BA.2.86 variant emergence and spread dynamics through wastewater monitoring in Paris, France.

Numerous SARS-CoV-2 variants are emerging as the epidemic continues, inducing new waves of contamination. In July 2023, a new variant named BA.2.86 was identified, raising concerns about its potential for viral escape, even in an immune population. The reduction in patient-centered testing and the identification of variants by sequencing means that we are now blind to the spread of this new variant. The aim of this study was to track the signature of this variant in wastewater in Paris, France. This variant showed a very rapid spread, highly correlated with national flash studies involving sequencing of clinical samples, but with a moderate impact on virus circulation. This easy-to-implement approach enabled us to monitor the emergence and spread of this new variant in real time at low cost.

From Alpha to Omicron BA.2: New digital RT-PCR approach and challenges for SARS-CoV-2 VOC monitoring and normalization of variant dynamics in wastewater.

Throughout the COVID-19 pandemic, new variants have continuously emerged and spread in populations. Among these, variants of concern (VOC) have been the main culprits of successive epidemic waves, due to their transmissibility, pathogenicity or ability to escape the immune response. Quantification of the SARS-CoV-2 genomes in raw wastewater is a reliable approach well-described and widely deployed worldwide to monitor the spread of SARS-CoV-2 in human populations connected to sewage systems. Discrimination of VOCs in wastewater is also a major issue and can be achieved by genome sequencing or by detection of specific mutations suggesting the presence of VOCs. This study aimed to date the emergence of these VOCs (from Alpha to Omicron BA.2) by monitoring wastewater from the greater Paris area, France, but also to model the propagation dynamics of these VOCs and to characterize the replacement kinetics of the prevalent populations. These dynamics were compared to various individual-centered public health data, such as regional incidence and the proportions of VOCs identified by sequencing of strains isolated from patient. The viral dynamics in wastewater highlighted the impact of the vaccination strategy on the viral circulation within human populations but also suggested its potential effect on the selection of variants most likely to be propagated in immunized populations. Normalization of concentrations to capture population movements appeared statistically more reliable using variations in local drinking water consumption rather than using PMMoV concentrations because PMMoV fecal shedding was subject to variability and was not sufficiently relevant in this study. The dynamics of viral spread was observed earlier (about 13 days on the wave related to Omicron VOC) in raw wastewater than the regional incidence alerting to a possible risk of decorrelation between incidence and actual virus circulation probably resulting from a lower severity of infection in vaccinated populations.

A nationwide indicator to smooth and normalize heterogeneous SARS-CoV-2 RNA data in wastewater.

Since many infected people experience no or few symptoms, the SARS-CoV-2 epidemic is frequently monitored through massive virus testing of the population, an approach that may be biased and may be difficult to sustain in low-income countries. Since SARS-CoV-2 RNA can be detected in stool samples, quantifying SARS-CoV-2 genome by RT-qPCR in wastewater treatment plants (WWTPs) has been carried out as a complementary tool to monitor virus circulation among human populations. However, measuring SARS-CoV-2 viral load in WWTPs can be affected by many experimental and environmental factors. To circumvent these limits, we propose here a novel indicator, the wastewater indicator (WWI), that partly reduces and corrects the noise associated with the SARS-CoV-2 genome quantification in wastewater (average noise reduction of 19%). All data processing results in an average correlation gain of 18% with the incidence rate. The WWI can take into account the censorship linked to the limit of quantification (LOQ), allows the automatic detection of outliers to be integrated into the smoothing algorithm, estimates the average measurement error committed on the samples and proposes a solution for inter-laboratory normalization in the absence of inter-laboratory assays (ILA). This method has been successfully applied in the context of Obépine, a French national network that has been quantifying SARS-CoV-2 genome in a representative sample of French WWTPs since March 5th 2020. By August 26th, 2021, 168 WWTPs were monitored in the French metropolitan and overseas territories of France. We detail the process of elaboration of this indicator, show that it is strongly correlated to the incidence rate and that the optimal time lag between these two signals is only a few days, making our indicator an efficient complement to the incidence rate. This alternative approach may be especially important to evaluate SARS-CoV-2 dynamics in human populations when the testing rate is low.

First evidence of SARS-CoV-2 genome detection in zebra mussel (Dreissena polymorpha).

The uses of bivalve molluscs in environmental biomonitoring have recently gained momentum due to their ability to indicate and concentrate human pathogenic microorganisms. In the context of the health crisis caused by the COVID-19 epidemic, the objective of this study was to determine if the SARS-CoV-2 ribonucleic acid genome can be detected in zebra mussels (Dreissena polymorpha) exposed to raw and treated urban wastewaters from two separate plants to support its interest as bioindicator of the SARS-CoV-2 genome contamination in water. The zebra mussels were exposed to treated wastewater through caging at the outlet of two plants located in France, as well as to raw wastewater in controlled conditions. Within their digestive tissues, our results showed that SARS-CoV-2 genome was detected in zebra mussels, whether in raw and treated wastewaters. Moreover, the detection of the SARS-CoV-2 genome in such bivalve molluscans appeared even with low concentrations in raw wastewaters. This is the first detection of the SARS-CoV-2 genome in the tissues of a sentinel species exposed to raw and treated urban wastewaters. Despite the need for development for quantitative approaches, these results support the importance of such invertebrate organisms, especially zebra mussel, for the active surveillance of pathogenic microorganisms and their indicators in environmental waters.

Multiresidue Methods for the Determination of Organic Micropollutants and Their Metabolites in Fish Matrices.

Two analytical methods were developed for the determination of 48 organic compounds and 20 of their main by-products in fish matrices. The targeted compounds belong to various chemical classes of metabolizable (phthalates, polycyclic aromatic hydrocarbons, insecticides [pyrethroids and N,N-diethyl-meta-toluamide]) and legacy (organochlorine pesticides, polychlorinated biphenyls, polybrominated diphenyl ethers) pollutants. Analyses were performed by gas and liquid chromatography-tandem mass spectrometry in multiple reaction monitoring (MRM) and dynamic MRM, respectively. Method performances were satisfactory, with results meeting the validation criteria because they achieved good linearity responses, recovery, precision, and accuracy for most of the 68 investigated compounds. The methods were then applied on 3 feral chub (Squalius cephalus) collected from the Marne hydrographic network (France). Twenty-six parent compounds and 5 metabolites were systematically detected in fish matrices, with substantial concentration variability within and among individuals. Phthalates and pyrethroids accounted for most of the pollutant load. Metabolite concentrations in liver samples exceeded those of parent molecules in fish muscle. The present study presents 2 reliable methods for the determination of a wide range of contaminants and underlines the importance of metabolite analysis for a more comprehensive understanding of pollutant bioaccumulation and fate in aquatic organisms. Environ Toxicol Chem 2019;38:1866-1878. © 2019 SETAC.

Investigating the metal contamination of sediment transported by the 2016 Seine River flood (Paris, France).

Fine sediment transport in rivers is exacerbated during flood events. These particles may convey various contaminants (i.e. metals, pathogens, industrial chemicals, etc.), and significantly impact water quality. The exceptional June 2016 flood of the Seine River (catchment area: 65 000 km2, France), potentially mobilized and deposited contaminated materials throughout the Paris region. Flood sediment deposits (n = 29) were collected along the Seine River and its main tributaries upstream (Yonne, Loing and Marne Rivers) and downstream of Paris (Oise and Eure Rivers). Fallout radionuclides (137Cs, 7Be) were measured to characterize the sources of the material transiting the river, while trace elements (e.g. Cr, Ni, Zn, Cu, As, Cd, Sb, Pb, Tl, Ag) and stable lead isotopes (206Pb/207Pb) were analyzed to quantify the contamination of sediment transported during the flood. In upper sections of the Seine River, sediment mainly originated from the remobilization of particles with a well-balanced contribution of surface and subsurface sources. In the upstream tributaries, sediment almost exclusively originated from the remobilization of subsurface particles. In Paris and downstream of Paris, recently eroded particles and surface sources dominated, suggesting particles were mainly supplied by urban runoff and the erosion of agricultural soils. The highest metal concentrations and Enrichment Factors (EF) were found in the sediment collected in the Loing, Orge and Yvette upstream tributaries. Although these inputs were diluted in the Seine River, an increase in elemental concentrations was observed, progressing downstream through Paris. However, EFs in sediment collected along the Seine River were lower or in the same range of values sampled over the last several decades, reflecting the progressive decontamination of the urbanized Seine River basin.

Nitrite reduction by biogenic hydroxycarbonate green rusts: evidence for hydroxy-nitrite green rust formation as an intermediate reaction product.

The present study investigates for the first time the reduction of nitrite by biogenic hydroxycarbonate green rusts, bio-GR(CO3), produced from the bioreduction of ferric oxyhydroxycarbonate (Fohc), a poorly crystalline solid phase, and of lepidocrocite, a well-crystallized Fe(III)-oxyhydroxide mineral. Results show a fast Fe(II) production from Fohc, which leads to the precipitation of bio-GR(CO3) particles that were roughly 2-fold smaller (2.3 ± 0.4 µm) than those obtained from the bioreduction of lepidocrocite (5.0 ± 0.4 µm). The study reveals that both bio-GR(CO3) are capable of reducing nitrite ions into gaseous nitrogen species such as NO, N2O, or N2 without ammonium production at neutral initial pH and that nitrite reduction proceeded to a larger extent with smaller particles than with larger ones. On the basis of the identification of intermediates and end-reaction products using X-ray diffraction and X-ray absorption fine structure (XAFS) spectroscopy at the Fe K-edge, our study shows the formation of hydroxy-nitrite green rust, GR(NO2), a new type of green rust 1, and suggests that the reduction of nitrite by biogenic GR(CO3) involves both external and internal reaction sites and that such a mechanism could explain the higher reactivity of green rust with respect to nitrite, compared to other mineral substrates possessing only external reactive sites.

Combining measurements and modelling to quantify the contribution of atmospheric fallout, local industry and road traffic to PAH stocks in contrasting catchments.

Various sources supply PAHs that accumulate in soils. The methodology we developed provided an evaluation of the contribution of local sources (road traffic, local industries) versus remote sources (long range atmospheric transport, fallout and gaseous exchanges) to PAH stocks in two contrasting subcatchments (46-614 km²) of the Seine River basin (France). Soil samples (n = 336) were analysed to investigate the spatial pattern of soil contamination across the catchments and an original combination with radionuclide measurements provided new insights into the evolution of the contamination with depth. Relationships between PAH concentrations and the distance to the potential sources were modelled. Despite both subcatchments are mainly rural, roadside areas appeared to concentrate 20% of the contamination inside the catchment while a local industry was found to be responsible for up to 30% of the stocks. Those results have important implications for understanding and controlling PAH contamination in rural areas of early-industrialized regions.

Variation of raw wastewater microbiological quality in dry and wet weather conditions.

The microbiological quality of urban wastewaters presents important environmental, sanitary, and political challenges. However, the variability of untreated wastewater quality is seldom known when it comes to microbial parameters. This study aims to evaluate the variability of microbiological quality in wastewater influents from different wastewater treatment plants connected to combined and partially separate sewer networks in the Parisian area and to evaluate the impact of this variability on the treatment efficiency and on the accuracy of wastewater effluent monitoring. The densities of fecal indicator bacteria (FIB), Escherichia coli and intestinal enterococci, and their partitioning on settleable particles were analyzed at the inlet of two wastewater treatment plants during dry weather (130 composite samples and 7 days sampled every 2 hours) and storm events (39 composite samples, and 7 rain courses) from 2008 to 2012. The results showed that fecal indicator densities vary according to the network characteristics and according to the meteorological conditions. During storm events, a significant dilution of E. coli and enterococci was observed, as well as a decrease in the settleable fraction of E. coli during the maximal impact of the storm. However, storm events did not significantly impact the regular FIB monitoring. FIB removals by primary and secondary treatment were significantly correlated with FIB densities in influent wastewater; however, meteorological conditions also influenced the removal of FIB.

Mass balance and decontamination times of Polycyclic Aromatic Hydrocarbons in rural nested catchments of an early industrialized region (Seine River basin, France).

Accumulation of Polycyclic Aromatic Hydrocarbons (PAHs) in soils and their subsequent release in rivers constitute a major environmental and public health problem in industrialized countries. In the Seine River basin (France), some PAHs exceed the target concentrations, and the objectives of good chemical status required by the European Water Framework Directive might not be achieved. This investigation was conducted in an upstream subcatchment where atmospheric fallout (n=42), soil (n=33), river water (n=26) and sediment (n=101) samples were collected during one entire hydrological year. PAH concentrations in atmospheric fallout appeared to vary seasonally and to depend on the distance to urban areas. They varied between 60 ng·L(-1) (in a remote site during autumn) and 2,380 ng·L(-1) (in a built-up area during winter). PAH stocks in soils of the catchment were estimated based on land use, as mean PAH concentrations varied between 110 ng·g(-1) under woodland and 2,120 ng·g(-1) in built-up areas. They ranged from 12 to 220 kg·km(-2). PAH contamination in the aqueous phase of rivers remained homogeneous across the catchment (72 ± 38 ng·L(-1)). In contrast, contamination of suspended solid was heterogeneous depending on hydrological conditions and population density in the drainage area. Moreover, PAH concentrations appeared to be higher in sediment (230-9,210 ng·g(-1)) than in the nearby soils. Annual mass balance calculation conducted at the catchment scale showed that current PAH losses were mainly due to dissipation (biodegradation, photo-oxidation and volatilization) within the catchments (about 80%) whereas exports due to soil erosion and riverine transport appeared to be of minor importance. Based on the calculated fluxes, PAHs appeared to have long decontamination times in soils (40 to 1,850 years) thereby compromising the achievement of legislative targets. Overall, the study highlighted the major role of legacy contamination that supplied the bulk of PAHs that are still found nowadays in the environment.

Spatio-temporal variability of solid, total dissolved and labile metal: passive vs. discrete sampling evaluation in river metal monitoring.

In order to obtain representative dissolved and solid samples from the aquatic environment, a spectrum of sampling methods are available, each one with different advantages and drawbacks. This article evaluates the use of discrete sampling and time-integrated sampling in illustrating medium-term spatial and temporal variation. Discrete concentration index (CI) calculated as the ratio between dissolved and solid metal concentrations in grab samples are compared with time-integrated concentration index (CI) calculated from suspended particulate matter (SPM) collected in sediment traps and labile metals measured by the diffusive gel in thin films (DGT) method, collected once a month during one year at the Seine River, upstream and downstream of the Greater Paris Region. Discrete CI at Bougival was found to be significantly higher than at Triel for Co, Cu, Mn, Ni and Zn, while discrete metal partitioning at Marnay was found to be similar to Bougival and Triel. However, when using time-integrated CI, not only was Bougival CI significantly higher than Triel CI, CI at Marnay was also found to be significantly higher than CI at Triel which was not observed for discrete CI values. Since values are time-averaged, dramatic fluctuations were smoothed out and significant medium-term trends were enhanced. As a result, time-integrated concentration index (CI) was able to better illustrate urbanization impact between sites when compared to discrete CI. The impact of significant seasonal phenomenon such as winter flood, low flow and redox cycles was also, to a certain extent, visible in time-integrated CI values at the upstream site. The use of time-integrated concentration index may be useful for medium- to long-term metal studies in the aquatic environment.

Impact of an intense combined sewer overflow event on the microbiological water quality of the Seine River.

For a better understanding of the short and mid-term impacts of a combined sewer overflow (CSO) on the microbiological quality of the receiving river, we studied the composition of a CSO discharge and monitored during several hours the changes in the concentration of fecal indicator bacteria (FIB) in the impacted river water mass. The CSO occurred at the Clichy outfall (Paris agglomeration, France) in summer 2008 as a result of the most intense rainfall of the year. In 6h, 578, 705 m(3) of sewage and 124 t of suspended matter (SM) were discharged into the Seine River. The CSO contained 1.5 × 10(6)E. coli and 4.0 × 10(5) intestinal enterococci per 100 mL on average, and 77% of the E. coli were attached to SM. It was estimated that 89% of the CSO discharge was contributed by surface water runoff, and that resuspension of sewer sediment contributed to ∼75% of the SM, 10-70% of the E. coli and 40-80% of the intestinal enterococci. Directly downstream from the CSO outfall, FIB concentrations in the impacted water mass of the Seine River (2.9 × 10(5)E. coli and 7.6 × 10(4) intestinal enterococci per 100 mL) exceeded by two orders of magnitude the usual dry weather concentrations. After 13-14 h of transit, these concentrations had decreased by 66% for E. coli and 79% for intestinal enterococci. This decline was well accounted for by our estimations of dilution, decay resulting from mortality or loss of culturability and sedimentation of the attached fraction of FIB.

Combined monitoring of changes in delta13CH4 and archaeal community structure during mesophilic methanization of municipal solid waste.

Reconstituted municipal solid waste (MSW) with varying contents of putrescible and cellulosic waste was incubated anaerobically under mesophilic conditions. Standard physicochemical parameters were monitored, together with stable isotopic signatures of produced CH(4) and CO(2). delta(13)C values for CH(4) indicated a change of methanogenic metabolism with time. CH(4) was predominantly produced from H(2)/CO(2) at the beginning of the incubations. This period was associated with important shifts in archaeal communities monitored by automated ribosomal intergenic spacer analysis (ARISA) and FISH of oligonucleotidic probes targeting specifically 16S rRNA gene of various methanogenic groups. The onset of the active methane generation phase was characterized by an increase of CH(4)delta(13)C, indicating a progressive shift toward an aceticlastic metabolism. When the methane production levelled off, a decrease in the isotopic signature was observed toward values characteristics of hydrogenotrophic metabolism. ARISA profiles were, however, found to be stable from the beginning of the active methane generation phase until the end of the experiment. FISH observation indicated that members of the family Methanosarcinaceae were predominant in the archaeal community during this period, suggesting that these methanogens might exhibit a high metabolic versatility during methanization of waste.

Efficiency evaluation of sewage treatment plants with different technologies in Delhi (India).

Physical, chemical and microbiological efficiencies of Sewage Treatment Plants (STPs) located in Delhi's watershed in context of different treatment technologies employed in these plants have been determined. There were in all seventeen STPs treating domestic wastewater which were studied over a period of 12 months. These STPs were based on Conventional Activated sludge process (ASP), Extended aeration (Ex. Aeration), physical, chemical and biological removal treatment (BIOFORE) and oxidation pond treatment process. Results suggests that except "Mehrauli" STP which was based on Extended aeration process and "Oxidation pond", effluents from all other STPs exceeded FC standard of 10(3) MPN/100 ml for unrestricted irrigation criteria set by National river conservation directorate (NRCD). Actual integrated efficiency (IE(a)) of each STP was evaluated and compared with the standard integrated efficiency (IE(s)) based upon physical, biological and microbiological removal efficiencies depending upon influent sewage characteristics. The best results were obtained for STPs employing extended aeration, BIOFORE and oxidation pond treatment process thus can be safely used for irrigation purposes.

Dissolved organic matter from treated effluent of a major wastewater treatment plant: characterization and influence on copper toxicity.

A combination of reverse osmosis (RO) concentration and DAX-8/XAD-4 resin adsorption techniques is used to isolate the various constituents of urban dissolved organic matter (DOM) from inorganic salts. Three fractions: hydrophobic (HPO), transphilic (TPI) and hydrophilic (HPI) accounting respectively for 35%, 20% and 45% of extracted carbon, are isolated from effluents of a major French wastewater treatment plant. This atypical DOC distribution, in comparison with natural water where the HPO fraction dominates, shows the significance of HPI fraction which often gets neglected because of extraction difficulties. A number of analytical techniques (elemental, spectroscopic: UV, FTIR) allow highlighting the weak aromaticity of wastewater effluent DOM (EfOM) due to fewer degradation and condensation processes and the strong presence of proteinaceous structures indicative of intense microbial activity. Copper toxicity in the presence of DOM is estimated using an acute toxicity test on Daphnia Magna (Strauss). Results reveal the similar protective role of each EfOM fraction compared to reference Suwannee river fulvic acid despite lower EfOM aromaticity (i.e. specific UV absorbance). The environmental implications of these results are discussed with respect to the development of site-specific water quality criteria.

Dissolved and bioavailable contaminants in the Seine river basin.

Diffusive Gradient in Thin Films (DGT) and Semi-Permeable Membrane Devices (SPMDs) were deployed in the Seine river basin in order to assess labile metals and truly dissolved Polycyclic Aromatic Hydrocarbons. We show that the tools are reliable in aquatic environments to assess the speciation of dissolved contaminants and hence provide a good insight into the potential bioavailability of contaminants. The deployment of the DGT and SPMDs in contrasting environments in the Seine river basin allowed distinction to be made of availability of contaminants between headwater streams and much more impacted river reaches and an assessment of bioavailability. At the stations under urban influence, the impact of dissolved organic matter on both copper and PAHs bioavailability is less pronounced than at upstream stations, where humic substances dominate.

Modelling the impacts of Combined Sewer Overflows on the river Seine water quality.

To achieve the objectives of the European Water Framework Directive (EWFD), the Seine basin Water Authority has constructed a number of prospective scenarios forecasting the impact of planned investments in water quality. Paris and its suburbs were given special attention because of their impact on the river Seine. Paris sewer system and overflow control is of major concern in future management plans. The composition and fate of the urban effluents have been characterized through numerous in situ samplings, laboratory experiments and modelling studies. The PROSE model was especially designed to simulate the impact on the river of both permanent dry-weather effluents and of highly transient Combined Sewer Overflow (CSO). It was also used to represent the impact of Paris at large spatial and temporal scales. In addition to immediate effects on oxygen levels, heavy particulate organic matter loads that settle downstream of the outlets contribute to permanent oxygen consumption. Until the late 90s, the 50 km long reach of the Seine inside Paris was permanently affected by high oxygen consumption accounting for 112% of the flux upstream of the city. 20% of this demand resulted from CSO. However, the oxygenation of the system is strong due to high phytoplankton activity. As expected, the model results predict a reduction of both permanent dry-weather effluents and CSOs in the future that will greatly improve the oxygen levels (concentrations higher than 7.3 mgO(2) L(-1), 90% of the time instead of 4.0 mgO(2) L(-1) in the late 90s). The main conclusion is that, given the spatial and temporal extent of the impact of many CSOs, water quality models should take into account the CSOs in order to be reliable.

Distribution of 7Be, 210Pb and 137Cs in watersheds of different scales in the Seine River basin: inventories and residence times.

The activity of environmental radionuclides ((7)Be, (210)Pb and (137)Cs) was monitored in nested catchments, inside the Seine River basin. Suspended matter data was collected at 8 different watersheds, ranging from order 1 to order 7, and ranging in size over 4 orders of magnitude. Suspended matter was analyzed for (210)Pb, (137)Cs and (7)Be, and used to calculate the flux of sediments out of each watershed. Monthly atmospheric flux data of (210)Pb and (7)Be was analyzed to assess the input flux of each into the watersheds, taking into account the rainfall during sampling periods. Taking advantage of the different half-lives of (7)Be (53 days) and (210)Pb (22 years), a two-box model was built for each of the catchments following a methodology previously developed by Dominik et al. [Dominik J, Burrus D, Vernet JP. Transport of the environmental radionuclides in alpine watershed. Earth Planet Sci Letters 1987; 84: 165-180.]. The model divides the watershed into a soil box and a rapid reservoir and provides insight into the removal rate of suspended matter from the surrounding watershed. The model enables the assessment of the surface area and the residence time of slow and rapid reservoirs to describe the fate of contaminants of atmospheric origin inside the river basin. The model was improved by considering the dissolved fraction in the total flux and adding the (137)Cs inventory as an additional constraint. The effects of these changes are discussed. Residence times in the soil box, characterized by low transport velocity, range between 4800 years at Melarchez (order 1) to about 30000 years at Andresy and Poses (order 7). They remain constant in each watershed over a large range of variation of atmospheric fluxes of (7)Be and (210)Pb during the whole study, but are sensitive to SM variations. The residence time in the rapid box, which includes the surface of the river and immediate surroundings, is less than one year, while its surface area is in the range 0.6% to 2.2% of the total catchment area. They are sensitive to (7)Be atmospheric flux variations. The two-box model was used to estimate the amount of the radionuclides in each reservoir. Inventories appear to be constant from one watershed to the next. The (7)Be inventory ratio in the rapid and slow boxes expresses the rate of particle-reactive atmospheric pollutants that will be rapidly delivered to the river.