Molecular diversity and high virulence of Legionella pneumophila strains isolated from biofilms developed within a warm spring of a thermal spa

BMC Microbiology

Christophe Gilbert¹  Dominique Fontvieille³  Danièle Atlan¹  Sophie Jarraud²  Monique Reyrolle⁴  Françoise Forey⁴  Zineddine Chaabna³ 

[1] Université de Lyon, CNRS UMR 5240, Université Lyon1, Villeurbanne, F-69622, France;Université Lyon, INSERM U851, Université Lyon 1, Lyon, F-69008, France;UMR CARRTEL, Université de Savoie-INRA, Le Bourget du Lac, F-73376, France;Centre de Biologie et Pathologie Est, Centre National de Référence des Légionelles, Bron, F-69677, France

关键词: Virulence;    Biodiversity;    Warm spring;    Biofilms;    Legionella pneumophila;   

Others  :  1144547
DOI  :  10.1186/1471-2180-13-17

 received in 2012-07-24, accepted in 2013-01-14,  发布年份 2013

【 摘 要 】

Background

Several cases of legionellosis have been diagnosed in the same French thermal spa in 1986, 1994 and 1997. L. pneumophila serogroup 1 (Lp1) strains have been isolated from several patients, but the source of contamination was not identified despite the presence of different Lp1 in water samples of the three natural springs feeding the spa at this period.

Results

Our strategy was to investigate L. pneumophila (Lp) strains from natural biofilms developed in a sulphur-rich warm spring of this contaminated site. Biofilm analysis revealed the presence of three Lp serogroups (Lp1, Lp10 and Lp12). Surprisingly, Lp10 and Lp12 were not reported in the previous described studies from water samples. Besides, the new seven Lp1 we isolated exhibit a high molecular diversity and have been differentiated in five classes according to their DNA genome patterns obtained by PFGE and mip sequences. It must be noted that these DNA patterns are original and unknown in databases. Interestingly, the 27 Lp environmental strains we isolated display a higher cytotoxicity and virulence towards the amoeba Acanthamoeba castellanii than those of known Lp1 epidemic strains.

Conclusion

The characteristics of Legionella pneumophila Lp1 strains isolated from the warm spring are in agreement with their presence in biofilms and their probable long-term persistence in this ecosystem.

【 授权许可】

   
2013 Chaabna et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】

• [1]McDade JE, Shepard CC, Fraser DW, Tsai TR, Redus MA, Dowdle WR: Legionnaires’ Disease. N Engl J Med 1977, 297(22):1197-1203.
• [2]Nguyen TMN, Ilef D, Jarraud S, Rouil L, Campese C, Che D, Haeghebaert S, Ganiayre FO, Marcel F, Etienne J, et al.: A community-wide outbreak of legionnaires disease linked to industrial cooling towers - How far can contaminated aerosols spread? J Infect Dis 2006, 193(1):102-111.
• [3]Rowbotham TJ: Preliminary report on the pathogenicity of Legionella pneumophila for freshwater and soil amoebae. J Clin Pathol 1980, 33(12):1179-1183.
• [4]Doleans A, Aurell H, Reyrolle M, Lina G, Freney J, Vandenesch F, Etienne J, Jarraud S: Clinical and environmental distributions of Legionella strains in France are different. J Clin Microbiol 2004, 42(1):458-460.
• [5]Gomez-Valero L, Rusniok C, Buchrieser C: Legionella pneumophila: population genetics, phylogeny and genomics. Infect Genet Evol 2009, 9(5):727-739.
• [6]Yu VL, Plouffe JF, Pastoris MC, Stout JE, Schousboe M, Widmer A, Summersgill J, File T, Heath CM, Paterson DL, et al.: Distribution of Legionella species and serogroups isolated by culture in patients with sporadic community-acquired legionellosis: An international collaborative survey. J Infect Dis 2002, 186(1):127-128.
• [7]Cazalet C, Rusniok C, Bruggemann H, Zidane N, Magnier A, Ma L, Tichit M, Jarraud S, Bouchier C, Vandenesch F, et al.: Evidence in the Legionella pneumophila genome for exploitation of host cell functions and high genome plasticity. Nat Genet 2004, 36(11):1165-1173.
• [8]Albert-Weissenberger C, Cazalet C, Buchrieser C: Legionella pneumophila - a human pathogen that co-evolved with fresh water protozoa. Cell Mol Life Sci 2007, 64:432-448.
• [9]Isberg RR, O’Connor TJ, Heidtman M: The Legionella pneumophila replication vacuole: making a cosy niche inside host cells. Nat Rev Microbiol 2009, 7(1):12-24.
• [10]Hilbi H, Jarraud S, Hartland E, Buchrieser C: Update on Legionnaires’ disease: pathogenesis, epidemiology, detection and control. Mol Microbiol 2010, 76(1):1-11.
• [11]Merault N, Rusniok C, Jarraud S, Gomez-Valero L, Cazalet C, Marin M, Brachet E, Aegerter P, Gaillard JL, Etienne J, et al.: Specific real-time PCR for simultaneous detection and identification of Legionella pneumophila serogroup 1 in water and clinical samples. Appl Environ Microbiol 2011, 77(5):1708-1717.
• [12]Bornstein N, Marmet D, Surgot M, Nowicki M, Arslan A, Esteve J, Fleurette J: Exposure to Legionellaceae at a hot-spring spa - a prospective clinical and serological study. Epidemiol Infect 1989, 102(1):31-36.
• [13]Molmeret M, Jarraud S, Morin JP, Pernin P, Forey F, Reyrolle M, Vandenesch F, Etienne J, Farge P: Different growth rates in amoeba of genotypically related environmental and clinical Legionella pneumophila strains isolated from a thermal spa. Epidemiol Infect 2001, 126(2):231-239.
• [14]Bandyopadhyay P, Steinman HM: Legionella pneumophila catalase-peroxidases: cloning of the katB gene and studies of KatB function. J Bacteriol 1998, 180(20):5369-5374.
• [15]Bandyopadhyay P, Steinman HM: Catalase-peroxidases of Legionella pneumophila: cloning of the katA gene and studies of KatA function. J Bacteriol 2000, 182(23):6679-6686.
• [16]Pine L, Hoffman PS, Malcolm GB, Benson RF, Gorman GW: Whole-cell peroxidase test for identification of Legionella pneumophila. J Clin Microbiol 1984, 19(2):286-290.
• [17]Ratcliff RM, Lanser JA, Manning PA, Heuzenroeder MW: Sequence-based classification scheme for the genus Legionella targeting the mip gene. J Clin Microbiol 1998, 36(6):1560-1567.
• [18]Yong SFY, Tan SH, Wee J, Tee JJ, Sansom FM, Newton HJ, Hartland EL: Molecular detection of Legionella: moving on from mip. Front Microbiol 2010, 1:123.
• [19]Ferhat M, Atlan D, Vianney A, Lazzaroni JC, Doublet P, Gilbert C: The TolC protein of Legionella pneumophila plays a major role in multi-drug resistance and the early steps of host invasion. PLoS One 2009., 4(11)
• [20]Cirillo JD, Cirillo SLG, Yan L, Bermudez LE, Falkow S, Tompkins LS: Intracellular growth in Acanthamoeba castellanii affects monocyte entry mechanisms and enhances virulence of Legionella pneumophila. Infect Immun 1999, 67(9):4427-4434.
• [21]Neumeister B, Reiff G, Faigle M, Dietz K, Northoff H, Lang F: Influence of Acanthamoeba castellanii on intracellular growth of different Legionella species in human monocytes. Appl Environ Microbiol 2000, 66(3):914-919.
• [22]Thacker WL, Wilkinson HW, Benson RF, Brenner DJ: Legionella-pneumophila serogroup-12 isolated from human and environmental sources. J Clin Microbiol 1987, 25(3):569-570.
• [23]Amaro F, Gilbert JA, Owens S, Trimble W, Shuman HA: Whole-genome sequence of the human pathogen Legionella pneumophila serogroup 12 strain 570-CO-H. J Bacteriol 2012, 194(6):1613-1614.
• [24]Lamarque M, Aubel D, Piard JC, Gilbert C, Juillard V, Atlan D: The peptide transport system Opt is involved in both nutrition and environmental sensing during growth of Lactococcus lactis in milk. Microbiology-Sgm 2011, 157:1612-1619.
• [25]Reyrolle M, Ratat C, Leportier M, Jarraud S, Freney J, Etienne J: Rapid identification of Legionella pneumophila serogroups by latex agglutination. Eur J Clin Microbiol Infect Dis 2004, 23(11):864-866.
• [26]Lawrence C, Reyrolle M, Dubrou S, Forey F, Decludt B, Goulvestre C, Matsiota-Bernard P, Etienne J, Nauciel C: Single clonal origin of a high proportion of Legionella pneumophila serogroup 1 isolates from patients and the environment in the area of Paris, France, over a 10-year period. J Clin Microbiol 1999, 37(8):2652-2655.