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Applied and Environmental Microbiology, June 2004, p. 3706-3714, Vol. 70, No. 6
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.6.3706-3714.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Uptake and Replication of Salmonella enterica in Acanthamoeba rhysodes

Dilek Tezcan-Merdol,1 Marianne Ljungström,2 Jadwiga Winiecka-Krusnell,2 Ewert Linder,1,2 Lars Engstrand,1,2 and Mikael Rhen1,2*

Microbiology and Tumor Biology Center, Karolinska Institute, 171 77 Stockholm,1 Swedish Institute for Infectious Disease Control, 171 82 Solna, Sweden2

Received 19 September 2003/ Accepted 1 February 2004

The ability of salmonellae to become internalized and to survive and replicate in amoebae was evaluated by using three separate serovars of Salmonella enterica and five different isolates of axenic Acanthamoeba spp. In gentamicin protection assays, Salmonella enterica serovar Dublin was internalized more efficiently than Salmonella enterica serovar Enteritidis or Salmonella enterica serovar Typhimurium in all of the amoeba isolates tested. The bacteria appeared to be most efficiently internalized by Acanthamoeba rhysodes. Variations in bacterial growth conditions affected internalization efficiency, but this effect was not altered by inactivation of hilA, a key regulator in the expression of the invasion-associated Salmonella pathogenicity island 1. Microscopy of infected A. rhysodes revealed that S. enterica resided within vacuoles. Prolonged incubation resulted in a loss of intracellular bacteria associated with morphological changes and loss of amoebae. In part, these alterations were associated with hilA and the Salmonella virulence plasmid. The data show that Acanthamoeba spp. can differentiate between different serovars of salmonellae and that internalization is associated with cytotoxic effects mediated by defined Salmonella virulence loci.

* Corresponding author. Mailing address: Microbiology and Tumor Biology Center, Karolinska Institute, Nobels Väg 16, 171 77 Stockholm, Sweden. Phone: 46 8 728 3794. Fax: 46 8 330498. E-mail: mikael.rhen{at}mtc.ki.se.

Applied and Environmental Microbiology, June 2004, p. 3706-3714, Vol. 70, No. 6
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.6.3706-3714.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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