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Background   Acanthamoeba  can interact with a wide range of microorganisms such as viruses, algae, yeasts, protists and bacteria including  Legionella pneumophila ,  Pseudomonas aeruginosa ,  Vibrio cholerae ,  Helicobacter pylori ,  Listeria monocytogenes ,  Mycobacterium  spp., and  Escherichia coli . In this capacity,  Acanthamoeba  has been suggested as a vector in the transmission of bacterial pathogens to the susceptible hosts.    Methods  Here, we used a keratitis isolate of  A. castellanii  of the T4 genotype and studied its interactions with two bacterial genera which have not been tested before,  Enterobacter aerogenes , and  Aeromonas hydrophila , as well as  E. coli . Assays were performed to determine bacterial association with and invasion of  A. castellanii . Additionally, bacterial survival intracellular of  A. castellanii  trophozoites as well as cysts was determined.    Results  All three bacterial isolates tested, associated, invaded, and survived inside  A. castellanii  trophozoites as well as  A. castellanii  cysts. However,  E. aerogenes  and  E. coli  exhibited significantly reduced association with and invasion of  A. castellanii  as compared with  A. hydrophila  ( P  < 0.01 using paired T-test, one tail distribution). In the long term survival assays, all three bacterial isolates tested remained viable inside  A. castellanii  trophozoites, while amoeba remained intact; however  A. hydrophila  exhibited higher survival inside amoebae (14.54 ± 3.3 bacteria:amoeba ratio) compared with  E. aerogenes  (3.96 ± 0.7 bacteria:amoeba ratio) and  E. coli  (5.85 ± 1.1 bacteria:amoeba ratio).  A. hydrophila ,  E. coli , and  E. aerogenes  remained viable during the encystment process and exhibited higher levels of recovery from mature cysts (14.13 ± 0.89  A. hydrophila :amoeba ratio, 10.13 ± 1.17  E. aerogenes :amoeba ratio, and 11.95 ± 0.7  E. coli :amoeba ratio).    Conclusions   A. hydrophila  and  E. aerogenes  also joined the ranks of other bacteria that could benefit from  A. castellanii . Because cysts can be airborne, these findings suggest that  Acanthamoeba  is a potential vector in the transmission of  A. hydrophila  and  E. aerogenes  to susceptible hosts.

Acanthamoeba castellanii of the T4 genotype is a potential environmental host for Enterobacter aerogenes and Aeromonas hydrophila