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Role of Biofilm in Protection of the Replicative Form of Legionella pneumophila

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Abstract

The dual nature of Legionella pneumophila enables its survival in free and intracellular environments and underpins its infection and spread mechanisms. Experiments using bacterial cultures and improved RTqPCR protocols were devised to gain fresh insights into the role of biofilm in protecting the replicative form of L. pneumophila. mip gene expression was used as a marker of virulence in sessile (biofilm-bound) and planktonic (free-floating) cells of L. pneumophila serotype 1 ATCC 33152. The ratio of mip gene expression to transcriptionally active Legionella cells increased both in sessile and free-floating cells demonstrating an up-regulation of mip gene under nutrient depletion. However, a different trend was observed between the two forms, in planktonic cells the mip gene expression/transcriptionally active Legionella cells increased until the end of the experiment, while in the biofilm such increase was observed at the end of the experiment. These findings suggest a possible association between the switch to the transmissive phase of Legionella and a mip up-regulation and a role for biofilm in preserving Legionella cells in replicative form. Moreover, it has been shown that improved RTqPCR protocols are valuable tools to explore bacterial virulence.

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References

  1. Brüggemann H, Hagman A, Jules M, Sismeiro O, Dillies MA, Gouyette C, Kunst F, Steinert M, Heuner K, Coppée JY, Buchrieser C (2006) Virulence strategies for infecting phagocytes deduced from the in vivo transcriptional program of Legionella pneumophila. Cell Microbiol 8:1228–1240

    Article  PubMed  Google Scholar 

  2. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622

    Article  PubMed  CAS  Google Scholar 

  3. Cianciotto NP, Eisenstein BI, Mody CH, Engleberg NC (1990) A mutation in the mip gene results in an attenuation of Legionella pneumophila virulence. J Infect Dis 162:121–126

    Article  PubMed  CAS  Google Scholar 

  4. Cianciotto NP, Fields BS (1992) Legionella pneumophila mip gene potentiates intracellular infection of protozoa and human macrophages. Proc Natl Acad Sci U S A 89:5188–5191

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  5. Declerck P, Behets J, Margineanu A, van Hoef V, De Keersmaecker B, Ollevier F (2009) Replication of Legionella pneumophila in biofilms of water distribution pipes. Microbiol Res 164:593–603

    Article  PubMed  CAS  Google Scholar 

  6. Di Cesare A, Luna GM, Vignaroli C, Pasquaroli S, Tota S, Paroncini P, Biavasco F (2013) Aquaculture can promote the presence and spread of antibiotic-resistant Enterococci in marine sediments. PLoS ONE 8:e62838

    Article  PubMed  PubMed Central  Google Scholar 

  7. Donlan RM, Costerton JW (2002) Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15:167–193

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  8. Fields BS, Benson RF, Besser RE (2002) Legionella and Legionnaires’ disease: 25 years of investigation. Clin Microbiol Rev 15:506–526

    Article  PubMed  PubMed Central  Google Scholar 

  9. Hindré T, Brüggemann H, Buchrieser C, Héchard Y (2008) Transcriptional profiling of Legionella pneumophila biofilm cells and the influence of iron on biofilm formation. Microbiology 154:30–41

    Article  PubMed  Google Scholar 

  10. Huws SA, McBain AJ, Gilbert P (2005) Protozoan grazing and its impact upon population dynamics in biofilm communities. J Appl Microbiol 98:238–244

    Article  PubMed  CAS  Google Scholar 

  11. Kuiper MW, Wullings BA, Akkermans AD, Beumer RR, van der Kooij D (2004) Intracellular proliferation of Legionella pneumophila in Hartmannella vermiformis in aquatic biofilms grown on plasticized polyvinyl chloride. Appl Environ Microbiol 70:6826–6833

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  12. Molofsky AB, Swanson MS (2003) Legionella pneumophila CsrA is a pivotal repressor of transmission traits and activator of replication. Mol Microbiol 50:445–461

    Article  PubMed  CAS  Google Scholar 

  13. Molofsky AB, Swanson MS (2004) Differentiate to thrive: lessons from the Legionella pneumophila life cycle. Mol Microbiol 53:29–40

    Article  PubMed  CAS  Google Scholar 

  14. Murga R, Forster TS, Brown E, Pruckler JM, Fields BS, Donlan RM (2001) Role of biofilms in the survival of Legionella pneumophila in a model potable-water system. Microbiology 147:3121–3126

    PubMed  CAS  Google Scholar 

  15. Nomanpour B, Ghodousi A, Babaei T, Jafari S, Feizabadi MM (2012) Single tube real time PCR for detection of Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila from clinical samples of CAP. Acta Microbiol Immunol Hung 59:171–184

    Article  PubMed  CAS  Google Scholar 

  16. Piao Z, Sze CC, Barysheva O, Iida K, Yoshida S (2006) Temperature-regulated formation of mycelial mat-like biofilms by Legionella pneumophila. Appl Environ Microbiol 72:1613–1622

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  17. Price CT, Richards AM, Von Dwingelo JE, Samara HA, Abu Kwaik Y (2014) Amoeba host-Legionella synchronization of amino acid auxotrophy and its role in bacterial adaptation and pathogenic evolution. Environ Microbiol 16:350–358

    Article  PubMed  CAS  Google Scholar 

  18. Qasem JA, Mustafa AS, Khan ZU (2008) Legionella in clinical specimens and hospital water supply facilities: molecular detection and genoty** of the isolates. Med Princ Pract 17:49–55

    Article  PubMed  CAS  Google Scholar 

  19. Rogers J, Dowsett AB, Dennis PJ, Lee JV, Keevil CW (1994) Influence of temperature and plumbing material selection on biofilm formation and growth of Legionella pneumophila in a model potable water system containing complex microbial flora. Appl Environ Microbiol 60:1585–1592

    PubMed  CAS  PubMed Central  Google Scholar 

  20. Steinert M, Hentschel U, Hacker J (2002) Legionella pneumophila: an aquatic microbe goes astray. FEMS Microbiol Rev 26:149–162

    Article  PubMed  CAS  Google Scholar 

  21. Temmerman R, Vervaeren H, Noseda B, Boon N, Verstraete W (2006) Necrotrophic growth of Legionella pneumophila. Appl Environ Microbiol 72:4323–4328

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  22. Wagner C, Khan AS, Kamphausen T, Schmausser B, Unal C, Lorenz U, Fischer G, Hacker J, Steinert M (2007) Collagen binding protein Mip enables Legionella pneumophila to transmigrate through a barrier of NCI-H292 lung epithelial cells and extracellular matrix. Cell Microbiol 9:450–462

    Article  PubMed  CAS  Google Scholar 

  23. Walser SM, Gerstner DG, Brenner B, Höller C, Liebl B, Herr CE (2014) Assessing the environmental health relevance of cooling towers—a systematic review of legionellosis outbreaks. Int J Hyg Environ Healt 217(2–3):145–154

    Article  Google Scholar 

  24. Watnick P, Kolter R (2000) Biofilm, city of microbes. J Bacteriol 182:2675–2679

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  25. Wieland H, Faigle M, Lang F, Northoff H, Neumeister B (2002) Regulation of the Legionella mip-promotor during infection of human monocytes. FEMS Microbiol Lett 212:127–132

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biomolecular Sciences, Section of Toxicological, Hygienistic and Environmental Sciences, University of Urbino “Carlo Bo”, Urbino, Italy

    Elisa Andreozzi, Luigia Sabatini, Elisa Chessa & Barbara Citterio

  2. Department of Life and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy

    Andrea Di Cesare

  3. Microbial Ecology Group, CNR – Institute of Ecosystem Study, Verbania, Italy

    Andrea Di Cesare

  4. Department of Biomolecular Sciences, Section of Pharmacology and Pharmacognosy, University of Urbino “Carlo Bo”, Urbino, Italy

    Davide Sisti & Marco Rocchi

Authors
  1. Elisa Andreozzi
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  2. Andrea Di Cesare
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  3. Luigia Sabatini
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  4. Elisa Chessa
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  5. Davide Sisti
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  6. Marco Rocchi
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  7. Barbara Citterio
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Correspondence to Barbara Citterio.

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Andreozzi, E., Di Cesare, A., Sabatini, L. et al. Role of Biofilm in Protection of the Replicative Form of Legionella pneumophila . Curr Microbiol 69, 769–774 (2014). https://doi.org/10.1007/s00284-014-0648-y

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  • DOI: https://doi.org/10.1007/s00284-014-0648-y

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