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The amoeba  Dictyostelium discoideum  has been used as a model organism to study host-pathogen interaction in many intracellular bacteria.  Francisella tularensis  is a Gram-negative, highly infectious bacterium that causes the zoonotic disease tularemia. The bacterium is able to replicate in different phagocytic and nonphagocytic cells including mammalian, amoebae, and arthropod cells. The aim of this study was to determine the optimal temperature and infection dose in the interaction of  Francisella novicida  with  D. discoideum  in order to establish a model of  Francisella  infection in the social amoeba. The amoeba cells were infected with a different multiplicity of infection (5, 10, and 100) and incubated at different temperatures (22, 25, 27, 30, and 37°C). The number of intracellular bacteria within  D. discoideum , as well as cytotoxicity, was determined at 2, 4, 24, 48, and 72 hours after infection. Our results showed that the optimal temperature for  Francisella  intracellular replication within amoeba is 30°C with the MOI of 10. We can conclude that this MOI and temperature induced the optimal growth of bacteria in  Dictyostelium  with low cytotoxicity.

Optimisation of External Factors for the Growth of Francisella novicida within Dictyostelium discoideum