Porphyromonas gingivalis and Prevotella intermedia are obligate anaerobe gram-negative periodontopathogenic bacteria. Free-living amoebae, such as Acanthamoeba spp., are well known as environmental hosts of several human pathogens, such as Franciscella tularensis, Chlamydia pneumoniae, Legionella pneumophila and Mycobacteria spp. This study tested the ability of P. gingivalis and P. intermedia to become internalized, to survive and replicate in Acanthamoeba castellani. Our results show for the first time that P. gingivalis and P. intermedia isolated from periodontitis patients are capable of infecting A. castellani cells in vitro and are able to survive and multiply intracellularly. From our experimental data it can be suggested that periodontopathogenic bacteria might be conditioned in the evolution by amoebae and free-living amoebae can act as an environmental reservoir for these pathogens.