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Abstract During runoff process, the urban road dust (URD) passes through the road-side gutters and detention tanks, where it gets hold for a certain period of time, hours to weeks, before making its way into the water bodies. A part of the water-exchangeable toxicants are leached by the water, and some strongly bound toxicants remain attached to the URD. Toxicity of urban runoff is dependent on holding time, water volume, and the type and composition of the wet road dust (WeRD) and leachate. However, there are no studies that have elucidated the manner in which toxicities of the WeRD and leachate vary during prolonged holding in the runoff processes. The main objectives of this research were to, i) identify the distribution of toxicants in the WeRD and leachate, and ii) evaluate the change in toxicity during prolonged holding. The URD samples that were collected from residential road, arterial road and highways in Tokyo, Japan, were mixed with moderately hard water in varying ratios (1:2 to 1:16) and were held for a certain time (1 h to 5 days) before centrifuging into the WeRD and leachate. Toxicity test was conducted with ostracod Heterocypris incongruens direct contact test. Toxicity of both the WeRD and leachate from residential area was not significantly greater than the 20% lethal limit. Toxicity of the WeRD from other stations initially decreased, further reached a minimum corresponding to the critical holding time, and it subsequently increased again. Toxicity of the arterial leachate gradually increased, whereas that of the highway leachate gradually decreased during 5 days holding. Unionized ammonia and zinc were confirmed as one of the possible toxicants. This study proposes and verifies a model for the toxicity change of the WeRD during prolonged holding. Graphical abstract Unlabelled Image Highlights • Toxicity change of road dust during runoff process is elucidated. • Toxicity of wet road dust (WeRD) decreased initially and gradually increased. • Unionized ammonia and Zinc were confirmed as one of the toxicants. • Model for the toxicity change of the WeRD during prolonged holding is proposed and verified. [ABSTRACT FROM AUTHOR]

Biological filters effectively remove ammonium from drinking water via nitrification. In a pilot-scale upflow biological contact filter (U-BCF), complete ammonia oxidizers (comammox), which are capable of oxidizing ammonia to nitrate in one cell, were more abundant than ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). However, little information is available on the contribution of comammox to nitrification. In this study, we evaluated the autotrophic growth activity of comammox associated with biological activated carbon (BAC) in a U-BCF by DNA-stable isotope probing (DNA-SIP). BAC samples collected from the U-BCF were continuously fed mineral medium containing 0.14 mg N L−1 ammonium and 12C- or 13C-labeled bicarbonate for 20 days. DNA-SIP analysis revealed that comammox (clades A and B) as well as AOA assimilated bicarbonate after 10 days of incubation, proving that dominant comammox could contribute to nitrification. Contrarily, AOB remained inactive throughout the observation period. Amplicon sequencing of the 13C-labeled DNA fractions of comammox revealed that specific genotypes other than the most dominant genotype in the original sample were more enriched under the incubation condition for the DNA-SIP experiment. Thus, dominant genotypes of comammox in a U-BCF might utilize organic nitrogen to fuel nitrification in ammonia-limited environments. [ABSTRACT FROM AUTHOR]

The current study involves characterization of organic compounds, heavy metals, and ammonia as potential toxicants in one arterial road (St. 3) and two highway (St. 7 and 8) urban road dust (URD) collected in Tokyo, Japan. URD toxicity was evaluated by Toxicity Identification Evaluation using the ostracod Heterocypris incongruens direct contact test. URDs were treated with resins (10% XAD-4, 20% SIR-300 and 20% SIR-600) to determine the reduction in mortality after treatment. The mortality of ostracods exposed to St. 3 URD (baseline 80%) was significantly (p < 0.05) reduced to 0% and 47% after XAD-4 and SIR-300 treatment respectively. This reduction led us to suspect hydrophobic organic compounds and heavy metals as potential toxicants. Subsequent elution of the recovered XAD-4 with polar (methanol, acetone) and non-polar (dichloromethane) solvents confirmed the dominance of relatively polar hydrophobic organic toxicants. The dissolved concentration of Cu and Zn after SIR-300 treatment exceeded the 50% lethal concentration (LC50) for ostracods, which led us to suspect Cu and Zn as other probable toxicants. The concentration of Zn in a SIR-300 acidic elutriate, recovered after the termination of toxicity test, confirmed Zn as one of the toxicants in St. 3. The baseline mortality (100%) of St. 7 was significantly reduced (23%) by SIR-300 treatment. This indicated the presence of heavy metals as the probable toxicant. However, the concentration of dissolved heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in the overlying water was below previously reported LC50 values for each metal in St. 7. Recovery of Zn concentrations exceeding the LC50 in the St. 7 SIR-300 elutriate led us to suspect a dietary exposure route of Zn to the ostracod during the direct contact test. The overall results indicate that the toxicant types can vary widely depending on the road sampled. [ABSTRACT FROM AUTHOR]

Highlights: [•] Toxicity is discussed based on dose–response, size-fractions and holding time. [•] Standardized heavy metal and PAH concentration is applied to differentiate toxicity. [•] Median lethal concentration (LC50) of urban road dust varied (v/v) from 3.8 to 67%. [•] Fraction of fine particles were not always more toxic than the other fractions. [•] Toxicity of urban road dust was site specific. [Copyright &y& Elsevier]

Abstract: Soakaway sediment trapped in urban infiltration facilities exhibits high heavy metal content that makes it an important diffuse pollution source of groundwater contamination. In this study, heavy metal (Cu, Zn, Cd and Pb) content, speciation and stable isotopic variation of Pb in soakaway sediments were coupled and compared with soils (surface and underlying), and road dusts (residential and highway) to trace their contamination characteristics, mobility and source. Soakaway sediment and residential road dust were found to exhibit higher metallic content of anthropogenic origin with high mobility. Isotopic ratios in exchangeable fraction showed the least 206Pb/207Pb and 208Pb/207Pb ratios. Although there were significant differences in total metal content and speciation between two road dust samples, their isotopic signatures exhibited close resemblance and aligned with anthropogenic markers such as paint, petrol and aerosols. Heterogeneity of isotopic signature in different fractions of soakaway sediments seems to have originated from the mixing of multiple sources of heavy metals. Study concludes that mobility, source and contamination characteristics of metals in soakaway sediment were different from urban soil and road dust samples due to their complex environment and regular metal supply by road runoff. [Copyright &y& Elsevier]

Twelve particle-bound polycyclic aromatic hydrocarbons (PAHs) were measured in the first flush regime of road runoff during nine events in Winterthur in Switzerland. The total PAH contents ranged from 17 to 62 μg/g. The PAH patterns measured at different time intervals during the first flush periods were very similar within each event irrespective of variation in suspended solids (SS) concentration within the first flush regime. However, the PAH patterns were different from event to event. This indicates that the environment plays an important role in PAH accumulation in SS. A toxicity identification evaluation approach using a toxicity equivalency factor (TEF) was applied to compare toxicities in the different events. The TEFs were found to be between 8 and 33 μg TEQ g-1 (TEQ: toxic equivalent concentration). In some cases, two events having similar total PAH contents showed two fold toxicity differences. [ABSTRACT FROM AUTHOR]

Abstract: Artificial infiltration facilities (AIFs) that enhance groundwater recharge and regulate urban runoff are going to be an integral element of the urban infrastructure. However, AIFs provide a sink which trap pollutants that are likely to cause groundwater contamination. The current study aimed first to examine the mobility characteristics of Cu, Zn, and Pb through soil and soakaway sediment using an integrated analytical approach for column leaching with artificial road runoff (ARR) and then to differentiate the sorption patterns among different samples, (i.e., surface soil, underlying soil, and soakaway sediments) using mass balances. In addition, the study compares metal retention and release under continuous and intermittent flow conditions. Column leaching experiments were conducted using batches for 10 and 30d under continuous flow condition and for 20d of intermittent leaching. Heavy metal content and partitioning in soil and sediment used in columns were well characterized before and after leaching experiments. The results showed that a gradual increase in pH and decrease in dissolved organic carbon had pronounced effect on the mobilization of heavy metals. Pb showed the highest retention compared to Cu and Zn which implies that metal complexes play a pivotal role in metal transport. Labile fractions were found to be trapped by the solid materials for retention and their high concentration in ARR is a major concern from the pollution point of view through infiltration facilities. Results obtained in this study predict the risk associated with the release of retained heavy metal under changing environmental conditions in AIFs. [Copyright &y& Elsevier]

Selective sequential dissolution (SSD) and isotopic dilution are two most commonly applied techniques for the measurement of mobile fraction of heavy metal present in the urban environment. This work examined the compliance between SSD proposed by the Community Bureau of Reference (BCR) and isotopic dilution technique (IDT) for determination of mobile pool of heavy metal contained in soakaway sediment, road dust, and soil sample. Heavy metals (Cu, Zn, Cd and Pb) were fractionated using the three-stage BCR protocol, while isotopically exchangeable metal concentrations (E-value) were investigated through isotopic tracers (111Cd, 65Cu, 207Pb and 66Zn). In general, total contamination level, E-value and BCR exchangeable fractions of different samples followed the similar order of road dust . soakaway sediment . soil. Results revealed that the E-value exceeded the BCR exchangeable fraction in all samples. In addition, the first three fractions of BCR which have potential to become mobile under certain environmental conditions were collectively termed as "potential mobile pool" and compared with E-value. It was concluded that metal extracted by weak acid underestimates the exchangeable fraction while the potential mobile pool extracted by three reagents overestimates the real mobile forms of heavy metals. However, better mobility characteristics of heavy metals can be assessed by coupling information obtained through sequential extraction and isotopic exchange. [ABSTRACT FROM AUTHOR]

Lead (Pb) isotopic data were used in this study to first distinguish the partitioning of anthropogenic and natural lead in different fractions, obtained by BCR sequential extraction, and then to anticipate their mixing process in the soakaway sediment of artificial infiltration facilities (AIF). Total metal content was found higher in soakaway sediment samples than that of soil. The lowest 206Pb/207Pb ratios were mostly observed in exchangeable fractions of soil and sediment samples, while residual fractions mostly showed the highest 206Pb/207Pb and 208Pb/207Pb ratios than those of other fractions. In general, both ratios were higher in the soil than those of sediments. Further among soil samples, residual fraction of bottom soil exhibited higher ratios than surface soil indicating higher contribution of natural lead with depth. In addition, the difference in Pb content, partitioning and its isotope signature among four sediment samples were also investigated considering their sampling locations. The plot of 206Pb/207Pb versus 208Pb/207Pb showed two well demarcated cluster formations by soil and sediments samples that describe the partitioning between anthropogenic and natural lead; and some points falling in between soil and sediment samples pertinently illustrated the mixing processes between these two different pools of lead. [ABSTRACT FROM AUTHOR]

Abstract: Concern about the sustainability of urban water use is the strong motivation to understand the potential of rainwater use and water recycling in urbanized cities. The history of water supply in Tokyo and its experience may provide useful information to develop sustainable urban water use and find future possible tasks in rapidly growing cities. Besides, various innovative strategies to meet the current and future water demand in Tokyo may help us to consider new approaches adjusting to the developing mega cities in Asia. In this paper, the past and current practices on utilization of latent water resources such as rainwater and reclaimed wastewater in Tokyo are summarized from the viewpoint of sustainable water use. The storage of rainwater is a useful measure for water demand in emergency cases. In addition, the rainwater use can work as a kind of environmental education to make citizens aware of sustainable urban water use. There are 850 facilities for rainwater use in Tokyo. Since reclaimed wastewater use has several benefits, a huge water volume has been utilized for various purposes such as washing, water-cooling, toilet flushing, waterway restoration and creation of recreational waterfront. From the viewpoint of human health risk, new micropollutants such as estrogens, endocrine disrupters and surfactants should be considered as quality guideline parameter besides the conventional ones. Importance of infiltration facilities should be also highlighted to secure the sound water cycle. Groundwater recharge through the infiltration facilities provide a potential storage of water resource which can be withdrawn in the future if necessary. [Copyright &y& Elsevier]

The aim of this paper is to introduce the concept of sustainable urban wastewater management in the Asian context. Sewerage systems are key facilities to support public health and sound development in urban areas. They exist in most of the rapidly developing cities of Asia; however, a range of practical, financial. political, and environmental factors mean that provision is often inadequate to meet current and projected demand. To meet clean water goals and reduce the environmental impacts of urbanization, sewerage systems should be incorporated property into watershed management plans. The paper ends by examining some of the range of new and established technologies and methods that can help Asian cities and periurban areas to minimize the burden and maximize the potential benefits of urban wastewater. [ABSTRACT FROM AUTHOR]

This study was aimed at investigating source apportionment of PAHs in urban road dust. Seven kinds of PAHs sources were defined: diesel vehicle exhaust, gasoline vehicle exhaust, tire, asphalt-pavement, asphalt or bitumen, petroleum products excluding tire and asphalt, and the combustion products except for those in vehicle engines. Using cluster analysis combined with principal component analysis, 189 source data were classified into 11 source groups (S1–S11) based on the content percentage of 12 individual PAHs (12-PAH profiles). Thirty-seven dust samples on nine streets in Tokyo were collected and analyzed for 12-PAH profiles. In order to estimate the PAHs, contributions of S1–S11 to dust samples, the multiple regression analysis was performed. The 12-PAH profiles of each dust sample and those of 11 source groups were applied as dependent and independent variables, respectively. We defined the comparative contribution of each source group as the ratio of each regression coefficient to the sum of them. The result revealed that diesel vehicle exhaust, tire, and pavement were the major contributors of PAHs in the road dust. [ABSTRACT FROM AUTHOR]

The primary concentration and molecular process are critical to implement wastewater-based epidemiology for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the previously developed methods were optimized for nonenveloped viruses. Few studies evaluated if the methods are applicable to the efficient recovery of enveloped viruses from various types of raw sewage. This study aims (1) to compare the whole process recovery of Pseudomonas phage φ6, a surrogate for enveloped viruses, among combinations of primary concentration [ultrafiltration (UF), electronegative membrane vortex (EMV), and polyethylene glycol precipitation (PEG)] and RNA extraction methods (spin column-based method using QIAamp Viral RNA Mini Kit and acid guanidinium thiocyanate–phenol–chloroform extraction using TRIzol reagent) for three types of raw sewage and (2) to test the applicability of the method providing the highest φ6 recovery to the detection of SARS-CoV-2 RNA. Among the tested combinations, PEG+TRIzol provided the highest φ6 recovery ratio of 29.8% to 49.8% (geometric mean). UF + QIAamp Viral RNA Mini Kit provided the second highest φ6 recovery of 6.4% to 35.8%. The comparable φ6 recovery was observed for UF + TRIzol (13.8–30.0%). PEG + QIAamp Viral RNA Mini Kit provided only 1.4% to 3.0% of φ6 recovery, while coliphage MS2, a surrogate for nonenveloped viruses, was recovered comparably with PEG + TRIzol. This indicated that the nonenveloped surrogate (MS2) did not necessarily validate the efficient recovery for enveloped viruses. EMV + QIAamp Viral RNA Mini Kit provided significantly different φ6 recovery (1.6–21%) among the types of raw sewage. Then, the applicability of modified PEG + TRIzol was examined for the raw sewage collected in Tokyo, Japan. Of the 12 grab samples, 4 were positive for SARS-CoV-2 CDC N1 and N3 assay. Consequently, PEG + TRIzol provided the highest φ6 recovery and allowed for the detection of SARS-CoV-2 RNA from raw sewage. Unlabelled Image • The combinations of primary concentration and RNA extraction were compared. • PEG + TRIzol provided the highest φ6 recovery ranging from 29.8–49.8%. • Nonenveloped surrogates did not necessarily validate the efficient recovery of φ6. • SARS-CoV-2 RNA was detected by modified PEG+TRIzol from raw sewage in Japan. [ABSTRACT FROM AUTHOR]

Isotope dilution techniques (IDT) and sequential extraction procedures (SEPs) were compared to apprehend the differences between two techniques in determining metal exchangeability and vulnerability to pollute the urban groundwater. For this purpose, soil (n = 2), "soakaway" sediment deposited in the artificial infiltration facilities (AIF) (n = 4), and road dust (n = 2) were sampled from Tokyo metropolitan. Sorption coefficients of four metals (Cu, Zn, Cd and Pb) were assessed through isotopic exchangeability (E-value) and potential mobile pool i.e. addition of exchangeable, reducible and oxidizable fraction obtained by Community Bureau of Reference (BCR)-procedures. The E-value for the three samples were found smaller than the potential mobile pool but were higher than BCR-exchangeable fractions. The use of strong extractants are likely to play an active role in the disagreement between SEPs and IDT. IDT accounts for the isotopic exchangeability while BCR provides information of vulnerability of metals associated with different fractions that can leach under different environmental conditions. Sorption coefficients measured in soakaway sediment was found comparable to soil thus likely to retain metals. However, as variability in environmental conditions is likely to affect Kd, the soakaway sediment may become an active metal source in future rather than acting as the permanent sink. The study concludes that there is the possibility of errors while predicting metal vulnerability to groundwater with both techniques and thus a model compliance integrating the virtue of both techniques will be a way forward. Image 1 • Better compliance amid isotope dilution and sequential extraction for high metal loading. • Isotopic exchangeability (E-value) was smaller than extracted potential mobile pool. • Coupling of IDT with BCR provide better estimate of leaching vulnerability for metals. • IDT accounts for actual exchangeability while SEPs provide leaching vulnerability. [ABSTRACT FROM AUTHOR]

Assays based on the polymerase chain reaction (PCR) are widely applied to quantify enteric viruses in aquatic environments to study their fates and potential infection risks. However, inhibitory substances enriched by virus concentration processes can result in inaccurate quantification. This study aimed to find a method for improving virus quantification by mitigating the effects of inhibitory environmental concentrates, using previous knowledge of the properties of the inhibitory substances. Performances of anion exchange resins, gel filtration, and a hydrophobic resin (DAX-8) were comparatively evaluated using poliovirus and its extracted RNA spiked into humic acid solutions. These solutions served as good representatives of the inhibitory environmental concentrates. A sequential treatment using DAX-8 resin and gel filtration produced the most favorable results, i.e., low virus losses that were stable and a reduced inhibitory effect. Furthermore, the sequential treatment was applied to another set of 15 environmental concentrates. Without the sequential treatment, serious underestimation (>4.0 log 10 to 1.1 log 10) of a molecular process control (murine norovirus) was measured for eight samples. With the treatment, the control was detected with <1.0 log 10 underestimation for all samples. The treatment improved the quantification of seven types of indigenous viruses. In summary, the sequential treatment is effective in improving the viral quantification in various of environmental concentrates. Image 1 • Some organic substances hinder virus detection after concentration processes. • Various treatments to mitigate these inhibitory effects were tested. • Performance of a treatment using DAX-8 resin with gel filtration was satisfactory. • The treatment improved the quantification of variable viruses in environmental samples. [ABSTRACT FROM AUTHOR]

This study was aimed at comparing the Zn and Cu binding parameters with dissolved organic matter (DOM) in road runoff, wastewater treatment plant (WWTP) effluent and influent. Conditional stability constant (K') and binding site concentration ([L]) are important to predict free or labile metal concentration and toxicity in the water environment. The values of K' and [L] of three DOMs were determined by metal titration and Scatchard linearization. The Zn and Cu titration data for DOMs in WWTP effluent and influent fitted to a two-ligand model, while DOM in road runoff fitted to a single-ligand model. The order of the values of K' was WWTP influent > WWTP effluent > road runoff both for Zn and Cu. Total ambient binding site concentration ([LT]) was lower in DOM of road runoff (1.4-1.5μM) than those in DOM of WWTP effluent (1.6-2.3μM) and influent (17-18μM), suggesting lower metal complexation capacity in DOM of road runoff. DOM in WWTP influent was expected to bind to both Zn and Cu more strongly than that of the effluent DOM. [ABSTRACT FROM AUTHOR]

Abstract: Groundwater contamination by heavy metals from infiltration facilities receiving road runoff is of potential concern. In this study, sorption tests were conducted to evaluate the influence of the water quality of road runoff, especially dissolved organic matter (DOM), on the sorption of heavy metal species by soakaway sediment. Sequential batch tests were conducted to assess metal sorption by the soakaway sediment receiving road runoff from residential and heavily trafficked areas. Ni was adsorbed by the sediment, indicating that soakaway sediments function to prevent groundwater contamination by Ni. In contrast, Zn was released from the soakaway sediment in sorption tests using heavily trafficked road dust leachates. Ni, Cu, Zn, and dissolved organic carbon concentrations were higher in soakaway sediment leachates obtained by sorption tests using heavily trafficked road dust leachates than those using residential road dust leachates, suggesting traffic activities contaminate these pollutants. A large portion of Zn, released from the soakaway sediment, existed as stable complexes. DOM in road runoff possibly enhances the release of Zn from the sediments within infiltration facilities and might cause groundwater contamination. [Copyright &y& Elsevier]

Infiltration facilities are designed for both the retention of non-point pollutants and the replenishment of groundwater in urban areas. In this study, sorption tests were conducted to evaluate the speciation of heavy metals and their behaviour in infiltration facilities receiving urban road runoff containing high DOC concentrations and stable heavy metal organic complexes. Road dust and three soakaway sediments were collected from heavy traffic areas and a residential area with an infiltration-type sewerage system in Tokyo, Japan. Sequential multiple batch tests were conducted by adding prepared road dust leachate (artificial road runoff) or deionised water to soakaway sediment to obtain soakaway sediment leachate (artificial percolating water from soakaway sediment), which mimicked the sorption by sediments in soakaways receiving urban road runoff. Heavy metal speciation was assessed by means of a combination of anion-exchange resin measurements and MINTEQA2 model calculations, and further validated by chelating resin measurements. In road dust leachates and soakaway sediment leachates, Cu predominantly existed as organic complexes and carbonates, whereas most Mn, Zn and Cd were found to exist in the form of free ions and carbonate complexes. Stable organic complexes of Cu in road dust leachates were strongly adsorbed by soakaway sediments despite the limited adsorption of DOC. On the other hand, desorption of free Mn, Zn and Cd ions from the sediment receiving road dust leachates was observed, indicating that heavy metals such as Mn, Zn and Cd may ultimately reach groundwater as free ions. [Copyright &y& Elsevier]

Catechol 2,3-dioxygenase (C23O) is an extradiol dioxygenase that plays an important role in degrading aromatic compounds such as those found at polluted sites. However, little is known about the diversity of C23O genes in unpolluted environments. In such environments, various factors, including the quality and quantity of dissolved organic matter (DOM), could influence the composition and behaviour of bacterial community possessing C230 genes. We investigated C23O genes in bacteria responding to DOM from various sources in a eutrophic lake by PCR and cloning. Six microcosms filled with lake water containing indigenous bacteria and DOM from different sources were incubated for 10 days. After 1 or 2 days of incubation, C23O genes were detected in the microcosms enriched with DOM recovered from inflow river water and humus from reed grass. The sequences were very diverse but had features conserved in extradiol dioxygenases. The clone libraries generated on day 2 showed distinctive compositions among microcosms, indicating that bacteria possessing a variety of C23O genes responded differently to DOM from different sources. After 10 days of incubation, C23O genes in a previously unidentified gene cluster, ‘Cluster X’, became dominant in the libraries. [ABSTRACT FROM AUTHOR]

Individual particles containing Cr and/or Pb and other major components were identified in road dust from a heavily used road (hereinafter ‘heavy traffic road dust’), road dust from a residential area and soakaway sediment by electron probe microanalyser to locate their sources and carrier particles. Individual particles containing high levels of Cr and/or Pb (⩾0. 2%) were identified using wavelength dispersive spectrometry (WDS) map analysis. Chromium, Pb and other major elements were then determined by means of a combination of WDS and energy-dispersive spectrometry in all identified particles, 50 particles containing neither Cr nor Pb from each type of road dust and soakaway sediment, and yellow road line markings. WDS map analysis revealed that many particles containing both Cr and Pb were present among the identified particles in heavy traffic road dust, whereas they were minor components in road dust from the residential area and soakaway sediment. The plots of X-ray intensities of Cr vs. Pb were linear for the identified particles containing both Cr and Pb in heavy traffic road dust, and the line closely fitted the plots for the three yellow road line marking samples. Individual particles were then classified using cluster analysis of element components. The results revealed that the adsorption of source materials or released metals onto soil minerals occurred in road dust and soakaway sediment, that the yellow road line markings were sources of Cr and Pb in heavy traffic road dust, and that materials containing Fe as a major component, such as stainless steel, were additional sources of Cr in both road dust and soakaway sediment. [Copyright &y& Elsevier]

Abstract: Polycyclic aromatic hydrocarbons (PAHs) present in size- and density-fractionated road dust were measured to identify the important fractions in urban runoff and to analyse their sources. Road dust was collected from a residential area (Shakujii) and a heavy traffic area (Hongo Street). The sampling of road dust from the residential area was conducted twice in different seasons (autumn and winter). The collected road dust was separated into three or four size-fractions and further fractionated into light (<1.7g/cm3) and heavy (>1.7g/cm3) fractions by using cesium chloride solution. Light particles constituted only 4.0±1.4%, 0.69±0.03% and 3.4±1.0% of the road dust by weight for Shakujii (November), Shakujii (February) and Hongo Street, respectively but contained 28±10%, 33±3% and 44±8% of the total PAHs, respectively. The PAH contents in the light fractions were 1–2 orders of magnitude higher than those in the heavy fractions. In the light fractions, the 12PAH contents in February were significantly higher than the 12PAH contents in November (P <0.01), whereas in the heavy fractions, no significant difference was found (P >0.05). Cluster analysis revealed that there was a significant difference in the PAH profiles between locations rather than between size-fractions, density-fractions and sampling times. Multiple regression analysis indicated that asphalt/pavement was the major source of Shakujii road dust, and that tyre and diesel vehicle exhaust were the major sources of finer and coarser fractions collected from Hongo Street road dust, respectively. [Copyright &y& Elsevier]

In this study, coagulation experiments were conducted using three coagulants (ferric chloride (FC), poly ferric sulfate (PFS) and poly aluminum chloride (PAC)) to evaluate DOM (dissolved organic matter) removal performance in lake water. DOM removal was characterized not only by comprehensive indices such as dissolved organic carbon (DOC) and UV absorbance at 260 nm (U VA260) but also by spectrofluorimetry and pyrolysis gas chromatography/mass spectrometry (py-GC/Ms) analysis, which are useful techniques to obtain information on the structure of organic matter. At a coagulant concentration of 0.3 mM, a higher percentage of DOC was removed by ferric coagulants than by the aluminum coagulant (FC: 53%, PFS: 41%, PAC: 36%). Similar trends were observed in the coagulation-induced decrease of UVA260 and trihalomethane formation potential (THMFP). Spectrofluorimetry showed that higher concentrations of coagulant increased the removal of DOM with fluorescence components. However, fluorescence intensity was not reduced by increased coagulant concentrations at emission wavelengths shorter than 350 nm. Treatments with ferric coagulants were more effective in removing fluorescent components than PAC. Comparison of pyrochromatograms before and after coagulation revealed that the peak area of propanoic acid decreased most significantly after coagulation by FC among 16 identified fragment compounds. Four fragment compounds differed greatly in their coagulation behaviour, depending on the coagulant used. [ABSTRACT FROM AUTHOR]

Abstract: Road and roof dust was collected and samples of runoff were taken at an urban storm sewer system in a residential area in Japan. Suspended solids (SS) in the runoff samples were classified into two fractions: fine (smaller than 45μm) and coarse (larger than 45μm). Runoff monitoring and chemical analysis data were also used to validate a runoff model for particle-bound polycyclic aromatic hydrocarbons (PAHs) that was originally developed to explain the behaviour of SS in the same area. The model, in which roads and roofs were considered separately as impervious surfaces, expressed the SS and particle-bound PAHs runoff behaviour for fine and coarse particles very well, except during and after heavy rainfalls (more than 10mm/h). However, the model could not explain the PAH profiles of runoff particles; the profiles of 12 PAH compounds tracked in this study were almost constant and more similar to those of road dust than roof dust throughout the event. An improved model is developed which explains the runoff behaviour by considering two types of road dust with different mobility. [Copyright &y& Elsevier]

• Capsid intergrity (RT-)qPCR was used to assess intact viruses in tap water • Intact pepper mild mottle virus (PMMoV) was detected in tap water • Aichivirus with damaged capsids was detected in tap water • Absence of intact PMMoV can be used to ensure the viral safety of tap water from surface water Capsid integrity (RT-)qPCR has recently been developed to discriminate between intact forms from inactivated forms of viruses, but its applicability to identifying integrity of viruses in drinking water has remained limited. In this study, we investigated the application of capsid integrity (RT-)qPCR using cis-dichlorodiammineplatinum (CDDP) with sodium deoxycholate (SD) pretreatment (SD-CDDP-(RT-)qPCR) to detect intact viruses in surface water and tap water. A total of 63 water samples (surface water, n = 20; tap water, n = 43) were collected in the Kanto region in Japan and quantified by conventional (RT)-qPCR and SD-CDDP-(RT-)qPCR for pepper mild mottle virus (PMMoV) and seven other viruses pathogenic to humans (Aichivirus (AiV), noroviruses of genotypes I and II, enterovirus, adenovirus type 40 and 41, and JC and BK polyomaviruses). In surface water, PMMoV (100%) was more frequently detected than other human pathogenic viruses (30%–60%), as determined by conventional (RT-)qPCR. SD-CDDP-(RT-)qPCR also revealed that intact PMMoV (95%) was more common than intact human pathogenic viruses (20%–45%). In the tap water samples, most of the target viruses were not detected by conventional (RT-)qPCR, except for PMMoV (9%) and AiV (5%). PMMoV remained positive (5%), whereas no AiV was detected when tested by SD-CDDP-(RT-)qPCR, indicating that some PMMoV had an intact capsid, whereas AiV had damaged capsids. The presence of AiV in the absence of PMMoV in tap water produced from groundwater may demonstrate the limitation of PMMoV as a viral indicator in groundwater. In addition to being abundant in surface water, PMMoV was detected in tap water, including PMMoV with intact capsids. Thus, the absence of intact PMMoV may be used to guarantee the viral safety of tap water produced from surface water. Image, graphical abstract [ABSTRACT FROM AUTHOR]

We investigated semiquantitative changes in almost 1000 dissolved organic matter (DOM) features during oxidation with 1 mg of O 3 per liter (mg O 3 /L), 4 mg O 3 /L, or 4 mg O 3 /L + 2.5 mg of H 2 O 2 per liter (advanced oxidation process, AOP) by unknown screening analysis with Orbitrap mass spectrometry. The consequential effects on formation of unknown disinfection by-products (DBPs) by chlorination were evaluated in laboratory-scale experiments. Several hundred unsaturated DOM features with positive oxygen-subtracted double bond equivalents per carbon ((DBE–O)/C) were decomposed by the ozone-only treatment and AOP. The AOP decomposed some saturated (negative (DBE–O)/C)) and reduced molecules, which had negative carbon oxidation states (C os). Several hundred saturated oxidation by-products were detected after ozonation and the AOP. After chlorination, the samples pre-treated with ozone alone resulted in higher formation of unknown DBPs than the AOP pre-treated sample or the sample without oxidation. Over half of the DBP precursors, estimated by electrophilic substitution, were not totally decomposed by any oxidation process, but they were increased after the ozone-only process and AOP. DBP precursors produced by the ozone-only process or AOP formed unique unknown DBPs. Therefore, post-treatment processes after oxidation and before chlorination are important to minimize formation of unknown DBPs. Image 1 • Changes of over 1000 DOM features by ozonation and peroxone were investigated. • Unsaturated DOM features were selectively decomposed by ozone and peroxone. • Several hundred of saturated oxidation byproducts were detected after ozonation and peroxone. • Ozone results in formation of unknown disinfection by-products more than peroxone. • Ozone and peroxone increase generate their specific unknown DBP precursors. [ABSTRACT FROM AUTHOR]

Highlights • Ferrihydrite (Fh) treatment was developed and optimized. • Fh treatment was able to effectively mitigate inhibition of RT-qPCR virus detection in humic acids and seawater concentrates. • Its recovery of viruses in heavily inhibited water samples ranged from 2% to 12%. • Fh treatment improved RT-qPCR detection of indigenous virus in seawater samples. Abstract Molecular assays are currently the most widely used method to quantify pathogenic viruses in water; however, their performance is often disrupted by matrices present in environmental samples. The present study used ferrihydrite (Fh) treatment to mitigate inhibition of RT-qPCR virus detection from environmental water concentrates. Fh treatment was performed to improve the detection of spiked Aichi virus 1 (AiV) and Qβ bacteriophage (Qβ) in commercial humic acids and seawater concentrates. The optimal Fh doses were found to be 200, 500, and 1000 mgFe/L for humic acid concentrates at UV 254 of 1.5, 3.0, and 6.0 cm−1 respectively, whereas this value was 1000 mgFe/L for the seawater concentrates at UV 254 of 1.5 cm−1. At these optimal doses, the recoveries of spiked AiV and Qβ ranged from 2% to 12% in the humic acid and seawater concentrates, respectively. In addition, high levels of indigenous viruses (including AiV ≥ 2.81 log 10 copies/mL and pepper mild mottle virus ≥ 2.77 log 10 copies/mL) were detected from seawater concentrates after Fh treatment, while none were detected without Fh treatment. Fh treatment effectively mitigated the inhibitive effects from environmental water samples. [ABSTRACT FROM AUTHOR]

To assess the potential of pepper mild mottle virus (PMMoV) as a viral process indicator, its reduction through coagulation–sedimentation (CS) and rapid sand filtration (RSF) were compared with those of Escherichia coli , previously used viral indicators, and norovirus genotype II (NoV GII; enteric virus reference pathogen) in a bench-scale experiment. PMMoV log 10 reductions in CS (1.96 ± 0.30) and RSF (0.26 ± 0.38) were similar to those of NoV GII (1.86 ± 0.61 and 0.28 ± 0.46). PMMoV, the most abundant viruses in the raw water, was also determined during CS, RSF, and advanced treatment processes at two full-scale drinking water treatment plants under strict turbidity management over a 13-month period. PMMoV was concentrated from large-volume water samples (10–614 L) and quantified by Taqman-based quantitative polymerase chain reaction. The PMMoV log 10 reduction in CS (2.38 ± 0.74, n = 13 and 2.63 ± 0.76, n = 10 each for Plant A and B) and in ozonation (1.91 ± 1.18, n = 5, Plant A) greatly contributed to the overall log 10 reduction. Our results suggest that PMMoV can act as a useful treatment process indicator of enteric viruses and can be used to monitor the log 10 reduction of individual treatment processes at drinking water treatment plants due to its high and consistent copy numbers in source water. [ABSTRACT FROM AUTHOR]

While the practice of rainwater harvesting (RWH) can be traced back millennia, the degree of its modern implementation varies greatly across the world, often with systems that do not maximize potential benefits. With a global focus, the pertinent practical, theoretical and social aspects of RWH are reviewed in order to ascertain the state of the art. Avenues for future research are also identified. A major finding is that the degree of RWH systems implementation and the technology selection are strongly influenced by economic constraints and local regulations. Moreover, despite design protocols having been set up in many countries, recommendations are still often organized only with the objective of conserving water without considering other potential benefits associated with the multiple-purpose nature of RWH. It is suggested that future work on RWH addresses three priority challenges. Firstly, more empirical data on system operation is needed to allow improved modelling by taking into account multiple objectives of RWH systems. Secondly, maintenance aspects and how they may impact the quality of collected rainwater should be explored in the future as a way to increase confidence on rainwater use. Finally, research should be devoted to the understanding of how institutional and socio-political support can be best targeted to improve system efficacy and community acceptance. [ABSTRACT FROM AUTHOR]

The aims of this study were to examine the removal of bacteria and viruses by household point-of-use (POU) treatments and to apply a previously developed large-volume virus concentration method (∼20 L). First, the removal of microbes by household POU treatment was investigated in the laboratory. Second, the prevalence of viruses in drinking water sources for households and the removal efficiency of microbes by POU treatments in two suburban communities in Hanoi, Vietnam, were investigated. Indigenous pepper mild mottle virus (PMMoV) was used as the main target together with adenovirus, Aichi virus, enterovirus, F-specific bacteriophage genogroup 1, andEscherichia colito investigate the removal efficiency of household treatments. The results from laboratory and field survey were compared. From the laboratory study, ceramic membranes were not effective for removing viruses and bacteria from water; pathogen reduction was less than 1.5 log10. By contrast, reverse osmosis (RO) devices reduced microbes by 3 to > 5 log10. In a field study, PMMoV was found to be the most prevalent waterborne virus. Household sand filtration was ineffective for removingE. coli, total coliforms and PMMoV; the reduction was less than 1 order of magnitude. Boiling the water and then filtering it with a ceramic membrane reducedE. coliby 3 orders of magnitude, but this was not effective for removing PMMoV. RO filtration was one of the promising methods for removingE. coli, total coliforms and PMMoV to below their detection limits in most of the samples studied. The removal ofE. coli, total coliforms and PMMoV was >2.3, >4 and >3 log10, respectively. The laboratory results of virus removal efficiency by POU devices agreed with the field study. Due to the prevalence and characteristics of PMMoV, it is a strong candidate for an indigenous indicator to investigate the viral removal efficiency of household POU treatments. [ABSTRACT FROM AUTHOR]

In order to properly assess and manage the risk of infection by enteric viruses in tap water, virus removal efficiency should be evaluated quantitatively for individual processes in actual drinking water treatment plants (DWTPs); however, there have been only a few studies due to technical difficulties in quantifying low virus concentration in water samples. In this study, the removal efficiency of indigenous viruses was evaluated for coagulation-sedimentation (CS) and rapid sand filtration (RSF) processes in a DWTP in Bangkok, Thailand by measuring the concentration of viruses before and after treatment processes using real-time polymerase chain reaction (qPCR). Water samples were collected and concentrated from raw source water, after CS, and after RSF, and inhibitory substances in water samples were reduced by use of a hydrophobic resin (DAX-8). Pepper mild mottle virus (PMMoV) and JC polyomavirus (JC PyV) were found to be highly prevalent in raw waters, with concentrations of 10 2.88 ± 0.35 and 10 3.06 ± 0.42 copies/L (geometric mean ± S.D.), respectively. Step-wise removal efficiencies were calculated for individual processes, with some variation observed between wet and dry seasons. During the wet season, PMMoV was removed less by CS and more by RSF on average (0.40 log 10 vs 1.26 log 10 , respectively), while the reverse was true for JC PyV (1.91 log 10 vs 0.49 log 10 , respectively). Both viruses were removed similarly during the dry season, with CS removing the most virus (PMMoV, 1.61 log 10 and 0.78 log 10 ; JC PyV, 1.70 log 10 , and 0.59 log 10 ; CS and RSF, respectively). These differences between seasons were potentially due to variations in raw water quality and the characteristics of the viruses themselves. These results suggest that PMMoV and JC PyV, which are more prevalent in environmental waters than the other enteric viruses evaluated in this study, could be useful in determining viral fate for the risk management of viruses in water treatment processes in actual full-scale DWTPs. [ABSTRACT FROM AUTHOR]

Reclaimed water has recently become an important water source for urban use, but the composition of dissolved organic matter (DOM) in reclaimed water has rarely been characterized at the compound level because of its complexity. In this study, the transformation and changes in composition of low molecular weight DOM in water reclamation processes, where secondary effluent of the municipal wastewater treatment plant was further treated by biofiltration, ozonation and chlorination, were investigated by “unknown” screening analysis using Orbitrap mass spectrometry (Orbitrap MS). The intense ions were detected over an m / z range from 100 to 450. In total, 2412 formulae with various heteroatoms were assigned, and formulae with carbon (C), hydrogen (H) and oxygen (O) only and C, H, O and sulfur (S) were the most abundant species. During biofiltration, CHO-only compounds with relatively high hydrogen to carbon (H/C) ratio or with saturated structure were preferentially removed, while CHOS compounds were mostly removed. Ozonation induced the greatest changes in DOM composition. CHOS compounds were mostly decreased after ozonation while ozone selectively removed CHO compounds with relatively unsaturated structure and produced compounds that were more saturated and with a higher degree of oxidation. After chlorination, 168 chlorine-containing formulae, chlorinated disinfection by-products (DBPs), were additionally detected. Candidate DBP precursors were determined by tracking chlorinated DBPs formed via electrophilic substitution, half of which were generated during the ozonation. [ABSTRACT FROM AUTHOR]

Ammonia is a precursor to trichloramine, which causes an undesirable chlorinous odor. Granular activated carbon (GAC) filtration is used to biologically oxidize ammonia during drinking water purification; however, little information is available regarding the abundance and diversity of ammonia-oxidizing archaea (AOA) and bacteria (AOB) associated with GAC. In addition, their sources and fates in water purification process remain unknown. In this study, six GAC samples were collected from five full-scale drinking water purification plants in Tokyo during summer and winter, and the abundance and community structure of AOA and AOB associated with GAC were studied in these two seasons. In summer, archaeal and bacterial amoA genes on GACs were present at 3.7 × 10-3.9 × 10 gene copies/g-dry and 4.5 × 10-4.2 × 10 gene copies/g-dry, respectively. In winter, archaeal amoA genes remained at the same level, while bacterial amoA genes decreased significantly for all GACs. No differences were observed in the community diversity of AOA and AOB from summer to winter. Phylogenetic analysis revealed high AOA diversity in group I.1a and group I.1b in raw water. Terminal-restriction fragment length polymorphism analysis of processed water samples revealed that AOA diversity decreased dramatically to only two OTUs in group I.1a after ozonation, which were identical to those detected on GAC. It suggests that ozonation plays an important role in determining AOA diversity on GAC. Further study on the cell-specific activity of AOA and AOB is necessary to understand their contributions to in situ nitrification performance. [ABSTRACT FROM AUTHOR]

The potential use of rainwater harvesting in conjunction with miscellaneous water supplies and a rooftop garden with rainwater harvesting facility for temperature reduction have been evaluated in this study for Hong Kong. Various water applications such as toilet flushing and areal climate controls have been systematically considered depending on the availability of seawater toilet flushing using the Geographic Information System (GIS). For water supplies, the district Area Precipitation per Demand Ratio (APDR) has been calculated to quantify the rainwater utilization potential of each administrative district in Hong Kong. Districts with freshwater toilet flushing prove to have higher potential for rainwater harvest and utilization compared to the areas with seawater toilet flushing. Furthermore, the effectiveness of using rainwater harvesting for miscellaneous water supplies in Hong Kong and Tokyo has been analyzed and compared; this revives serious consideration of diurnal and seasonal patterns of rainfall in applying such technology. In terms of the cooling effect, the implementation of a rooftop rainwater harvesting garden has been evaluated using the ENVI-met model. Our results show that a temperature drop of 1.3 °C has been observed due to the rainwater layer in the rain garden. This study provides valuable insight into the applicability of the rainwater harvesting for sustainable water management practice in a highly urbanized city. [ABSTRACT FROM AUTHOR]

Abstract: Rainfall events can introduce large amount of microbial contaminants including human enteric viruses into surface water by intermittent discharges from combined sewer overflows (CSOs). The present study aimed to investigate the effect of rainfall events on viral loads in surface waters impacted by CSO and the reliability of molecular methods for detection of enteric viruses. The reliability of virus detection in the samples was assessed by using process controls for virus concentration, nucleic acid extraction and reverse transcription (RT)-quantitative PCR (qPCR) steps, which allowed accurate estimation of virus detection efficiencies. Recovery efficiencies of poliovirus in river water samples collected during rainfall events (<10%) were lower than those during dry weather conditions (>10%). The log10-transformed virus concentration efficiency was negatively correlated with suspended solid concentration (r 2 =0.86) that increased significantly during rainfall events. Efficiencies of DNA extraction and qPCR steps determined with adenovirus type 5 and a primer sharing control, respectively, were lower in dry weather. However, no clear relationship was observed between organic water quality parameters and efficiencies of these two steps. Observed concentrations of indigenous enteric adenoviruses, GII-noroviruses, enteroviruses, and Aichi viruses increased during rainfall events even though the virus concentration efficiency was presumed to be lower than in dry weather. The present study highlights the importance of using appropriate process controls to evaluate accurately the concentration of water borne enteric viruses in natural waters impacted by wastewater discharge, stormwater, and CSOs. [Copyright &y& Elsevier]

Abstract: Nitrification is an important biological function of granular activated carbon (GAC) used in advanced drinking water purification processes. Newly discovered ammonia-oxidizing archaea (AOA) have challenged the traditional understanding of ammonia oxidation, which considered ammonia-oxidizing bacteria (AOB) as the sole ammonia-oxidizers. Previous studies demonstrated the predominance of AOA on GAC, but the contributions of AOA and AOB to ammonia oxidation remain unclear. In the present study, DNA-stable isotope probing (DNA-SIP) was used to investigate the autotrophic growth of AOA and AOB associated with GAC at two different ammonium concentrations (0.14 mg N/L and 1.4 mg N/L). GAC samples collected from three full-scale drinking water purification plants in Tokyo, Japan, had different abundance of AOA and AOB. These samples were fed continuously with ammonium and 13C-bicarbonate for 14 days. The DNA-SIP analysis demonstrated that only AOA assimilated 13C-bicarbonate at low ammonium concentration, whereas AOA and AOB exhibited autotrophic growth at high ammonium concentration. This indicates that a lower ammonium concentration is preferable for AOA growth. Since AOA could not grow without ammonium, their autotrophic growth was coupled with ammonia oxidation. Overall, our results point towards an important role of AOA in nitrification in GAC filters treating low concentration of ammonium. [Copyright &y& Elsevier]

Microbial regrowth causes problems during water reuse. Comprehensive understanding of the microorganisms that can regrow in reclaimed water and their substrate requirements are necessary. In this study, potential regrowth organisms were isolated from seven water reclamation plants in Japan. Based on 16S rDNA analysis, the isolates were grouped into 34 operational taxonomic units, belonging to Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes. Substrate utilization profiling using Biolog microplate™ classified the isolates into four groups. Bacteria in Cluster 1 (e.g., Methylobacterium sp. and Acinetobacter sp.) mainly utilized polymers, esters, amides, and alcohol. Isolates in Cluster 2 (e.g., Flavobacterium sp. and Microbacterium sp.) preferred to utilize polymers, carbohydrates, and esters. Isolates in Cluster 3 (e.g., Pseudomonas sp. and Acidovorax sp.) mainly utilized esters, carboxylic acids, and amino acids. Isolates in Cluster 4 (e.g., Enterobacter sp. and Rhodococcus sp.) utilized carbohydrates, esters, and amino acids. All isolates grew in reclaimed water treated by sand filtration, whereas some isolates could not grow in reclaimed water treated by coagulation and ozonation. Most bacteria in the same Biolog clusters exhibited similar growth characteristics in water samples. The potential of bacteria to regrow in reclaimed water likely depended on substrate requirement. [ABSTRACT FROM AUTHOR]

The goal of this study was to develop and validate a novel fosmid-clone-based metagenome isotope array approach - termed the community isotope array ( CIArray) - for sensitive detection and identification of microorganisms assimilating a radiolabeled substrate within complex microbial communities. More specifically, a sample-specific CIArray was used to identify anoxic phenol-degrading microorganisms in activated sludge treating synthetic coke-oven wastewater in a single-sludge predenitrification-nitrification process. Hybridization of the CIArray with DNA from the 14C-phenol-amended sample indicated that bacteria assimilating 14C-atoms, presumably directly from phenol, under nitrate-reducing conditions were abundant in the reactor, and taxonomic assignment of the fosmid clone end sequences suggested that they belonged to the Gammaproteobacteria. The specificity of the CIArray was validated by quantification of fosmid-clone-specific DNA in density-resolved DNA fractions from samples incubated with 13C-phenol, which verified that all CIArray-positive probes stemmed from microorganisms that assimilated isotopically labeled carbon. This also demonstrated that the CIArray was more sensitive than DNA- SIP, as the former enabled positive detection at a phenol concentration that failed to yield a 'heavy' DNA fraction. Finally, two operational taxonomic units distantly related to marine Gammaproteobacteria were identified to account for more than half of 16S r RNA gene clones in the 'heavy' DNA library, corroborating the CIArray-based identification. [ABSTRACT FROM AUTHOR]

Abstract: Microbial regrowth introduces several problems to the use of reclaimed water, such as health concerns, aesthetic deterioration, and biofouling. This study evaluated assimilable organic carbon (AOC), which is a part of the biodegradable organic matter promoting microbial growth, in water reclamation systems in Japan. The AOC concentration in the reclaimed water from various treatment processes ranged between 36 and 446 μg C/L (median 316 μg C/L). The AOC concentration in the reclaimed water from the plants equipped with ozonation was significantly higher – more than two times – than that in the reclaimed water from plants equipped with other processes. UV and chlorine also changed the AOC concentration slightly. Moreover, reverse osmosis produced reclaimed water with the lowest AOC content. Processes equipped with membrane filtration were effective in removing seed microorganisms that enter the distribution system. Microbial growth in reclaimed water distribution systems occurred when the total and free residual chlorine was lower than 0.36 and 0.09 mg/L, respectively. The AOC reduction occurred simultaneously with regrowth, which suggests that AOC could support microbial growth in reclaimed-water-distribution systems. As the residual chlorine is often depleted during distribution and storage, it is essential to control the AOC to suppress microbial growth. [Copyright &y& Elsevier]

The use of randomly generated DNA fragment sequences as probes on DNA arrays offers a unique potential for exploring unsequenced microorganisms. In this study, the detection specificity was evaluated with respect to probe-target sequence similarity using genomic DNAs of four Pseudomonas strains. Genome fragments averaging 2000 bp were found to be specific enough to discriminate 85-90% similarity under highly stringent hybridization conditions. Such stringent conditions compromised signal intensities; however, specific signals remained detectable at the highest stringency (at 75 °C hybridization) with negligible false negatives. These results suggest that, without any probe design or selection, genomic fragments can provide a reasonable specificity for microbial diagnostics or species delineation by DNA- DNA similarities. [ABSTRACT FROM AUTHOR]

Abstract: Microbial degradation of benzene under anaerobic conditions plays an important role in remediation of contaminated sites but the microorganisms and metabolic pathways involved remain poorly understood. In this study, we evaluated degradation of benzene by a methanogenic enrichment culture obtained from non-contaminated lotus field soil, alone and in the presence of several putative metabolic intermediates, that is, toluene, benzoate and phenol. Using stable isotope (13C substrate, benzene was shown to be degraded almost completely to equimolar concentrations of methane and carbon dioxide, without detectable accumulation of extracellular metabolites. Concurrently, toluene, benzoate and phenol were also effectively mineralized, but probably by microorganisms other than the benzene degraders. The latter included Hasda-A, which is putative benzene-degrading deltaproteobacterium present in the culture. While toluene and benzoate did not affect benzene degradation, phenol had a moderate inhibitory effect although it was not a major metabolic intermediate of benzene in our culture. Finally, 4-hydroxycoumarin was detected as a compound formed from phenol but further experiments are required to elucidate its relationship to degradation of phenol. [Copyright &y& Elsevier]

Abstract: Stable nitrogen and oxygen isotope ratios of nitrate (δ15N–NO3 and δ18O–NO3) have recently been used to identify nitrogen sources in water environments. However, there have been no investigations designed to determine nitrate isotopes in non-point sources in urban areas for evaluating the impact of surface deposits on nitrogen in surface runoff. In this study, we collected rainwater, surface runoff and surface deposits (road dust, roof dust and soil) to evaluate the nitrogen sources in surface runoff using nitrate isotopes. There were no large differences in δ15N–NO3 among rainwater (−0.3‰ to 1.5‰), surface runoff (−2.7‰ to 0.4‰), leachates from road dust (−5.8‰ to 6.2‰) and soil (−11.5‰ to 0.6‰). In contrast, the δ18O–NO3 in surface runoff (28.5–47.9‰) was lower than that in rainwater (62.7–78.6‰), and higher than that in leachates from road dust (6.1–27.6‰) and soil (−1.1‰ to 6.6‰). δ18O–NO3 is a useful indicator for evaluating the NO3–N sources in surface runoff. Using this indicator, NO3–N from road dust was estimated to account for more than half of the NO3–N in surface runoff. This is consistent with a result based on a comparison of their loads per unit surface between rainwater and surface runoff, which also showed that most of the nitrogen in surface runoff was derived from surface deposits. [Copyright &y& Elsevier]

Inhibitors that reduce viral nucleic acid extraction efficiency and interfere with cDNA synthesis and/or polymerase activity affect the molecular detection of viruses in aquatic environments. To overcome these significant problems, we developed a methodology for assessing nucleic acid yields and DNA amplification efficiencies for environmental water samples. This involved adding particles of adenovirus type 5 and murine norovirus and newly developed primer-sharing controls, which are amplified with the same primer pairs and result in the same amplicon sizes as the targets, to these samples. We found that nucleic acid loss during the extraction process, rather than reverse transcription-PCR (RT-PCR) inhibition, more significantly attributed to underestimation of the presence of viral genomes in the environmental water samples tested in this study. Our success rate for satisfactorily amplifying viral RNAs and DNAs by RT-PCR was higher than that for obtaining adequate nucleic acid preparations. We found that inhibitory properties were greatest when we used larger sample volumes. A magnetic silica bead-based RNA extraction method effectively removed inhibitors that interfere with viral nucleic acid extraction and RT-PCR. To our knowledge, this is the first study to assess the inhibitory properties of environmental water samples by using both control virus particles and primer-sharing controls. [ABSTRACT FROM AUTHOR]

Abstract: Road dust is considered an important source of sediment contamination; however, there are few studies on the toxicity of road dust on benthic organisms. This study evaluates the toxicity of road dust on the benthic ostracod, Heterocypris incongruens, through a 6-day direct exposure experiment. All six road dust samples collected in heavy traffic areas caused high mortality of the ostracod, whereas the road dust collected in a residential area did not show toxicity to the ostracod. After maintaining the mixture of road dust and water for 24h, the toxicity of the road dust decreased significantly for three of the six samples in the heavy traffic areas, suggesting these road dust samples become less toxic in the surface runoff process. In addition, we conducted the same toxicity test on manipulated road dust using different solid/liquid ratios and holding times to evaluate the change in toxicity caused by the runoff process. Wet road dust that had been separated from a dust–water mixture after a holding time of 1h or 24h did not show lethal toxicity, while the water-soluble fraction of the mixture caused high mortality of the ostracod at a solid/liquid ratio of 1:2 and 1:4. However, after a 7-day holding time of the dust–water mixture, the wet road dust and the water-soluble fraction showed lethal toxicity to the ostracod. These results suggest that toxicants of road dust seemed to exist mainly in water soluble fraction eluted off by rain water; however, particle-bound fraction again showed lethal toxicity after 7days of incubation. [Copyright &y& Elsevier]

Abstract: Ozonation followed by granular activated carbon (GAC) is one of the advanced drinking water treatments. During GAC treatment, ammonia can be oxidized by ammonia-oxidizing microorganisms associated with GAC. However, there is little information on the abundance and diversity of ammonia-oxidizing microorganisms on GAC. In this study, the nitrification activity of GAC and the settlement of ammonia-oxidizing archaea (AOA) and bacteria (AOB) in GAC were monitored at a new full-scale advanced drinking water treatment plant in Japan for 1 year after plant start-up. Prechlorination was implemented at the receiving well for the first 10 months of operation to treat ammonia in raw water. During this prechlorination period, levels of both AOA and AOB associated with GAC were below the quantification limit. After prechlorination was stopped, 105 copies g-dry−1 of AOA amoA genes were detected within 3 weeks and the quantities ultimately reached 106–107 copies g-dry−1, while levels of AOB amoA genes still remained below the quantification limit. This observation indicates that AOA can settle in GAC rapidly without prechlorination. The nitrification activity of GAC increased concurrently with the settlement of AOA after prechlorination was stopped. Estimation of in situ cell-specific ammonia-oxidation activity for AOA on the assumption that only AOA and AOB determined can contribute to nitrification suggests that AOA may account for most of the ammonia-oxidation. However, further validation on AOB contribution is required. [ABSTRACT FROM AUTHOR]

Abstract: Inefficient removal of estrone (E1) in wastewater treatment plants (WWTPs) causes feminizing effects in male aquatic creatures. As E1 is mainly removed by biodegradation, investigation of E1 degradation is important to determine better removal strategies. Using microautoradiography-fluorescence in situ hybridization (MAR-FISH), we demonstrated that the structures of [3H]E1-incorporating bacterial communities were different at different E1 concentrations applied to activated sludge. At 200 μg/L E1, almost all [3H]E1-incorporating cells were associated with either Betaproteobacteria or Gammaproteobacteria (60% and 40% of MAR (+) cells, respectively). The proportion of Betaproteobacteria and Gammaproteobacteria in the total number of [3H]E1-incorporating cells decreased as the concentration of E1 decreased. In contrast, the proportion of Alphaproteobacteria in the total number of [3H]E1-incorporating cells increased as the concentrations of E1 decreased. At the lowest applied concentration (540 ng/L), almost all the [3H]E1-incorporating cells were Alphaproteobacteria (96%). The results of MAR-FISH applied to sludge samples collected from various plant locations and activated sludge processes, and during different seasons also demonstrated the high contribution of Alphaproteobacteria to the entire E1-degrading bacterial community (50.4 ± 11% of the total number of [3H]E1-incorporating cells) at 1 μg/L E1. Since the E1 concentration in domestic wastewater is at sub-μg/L levels, the key E1 degraders in activated sludge of domestic WWTPs are probably be Alphaproteobacteria. All [3H]E1-incorporating Alphaproteobacteria were hybridized with probe ALF968. Few MAR (+) cells were Sphingomonadales. An E1-degrading bacterial community at low E1 concentration appeared to consist of diverse bacterial groups of Alphaproteobacteria. This study suggested that substrate concentration is an essential factor for revealing E1-degrading bacteria in complex communities. [Copyright &y& Elsevier]

The effects of different placements of rapid sand filtration on nitrification performance of BAC treatment in a pilot-scale plant were evaluated. In this plant, rapid sand filtration was placed after ozonation-BAC treatment in Process (A), while it preceded ozonation-BAC treatment in Process (B). Analysis of amoA genes of ammonia-oxidizing archaea (AOA) and bacteria (AOB) combined with nitrification potential test was conducted. BAC from Process (A) demonstrated slightly higher nitrification potential at every sampling occasion. This might be due to higher abundances of AOB on BAC from Process (A) than those on BAC from Process (B). However, AOA rather than AOB could be predominant ammonia-oxidizers in BAC treatment regardless of the position of rapid sand filtration. The highest nitrification potential was observed for BAC from both processes in February when the highest abundances of AOA-amoA and AOB-amoA genes were detected. Since rapid sand filtration was placed after BAC treatment in Process (A), residual aluminum concentration in BAC influent was higher in Process (A). However, adverse effects of aluminum on nitrification activity were not observed. These results suggest that factors other than aluminum concentration in different treatment processes could possibly have some influence on abundances of ammonia-oxidizing microorganisms on BAC. [ABSTRACT FROM AUTHOR]

In Japan, water supply utilities need to renovate their sludge treatment facilities. This study therefore evaluated treatment of the sludge from drinking water treatment plants (DWTP) by two approaches: an individual system and a combined system. In the individual treatment scenario, the DWTP treats the sludge using its own facilities, whereas in the combined treatment scenario, DWTP sludge is transported to a sewerage system. The construction cost is much lower for combined treatment than individual treatment, while the operation cost is higher for combined treatment owing to the sewer charge. However, the additional sewer charge is less than the increase in cost for constructing an individual sludge treatment system. With the combined treatment system, sludge must be transported from the DWTP to a sewage sludge treatment plant (SSTP). A cost analysis reveals that sludge transportation by road is less beneficial than by pipeline in the sewerage system unless the plants are nearby. In economic terms, combined treatment and a sludge liquor pipeline is the most advantageous of all cases. The total cost depends on the given conditions and assumed unit costs. Sensitive parameters are the sewer charge and the distance between the DWTP and sewer pipeline. [ABSTRACT FROM AUTHOR]

Diverse environmental genes have been identified recently. To characterize their functions, it is necessary to understand which genes and what combinations of those genes are responsible for the biodegradation of soil contaminants. In this article, a 60-mer oligonucleotide microarray was constructed to simultaneously detect di- and monooxygenase genes for benzene and related compounds. In total, 148 probes were designed and validated by pure-culture hybridizations using the following criteria to discriminate between highly homologous genes: ≤53-bp identities and ≤25-bp continuous stretch to nontarget sequences. Microarray hybridizations were performed using PCR products amplified from five benzene-amended soils and two oil-contaminated soils. Six of the probes gave a positive signal for more than six soils; thus, they may represent key sequences for benzene degradation in the environment. The microarray developed in this study will be a powerful tool for the screening of key genes involved in benzene degradation and for the rapid profiling of benzene oxygenase gene diversity in contaminated soils. [ABSTRACT FROM AUTHOR]