SpringerLink
Log in

Interactions between Salmonella typhimurium and Acanthamoeba polyphaga, and Observation of a New Mode of Intracellular Growth within Contractile Vacuoles

  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

Acanthamoeba polyphaga feeding on Salmonella typhimurium in a simple model biofilm were observed by light microscopy and a detailed record of interactions kept by digital image capture and image analysis. A strain of S. typhimurium SL1344 carrying a fis:gfp reporter construct (pPDT105) was used to assess intracellular growth in A. polyphaga on non-nutrient agar (NNA) plates. Invasion of the contractile vacuole (CV) was observed at a frequency of 1:100–1000 acanthamoebae at 35°C. The salmonellae contained in CVs illustrated significant up-regulation of fis relative to extracellular bacteria, indicating that they were in the early stages of logarithmic growth, and reached numbers of 100–200 cells per vacuole after 4 days. This is the first report of this mode of intracellular growth. Up-regulation of fis was also observed in a proportion of S. typhimurium cells contained within food vacuoles. Filamentation of S. typhimurium and E. coli cells was frequently observed in coculture with A. polyphaga on NNA plates, with bacterial cells reaching lengths of up to 500 µm after 10 days’ incubation at 35°C. A. polyphaga was also seen to mediate bacterial translocation over the agar surface; egested salmonellae subsequently formed microcolonies along amoebal tracks. This illustrated intracellular survival of a fraction of the S. typhimurium population. These phenomena suggest that protozoa such as A. polyhaga may play an important role in the ecology of S. typhimurium in soil and aquatic environments.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. RD Allen (2000) ArticleTitleThe contractile vacuole and its membrane dynamics. BioEssays 22 1035–1042 Occurrence Handle10.1002/1521-1878(200011)22:11<1035::AID-BIES10>3.3.CO;2-1 Occurrence Handle1:CAS:528:DC%2BD3MXmslShs7w%3D Occurrence Handle11056480

    Article  CAS  PubMed  Google Scholar 

  2. J Barker TJ Humphrey MWR Brown (1999) ArticleTitleSurvival of Escherichia coli 0157 in a soil protozoan: implications for disease. FEMS Microbiol Lett 173 291–295 Occurrence Handle10.1016/S0378-1097(99)00076-2 Occurrence Handle1:CAS:528:DyaK1M**tFWrt7k%3D Occurrence Handle10227158

    Article  CAS  PubMed  Google Scholar 

  3. M Bianchi (1989) ArticleTitleUnusual bloom of star-like prosthecate bacteria and filaments as a consequence of grazing pressure. Microb Ecol 17 137–141

    Google Scholar 

  4. G Brandi L Salvaggio F Cattabeni O Cantoni (1991) ArticleTitleCytocidal and filamentation response of Escherichia coli cells exposed to low concentrations of hydrogen peroxide and hydroxyl radical scavengers. Environ Mol Mutagen 18 22–27 Occurrence Handle1:CAS:528:DyaK3MXlt12mur0%3D Occurrence Handle1907540

    CAS  PubMed  Google Scholar 

  5. A Cegielska A Taylor (1985) ArticleTitleThe sfiA 11 mutation prevents filamentation in response to cell wall damage only in a recA + genetic background. Mol Gen Genet 201 537–542 Occurrence Handle1:CAS:528:DyaL28Xot1ygtA%3D%3D Occurrence Handle3911030

    CAS  PubMed  Google Scholar 

  6. JD Cirillo S Falkow LS Tompkins LE Bermudez (1997) ArticleTitleInteraction of Mycobacterium avium with environmental amoebae enhances virulence. Infect Immun 65 3759–3767 Occurrence Handle1:CAS:528:DyaK2sXlvFKnsbo%3D Occurrence Handle9284149

    CAS  PubMed  Google Scholar 

  7. JD Cirillo SLG Cirillo L Yan LE Bermudez S Falkow LS Tompkins (1999) ArticleTitleIntracellular growth in Acanthamoeba castellanii affects monocyte entry mechanisms and enhances virulence of Legionella pneumophila. Infect Immun 67 4427–4434 Occurrence Handle1:CAS:528:DyaK1MXlsFCmt74%3D Occurrence Handle10456883

    CAS  PubMed  Google Scholar 

  8. W Drozanski (1978) ArticleTitleActivity and distribution of bacteriolytic N-acetyl-muramidase during growth of Acanthamoeba castelanii in axenic culture. Acta Microbiol Pol 28 123–133

    Google Scholar 

  9. A Essig M Heinemann U Simnacher R Marre (1997) ArticleTitleInfection of Acanthamoeba castellanii by Chlamydia pneumoniae. Appl Environ Microbiol 63 1396–1399 Occurrence Handle1:CAS:528:DyaK2s**sVWmsbg%3D Occurrence Handle9097437

    CAS  PubMed  Google Scholar 

  10. F Ekelund R Ronn (1994) ArticleTitleNotes on protozoa in agricultural soil with emphasis on heterotrophic flagellates and naked amoebae and their ecology. FEMS Microbiol Rev 15 321–353 Occurrence Handle10.1016/0168-6445(94)90068-X Occurrence Handle1:CAS:528:DyaK2M**vVKlt78%3D Occurrence Handle7848658

    Article  CAS  PubMed  Google Scholar 

  11. AM Goulsbra C Edwards MP Gallagher (2001) ArticleTitleSurface hygiene monitored using a reporter of fis expression in Escherichia coli. J Appl Microbiol 91 104–109 Occurrence Handle10.1046/j.1365-2672.2001.01344.x Occurrence Handle1:CAS:528:DC%2BD3MXlslaltL0%3D Occurrence Handle11442719

    Article  CAS  PubMed  Google Scholar 

  12. H Gude (1979) ArticleTitleGrazing by protozoa as selection factor for activated sludge bacteria. Microb Ecol 5 225–237

    Google Scholar 

  13. RM Harshey T Matsuyama (1994) ArticleTitleDimorphic transition in Escherichia coli and Salmonella typhimurium: surface-induced differentiation into hyperflagellate swarmer cells. Proc Natl Acad Sci USA 91 8631–8635 Occurrence Handle1:CAS:528:DyaK2cXlslCqtr4%3D Occurrence Handle8078935

    CAS  PubMed  Google Scholar 

  14. J Heuser Q Zhu M Clarke (1993) ArticleTitleProton pumps populate the contractile vacuoles of Dictyostelium amoebae. J Cell Biol 121 1311–1327 Occurrence Handle1:CAS:528:DyaK3s**sFWit78%3D Occurrence Handle8509452

    CAS  PubMed  Google Scholar 

  15. SK Hoiseth BA Stocker (1981) ArticleTitleAromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature 291 238–239 Occurrence Handle1:CAS:528:DyaL3MXkvFeltLg%3D Occurrence Handle7015147

    CAS  PubMed  Google Scholar 

  16. CH King EB Shotts Jr RE Wooley KG Porter (1988) ArticleTitleSurvival of coliforms and bacterial pathogens within protozoa during chlorination. Appl Environ Microbiol 54 3023–3033 Occurrence Handle1:CAS:528:DyaL1MXlsVekuw%3D%3D Occurrence Handle3223766

    CAS  PubMed  Google Scholar 

  17. WJ Kwan (Eds) (1973) The Biology of Amoeba. Academic Press New York

    Google Scholar 

  18. TMC Ly HE Muller (1990) ArticleTitleIngested Listeria monocytogenes survive and multiply in protozoa. J Med Microbiol 33 51–54 Occurrence Handle1:STN:280:By6D3sbisVQ%3D Occurrence Handle2121990

    CAS  PubMed  Google Scholar 

  19. CL Marolda B Hauroder R Michel MA Valvano (1999) ArticleTitleIntracellular survival and saprophytic growth of isolates from the Burkholderia cepacia complex of free-living amoebae. Microbiology 145 1509–1517 Occurrence Handle1:CAS:528:DyaK1MXkvFaht7o%3D Occurrence Handle10439391

    CAS  PubMed  Google Scholar 

  20. R Osuna D Lienau KT Hughes RC Johnson (1995) ArticleTitleSequence, regulation, and functions of fis in Salmonella typhimurium. J Bacteriol 177 2021–2032 Occurrence Handle1:CAS:528:DyaK2MXkvFelsrc%3D Occurrence Handle7536730

    CAS  PubMed  Google Scholar 

  21. S Rodríguez-Zaragosa (1994) ArticleTitleEcology of free-living amoebae. Crit Rev Microbiol 20 225–241 Occurrence Handle1:STN:280:ByqC3c7htlw%3D Occurrence Handle7802958

    CAS  PubMed  Google Scholar 

  22. TJ Rowbotham (1980) ArticleTitlePreliminary report on the pathogenicity of Legionella pneumophila for freshwater and soil amoebae. J Clin Pathol 33 1179–1183 Occurrence Handle1:STN:280:Bi6D1Mvks1A%3D Occurrence Handle7451664

    CAS  PubMed  Google Scholar 

  23. W Schlimme M Marchiani K Hanselmann B Jenni (1997) ArticleTitleGene transfer between bacteria within digestive vacuoles of protozoa. FEMS Microb Ecol 23 239–247 Occurrence Handle10.1016/S0168-6496(97)00028-7 Occurrence Handle1:CAS:528:DyaK2sXlt1Ogsbo%3D

    Article  CAS  Google Scholar 

  24. H Sesaki EFS Wong C Siu (1997) ArticleTitleThe cell adhesion molecule DdCAD-1 in Dictyostelium is targeted to the cell surface by a nonclassical transport pathway involving contractile vacuoles. J Cell Biol 138 939–951 Occurrence Handle10.1083/jcb.138.4.939 Occurrence Handle1:CAS:528:DyaK2sXlsleiu78%3D Occurrence Handle9265658

    Article  CAS  PubMed  Google Scholar 

  25. S Shikano LS Luckinbill Y Kurihara (1990) ArticleTitleChanges of traits in a bacterial population associated with protozoal predation. Microb Ecol 20 75–84

    Google Scholar 

  26. M Steinert L Emody R Amann J Hacker (1997) ArticleTitleResuscitation of viable but nonculturable Legionella pneumophila Philadelphia JR32 by Acanthamoeba castellanii. Appl Environ Microbiol 63 2047–2053 Occurrence Handle1:CAS:528:DyaK2sXjtFGlsr4%3D Occurrence Handle9143134

    CAS  PubMed  Google Scholar 

  27. J Xu RC Johnson (1995) ArticleTitleIdentification of genes negatively regulated by Fis: Fis and RpoS comodulate growth-phase-dependent gene expression in Escherichia coli. J Bacteriol 177 938–947 Occurrence Handle1:CAS:528:DyaK2MXjs1Khtbs%3D Occurrence Handle7860604

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by BBSRC grant 88/MM109738.

Author information

Authors and Affiliations

  1. Department of Biological Science, University of Warwick, Coventry, CV4 7AL, UK

    W. H. Gaze & E. M. H. Wellington

  2. Institute of Mathematics, University of Warwick, Coventry, CV4 7AL, UK

    N. Burroughs

  3. Institute of Cell and Molecular Biology, Biology Division, University of Edinburgh, Mayfield Road, Edinburgh EH9 3JR, UK

    M. P. Gallagher

Authors
  1. W. H. Gaze
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Burroughs
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. P. Gallagher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E. M. H. Wellington
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to W. H. Gaze.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gaze, W., Burroughs, N., Gallagher, M. et al. Interactions between Salmonella typhimurium and Acanthamoeba polyphaga, and Observation of a New Mode of Intracellular Growth within Contractile Vacuoles . Microb Ecol 46, 358–369 (2003). https://doi.org/10.1007/s00248-003-1001-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-003-1001-3

Keywords

Search

Navigation