Highlights
Presence of SARS-CoV-2 in feces and wastewater constitutes a paradigm shift in surveillance.
WBE has potential as a surveillance and predictive tool during the COVID-19 pandemic.
Validated protocols for concentration and quantification of SARS-CoV-2 and other markers are needed.
The presence of SARS-CoV-2 in wastewater pose the question of whether this new pandemic virus could be released into watercourses and potentially continue to finally reach coastal waters. In this study, we employed two bivalve molluscan species from the genus
Ruditapes
as sentinel organisms to investigate the presence of SARS-CoV-2 signals in the marine coastal environment. Estuarine sediments from the natural clam banks were also analyzed. Viral RNA was detected by RT-qPCR, targeting IP4, E and N1 genomic regions. Positive samples were also subjected to a PMAxx-triton viability RT-qPCR assay in order to discriminate between intact and altered capsids, obtaining indirect information about the viability of the virus. SARS-CoV-2 RNA traces were detected in 9/12 clam samples by RT-qPCR, from which 4 were positive for two different target regions. Viral quantification ranged from
Since its first identification in the United Kingdom in late 2020, the highly
transmissible B.1.1.7 variant of SARS-CoV-2 has become dominant in several countries
raising great concern. We developed a duplex real-time RT-qPCR assay to detect,
discriminate, and quantitate SARS-CoV-2 variants containing one of its mutation
signatures, the ΔHV69/70 deletion, and used it to trace the community circulation
of the B.1.1.7 variant in Spain through the Spanish National SARS-CoV-2 Wastewater
Surveillance System (VATar COVID-19). The B.1.1.7 variant was detected earlier than
clinical epidemiological reporting by the local authorities, first in the southern city
of Málaga (Andalucía) in week 20_52 (year_week), and multiple
introductions during Christmas holidays were inferred in different parts of the country.
Wastewater-based B.1.1.7 tracking showed a good correlation with clinical data and
provided information at the local level. Data from wastewater treatment plants, which
reached B.1.1.7 prevalences higher than 90% for ≥2 consecutive weeks showed that
8.1 ± 2.0 weeks were required for B.1.1.7 to become dominant. The study highlights
the applicability of RT-qPCR-based strategies to track specific mutations of variants of
concern as soon as they are identified by clinical sequencing and their integration into
existing wastewater surveillance programs, as a cost-effective approach to complement
clinical testing during the COVID-19 pandemic.
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