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

Abstract

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 faeces 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.

SARS-CoV-2 and other pathogens in municipal wastewater, landfill leachate, and solid waste: A review about virus surveillance, infectivity, and inactivation

“… Sample locations Sample types Sample storage/treatment before analysis Target genes analyzed Reference France [Paris] Wastewater samples from WWTPs. 4 °C; <24 h RdRp Wurtzer et al (2021s) India [Chennai] Samples of hospital wastewater (HWW) Ice cooled N1, N2 Chakraborty et al, 2021 India [Ahmedabad] Treated wastewater samples for receiving water bodies 4 °C; ORF1ab, N and S Kumar et al (2021) Israel (different Cites] Wastewater in its raw state from various WWTPs −80 °C or − 20 °C E Bar-Or et al, 2020 Italy [Milan and Monza] Effluent from three WWTPs and receiving rivers, both raw and treated No information ORF1ab, N, E Rimoldi et al, 2020 Italy [Milan and Rome] Raw wastewater from three WWTPs −20 °C ORF1ab, S La Rosa et al, 2020a Japan [Yamanashi] Wastewater from a single WWTP that was treated ice; <6 h N1, N2 Haramoto et al (2020) Netherlands [ Different Cities] Raw samples from WWTPs in five cities and one airport Melting ice; <24 h N1, N2, N3, E Medema et al (2020a) , b Southeastern Queensland (Australia) Raw wastewater from a pumping station and two WWTPs −80 °C; <24 h N …”

“… Environmental samples Methods Primers or probes [providers]. References Wastewater RT-qPCR N_Sarbeco [Tib-Molbiol (Berlin, Germany)] NIID_2019-nCOV_N [TOYOBO, Japan] Ahmed et al, 2000 Wastewater Sewage sludge RT-qPCR RdRp; N-gene; E-gene [Bio-Rad, Hercules, CA] Balboa et al (2020) Wastewater RT-PCR 2019-nCoV_N1–F • 2019-nCoV_N1-R • 2019-nCoV_N1–P [CDC 2019-nCoV Real-Time RT-PCR Diagnostic Panel] Medema et al (2020a) , b Wastewater RT-PCR Fast Start Universal Probe Master, forward and reverse primers including, TaqMan probe [Sigma-Aldrich] Or et al, 2020 Raw wastewater RT-qPCR nucleocapsid N gene [CDC 2019-nCoV Real-Time RT-PCR Diagnostic Panel] Wu et al (2020) Raw wastewater RT-qPCR E_Sarbeco_F • E_Sarbeco_P1 • E_Sarbeco_R [Tib-Molbiol (Berlin Germany)] Wurtzer et al (2021) …”

Wastewater monitoring as a supplementary surveillance tool for capturing SARS-COV-2 community spread. A case study in two Greek municipalities

“…Our analysis was focused on the relationship between SARS-CoV-2 RNA concentrations and 7-day cumulative cases, although a larger time window would also reflect a plausible scenario. SARS-CoV-2 RNA can enter the sewage system through wastewater discharged from households and other establishments frequently inhabited by active carriers, but also from hospitals and isolation facilities ( Giacobbo et al, 2021 ) and may occur in several forms in wastewater which include at least i) infectious protected, ii) non-infectious protected and iii) non-protected forms ( Wurtzer et al, 2021 ). It has been shown that infected persons have been found to excrete the virus for prolonged periods ( Cevik et al, 2021 , Zhang et al, 2020 ).…”

Monitoring the presence and persistence of SARS-CoV-2 in water-food-environmental compartments: State of the knowledge and research needs

“… The infectivity of both viruses was preserved up to 24 h in wastewater samples. Wurtzer et al (2021) River water and wastewater Minas Gerais State, Brazil SARS-CoV-2 SARS-CoV-2 persisted for 7.7 and 5.5 days in rain water and wastewater, respectively at 4 °C. However, the viable virus persisted more (4–4.5 times) at 24 °C in both samples.…”