Antiviral Effects of Mulberry (Morus Alba) Juice and Its Fractions on Foodborne Viral Surrogates

Norovirus infection is a major cause of nonbacterial foodborne outbreaks worldwide, but no specific treatments are available yet. In this study, we investigated the antiviral activity of mulberry (Morus alba, Ma) juice and its fractions on murine norovirus-1 (MNV-1) and feline calicivirus-F9 (FCV-F9) as human norovirus surrogates using cytopathic effect inhibition, plaque reduction, and RNA expression assays. In time-of-addition experiments, Ma juice was found to be effective in reducing the infectivity of MNV-1 and FCV-F9 in the pre- and co-treatments. The effective concentration for 50% reduction was approximately 0.005% juice (relative to 100% natural juice) and 0.25% juice for MNV-1 and FCV-F9, respectively. Ma juice at 0.1% exhibited about 60% reduction of the MNV-1 polymerase gene expression, confirming the inhibition of viral replication. In an attempt to identify active components with antiviral activities, Ma-F1 (<3 kDa) and Ma-F2 (>3 kDa) were examined to show that Ma-F2 was more effective than Ma-F1 in all modes, except for pre-virus treatment. Nevertheless, two major polyphenolic compounds of Ma juice, cyanidin-3-glucoside and cyanidin-3-rutinoside, showed antiviral activity in the co-treatment mode. Our results suggest that Ma juice and its fractions may inhibit internalization and replication of MNV-1, whereas it may influence adherence or internalization of FCV-F9 virions. Ma juice may prove useful in the prevention of foodborne viral infection.