamine was decreased, but the levels of glucose and mannose that form the majority of the carbohydrates remained unchanged. To our knowledge, the present study is the first attempt at studying the interactions of the two phases of rhamnolipids and the biofilm layer, and as such is expected to clarify the mechanism by which rhamnolipids lead to a reduction in biofilm.

Use of rhamnolipid biosurfactant for membrane biofouling prevention and cleaning.

Rhamnolipids were evaluated as biofouling reducing agents in this study. The permeability of the bacterial outer membrane was increased by rhamnolipids while the growth rate of Pseudomonas aeruginosa was not affected. The surface hydrophobicity was increased through the release of lipopolysaccharides and extracellular polymeric substances from the outer cell membrane. Rhamnolipids were evaluated as agents for the prevention and cleaning of biofilms. A high degree of biofilm detachment was observed when the rhamnolipids were used as a cleaning agent. In addition, effective biofilm reduction occurred when rhamnolipids were applied to various species of Gram-negative bacteria isolated from seawater samples. Biofilm reduction using rhamnolipids was comparable to commercially available surfactants. In addition, 20% of the water flux was increased after rhamnolipid treatment (300 µg ml(-1), 6 h exposure time) in a dead-end filtration system. Rhamnolipids appear to have promise as biological agents for reducing membrane biofouling.

Comparative pyrosequencing analysis of bacterial community change in biofilm formed on seawater reverse osmosis membrane.

The change in bacterial community structure induced by bacterial competition and succession was investigated during seawater reverse osmosis (SWRO) in order to elucidate a possible link between the bacterial consortium on SWRO membranes and biofouling. To date, there has been no definitive characterization of the microbial diversity in SWRO in terms of distinguishing time-dependent changes in the richness or abundance of bacterial species. For bacterial succession within biofilms on the membrane surface, SWRO using a cross-flow filtration membrane test unit was operated for 5 and 100h, respectively. As results of the pyrosequencing analysis, bacterial communities differed considerably among seawater and the 5 and 100 h samples. From a total of 33,876 pyrosequences (using a 95% sequence similarity), there were less than 1% of shared species, confirming the influence of the operational time factor and lack of similarity of these communities. During SWRO operation, the abundance of Pseudomonas stutzeri BBSPN3 (GU594474) belonging to gamma-Proteobacteria suggest that biofouling of SWRO membrane might be driven by the dominant influence of a specific species. In addition, among the bacterial competition of five bacterial species (Pseudomonas aeruginosa, Bacillus sp., Rhodobacter sp., Flavobacterium sp., and Mycobacterium sp.) competing for bacterial colonization on the SWRO membrane surfaces, it was exhibited that Bacillus sp. was the most dominant. The dominant influences ofPseudomonas sp. and Bacillus sp. on biofouling during actual SWRO is decisive depending on higher removal efficiency of the seawater pretreatment.

Bead-based competitive fluorescence immunoassay for sensitive and rapid diagnosis of cyanotoxin risk in drinking water.

Due to the increased occurrence of cyanobacterial blooms and their toxins in drinking water sources, effective management based on a sensitive and rapid analytical method is in high demand for security of safe water sources and environmental human health. Here, a competitive fluorescence immunoassay of microcystin-LR (MCYST-LR) is developed in an attempt to improve the sensitivity, analysis time, and ease-of-manipulation of analysis. To serve this aim, a bead-based suspension assay was introduced based on two major sensing elements: an antibody-conjugated quantum dot (QD) detection probe and an antigen-immobilized magnetic bead (MB) competitor. The assay was composed of three steps: the competitive immunological reaction of QD detection probes against analytes and MB competitors, magnetic separation and washing, and the optical signal generation of QDs. The fluorescence intensity was found to be inversely proportional to the MCYST-LR concentration. Under optimized conditions, the proposed assay performed well for the identification and quantitative analysis of MCYST-LR (within 30 min in the range of 0.42-25 µg/L, with a limit of detection of 0.03 µg/L). It is thus expected that this enhanced assay can contribute both to the sensitive and rapid diagnosis of cyanotoxin risk in drinking water and effective management procedures.

Flow cytometric detection of Bacillus spoOA gene in biofilm using quantum dot labeling.

Quantum dot (QD)-induced fluorescence detection of bead-based DNA sandwich hybridization was studied for rapid analysis of the Bacillus spoOA gene in biofilms. Hybridization between two DNA probes and target DNA occurred and the hybridization signal was detected in a flow cytometer. To prepare the bead-DNA capture and QD-DNA detection probes, the coupling or bioconjugation reactions were carefully controlled. It was successfully demonstrated that the fluorescence response of the hybrid complex was linear in the range of 3.2-1000 nM of synthetic target DNA (R(2) = 0.97) and that the detection limit was 0.02 nM. An optimized labeling method and bead-based DNA hybridization were then applied to real PCR products from a biofilm sample with satisfactory results (R(2) = 0.94), thereby confirming that the proposed assay provides a rapid, sensitive, and specific method for Bacillus spoOA gene detection in bofilms. This approach enables multiple target detection using multicolor QD-DNA probes in a shorter time.

Photoelectronic characterization of IgG antibody molecule-quantum dot hybrid as biosensing probe.

Quantum dot (QD)-based biomolecule hybrids have recently attracted much attention in specifically identifying and labeling target proteins. In this study, QD encapsulated with immunoglobulin antibodies, as a labeling building block in biosensors, was investigated to clarify the most efficient configuration and photoluminescence behavior. Both the biological recognition capacity and photoluminescence emitting signal of the antibody-coupled nanocrystal were validated through a photoelectrical characterization procedure. Derivation of the optimum number of antibody molecules to be packed onto the QD surface yielded the highest binding capacity for the target antigen. During formation of the bioactive layer, the intrinsic photoluminescence response of the QDs significantly decreased due to photoinduced hole transfer according to their rearranged electronic structure. The thorough study of this assembly provides a validation approach for the careful titration of biosensor probes for optimal reaction kinetics. Furthermore, it contributes to the development of an effective tool for the application and interpretation of QD-based labeling techniques.

Electrochemical immunoassay using quantum dot/antibody probe for identification of cyanobacterial hepatotoxin microcystin-LR.

The presence of cyanobacterial hepatotoxins such as microcystin-LR poses health threats to humans due to their potential for causing severe physiological effects when contaminated drinking water is ingested. Here, the electrochemical detection of microcystin-LR is explored using a quantum dot/antibody (QD/Ab) probe for nanoparticle-based amplification and direct electrochemical transduction. The immunological recognition of microcystin-LR using the QD/Ab probe was amplified and converted to an electrochemical signal by measuring the cadmium ions released from QD based on square wave stripping voltammetry under optimized electrochemical factors. Whereas a qualitative analysis for microcystin-LR was achieved using the specific peak potential of the anodic voltammogram at -0.6 +/- 0.05 V, concentration of the toxin was quantified based on the charge density of the anodic peak; a dynamic range of 0.227 to 50 microg/L and limit of detection of 0.099 microg/L were obtained with high sensitivity. The extracted microcystin-LR from Microcystis aeruginosa was estimated as 1,944 microg/g of dried weight of the microorganism.

High-flux ultrafiltration membrane with open porous hydrophilic structure using dual pore formers.

This work investigated the synergistic effect of polyvinylpyrrolidone (PVP) and hydroxypropyl-beta-cyclodextrin (HP-ß-CD) as dual pore forming agents on the properties and performance of polysulfone (PSf) ultrafiltration membranes. A fixed concentration of PVP and varying concentrations of HP-ß-CD were used to prepare the membranes using the phase inversion technique. The results showed that the inclusion of these additives in the dope solution increased its thermodynamic instability and promoted instantaneous demixing. Overall, an increase was observed in the hydrophilicity, open porous structure and mechanical strength of the membranes. Cross-flow filtration tests demonstrated that the pure water permeability of the fabricated membrane was 891 LMH bar-1, about 4.37 times higher than the pristine membrane, while bovine serum albumin (BSA) rejection was relatively constant (about 93%) for all the fabricated membranes. This work proposed that the addition of HP-ß-CD and PVP as dual pore formers can produce a viable ultrafiltration membrane with improved water permeability without a middle ground on rejection potential.

Strategies for selecting indicator compounds to assess attenuation of emerging contaminants during UV advanced oxidation processes.

A ranking system for monitoring-based process control was developed to select indicator compounds that can predict the attenuation of a broader range of trace organic compounds (TOrCs) in reclaimed water by low pressure (LP) and medium pressure (MP)-UV advanced oxidation processes (AOPs). The selected TOrCs were classified into three groups depending on their relative reactivity to UV direct photolysis and •OH oxidation. Group 1 includes the photolabile TOrCs, which are easily photodegraded with no additional oxidants by either LP or MP-UV light and include acesulfame, diclofenac, and sulfamethoxazole. Group 2 consists of the moderate photodegradable compounds with high reactivity of •OH oxidation, which include benzotriazole, fluoxetine, and hydrochlorothiazide as indicator compounds for assessing LP-UV AOP and propranolol, diltiazem, and diphenhydramine for MP-UV AOP. Group 3 is photo-resistant TOrCs, but highly reactive with •OH radicals and includes carbamazepine and DEET as appropriate indicator compounds. Therefore, the proposed ranking system is expected to provide a comprehensive monitoring tool to water reuse utilities for prioritizing a list of indicators to assess the treatment efficacy of UV AOPs that allows for subsequent operational control to achieve the treatment goal. This is the first strategic framework and guidelines for building a customizable tool of process control that depend on the site-specific occurrence profile of wastewater effluents and the UV system (UV lamp spectral output and power density).

On-line sensor monitoring for chemical contaminant attenuation during UV/H2O2 advanced oxidation process.

A combination of surrogate parameters and indicator compounds were measured to predict the removal efficiency of trace organic compounds (TOrCs) using low pressure (LP)-UV/H2O2 advanced oxidation process (AOP), engaged with online sensor-based monitoring system. Thirty-nine TOrCs were evaluated in two distinct secondary wastewater effluents in terms of estimated photochemical reactivity, as a function of the rate constants of UV direct photolysis (kUV) and hydroxyl radical (OH) oxidation (kOH). The selected eighteen TOrCs were classified into three groups that served as indicator compounds: Group 1 for photo-susceptible TOrCs but with minor degradation by OH oxidation (diclofenac, fluoxetine, iohexol, iopamidol, iopromide, simazine and sulfamethoxazole); Group 2 for TOrCs susceptible to both direct photolysis and OH oxidation (benzotriazole, diphenhydramine, ibuprofen, naproxen and sucralose); and Group 3 for photo-resistant TOrCs showing dominant degradation by OH oxidation (atenolol, carbamazepine, DEET, gemfibrozil, primidone and trimethoprim). The results indicate that TOC (optical-based measurement), UVA254 or UVT254 (UV absorbance or transmittance at 254 nm), and total fluorescence can all be used as suitable on-line organic surrogate parameters to predict the attenuation of TOrCs. Furthermore, the automated real-time monitoring via on-line surrogate sensors and equipped with the developed degradation profiles between sensor response and a group of TOrCs removal can provide a diagnostic tool for process control during advanced treatment of reclaimed waters.

Multiplex competitive microbead-based flow cytometric immunoassay using quantum dot fluorescent labels.

In answer to the ever-increasing need to perform the simultaneous analysis of environmental hazards, microcarrier-based multiplex technologies show great promise. Further integration with biofunctionalized quantum dots (QDs) creates new opportunities to extend the capabilities of multicolor flow cytometry with their unique fluorescence properties. Here, we have developed a competitive microbead-based flow cytometric immunoassay using QDs fluorescent labels for simultaneous detection of two analytes, bringing the benefits of sensitive, rapid and easy-of-manipulation analytical tool for environmental contaminants. As model target compounds, the cyanobacterial toxin microcystin-LR and the polycyclic aromatic hydrocarbon compound benzo[a]pyrene were selected. The assay was carried out in two steps: the competitive immunological reaction of multiple targets using their exclusive sensing elements of QD/antibody detection probes and antigen-coated microsphere, and the subsequent flow cytometric analysis. The fluorescence of the QD-encoded microsphere was thus found to be inversely proportional to target analyte concentration. Under optimized conditions, the proposed assay performed well within 30 min for the identification and quantitative analysis of the two environmental contaminants. For microcystin-LR and benzo[a]pyrene, dose-response curves with IC(50) values of 5 µg L(-1) and 1.1 µg L(-1) and dynamic ranges of 0.52-30 µg L(-1) and 0.13-10 µg L(-1) were obtained, respectively. Recovery was 92.6-106.5% for 5 types of water samples like bottled water, tap water, surface water and seawater using only filtration as sample pretreatment.

Effects of biofouling on ion transport through cation exchange membranes and microbial fuel cell performance.

This study examines the effects of biofouling on the electrochemical properties of cation exchange membranes (CEMs), such as membrane electrical resistance (MER), specific proton conductivity (SC), and ion transport number (t(+)), in addition to on microbial fuel cell (MFC) performance. CEM biofouling using a 15.5 ± 4.6 µm biofilm was found to slightly increase the MER from 15.65 Ω cm(2) (fresh Nafion) to 19.1 Ω cm(2), whereas an increase of almost two times was achieved when the electrolyte was changed from deionized water to an anolyte containing a high cation concentration supporting bacterial growth. The simple physical cleaning of CEMs had little effect on the Coulombic efficiency (CE), whereas replacing a biofouled CEM with new one resulted in considerable increase of up to 59.3%, compared to 45.1% for a biofouled membrane. These results clearly suggest the internal resistance increase of MFC was mainly caused by the sulfonate functional groups of CEM being occupied with cations contained in the anolyte, rather than biofouling itself.

Formation of hazardous inorganic by-products during electrolysis of seawater as a disinfection process for desalination.

From our previous study, an electrochemical process was determined to be a promising tool for disinfection in a seawater desalination system, but an investigation on the production of several hazardous by-products is still required. In this study, a more intensive exploration of the formation patterns of perchlorate and bromate during the electrolysis of seawater was conducted. In addition, the rejection efficiencies of the targeted by-products by membrane processes (microfiltration and seawater reverse osmosis) were investigated to uncover the concentrations remaining in the final product from a membrane-based seawater desalination system for the production of drinking water. On the electrolysis of seawater, perchlorate did not provoke any problem due to the low concentrations formed, but bromate was produced at a much higher level, resulting in critical limitation in the application of the electrochemical process to the desalination of seawater. Even though the formed bromate was rejected via microfiltration and reverse osmosis during the 1st and 2nd passes, the residual concentration was a few orders of magnitude higher than the USEPA regulation. Consequently, it was concluded that the application of the electrochemical process to seawater desalination cannot be recommended without the control of bromate.