Humans are frequently exposed to Listeria monocytogenes, and high numbers may be ingested during consumption of certain types of food. However, epidemiological investigations show that listeriosis is a rare disease. Risk assessment studies using an animal mouse model indicate that almost all L. monocytogenes serovars present in food have clear virulent properties. The intravenous dose causing infection in 50% (IV ID50) of mice not previously exposed to L. monocytogenes (nonprotected mice) was 1.8 log10 units. For mice previously exposed to L. monocytogenes (immunologically protected mice), the IV ID50 was 5.6 log10 units. The ID50 of orally exposed nonprotected mice amounted to 6.5 log10 units, and no significant effects of type of food (water/milk) and storage time at 5°C (milk) were observed. The oral ID50 of immunologically protected mice was >9.0 log10 units. Furthermore, there was approximately 1–2 log10 difference between the ID50 and the lethal dose causing death in 50% (LD50). The results show that both the intestinal barrier and the specific immune defense mechanism are highly effective in preventing infection of mice orally exposed to L. monocytogenes. Delaying the immune defense had no effect on the protective activity of the intestinal barrier, indicating that these protective mechanisms operate independently. The risk assessment results obtained in the mouse model support the epidemiological finding that listeriosis is a rare disease in humans, despite frequent exposure to the organism.