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Impact of Non-Legionella Bacteria on the Uptake and Intracellular Replication of Legionella pneumophila in Acanthamoeba castellanii and Naegleria lovaniensis

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Abstract

In aquatic environments, Legionella pneumophila survives, in association with other bacteria, within biofilms by multiplying in free-living amoebae. The precise mechanisms underlying several aspects of the uptake and intracellular replication of L. pneumophila in amoebae, especially in the presence of other bacteria, remain unknown. In the present study, we examined the competitive effect of selected non-Legionella bacteria (Escherichia coli, Aeromonas hydrophila, Flavobacterium breve, and Pseudomonas aeruginosa) on the uptake of L. pneumophila serogroup 1 by the amoebae Acanthamoeba castellanii and Naegleria lovaniensis. We also investigated their possible influence on the intracellular replication of L. pneumophila in both amoeba species. Our results showed that the non-Legionella bacteria did not compete with L. pneumophila for uptake, suggesting that the amoeba hosts took in L. pneumophila through a specific and presumably highly efficient uptake mechanism. Living and heat-inactivated P. aeruginosa best supported the replication of L. pneumophila in N. lovaniensis and A. castellanii, respectively, whereas for both amoeba species, E. coli yielded the lowest number of replicated L. pneumophila. Furthermore, microscopic examination showed that 100% of the A. castellanii and only 2% of the N. lovaniensis population were infected with L. pneumophila at the end of the experiment. This study clearly shows the influence of some non-Legionella bacteria on the intracellular replication of L. pneumophila in A. castellanii and N. lovaniensis. It also demonstrates the different abilities of the two tested amoeba species to serve as a proper host for the replication and distribution of the human pathogen in man-made aquatic environments such as cooling towers, shower heads, and air conditioning systems with potential serious consequences for human health.

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References

  1. Y Abu Kwaik (2000) ArticleTitleInvasion of mammalian and protozoan cells by Legionella pneumophila Subcell Biochem 33 383–407

    Google Scholar 

  2. JF Jonckheere ParticleDe (2002) ArticleTitleA century of research on the amoeboflagellate genus Naegleria Acta Protozool 41 309–342

    Google Scholar 

  3. BS Fields (1996) ArticleTitleThe molecular ecology of legionellae Trends Microbiol 4 286–290 Occurrence Handle8829338 Occurrence Handle10.1016/0966-842X(96)10041-X Occurrence Handle1:STN:280:DyaK28vislSruw%3D%3D

    Article  PubMed  CAS  Google Scholar 

  4. BS Fields RF Benson RE Besser (2002) ArticleTitle Legionella and legionnaires' disease: 25 years of investigation Clin Microbiol Rev 15 506–526 Occurrence Handle12097254 Occurrence Handle10.1128/CMR.15.3.506-526.2002

    Article  PubMed  Google Scholar 

  5. G Greub D Raoult (2004) ArticleTitleMicroorganisms resistant to free-living amoebae Clin Microbiol Rev 17 413–433 Occurrence Handle15084508 Occurrence Handle10.1128/CMR.17.2.413-433.2004

    Article  PubMed  Google Scholar 

  6. D Grimm H Merkert W Ludwig KH Schleifer J Hacker BC Brand (1998) ArticleTitleSpecific detection of Legionella pneumophila: construction of a new 16S rRNA-targeted oligonucleotide probe Appl Environ Microbiol 64 2686–2690 Occurrence Handle9647849 Occurrence Handle1:CAS:528:DyaK1cXkt1Ogs7Y%3D

    PubMed  CAS  Google Scholar 

  7. EP Holden HH Winkler DO Wood ED Leinbach (1984) ArticleTitleIntracellular growth of Legionella pneumophila within Acanthamoeba castellanii Neff Infect Immun 45 18–24 Occurrence Handle6376354 Occurrence Handle1:STN:280:DyaL2c3jvVenuw%3D%3D

    PubMed  CAS  Google Scholar 

  8. Jack, DL, Turner, MW (2003) Anti-microbial activities of mannose-binding lectin. Biochem Soc T 31: part 4

    Google Scholar 

  9. K Jürgens J Pernthaler S Schalla R Amann (1999) ArticleTitleMorphological and compositional changes in a planktonic bacterial community in response to enhanced protozoan grazing Appl Environ Microbiol 65 1241–1250 Occurrence Handle10049890

    PubMed  Google Scholar 

  10. CH King BS Fields EB Shotts EH White (1991) ArticleTitleEffects of cytochalasin-D and methylamine on intracellular growth of Legionella pneumophila in amoebas and human monocyte-like cells Infect Immun 59 758–763 Occurrence Handle1997428 Occurrence Handle1:STN:280:DyaK3M7kslaisw%3D%3D

    PubMed  CAS  Google Scholar 

  11. W Manz R Amann R Szewzyk U Szewzyk TA Stenström P Hutzler KH Schleifer (1995) ArticleTitle In situ identification of Legionellaceae using 16S rRNA targeted oligonucleotide probes and confocal laser scanning microscopy Microbiology 141 39 Occurrence Handle10.1099/00221287-141-1-29

    Article  Google Scholar 

  12. L Margolis GW Esch JC Holmes AM Kuris GA Schad (1982) ArticleTitleThe use of ecological terms in parasitology (report of an ad hoc committee of the American Society of Parasitologists) J Parasitol 68 131–133

    Google Scholar 

  13. McNealy, T, Newsome, AL, Johnson, RA, Berk, SG (2002) Impact of amoebae, bacteria and tetrahymena on legionella pneumophila multiplication and distribution in an aquatic environment. In: Proceedings of the Vth International Conference on Legionella, pp 170–175

  14. R Michel H Burghardt H Bergmann (1995) ArticleTitle Acanthamoeba, naturally infected with pseudomonas aeruginosa, after their isolation from a microbiologically contaminated drinking water system in a hospital Zentbl Hyg Umweltmed 196 532–544 Occurrence Handle1:STN:280:DyaK2MzksVCgsw%3D%3D

    CAS  Google Scholar 

  15. JF Moffat (1992) ArticleTitleA quantitative model of intracellular growth of Legionella pneumophila in Acanthamoeba castellanii Infect Immun 60 296–301 Occurrence Handle1729191 Occurrence Handle1:CAS:528:DyaK38XhtVantbs%3D

    PubMed  CAS  Google Scholar 

  16. R Murga TS Forster E Brown JM Pruckler BS Fields RM Donlan (2001) ArticleTitleRole of biofilms in the survival of Legionella pneumophila in a model potable-water system Microbiology 147 3121–3126 Occurrence Handle11700362 Occurrence Handle1:CAS:528:DC%2BD3MXos1ejtb0%3D

    PubMed  CAS  Google Scholar 

  17. JA Noble DG Ahearn SV Avery SA Crow (2002) ArticleTitlePhagocytosis affects biguanide sensitivity of Acanthamoeba spp Antimicrob Agents Chemother 46 2069–2076 Occurrence Handle12069957 Occurrence Handle10.1128/AAC.46.7.2069-2076.2002 Occurrence Handle1:CAS:528:DC%2BD38XltVWntbc%3D

    Article  PubMed  CAS  Google Scholar 

  18. M Pirehma K Suresh S Sivanandam AK Anuar K Ramakrishnan GS Kumar (1999) ArticleTitleField's stain—a rapid staining method for Acanthamoeba spp Parasitol Res 85 791–793 Occurrence Handle10494803 Occurrence Handle10.1007/s004360050634 Occurrence Handle1:STN:280:DyaK1Mvit12qtg%3D%3D

    Article  PubMed  CAS  Google Scholar 

  19. S Rodriguez-Zaragosa (1994) ArticleTitleEcology of free-living amoebae Crit Rev Microbiol 3 225–241

    Google Scholar 

  20. R Rønn AE Mcgraig BS Griffiths JI Prosser (2002) ArticleTitleImpact of protozoan grazing on bacterial community structure in soil microcosms Appl Environ Microbiol 68 6094–6105 Occurrence Handle12450833

    PubMed  Google Scholar 

  21. M Steinert U Hentschel J Hacker (2002) ArticleTitle Legionella pneumophila: an aquatic microbe goes astray FEMS Microbiol Rev 26 149–162 Occurrence Handle12069880 Occurrence Handle10.1016/S0168-6445(02)00093-1 Occurrence Handle1:CAS:528:DC%2BD38XksFymtLk%3D

    Article  PubMed  CAS  Google Scholar 

  22. SB Surman LHG Morton CW Keevil (1994) ArticleTitleThe dependence of Legionella pneumophila on other aquatic bacteria for survival on R2A medium Int Biodeterior Biodegrad 33 223–236 Occurrence Handle10.1016/0964-8305(94)90062-0

    Article  Google Scholar 

  23. S Toze M Cahill LI Sly JA Fuerst (1994) ArticleTitleThe effect of Aeromonas strains on the growth of Legionella J Appl Bacteriol 77 169–174

    Google Scholar 

  24. RL Tyndall EL Domingue (1982) ArticleTitleCocultivation of Legionella pneumophila and free-living amoebae Appl Environ Microbiol 44 954–959 Occurrence Handle7149720 Occurrence Handle1:STN:280:DyaL3s%2FntlCisA%3D%3D

    PubMed  CAS  Google Scholar 

  25. C Venkataraman LY Gao S Bondada Y Abu Kwaik (1998) ArticleTitleIdentification of putative cytoskeletal protein homologues in the protozoan host Hartmannella vermiformis as substrates for induced tyrosine phosphatase activity upon attachment to the Legionnaires' disease bacterium, Legionella pneumophila J Exp Med 188 505–514 Occurrence Handle9687528 Occurrence Handle10.1084/jem.188.3.505 Occurrence Handle1:CAS:528:DyaK1cXltFyjsrY%3D

    Article  PubMed  CAS  Google Scholar 

  26. RM Wadowsky RB Yee (1983) ArticleTitleSatellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve Appl Environ Microbiol 46 1447–1449 Occurrence Handle6660882 Occurrence Handle1:STN:280:DyaL2c7gtlWrtQ%3D%3D

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was funded by the IWT (GBOU no. 20153). The authors wish to thank Joost Raeymaekers, Eugène Rurangwa, Jochen Vandekerkhove, Joost Van Overbeke, and Nico Vromant for their assistance in statistical analyses and the first revision of the manuscript. The authors also acknowledge Suzanne Zweizig for the careful language revision of the text.

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Authors and Affiliations

  1. Laboratory of Aquatic Ecology, Zoological Institute, Katholieke Universiteit Leuven, Charles De Bériotstraat 32, 3000, Louvain, Belgium

    P. Declerck, J. Behets, Y. Delaedt & F. Ollevier

  2. Laboratory of Photochemistry and Spectroscopy, Dept. of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001, Louvain, Belgium

    A. Margineanu

  3. Laboratory of Bacteriology, Rega Institute, Katholieke Universiteit Leuven, Minderbroedersstraat 10, 3000, Louvain, Belgium

    E. Lammertyn

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  1. P. Declerck
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  2. J. Behets
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  3. Y. Delaedt
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  4. A. Margineanu
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Correspondence to P. Declerck.

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Declerck, P., Behets, J., Delaedt, Y. et al. Impact of Non-Legionella Bacteria on the Uptake and Intracellular Replication of Legionella pneumophila in Acanthamoeba castellanii and Naegleria lovaniensis . Microb Ecol 50, 536–549 (2005). https://doi.org/10.1007/s00248-005-0258-0

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  • DOI: https://doi.org/10.1007/s00248-005-0258-0

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