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A study on the presence of SARS-CoV-2, rotavirus A, norovirus and hepatitis E virus in Croatian environment

The SARS-CoV-2 pandemic has put enormous pressure on health services worldwide, while enteric viruses pose a continuing public health threat. The latter are represented mainly by rotavirus A (RVA), norovirus (NoV) and hepatitis E (HEV) virus strains. The research on the environmental presence of these RNA viruses is an important early indicator of possible outbreaks in the community. The aim of this study was to investigate the presence of SARS-CoV-2, RVA, NoV and HEV in the environment in Croatia. During two seasons (2018/2019, 2019/2020) we have collected 48 surface and sewage water samples originating from five locations in continental Croatia. Additionally we have sampled three species of bivalves (Mytilus galloprovincialis, Ostrea edulis, Venus verrucosa) from the Croatian side of the Adriatic coast, that yielded 198 hepatopankreas samples. The virus concentration procedure was performed according to the standard ISO 15216-1 2017, using Mengovirus as a process control. The SARS-CoV-2, RVA, NoV, HEV and Mengovirus real-time RT-PCRs were applied to extracted RNA samples. Moreover, the conventional multiplex RT-PCR assay for RVA VP7/VP4 genotyping and Sanger sequencing was employed. According to the results, the RVA is widespread in the Croatian environment as it was detected in 52% of surface/sewage samples and 22.7% of bivalves. RVA genotyping revealed the presence of multiple G and P genotypes, with the notable finding of the emerging human G3 equine-like strains. The ongoing research will extend our knowledge on SARS-CoV-2, NoV and HEV presence in these samples, with the results expected to be presented at the conference. The continuous monitoring of potential viral threats in the environment within the One health approach, can provide valuable information on circulating viral threats, as an early indicator of their presence in humans.

SARS-CoV-2, rotavirus A, norovirus, hepatitis E virus, environment, wastewater, surface water, bivalve molluscan shellfish

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