DNA extraction yield from drinking water distribution systems and premise plumbing is a key metric for any downstream analysis such as 16S amplicon or metagenomics sequencing. This research aimed to optimize DNA yield from low-biomass (chlorinated) reverse osmosis-produced tap water by evaluating the impact of different factors during the DNA extraction procedure. The factors examined are (1) the impact of membrane materials and their pore sizes; (2) the impact of different cell densities; and (3) an alternative method for enhancing DNA yield via incubation (no nutrient spiking). DNA from a one-liter sampling volume of RO tap water with varying bacterial cell densities was extracted with five different filter membranes (mixed ester cellulose 0.2 μm, polycarbonate 0.2 μm, polyethersulfone 0.2 and 0.1 μm, polyvinylidene fluoride 0.1 μm) for biomass filtration. Our results show that (i) smaller membrane pore size solely did not increase the DNA yield of low-biomass RO tap water; (ii) the DNA yield was proportional to the cell density and substantially dependent on the filter membrane properties (i.e., the membrane materials and their pore sizes); (iii) by using our optimized DNA extraction protocol, we found that polycarbonate filter membrane with 0.2 μm pore size markedly outperformed in terms of quantity (DNA yield) and quality (background level of 16S gene copy number) of recovered microbial DNA; and finally, (iv) for one-liter sampling volume, incubation strategy enhanced the DNA yield and enabled accurate identification of the core members (i.e., Porphyrobacter and Blastomonas as the most abundant indicator taxa) of the bacterial community in low-biomass RO tap water. Importantly, incorporating multiple controls is crucial to distinguish between contaminant/artefactual and true taxa in amplicon sequencing studies of low-biomass RO tap water.

Membrane modules for seawater desalination are becoming increasingly important for obtaining clean water with the rising global water scarcity. The productivity of membrane modules is compromised by biofouling on the membrane and feed spacer. Biofouling development can be mitigated by modification of the spacer or membrane surface. The purpose of the present study is to evaluate the impact of surface-modified feed spacers on the cleaning performance of spiral-wound membrane filtration systems. After cold-plasma treatment, the feed spacers were modified with various combinations of polydopamine (PDA) and silver nanoparticles (AgNP). To compare the cleaning performance of the modified and unmodified spacers, membrane fouling simulators containing nanofiltration membranes and feed spacers were tested under industrially representative conditions: two full cycles involving biofilm development followed by cleaning-in-place (CIP). The modified spacers significantly improved the CIP efficiency when compared with that of the unmodified feed spacer. The highest CIP efficiency was obtained for the PDA–AgNP-coated spacers, which removed >90 % of the biomass. The PDA layers remained undetached during the CIP process, and the amounts of AgNP decreased without affecting the CIP effectiveness during consecutive operational cycles. The results demonstrate that CIP should be included in biofouling tests to evaluate the full potential of surface modifications and suggest that hydrophilic and biocidal spacer surface coatings can significantly improve the CIP effectiveness, thereby considerably reducing the CIP frequency and operational costs.

This study describes a novel integration of aerobic granular sludge (AGS) with a gravity-driven membrane (GDM) system at a pilot scale with a treatment capacity of approximately 150 L per day to treat raw domestic wastewater. The treatment performance and energy consumption of the AGS-GDM system were compared to the neighboring full-scale aerobic membrane bioreactor (AeMBR), treating the same wastewater at about 4000(±500) m³ per day. The AGS-GDM system demonstrated superior nutrient (nitrogen and phosphorus) removal as compared to the AeMBR. The GDM unit was continuously supplied with AGS-treated effluent. The GDM unit started with high [ >20 L per m² per h (LMH) ] flux, which gradually declined. The flux remained quite stable after 15 days reaching 3 LMH after 35 days without any physical or chemical cleaning. Our results suggest that AGS-GDM is a viable technology for decentralized wastewater treatment and reuse in water-scarce regions. The AGS-GDM could easily replace conventional AeMBR technology in the wastewater treatment and reclamation market.

Early fouling warning is important for the economical operation of membrane separation systems. In parallel multi-channel flow systems, flow re-distribution between channels due to fouling is often associated with maloperation. In the current research we use low magnetic field NMR to monitor multi-fiber hollow fiber membrane modules undergoing a fouling-cleaning cycle and show that rapid detection of fouling is possible by detecting the loss of signal coherence associated with flow re-distribution within the 401 hollow fiber membrane module. This effect is demonstrated to be both reproducible, and reversible via membrane cleaning. The results demonstrate a strong correlation between the coherence signal magnitude and the number of fibers fouled. This may be used in practice for high sensitivity early warning, and to monitor the efficiency of cleaning. This approach may also be particularly useful in the case of detecting residual fouling after cleaning, evidenced in this research by significant flow re-distribution between the before fouling and after cleaning signal coherence.

In this study non-invasive low field magnetic resonance imaging (MRI) technology was used to monitor fouling induced changes in fiber-by-fiber hydrodynamics inside a multi-fiber hollow fiber membrane module containing 401 fibers. Using structural and velocity images the fouling evolution of these membrane modules were shown to exhibit distinct trends in fiber-by-fiber volumetric flow, with increasing fouling causing a decrease in the number of flow active fibers. This study shows that the fouling rate is not evenly distributed over the parallel fibers, which results in a broadening of the fiber to fiber flowrate distribution. During cleaning, this distribution is initially broadened further, as relatively clean fibers are cleaned more rapidly compared to clogged fibers. By tracking the volumetric flow rate of individual fibers inside the modules during the fouling-cleaning cycle it was possible to observe a fouling memory-like effect with residual fouling occurring preferentially at the outer edge of the fiber bundle during repeated fouling-cleaning cycle. These results demonstrate the ability of MRI velocity imaging to quantitatively monitor these effects which are important when testing the effectiveness of cleaning protocols due to the long term effect that residual fouling and memory-like effect may have on the operation of membrane modules.

In this study, a biofouling index based on the relative pressure drop is presented to quantitatively evaluate the amount of fouling in spacer-filled membrane filtration channels. The biofouling index was defined as the inverse of the time to reach a relative pressure drop of 100% and can be interpreted as a fouling rate or cleaning frequency. The index was applied to evaluate biofilm growth in membrane fouling simulators with reverse osmosis membranes and commercial feed spacers operated with different feed water nutrient concentrations and crossflow velocities. Biofilm accumulation on the membrane and feed spacer was characterized in situ using optical coherence tomography. We showed that the biofouling index is directly related to the volume of biofouling independent of the applied crossflow velocity and a suitable tool for improved quantitative comparison of the biofouling rate. Furthermore our results suggest that the pressure drop is better described as function of the velocity at the perimeter of a spacer cell instead of the average velocity in the channel. Although the biofouling index is developed for biofouling, the index may be applied to quantitatively assess mitigation strategies in spacer filled channels for a wider range of fouling types.

Monitoring the changes that occur to water during distribution is vital to ensure water safety. In this study, the biological stability of reverse osmosis (RO) produced drinking water, characterized by low cell concentration and low assimilable organic carbon, in combination with chlorine disinfection was investigated. Water quality at several locations throughout the existing distribution network was monitored to investigate whether microbial water quality changes can be identified. Results revealed that the water leaving the plant had an average bacterial cell concentration of 10³ cells/mL. A 0.5–1.5 log increase in bacterial cell concentration was observed at locations in the network. The residual disinfectant was largely dissipated in the network from 0.5 mg/L at the treatment plant to less than 0.1 mg/L in the network locations. The simulative study involving miniature distribution networks, mimicking the dynamics of a distribution network, fed with the RO produced chlorinated and non-chlorinated drinking water revealed that distributing RO produced water without residual disinfection, especially at high water temperatures (25–30 °C), poses a higher chance for water quality change. Within six months of operation of the miniature network fed with unchlorinated RO produced water, the adenosine triphosphate (ATP) and total cell concentration (TCC) in the pipe biofilm were 4 × 10² pg ATP/cm² and 1 × 10⁷ cells/ cm². The low bacterial cell concentration and organic carbon concentration in the RO-produced water did not prevent biofilm development inside the network with and without residual chlorine. The bacterial community analysis using 16S ribosomal RNA (rRNA) gene sequencing revealed that mesophilic bacteria with higher temperature tolerance and bacteria associated with oligotrophic, nutrient-poor conditions dominated the biofilm, with no indication of the existence of opportunistic pathogenic species. However, chlorination selected against most bacterial groups and the bacterial community that remained was mainly the bacteria capable of surviving disinfection regimes. Biofilms that developed in the presence of chlorine contained species classified as opportunistic pathogens. These biofilms have an impact on shaping the water quality received at the consumer tap. The presence of these bacteria on its own is not a health risk indicator; viability assessment and qPCRs targeting genes specific to the opportunistic pathogens as well as quantitative microbiological risk assessment (QMRA) should be included to assess the risk. The results from this study highlight the importance of implementing multiple barriers to ensure water safety. Changes in water quality detected even when high-quality disinfected RO-produced water is distributed highlight microbiological challenges that chlorinated systems endure, especially at high water temperatures.

Nutrient limitation has been proposed as a biofouling control strategy for membrane systems. However, the impact of permeation on biofilm development under phosphorus-limited and enriched conditions is poorly understood. This study analyzed biofilm development in membrane fouling simulators (MFSs) with and without permeation supplied with water varying dosed phosphorus concentrations (0 and 25 µg P·L⁻¹). The MFSs operated under permeation conditions were run at a constant flux of 15.6 L·m^2·h−1 for 4.7 days. Feed channel pressure drop, transmembrane pressure, and flux were used as performance indicators. Optical coherence tomography (OCT) images and biomass quantification were used to analyze the developed biofilms. The total phosphorus concentration that accumulated on the membrane and spacer was quantified by using microwave digestion and inductively coupled plasma atomic emission spectroscopy (ICP-OES). Results show that permeation impacts biofilm development depending on nutrient condition with a stronger impact at low P concentration (pressure drop increase: 282%; flux decline: 11%) compared to a higher P condition (pressure drop increase: 206%; flux decline: 2%). The biofilm that developed at 0 µg P·L⁻¹ under permeation conditions resulted in a higher performance decline due to biofilm localization and spread in the MFS. A thicker biofilm developed on the membrane for biofilms grown at 0 µg P·L⁻¹ under permeation conditions, causing a stronger effect on flux decline (11%) compared to non-permeation conditions (5%). The difference in the biofilm thickness on the membrane was attributed to a higher phosphorus concentration in the membrane biofilm under permeation conditions. Permeation has an impact on biofilm development and, therefore, should not be excluded in biofouling studies.

The application of membrane technology for water treatment and reuse is hampered by the development of a microbial biofilm. Biofilm growth in micro-and ultrafiltration (MF/UF) membrane modules, on both the membrane surface and feed spacer, can form a secondary membrane and exert resistance to permeation and crossflow, increasing energy demand and decreasing permeate quantity and quality. In recent years, exhaustive efforts were made to understand the chemical, structural and hydraulic characteristics of membrane biofilms. In this review, we critically assess which specific structural features of membrane biofilms exert resistance to forced water passage in MF/UF membranes systems applied to water and wastewater treatment, and how biofilm physical structure can be engineered by process operation to impose less hydraulic resistance (“below-the-pain threshold”). Counter-intuitively, biofilms with greater thickness do not always cause a higher hydraulic resistance than thinner biofilms. Dense biofilms, however, had consistently higher hydraulic resistances compared to less dense biofilms. The mechanism by which density exerts hydraulic resistance is reported in the literature to be dependant on the biofilms’ internal packing structure and EPS chemical composition (e.g., porosity, polymer concentration). Current reports of internal porosity in membrane biofilms are not supported by adequate experimental evidence or by a reliable methodology, limiting a unified understanding of biofilm internal structure. Identifying the dependency of hydraulic resistance on biofilm density invites efforts to control the hydraulic resistance of membrane biofilms by engineering internal biofilm structure. Regulation of biofilm internal structure is possible by alteration of key determinants such as feed water nutrient composition/concentration, hydraulic shear stress and resistance and can engineer biofilm structural development to decrease density and therein hydraulic resistance. Future efforts should seek to determine the extent to which the concept of “biofilm engineering” can be extended to other biofilm parameters such as mechanical stability and the implication for biofilm control/removal in engineered water systems (e.g., pipelines and/or, cooling towers) susceptible to biofouling.

Natural Deep Eutectic Solvents (NADES) are composed of supramolecular interactions of two or more natural compounds, such as organic acids, sugars, and amino acids, and they are being used as a new media alternative to conventional solvents. In this study, a new application of NADES is presented as a possible technology for biofilm structural breaker in complex systems since the current solvents used for biofilm cleaning and extraction of biofilm components use hazardous solutions. The NADES (betaine:urea:water and lactic acid:glucose:water) were analyzed before and after the biofilm treatment by attenuated total reflection Fourier-transform infrared spectroscopy and fluorescence excitation-emission matrix spectroscopy. Our results indicate that the green solvents could solubilize up to ≈70 percent of the main components of the biofilms extracellular matrix. The solubilization of the biomolecules weakened the biofilm structure, which could enhance the biofilm solubilization and removal. The NADES have the potential to be an environment-friendly, green solvent to extract valuable compounds and break the main structure from the biofilm, leading to a greener method for extracellular polymeric substance (EPS) extraction and biofilm treatment in various water treatment systems.

Temperature change over the length of heat exchangers might be an important factor affect-ing biofouling. This research aimed at assessing the impact of temperature on biofilm accumulation and composition with respect to bacterial community and extracellular polymeric substances. Two identical laboratory-scale plate heat exchanger modules were developed and tested. Tap water sup-plemented with nutrients was fed to the two modules to enhance biofilm formation. One “reference” module was kept at 20.0 ± 1.4^◦ C and one “heated” module was operated with a counter-flow hot water stream resulting in a bulk water gradient from 20 to 27^◦ C. Biofilms were grown during 40 days, sampled, and characterized using 16S rRNA gene amplicon sequencing, EPS extraction, FTIR, protein and polysaccharide quantifications. The experiments were performed in consecutive triplicate. Monitoring of heat transfer resistance in the heated module displayed a replicable biofilm growth profile. The module was shown suitable to study the impact of temperature on biofouling formation. Biofilm analyses revealed: (i) comparable amounts of biofilms and EPS yield in the reference and heated modules, (ii) a significantly different protein to polysaccharide ratio in the EPS of the reference (5.4 ± 1.0%) and heated modules (7.8 ± 2.1%), caused by a relatively lower extracellular sugar production at elevated temperatures, and (iii) a strong shift in bacterial community composition with increasing temperature. The outcomes of the study, therefore, suggest that heat induces a change in biofilm bacterial community members and EPS composition, which should be taken into consideration when investigating heat exchanger biofouling and cleaning strategies. Research potential and optimization of the heat exchanger modules are discussed.

Biofouling is a problem that hinders sustainable membrane-based desalination and the stratification of bacterial populations over the biofilm’s height is suggested to compromise the efficiency of cleaning strategies. Some studies reported a base biofilm layer attached to the membrane that is harder to remove. Previous research suggested limiting the concentration of phosphorus in the feed water as a biofouling control strategy. However, the existence of bacterial communities growing under phosphorus-limiting conditions and communities remaining after cleaning is unknown. This study analyzes the bacterial communities developed in biofilms grown in membrane fouling simula-tors (MFSs) supplied with water with three dosed phosphorus conditions at a constant biodegradable carbon concentration. After biofilm development, biofilm was removed using forward flushing (an easy-to-implement and environmentally friendly method) by increasing the crossflow velocity for one hour. We demonstrate that small changes in phosphorus concentration in the feed water led to (i) different microbial compositions and (ii) different bacterial-cells-to-EPS ratios, while (iii) similar bacterial biofilm populations remained after forward flushing, suggesting a homogenous bacterial community composition along the biofilm height. This study represents an exciting advance towards greener desalination by applying non-expensive physical cleaning methods while manipulating feed water nutrient conditions to prolong membrane system performance and enhance membrane cleanability.

Biofouling is one of the major factors causing decline in membrane performance in reverse osmosis (RO) plants, and perhaps the biggest hurdle of membrane technology. Chemical cleaning is periodically carried out at RO membrane installations aiming to restore membrane performance. Typical cleaning agents used in the water treatment industry include sodium hydroxide (NaOH) and hydrochloric acid (HCl) in sequence. Rapid biofilm regrowth and related membrane performance decline after conventional chemical cleaning is a routinely observed phenomenon due to the inefficient removal of biomass from membrane modules. Since extracellular polymeric substances (EPS) make up the strongest and predominant structural framework of biofilms, disintegration of the EPS matrix should be the main target for enhanced biomass removal. Previously, we demonstrated at lab-scale the use of concentrated urea as a chemical cleaning agent for RO membrane systems. The protein denaturation property of urea was exploited to solubilize the proteinaceous foulants, weakening the EPS layer, resulting in enhanced biomass solubilization and removal from RO membrane systems. In this work, we investigated the impact of repeated chemical cleaning cycles with urea/HCl as well as NaOH/HCl on biomass removal and the potential adaptation of the biofilm microbial community. Chemical cleaning with urea/HCl was consistently more effective than NaOH/HCl cleaning over 6 cleaning and regrowth cycles. At the end of the 6 cleaning cycles, the percent reduction was 35% and 41% in feed channel pressure drop, 50% and 70% in total organic carbon, 30% and 40% in EPS proteins, and 40% and 66% in the peak intensities of protein-like matter, after NaOH/HCl cleaning and Urea/HCl cleaning, respectively. 16S ribosomal RNA (rRNA) gene sequencing of the biofilm microbial community revealed that urea cleaning does not select for key biofouling families such as Sphingomonadaceae and Xanthomonadaceae that are known to survive conventional chemical cleaning and produce adhesive EPS. This study reaffirmed that urea possesses all the desirable properties of a chemical cleaning agent, i.e., it dissolves the existing fouling layer, delays fresh fouling accumulation by inhibiting the production of a more viscous EPS, does not cause damage to the membranes, is chemically stable, and environmentally friendly as it can be recycled for cleaning.

The manner in which membrane-fouling experiments are conducted and how fouling performance data are represented have a strong impact on both how the data are interpreted and on the conclusions that may be drawn. We provide a couple of examples to prove that it is possible to obtain misleading conclusions from commonly used representations of fouling data. Although the illustrative example revolves around dead-end ultrafiltration, the underlying principles are applicable to a wider range of membrane processes. When choosing the experimental conditions and how to represent fouling data, there are three main factors that should be considered: (I) the foulant mass is principally related to the filtered volume; (II) the filtration flux can exacerbate fouling effects (e.g., concentration polarization and cake compression); and (III) the practice of normalization, as in dividing by an initial value, disregards the difference in driving force and divides the fouling effect by different numbers. Thus, a bias may occur that favors the experimental condition with the lower filtration flux and the less-permeable membrane. It is recommended to: (I) avoid relative fouling performance indicators, such as relative flux decline (J/J0); (II) use resistance vs. specific volume; and (III) use flux-controlled experiments for fouling performance evaluation.

Desalination technology based on Reverse Osmosis (RO) membrane filtration has been resorted to provide high-quality drinking water. RO produced drinking water is characterized by a low bacterial cell concentration. Monitoring microbial quality and ensuring membrane-treated water safety has taken advantage of the rapid development of DNA-based techniques. However, the DNA extraction process from RO-based drinking water samples needs to be evaluated regarding the biomass amount (filtration volume) and residual disinfectant such as chlorine, as it can affect the DNA yield. We assessed the DNA recovery applied in drinking water microbiome studies as a function of (i) different filtration volumes, (ii) presence and absence of residual chlorine, and (iii) the addition of a known Escherichia coli concentration into the (sterile and non-sterile, chlorinated and dechlorinated) tap water prior filtration, and directly onto the (0.2 μm pore size, 47 mm diameter) mixed ester cellulose membrane filters without and after tap water filtration. Our findings demonstrated that the co-occurrence of residual chlorine and low biomass/cell density water samples (RO-treated water with a total cell concentration ranging between 2.47 × 10²–1.5 × 10³ cells/mL) failed to provide sufficient DNA quantity (below the threshold concentration required for sequencing-based procedures) irrespective of filtration volumes used (4, 20, 40, 60 L) and even after performing dechlorination. After exposure to tap water containing residual chlorine (0.2 mg/L), we observed a significant reduction of E. coli cell concentration and the degradation of its DNA (DNA yield was below detection limit) at a lower disinfectant level compared to what was previously reported, indicating that free-living bacteria and their DNA present in the drinking water are subject to the same conditions. The membrane spiking experiment confirmed no significant impact from any potential inhibitors (e.g. organic/inorganic components) present in the drinking water matrix on DNA extraction yield. We found that very low DNA content is likely to be the norm in chlorinated drinking water that gives hindsight to its limitation in providing robust results for any downstream molecular analyses for microbiome surveys. We advise that measurement of DNA yield is a necessary first step in chlorinated drinking water distribution systems (DWDSs) before conducting any downstream omics analyses such as amplicon sequencing to avoid inaccurate interpretations of results based on very low DNA content. This study expands a substantial source of bias in using DNA-based methods for low biomass samples typical in chlorinated DWDSs. Suggestions are provided for DNA-based research in drinking water with residual disinfectant.

A critical problem in seawater reverse osmosis (RO) filtration processes is biofilm accumulation, which reduces system performance and increases energy requirements. As a result, membrane systems need to be periodically cleaned by combining chemical and physical protocols. Nutrient limitation in the feed water is a strategy to control biofilm formation, lengthening stable membrane system performance. However, the cleanability of biofilms developed under various feed water nutrient conditions is not well understood. This study analyzes the removal efficiency of biofilms grown in membrane fouling simulators (MFSs) supplied with water varying in phosphorus concentrations (3 and 6 μg P·L^-1 and with constant biodegradable carbon concentration) by applying hydraulic cleaning after a defined 140% increase in the feed channel pressure drop, through increasing the cross-flow velocity from 0.18 m s^-1 to 0.35 m s^-1 for 1 h. The two phosphorus concentrations (3 and 6 μg P·L^-1) simulate the RO feed water without and with the addition of a phosphorus-based antiscalant, respectively, and were chosen based on measurements at a full-scale seawater RO desalination plant. Biomass quantification parameters performed after membrane autopsies such as total cell count, adenosine triphosphate, total organic carbon, and extracellular polymeric substances were used along with feed channel pressure drop measurements to evaluate biofilm removal efficiency. The outlet water during hydraulic cleaning (1 h) was collected and characterized as well. Optical coherence tomography images were taken before and after hydraulic cleaning for visualization of biofilm morphology. Biofilms grown at 3 μg P·L^-1 had an enhanced hydraulic cleanability compared to biofilms grown at 6 μg P·L^-1. The higher detachment for biofilms grown at a lower phosphorus concentration was explained by more soluble polymers in the EPS, resulting in a lower biofilm cohesive and adhesive strength. This study confirms that manipulating the feed water nutrient composition can engineer a biofilm that is easier to remove, shifting research focus towards biofilm engineering and more sustainable cleaning strategies.

A novel magnetic resonance measurement (MRM) protocol for non-invasive monitoring of fouling in spiral wound reverse osmosis (SWRO) membrane modules is demonstrated. Sodium alginate was used to progressively foul a commercial SWRO membrane at industrially relevant operating conditions in a circulating flow loop. The MRM protocol showcased the following: (i) earlier, more sensitive detection and quantification of fouling in the membrane module compared to feed-channel pressure drop. This was achieved using appropriate detection of the total nuclear magnetic resonance (NMR) signal. (ii) 2D cross-sectional imaging of the location of the accumulated foulant material; this was preferentially located adjacent to the membrane spacer sheet nodes, which was subsequently confirmed by a module autopsy. This image contrast, which could also readily differentiate the membrane, feed spacer and permeate spacer regions, was realised based on differences in the NMR relaxation parameter, T_2,eff. (iii) High frequency acquisition of 2D cross-sectional velocity images of the module revealing very localised flow channelling in response to gradual foulant accumulation which impacted significantly on the flow pattern within the central permeate tube. Collectively this NMR/MRI measurement protocol provides a powerful analysis tool for the evolution of fouling in such complex modules, thus ultimately enabling more informed module design.

This study systematically investigated the effect of organic micropollutants (OMPs) on biofouling in forward osmosis (FO) integrating wastewater treatment and seawater dilution. Synthetic seawater (0.6 M sodium chloride) was used as a draw solution and synthetic municipal wastewater as a feed solution. To evaluate the impact of OMPs in a replicate parallel study, wastewater was supplemented with a mixture of 7 OMPs (OMPs-feed) and without OMPs (control) during 8 batch filtration cycles with feed and draw solution replacement after each filtration. The FO performance (water flux), development and microbial composition properties of biofilm layers on the wastewater side of the FO membrane were studied. Compared to the control without OMPs, the FO fed with OMPs containing wastewater showed (i) initially the same water flux and flux decline during the first filtration cycle, (ii) with increasing filtration cycle a lower flux decline and (iii) lower concentrations for the total cells, ATP, EPS carbohydrates and proteins in biofilm layers, and (iv) a lower diversity of the biofilm microbial community composition (indicating selective pressure) and (v) increasing rejection of 6 of the 7 OMPs. In essence, biofouling on the FO membrane showed (i) a lower flux decline in the presence of OMPs in the feed water and (ii) a higher OMPs rejection, both illustrating better membrane performance. This study has a significant implication for optimizing osmotic dilution in terms of FO operation and OMPs rejection.

Efficient control of pathogenic bacteria, specifically Legionella pneumophila, is one of the main concerns when operating industrial cooling towers. Common practices to limit proliferation involves use of disinfectants, leading to formation of disinfection by-product and increase in water corrosiveness. A disinfectant-free Legionella control method would make the industry more environmentally friendly. A pilot-scale cooling tower (1 m3/h) operated with demineralized water was used to investigate the potential of high-pH conditioning as a disinfectant-free alternative for control of L. pneumophila and other pathogens. One control experiment was performed under standard full-scale operation involving sodium hypochlorite dosage. Thereafter 3 alkaline pHs of the cooling water were tested: 9.0, 9.4 and 9.6. The tests lasted between 25 and 35 days. The cooling water from the basins were analysed for total cell count by flow cytometry, L. pneumophila concentration by plate count and occasional qPCR analyses targeting the mip-gene, bacterial and eukaryotic community analyses with 16S and 18S rRNA gene amplicon sequencing, relative abundance of eukaryotic to prokaryotic DNA by qPCR of the 16S and 18S rRNA gene. The L. pneumophila analyses showed considerable growth at pH 9.0 and pH 9.4 but was maintained below detection limit (< 100 CFU/L) at pH 9.6 without disinfection. Interestingly, the results correlated with the overall abundance of protozoa in the water samples but not directly with the relative abundance of specific reported protozoan hosts of Legionella. The pathogenicity based on 16S rRNA gene amplicon sequencing of the cooling water DNA decreased with increasing pH with a strong decline between pH 9.0 and pH 9.4, from 7.1% to 1.6% of relative abundance of pathogenic genera respectively. A strong shift in microbiome was observed between each tested pH and reproducibility of the experiment at pH 9.6 was confirmed with a duplicate test lasting 80 days. High-pH conditioning ≥ 9.6 is therefore considered as an efficient disinfectant-free cooling tower operation for control of pathogenicity, including L. pneumophila.