Mitigation of viruses of concern and bacteriophage surrogates via common unit processes for water reuse: A meta-analysis.

Water reuse is a growing global reality. In regulating water reuse, viruses have come to the fore as key pathogens due to high shedding rates, low infectious doses, and resilience to traditional wastewater treatments. To demonstrate the high log reductions required by emerging water reuse regulations, cost and practicality necessitate surrogates for viruses for use as challenge organisms in unit process evaluation and monitoring. Bacteriophage surrogates that are mitigated to the same or lesser extent than viruses of concern are routinely used for individual unit process testing. However, the behavior of these surrogates over a multi-barrier treatment train typical of water reuse has not been well-established. Toward this aim, we performed a meta-analysis of log reductions of common bacteriophage surrogates for five treatment processes typical of water reuse treatment trains: advanced oxidation processes, chlorination, membrane filtration, ozonation, and ultraviolet (UV) disinfection. Robust linear regression was applied to identify a range of doses consistent with a given log reduction of bacteriophages and viruses of concern for each treatment process. The results were used to determine relative conservatism of surrogates. We found that no one bacteriophage was a representative or conservative surrogate for viruses of concern across all multi-barrier treatments (encompassing multiple mechanisms of virus mitigation). Rather, a suite of bacteriophage surrogates provides both a representative range of inactivation and information about the effectiveness of individual processes within a treatment train. Based on the abundance of available data and diversity of virus treatability using these five key water reuse treatment processes, bacteriophages MS2, phiX174, and Qbeta were recommended as a core suite of surrogates for virus challenge testing.

A Comparison of Porphyrin Photosensitizers in Photodynamic Inactivation of RNA and DNA Bacteriophages.

Effective broad-spectrum antiviral treatments are in dire need as disinfectants and therapeutic alternatives. One such method of disinfection is photodynamic inactivation, which involves the production of reactive oxygen species from dissolved oxygen in response to light-stimulated photosensitizers. This study evaluated the efficacy of functionalized porphyrin compounds for photodynamic inactivation of bacteriophages as human virus surrogates. A blue-light light emitting diode (LED) lamp was used to activate porphyrin compounds in aqueous solution (phosphate buffer). The DNA bacteriophages ΦX174 and P22 were more resistant to porphyrin TMPyP photodynamic inactivation than RNA bacteriophage fr, with increasing rates of inactivation in the order: ΦX174 << P22 << fr. Bacteriophage ΦX174 was therefore considered a resistant virus suitable for the evaluation of three additional porphyrins. These porphyrins were synthesized from TMPyP by inclusion of a central palladium ion (PdT4) and/or the addition of a hydrophobic C14 chain (PdC14 or C14). While the inactivation rate of bacteriophage ΦX174 via TMPyP was similar to previous reports of resistant viruses, ΦX174 inactivation increased by a factor of approximately 2.5 using the metalloporphyrins PdT4 and PdC14. The order of porphyrin effectiveness was TMPyP < C14 < PdT4 < PdC14, indicating that both Pd2+ ligation and C14 functionalization aided virus inactivation.

Electrocoagulation-electrooxidation for mitigating trace organic compounds in model drinking water sources.

In-situ water treatment can be accomplished using electrochemical treatments such as electrocoagulation (EC), which generates coagulants, and electrooxidation (EO), which generates oxidants (e.g., free chlorine and reactive oxygen species) via electrolysis using boron-doped diamond electrodes. In sequential EC-EO, EC can remove oxidant scavengers present in dissolved organic carbon (DOC), thereby improving the efficacy of downstream oxidation via EO. This study evaluated sequential EC-EO (and each process independently for comparison) for mitigating the trace organic compounds (TOrCs) acyclovir (ACY), trimethoprim (TMP), and benzyldimethyldecylammonium chloride (BAC-C10) in model groundwaters and surface waters. EO-only removed greater than 70% of ACY and TMP but negligible BAC-C10 in model groundwaters. In model surface waters, EO-only removed ∼55-75% BAC-C10, but had less removal for ACY and TMP (∼20-55%), primarily due to DOC interference. Sequential-EC-EO was investigated to better gauge the potential process improvement due to the addition of EC ahead of EO. EC removed 74 ± 7% DOC from model surface water and improved downstream EO treatment relative to EO-only by a factor of 3.4 for ACY, 1.7 for TMP, and 1.4 for BAC-C10. When treating model groundwater, EC-EO resulted in no improvement compared to EO-only for ACY and TMP. BAC-C10 removal was attributed to the particle separation step between EC and EO rather than electrochemical inputs. EO-only treatment was more energy efficient for model groundwater compared to model surface waters based on electrical energy per order (EEO) values. Sequential EC-EO further improved the energy efficiency for treating model river water.

Virus Isoelectric Point Estimation: Theories and Methods.

Much of virus fate, both in the environment and in physical/chemical treatment, is dependent on electrostatic interactions. Developing an accurate means of predicting virion isoelectric point (pI) would help to understand and anticipate virus fate and transport, especially for viruses that are not readily propagated in the lab. One simple approach to predicting pI estimates the pH at which the sum of charges from ionizable amino acids in capsid proteins approaches zero. However, predicted pIs based on capsid charges frequently deviate by several pH units from empirically measured pIs. Recently, the discrepancy between empirical and predicted pI was attributed to the electrostatic neutralization of predictable polynucleotide-binding regions (PBRs) of the capsid interior. In this paper, we review models presupposing (i) the influence of the viral polynucleotide on surface charge or (ii) the contribution of only exterior residues to surface charge. We then compare these models to the approach of excluding only PBRs and hypothesize a conceptual electrostatic model that aligns with this approach. The PBR exclusion method outperformed methods based on three-dimensional (3D) structure and accounted for major discrepancies in predicted pIs without adversely affecting pI prediction for a diverse range of viruses. In addition, the PBR exclusion method was determined to be the best available method for predicting virus pI, since (i) PBRs are predicted independently of the impact on pI, (ii) PBR prediction relies on proteome sequences rather than detailed structural models, and (iii) PBR exclusion was successfully demonstrated on a diverse set of viruses. These models apply to nonenveloped viruses only. A similar model for enveloped viruses is complicated by a lack of data on enveloped virus pI, as well as uncertainties regarding the influence of the phospholipid envelope on charge and ion gradients.

Improved Virus Isoelectric Point Estimation by Exclusion of Known and Predicted Genome-Binding Regions.

Knowledge of the isoelectric points (pIs) of viruses is beneficial for predicting virus behavior in environmental transport and physical/chemical treatment applications. However, the empirically measured pIs of many viruses have thus far defied simple explanation, let alone prediction, based on the ionizable amino acid composition of the virus capsid. Here, we suggest an approach for predicting the pI of nonenveloped viruses by excluding capsid regions that stabilize the virus polynucleotide via electrostatic interactions. This method was applied first to viruses with known polynucleotide-binding regions (PBRs) and/or three-dimensional (3D) structures. Then, PBRs were predicted in a group of 32 unique viral capsid proteome sequences via conserved structures and sequence motifs. Removing predicted PBRs resulted in a significantly better fit to empirical pI values. After modification, mean differences between theoretical and empirical pI values were reduced from 2.1 ± 2.4 to 0.1 ± 1.7 pH units.IMPORTANCE This model fits predicted pIs to empirical values for a diverse set of viruses. The results suggest that many previously reported discrepancies between theoretical and empirical virus pIs can be explained by coulombic neutralization of PBRs of the inner capsid. Given the diversity of virus capsid structures, this nonarbitrary, heuristic approach to predicting virus pI offers an effective alternative to a simplistic, one-size-fits-all charge model of the virion. The accurate, structure-based prediction of PBRs of the virus capsid employed here may also be of general interest to structural virologists.

Mechanisms of virus mitigation and suitability of bacteriophages as surrogates in drinking water treatment by iron electrocoagulation.

Emerging water treatment technologies using ferrous and zero-valent iron show promising virus mitigation by both inactivation and adsorption. In this study, iron electrocoagulation was investigated for virus mitigation in drinking water via bench-scale batch experiments. Relative contributions of physical removal and inactivation, as determined by recovery via pH 9.5 beef broth elution, were investigated for three mammalian viruses (adenovirus, echovirus, and feline calicivirus) and four bacteriophage surrogates (fr, MS2, P22, and ΦX174). Though no one bacteriophage exactly represented mitigation of the mammalian viruses in all water matrices, bacteriophage ΦX174 was the only surrogate that showed overall removal comparable to that of the mammalian viruses. Bacteriophages fr, MS2, and P22 were all more susceptible to inactivation than the three mammalian viruses, raising concerns about the suitability of these common surrogates as indicators of virus mitigation. To determine why some bacteriophages were particularly susceptible to inactivation, mechanisms of bacteriophage mitigation due to electrocoagulation were investigated. Physical removal was primarily due to inclusion in flocs, while inactivation was primarily due to ferrous iron oxidation. Greater electrostatic attraction, virus aggregation, and capsid durability were proposed as reasons for virus susceptibility to ferrous-based inactivation. Results suggest that overall treatment claims based on bacteriophage mitigation for any iron-based technology should be critically considered due to higher susceptibility of bacteriophages to inactivation via ferrous oxidation.

Sequential electrocoagulation-electrooxidation for virus mitigation in drinking water.

Electrochemical water treatment is a promising alternative for small-scale and remote water systems that lack operational capacity or convenient access to reagents for chemical coagulation and disinfection. In this study, the mitigation of viruses was investigated using electrocoagulation as a pretreatment prior to electrooxidation treatment using boron-doped diamond electrodes. This research is the first to investigate a sequential electrocoagulation-electrooxidation treatment system for virus removal. Bench-scale, batch reactors were used to evaluate mitigation of viruses in variable water quality via: a) electrooxidation, and b) a sequential electrocoagulation-electrooxidation treatment train. Electrooxidation of two bacteriophages, MS2 and ΦX174, was inhibited by natural organic matter and turbidity, indicating the probable need for pretreatment. However, the electrocoagulation-electrooxidation treatment train was beneficial only in the model surface waters employed. In model groundwaters, electrocoagulation alone was as good or better than the combined electrocoagulation-electrooxidation treatment train. Reduction of human echovirus was significantly lower than one or both bacteriophages in all model waters, though bacteriophage ΦX174 was a more representative surrogate than MS2 in the presence of natural organic matter and turbidity. Compared to conventional treatment by ferric salt coagulant and free chlorine disinfection, the electrocoagulation-electrooxidation system was less effective in model surface waters but more effective in model groundwaters. Sequential electrocoagulation-electrooxidation was beneficial for some applications, though practical considerations may currently outweigh the benefits.

Analysis of operational parameters, reactor kinetics, and floc characterization for the removal of estrogens via electrocoagulation.

Estrogenic compounds can cause human and ecological health issues and have been detected in surface and drinking water. In this research a reactor analysis determined the impact of operational parameters, the best fit kinetic model for the removal of estrone (E1), 17ß-estradiol (E2), estriol (E3), and 17α-ethynylestradiol (EE2) using a bench-top iron electrocoagulation reactor, and characterized the floc generated in-situ. The parameters investigated were current density, conductivity, stir rate, and polarity reversal. Estrogen removal correlated well with an increase in current density, while conductivity did not impact removal but did reduce potentials. High stir rates and frequent polarity reversal demonstrated greater removal. The operating parameters that achieved the greatest estrogen removal were a current density of 16.7 mA cm-2, conductivity of 1000 µS cm-1, stir rate of 500 rpm, and a polarity reversal time of 30 s. These parameters led to average removal efficiencies of 81%, 87%, 85%, and 97% for E1, E2, E3, and EE2, respectively. The removal data for all estrogenic compounds best fit a pseudo-first order relationship with kinetic rate constants of 0.015 min-1 for E1 and E2, 0.016 min-1 for E3 and 0.040 min-1 for EE2. The floc formed in-situ were characterized by determining the crystalline phases with X-ray diffraction, the size and zeta potential, and the shape and major components using scanning electron microscope with energy-dispersive X-ray spectrometer. The iron coagulant generated during electrocoagulation was lepidocrocite with a point of zero charge of 5.67 and an average floc diameter of 2255 nm.

Removal of trace metal contaminants from potable water by electrocoagulation.

This study investigated the effects of four operational and environmental variables on the removal of trace metal contaminants from drinking water by electrocoagulation (EC). Removal efficiencies for five metals (arsenic, cadmium, chromium, lead and nickel) were compared under varying combinations of electrode material, post-treatment, water composition and pH. Iron electrodes out-performed aluminum electrodes in removing chromium and arsenic. At pH 6.5, aluminum electrodes were slightly more effective at removing nickel and cadmium, while at pH 8.5, iron electrodes were more effective for these metals. Regardless of electrode, cadmium and nickel removal efficiencies were higher at pH 8.5 than at pH 6.5. Post-EC treatment using membrane filtration (0.45 µm) enhanced contaminant removal for all metals but nickel. With the exception of lead, all metals exhibited poorer removal efficiencies as the ionic strength of the background electrolyte increased, particularly in the very high-solids synthetic groundwaters. Residual aluminum concentrations were lowest at pH 6.5, while iron residuals were lowest in low ionic strength waters. Both aluminum and iron residuals required post-treatment filtration to meet drinking water standards. EC with post-treatment filtration appears to effectively remove trace metal contaminants to potable water standards, but both reactor and source water parameters critically impact removal efficiency.