Photoinactivation of virus on iron-oxide coated sand: enhancing inactivation in sunlit waters

Adsorption onto iron oxides can enhance the removal of waterborne viruses in constructed wetlands and soils. If reversible adsorption is not coupled with inactivation, however, infective viruses may be released when changes in solution conditions cause desorption. The goals of this study were to investigate the release of infective bacteriophages MS2 and ΦX174 (two human viral indicators) after adsorption onto an iron oxide coated sand (IOCS), and to promote viral inactivation by exploiting the photoreactive properties of the IOCS. The iron oxide coating greatly enhanced viral adsorption (adsorption densities up to ∼10⁹ infective viruses/g IOCS) onto the sand, but had no affect on infectivity. Viruses that were adsorbed onto IOCS under control conditions (pH 7.5, 10 mM Tris, 1250 μS/cm) were released into solution in an infective state with increases in pH and humic acid concentrations. The exposure of IOCS-adsorbed MS2 to sunlight irradiation caused significant inactivation via a photocatalytic mechanism in both buffered solutions and in wastewater samples (4.9 log₁₀ and 3.3 log₁₀ inactivation after 24-h exposure, respectively). Unlike MS2, ΦX174 inactivation was not enhanced by photocatalysis. In summary, IOCS enhanced the separation of viruses from the water column, and additionally provided a photocatalytic mechanism to promote inactivation of one of the surrogates studied. These qualities make it an attractive option for improving viral control strategies in constructed wetlands.     

Virus inactivation by silver doped titanium dioxide nanoparticles for drinking water treatment

Photocatalytic inactivation of viruses and other microorganisms is a promising technology that has been increasingly utilized in recent years. In this study, photocatalytic silver doped titanium dioxide nanoparticles (nAg/TiO₂) were investigated for their capability of inactivating Bacteriophage MS2 in aqueous media. Nano-sized Ag deposits were formed on two commercial TiO₂ nanopowders using a photochemical reduction method. The MS2 inactivation kinetics of nAg/TiO₂ was compared to the base TiO₂ material and silver ions leached from the catalyst. The inactivation rate of MS2 was enhanced by more than 5 fold depending on the base TiO₂ material, and the inactivation efficiency increased with increasing silver content. The increased production of hydroxyl free radicals was found to be responsible for the enhanced viral inactivation.     

Pulsed ultraviolet light inactivation of Pseudomonas aeruginosa and Staphylococcus aureus biofilms

Microbial biofilms are complex communities that form when planktonic bacterial species attach to surfaces in many settings where they can provide a source of pathogenicity. The relative ineffectiveness of conventional disinfectants such as free chlorine and monochloramine for the inactivation of some species found in water has led to evaluation of alternative disinfectants for drinking and wastewater treatment. In recent years, novel pulsed power electrotechnologies have been introduced and are being considered as possible alternatives to current methods for inactivating problematic species in water. This study focuses on the ultraviolet (UV) inactivation of bacterial biofilms using a pulsed UV light approach as a potential disinfection method for water treatment operating systems. Biofilms were stimulated to form attached to polyvinyl chloride coupons using a recommended Centre for Disease Control biofilm reactor followed by exposure to a range of UV doses. Findings show that this method is highly effective at inactivating both planktonic and biofilm cells with significant inactivation rates obtained for both test species. Specifically, a 7.2 and 5.9 log₁₀ inactivation was achieved with up to 21.6 μJ/cm² UV for Pseudomonas aeruginosa and Staphylococcus aureus, respectively. Findings from this study highlight the effectiveness of pulsed UV for the inactivation of Pseudomonas biofilms among other test species. Research conducted by this group suggests that this pulsed UV system may offer a useful method for the disinfection of drinking and wastewater supplies.     

Source water quality effects on monochloramine inactivation of adenovirus, coxsackievirus, echovirus, and murine norovirus

There is a need for more information regarding monochloramine disinfection efficacy for viruses in water. In this study, monochloramine disinfection efficacy was investigated for coxsackievirus B5 (CVB5), echovirus 11 (E11), murine norovirus (MNV), and human adenovirus 2 (HAdV2) in one untreated ground water and two partially treated surface waters. Duplicate disinfection experiments were completed at pH 7 and 8 in source water at concentrations of 1 and 3 mg/L monochloramine at 5 and 15 °C. The Efficiency Factor Hom (EFH) model was used to calculate CT values (mg-min/L) required to achieve 2-, 3-, and 4-log₁₀ reductions in viral titers. In all water types, monochloramine disinfection was most effective for MNV, with 3-log₁₀ CT values at 5 °C ranging from 27 to 110. Monochloramine disinfection was least effective for HAdV2 and E11, depending on water type, with 3-log₁₀ CT values at 5 °C ranging from 1200 to 3300 and 810 to 2300, respectively. Overall, disinfection proceeded faster at 15 °C and pH 7 for all water types. Inactivation of the study viruses was significantly different between water types, but there was no indication that overall disinfection efficacy was enhanced or inhibited in any one water type. CT values for HAdV2 in two types of source water exceeded federal CT value recommendations in the US. The results of this study demonstrate that water quality impacts the inactivation of viruses and should be considered when developing chloramination plans.     

Inactivation of norovirus by chlorine disinfection of water

In an effort to validate previous research suggesting remarkable resistance of norovirus to free chlorine disinfection, we characterized the disinfection response of purified and dispersed Norwalk virus (NV) by bench-scale free chlorine disinfection using RT-PCR for virus assays. The inactivation of NV by two doses of free chlorine (1 and 5 mg/L) at pH 6 and 5 °C based on two RT-PCR assays was similar to that of coliphage MS2, but much faster than that of poliovirus 1. Despite the underestimation of virus inactivation by RT-PCR assays, the predicted CT values for NV based on RT-PCR assays are lower than the ones for most other important waterborne viruses and the CT guidelines for chlorine disinfection of viruses under the Surface Water Treatment Rule by the United States Environmental Protection Agency. Overall, the results of this study indicate that NV is not highly resistant to free chlorine disinfection as suggested by previous research and it is likely that NV contamination of drinking water can be controlled by adequate free chlorine disinfection practices with provision of proper pre-treatment processes before chlorination.     

Solar water disinfection (SODIS) of Escherichia coli, Enterococcus spp., and MS2 coliphage: effects of additives and alternative container materials

The use of alternative container materials and added oxidants accelerated the inactivation of MS2 coliphage and Escherichia coli and Enterococcus spp. bacteria during solar water disinfection (SODIS) trials. Specifically, bottles made from polypropylene copolymer (PPCO), a partially UVB-transparent plastic, resulted in three-log inactivation of these organisms in approximately half the time required for disinfection in bottles made from PET, polycarbonate, or Tritan^®, which absorb most UVB light. Furthermore, the addition of 125 mg/L sodium percarbonate in combination with either citric acid or copper plus ascorbate tended to accelerate inactivation by factors of 1.4-19. Finally, it was observed that the inactivation of E. coli and enterococci derived from local wastewater was far slower than the inactivation of laboratory-cultured E. coli and Enterococcus spp., while the inactivation of MS2 was slowest of all. These results highlight the importance of UVB in SODIS under certain conditions, and also the greater sunlight resistance of some viruses and of bacteria of fecal origin, as compared to the laboratory-cultured bacteria commonly used to model their inactivation. Furthermore, this study illustrates promising new avenues for accelerating the inactivation of bacteria and viruses by solar disinfection.     

UV inactivation and resistance of rotavirus evaluated by integrated cell culture and real-time RT-PCR assay

Rotaviruses are double-stranded RNA viruses which are among the most resistant waterborne enteric viruses to UV disinfection. An integrated cell culture and real-time RT-PCR (ICC real-time RT-PCR) assay was developed to detect the infectivity of rotaviruses in water, which uses real-time RT-PCR to detect RNA produced by infectious rotaviruses during replication in host cells. Detection of rotaviral RNA in host cells provides direct evidence of the presence of infectious rotavirus rather than just the presence of rotavirus RNA. Using this newly developed method, the inactivation and resistance of rotavirus to UV treatments at various doses was evaluated. With an initial concentration of 2 × 10⁴ PFU/ml simian rotavirus (SA11), a first-order linear relationship was obtained at UV dose range of 0-120 mJ cm⁻², and the inactivation rate constant was estimated to be 0.0343 cm² mJ⁻¹ (R² = 0.966). The dose-inactivation curve tailed off and reached plateau as the UV dose increased from 120 to 360 mJ cm⁻², indicating resistance phenomena of sub-populations of SA11 at very high UV doses. A maximal reduction of 4.8 log₁₀ was observed. Through parallel comparison with traditional culture assay, the ICC real-time RT-PCR method demonstrated more effective, sensitive and faster infectivity detection of rotavirus and, the results reveal that rotaviruses are more resistant to UV irradiation than previously reported with traditional cell culture assays.     

Inactivation and surface interactions of MS-2 bacteriophage in a TiO2 photoelectrocatalytic reactor

Inactivation of MS-2 bacteriophage in a TiO₂ photoelectrocatalytic system was evaluated, wherein TiO₂ particles were coated onto an indium tin oxide (ITO) electrode and an electrical potential was applied under black light blue (BLB) irradiation. MS-2 phage inactivation was greatly enhanced by anodic potential, whereas cathodic potential completely inhibited inactivation. Experiments performed with radical scavengers showed that inactivation was primarily caused by hydroxyl radicals, both in the bulk phase and on the TiO₂ surface. Application of positive potential to the electrode was found to result in two distinct beneficial effects: (i) electrostatic attraction between the negatively charged viral capsid and catalyst surface, causing improved usage of surface-bound hydroxyl radical, in comparison to conventional TiO₂ photocatalytic disinfection; and (ii) higher reactive oxygen species production. Results also suggest that inactivation of various microorganisms including Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Bacillus subtilis spores and Cryptosporidium parvum oocyst was enhanced via positive potential induction to TiO₂.     

Inactivation of bacteriophage MS2 upon exposure to very low concentrations of chlorine dioxide

This study investigates the effects of very low concentrations of ClO₂ applied in drinking water practice on the inactivation of bacteriophage MS2. Concentrations of 0.5 mg/L, 0.1 mg/L and 0.02 mg/L ClO₂ inactivated at least 5 log units of MS2 after an exposure time of approximately 20, 50 and 300 min respectively. When the ClO₂ concentration was as low as 0.005 mg/L, inactivation of 1 log unit MS2 was observed after 300 min exposure. Increasing the contact time to 24 h did not increase the inactivation any further. Non-linear inactivation kinetics (tailing) were observed for all conditions tested. Repeated addition of MS2 to the reactor showed that tailing was not caused by a reduction of the biocidal effect of ClO₂ during disinfection. The Modified Chick-Watson, the Efficiency Factor Hom (EFH) model and the Modified Cerf model, a modification of the two-fraction Cerf model, were fitted to the non-linear inactivation curves. Both the EFH and the modified Cerf model did fit accurately to the inactivation data of all experiments. The good fit of the Modified Cerf model supports the hypothesis of the presence of two subpopulations. Our study showed that ClO₂ is an effective disinfectant against model organism MS2, also at the low concentrations applied in water treatment practice. The inactivation kinetics followed a biphasic pattern due to the presence of a more ClO₂-resistant subpopulation of MS2 phages, either caused by population heterogeneity or aggregation/adhesion of MS2.     

Disinfection of Legionella pneumophila by ultrasonic treatment with TiO2

An ultrasonic treatment system, using a TiO₂ photocatalyst, was used to disinfect Legionella pneumophila. A kinetic study of the process indicates that TiO₂ significantly improves the disinfection process. The concentrations of viable cells were reduced to 6% of the initial concentrations in the presence of 0.2 g/ml TiO₂ after a 30 min of treatment period, while only an 18% reduction was observed in the absence of TiO₂. The potency of the disinfection could be enhanced, to some extent, by increasing the amount of TiO₂ used. Cell concentrations were decreased by an order of 3 within 30 min of treatment in the presence of 1.0 g/ml TiO₂. The disinfection power in the presence of TiO₂ versus Al₂O₃ was also compared and the findings showed that TiO₂ induced a higher cell killing. No significant effect of initial cell concentration on the disinfection was found in the range of 10²-10⁷ CFU/ml after a 30 min of treatment period. The mechanism of cell killing was investigated by examining the effects of OH radical scavengers such as ascorbic acid, histidine and glutathione. The disinfection power was reduced in samples that contained these radical scavengers, thus indicating the importance of OH radicals.     

Combined permeable pavement and photocatalytic titanium dioxide oxidation system for urban run‐off treatment and disinfection

Permeable pavement systems (PPS) are frequently associated with high removal efficiencies for water quality parameters. Their effluent can, therefore, be recycled, for example, for sprinkling onto gardens. Nevertheless, some stakeholders fear that potentially pathogenic organisms within the treated run‐off could be too high, and therefore they request disinfection before recycling. The aim of this paper is, therefore, to assess the efficiency of a batch flow combined titanium dioxide (TiO₂) and ultraviolet (UV) light photocatalytic reactor in removing water‐borne microbial contaminants from the effluent of PPS. Combined TiO₂ and UV photocatalytic reaction times between 80 and 100 min were required for the complete removal of Escherichia coli, total coliforms and faecal Streptococci, which had mean initial counts of 1.5 × 10⁷, 4.4 × 10⁶ and 6.9 × 10⁵ colony‐forming units (CFU) per 100 mL, respectively. In comparison, UV disinfection alone resulted in insignificant microbial removal. Suspended TiO₂ powder was more effective than small immobilised TiO₂ crystals.     

Photocatalytic oxidation of DBP precursors using UV with suspended and fixed TiO2

French River water (Nova Scotia, Canada) was separated into six different natural organic matter (NOM) fractions, including hydrophobic acids, bases and neutrals and hydrophilic acids, bases and neutrals. The raw water, as well as each of the NOM fractions were analysed for disinfection by-product (DBP) formation potential before and after advanced oxidation with UV/TiO₂ to determine the efficacy of this treatment for the removal of DBP precursors. The UV/TiO₂ treatment was carried out with a nanostructured thin film (NSTF), coated with TiO₂ which is compared with the use of a TiO₂ suspension. For the raw river water, removals of total trihalomethane formation potential (TTHMFP) and total haloacetic acid formation potential (THAA₉FP) were found to be approximately 20% and 90%, respectively, with 50 mJ/cm² UV exposure and 1 mg/L TiO₂. For the fractionated samples, approximately 75% of both trihalomethane (THM) and haloacetic acid (HAA) precursors were found to be associated with the hydrophobic acid fraction. For this individual fraction the same UV/TiO₂ treatments exhibited approximately 20-25% removal of both TTHMFP and THAA₉FP, suggesting that the fractionation process may have affected the treatability of HAA precursors or may have altered the results of the oxidation processes.     

Assessment of the removal and inactivation of influenza viruses H5N1 and H1N1 by drinking water treatment

Since 2003, there has been significant concern about the possibility of an outbreak of avian influenza virus subtype H5N1. Moreover, in the last few months, a pandemic of a novel swine-origin influenza A virus, namely A(H1N1), has already caused hundreds of thousands of human cases of illness and thousands of deaths. As those viruses could possibly contaminate water resources through wild birds excreta or through sewage, the aim of our work was to find out whether the treatment processes in use in the drinking water industry are suitable for eradicating them. The effectiveness of physical treatments (coagulation-flocculation-settling, membrane ultrafiltration and ultraviolet) was assessed on H5N1, and that of disinfectants (monochloramine, chlorine dioxide, chlorine, and ozone) was established for both the H5N1 and H1N1 viruses.Natural water samples were spiked with human H5N1/H1N1 viruses. For the coagulation-settling experiments, raw surface water was treated in jar-test pilots with 3 different coagulating agents (aluminum sulfate, ferric chloride, aluminum polychorosulfate). Membrane performance was quantified using a hollow-fiber ultrafiltration system. Ultraviolet irradiation experiments were conducted with a collimated beam that made it possible to assess the effectiveness of various UV doses (25-60 mJ/cm²). In the case of ozone, 0.5 mg/L and 1 mg/L residual concentrations were tested with a contact time of 10 min. Finally, for chlorine, chlorine dioxide and monochloramine treatments, several residual oxidant target levels were tested (from 0.3 to 3 mg/L) with contact times of 5-120 min. The infectivity of the H5N1 and H1N1 viruses in water samples was quantified in cell culture using a microtiter endpoint titration.The impact of coagulation-settling on the H5N1 subtype was quite low and variable. In contrast, ultrafiltration achieved more than a 3-log reduction (and more than a 4-log removal in most cases), and UV treatment was readily effective on its inactivation (more than a 5-log inactivation with a UV dose of 25 mJ/cm²). Of the chemical disinfection treatments, ozone, chlorine and chlorine dioxide were all very effective in inactivating H5N1 and H1N1, whereas monochloramine treatment required higher doses and longer contact times to achieve significant reductions.Our findings suggest that the water treatment strategies that are currently used for surface water treatment are entirely suitable for removing and/or inactivating influenza A viruses. Appropriate preventive actions can be defined for single disinfection treatment plants.     

Influence of humic substances on the ultraviolet disinfection of surface waters

The efficiency of ultraviolet (UV) disinfection and the regrowth potential of total coliforms were investigated in humic waters. Experiments were conducted according to an experimental design in which a UV radiation dose range of 68–681 mW s/cm² was applied to waters containing various concentrations (0–10 mg/L) of fulvic acid. Experimental results strongly suggested that the harmful effect of UV radiation on bacteria was diminished in humic waters due to absorption of UV light. Increasing concentrations of fulvic acid appeared to enhance its influence with elevated doses of UV radiation. Measured inactivation responses were on the order of 1–6 log₁₀ units for the UV dose range used. Comparison of k (inactivation coefficient) values for nonhumic water to highly humic water indicated that k decreased proportionally as the fulvic acid concentration of water was increased. The results of dark‐incubation tests indicated significant regrowth of bacteria in fulvic acid‐containing waters.     

Concurrent filtration and solar photocatalytic disinfection/degradation using high-performance Ag/TiO2 nanofiber membrane

A facile polyol synthesis was used for the deposition of Ag nanoparticles on electrospun TiO₂ nanofibers for the subsequent fabrication of Ag/TiO₂ nanofiber membrane. The permeate flux of the Ag/TiO₂ nanofiber membrane was remarkably high compared to commercial P25 deposited membrane. The Ag/TiO₂ nanofiber membrane achieved 99.9% bacteria inactivation and 80.0% dye degradation under solar irradiation within 30 min. The Ag/TiO₂ nanofiber membrane also showed excellent antibacterial capability without solar irradiation. Considering the excellent intrinsic antibacterial activity and high-performance photocatalytic disinfection/degradation under solar irradiation, this novel membrane proved to have promising applications in water purification industry.     

Low- and medium-pressure UV inactivation of microsporidia Encephalitozoon intestinalis

Newly recognized waterborne pathogens such as microsporidia are being detected in the world's water supplies with increasing frequency. Many of these organisms have been shown to cause negative health impacts for both immunocompetent as well as immunocompromised individuals. It is imperative that these emerging pathogens be investigated for their ability to resist both traditional and novel disinfection technologies that are currently in use or under consideration for drinking water treatment. Low- and medium pressure UV light is at the cutting edge of disinfection technologies for the drinking water industry. While previous UV disinfection studies have focused on the inactivation of Cryptosporidium and Giardia as well as viruses and common bacteria, this research reports the ability of low- and medium pressure UV light to inactivate > 3.6 log10 of microsporidia Encephalitozoon intestinalis spores at a dose of 6 mJ/cm2 or higher as determined using a cell culture approach.     

Inactivation of environmental mycobacteria by free chlorine and UV

The resistance of environmental mycobacteria (EM) against chlorine and ultraviolet (UV) was evaluated for determination of the Ct value and UV dose to inactivate EM. Chlorine disinfection experiments were done on Mycobacterium fortuitum in oxidant demand-free buffered water at the worst condition (pH 8.5, 4 °C) and normal condition (pH 7.0, 20 °C). The Ct value for 3 log inactivation of M. fortuitum was 600 times greater than that of Escherichia coli. UV experiments were performed for various species of Mycobacterium avium, M. fortuitum, Mycobacterium intracellulare, and Mycobacterium lentiflavum. A UV collimated beam device was used for irradiation of four species suspended in phosphate buffered saline with doses of 5, 10, 20, 50, and 100 mJ/cm². UV sensitivity of mycobacteria was species-specific. More than 3 log of M. avium, M. intracellulare, and M. lentiflavum could be inactivated at 20 mJ/cm², whereas M. fortuitum was so resistant that 3 log inactivation required a dose of more than 50 mJ/cm². Mycobacteria were found 2-10 times more resistant to UV than E. coli for 3 log inactivation. There was no significant difference in the inactivation of mycobacteria with either low-pressure or medium-pressure UV irradiation.     

Effect of particles and bioflocculation on ultraviolet disinfection of Escherichia coli

Presence of particles is known to decrease the effectiveness of ultraviolet (UV) disinfection by shielding the targeted microorganisms from UV light. This study aims to provide an in-depth understanding on the effect of particles and flocs on UV disinfection by using a stable, well-defined and well-controlled synthetic system that can simulate the bioflocculation of particles and microorganisms in water and wastewater samples. The synthetic system was created by using Escherichia coli, latex particles (1, 3.2, 11, 25, and 45 μm), alginate, and divalent cations; and the bioflocculation of particles was achieved naturally, as it would occur in the environment, without using chemical coagulants. E. coli was quantified before and after UV disinfection using membrane filtration. Even in the absence of particles, some of the self-aggregated E. coli could survive a UV dose of 90 mJ/cm². E. coli inactivation levels measured in the presence of particles were lower than the inactivation levels measured in the absence of particles. At low UV doses (<9 mJ/cm²), neither particle size nor degree of flocculation had a significant effect on the inactivation of E. coli. Particle size had a significant effect on the inactivation of E. coli only at high UV doses (80 mJ/cm²), and larger particles (e.g., 25 μm) protected bacteria more compared to smaller particles (e.g., 3.2 and 11 μm). What size of particles flocs were made of (3.2, 11, and 25 μm) did not make a significant difference on the inactivation levels of E. coli. For 3.2 μm particles, there was no significant difference in E. coli inactivation between non-flocculated and flocculated samples at any UV dose. For 11 and 25 μm particles, there was a significant difference in E. coli inactivation between non-flocculated and flocculated samples at 80 mJ/cm². Degree of flocculation became a significant factor in determining the number of surviving bacteria only at high UV doses and only for larger particles.     

Bactericidal efficiency and mode of action: a comparative study of photochemistry and photocatalysis

In order to compare the disinfection potential of photocatalysis and photochemistry, the effects of these two processes on bacteria in water were investigated under exposure to UV-A and UV-C. The well-known bacterial model Escherichia coli (E. coli) was used as the experimental organism. Radiation exposure was produced with an HPK 125 W lamp and the standard TiO₂ Degussa P-25 was used as the photocatalyst.Firstly, the impact of photocatalysis and photochemistry on the cultivability of bacterial cells was investigated. UV-A radiation resulted in low deleterious effects on bacterial cultivability but generated colonies of size smaller than average. UV-C photocatalysis demonstrated a greater efficiency than UV-A photocatalysis in altering bacterial cultivability. From a cultivability point of view only, UV-C radiation appeared to be the most deleterious treatment.A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit was then used to assess the modifications in bacterial membrane permeability. UV-A radiation did not induce any alterations in bacterial permeability for 420 min of exposure whereas only a few minutes of exposure to UV-C radiation, with the same total radiance intensity, induced total loss of permeability. Moreover, after 20 and 60 min of exposure to UV-C and UV-A photocatalysis respectively, all bacteria lost their membrane integrity, suggesting that the bacterial envelope is the primary target of reactive oxygen species (ROS) generated at the surface of TiO₂ photocatalyst. These results were further confirmed by the formation of malondialdehyde (MDA) during the photocatalytic inactivation of bacterial cells and suggest that destruction of the cell envelope is a key step in the bactericidal action of photocatalysis. The oxidation of bacterial membrane lipids was also correlated with the monitoring of carboxylic acids, which can be considered as representatives of lipid peroxidation by-products.Finally, damages to bacterial morphology induced by UV-C photocatalysis and photochemistry were investigated through Scanning electron microscopy (SEM). Bacterial cells were observed on microscopy pictures at exposure durations corresponding to a loss of cultivability. After 90 min of exposure to UV-C radiation, bacterial cells showed little alteration of their outer membrane whereas they suffered deep deleterious damages under UV-C photocatalysis exposure.