High persistence of hepatitis A and E viruses during simulated gastrointestinal digestion across diverse food matrices

Hepatitis A virus (HAV) and hepatitis E virus (HEV) are major causes of foodborne viral infections worldwide. Although both viruses are known to be transmitted via the fecal-oral route, their persistence during passage through the human gastrointestinal tract, particularly in the presence of different food matrices, remains poorly characterized. In this study, the persistence of HAV and HEV was evaluated using two complementary in vitro digestion models, the static INFOGEST 2.0 protocol and the dynamic TIM-1 system, simulating conditions of the stomach and small intestine. Different food matrices associated with viral transmission, including berries, oysters, lettuce (HAV) and pork liver pâté (HEV), were investigated. HAV infectivity was assessed by plaque assays, while HAV and HEV genome were quantified by RT-qPCR following PMAxx pretreatment. For both viruses, viral levels at the end of the digestion were comparable to initial concentration across all tested matrices and in both models. Although minor variations were observed at specific time points, these were not statistically significant. Overall, the results indicate that HAV and HEV exhibit high persistence under simulated gastrointestinal conditions, with limited influence of the food matrix. These findings suggest that intrinsic viral resistance rather than matrix-dependent protection governs their survival during digestion, highlighting a potential risk to consumers even at low contamination levels. This study underscores the need to better account for viral persistence during digestion in food safety risk assessments and control strategies.