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 (R) 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 (R) 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.

A Methodological Approach to Water Concentration to Investigate the Presence of SARS-CoV-2 RNA in Surface Freshwaters / M. Tesauro, M. Terraneo, M. Consonni, C. Fappani, D. Colzani, C. Stevanin, A. Amendola, D. Masseroni, E. Tanzi. - In: PATHOGENS. - ISSN 2076-0817. - 11:8(2022), pp. 845.1-845.15. [10.3390/pathogens11080845]

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

M. Tesauro
Primo
;
M. Terraneo
Secondo
;
M. Consonni;C. Fappani;D. Colzani;A. Amendola;D. Masseroni
Penultimo
;
E. Tanzi
Ultimo
2022

Abstract

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 (R) 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 (R) 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.
SARS-CoV-2; flood spillways; surface freshwater
Settore MED/42 - Igiene Generale e Applicata
Settore AGR/08 - Idraulica Agraria e Sistemazioni Idraulico-Forestali
CAR_EXT20ETANZ_01 - Studio sulla presenza di materiale genetico di SARS-CoV-2 nelle acque superficiali dei corpi idrici Ricettori che ricevono le acque di Sfioro provenienti dalle reti di drenaggio urbano (QuoViRuS) - MASSERONI, DANIELE - CAR_EXT - CARIPLO - EXTRABANDO E PROGETTI TERRITORIALI - 2020
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/947056
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