Skip to main content
SpringerLink
Log in

Assessing Pseudomonas aeruginosa Virulence Using a Nonmammalian Host: Dictyostelium discoideum

  • Protocol
  • First Online:
Pseudomonas Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1149))

Abstract

Dictyostelium discoideum, a soil amoeba, can be used as an alternative host to study the virulence of various bacterial species, including Pseudomonas aeruginosa. A simple quantitative test based on the ability of D. discoideum to grow on a bacterial lawn has been developed using this amoeba to assay the virulence of P. aeruginosa strains. The assay needs to be customized for the strains to be tested in order to be able to discriminate between virulent and avirulent P. aeruginosa strains. These steps are described in this protocol.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 299.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Froquet R, Lelong E, Marchetti A, Cosson P (2009) Dictyostelium discoideum: a model host to measure bacterial virulence. Nat Protoc 4:25–30

    Article  CAS  Google Scholar 

  2. Vilches S, Jimenez N, Merino S, Tomas JM (2011) The Aeromonas dsbA mutation decreased their virulence by triggering type III secretion system but not flagella production. Microb Pathog 52:130–139

    Article  Google Scholar 

  3. Daher RK, Filion G, Tan SG, Dallaire-Dufresne S, Paquet VE, Charette SJ (2011) Alteration of virulence factors and rearrangement of pAsa5 plasmid caused by the growth of Aeromonas salmonicida in stressful conditions. Vet Microbiol 152:353–360

    Article  CAS  Google Scholar 

  4. Froquet R, Cherix N, Burr SE, Frey J, Vilches S, Tomas JM, Cosson P (2007) Alternative host model to evaluate Aeromonas virulence. Appl Environ Microbiol 73:5657–5659

    Article  CAS  Google Scholar 

  5. Benghezal M, Fauvarque MO, Tournebize R, Froquet R, Marchetti A, Bergeret E, Lardy B, Klein G, Sansonetti P, Charette SJ, Cosson P (2006) Specific host genes required for the killing of Klebsiella bacteria by phagocytes. Cell Microbiol 8:139–148

    Article  CAS  Google Scholar 

  6. Pan YJ, Lin TL, Hsu CR, Wang JT (2011) Use of a Dictyostelium model for isolation of genetic loci associated with phagocytosis and virulence in Klebsiella pneumoniae. Infect Immun 79:997–1006

    Article  CAS  Google Scholar 

  7. Hagele S, Kohler R, Merkert H, Schleicher M, Hacker J, Steinert M (2000) Dictyostelium discoideum: a new host model system for intracellular pathogens of the genus Legionella. Cell Microbiol 2:165–171

    Article  CAS  Google Scholar 

  8. Solomon JM, Rupper A, Cardelli JA, Isberg RR (2000) Intracellular growth of Legionella pneumophila in Dictyostelium discoideum, a system for genetic analysis of host-pathogen interactions. Infect Immun 68:2939–2947

    Article  CAS  Google Scholar 

  9. Lu H, Clarke M (2005) Dynamic properties of Legionella-containing phagosomes in Dictyostelium amoebae. Cell Microbiol 7:995–1007

    Article  CAS  Google Scholar 

  10. Shevchuk O, Steinert M (2009) Screening of virulence traits in Legionella pneumophila and analysis of the host susceptibility to infection by using the Dictyostelium host model system. Methods Mol Biol 470:47–56

    Article  CAS  Google Scholar 

  11. Skriwan C, Fajardo M, Hagele S, Horn M, Wagner M, Michel R, Krohne G, Schleicher M, Hacker J, Steinert M (2002) Various bacterial pathogens and symbionts infect the amoeba Dictyostelium discoideum. Int J Med Microbiol 291:615–624

    Article  Google Scholar 

  12. Peracino B, Wagner C, Balest A, Balbo A, Pergolizzi B, Noegel AA, Steinert M, Bozzaro S (2006) Function and mechanism of action of Dictyostelium Nramp1 (Slc11a1) in bacterial infection. Traffic 7:22–38

    Article  CAS  Google Scholar 

  13. Hagedorn M, Soldati T (2007) Flotillin and RacH modulate the intracellular immunity of Dictyostelium to Mycobacterium marinum infection. Cell Microbiol 9:2716–2733

    Article  CAS  Google Scholar 

  14. Solomon JM, Leung GS, Isberg RR (2003) Intracellular replication of Mycobacterium marinum within Dictyostelium discoideum: efficient replication in the absence of host coronin. Infect Immun 71:3578–3586

    Article  CAS  Google Scholar 

  15. Hagedorn M, Rohde KH, Russell DG, Soldati T (2009) Infection by tubercular mycobacteria is spread by nonlytic ejection from their amoeba hosts. Science 323:1729–1733

    Article  CAS  Google Scholar 

  16. Vial L, Groleau MC, Lamarche MG, Filion G, Castonguay-Vanier J, Dekimpe V, Daigle F, Charette SJ, Deziel E (2010) Phase variation has a role in Burkholderia ambifaria niche adaptation. ISME J 4:49–60

    Article  Google Scholar 

  17. Aubert DF, Flannagan RS, Valvano MA (2008) A novel sensor kinase-response regulator hybrid controls biofilm formation and type VI secretion system activity in Burkholderia cenocepacia. Infect Immun 76:1979–1991

    Article  CAS  Google Scholar 

  18. Hasselbring BM, Patel MK, Schell MA (2011) Dictyostelium discoideum as a model system for identification of Burkholderia pseudomallei virulence factors. Infect Immun 79:2079–2088

    Article  CAS  Google Scholar 

  19. Bonifait L, Charette SJ, Filion G, Gottschalk M, Grenier D (2011) Amoeba host model for evaluation of Streptococcus suis virulence. Appl Environ Microbiol 77:6271–6273

    Article  CAS  Google Scholar 

  20. Cosson P, Zulianello L, Join-Lambert O, Faurisson F, Gebbie L, Benghezal M, Van Delden C, Curty LK, Kohler T (2002) Pseudomonas aeruginosa virulence analyzed in a Dictyostelium discoideum host system. J Bacteriol 184:3027–3033

    Article  CAS  Google Scholar 

  21. Pukatzki S, Kessin RH, Mekalanos JJ (2002) The human pathogen Pseudomonas aeruginosa utilizes conserved virulence pathways to infect the social amoeba Dictyostelium discoideum. Proc Natl Acad Sci U S A 99:3159–3164

    Article  CAS  Google Scholar 

  22. Alibaud L, Kohler T, Coudray A, Prigent-Combaret C, Bergeret E, Perrin J, Benghezal M, Reimmann C, Gauthier Y, van Delden C, Attree I, Fauvarque MO, Cosson P (2007) Pseudomonas aeruginosa virulence genes identified in a Dictyostelium host model. Cell Microbiol 10:729–740

    Article  Google Scholar 

  23. Lima WC, Lelong E, Cosson P (2011) What can Dictyostelium bring to the study of Pseudomonas infections? Semin Cell Dev Biol 22:77–81

    Article  CAS  Google Scholar 

  24. Lelong E, Marchetti A, Simon M, Burns JL, van Delden C, Kohler T, Cosson P (2010) Evolution of Pseudomonas aeruginosa virulence in infected patients revealed in a Dictyostelium discoideum host model. Clin Microbiol Infect 17:1415–1420

    Article  Google Scholar 

  25. Sperandio D, Decoin V, Latour X, Mijouin L, Hillion M, Feuilloley MG, Orange N, Merieau A (2012) Virulence of the Pseudomonas fluorescens clinical strain MFN1032 towards Dictyostelium discoideum and macrophages in relation with type III secretion system. BMC Microbiol 12:223

    Article  CAS  Google Scholar 

  26. Vallet-Gely I, Novikov A, Augusto L, Liehl P, Bolbach G, Pechy-Tarr M, Cosson P, Keel C, Caroff M, Lemaitre B (2010) Association of hemolytic activity of Pseudomonas entomophila, a versatile soil bacterium, with cyclic lipopeptide production. Appl Environ Microbiol 76:910–921

    Article  CAS  Google Scholar 

  27. Janjua HA, Segata N, Bernabo P, Tamburini S, Ellen A, Jousson O (2012) Clinical populations of Pseudomonas aeruginosa isolated from acute infections show a wide virulence range partially correlated with population structure and virulence gene expression. Microbiology 158:2089–2098

    Article  CAS  Google Scholar 

  28. Bradbury RS, Reid DW, Inglis TJ, Champion AC (2011) Decreased virulence of cystic fibrosis Pseudomonas aeruginosa in Dictyostelium discoideum. Microbiol Immunol 55:224–230

    Article  CAS  Google Scholar 

  29. Kukavica-Ibrulj I, Bragonzi A, Paroni M, Winstanley C, Sanschagrin F, O'Toole GA, Levesque RC (2008) In vivo growth of Pseudomonas aeruginosa strains PAO1 and PA14 and the hypervirulent strain LESB58 in a rat model of chronic lung infection. J Bacteriol 190:2804–2813

    Article  CAS  Google Scholar 

  30. Klockgether J, Munder A, Neugebauer J, Davenport CF, Stanke F, Larbig KD, Heeb S, Schock U, Pohl TM, Wiehlmann L, Tummler B (2010) Genome diversity of Pseudomonas aeruginosa PAO1 laboratory strains. J Bacteriol 192:1113–1121

    Article  CAS  Google Scholar 

  31. Fey P, Kowal AS, Gaudet P, Pilcher KE, Chisholm RL (2007) Protocols for growth and development of Dictyostelium discoideum. Nat Protoc 2:1307–1316

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Katherine H. Tanaka and Luc Trudel for critical reading of the manuscript. We are grateful to Sok Gheck E. Tan for her assistance with the protocol. This work was funded by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada (NSERC), a Fonds de la Recherche du Québec en Santé (FRQS) grant, and a Young Researcher grant from the FRQS Réseau en santé respiratoire (RSR). S.J.C. is a research scholar of the FRQS.

Author information

Authors and Affiliations

  1. Institut de Biologie Intégrative et des Systèmes (IBIS), Pavillon Charles-Eugène-Marchand, Université Laval, Quebec, QC, Canada

    Geneviève Filion & Steve J. Charette

  2. Centre de recherche de l’Institut Universitaire de Cardiologie et de Pneumologie de Québec (CRIUCPQ), Quebec, QC, Canada

    Geneviève Filion & Steve J. Charette

  3. Département de biochimie, de microbiologie et de bio-informatique, Faculté de sciences et de génie, Université Laval, Quebec, QC, Canada

    Steve J. Charette

Authors
  1. Geneviève Filion
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Steve J. Charette
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Steve J. Charette .

Editor information

Editors and Affiliations

  1. MRC Centre for Molecular Bacteriology and Infection (CMBI) Department of Life Sciences, Imperial College, London, UK

    Alain Filloux

  2. Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Granada, Spain

    Juan-Luis Ramos

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Filion, G., Charette, S.J. (2014). Assessing Pseudomonas aeruginosa Virulence Using a Nonmammalian Host: Dictyostelium discoideum . In: Filloux, A., Ramos, JL. (eds) Pseudomonas Methods and Protocols. Methods in Molecular Biology, vol 1149. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-0473-0_51

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0473-0_51

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-0472-3

  • Online ISBN: 978-1-4939-0473-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation