Helicobacter pylori is presumed to infect gastric tissue via the oral cavity in childhood, whereas risk factors for H. pylori infection in the oral cavity are unknown. In this study, we analysed the effects of Streptococcus mutans, a major cariogenic bacterial species, on H. pylori colonisation in the oral cavity, as well as gastric tissue. Rats in the weaning period were infected with S. mutans in the oral cavity, then fed a caries-inducing diet to facilitate S. mutans colonisation. One month after S. mutans infection, rats were infected with H. pylori in the oral cavity; rats were then euthanised at 1 month after H. pylori infection. H. pylori was detected in the oral cavities of rats infected with both S. mutans and H. pylori, but not in rats infected with H. pylori alone. In addition, H. pylori colonisation in the gastric tissue and typical gastrointestinal damage were observed in rats infected with both S. mutans and H. pylori. When H. pylori was co-cultured with in vitro biofilm formed by S. mutans, a large number of H. pylori bacteria invaded the biofilm formed by S. mutans. Our results suggest that S. mutans is involved in the establishment of H. pylori infection. Helicobacter pylori, a helix-shaped gram-negative microaerophilic bacterium, is a major causative agent of gastric cancer and gastric ulcers 1. More than half of the world's population is infected with H. pylori 2 , which is presumably acquired mainly via the oral cavity in childhood 3,4. Molecular biological techniques have reportedly revealed H. pylori in oral specimens 5-7. The presence of H. pylori in the oral cavity has been related to the detection of H. pylori in the gastric tissue 8. However, details regarding risk factors of H. pylori infection in the oral cavity have not been clarified, which may explain the current difficulty in elimination of H. pylori infection. Streptococcus mutans, a gram-positive facultative anaerobe, is a major causative pathogen of dental caries 9. S. mutans is acquired in the oral cavity during early childhood, mainly via mother-to-child transmission 10. The aetiology of dental caries caused by S. mutans was clarified in the early 1960s 11 ; S. mutans metabolises sucrose to form a biofilm on the tooth surface, followed by demineralisation of the tooth. Nevertheless, eradication of S. mutans from the oral cavity and dental caries remains difficult 12. Some epidemiological studies have revealed that patients with dental caries or poor oral hygiene were more likely to harbour H. pylori in oral cavity or gastric tissue 13,14. These findings suggest that the presence of cariogenic bacteria is involved in infection of the oral cavity with H. pylori. To the best of our knowledge, no clear evidence has been obtained regarding the effects of S. mutans on H. pylori infection in an animal model. In the present study, we hypothesised that S. mutans colonisation in the oral cavity may be involved in H. pylori colonisation in both oral cavity and gastric tissue. Therefore, we constructed a rat co-infection model with S. mut...