Search

Desalination brine is a hypersaline byproduct that contains various operational chemicals such as polyphosphonate-based antiscalants. Brine often sinks and flows over the seabed by density currents; therefore, it may affect sediment-water nutrient fluxes and thus microbial activity. We quantified these parameters in brine plumes around two large-scale desalination facilities located in the P-limited Southeastern Mediterranean Sea. The benthic nutrient fluxes and microbial activity were determined using

Many coral reefs are found in arid and semi-arid regions that often face severe water scarcity and depend on seawater desalination for freshwater supply. Alongside freshwater production, desalination plants discharge brine waste into the sea. Brine includes various chemicals (e.g., antiscalants) that may harm the coastal environment. Although widely used, little is known about the ecotoxicological effects of antiscalants (AS) on hard corals. This study compared the impacts of polyphosphonate-based and polymer-based ASs on the coral Montipora capricornis. After two weeks of exposure, we determined the effects of AS on coral physiology, symbiotic microalgae, and associated bacteria, using various analytical approaches such as optical coherence tomography, pulse amplitude modulated fluorometry, and oxidative stress biomarkers. Both ASs reduced polyp activity (∼25%) and caused tissue damage (30% and 41% for polymer and polyphosphonate based AS, respectively). In addition, exposure to polyphosphonate-based AS decreased the abundance of endosymbiotic algae (39%) and upregulated the antioxidant capacity of the animal host (45%). The microalgal symbionts were under oxidative stress, with increased levels of antioxidant capacity and oxidative damage (a 2-fold increase compared to the control). Interestingly, exposure to AS enhanced the numbers of associated bacteria (∼40% compared to the control seawater) regardless of the AS type. Our results introduce new insights into the effects of brine on the physiology of hard corals, highlighting that choosing AS type must be examined according to the receiving ecosystem.

Biofouling in anaerobic membrane bioreactors (AnMBRs) has not been studied widely. Moreover, the effect of membrane surface properties on biofilm formation beyond initial deposition is controversial. We investigated biofouling with polyvinyldifluoride, polyacrylonitrile, and zwitterion-modified polyethersulfone ultrafiltration membranes having different properties during 72 h filtration using natural anaerobes isolated from AnMBR and analyzed biofilm characteristics by physicochemical and molecular techniques. A decrease in membrane performance was positively correlated with biofilm formation on polyvinyldifluoride and polyacrylonitrile membranes, and as expected, physical cleaning effectively mitigated biofilm on hydrophilic and low-roughness membranes. Surprisingly, while the biofilm on the hydrophilic and low-surface roughness zwitterion-modified membrane was significantly impaired, the impact on transmembrane pressure was the highest. This was ascribed to the formation of a soft compressible thin biofilm with high hydraulic resistance, and internal clogging and pore blocking due to high pore-size distribution. Anaerobe community analysis demonstrated some selection between the bulk and biofilm anaerobes and differences in the relative abundance of the dominant anaerobes among the membranes. However, correlation analyses revealed that all membrane properties studied affected microbial communities' composition, highlighting the system's complexity. Overall, our findings indicate that the membrane properties can affect biofilm formation and the anaerobic microbial population but not necessarily alleviate biofouling.

Municipal sewage carries degraded and intact viral particles and RNA (ribonucleic acid) of SARS-CoV-2 (severe acute respiratory coronavirus 2), shed by COVID-19 (coronavirus disease 2019) patients, to sewage and eventually to wastewater treatment plants. Proper wastewater treatment can prevent uncontrolled discharges of the virus into the environment. However, the role of different wastewater treatment stages in reducing viral RNA concentrations is, thus far, unknown. Here, we quantified SARS-CoV-2 RNA in raw sewage and during the main stages of the activated sludge process from two wastewater treatment plants in Israel, on three different days during the 2020 COVID-19 outbreak. To reduce the detection limit, samples were concentrated prior to quantification by real-time polymerase chain reaction by a factor of 2–43 using ultrafiltration. On average, ∼1 log RNA removal was attained by each of the primary and secondary treatment steps; however, >100 copies of SARS-CoV-2 RNA/mL remained in the secondary effluents. Following chlorination, SARS-CoV-2 RNA was detected only once, likely due to an insufficient chlorine dose. Our results emphasize the capabilities and limitations of the conventional wastewater treatment process in reducing the SARS-CoV-2 RNA concentration and present preliminary evidence for the importance of tertiary treatment and chlorination in reducing dissemination of the virus to the environment., We present data on SARS-CoV-2 RNA concentration and removal rates during the different stages of the activated sludge wastewater treatment process with implications for controlling environmental viral dissemination.

The bacteriophage infection cycle has been extensively studied, yet little is known about the nanostructure and mechanical changes that lead to bacterial lysis. Here, atomic force microscopy was used to study in real time and in situ the impact of the canonical phage T4 on the nanotopography and biomechanics of irreversibly attached, biofilm-forming E. coli cells. The results show that in contrast to the lytic cycle in planktonic cells, which ends explosively, anchored cells that are in the process of forming a biofilm undergo a more gradual lysis, developing distinct nanoscale lesions (~300 nm in diameter) within the cell envelope. Furthermore, it is shown that the envelope rigidity and cell elasticity decrease (>50% and >40%, respectively) following T4 infection, a process likely linked to changes in the nanostructure of infected cells. These insights show that the well-established lytic pathway of planktonic cells may be significantly different from that of biofilm-forming cells. Elucidating the lysis paradigm of these cells may advance biofilm removal and phage therapeutics.

Initial deposition of bacteria is a critical stage during biofilm formation and biofouling development in membrane systems used in the water industry. However, the effects of hydrodynamic conditions on spatiotemporal deposition patterns of bacteria during the initial stages of biofilm formation remain unclear. Large field epifluorescence microscopy enabled in situ and real-time tracking of Bacillus subtilis in a forward osmosis system with spacers during the first 4 h of biofilm formation. This study quantitatively compares the spatiotemporal deposition patterns between different hydrodynamic conditions: high and low permeate water flux (6 or 30 L m-2 h-1) as well as high and low crossflow velocity (1 or 14 cm s-1). Low crossflow velocity and high permeate water flux maximized bacterial attachment to the membrane surface, which was 60 times greater (6 × 103 cells mm-2) than at high crossflow velocity and low permeate water flux (

Circumstantial evidence has suggested that jellyfish swarms impair the operation of seawater reverse osmosis desalination facilities. However, only limited information is currently available on the pretreatment efficiency of jellyfish and their effects on reverse osmosis (RO) membrane performance. Here, we have comprehensively tested the pretreatment efficiency of a dual-media gravity filter and cartridge micro-filtration following the addition of jellyfish into the feedwater. Concurrently, the fouling propensity and performance of the RO membranes were examined. We show that jellyfish demise resulted in seawater eutrophication that triggered a significant increase in bacterial biomass (∼50-fold), activity (∼7-fold), and release of transparent exopolymer particles (∼5-fold), peaking three days after the addition of jellyfish into the feedwater. In parallel, a significant reduction in permeate water flux was recorded (∼10%) while trans-membrane pressure sharply increased (15%), reaching the operation pressure limit of our system (75 bar) after five days. At the conclusion of the experiments, the membrane surface was heavily covered by large chunks of organic-rich material and multilayered biofilms. Our results provide a holistic view on the operational challenges of seawater reverse osmosis (SWRO) desalination triggered by jellyfish swarms in coastal areas. Following the above, it can be inferred that freshwater production will likely be halted three days after drawing the jellyfish into the pretreatment system. Outcomes from these results may lead to the development of science-based operational protocols to cope with growing occurrence of jellyfish swarms around the intake of SWRO desalination facilities worldwide.

Recent studies have indicated that heterotrophic diazotrophs are highly diverse and fix N

Sewage outbursts affect coastal environments as seawater is enriched with nutrients, organic matter and microbes, thus can potentially impair seawater reverse osmosis (SWRO) desalination. In this study, we evaluated how municipal sewage outbursts affect SWRO desalination in a pilot-scale system. To this end, feedwater characteristics (i.e., coastal water), the removal efficiency of organic foulants by a dual-media gravity filter, and cartridge micro-filtration were determined daily for 12 days. Permeate water flux was maintained constant during the study, while trans-membrane pressure (TMP) was automatically adjusted and continuously monitored. The results indicate that sewage outbursts caused an immediate (∼1 d) buildup of phyto/bacterioplankton biomass (up to 10-fold), and enhanced activity (maximal 30-fold) followed by an increase in transparent exopolymer particle (TEP) concentrations. After sewage addition, algal biomass was significantly removed by the pretreatment system (72–90%), while a considerable fraction of the bacterial biomass (42–65%) and TEP (53–65%) passed these procedures. The result was a negative impact on the desalination performance reflected by a significant increase (> 10%) in RO-TMP 7.5 d after the sewage addition. Our results indicate on a direct link between sewage outbursts, pretreatment efficiency, and SWRO desalination. Nevertheless, these findings can lead to new avenues for the development of science-based operational protocols to minimize the deleterious effects of abrupt sewage outbursts on SWRO desalination.

Human-induced eutrophication of coastal water may be a major threat to aquatic life. Here, we investigated the effects of N-rich well amelioration brines (WAB) on coastal phytoplankton population's habitat in the surface oligotrophic waters of the southeastern Mediterranean Sea (SEM). To this end, we added WAB (2 concentrations) to mesocosms (1-m3 bags) to surface SEM water during summer and winter, where changes in phytoplankton biomass, activity and diversity was monitored daily for 8 days. Our results demonstrate that WAB addition triggered a phytoplankton bloom, resulting in elevated algal biomass (maximal +780%), increased primary production rates (maximal +675%) and a decrease in eukaryotic algal α-diversity (ca. −20%). Among the species that bloomed following WAB amendments, we found the potentially toxic dinoflagellate Karlodinium venificum. This study adds valuable perspective to the effect of nutrients discharged into nutrient limited SEM coastal waters, and in particular of N-derived WAB.

Seawater desalination facilities continuously discharge hyper-saline brine into the coastal environment which often flows as a concentrated plume over the seafloor, hence possibly impacting benthic microorganisms. Yet, the effects of brine discharge from desalination plants on benthic bacteria, key players in biodegradation of organic material and nutrient recycling is unknown. In this study, we tested the chronic (years) effects of brine discharge from three large-scale desalination facilities on the abundance, metabolic activity and community composition of benthic bacteria. To this end, four sampling campaigns were carried at the outfall areas of the Ashkelon, Sorek and Hadera desalination facilities. The effects of the brine were compared to corresponding reference stations which were not influenced by the brine (i.e., water temperature and salinity). Our sampling data indicate that bacterial abundance and activity that includes bacterial growth efficiency were 1.3–2.6-fold higher at the outfall area than the reference station. Concomitant analysis pointed out that the bacterial community structure at the brine discharge area was also different than the reference station, yet varied between each desalination facility. Our results demonstrate that the impact of brine effluent from desalination facilities on benthic bacteria are site-specific and localized (

Bacteriophages, or phages, are receiving increasing interest as recognition tools for the design of bioactive surfaces. However, to maintain the activity of surface-bound phages, the immobilization strategy must provide the right orientation and not compromise the phages’ integrity. The objectives of this study were to characterize the phage sorption capacity and the immobilized phage activity for aminated silica particles functionalized with T4 phages. Two functionalization strategies were compared; physisorption, based on electrostatic adhesion, and chemisorption, where the phage and the particle are coupled using a carbodiimide cross-linker. We report that chemisorption, at maximum adsorption conditions on 1 µm particles, yielded 16 functional phages per particle, which is 2.5 times more than by the physisorption method. Particle diameter is shown to have an important impact on phage attachment and 1.8 µm particles were found to have ∼4 times more phages per surface area than 0.5 µm particles. Higher surface coverage is attributed to the lower steric hindrance on bigger particles. These findings provide important guidelines for the design of phage-functionalized particles for environmental, biomedical, or sensing applications.

Bacteriophage infection cycle has been extensively studied, yet little is known on the structural and mechanical changes that lead to bacterial lysis. Here, bio-atomic force microscopy was used to study in real-time andin-situthe impact of the canonical phage T4 on the nano-topography and biomechanics of irreversibly attached, biofilm-formingE. colicells. The results show that in contrast to the lytic cycle in planktonic cells, which ends explosively, anchored cells that are in the process of forming biofilms undergo gradual lysis, developing distinct sub-micron lesions (∼300 nm in diameter) within the cell envelope. Furthermore, it is shown that the envelope rigidity and cell elasticity decrease (>50% and >40%, respectively) following T4 infection. These new insights show that the well-established lytic pathway of planktonic cells may be significantly different from that of biofilm-forming cells. Elucidating the lysis paradigm of these cells may advance biofilm removal and phage therapeutic.There is no conflict of interest and all co-authors have seen and approved the current version for submission.

Planktonic heterotrophic diazotrophs (N2-fixers) are widely distributed in marine and freshwater systems, yet limited information is available on their activity, especially in environments with adverse conditions for diazotrophy (e.g., N-rich and oxygenated). Here, we followed the localization and activity of heterotrophic diazotrophs in the hyper-eutrophic N-rich Qishon River-an environment previously considered to be unfavorable for diazotrophy. Our results indicate high heterotrophic N2 fixation rates (up to 6.9 nmol N L-1 d-1), which were approximately three fold higher at an upstream location (freshwater) compared to an estuary (brackish) site. Further, active heterotrophic diazotrophs were capture associated with free-floating aggregates by a newly developed immunolocalization approach. These findings provide new insights on the activity of heterotrophic diazotrophs on aggregates in environments previously considered with adverse conditions for diazotrophy. Moreover, these new insights may be applicable to other aquatic regimes worldwide with similar N-rich/oxygenated conditions that should potentially inhibit N2 fixation.

The marine cyanobacterium Prochlorococcus is a dominant photoautotroph in many oligotrophic Low-Nutrients-Low-Chlorophyll (LNLC) regions. While the chemical impact of aerosols upon interaction with surface seawater was documented in numerous studies, we show that Prochlorococcus cells are affected also by bio-aerosols (potentially biological agents in the dust/aerosols such as membrane-bound extracellular vesicles, small-size bacteria and/or viruses), resulting in lower surface seawater abundances in the oligotrophic Mediterranean Sea. We conducted experimental amendments of &lsquo, live&rsquo, aerosol/dust particles and aerosol filtrates (&lt, 0.22-&micro, m) to surface Southeastern Mediterranean seawater or to pure Prochlorococcus cultures (MED4). Results show a significant decline in cell biomass (&lt, 90%), while UV-sterilized aerosols elicited a much weaker and non-significant response (~10%). We suggest that the difference is due to a negative effect of bio-aerosols specific to Prochlorococcus. Accordingly, the dominance of Synechococcus over Prochlorococcus throughout the surface Mediterranean Sea (observed mainly in spring when atmospheric aerosol levels are relatively high) and the lack of spatial westward gradient in Prochlorococcus biomass as typically observed for chlorophyll-a or other cyanobacteria may be attributed, at least to some extent, to the impact of bio-aerosol deposition across the basin. Predictions for enhanced desertification and increased dust emissions may intensify the transport and potential impact of bio-aerosols in LNLC marine systems.

Membrane distillation (MD) is a temperature driven membrane separation technology that holds great potential for decentralized and sustainable wastewater treatment systems. Yet, similarly to all membrane based systems, microbial fouling (biofouling) might be a critical hurdle for MD wastewater treatment applications. In this study we determined the impact of increasing feedwater temperatures (47 °C, 55 °C, and 65 °C) on biofilm growth and MD performance via dynamic biofouling experiments with Anoxybacillus sp. as a model bacterium. Our results indicated that cell growth was reduced at 47 °C, resulting in moderate distillate water flux decline (30%). Differently, extensive growth of Anoxybacillus sp. at feedwater temperature of 55 °C caused severe distillate water flux decline (78%). Additionally, biofouling induced membrane wetting, which facilitated the passage of bacteria cells and endospores through the membrane structure into the distillate. Although bacterial growth was impaired at feedwater temperatures of 65 °C, excessive production of EPS (compared to bacterial abundance) crippled membrane separation due to severe pore wetting. These results underline the importance of optimized operating conditions and development of antibiofouling and antiwetting membranes for successful implementation of MD in wastewater treatment with high biofouling propensity.

Forward osmosis (FO) has gained increasing attention in desalination, wastewater treatment, and power generation. However, biofouling remains a major obstacle for the sustainable development of the FO process. Both passive and active strategies have been developed to mitigate membrane biofouling. A comprehensive understanding of different strategies and mechanisms has fundamental significance for the antifouling membrane development. In this study, thin-film composite (TFC) FO membranes were modified with polydopamine (PDA) coating as a passive antibacterial moiety and silver nanoparticles (Ag NPs) as an active antibacterial moiety. Their anti-biofouling performances were investigated both in static and dynamic conditions. In static exposure, the PDA-coated membranes exhibited great passive anti-adhesive property, and the Ag-NP-generated membranes presented both of excellent passive anti-adhesive properties and active antibacterial performance. While in dynamic cross-flow running conditions, Ag NPs effectively mitigated the membrane water flux decline due to their inhibition of biofilm growth, the PDA coating failed because of its inability to inactivate the attached bacteria growth. Moreover, Ag NPs were stable and active on membrane surfaces after 24 h of cross-flow operation. These findings provide new insights into the performances and mechanisms of passive and active moieties in the FO process.

Fouling continues to be a limiting factor in membrane processes. Pulsating feed flows have the potential to decrease fouling while maintaining a constant pressure drop over the module. This study quantifies the impact of pulsating flows on particle deposition in a forward osmosis system with spacers at a mean crossflow velocity of 14 cm s−1 and permeate water flux of 18 L m−2 h−1. Carboxylate modified polystyrene beads were tracked by fluorescence microscopy in situ and real-time during the first 4 h of deposition with feed flow pulsations up to a frequency of 10 Hz, while experiments were conducted with a constant crossflow velocity as a reference. Imaging at 30 s intervals revealed deposition rates of up to 2.6 times higher at steady state, as compared to flow pulsations of 10 Hz. Consequentially, average deposition was reduced by a factor of 3.5 from steady state (109 ± 67 beads mm−2) to pulsating feed flow (31 ± 17 beads mm−2) after 4 h of deposition. The results of this research provide new fundamental insights related to the constraints and advantages of using pulsating flows for fouling mitigation in forward osmosis systems.

The water-energy nexus has motivated the quest for new membrane-based technologies that target potable water and energy production. To this end, membrane distillation (MD), forward osmosis (FO) and pressure retarded osmosis (PRO) provide alternative means for the sustainable production of freshwater and electricity from feed water with high fouling potential such as wastewater. MD is a thermally driven process that can utilize low grade (latent) heat sources, while FO and PRO harness osmotic gradients as the driving force. High rejection of contaminants, compact modular design and low fouling propensity make these membrane technologies suitable for treating different types of wastewater. However, the application of feed solutions with high loads of organic matter and bacteria prompts the development of microbial fouling (biofouling), which significantly reduces system performance. Therefore, mitigating biofouling by minimizing bacterial attachment and enhancing the biofilm cleaning efficiency is imperative. We stress that in-depth exploration of the impacts imposed by biofilm in MD, FO and PRO systems is essential before developing new approaches for biofouling mitigation. This comprehensive review compiles the driving forces of these non-pressurized membrane systems, while focusing on the current knowledge regarding the various impacts of biofouling. Moreover, we highlight current and future research directions that focus on the development of new approaches to minimize MD, FO and PRO biofouling.

Seawater reverse osmosis (SWRO) desalination discharge brine-waste as part of the process with immediate (short-term) and chronic (long-term) impacts to marine coastal environment. Brine-waste is often denser than the receiving environment, therefore sinks and flows as a saline plume in adjacency to the sea floor. We suggest that the saline flow over the bottom may impose an osmotic stress, and alter benthic heterotrophic microbial diversity, activity and growth. In this study, we examined the short-term (48 h) effects of different salinities on benthic heterotrophic bacteria attached to the sediment at the eastern Mediterranean coast. To this end, 12 cylinders were filled with sediment and incubated in full darkness with rising salinities ranging from 2% to 20% over the ambient levels. During the summer experiments, heterotrophic bacterial abundance has reduced by 60% at salinity levels > 5% above the ambient concentration. Further, bacterial cell specific activity significantly increased following high salinity scenarios. Our experimental results provide the first scientifically-based data on the immediate effects of SWRO brine over benthic heterotrophic bacteria. However, we stress that long-term studies are imperative at the outfall of operating desalination facilities to determine the chronic effects of brine on benthic bacteria.

N2 fixation by planktonic heterotrophic diazotrophs is more wide spread than previously thought, including environments considered “unfavorable” for diazotrophy. These environments include a substantial fraction of the aquatic biosphere such as eutrophic estuaries with high ambient nitrogen concentrations and oxidized aphotic water. Different studies suggested that heterotrophic diazotrophs associated with aggregates may promote N2 fixation in such environments. However, this association was never validated directly and relies mainly on indirect relationships and different statistical approaches. Here, we identified, for the first time, a direct link between active heterotrophic diazotrophs and aggregates that comprise polysaccharides. Our new staining method combines fluorescent tagging of active diazotrophs by nitrogenase-immunolabeling, polysaccharides staining by Alcian blue or concanavalin-A, and total bacteria via nucleic-acid staining. Concomitant to N2 fixation rates and bacterial activity, this new method provided specific localization of heterotrophic diazotrophs on artificial and natural aggregates. We postulate that the insights gained by this new visualization approach will have a broad significance for future research on the aquatic nitrogen cycle, including environments in which diazotrophy has traditionally been overlooked.

Bacteria of different Gram-types have inherently different outer cell structures, influencing cell surface properties and bacterial attachment. Dynamic biofouling experiments were conducted over four days in a bench-scale forward osmosis (FO) system with Gram-negative Pseudomonas aeruginosa or Gram-positive Anoxybacillus sp. Biofouling resulted in ∼10% decline in FO permeate water flux and was found to be significant for Anoxybacillus sp. but not for P. aeruginosa. Additionally, a stronger permeate water flux decline for P. aeruginosa in experiments with a superhydrophilic feed spacer demonstrated that mitigation methods require testing with different bacterial Gram-types. It was found that although permeate water flux decline can be affected by bacterial Gram-type the stable performance under enhanced biofouling conditions highlights the potential of FO for wastewater reclamation.

Biofouling impacts seawater reverse osmosis (SWRO) desalination plants by hindering module performance, increasing energetic demands, and incurring further costs. Here we investigated the spatial–temporal dynamics of microbial communities along the feedwater, pretreatment, and reverse osmosis stages of a large-scale SWRO desalination facility. While the composition of water-based microbial communities varied seasonally, the composition of biofilm microbial communities clustered by locations. Proteobacteria dominated throughout the water and biofilm communities while other dominant phyla varied seasonally and spatially. The microbial community composition significantly differed along the pathway locations of feedwater, rapid sand filtration (RSF), cartridge filters (CF), and the reverse osmosis (RO) membranes. Biofilms on the RSF and CF were composed of more diverse microbial populations than RO biofilms as determined by the effective number of species. Biofilms that developed along the treatment pathway (CF) served as inocula enhancing biofouling downstream on the RO membranes. Subsequently, we believe that prior to the development of advanced antibiofouling treatments for the desalination industries, the site-specific microbial community of feedwater, pretreatment and RO biofouling should be characterized. Site specific identification of these communities will enable optimization of pretreatment and cleaning procedures and can ultimately reduce chemical usage and incurred costs.

Fouling is the main hurdle for all membrane-based systems treating water with high fouling propensity. A novel imaging approach was applied in a forward osmosis (FO) system with spacers to follow in-situ and real-time deposition of fluorescent beads as bacteria proxy on the membrane using large-field high-resolution epifluorescence microscopy. For the first time, this study quantifies the impact of crossflow velocity (1.3 or 13 cm s-1) and permeate water flux (6 or 30 L m-2 h-1) on spatiotemporal patterns of initial cake layer formation (4 h). The total amount of deposited particles on the membrane increased by 84 fold as the ratio of permeate water flux over crossflow velocity, D, was raised by 54 fold. Spatial distribution of particles was more homogenous at higher D ratio, while particle accumulation rates decreased by 50% over 4 h. Distribution of local velocities of particle flow paths elucidated the observed spatial deposition patterns. These new quantitative results highlight that the ratio of permeate water flux over crossflow velocity impacts all aspects of particle deposition and may aid in designing new spacer geometries. We also suggest that appropriate hydrodynamic conditions may be a viable tool to postpone the onset of fouling in new and cleaned membrane modules.

Deposition and flow paths of particles (live and inert) often control the first stages of “cake” (fouling) formation in membrane systems. Developments in crossflow cell design such as three-dimensional (3D) printing and state of the art imaging techniques enable real-time and in situ tracking of initial cake formation. In this work, high-resolution large-field fluorescent microscopy was used to concomitantly study and compare the deposition and flow paths of inert beads and Bacillus subtilis as biofilm forming bacteria. The membrane surface was imaged continuously (30 s intervals) during the initial stages of cake formation (

This study investigates the biofouling resistance of modified reverse osmosis (RO) feed spacers. Control spacers (made of polypropylene) were functionalized with a biocidal coating (silver), hydrophilic (SiO2 nanoparticles) or superhydrophobic (TMPSi-TiO2 nanoparticles) anti-adhesive coatings, or a hybrid hydrophilic-biocidal coating (graphene oxide). Performance was measured by adhesion assays, viability tests, and permeate flow decline in a bench scale RO system. The control spacers proved to be one of the better performing materials based on bacterial deposition and dynamic RO fouling experiments. The good anti-adhesive properties of the control can be explained by its near ideal surface free energy (SFE). The only surface modification that significantly reduced biofouling compared to the control was the biocidal silver coating, which outperformed the other spacers by all measured indicators. Therefore, future efforts to improve spacer materials for biofouling control should focus on engineering biocidal coatings, rather than anti-adhesive ones.

Author(s): Petersen, Karen Lykkebo | Advisor(s): Paytan, Adina | Abstract: Terrestrial water resources are scarce in arid and semi-arid regions of the world and increasing demands for water worldwide are adding additional pressures on limited water resources. Seawater desalination provides a reliable source of potable water. The process of desalination creates a high-salinity byproduct that is discharged back into the coastal environment by various methods (pipes, diffusers, channels). The brine effluent is often mixed with chemicals used at the desalination facilities and can, without dilution; reach salinities twice of the ambient feed-water. Despite a growing desalination capacity worldwide, the effects of that brine discharge on local coastal environments are not fully understood or constrained. This thesis investigates the impacts of desalination brine discharge on coastal biology and chemistry, and is comprised of three chapters: Chapter 1 investigates the in-situ impacts of desalination discharge measured before and after the Carlsbad Desalination Plant (California) became operational on both water chemical parameters and benthic macro fauna; Chapter 2 investigates the impacts of desalination brine on corals from The Gulf of Aqaba in a 4-week incubation experiment with salinities 10% above ambient and addition of phosphonate-based antiscalants; and Chapter 3 reviews current knowledge of the impacts of brine discharge on a wide range of coastal organisms.The Carlsbad Desalination Plant became operational in December 2015, and water, sediment and biological samples were collected prior to operations (December 2014 and September 2015) and after operations began (May and November 2016). A distinct brine plume of salinity 34-37 (ambient salinity 33.3) extending 600m offshore from the discharge point was observed after the plant started discharging. This is despite the coastal area being “high-energy” and previous computer models predicting mixing of the brine to be better. The salinity plume does not have an immediate effect on the benthic environment: Macro fauna abundance (tube-forming Polychaetes) significantly increases compared to post-operation surveys, however the in-faunal organisms show no change. The benthic area around the outfall has same level of disturbance pre and post operations, possibly because of the continuous discharge from the local power plant. A 5-week bioassay of brittle stars in brine treatments indicated a trend towards decreased growth and impaired agility in increased salinities but no significant change was observed. Finally, using mean wave height along the California coast as a proxy for coastal mixing, locations of proposed desalination facilities are compared to the location of Carlsbad Desalination Plant to evaluate the expected mixing properties of those areas. In the brine incubation experiment of hard corals (Stylophora pistillata, Acropora tenuis, and Pocillopora verrucosa) there were an overall decrease in coral performance and changes in the corals’ physiology. Symbiodinium abundance, protein content and heterotrophic abundances were significantly reduced at the end of the incubation in all three species. In particular, the addition of phosphonate-based antiscalant to a 10% salinity increase reduced the amount of corals tissue symbionts, calcification rates and oxygen production rates. These results suggest that coral reefs may be susceptible to exposure to SWRO discharge and calls for careful selection of locations of SWRO facilities around coral reefs.Various marine species have various response and resilience to increased salinity and brine discharge. To increase SWRO facilities sustainability and minimize their footprint in the coastal environment, sufficient mixing of the brine effluent is important. Diffusor systems generally provide the best mixing, but co-location of SWRO facilities with power plants, already discharging cooling water, minimizes additional impacts of the brine. Local monitoring of the marine benthic environment is encouraged in locations of proposed desalination facilities.

The southeastern Mediterranean Sea (SEMS) is a warm and sunlit marine environment with low ambient N concentration, thus considered ideal for diazotrophy by autotrophic diazotrophs such as Trichodesmium. Despite the favorable conditions, N2 fixation rates are often low and Trichodesmium has hardly been spotted in the SEMS. This study reports on the occurrence of a Trichodesmium bloom in the SEMS which was ascribed to T. erythraeum according to DNA fingerprinting of the nifH gene. We found that this bloom (1407 ± 983 cells L−1) was triggered by an intense outburst of raw sewage that supplied high concentrations of N, P and dissolved organic carbon (DOC), which resulted in low N:P (~12:1) and exceptionally high C:P (~1340:1) ratios. We surmise that these conditions provided favorable conditions for Trichodesmium bloom to form via mixotrophic metabolism. As a result, a fourfold increase in N2 fixation was recorded, which contributed ~70% to new primary production and spur a sharp increase in phytoplankton activity and biomass. The conclusions of this study point on a new paradigm for bloom-forming T. erythraeum which is tightly linked to anthropogenic sources and prompt microbial productivity in oligotrophic marine environments such as the SEMS.

Membrane distillation (MD) is an emerging desalination technology that uses low-grade heat to drive water vapor across a microporous hydrophobic membrane. Currently, little is known about the biofilms that grow on MD membranes. In this study, we use estuarine water collected from Long Island Sound in a bench-scale direct contact MD system to investigate the initial stages of biofilm formation. For comparison, we studied biofilm formation in a bench-scale reverse osmosis (RO) system using the same feedwater. These two membrane desalination systems expose the natural microbial community to vastly different environmental conditions: high temperatures with no hydraulic pressure in MD and low temperature with hydraulic pressure in RO. Over the course of 4 days, we observed a steady decline in bacteria concentration (nearly 2 orders of magnitude) in the MD feed reservoir. Even with this drop in planktonic bacteria, significant biofilm formation was observed. Biofilm morphologies on MD and RO membranes were markedly different. MD membrane biofilms were heterogeneous and contained several colonies, while RO membrane biofilms, although thicker, were a homogeneous mat. Phylogenetic analysis using next-generation sequencing of 16S rDNA showed significant shifts in the microbial communities. Bacteria representing the orders Burkholderiales, Rhodobacterales, and Flavobacteriales were most abundant in the MD biofilms. On the basis of the results, we propose two different regimes for microbial community shifts and biofilm development in RO and MD systems.

Confocal laser scanning microscopy (CLSM) is often used to evaluate biofilm development or biofouling mitigation in membrane systems. However, several methods of CLSM sample preparation exist. In this paper, we evaluate the effects of three preparation techniques — dry, confined (wet), and unconfined (immersed) mounting — on CLSM-derived biofilm architecture and dimensions. Although placing a wet or dry biofilm between a slide and a coverslip before viewing is relatively common, our results show that this confinement significantly alters the biofilm observed. Therefore, biofilms should be viewed in an unconfined and hydrated state that allows for full extension of the biofilm structure in a media-filled viewing well of fixed depth (~ 250 μm). Pseudomonas aeruginosa biofilms were grown on thin-film composite reverse osmosis membranes and glass coupons. Dry and confined mounting of 24 and 48 h biofilms resulted in biofilms with low 3-D complexity and thickness (14 and 18 μm, respectively). Measured biofilm thickness was significantly higher on samples prepared using unconfined mounting (55 μm). Additionally, the reduction in biofilm thickness and biovolume observed after treatment with biocidal compounds was significantly less on the dry and confined biofilms than the unconfined samples. Our results strongly suggest that biofilms on membranes be prepared for microscopy using unconfined mounting to accurately assess biofilm structure and dimensions. Unconfined mounting will allow for accurate CLSM assessment of membrane biofilm structure, dimensions, and biofouling mitigation measures in membrane systems.

Rapid sand filtration (RSF), proceeded by chemical coagulation and flocculation, is a commonly used, effective pretreatment in the desalination industry. We designed and tested a novel, large pilot-scale, two-stage granular Rapid Bioflocculation Filter (RBF) based on a first-stage Bioflocculator (BF) unit followed by a mixed-media bed filter (MBF). The BF filter bed consisted of an extremely porous volcanic Tuff granular medium which provided an enlarged surface area for microbial development and biofilm proliferation. We compared the efficiency of the pilot RBF to that of a full-scale RSF, operating with upstream chemical coagulation, by measuring the removal from the same untreated seawater feed of key factors related to membrane clogging: SDI, turbidity, chlorophyll a (Chl a ) and transparent exopolymer particles (TEP). After 2 weeks of operation, the Tuff grains were colonized extensively by coccoid bacteria that formed biofilm along the entire BF. With bacterial colonization and biofilm development, numerous aggregates of bacteria and some algal cells embedded in an amorphous organic matrix were formed on and within the Tuff grains. By 1–3 months, the biotic diversity within the Tuff filter bed had increased to include filamentous bacteria, cyanobacteria, fungi, protista and even crustaceans and marine worms. During and for ∼24 h after each cleaning cycle (carried out every5 to 7 days by upward flushing with air and water), large numbers of floc-like particles, from ∼15 μm to ∼ 2 mm in size were observed in the filtrate of the BF unit. Microscopic examination of these flocs (stained with Alcian Blue and SYTO R 9) showed that they were aggregates of many smaller particles with associated bacteria and algae within a polysaccharide gel-like matrix. These biogenic flocs ( bioflocs ) were observed to form during normal operation of the RBF, accumulating as aggregates of inorganic and organic material on the Tuff surfaces. With each flush cleaning cycle, these bioflocs were released into the BF effluent but were retained by the second phase MBF unit. No flocs were seen in the MBF filtrate. Over a year-long study, both the pilot RBF and the full-scale RSF showed similar filtration efficiencies, measured as the percentage removal of Chl a , TEP, turbidity and SDI from the same seawater feed. These results indicate the potential of the bioflocculation approach with no chemical additives as an alternative to conventional RSF pretreatment for large SWRO facilities.

[1] In the nitrogen (N)-impoverished photic zones of many oceanic regions, prokaryotic organisms fixing atmospheric dinitrogen (N2; diazotrophs) supply an essential source of new nitrogen and fuel primary production. We measured dinitrogen fixation and primary productivity (PP) during the thermally stratified summer period in different water regimes of the oligotrophic eastern Mediterranean Sea, including the Cyprus Eddy and the Rhodes Gyre. Low N2 fixation rates were measured (0.8–3.2 µmol N m−2 d−1) excluding 10-fold higher rates in the Rhodes Gyre and Cyprus Eddy (~20 µmol N m−2 d−1). The corresponding PP increased from east to west (200–2500 µmol C m−2 d−1), with relatively higher productivity recorded in the Rhodes Gyre and Cyprus Eddy (2150 and 2300 µmol C m−2 d−1, respectively). These measurements demonstrate that N2 fixation in the photic zone of the eastern Mediterranean Sea contributes only negligibly by direct inputs to PP (i.e., cyanobacterial diazotrophs) and is in fact uncoupled from PP. By contrast, N2 fixation is significantly coupled to bacterial productivity and to net heterotrophic areas, suggesting that heterotrophic N2 fixation may in fact be significant in this ultraoligotrophic system. This is further substantiated by the high N2 fixation rates we measured from aphotic depths and by the results of phylogenetic analysis in other studies showing an abundance of heterotrophic diazotrophs.

Transparent exopolymer particles (TEPs) are planktonic, organic microgels that are ubiquitous in aqueous environments. Increasing evidence indicates that TEPs play an active role in the process of aquatic biofilm formation. Frequently, TEPs are intensely colonized by bacteria and other microorganisms, thus serving as hot spots of intense microbial activity. We introduce the term “protobiofilm” to refer to TEPs with extensive microbial outgrowth and colonization. Such particles display most of the characteristics of developing biofilm, with the exception of being attached to a surface. In this study, coastal seawater was passed through custom-designed flow cells that enabled direct observation of TEPs and protobiofilm in the feedwater stream by bright-field and epifluorescence microscopy. Additionally, we could follow biofilm development on immersed surfaces inside the flow cells. Within minutes, we observed TEP and protobiofilm patches adhering to these surfaces. By 30 min, confocal laser-scanning microscopy (CLSM) revealed numerous patches of Con A and SYTO 9 staining structures covering the surfaces. Atomic force microscopy showed details of a thin, highly sticky, organic conditioning layer between these patches. Bright-field and epifluorescence microscopy and CLSM showed that biofilm development (observed until 24 h) was profoundly inhibited in flow cells with seawater prefiltered to remove most large TEPs and protobiofilm. We propose a revised paradigm for aquatic biofilm development that emphasizes the critical role of microgel particles such as TEPs and protobiofilm in facilitating this process. Recognition of the role of planktonic microgels in aquatic biofilm formation can have applied importance for the water industry.

Appropriate devices for obtaining either interstitial water or grain particle samples or for measuring dissolved oxygen (DO) at depth within the interior of large-scale, industrial granular bed filters are currently lacking. We developed two innovative samplers for studying a Rapid Sand Filter (RSF) at an operational SWRO plant: the first for water and granular material, the second for DO measurements. 1. The particle and interstitial water sampler is based on a corer that retains grain particles inside a chamber using water suction created by an external peristaltic pump. 2. The DO sampler incorporates a temperature compensated oxygen measuring optode over which laminar water flow is maintained by an external peristaltic pump. To test these samplers, an operational, full-scale RSF at a SWRO plant was monitored at various depths within the filter bed over several months. Additionally, DO profiles within the filter bed were followed during and after a backwash cycle. These samplers enable efficient and precise collection of DO data, interstitial water, and granular material from previously inaccessible depth layers within the filter bed without interruption of the filter operation, providing information on microbiological processes such as biofilm development, microbial community structure and organic matter cycling.

Rapid sand filtration (RSF) is used today as an effective pretreatment procedure to enhance water quality prior to reverse osmosis (RO) membranes in desalination plants. RSF in newly operated desalination facilities requires a maturation period of about three months before the feedwater may be filtered efficiently. To date the desalination industry RSF has regarded RSF mainly as a physical barrier effectively retaining particles larger than 0.35 mm. In this study we assessed the potential of the RSF as a biological filter by following the dynamics of bacterial colonization and metabolic activity within the filter bed and determining filtration efficiency in respect to particulate and dissolved organic carbon, chlorophyll a and transparent exopolymeric particles (TEP). Bacterial abundance and diversity were shown by DGGE and SEM analysis to increase gradually over a three month sampling period. Bacterial metabolic activity within the filter bed interstitial water increased erratically with time. With the outgrowth of a microbial population and biofilm development on the filter bed medium, significant removal of organic carbon from the source water was always observed. Our results indicate that current RSFs function as biofilters with moderate efficiency. Innovative design is needed to maximize the biofiltration potential of these filters.

N2 fixation should be a critical process in the nitrogen-poor surface water of the eastern Mediterranean Sea. Despite favorable conditions, diazotroph abundance and N2 fixation rates remains low for reasons yet explained. The main goal of this study was to investigate the limiting nutrients for diazotrophy in this oligotrophic environment. Hence, we conducted dedicated bottle-microcosms with eastern Mediterranean Sea water that were supplemented with mono and polysaccharides as well as inorganic nitrogen and phosphorous. Our results indicate that the diazotrophic community expressing nifH was primarily represented by heterotrophic Proteobacteria. N2 fixation and heterotrophic bacterial activity increased up-to tenfold following two days of dark incubations, once seawater was supplemented with organic carbon substrate in the form of glucose (monosaccharides) or gum-xanthan (polysaccharide surrogate). Furthermore, our results point that carbon-rich polysaccharides, such as transparent exopolymer particles, enhance heterotrophic N2 fixation, by forming microenvironments of intense metabolic activity, high carbon: nitrogen ratio, and possibly low O2 levels. The conclusions of this study indicate that diazotrophs in the eastern Mediterranean coast are primarily limited by organic carbon substrates, as possibly in many other marine regions.

Solar light penetrates deep into the clear water of oligotrophic oceans and may have both beneficial and destructive effects on marine phytoplankton. In oligotrophic waters worldwide, phytoplankton communities consist mostly of ultraphytoplankton of the groups Synechococcus and Prochlorococcus, which differ in their optical properties and, therefore, are better suited for thriving in different niches along the natural vertical light gradient. In this article, we aim to draw the optical boundaries separating the two populations in order to get a better insight into the light-driven dynamics in ultraphytoplankton-community structure and to predict future trends. We report spectral, photosynthetically active radiation (PAR) and ultraviolet radiation (UVR) penetration through the stratified season along with temporal and vertical distributions of Synechococcus and Prochlorococcus in the Gulf of Eilat (Aqaba). These light-field parameters are used to define the apparent limits of the vertical distribution of S...

We report N(2) fixation rates measured from two stations monitored monthly off the Mediterranean coast of Israel during 2006 and 2007, and along a transect from Israel to Crete in September 2008. Analyses of time-series data revealed expression of nifH genes from diazotrophs in nifH clusters I and II, including cyanobacterial bloom-formers Trichodesmium and diatom-Richelia intracellularis associations. However, nifH gene abundance and rates of N(2) fixation were very low in all size fractions measured (> 0.7 µm). Volumetric (15) N uptake ranged from below detection (∼ 36% of > 300 samples) to a high of 0.3 nmol N l(-1) d(-1) and did not vary distinctly with depth or season. Areal N(2) fixation averaged ∼ 1 to 4 µmol N m(-2) d(-1) and contributed only ∼ 1% and 2% of new production and ∼ 0.25% and 0.5% of primary production for the mixed (winter) and stratified (spring-fall) periods respectively. N(2) fixation rates along the 2008 east-west transect were also extremely low (0-0.04 nmol N l(-1) d(-1), integrated average 2.6 µmol N m(-2) d(-1) ) with 37% of samples below detection and no discernable difference between stations. We demonstrate that diazotrophy and N(2) fixation contribute only a minor amount of new N to the P impoverished eastern Mediterranean Sea.

Suggett, David J. et. al.-- Contribution to AME Special 2 ‘Progress and perspectives in aquatic primary productivity’.-- 13 pages, 5 figures, Bioassay experiments were performed to identify how growth of key groups within the microbial community was simultaneously limited by nutrient (nitrogen and phosphorus) availability during spring in the Gulf of Aqaba’s oceanic waters. Measurements of chlorophyll a (chl a) concentration and fast repetition rate (FRR) fluorescence generally demonstrated that growth of obligate phototrophic phytoplankton was co-limited by N and P and growth of facultative aerobic anoxygenic photoheterotropic (AAP) bacteria was limited by N. Phytoplankton exhibited an increase in chl a biomass over 24 to 48 h upon relief of nutrient limitation. This response coincided with an increase in photosystem II (PSII) photochemical efficiency (Fv/Fm), but was preceded (within 24 h) by a decrease in effective absorption crosssection (σPSII) and electron turnover time (τ). A similar response for τ and bacterio-chl a was observed for the AAPs. Consistent with the up-regulation of PSII activity with FRR fluorescence were observations of newly synthesized PSII reaction centers via low temperature (77K) fluorescence spectroscopy for addition of N (and N + P). Flow cytometry revealed that the chl a and thus FRR fluorescence responses were partly driven by the picophytoplankton (, We thank the Batsheva de Rothschild Foundation, Bar-Ilan University, the Moshe Shilo Center for Marine Biogeochemistry, and the staff of the Interuniversity Institute for funding and logistic support. Attendance at the workshop was also supported by an international travel grant from the Texas Institute of Oceanography (A.Q.) and EUROCEANS (D.J.S., E.V.D. and T.L.)

Transparent exopolymer particles (TEP) are ubiquitous in marine and freshwaters, and have been subject to intensive study by oceanographers and limnologists over the past 15 years. These microscopic organic particles (visualized by Alcian Blue staining for acid polysaccharides) may be considered a planktonic form of exopolymeric substances (EPS). Two aspects relating to the potential involvement of TEP as important agents in organic fouling and biofilm formation on membranes in desalination and wastewater treatment plants were investigated: the efficiency of pretreatment processes in decreasing the amounts of TEP reaching the RO membranes at the Adom desalination plant, Ashkelon; and the active involvement of TEP in the early stages of biofilm formation. This study revealed that although pretreatment at the desalination plant lowered the levels of water quality parameters such as chlorophyll and SDI by ~90% relative to input, TEP concentrations were only decreased by ~30% upstream of the RO membranes. To ...