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.

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.

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.

The use of traditional drinking water microbial quality monitoring methods, including heterotrophic plate counts (HPCs) and total coliform counts, are not only laborious and time-consuming but also do not readily allow identification of risk areas in the network. Furthermore, if areas of concern are identified, and mitigation measures are taken, it takes days before the effectiveness of these measures is known. This study identified flow cytometry (FCM) as an online sensor technology for bacterial water quality monitoring in the distribution network. We monitored the total bacterial cell numbers and biodiversity in a drinking water distribution system (DWDS) using an online FCM. Two parallel online FCM monitoring systems were installed on two different locations at a drinking water treatment plant (DWTP; Saudi Arabia) supplying chlorinated water to the distribution and in the network 3.6 km away from the DWTP. The FCMs were operated at the same time in parallel to assess the biological stability in DWDSs. The flow cytometric data was compared with the conventional water quality detection methods (HPC and total coliforms). HPC and total coliforms were constantly below the detection limits, while the FCM provided detectable total cell count data and enabled the quantification of changes in the drinking water both with time and during distribution. Results demonstrate the value of FCM as a tool for compliance monitoring and risk assessment of DWDSs.

This study aimed to characterize the impact of organic and inorganic compounds in the feed solution on the membrane fouling and the boron flux in a forward osmosis (FO) membrane system. Lab-scale FO membrane systems were operated in batch mode, with a solution containing organic compounds (humic acid (HA), bovine serum albumin (BSA), sodium alginate (SA)), scaling constituents (calcium chloride, sodium sulfate), and boric acid as feed solution, and with concentrated sodium chloride as draw solution. Preparations of membrane sample for scanning electron microscopy and energy dispersed spectroscopy (SEM-EDX) by freeze- and by oven-drying showed different results and can therefore not be considered as suitable for the evaluation of the samples. Results showed that the organic fouling layer did not have an impact on water and boron fluxes, but reverse salt flux (RSF) was reduced in the presence of BSA and HA. The molar flux ratio between water and boron was roughly 10⁵, regardless of the presence of the fouling. The hydrophobicity and surface charge of the fouling layer were not related to the boron transport rate. A thicker and more uniform organic fouling layer, as observed, caused a reduced RSF. The organic compounds in the feed solution impeded the crystallisation of gypsum, which can lead to reduce scaling.

Phosphate limitation has been suggested as a preventive method against biofilms. P-limited feed water was studied as a preventive strategy against biofouling in cooling towers (CTs). Three pilot-scale open recirculating CTs were operated in parallel for five weeks. RO permeate was fed to the CTs (1) without supplementation (reference), (2) with supplementation by biodegradable carbon (P-limited) and (3) with supplementation of all nutrients (non-P-limited). The P-limited water contained ≤10 µg PO₄ l⁻¹. Investigating the CT-basins and coupons showed that P-limited water (1) did not prevent biofilm formation and (2) resulted in a higher volume of organic matter per unit of active biomass compared with the other CTs. Exposure to external conditions and cycle of concentration were likely factors that allowed a P concentration sufficient to cause extensive biofouling despite being the limiting compound. In conclusion, phosphate limitation in cooling water is not a suitable strategy for CT biofouling control.

We investigated the impact of conditioning compositions on the way bacteria move and adhere to reverse osmosis (RO) membranes that have been pre-conditioned by organic compounds. We used humic acid (HA), bovine serum albumin (BSA), and sodium alginate (SA) to simulate conditioning layers on the RO membranes. First, we investigated the chemotactic responses of Pseudomonas aeruginosa PAO1 to the organic substances and the impact of changes in physicochemical characteristics of pre-conditioned membranes on bacterial attachment. Second, we observed bacterial attachment under the presence or absence of nutrients or microbial metabolic activity. Results showed that there was no relationship between the chemotactic response of P. aeruginosa PAO1 and the organic substances, and the changes in hydrophobicity, surface free energy, and surface charge resulting from changing the composition of the conditioning layer did not seem to affect bacterial attachment, whereas changing the roughness of the conditioned membrane exponentially did (exponential correlation coefficient, R² = 0.85). We found that the initial bacterial attachment on the membrane surface is influenced by (i) the nutrients in the feed solution and (ii) the microbial metabolic activity, whereas the chemotaxis response has a negligible impact. This study would help to establish a suitable strategy to manage bacterial attachment.

The growing demand for fresh water has resulted in increasing use of reverse osmosis (RO) membrane systems for seawater desalination. A major operational problem of RO membrane filtration systems is biofouling – biomass growth causing an unacceptable membrane performance decline. Biofouling reduces the produced water quantity and quality and increases costs. The need for fouling remediation is mainly derived from destructive trial-and-error research with practical membrane modules. Therefore, a clear need existed for the development of a small-sized membrane fouling simulator (MFS) for systematic, low-cost studies. Since the introduction of the MFS in 2006, many articles have appeared which are evaluated in this review. The review describes (i) the location of biofouling in full-scale installations, (ii) development of MFS, (iii) characterization, reproducibility and representativeness of fouling development in the MFS, (iv) applications such as assessing the impact of anti-scalant or biocide dosage, phosphate limitation, feed spacer geometry and linear flow velocity and (v) early warning for biofouling. MFS studies have increased the understanding of biofouling and enabled improved practical membrane performance such as selection of dosed chemicals and feed spacer design. Future MFS studies are anticipated to enable the development of advanced biofouling control strategies.