The significance of multiplicity of infection (moi) for invasiveness and intracellular multiplication of in Caco-2 and HeLa cell monolayers was investigated. A low moi (1:1) resulted in recovery of significantly more when these bacteria were used to infect either cell line. At high moi (100:1), the percentage recovery of bacteria was comparatively low, even after extensive invasion and intracellular multiplication. Microscopic analysis of Giemsa- and immunofluorescent-stained infected monolayers revealed extensive cell disruption and exposure of the internalised bacteria to the bactericidal effect of gentamicin. By contrast, a low moi resulted in minimal cytopathic effects and evidence of cell to cell spread by was consistently observed in HeLa and J774, but not in Caco-2 cell lines. Nevertheless, the use of HeLa and Caco-2 cell monolayers enabled a clear distinction to be made between invasive () and non-invasive spp. ( and LLO19). The use of a low moi with HeLa cell monolayers provided a reliable tissue-culture model of infection for


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