Effect-based monitoring to integrate the mixture hazards of chemicals into water safety plans.

Water safety plans (WSPs) are intended to assure safe drinking water (DW). WSPs involve assessing and managing risks associated with microbial, chemical, physical and radiological hazards from the catchment to the consumer. Currently, chemical hazards in WSPs are assessed by targeted chemical analysis, but this approach fails to account for the mixture effects of the many chemicals potentially present in water supplies and omits the possible effects of non-targeted chemicals. Consequently, effect-based monitoring (EBM) using in vitro bioassays and well plate-based in vivo assays are proposed as a complementary tool to targeted chemical analysis to support risk analysis, risk management and water quality verification within the WSP framework. EBM is frequently applied to DW and surface water and can be utilised in all defined monitoring categories within the WSP framework (including 'system assessment', 'validation', 'operational' and 'verification'). Examples of how EBM can be applied within the different WSP modules are provided, along with guidance on where to apply EBM and how frequently. Since this is a new area, guidance documents, standard operating procedures (SOPs) and decision-making frameworks are required for both bioassay operators and WSP teams to facilitate the integration of EBM into WSPs, with these resources being developed currently.

Monitoring of SARS-CoV-2 in wastewater: what normalisation for improved understanding of epidemic trends?

SARS-CoV-2 RNA quantification in wastewater has emerged as a relevant additional means to monitor the COVID-19 pandemic. However, the concentration can be affected by black water dilution factors or movements of the sewer shed population, leading to misinterpretation of measurement results. The aim of this study was to evaluate the performance of different indicators to accurately interpret SARS-CoV-2 in wastewater. Weekly/bi-weekly measurements from three cities in France were analysed from February to September 2021. The concentrations of SARS-CoV-2 gene copies were normalised to the faecal-contributing population using simple sewage component indicators. To reduce the measurement error, a composite index was created to combine simultaneously the information carried by the simple indicators. The results showed that the regularity (mean absolute difference between observation and the smoothed curve) of the simple indicators substantially varied across sampling points. The composite index consistently showed better regularity compared to the other indicators and was associated to the lowest variation in correlation coefficient across sampling points. These findings suggest the recommendation for the use of a composite index in wastewater-based epidemiology to compensate for variability in measurement results.

Elimination of SARS-CoV-2 along wastewater and sludge treatment processes.

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is shed in the feces of infected people. As a consequence, genomic RNA of the virus can be detected in wastewater. Although the presence of viral RNA does not inform on the infectivity of the virus, this presence of genetic material raised the question of the effectiveness of treatment processes in reducing the virus in wastewater and sludge. In this work, treatment lines of 16 wastewater treatment plants were monitored to evaluate the removal of SARS-CoV-2 RNA in raw, processed waters and sludge, from March to May 2020. Viral RNA copies were enumerated using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in 5 different laboratories. These laboratories participated in proficiency testing scheme and their results demonstrated the reliability and comparability of the results obtained for each one. SARS-CoV-2 RNA was found in 50.5% of the 101 influent wastewater samples characterized. Positive results were detected more frequently in those regions with a COVID-19 incidence higher than 100 cases per 100,000 inhabitants. Wastewater treatment plants (WWTPs) significantly reduced the occurrence of virus RNA along the water treatment lines. Secondary treatment effluents showed an occurrence of SARS-CoV-2 RNA in 23.3% of the samples and no positive results were found after MBR and chlorination. Non-treated sludge (from primary and secondary treatments) presented a higher occurrence of SARS-CoV-2 RNA than the corresponding water samples, demonstrating the affinity of virus particles for solids. Furthermore, SARS-CoV-2 RNA was detected in treated sludge after thickening and anaerobic digestion, whereas viral RNA was completely eliminated from sludge only when thermal hydrolysis was applied. Finally, co-analysis of SARS-CoV-2 and F-specific RNA bacteriophages was done in the same water and sludge samples in order to investigate the potential use of these bacteriophages as indicators of SARS-CoV-2 fate and reduction along the wastewater treatment.

Faster and safer: Research priorities in water and health.

The United Nations' Sustainable Development Goals initiated in 2016 reiterated the need for safe water and healthy lives across the globe. The tenth anniversary meeting of the International Water and Health Seminar in 2018 brought together experts, students, and practitioners, setting the stage for development of an inclusive and evidence-based research agenda on water and health. Data collection relied on a nominal group technique gathering perceived research priorities as well as underlying drivers and adaptation needs. Under a common driver of public health protection, primary research priorities included the socioeconomy of water, risk assessment and management, and improved monitoring methods and intelligence. Adaptations stemming from these drivers included translating existing knowledge to providing safe and timely services to support the diversity of human water needs. Our findings present a comprehensive agenda of topics at the forefront of water and health research. This information can frame and inform collective efforts of water and health researchers over the coming decades, contributing to improved water services, public health, and socioeconomic outcomes.

Non-tuberculous mycobacteria in drinking water systems: A review of prevalence data and control means.

Non-tuberculous species of Mycobacterium are commonly found in a large diversity of water environments, and epidemiological studies suggest that natural or drinking waters are the principal sources of human contamination. Controlling non-tuberculous mycobacteria in water systems is therefore important to prevent infection with these micro-organisms. This review article summarizes the information and data published up to now on the factors favoring the presence of these bacteria in natural and artificial water systems, the effectiveness of water treatment means, and based on this information, identifies possible means to control the presence of non-tuberculous mycobacteria in drinking water.

Reduced Chlorine in Drinking Water Distribution Systems Impacts Bacterial Biodiversity in Biofilms.

In drinking water distribution systems (DWDS), a disinfectant residual is usually applied to limit bacterial regrowth. However, delivering water with no or reduced chlorine residual could potentially decrease the selection for antimicrobial resistant microorganisms, favor bacterial regrowth and result in changes in bacterial populations. To evaluate the feasibility of water reduction in local DWDS while ensuring water safety, water quality was measured over 2 months in two different networks, each of them harboring sub-areas with normal and reduced chlorine. Water quality remained good in chlorine reduced samples, with limited development of total flora and absence of coliforms. Furthermore, 16S rRNA amplicon-based metagenomics was used to investigate the diversity and the composition of microbial communities in the sub-networks. Taxonomic classification of sequence reads showed a reduced bacterial diversity in sampling points with higher chlorine residuals. Chlorine disinfection created more homogeneous bacterial population, dominated by Pseudomonas, a genus that contains some major opportunistic pathogens such as P. aeruginosa. In the absence of chlorine, a larger and unknown biodiversity was unveiled, also highlighted by a decreased rate of taxonomic classification to the genus and species level. Overall, this experiment in a functional DWDS will facilitate the move toward potable water delivery systems without residual disinfectants and will improve water taste for consumers.

Time series study of weather, water quality, and acute gastroenteritis at Water Safety Plan implementation sites in France and Spain.

Water Safety Plans (WSPs), recommended by the World Health Organization since 2004, can help drinking water suppliers to proactively identify potential risks and implement preventive barriers that improve safety. Few studies have investigated long-term impacts of WSPs, such as changes in drinking water quality or public health; however, some evidence from high-income countries associates WSP implementation with a reduction in diarrheal disease. To validate the previously observed linkages between WSPs and health outcomes, this time series study examined site-specific relationships between water-related exposures and acute gastroenteritis rates at three locations in France and Spain, including the role of WSP status. Relationships between control or exposure variables and health outcomes were tested using Poisson regression within generalized additive models. Controls included suspected temporal trends in disease reporting. Exposures included temperature, precipitation, raw water quality, and finished water quality (e.g., turbidity, free chlorine). In France, daily acute gastroenteritis cases were tracked using prescription reimbursements; Spanish data aggregated monthly acute gastroenteritis hospital visits. The models identified several significant relationships between indicators of exposure and acute gastroenteritis. Lag times of 6-9 days (including transit time) were most relevant for hydrological indicators (related to precipitation, runoff, and flow) at the two French sites, indicative of viral pathogens. Flush events (defined as surface runoff after a two-week antecedent dry period) linked to nonpoint source pollution were associated with a 10% increase in acute gastroenteritis rates at one location supplied by surface water. Acute gastroenteritis rates were positively associated with elevated turbidity average or maximum values in finished water at locations supplied by both surface and groundwater, by about 4% per 1-NTU increase in the two-week moving average of daily maxima or about 10% per 0.1 NTU increase in the prior month's average value. In some cases, risk appeared to be mitigated by WSP-related treatment interventions. Our results suggest drinking water exposure is associated with some potentially preventable gastrointestinal illness risk in high-income regions.

Water quality, compliance, and health outcomes among utilities implementing Water Safety Plans in France and Spain.

Water Safety Plans (WSPs), recommended by the World Health Organization since 2004, seek to proactively identify potential risks to drinking water supplies and implement preventive barriers that improve safety. To evaluate the outcomes of WSP application in large drinking water systems in France and Spain, we undertook analysis of water quality and compliance indicators between 2003 and 2015, in conjunction with an observational retrospective cohort study of acute gastroenteritis incidence, before and after WSPs were implemented at five locations. Measured water quality indicators included bacteria (E. coli, fecal streptococci, total coliform, heterotrophic plate count), disinfectants (residual free and total chlorine), disinfection by-products (trihalomethanes, bromate), aluminum, pH, turbidity, and total organic carbon, comprising about 240K manual samples and 1.2M automated sensor readings. We used multiple, Poisson, or Tobit regression models to evaluate water quality before and after the WSP intervention. The compliance assessment analyzed exceedances of regulated, recommended, or operational water quality thresholds using chi-squared or Fisher's exact tests. Poisson regression was used to examine acute gastroenteritis incidence rates in WSP-affected drinking water service areas relative to a comparison area. Implementation of a WSP generally resulted in unchanged or improved water quality, while compliance improved at most locations. Evidence for reduced acute gastroenteritis incidence following WSP implementation was found at only one of the three locations examined. Outcomes of WSPs should be expected to vary across large water utilities in developed nations, as the intervention itself is adapted to the needs of each location. The approach may translate to diverse water quality, compliance, and health outcomes.

Drinking water, diet, indoor air: Comparison of the contribution to environmental micropollutants exposure.

This study collated 254,441 analytical results from drinking water quality monitoring in order to compare levels of exposure of the French adult population from drinking water with that from total diet for 37 pesticides, 11 mineral elements, 11 polycyclic aromatic hydrocarbons (PAH), 6 non dioxin-like polychlorobiphenyls (NDL PCB), 5 ether polybromodiphenyl ethers (BDE), 2 perfluorinated compounds. It also compares levels of exposure from drinking water with that from inhalation of indoor air for 9 volatile organic compounds (VOC) and 3 phthalates. The vast majority of the water analysis results showed values below the limits of quantification and this comparison was primarily made on the basis of a highly pessimistic scenario consisting in considering the data below the limits of quantification as being equal to the limits of quantification. With this conservative scenario, it can be seen that tap water makes a minor but potentially non-negligible contribution for a few micropollutants, by comparison with diet and air. It also shows that exposure through drinking water remains below the toxicity reference values for these substances. Apart from a few extreme values reflecting exceptional local situations, the concentrations measured for the minority of positive samples (below the 95th percentile value) suggest a very low risk for human health. Lower limits of quantification would however be of use in better estimating the safety margin with regard to the toxicity reference values, in particular for BDE, PAH and NDL PCB.

Consensus report: E. coli O104:H4 (HUSEC041) and the potential threat to European water supplies.

Among the 3rd Seminar for PhD students working on Water and Health which was held in Cannes on 27­29 June 2011, experts from a number of universities and research institutes took the opportunity to discuss the emergence of Escherichia coli O104:H4 in Europe. Especially, possible threats for European water suppliers were considered. The consensus is summarized in this report. The main conclusion was that E. coli O104:H4 would not pose a substantial risk to well managed water supplies, especially where regular monitoring of indicator E. coli is negative. However, this may not apply for small and very small water systems which are quite common in Europe. New strategies like the Water Safety Plan approach are needed to protect also small scale drinking water systems and private wells in Europe. Water used in the processing of foods likely to be eaten raw, especially sprouts, should be of drinking water quality.

Biodiversity of amoebae and amoeba-associated bacteria in water treatment plants.

In this study, we enlarged our previous investigation focusing on the biodiversity of chlamydiae and amoebae in a drinking water treatment plant, by the inclusion of two additional plants and by searching also for the presence of legionellae and mycobacteria. Autochthonous amoebae were recovered onto non-nutritive agar, identified by 18S rRNA gene sequencing, and screened for the presence of bacterial endosymbionts. Bacteria were also searched for by Acanthamoeba co-culture. From a total of 125 samples, we recovered 38 amoebae, among which six harboured endosymbionts (three chlamydiae and three legionellae). In addition, we recovered by amoebal co-culture 11 chlamydiae, 36 legionellae (no L. pneumophila), and 24 mycobacteria (all rapid-growers). Two plants presented a similar percentage of samples positive for chlamydiae (11%), mycobacteria (20%) and amoebae (27%), whereas in the third plant the number of recovered bacteria was almost twice higher. Each plant exhibited a relatively high specific microbiota. Amoebae were mainly represented by various Naegleria species, Acanthamoeba species and Hartmannella vermiformis. Parachlamydiaceae were the most abundant chlamydiae (8 strains in total), and in this study we recovered a new genus-level strain, along with new chlamydiae previously reported. Similarly, about 66% of the recovered legionellae and 47% of the isolated mycobacteria could represent new species. Our work highlighted a high species diversity among legionellae and mycobacteria, dominated by putative new species, and it confirmed the presence of chlamydiae in these artificial water systems.

Free-living amoebae: Biological by-passes in water treatment.

Free-living amoebae constitute reservoirs for many bacteria including not only well-known pathogens but also emerging pathogens responsible for respiratory diseases, and contribute to the protection, survival and dissemination of these bacteria in water systems, despite the application of disinfection or thermal treatments. In this article we review the available information on the presence of free-living amoebae and amoebae-resisting bacteria in drinking water systems, on the factors that contribute to their presence in the water and/or the biofilms, on the possible control measures and their effectiveness, and we identify some gaps in current knowledge needing further research.

Novel Chlamydiales strains isolated from a water treatment plant.

Chlamydiae are obligate intracellular bacteria infecting free-living amoebae, vertebrates and some invertebrates. Novel members are regularly discovered, and there is accumulating evidence supporting a very important diversity of chlamydiae in the environment. In this study, we investigated the presence of chlamydiae in a drinking water treatment plant. Samples were used to inoculate Acanthamoeba monolayers (Acanthamoeba co-culture), and to recover autochthonous amoebae onto non-nutritive agar. Chlamydiae were searched for by a pan-chlamydia 16S rRNA gene PCR from both Acanthamoeba co-cultures and autochthonous amoebae, and phylotypes determined by 16S rRNA gene sequencing. Autochthonous amoebae also were identified by 18S rRNA gene amplification and sequencing. From a total of 79 samples, we recovered eight chlamydial strains by Acanthamoeba co-culture, but only one of 28 amoebae harboured a chlamydia. Sequencing results and phylogenetic analysis showed our strains belonging to four distinct chlamydial lineages. Four strains, including the strain recovered within its natural host, belonged to the Parachlamydiaceae; two closely related strains belonged to the Criblamydiaceae; two distinct strains clustered with Rhabdochlamydia spp.; one strain clustered only with uncultured environmental clones. Our results confirmed the usefulness of amoeba co-culture to recover novel chlamydial strains from complex samples and demonstrated the huge diversity of chlamydiae in the environment, by identifying several new species including one representing the first strain of a new family.

Biodiversity of amoebae and amoebae-resisting bacteria in a drinking water treatment plant.

The complex ecology of free-living amoebae (FLA) and their role in spreading pathogenic microorganisms through water systems have recently raised considerable interest. In this study, we investigated the presence of FLA and amoebae-resisting bacteria (ARB) at various stages of a drinking water plant fed with river water. We isolated various amoebal species from the river and from several points within the plant, mostly at early steps of water treatment. Echinamoeba- and Hartmannella-related amoebae were mainly recovered in the drinking water plant whereas Acanthamoeba- and Naegleria-related amoebae were recovered from the river water and the sand filtration units. Some FLA isolates were recovered immediately after the ozonation step, thus suggesting resistance of these microorganisms to this disinfection procedure. A bacterial isolate related to Mycobacterium mucogenicum was recovered from an Echinamoeba-related amoeba isolated from ozone-treated water. Various other ARB were recovered using co-culture with axenic Acanthamoeba castellanii, including mycobacteria, legionella, Chlamydia-like organisms and various proteobacteria. Noteworthy, a new Parachlamydia acanthamoebae strain was recovered from river water and from granular activated carbon (GAC) biofilm. As amoebae mainly multiply in sand and GAC filters, optimization of filter backwash procedures probably offers a possibility to better control these protists and the risk associated with their intracellular hosts.