Discrimination and surveillance of infectious severe acute respiratory syndrome Coronavirus 2 in wastewater using cell culture and RT-qPCR
|Elsevier - Science of the total environment (January 2022)

Link: Discrimination and surveillance of infectious severe acute respiratory syndrome Coronavirus 2 in wastewater using cell culture and RT-qPCR

# One of the a few studies that has investigated the presence of SARS-CoV-2 in treated wastewaters to determine the potential health risks across the water cycle.

# Secondary-treated wastewater (n = 89) samples were collected over a
32-week period, between April 27thand December 2nd 2020, from five
wastewater treatment plants (WWTP) located in the North of Portugal
(Vila Nova Gaia (GA) and Serzedelo (SE)) and in Lisboa e Vale do Tejo
(LVT; Alcântara (AL), Beirolas (BE), and Guia (GU)) region

# Five litre of secondary-treated wastewater were concentrated using hollow-fiber filters inuvai R180, with a molecular weight cut-off of 18.8 kDa. Samples were eluted in 300 ml buffer.

Concentration of SARS-CoV-2 from large volumes of raw wastewater is enhanced with the inuvai R180 system
|sciencedirect.com (December 2021)

The inuvai R180 Recovery Kit was validated by an independent, external Laboratory - the Water Laboratory at Instituto Superior Técnico, Lisbon, Portugal.

Link: Concentration of SARS-CoV-2 with the inuvai R180


#The inuvai R180 system had the best performance, with detection of spiked control across all samples, with average recovery percentages of 68% for porcine epidemic diarrhea virus (PEDV), with low variability

#The inuvai R180 enables the scalability of volumes without negative impact on the costs, time for analysis, and recovery/inhibition

# Hollow fiber ultrafilters favor the concentration of different microbial taxonomic groups. Such combined features make this technology attractive for usage in environmental waters monitoring.

inuvai participated with an e-poster presented by Rui Lucena at the 6th Town Hall meeting of the EU Sewage Sentinel System for SARS-CoV2 #EU4S (23-24 November 2021)

A Wastewater-based Epidemiology tool for COVID-19 Surveillance in Portugal |sciencedirect.com (October 2021)

Link: A Wastewater-based Epidemiology tool for COVID-19 Surveillance in Portugal


#SARS-CoV-2 RNA was monitored over 32-weeks in five wastewater treatment plants in Portugal, representing over two million people

# The trends of SARS-CoV-2 RNA load in raw wastewater followed the trends of daily new cases in the community, showing the usefulness of wastewater-based epidemiology as a complementary tool to clinical surveillance

Covid-19: Sewage analysis allowed to identify Covid-19 mutations in Portugal weeks before the first clinical diagnosis (May 2021)

here a summary in English of an article published by "observador" in Portuguese:

The Covidetect project revealed its first outcomes last May, 26th: it was possible to detect coronavirus mutations days or weeks in advance. It will now be used as an alert tool.

The analysis of wastewater allowed the Portuguese authorities to detect the presence of several coronavirus mutations in the Portuguese territory days or weeks before being clinically diagnosed in infected people, according to preliminary results of the Covidetect project.

The Covidetect project was launched in March 2020, the month in which the Covid-19 pandemic broke out in Portugal. A consortium led by Águas de Portugal, which includes the Faculty of Sciences of the University of Lisbon, the Instituto Superior Técnico, the General-Directorate of Health (DGS) and the Portuguese Environment Agency, with a well-defined objective: to evaluate the possibility of foreseeing the diffusion of the coronavirus in Portugal, via the analysis of sewage water.

At the genesis of the project, launched in the early days of the pandemic, is a fundamental scientific fact: an infected person (with the coronavirus) will excrete virus genetic material through feces even before having symptoms (or even not having them). That is, by analyzing the virus concentration in the various Portuguese wastewater treatment plants, it is possible to have a picture of the presence of the pandemic in a certain region of the country even before it has been triggered by the rise of number of cases.


Portugal will look for Covid-19 and new variants in sewers across the country (May 2021)

here the summary in English of an article published by "Sapo" in Portuguese:

The detection of the SARS-CoV-2 virus in wastewater allows the early identification of new outbreaks and will be extended to the whole country, with a pilot project having detected two variant mutations before the health authorities were aware of them.

The Covidetect pilot project has been running since last year and preliminary results were presented on the 26th of May in Lisbon, at the Wastewater Treatment Plant (WWTP) in Alcântara, Lisbon, with the authors explaining that the data collected in the wastewaters followed the daily evolution of the covid-19 pandemic in recent months. With the project, it was possible to detect, in Gaia, mutations of the California and Nigeria variants of the virus.

The pilot project will end in August, but the minister of Environment and Climate Change, João Pedro Matos Fernandes, who attended the ceremony, assured that the analysis of water from the WWTP will continue and expand. João Pedro Matos Fernandes stressed that not all WWTPs are prepared to carry out this detection yet, but that it is "absolutely essential" to extend the pilot project, so that water analysis becomes "a common rule for WWTPs across the country ". The WWTP that will be built or refurbished will have to be prepared for this type of analysis.

The pilot project was applied to five WWTPs in Lisbon, Cascais, Gaia and Guimarães, and the circulation of the virus in the effluent drainage networks of three reference hospitals was also monitored. Health Minister Marta Temido, who also participated in the presentation of the preliminary results of Covidetect, considered it a “powerful instrument” to fight the current pandemic and future pandemics, stressing the importance of acting as early as possible.

The next phase of the project involves disseminating the results and creating a real-time warning system to notify health and environmental authorities of the re-emergence of the virus. 760 samples of wastewater were analyzed between April 27 and December 2, 2020, "confirming that the data obtained for SARS-CoV-2 from untreated wastewater followed, in a very adjusted way, the new daily cases reported for the regions where the tested WWTP in this study are located”, said the authors.

Those responsible for the project have been collaborating with the European Commission in the EU Sewage Sentinel System for SARS-CoV-2. The Commission has already made a recommendation for a common approach to the surveillance of SARS-CoV-2 and its variants in EU wastewater. The recommendation indicates that WWTP with dimensions above 150 thousand inhabitants should be monitored.

Countries such as Australia or New Zealand are already using WWTPs as a “sentinel”, creating an 'online' system in which WWTPs appear with a green color, which turns red when genetic material from the virus is detected.


EU Recommendation COVID19 monitoring in wastewater (April 2021)

Levels of SARS-CoV-2 RNA in wastewater can be used to properly monitor COVID-19 in a community.
The European Commission recently released a recommendation on a common approach to establish a systematic surveillance
of SARS-CoV-2 and its variants in wastewaters in the EU.
The monitoring system should include at least wastewaters from large cities with over 150000 inhabitants, preferably with a minimum sampling frequency of two samples per week.
Effective concentration method is needed for recovery of SARS-CoV-2 from raw wastewater.
The inuvai R180 Recovery Kit offers a technically simple and fast method, with high recovery of target microorganisms, in a reduced final volume concentrate from a large volume sample.

link : EU recommendation covid19 monitoring wastewater

Researchers look for traces of coronavirus in wastewater

The video from the Portuguese news channel SIC was filmed in the Water Laboratory of the Instituto Superior Técnico, showing the inuvai R180 Recovery Kit in use. The R180 concentration method goes beyond SARS CoV-2 and can be applied to the recovery of other microorganisms making it a useful tool for a multi-pathogen approach, from both wastewater-based epidemiology and water quality monitoring perspectives.

The video from the Portuguese News Channel SIC (audio in Portuguese language):