A Flexible Smoother Adapted to Censored Data With Outliers and Its Application to SARS-CoV-2 Monitoring in Wastewater

A sentinel network, Obépine, has been designed to monitor SARS-CoV-2 viral load in wastewaters arriving at wastewater treatment plants (WWTPs) in France as an indirect macro-epidemiological parameter. The sources of uncertainty in such a monitoring system are numerous, and the concentration measurements it provides are left-censored and contain outliers, which biases the results of usual smoothing methods. Hence, the need for an adapted pre-processing in order to evaluate the real daily amount of viruses arriving at each WWTP. We propose a method based on an auto-regressive model adapted to censored data with outliers. Inference and prediction are produced via a discretized smoother which makes it a very flexible tool. This method is both validated on simulations and real data from Obépine. The resulting smoothed signal shows a good correlation with other epidemiological indicators and is currently used by Obépine to provide an estimate of virus circulation over the watersheds corresponding to about 200 WWTPs. Copyright © 2022 Courbariaux, Cluzel, Wang, Maréchal, Moulin, Wurtzer, Obépine Consortium, Mouchel, Maday and Nuel.

OBEPINE: A FRENCH EXPERIMENT TO MONITOR THE COVID-19 EPIDEMIC THROUGH WASTEWATER

Since March 2020, the Obépine (Observatoire Epidémiologique dans les Eaux Usées) research consortium has been working on exploiting the presence of SARS-CoV-2 in wastewater to propose epidemiological monitoring of COVID-19 in France. Although wastewater epidemiology is a long-standing discipline, it has never been so widely deployed on a national and international scale. Bringing together research teams with varied skills, Obépine has established the first protocols for quantifying the viral genome in samples from wastewater treatment plants (WWTPs). The sentinel network, which was built with the support of the French Ministry of Higher Education, Research and Innovation, currently includes 200 WWTPs in mainland France and overseas. It allows the bi-weekly assessment of the dynamics of the epidemic in more than 33% of the French population. This paper presents the genesis of a surveillance project and the lines of research of a network that wishes to work - in the near future - to complete the existing early warning and monitoring systems for endemic or emerging infections. © 2021 Academie Veterinaire de France. All rights reserved.

SARS-CoV-2 genome quantification in wastewaters at regional and city scale allows precise monitoring of the whole outbreaks dynamics and variants spreading in the population.

SARS-CoV-2 is a coronavirus causing a globalized outbreak called COVID-19. SARS-CoV-2 transmission is associated with inhalation of contaminated respiratory droplets and could causes severe complications. Until today several "waves" of infections have been observed despite implementation of strict health policies. Decisions for such sanitary measures are based on population health monitoring. Unfortunately, for COVID-19, a significant proportion of individuals are asymptomatic but play a role in the virus transmission. To overcome these limitations, several strategies were developed including genome quantification in wastewater that could allow monitoring of the health status of population, since shedding of SARS-CoV-2 in patient stool is frequent. Wastewater-based epidemiology (WBE) was established and several countries implemented this approach to allow COVID-19 outbreak monitoring. In France, the OBEPINE project performed a quantitative analysis of SARS-CoV-2 in raw wastewater samples collected from major wastewater treatment plants (WWTP) since March 2020. In the greater Paris area 1101 samples (507 for five WWTP and 594 for sewer) were collected. This 16 months monitoring allows us to observe the outbreak dynamics. Comparison of WBE indicators with health data lead to several important observation; the good level of correlation with incidence rates, the average 3 days lead time, and the sensitivity (WBE change when incidence is > to 7/100000 inhabitants). We also compared the local monitoring (city level) with the regional monitoring, to help cluster identification. Moreover, variants of concern (VOC) emerged due to the selection pressure. We developed a specific RT-qPCR method targeting the deletion H69-V70 in the spike protein, using this deletion as a proxy of the B.1.1.7 presence in the wastewater. With this data we demonstrate the predominant role played by this strain in the third wave. All these results allow a better description and understanding of the pandemic and highlight the role of such WBE indicators.

Several forms of SARS-CoV-2 RNA can be detected in wastewaters: Implication for wastewater-based epidemiology and risk assessment.

The ongoing global pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a public health emergency of international concern. Although SARS-CoV-2 is considered to be mainly transmitted by inhalation of contaminated droplets and aerosols, SARS-CoV-2 is also detected in human feces and to a less extent in urine, and in raw wastewaters (to date viral RNA only) suggesting that other routes of infection may exist. Monitoring SARS-CoV-2 genomes in wastewaters has been proposed as a complementary approach for tracing the dynamics of virus transmission within human population connected to wastewater network. The understanding on SARS-CoV-2 transmission through wastewater surveillance, the development of epidemic modeling and the evaluation of SARS-CoV-2 transmission from contaminated wastewater are largely limited by our knowledge on viral RNA genome persistence and virus infectivity preservation in such an environment. Using an integrity based RT-qPCR assay this study led to the discovery that SARS-CoV-2 RNA can persist under several forms in wastewaters, which provides important information on the presence of SARS-CoV-2 in raw wastewaters and associated risk assessment.

Evaluation of lockdown effect on SARS-CoV-2 dynamics through viral genome quantification in waste water, Greater Paris, France, 5 March to 23 April 2020.

IntroductionSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of coronavirus disease (COVID-19). People infected with SARS-CoV-2 may exhibit no or mild non-specific symptoms; thus, they may contribute to silent circulation of the virus among humans. Since SARS-CoV-2 RNA can be detected in stool samples, monitoring SARS-CoV-2 RNA in waste water (WW) has been proposed as a complementary tool to investigate virus circulation in human populations.AimTo test if the quantification of SARS-CoV-2 genomes in WW correlates with the number of symptomatic or non-symptomatic carriers.MethodWe performed a time-course quantitative analysis of SARS-CoV-2 by RT-qPCR in raw WW samples collected from several major WW treatment plants in Greater Paris. The study period was 5 March to 23 April 2020, including the lockdown period in France (from 17 March).ResultsWe showed that the increase of genome units in raw WW accurately followed the increase of human COVID-19 cases observed at the regional level. Of note, the viral genome could be detected before the epidemic grew massively (around 8 March). Equally importantly, a marked decrease in the quantities of genome units was observed concomitantly with the reduction in the number of new COVID-19 cases, 29 days following the lockdown.ConclusionThis work suggests that a quantitative monitoring of SARS-CoV-2 genomes in WW could generate important additional information for improved monitoring of SARS-CoV-2 circulation at local or regional levels and emphasises the role of WW-based epidemiology.