Nobutaka Shirasaki

Comparison of behaviors of two surrogates for pathogenic waterborne viruses, bacteriophages Qβ and MS2, during the aluminum coagulation process.

Differences in the behaviors of two surrogates for pathogenic waterborne viruses, F-specific RNA bacteriophages Qbeta and MS2, were investigated during the coagulation process by using river water spiked with these bacteriophages. The particle size and electrophoretic mobility of Qbeta and MS2 were similar, but the removal performances of infectious Qbeta and MS2, as measured by a plaque forming unit (PFU) method, differed markedly during the coagulation process. The removal ratio of the infectious Qbeta concentration was approximately 2log higher than that of the infectious MS2 concentration at all coagulant doses tested. The total Qbeta and MS2 bacteriophage concentrations, which were measured by a real-time reverse transcription-polymerase chain reaction (RT-PCR) method and represented the total number of bacteriophages regardless of their infectivity, were similar after the coagulation process, suggesting that the behaviors of Qbeta and MS2 as particles were similar during the coagulation process. The difference between total concentration and infectious concentration indicated that some of the bacteriophages were probably inactivated during the coagulation process. This difference was larger for Qbeta than MS2, meaning that Qbeta was more sensitive to the virucidal activity of the aluminum coagulant. Analysis of the PFU and real-time RT-PCR findings together suggested that the difference in removal performances of Qbeta and MS2 during the coagulation process was probably caused by differences not in the extent of bacteriophage entrapment in the aluminum floc particles but in the sensitivity to virucidal activity of the aluminum coagulant.

Minimizing residual aluminum concentration in treated water by tailoring properties of polyaluminum coagulants

Aluminum coagulants are widely used in water treatment plants to remove turbidity and dissolved substances. However, because high aluminum concentrations in treated water are associated with increased turbidity and because aluminum exerts undeniable human health effects, its concentration should be controlled in water treatment plants, especially in plants that use aluminum coagulants. In this study, the effect of polyaluminum chloride (PACl) coagulant characteristics on dissolved residual aluminum concentrations after coagulation and filtration was investigated. The dissolved residual aluminum concentrations at a given coagulation pH differed among the PACls tested. Very-high-basicity PACl yielded low dissolved residual aluminum concentrations and higher natural organic matter (NOM) removal. The low residual aluminum concentrations were related to the low content of monomeric aluminum (Ala) in the PACl. Polymeric (Alb)/colloidal (Alc) ratio in PACl did not greatly influence residual aluminum concentration. The presence of sulfate in PACl contributed to lower residual aluminum concentration only when coagulation was performed at around pH 6.5 or lower. At a wide pH range (6.5-8.5), residual aluminum concentrations <0.02 mg/L were attained by tailoring PACl properties (Ala percentage ≤0.5%, basicity ≥85%). The dissolved residual aluminum concentrations did not increase with increasing the dosage of high-basicity PACl, but did increase with increasing the dosage of normal-basicity PACl. We inferred that increasing the basicity of PACl afforded lower dissolved residual aluminum concentrations partly because the high-basicity PACls could have a small percentage of Ala, which tends to form soluble aluminum-NOM complexes with molecular weights of 100 kDa-0.45 μm.

Estimation of norovirus removal performance in a coagulation-rapid sand filtration process by using recombinant norovirus VLPs

Norovirus (NV) is an important human pathogen that causes epidemic acute nonbacterial gastroenteritis worldwide. Because of the lack of a cell culture system or an animal model for this virus, studies of drinking water treatment such as separation and disinfection processes are still hampered. We successfully estimated NV removal performance during a coagulation-rapid sand filtration process by using recombinant NV virus-like particles (rNV-VLPs) morphologically and antigenically similar to native NV. The behaviors of two widely accepted surrogates for pathogenic waterborne viruses, bacteriophages Qbeta and MS2, were also investigated for comparison with that of rNV-VLPs. Approximately 3-log(10) removals were observed for rNV-VLPs with a dose of 40 muM-Al or -Fe, as polyaluminum chloride at pH 6.8 or ferric chloride at pH 5.8, respectively. Smaller removal ratios were obtained with alum and ferric chloride at pH 6.8. The removal performance for MS2 was somewhat larger than that for rNV-VLPs, meaning that MS2 is not recommended as an appropriate surrogate for native NV. By comparison, the removal performance for Qbeta was similar to, or smaller than, that for rNV-VLPs. However, the removal performances for rNV-VLPs and Qbeta differed between the coagulation process and the following rapid sand filtration process. Therefore, Qbeta also is not recommended as an appropriate surrogate for native NV.

Investigating norovirus removal by microfiltration, ultrafiltration, and precoagulation-microfiltration processes using recombinant norovirus virus-like particles and real-time immuno-PCR.

The removal of microorganisms by drinking water treatment processes has been widely investigated in laboratory-scale experiments using artificially propagated microorganisms. However, this approach cannot be applied to norovirus removal, because this virus does not grow in cell or organ culture, and this fact has hampered our ability to investigate its behavior during drinking water treatment. To overcome this difficulty, our research group previously used recombinant norovirus virus-like particles (rNV-VLPs), which consist of an artificially expressed norovirus capsid protein, in laboratory-scale drinking water treatment experiments. However, the enzyme-linked immunosorbent assay (ELISA) method generally used to detect rNV-VLPs is not sensitive enough to evaluate high removal ratios such as those obtained by ultrafiltration (UF). We therefore developed and applied a real-time immuno-polymerase chain reaction (iPCR) assay for rNV-VLP quantification to investigate norovirus removal by microfiltration (MF), UF, and hybrid precoagulation-MF processes. The rNV-VLP detection limit with the developed iPCR assay was improved at least 1000-fold compared with ELISA. Whereas MF with a nominal pore size of 0.1 μm could not eliminate NV-VLPs, a 4-log reduction was achieved by UF with a molecular weight cutoff of 1 kDa. When MF was combined with precoagulation (≥10 μmol-Fe/L for ferric chloride; ≥20 μmol-Al/L for polyaluminum chloride; ≥40 μmol-Al/L for alum), the performance of the hybrid process in eliminating rNV-VLPs was greater than that achieved by the 1 kDa UF. For all processes, the removal ratios of the bacteriophages MS2 and Qβ were greater than the rNV-VLP removal ratios by 1-2 logs, so neither bacteriophage can be recommended as a possible conservative surrogate for predicting the behavior of native NV during these processes.

Virus inactivation during coagulation with aluminum coagulants

We used the bacteriophages Qβ and MS2 to determine whether viruses are inactivated by aluminum coagulants during the coagulation process. We performed batch coagulation and filtration experiments with virus-containing solutions. After filtering the supernatant of the coagulated solution through a membrane with a pore size of 50 nm, we measured the virus concentration by both the plaque forming unit (PFU) and polymerase chain reaction (PCR) methods. The virus concentration determined by the PFU method, which determines the infectious virus concentration, was always lower than that determined by the PCR-based method, which determines total virus concentration, regardless of infectivity. This discrepancy can be explained by the formation of aggregates consisting of several virus particles or by the inactivation of viruses in the coagulation process. The former possibility can be discounted because (i) aggregates of several virus particles would not pass through the 50-nm pores of the filtration membrane, and (ii) our particle size measurements revealed that the virus particles in the membrane filtrate were monodispersed. These observations clearly showed that non-infectious Qβ particles were present in the membrane filtrate after the coagulation process with aluminum coagulants. We subsequently revealed that the viruses lost their infectivity after being mixed with hydrolyzing aluminum species during the coagulation process.

Adsorption capacities of activated carbons for geosmin and 2-methylisoborneol vary with activated carbon particle size: Effects of adsorbent and adsorbate characteristics

The adsorption capacities of nine activated carbons for geosmin and 2-methylisoborneol (MIB) were evaluated. For some carbons, adsorption capacity substantially increased when carbon particle diameter was decreased from a few tens of micrometers to a few micrometers, whereas for other carbons, the increase of adsorption capacity was small for MIB and moderate for geosmin. An increase of adsorption capacity was observed for other hydrophobic adsorbates besides geosmin and MIB, but not for hydrophilic adsorbates. The parameter values of a shell adsorption model describing the increase of adsorption capacity were negatively correlated with the oxygen content of the carbon among other characteristics. Low oxygen content indicated low hydrophilicity. The increase of adsorption capacity was related to the hydrophobic properties of both adsorbates and activated carbons. For adsorptive removal of hydrophobic micropollutants such as geosmin, it is therefore recommended that less-hydrophilic activated carbons, such as coconut-shell-based carbons, be microground to a particle diameter of a few micrometers to enhance their equilibrium adsorption capacity. In contrast, adsorption by hydrophilic carbons or adsorption of hydrophilic adsorbates occur in the inner pores, and therefore adsorption capacity is unchanged by particle size reduction.

Improved virus removal by high-basicity polyaluminum coagulants compared to commercially available aluminum-based coagulants

We investigated the effects of basicity, sulfate content, and aluminum hydrolyte species on the ability of polyaluminum chloride (PACl) coagulants to remove F-specific RNA bacteriophages from river water at a pH range of 6-8. An increase in PACl basicity from 1.5 to 2.1 and the absence of sulfate led to a reduction of the amount of monomeric aluminum species (i.e., an increase of the total amount of polymeric aluminum and colloidal aluminum species) in the PACl, to an increase in the colloid charge density of the PACl, or to both and, as a result, to high virus removal efficiency. The efficiency of virus removal at around pH 8 observed with PACl-2.1c, a nonsulfated high-basicity PACl (basicity 2.1-2.2) with a high colloidal aluminum content, was larger than that observed with PACl-2.1b, a nonsulfated high-basicity PACl (basicity 2.1-2.2) with a high polymeric aluminum content. In contrast, although extremely high basicity PACls (e.g., PACl-2.7ns, basicity 2.7) effectively removed turbidity and UV260-absorbing natural organic matter and resulted in a very low residual aluminum concentration, the virus removal ratio with PACl-2.7ns was smaller than the ratio with PACl-2.1c at around pH 8, possibly as a result of a reduction of the colloid charge density of the PACl as the basicity was increased from 2.1 to 2.7. Liquid (27)Al NMR analysis revealed that PACl-2.1c contained Al30 species, which was not the case for PACl-2.1b or PACl-2.7ns. This result suggests that Al30 species probably played a major role in virus removal during the coagulation process. In summary, PACl-2.1c, which has high colloidal aluminum content, contains Al30 species, and has a high colloid charge density, removed viruses more efficiently (>4 log10 for infectious viruses) than the other aluminum-based coagulants-including commercially available PACls (basicity 1.5-1.8), alum, and PACl-2.7ns-over the entire tested pH (6-8) and coagulant dosage (0.54-5.4 mg-Al/L) ranges.

Changes in mutagenicity and acute toxicity of solutions of iodinated X-ray contrast media during chlorination.

In the present study, the effects of chlorination on the mutagenicity (assessed via the Ames assay) and acute toxicity (assessed via a bioluminescence inhibition assay) of solutions containing one of five commonly used iodinated X-ray contrast media (ICM) (iopamidol, iohexol, iopromide, iomeprol, and diatrizoate) were investigated. Of the five ICM tested, only iopamidol was degraded by chlorine. Chlorination of the iopamidol-containing solution induced both mutagenicity and acute toxicity, which increased with chlorination time (up to 96 h). The areas of five out of 54 peaks detected on the LC/MS total ion chromatogram had good correlation (r(2)>0.90) between peak area and observed mutagenicity. To identify possible contributors to the observed mutagenicity, the Ames assay and LC/MS analysis were conducted on samples collected at 48-h chlorination time and extracted under different pH conditions. Of the five peaks, one peak was detected in the sample extracted at pH 7, but this sample was not mutagenic, indicating that the peak was not related to the observed mutagenicity. MS/MS analysis with an orbitrap mass spectrometer of the remaining four peaks revealed that two of the peaks represented the same TP (detected in negative and positive ion modes). Finally, three TPs were identified as suspected contributors to the mutagenicity induced by the iopamidol-containing solution after chlorination: 5-[(1,3-dihydroxypropan-2-yl)carbamoyl]-3-[(3-hydroxypropanoyl)oxy]-2,4-diiodobenzoic acid; N-(1,3-dihydroxypropan-2-yl)-3-(2,3-dioxopropyl)-2,4,6-triiodobenzamide; and 3-[(1,3-dihydroxypropan-2-yl)carbamoyl]-5-[(3-hydroxybutanoyl)oxy]-2,4,6-triiodobenzoic acid. Prediction of the mutagenicity potential of these three TPs with a battery of four quantitative structure-activity relationship models did not contradict our conclusion that these TPs contributed to the observed mutagenicity.

Isotope microscopy visualization of the adsorption profile of 2-methylisoborneol and geosmin in powdered activated carbon.

Decreasing the particle size of powdered activated carbon may enhance its equilibrium adsorption capacity for small molecules and micropollutants, such as 2-methylisoborneol (MIB) and geosmin, as well as for macromolecules and natural organic matter. Shell adsorption, in which adsorbates do not completely penetrate the adsorbent but instead preferentially adsorb near the outer surface of the adsorbent, may explain this enhancement in equilibrium adsorption capacity. Here, we used isotope microscopy and deuterium-doped MIB and geosmin to directly visualize the solid-phase adsorbate concentration profiles of MIB and geosmin in carbon particles. The deuterium/hydrogen ratio, which we used as an index of the solid-phase concentration of MIB and geosmin, was higher in the shell region than in the inner region of carbon particles. Solid-phase concentrations of MIB and geosmin obtained from the deuterium/hydrogen ratio roughly agreed with those predicted by shell adsorption model analyses of isotherm data. The direct visualization of the localization of micropollutant adsorbates in activated carbon particles provided direct evidence of shell adsorption.

Selecting pesticides for inclusion in drinking water quality guidelines on the basis of detection probability and ranking

Pesticides released into the environment may pose both ecological and human health risks. Governments set the regulations and guidelines for the allowable levels of active components of pesticides in various exposure sources, including drinking water. Several pesticide risk indicators have been developed using various methodologies, but such indicators are seldom used for the selection of pesticides to be included in national regulations and guidelines. The aim of the current study was to use risk indicators for the selection of pesticides to be included in regulations and guidelines. Twenty-four risk indicators were created, and a detection rate was defined to judge which indicators were the best for selection. The combination of two indicators (local sales of a pesticide for the purposes of either rice farming or other farming, divided by the guideline value and annual precipitation, and amended with the scores from the physical and chemical properties of the pesticide) gave the highest detection rates. In this case study, this procedure was used to evaluate 134 pesticides that are currently unregulated in the Japanese Drinking Water Quality Guidelines, from which 44 were selected as pesticides to be added to the primary group in the guidelines. The detection probability of the 44 pesticides was more than 72%. Among the 102 pesticides currently in the primary group, 17 were selected for withdrawal from the group.