A Methodological Approach to Water Concentration to Investigate the Presence of SARS-CoV-2 RNA in Surface Freshwaters

A Methodological Approach to Water Concentration to Investigate the Presence of SARS-CoV-2 RNA in Surface Freshwaters

During the COVID-19 public health emergency, an increasing number of studies reported the occurrence of SARS-CoV-2 in wastewaters worldwide, but little is known about the presence of the virus in surface freshwaters. The aim of the current study was to develop and validate an appropriate and scalabl...

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Bibliographic Details
Published in:Pathogens (Basel) Vol. 11; no. 8; p. 845
Main Authors: Tesauro, Marina, Terraneo, Mara, Consonni, Michela, Fappani, Clara, Colzani, Daniela, Stevanin, Caterina, Amendola, Antonella, Masseroni, Daniele, Tanzi, Elisabetta
Format: Journal Article
Language:English
Published: Switzerland MDPI AG 28-07-2022
MDPI
Subjects:
Acids
Aerosols
Canals
Climatic changes
Contamination
Coronaviruses
COVID-19
Dextran
Dextrans
Disease transmission
Distilled water
Environmental aspects
Experiments
Filter paper
Filtration
flood spillways
Fresh water
Health aspects
Methods
Nucleic acids
Public health
Recovery
SARS-CoV-2
Severe acute respiratory syndrome coronavirus 2
Silica
Spillways
Stormwater management
surface freshwater
Surface water
Ultrafiltration
Viral diseases
Viruses
Water analysis
Water monitoring
Water purification
Water sampling
Italy
surface freshwater
SARS-CoV-2
flood spillways
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Summary:During the COVID-19 public health emergency, an increasing number of studies reported the occurrence of SARS-CoV-2 in wastewaters worldwide, but little is known about the presence of the virus in surface freshwaters. The aim of the current study was to develop and validate an appropriate and scalable methodological approach for the concentration and detection of SARS-CoV-2 from surface freshwater samples, collected within the Milan rural network subjected to flood spillways activity. Overall, both surface water and distilled water samples spiked with inactivated SARS-CoV-2 were used to validate the concentration method for pathogens determination. Two pre-filtration systems, filter paper and Sartolab P20 (Sartorius, Germany) and two concentration methods, two-phase (PEG-dextran method) separation and tangential flow ultrafiltration (UF), were compared. The effects of pre-filtration and concentration on viral nucleic acid recovery were assessed through real time RT-PCR targeting SARS-CoV-2 and the internal viral control PMMoV (Pepper Mild Mottle Virus). Our results showed that UF is more sensitive than the PEG-dextran method in viral acid nucleic recovery from surface water samples. Better results were obtained pre-filtering samples with Sartolab P20 and extracting the nucleic acids with undiluted silica, rather than diluted as required by the standard protocol. The proposed method will be used for the monitoring of surface waters in the Milan area.
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ISSN:2076-0817
2076-0817
DOI:10.3390/pathogens11080845