Impacts of Desalination Brine Discharge on Benthic Ecosystems.

Seawater reverse osmosis (SWRO) desalination facilities produce freshwater and, at the same time, discharge hypersaline brine that often includes various chemical additives such as antiscalants and coagulants. This dense brine can sink to the sea bottom and creep over the seabed, reaching up to 5 km from the discharge point. Previous reviews have discussed the effects of SWRO desalination brine on various marine ecosystems, yet little attention has been paid to the impacts on benthic habitats. This review comprehensibly discusses the effects of SWRO brine discharge on marine benthic fauna and flora. We review previous studies that indicated a suite of impacts by SWRO brine on benthic organisms, including bacteria, seagrasses, polychaetes, and corals. The effects within the discharge mixing zones range from impaired activities and morphological deformations to changes in the community composition. Recent modeling work demonstrated that brine could spread over the seabed, beyond the mixing zone, for up to several tens of kilometers and impair nutrient fluxes from the sediment to the water column. We also provide a possible perspective on brine's impact on the biogeochemical process within the mixing zone subsurface. Desalination brine can infiltrate into the sandy bottom around the discharge area due to gravity currents. Accumulation of brine and associated chemical additives, such as polyphosphonate-based antiscalants and ferric-based coagulants in the porewater, may change the redox zones and, hence, impact biogeochemical processes in sediments. With the demand for drinking water escalating worldwide, the volumes of brine discharge are predicted to triple during the current century. Future efforts should focus on the development and operation of viable technologies to minimize the volumes of brine discharged into marine environments, along with a change to environmentally friendly additives. However, the application of these technologies should be partly subsidized by governmental stakeholders to safeguard coastal ecosystems around desalination facilities.

Microporous Polyethersulfone Membranes Grafted with Zwitterionic Polymer Brushes Showing Microfiltration Permeance and Ultrafiltration Bacteriophage Removal.

Virus removal from water using microfiltration (MF) membranes is of great interest but remains challenging owing to the membranes' mean pore sizes typically being significantly larger than most viruses. We present microporous membranes grafted with polyzwitterionic brushes (N-dimethylammonium betaine) that combine bacteriophage removal in the range of ultrafiltration (UF) membranes with the permeance of MF membranes. Brush structures were grafted in two steps: free-radical polymerization followed by atom transfer radical polymerization (ATRP). Attenuated total reflection Fourier transform infrared (ATR-FTIR) and X-ray photoelectron (XPS) verified that grafting occurred at both sides of the membranes and that the grafting increased with increasing the zwitterion monomer concentration. The log reduction values (LRVs) of the pristine membrane increased from less than 0.5 LRV for T4 (∼100 nm) and NT1 (∼50 nm) bacteriophages to up to 4.5 LRV for the T4 and 3.1 LRV for the NT1 for the brush-grafted membranes with a permeance of about 1000 LMH/bar. The high permeance was attributed to a high-water fraction in the ultra-hydrophilic brush structure. The high measured LRVs of the brush-grafted membranes were attributed to enhanced bacteriophages exclusion from the membrane surface and entrapment of the ones that penetrated the pores due to the membranes' smaller mean pore-size and cross-section porosity than those of the pristine membrane, as seen by scanning electron microscopy (SEM) and measured using liquid-liquid porometry. Micro X-ray fluorescence (µ-XRF) spectrometry and nanoscale secondary ion mass spectrometry showed that 100 nm Si-coated gold nanospheres accumulated on the surface of the pristine membrane but not on the brush-coated membrane and that the nanospheres that penetrated the membranes were entrapped in the brush-grafted membrane but passed the pristine one. These results corroborate the LRVs obtained during filtration experiments and support the inference that the increased removal was due to a combined exclusion mechanism and entrapment. Overall, these microporous brush-grafted membranes show potential for use in advanced water treatment.

Antiscalants used in the desalination industry impact the physiology of the coral Montipora capricornis.

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.

Discharge of Polyphosphonate-Based Antiscalants via Desalination Brine: Impact on Seabed Nutrient Flux and Microbial Activity.

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 ex-situ core benthocosms, to which we added brine from the dispersion area in excess salinities of ∼3% and 5% above natural levels. A higher influx of dissolved organic phosphorus (∼6-fold) and an efflux of dissolved organic carbon (∼1.7-fold) were measured in the brine-amended cores relative to the controls. This was accompanied by increased oxygen consumption (15%) and increased microbial activity (∼1.5-6.5-fold). Field observations support the results from experimental manipulations, yielding ∼4.5-fold higher microbial activity rates around the brine plume compared to uninfluenced locations. Our results imply that desalination brine can alter sedimentary processes affecting benthic nutrients inventories. Moreover, we show that brine acts as a vector of anthropogenic P, stimulating microbial activity in the sediment-water interface.

Biofilm Formation and Biofouling Development on Different Ultrafiltration Membranes by Natural Anaerobes from an Anaerobic Membrane Bioreactor.

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.

Planktonic Aggregates as Hotspots for Heterotrophic Diazotrophy: The Plot Thickens.

Biological dinitrogen (N2) fixation is performed solely by specialized bacteria and archaea termed diazotrophs, introducing new reactive nitrogen into aquatic environments. Conventionally, phototrophic cyanobacteria are considered the major diazotrophs in aquatic environments. However, accumulating evidence indicates that diverse non-cyanobacterial diazotrophs (NCDs) inhabit a wide range of aquatic ecosystems, including temperate and polar latitudes, coastal environments and the deep ocean. NCDs are thus suspected to impact global nitrogen cycling decisively, yet their ecological and quantitative importance remain unknown. Here we review recent molecular and biogeochemical evidence demonstrating that pelagic NCDs inhabit and thrive especially on aggregates in diverse aquatic ecosystems. Aggregates are characterized by reduced-oxygen microzones, high C:N ratio (above Redfield) and high availability of labile carbon as compared to the ambient water. We argue that planktonic aggregates are important loci for energetically-expensive N2 fixation by NCDs and propose a conceptual framework for aggregate-associated N2 fixation. Future studies on aggregate-associated diazotrophy, using novel methodological approaches, are encouraged to address the ecological relevance of NCDs for nitrogen cycling in aquatic environments.

Contribution of Heterotrophic Diazotrophs to N2 Fixation in a Eutrophic River: Free-Living vs. Aggregate-Associated.

Recent studies have indicated that heterotrophic diazotrophs are highly diverse and fix N2 in aquatic environments with potentially adverse conditions for diazotrophy, such as oxic and rich in total nitrogen. In this study, we compared the activity and diversity of heterotrophic diazotrophs associated with aggregates (>12 µm) to free-living cells in the eutrophic Qishon River during the winter and summer seasons. Overall, measured heterotrophic N2 fixation rates in the Qishon River ranged between 2.6-3.5 nmol N L-1 d-1. Heterotrophic N2 fixation was mainly associated with aggregates in the summer samples (74 ± 24%), whereas during the winter the bulk diazotrophic activity was mostly ascribed to the free-living fraction (90 ± 6%). In addition, immunolabeled micrographs indicated the presence of aggregate-associated heterotrophic diazotrophs in both seasons, while phototrophic diazotrophs were also captured during the winter. The richness of free-living and aggregate-associated heterotrophic diazotrophs were overall similar, yet the evenness of the later was significantly smaller, suggesting that few of the species gained advantage from particle lifestyle. The differences in the activity, micro-localization and diversity of the diazotrophic community were mostly attributed to spatiotemporal changes in the ambient C:N ratios (total organic carbon, TOC: total nitrogen) and the TOC concentrations. Taken together, our results shed new light on the contribution of heterotrophic diazotroph associated with aggregates to total heterotrophic N2 fixation in oxic, highly eutrophic aquatic environments.

Jellyfish swarm impair the pretreatment efficiency and membrane performance of seawater reverse osmosis desalination.

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.

Impacts of sewage outbursts on seawater reverse osmosis desalination.

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.

Bursting out: linking changes in nanotopography and biomechanical properties of biofilm-forming Escherichia coli to the T4 lytic cycle.

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.

Tracking SARS-CoV-2 RNA through the Wastewater Treatment Process.

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.

Heterotrophic Nitrogen Fixation at the Hyper-Eutrophic Qishon River and Estuary System.

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.

Impact of Hydrodynamics on the First Stages of Biofilm Formation in Forward Osmosis with Spacers.

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 (<100 cells mm-2). Imaging at 30 s intervals revealed three phases (i.e., lag, exponential, and linear) in the development of deposition over time. Quantification of spatial deposition patterns showed that an increase in the ratio of permeate water flux to crossflow velocity led to a homogeneous deposition, while a decrease had the opposite effect. The insights of this research indicate that an appropriate choice of hydrodynamic conditions can minimize bacteria accumulation prior to biofilm formation in new and cleaned FO membrane systems treating water of high fouling propensity.

Phytoplankton response to N-rich well amelioration brines: A mesocosm study from the southeastern Mediterranean Sea.

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.

Direct Detection of Heterotrophic Diazotrophs Associated with Planktonic Aggregates.

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.

Comparing membrane and spacer biofouling by Gram-negative Pseudomonas aeruginosa and Gram-positive Anoxybacillus sp. in forward osmosis.

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.

Chronic effects of brine discharge form large-scale seawater reverse osmosis desalination facilities on benthic bacteria.

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 (<1.4 Km2) around the discharge point. Namely, that the effects on benthic bacteria are prominent at the brine mixing zone and change according to the discharge method used to disperse the brine as well as local stressors (e.g., eutrophication and elevated water temperature). Our results contribute new insights on the effects of desalination-brine to benthic microbes, while providing scientifically-based aspects on the ecological impacts of brine dispersion for decision makers.

Development of anti-biofouling feed spacers to improve performance of reverse osmosis modules.

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.

Membrane Distillation Biofouling: Impact of Feedwater Temperature on Biofilm Characteristics and Membrane Performance.

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.

Physisorption and chemisorption of T4 bacteriophages on amino functionalized silica particles.

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.