The textile industry has long been associated with significant environmental challenges due to the generation of recalcitrant wastewater, containing complex chemical mixture that pose severe threats to ecosystems and human health. This study focuses on the role of pH adjustment in improving the pretreatment process for direct contact membrane distillation (DCMD) applied to real textile wastewater. By implementing a pH adjustment step (pH 6.14, 7.40, and 9.06) prior to sedimentation and filtration, the pretreatment process was significantly enhanced, reducing wetting, and improving permeate quality. GC–MS analysis identified specific organic molecules causing wetting, including volatile organic acids and alcohol derivatives, revealing that the rejection mechanism is primarily driven by the relationship between the wastewater pH and the pKa of these compounds. Adjusting the pH above the pKa converts these acidic contaminants into ionic, non-volatile forms, effectively preventing their passage into the permeate. This study highlights the importance of pH optimization in advancing DCMD as a sustainable solution for textile wastewater treatment. The proposed approach aligns with circular economy principles, enabling water reuse in textile processes, reducing freshwater consumption, and minimizing environmental discharge.

Designing robust drinking water treatment schemes for eutrophic sources requires shifting from considering each treatment step separately to considering the full treatment process as a connected system. This study evaluated how treatment configuration and arrangement influence microbial community dynamics, organic carbon removal, and biological stability in a full-scale drinking water treatment plant. A Dutch treatment plant was monitored, operating two parallel lines: one conventional (coagulation, sedimentation, and rapid sand filtration) and one advanced (ion exchange, ceramic microfiltration, and advanced oxidation), both converging into granular activated carbon (GAC) filtration. Microbial and chemical water quality was assessed across treatment stages and seasons. This plant experiences periods of discoloration, taste, and odor issues, and an exceedance of Aeromonas counts in the distribution network. Advanced oxidation achieved a high bacterial cell inactivation (~90%); however, it significantly increased assimilable organic carbon (AOC) (300–900% increase), challenging biological stability. GAC filtration partially reduced AOC levels (from 70 μg Ac-C/L to 12 μg Ac-C/L) but also supported dense (105 cells/mL) and diverse microbial communities (Shannon diversity index 5.83). Moreover, Gammaproteobacteria, which harbor opportunistic pathogens such as Aeromonas, persisted during the treatment. Archaea were highly sensitive to oxidative and physical stress, leading to reduced diversity downstream. Beta diversity analysis revealed that treatment configuration, rather than seasonality, governed the community composition. The findings highlight that treatment arrangement, oxidation, GAC operation, and organic and microbial loads critically influence biological stability. This study proposes integrated strategies to achieve resilient and biologically stable drinking water production when utilizing complex water sources such as eutrophic lakes.

Reverse osmosis (RO) desalination is the leading technology for industrial and municipal water production in water-stressed regions. While developing chemical-free scaling control strategies helps mitigate the environmental impact of brine discharge, it also increases the risk of membrane scaling due to high salt concentrations. Establishing methods for early detection and localization of scaling is essential, as well as understanding the impact on key operational parameters. This study evaluated optical coherence tomography (OCT) for real-time monitoring of growth- and deposition-driven gypsum fouling in RO systems. Membrane fouling simulators were operated under constant flux conditions using unsaturated and supersaturated synthetic water solutions. Real-time monitoring of operational parameters revealed that growth and deposition fouling had a greater impact on transmembrane pressure than pressure drop increase. OCT imaging visualized scaling progression, with optical and SEM imaging confirming distinct morphologies: sharp, translucent crystals in growth-driven scaling and a white, amorphous fouling layer in deposition. Data processing further provided quantitative assessment of area coverage and fouling volume, with membrane autopsy indicating higher porosity in the deposition case. Crystal detection from OCT imaging evidenced sensitivity for early-stage scaling detection. In the growth case, a strong correlation was observed between initial crystal formation and regions of maximum saturation index, as revealed by CFD with multicomponent solute transport simulations. The variation in induction time across detection methods highlights the importance of sensitivity of monitoring techniques, positioning OCT as a valuable tool for early scaling detection, before conventional indicators point out to significant scaling.

This study addresses the pervasive challenge of biofouling in seawater desalination systems, which compromises membrane performance and longevity, by introducing a multifunctional PDA-SP-cTA coating. This bio-inspired coating effectively mitigates biofouling in seawater reverse osmosis systems without requiring biocide. The coating is applied to both hydrophilic polyamide membranes and hydrophobic polypropylene feed spacers through in-situ and ex-situ polymer deposition methods, involving a single-step process with polydopamine and sodium-periodate, followed by surface tailoring with citric acid-blended tannic acid. Extensive surface characterization, primarily conducted on polypropylene feed spacers, confirms coating deposition. Antibiofouling properties are evaluated through long-term biofouling tests simulating industrial conditions. The findings demonstrate that the ex-situ applied coating significantly reduces relative feed channel pressure drop increase due to biofilm growth by 75 % and lowers biomass accumulation (88 % total cell counts, 70 % adenosine-triphosphate, 91 % carbohydrates, and 69 % proteins). The coating inhibits the colonization of biofouling-causing bacterial genus Alteromonas, drastically decreases active bacterial gene copy numbers, and alters microbial composition, leading to reduced biofilm viability and loosely attached biofilms that could enhance cleaning efficiency. This comprehensive study encompasses the entire process from the strategic selection and systematic characterization of the coating to extensive biofouling tests and stability assessments offering a holistic solution to combat biofouling without biocides. With demonstrated durability and stability across various pH conditions over time, this coating could be a widely applicable and scalable solution for biofouling mitigation in diverse industrial contexts.

Seawater reverse osmosis (SWRO) membrane systems face inevitable performance decline due to biofouling, which imposes significant economic costs, accounting up to 25 % of water production costs in desalination plants. Current industry practices primarily rely on chemical cleaning treatments to restore membrane performance. However, these methods involve substantial expenses related to chemical acquisition, storage, and transportation, extended plant downtimes, and premature membrane replacement due to reduced lifespan. Additionally, the disposal of chemical waste raises serious environmental concerns. Micro-nano bubbles (MNBs), consisting of gas-filled cavities (from <1 μm to 5 μm in diameter), have emerged as a promising alternative for biofouling control. This study evaluates the performance of the air-filled (AMNBs) and CO2-nucleated bubbles (N_CO2) as curative cleaning-in-place (CIP) treatments and preventive daily treatments under conditions representative of SWRO systems. Using membrane fouling simulators (MFSs) and pressure drop as a performance indicator, curative AMNBs, and N_CO2 treatments achieved 49 %–56 % pressure drop recovery, comparable to conventional chemical cleaning (51 %) and significantly outperforming hydraulic flushing (24 %). Optical coherence tomography (OCT) imaging and biomass analyses confirmed these findings, revealing effective biofilm removal due to MNB's action. Preventive treatments demonstrated that Nucleated CO2 bubbles delayed performance decline by 123 %, AMNBs by 95 %, and hydraulic flushing by only 15 % compared to controls. OCT imaging and membrane biomass analysis confirmed reduced biofilm growth and biomass accumulation in bubble-treated systems. These results indicate that MNB technologies hold great potential as a sustainable and eco-friendly alternative to chemical cleaning for biofouling management in SWRO systems.

This case study investigated the effectiveness of Direct Contact Membrane Distillation (DCMD) in treating real textile wastewater. Textile wastewater treatment presents a critical challenge in the field of environmental sustainability, requiring innovative approaches for its treatment to mitigate adverse impacts on ecosystems. DCMD emerges as a promising solution for the treatment and reuse of textile wastewater. However, the intricate composition of real textile wastewater represents a major bottleneck for the process, as the effectiveness of DCMD is influenced by numerous factors, complicating its application. In this study, experiments with an untreated sample demonstrate the detrimental impact of suspended solids on membrane performance. The application of simple pretreatment steps prior to DCMD, involving sedimentation and filtration, substantially enhanced the quality of the permeate, resulting in 100 % color removal, 99.99 % turbidity removal, and considerable removal rates for Chemical Oxygen Demand (COD) and Total Organic Carbon (TOC). Nevertheless, wetting remained a significant issue, as evidenced by the persistence of commonly used volatile organic contaminants and surfactants in the textile industry detected within the permeate. The findings in this case study reinforce that DCMD holds promise for textile wastewater treatment but emphasize the necessity of pretreatment and wetting mitigation strategies to fully unlock its potential. This research offers crucial insights for future MD applications in addressing the complexities of textile wastewater treatment.

Domestic showers are critical points of human exposure to microbial biofilms, which may harbor opportunistic pathogens such as Legionella spp. and nontuberculous Mycobacterium. However, biofilm development in reverse osmosis (RO)-treated drinking water systems remains poorly understood. We tested whether shower plumbing material (flexible polymer hose versus showerhead with inline polyethersulfone filter) and seasonal water variations influence biofilm community assembly. In a controlled field study, commercial shower systems were deployed in households supplied with RO-treated tap water from the KAUST Seawater Desalination Plant; biofilm samples were collected from hoses and filters over 3–17 months. Flow cytometry and 16S rRNA gene amplicon sequencing characterized microbial abundance, diversity, and taxonomic composition. We found that alpha diversity, measured by observed OTUs, was uniformly low, reflecting ultra-low biomass in RO-treated tap water. Beta diversity analyses revealed clear clustering by material type, with hoses exhibiting greater richness and evenness than filters. Core taxa—Pelomonas, Blastomonas, and Porphyrobacter—dominated both biofilm types, suggesting adaptation to low-nutrient, chlorinated conditions. Overall, our results demonstrate that ultra-low-nutrient RO tap water still supports the formation of material-driven, low-diversity biofilms dominated by oligotrophic taxa, underscoring plumbing-material choice as a critical factor for safeguarding shower water quality. These findings advance our understanding of biofilm ecology in RO-treated systems, informing strategies to mitigate potential health risks in shower 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.

Biofouling poses a significant challenge to reverse osmosis (RO) membrane systems, necessitating timely detection for effective control. This study evaluated the efficacy of flow cytometry (FCM) for early biofilm detection in comparison to conventional system performance indicators. Feed channel pressure drop and total cell concentration in the Membrane Fouling Simulator (MFS) flowcell cross-flow outlet water were monitored over time as early biofouling indicators. The results demonstrated the potential of increased bacterial cell concentration in cross-flow outlet water as a reliable indicator of biofouling development on the membrane. Water outlet monitoring enabled faster biofouling detection compared to feed channel pressure drop. Membrane autopsy confirmed biofilm presence prior to the pressure drop increase, highlighting the advantage of early detection in implementing corrective measures. Timely intervention reduces operational costs and energy consumption in membrane-based processes.

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.

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.

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, 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.

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.

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.

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.

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.

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.