Shigella species and Acanthamoeba castellanii share the same ecological niches, and their interaction has been addressed in a limited number of research. However, there are still uncertain aspects and discrepant findings of this interaction. In the present study, the effects of the bacterial growth phase, cocultivation temperature and the type of culture media on the interaction of A. castellanii with Shigella dysenteriae, Shigella sonnei and Shigella flexneri were evaluated. In nutrient‐poor page's amoeba saline (PAS) medium, the number of recovered bacteria and the uptake rates were significantly higher in stationary phase cells than logarithmic phase cells. However, no significant differences were observed in the number of recovered bacteria and the uptake rates between logarithmic and stationary phase cells in nutrient‐rich peptone–yeast extract–glucose (PYG) medium. While the number of recovered bacteria was significantly higher in nutrient‐rich than nutrient‐poor media, in all the three Shigella species, the bacterial uptake rates were significantly higher in nutrient‐poor than nutrient‐rich media at both cocultivation temperatures. In both nutrient‐poor and nutrient‐rich media and at both cocultivation temperatures, the number of viable Shigella species after 24 h incubation were not influenced by the presence of A. castellanii. Although Shigella species did not proliferate in A. castellanii trophozoites, a considerable number of bacteria were survived in the trophozoites up to 15 days. From the public health perspective, the results of this study are important for further understanding of the nature of the interaction of these organisms and to deal with Shigella species in the environment.
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