Autotrophic growth activity of complete ammonia oxidizers in an upflow biological contact filter for drinking water treatment.

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.

Identification of new eligible indicator organisms for combined sewer overflow via 16S rRNA gene amplicon sequencing in Kanda River, Tokyo.

Fecal indicator bacteria (FIB) are commonly used to evaluate the pollution impact of combined sewer overflows (CSOs) in urban rivers. Although water quality assessment with FIB has a long tradition, recent studies demonstrated that FIB have a low correlation with pathogens and therefore are not accurate enough for the assessment of potential human hazards in water. Consequently, new eligible and more specific indicators have to be identified, which was done in this study via sequencing of genetic markers from total community DNA. To identify potential microbiome-based indicators, microbial communities in samples from an urban river in Tokyo under different climatic conditions (dry and rainy) were compared with the influent and effluent of three domestic wastewater treatment plants (WWTPs) by analyzing 16 S rRNA gene amplicon libraries. In the first part of this study, physicochemical parameters and FIB quantification with selective culture techniques facilitated the identification of samples contaminated with CSO, sewage, or both. This allowed the grouping of samples into CSO-contaminated and non-contaminated samples, an essential step prior to the microbiome comparison between samples. Increased turbidity, ammonia concentrations, and E. coli [up to (9.37 ± 0.95) × 102 CFU/mL after 11.5 mm of rainfall] were observed in CSO-contaminated river samples. Comparison of dry weather (including WWTP samples) and rainy weather samples showed a reduction in microbial diversity in CSO-contaminated samples. Furthermore, the results of this study suggest Bacteroides spp. as a novel indicator of sewage pollution in surface waters.

Temperature-Dependent Ammonium Removal Capacity of Biological Activated Carbon Used in a Full-Scale Drinking Water Treatment Plant.

Nitrification is a key function of biological activated carbon (BAC) filters for drinking water treatment. It is empirically known that the nitrification activity of BAC filters depends on water temperature, potentially resulting in the leakage of ammonium from BAC filters when the water temperature decreases. However, the ammonium removal capacity of BAC filters and factors governing the capacity remain unknown. This study employed a bench-scale column assay to determine the volumetric ammonium removal rate (VARR) of BAC collected from a full-scale drinking water treatment plant. VARR was determined at a fixed loading rate under different conditions. Seasonal variations of the VARR as well as impacts of the water matrix and water temperature on ammonium removal were quantitatively analyzed. While the VARR in an inorganic medium at 25 °C was maintained even during low water temperature periods and during breakpoint chlorination periods, the water matrix factor reduced the VARR in ozonated water at 25 °C by 33% on average. The VARR in ozonated water was dependent on water temperature, indicating that the microbial activity of BAC did not adapt to low water temperature. The Arrhenius equation was applied to reveal the relationship between VARR and water temperature. The actual ammonium removal performance of a full-scale BAC filter was predicted. VARR is useful for water engineers to reexamine the loading and filter depth of BAC filters.

Changes in Dissolved Organic Matter Composition and Disinfection Byproduct Precursors in Advanced Drinking Water Treatment Processes.

Molecular changes in dissolved organic matter (DOM) from treatment processes at two drinking water treatment plants in Japan were investigated using unknown screening analysis by Orbitrap mass spectrometry. DOM formulas with carbon, hydrogen and oxygen (CHO-DOM) were the most abundant class in water samples, and over half of them were commonly found at both plants. Among the treatment processes, ozonation induced the most drastic changes to DOM. Mass peak intensities of less saturated CHO-DOM (positive (oxygen subtracted double bond equivalent per carbon (DBE-O)/C)) decreased by ozonation, while more saturated oxidation byproducts (negative (DBE-O)/C) increased and new oxidation byproducts (OBPs) were detected. By Kendrick mass analysis, ozone reactions preferred less saturated CHO-DOM in the same alkylation families and produced more saturated alkylation families of OBPs. Following ozonation, biological activated carbon filtration effectively removed <300 Da CHO-DOM, including OBPs. Following chlorination, over 50 chlorinated formulas of disinfection byproducts (DBPs) were found in chlorinated water samples where at least half were unknown. Putative precursors of these DBPs were determined based on electrophilic substitutions and addition reactions. Ozonation demonstrated better decomposition of addition reaction-type precursors than electrophilic substitution-type precursors; over half of both precursor types decreased during biological activated carbon filtration.

Carcinogenic profile, toxicity and source apportionment of polycyclic aromatic hydrocarbons accumulated from urban road dust in Tokyo, Japan.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in urban environments. Urban road dust (URD) generated by traffic is an important PAH accumulator. Twelve priority PAHs in < 2000 µm fraction of ten URD samples from Tokyo, Japan were characterized based on profile distributions, carcinogenicity, toxicity, and source apportionment by cluster analysis, biplot and diagnostic ratios. PAH concentrations (mg/kg dry weight) in arterial roads, highways, highway parking, highway drainage pit and residential area URD samples were 2.06-4.24, 0.25-3.37, 3.44, 4.94, and 5.26 respectively, dominated by the ∑4 rings (average 46%) and ∑5 + 6 rings (average 41%) PAHs. Biplot analysis revealed that the antecedent dry weather period, vehicle frequency and organic matter content were the dominant environmental factors governing PAH profiles of different road types. The total amount of carcinogenic PAHs in the residential URD (2.12 mg/kg) was higher than those in the arterial road (0.60-2.00 mg/kg) and highway (0.10-1.84 mg/kg) URD. Toxic equivalent concentrations (TECs) of residential, arterial road and highway URD were 0.54, (0.12-0.57), and (0.02-0.51) mg/kg, respectively. The dominant PAH sources were found to be petrogenic combustion in arterial road and highway URD, and pyrogenic combustion consisting of a mix of biomass, petroleum and traffic-related sources in the residential and highway drainage pit samples. This is also the first study to find that TEC-based toxicity should not be taken as a measure of URD toxicity.

Characterization of Natural Organic Substances Potentially Hindering RT-PCR-Based Virus Detection in Large Volumes of Environmental Water.

Quantitative detection of pathogenic viruses in the environmental water is essential for the assessment of water safety. It is known that some of natural organic substances interfere with virus detection processes, i.e., nucleic acid extraction and reverse transcription-PCR. Such substances are carried over into a sample after virus concentration. In this study, inhibitory substances in coastal water samples were characterized in view of their effects on efficiency of virus detection and property as organic matters. Among 81 samples tested, 77 (95%) showed low recoveries (<10%) of spiked murine norovirus. These recovery rates were correlated with the levels of organic matter present in virus concentrates as measured by ultraviolet absorbance at 254 nm (r = -0.70 - -0.71, p < 0.01). High-performance gel chromatography and fluorescence excitation-emission matrix spectroscopy revealed that organic fractions in the 10-100 kDa size range, which were not dominant in the original samples, and those possessing humic acid-like fluorescence properties were dominant in virus concentrates. The inhibitory effect was more pronounced during summer. Substances originating from seawater seemed to cause a more pronounced effect than those originating from wastewater. Our data highlight the previously unknown characteristics of natural inhibitory substances and are helpful in establishing an effective sample purification technique.

Abundance and diversity of ammonia-oxidizing archaea and bacteria on granular activated carbon and their fates during drinking water purification process.

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(5)-3.9 × 10(8) gene copies/g-dry and 4.5 × 10(6)-4.2 × 10(8) 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.

Characterization of Washoff Behavior of In-Sewer Deposits in Combined Sewer Systems.

In-sewer deposits in combined sewer systems (CSSs) are closely related with the behavior of first foul flush and combined sewer overflows. The artificial flushing experiment separating the washoff of in-sewer deposits from the inflow of surface pollutants was carried out to simulate first foul flush in a CSS. The washoff behaviors of each pollutant including chemical pollutants, bacterial indicators, and enteric viruses were intensively investigated. By using several morphological analyses, some of which were suggested through this study, the characteristics of first foul flush were examined. As a result, the washoff behaviors of each pollutant were characterized according to their (i) event load ratios (ELRs), (ii) time-series concentration and load curves, (iii) concentration vs. flow rate curves, and (iv) dimensionless runoff concentrations (DRCs). The first foul flush patterns of each parameter were categorized into 3 typical groups: the strong-, partial-, and no first foul flush group. The order of these groups signifies their different physicochemical properties of in-sewer deposits in CSSs, their strength of first foul flush phenomena, and the washoff priority as well.

Impacts of urbanization on the prevalence of antibiotic-resistant Escherichia coli in the Chaophraya River and its tributaries.

River water samples were taken from 32 locations around the basin of Chaophraya River and its four major tributaries in Thailand to investigate resistance ratios of Escherichia coli isolates to eight antibiotic agents of amoxicillin, sulfamethoxazole/trimethoprim, tetracycline, doxytetracycline, ciprofloxacin, levofloxacin, norfloxacin and ofloxacin. Principal component analysis was performed to characterize resistance patterns of the samples. Relevancy of the obtained principal components with urban land use and fecal contamination of the river were examined. The ratio of antibiotic-resistant bacteria is likely to increase when urban land use near the sampling site exceeds a certain ratio. The resistance ratio to fluoroquinolones tends to be high in a highly populated area. Meanwhile, no significant contribution of fecal contamination was found to increase the resistance ratio. These results suggest that an antibiotic-resistance ratio is dependent on conditions of local urbanization rather than the upstream conditions, and that the major sources of antibiotic-resistant bacteria in the Chaophraya River basin are possibly point sources located in the urban area which contains a high ratio of resistant bacteria.

Detection of pepper mild mottle virus as an indicator for drinking water quality in Hanoi, Vietnam, in large volume of water after household treatment.

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, and Escherichia coli to 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 removing E. 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 reduced E. coli by 3 orders of magnitude, but this was not effective for removing PMMoV. RO filtration was one of the promising methods for removing E. coli, total coliforms and PMMoV to below their detection limits in most of the samples studied. The removal of E. 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.

Organic substances interfere with reverse transcription-quantitative PCR-based virus detection in water samples.

Reverse transcription (RT)-PCR-based virus detection from water samples is occasionally hampered by organic substances that are co-concentrated during virus concentration procedures. To characterize these organic substances, samples containing commercially available humic acid, which is known to inhibit RT-PCR, and river water samples were subjected to adsorption-elution-based virus concentration using an electronegative membrane. In this study, the samples before, during, and after the concentration were analyzed in terms of organic properties and virus detection efficiencies. Two out of the three humic acid solutions resulted in RT-quantitative PCR (qPCR) inhibition that caused >3-log10-unit underestimation of spiked poliovirus. Over 60% of the organics contained in the two solutions were recovered in the concentrate, while over 60% of the organics in the uninhibited solution were lost during the concentration process. River water concentrates also caused inhibition of RT-qPCR. Organic concentrations in the river water samples increased by 2.3 to 3.9 times after the virus concentration procedure. The inhibitory samples contained organic fractions in the 10- to 100-kDa size range, which are suspected to be RT-PCR inhibitors. According to excitation-emission matrices, humic acid-like and protein-like fractions were also recovered from river water concentrates, but these fractions did not seem to affect virus detection. Our findings reveal that detailed organic analyses are effective in characterizing inhibitory substances.

Wastewater Analysis Indicates that Genetically Diverse Astroviruses, Including Strains Belonging to Novel Clades MLB and VA, Are Circulating within Japanese Populations.

Human astroviruses (HAstVs) are a common etiological agent of infantile gastroenteritis. Recent studies revealed that novel astrovirus (AstV) strains of the MLB clade (MLB-AstVs) and VA clade (VA-AstVs), which are genetically distinct from the classic HAstVs, are circulating in the human population. In the present study, we quantified classic HAstVs as well as carried out a genetic analysis of classic and novel HAstVs in wastewater in Japan. The concentration of classic HAstVs in the influent water samples ranged from 10(4) to 10(5) copies per liter, and the amount removed by wastewater treatment was determined to be 2.4 ± 0.3 log10. Four types of classic HAstV strains (HAstV types 1, 2, 5, and 4/8) as well as novel AstV strains belonging to the MLB-2, VA-1, and VA-2 clades were identified using reverse transcription-PCR (RT-PCR) assays, including assays newly developed for the detection of strains of the MLB and VA clades, followed by cloning and nucleotide sequencing. Our results suggest that genetically diverse AstV strains are circulating among the human population in Japan. The newly developed (semi)nested RT-PCR assays for these novel AstV clades are useful to identify and characterize the novel AstVs in environmental waters.

Beneath the surface: Exploring microplastic intricacies in Anadara granosa.

Anadara granosa or blood cockles have been reported to be a candidate for biomonitoring agents due to their sedimentary nature and their nutrient uptake mechanisms. Yet, this bivalve is still regarded as a delicacy in Asian cuisine. Malaysia is the largest exporter of this sea product that contaminated cockles may also be experienced by the importing countries. However, the bioaccumulation of microplastics in A. granosa cultivated in Malaysia has not been extensively studied. It is crucial to comprehend the risk posed to humans by consuming A. granosa in their diet. Therefore, the purpose of this research is to investigate the levels of microplastic accumulation in A. granosa from major exporters in Peninsular Malaysia, to evaluate the associated risk of microplastics on the species, and to estimate daily human consumption of microplastics through the consumption of A. granosa. The abundance of microplastics was quantified through the use of a stereo microscope, and the polymer type was determined using FTIR and micro-FTIR. Findings from this investigation revealed that all samples of A. granosa were contaminated with microplastics, with the highest levels of accumulation found in bivalves collected from the west coast (0.26 ± 0.15 particles/g) of Peninsular Malaysia. Fragment and fiber microplastics, measuring between 0.05 and 0.1 mm in size, were found to be the most prevalent in A. granosa, with blue being the dominant identified colour and rayon being the most common polymer type. Microplastic risk assessment due to the presence of polyacrylate, polycarbonate (PC), and polymethyl methacrylate (PMMA) resulted in a high risk of contamination for A. granosa. It was further determined that the current estimated dietary intake (EDI) suggests that consumers of A. granosa uptake approximately 21.8-93.5 particles/person/year of microplastics. This study highlights that A. granosa accumulates microplastics, which could potentially result in bioaccumulation and biomagnification in humans through consumption.

Non-target liquid chromatography high-resolution mass spectrometry screening to prioritize unregulated micropollutants that persist through domestic wastewater treatment.

Efforts to regulate and monitor emerging contaminants are insufficient because new chemicals are continually brought to market, and many are unregulated and potentially harmful. Domestic wastewater treatment plants are not designed to remove micropollutants and are important sources of emerging contaminants in the aquatic environment. In this study, non-target screening, an unbiased method for analyzing compounds without prior information, was used to identify compounds that may be emitted in wastewater treatment plant effluent and should be monitored. Nine wastewater treatment plants using different treatment methods were studied, and a non-target screening data-processing method was used. The frequencies at which the contaminants were detected and contaminant persistence through the treatment processes were considered, and then the contaminants were prioritized. The predicted no-effect concentration of each prioritized contaminant was used to determine whether further analysis and monitoring of the contaminant was necessary. Quantitative analyses of five compounds (amantadine, atenolol, benzotriazole, diphenhydramine, and sulpiride) were performed using reference standards. Probable molecular formulae and structures were proposed for 17 contaminants, and the risks posed by the contaminants were estimated using predicted no-effect concentrations. The results provide valuable insights into how unregulated micropollutants can be identified and prioritized for monitoring in future studies.

Supporting data-enhanced hybrid ordinary differential equation model for phosphate dynamics in municipal wastewater treatment.

A parallel hybrid ordinary differential equation (ODE) integrating the Activated Sludge Model No. 2d (ASM2d) and an artificial neural network (ANN) was developed to simulate biological phosphorus removal (BPR) with high accuracy and interpretability. Two novelties were introduced; first, the involved supporting data (i.e., phosphate-release activity) were incorporated as an input in the ANN. Second, the outputs of the ANN were selective. Three models were implemented using different ANN outputs, and all three outperformed ASM2d in phosphate estimation for anaerobic/aerobic sequencing batch reactor operation. In particular, the incorporation of four variables responsible for BPR into the ANN enabled the highest performance (R2 = 0.93) owing to the capture of increasing phosphate-accumulating organisms (PAOs). The ANN with the supporting data worked satisfactorily to compensate for ASM2d by adding proper PAOs, resulting in improvement in phosphate estimation. The novel parallel hybrid ODE can simulate BPR while maintaining physical meaning.

Characterization of bacterial isolates from water reclamation systems on the basis of substrate utilization patterns and regrowth potential in reclaimed water.

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.

Cluster analysis for characterization of rainfalls and CSO behaviours in an urban drainage area of Tokyo.

Combined sewer overflow (CSO) from urban areas is recognized as a major pollutant source to the receiving waters during wet weather. This study attempts to categorize rainfall events and corresponding CSO behaviours to reveal the relationship between rainfall patterns and CSO behaviours in the Shingashi urban drainage areas of Tokyo, Japan where complete service by a combined sewer system (CSS) and CSO often takes place. In addition, outfalls based on their annual overflow behaviours were characterized for effective storm water management. All 117 rainfall events recorded in 2007 were simulated by a distributed model InfoWorks CS to obtain CSO behaviours. The rainfall events were classified based on two sets of parameters of rainfall pattern as well as CSO behaviours. Clustered rainfall and CSO groups were linked by similarity analysis. Results showed that both small and extreme rainfalls had strong correlations with the CSO behaviours, while moderate rainfall had a weak relationship. This indicates that important and negligible rainfalls from the viewpoint of CSO could be identified by rainfall patterns, while influences from the drainage area and network should be taken into account when estimating moderate rainfall-induced CSO. Additionally, outfalls were finally categorized into six groups indicating different levels of impact on the environment.

Variation in PAH patterns in road runoff.

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.

Monsoon dilutes the concurrence but increases the correlation of viruses and Pharmaceuticals and Personal Care Products (PPCPs) in the urban waters of Guwahati, India: The context of pandemic viruses.

Concurrence of pharmaceuticals and personal care products (PPCPs), pathogenic viruses, metals and microbial pollution along with their seasonal variations in the water environment are overarching in the context of existing pandemic, especially for tropical countries. The present study focuses on the seasonal influence on the vulnerability of urban water in Guwahati, the largest city in North-eastern India, through examining the concurrence of seven PPCPs, five viruses, faecal bacteria and nine metals in surface waters during monsoon (Summer-July 2017) and pre-monsoon (Winter-March 2018). Surface water sampling was carried out at different locations of the Brahmaputra River, its tributary Bharalu River (an unlined urban drain), and Dipor Bill Lake (Ramsar-recognized wetland). Both PPCPs and viruses were at high concentrations (e.g. up to 970 ng L-1 caffeine, 2.5 × 103 copies mL-1 pepper mild mottle virus (PMMoV)) at the confluence points of urban drains and the river, while they were mostly undetectable at both upstream and downstream locations, implying strong self-purification ability of the river. All the analysed PPCPs and viruses were at much higher concentrations during pre-monsoon i.e., winter than during monsoon, implying heavy dilution and temperature effect during the monsoon. Overall, PPCPs and viruses were more correlated in monsoon but the risk quotient in the urban tributary was higher in pre-monsoon (e.g. 5061 in pre-monsoon and 1515 in monsoon for caffeine). PMMoV was found to be an excellent faecal pollution indicator due to its prevalence, detectability and specificity in all seasons. Overall, the seasonal fluctuations of the non-enveloped viruses monitored in this study is likely to be relevant for SARS-CoV-2. We contribute to address the literature scarcity pertaining to seasonal variations in the prevalence of viruses and their concurrences with contaminants of emerging concern.

Dynamics of the Microbial Community and Opportunistic Pathogens after Water Stagnation in the Premise Plumbing of a Building.

In premise plumbing, microbial water quality may deteriorate under certain conditions, such as stagnation. Stagnation results in a loss of disinfectant residual, which may lead to the regrowth of microorganisms, including opportunistic pathogens. In the present study, microbial regrowth was investigated at eight faucets in a building over four seasons in one year. Water samples were obtained before and after 24 h of stagnation. In the first 100| |mL after stagnation, total cell counts measured by flow cytometry increased 14- to 220-fold with a simultaneous decrease in free chlorine from 0.17-0.36| |mg L-1 to <0.02| |mg L-1. After stagnation, total cell counts were not significantly different among seasons; however, the composition of the microbial community varied seasonally. The relative abundance of Pseudomonas spp. was dominant in winter, whereas Sphingomonas spp. were dominant in most faucets after stagnation in other seasons. Opportunistic pathogens, such as Legionella pneumophila, Mycobacterium avium, Pseudomonas aeruginosa, and Acanthamoeba spp., were below the quantification limit for real-time quantitative PCR in all samples. However, sequences related to other opportunistic pathogens, including L. feeleii, L. maceachernii, L. micdadei, M. paragordonae, M. gordonae, and M. haemophilum, were detected. These results indicate that health risks may increase after stagnation due to the regrowth of opportunistic pathogens.