1887

Abstract

Three Legionella -like strains, designed km488, km489 and km521, were isolated from freshwater samples in China. Cells were Gram-stain-negative, rod-shaped and non-spore-forming. Growth was observed on BCYEα agar, but not on BCYEα agar without l-cysteine, chocolate agar with PolyViteX or Columbia blood agar. The major fatty acids (>5 %) of strains km488, km489 and km521 were C16 : 0, anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The mip gene sequences (574 nt) showed the isolates were almost identical with more than 99.7 % sequence similarities, and closely matched to L. gormanii ATCC 33297 with 95.4–95.6 % sequence similarities. Phylogenetic analyses based on concatenated gene (16S rRNA, mip, rpoB and rnpB) sequences indicated that the isolates formed a distinct cluster along with L. gormanii within the genus Legionella . Matrix-assisted laser desorption ionization time-of-flight analyses also demonstrated a clear separation between the isolates and other closely and distantly related Legionella species. DNA–DNA hybridization studies demonstrated that the isolates were closely related (92.0 –95.0 % DNA-DNA relatedness) but differentiated from their phylogenetic neighbours (<70 % DNA–DNA relatedness). The whole genome of km488 was sequenced, and showed a G+C content of 37.8 mol%. Based on the findings from this polyphasic taxonomic study, the isolates are considered to represent a single novel species, for which the name Legionella qingyii sp. nov. is proposed. The type strain is km488 (KCTC 15636=CCTCC AB 2018025=NRBC 113223).

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2019-05-07
2021-08-04
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References

  1. Swart AL, Harrison CF, Eichinger L, Steinert M, Hilbi H. Acanthamoeba and Dictyostelium as Cellular Models for Legionella Infection. Front Cell Infect Microbiol 2018; 8:61 [View Article][PubMed]
    [Google Scholar]
  2. Burillo A, Pedro-Botet ML, Bouza E. Microbiology and epidemiology of legionnaire's disease. Infect Dis Clin North Am 2017; 31:7–27 [View Article][PubMed]
    [Google Scholar]
  3. Fields BS, Benson RF, Besser RE. Legionella and Legionnaires disease: 25 years of investigation. Clin Microbiol Rev 2002; 15:506–526 [View Article][PubMed]
    [Google Scholar]
  4. Newton HJ, Ang DK, van Driel IR, Hartland EL. Molecular pathogenesis of infections caused by Legionella pneumophila . Clin Microbiol Rev 2010; 23:274–298 [View Article][PubMed]
    [Google Scholar]
  5. Gomez-Valero L, Buchrieser C. Genome dynamics in Legionella: the basis of versatility and adaptation to intracellular replication. Cold Spring Harb Perspect Med 2013; 3:297–316 [View Article][PubMed]
    [Google Scholar]
  6. LPSN List of prokaryotic names with standing in nomenclature Legionella . Available at http://www.bacterio.net/legionella.html
  7. Cunha BA, Burillo A, Bouza E. Legionnaires' disease. The Lancet 2016; 387:376–385 [View Article]
    [Google Scholar]
  8. Palmer A, Painter J, Hassler H, Richards VP, Bruce T et al. Legionella clemsonensis sp. nov.: a green fluorescing Legionella strain from a patient with pneumonia. Microbiol Immunol 2016; 60:694–701 [View Article][PubMed]
    [Google Scholar]
  9. Relich RF, Schmitt BH, Raposo H, Barker L, Blosser SJ et al. Legionella indianapolisensis sp. nov., isolated from a patient with pulmonary abscess. Int J Infect Dis 2018; 69:26–28 [View Article][PubMed]
    [Google Scholar]
  10. Xiao-Yong Z, Chao-Hui H, Qing-Yi Z. Comparative study on sampling methods for monitoring Legionella species in environmental water. Afr J Microbiol Res 2014; 8:974–985 [View Article]
    [Google Scholar]
  11. James G. Universal bacterial identification by PCR and DNA sequencing of 16S rRNA gene [M]. In Carter IWJ, Schuller M, James GS. (editors) PCR for Clinical Microbiology Verlag Ny: Springer; 2010
    [Google Scholar]
  12. Ratcliff RM, Lanser JA, Manning PA, Heuzenroeder MW. Sequence-based classification scheme for the genus Legionella targeting the mip gene. J Clin Microbiol 1998; 36:1560–1567[PubMed]
    [Google Scholar]
  13. Ko KS, Lee HK, Park MY, Lee KH, Yun YJ et al. Application of RNA polymerase beta-subunit gene (rpoB) sequences for the molecular differentiation of Legionella species. J Clin Microbiol 2002; 40:2653–2658[PubMed]
    [Google Scholar]
  14. Rubin CJ, Thollesson M, Kirsebom LA, Herrmann B. Phylogenetic relationships and species differentiation of 39 Legionella species by sequence determination of the RNase P RNA gene rnpB . Int J Syst Evol Microbiol 2005; 55:2039–2049 [View Article][PubMed]
    [Google Scholar]
  15. Yang Z. Phylogenetic analysis using parsimony and likelihood methods. J Mol Evol 1996; 42:294–307 [View Article][PubMed]
    [Google Scholar]
  16. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987; 4:406 [View Article][PubMed]
    [Google Scholar]
  17. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 2016; 33:1870–1880 [View Article][PubMed]
    [Google Scholar]
  18. Edelstein PH, Edelstein MA. Comparison of the plating efficiencies and shelf lives of three different commercial buffered charcoal yeast extract media supplemented with alpha-ketoglutaric acid. J Clin Microbiol 2010; 48:1882–1883 [View Article][PubMed]
    [Google Scholar]
  19. Brenner DJ, Steigerwalt AG, Gorman GW, Wilkinson HW, Bibb WF et al. Ten new species of Legionella . Int J Syst Bacteriol 1985; 35:50–59 [View Article]
    [Google Scholar]
  20. Humble MW, King A, Phillips I. API ZYM: a simple rapid system for the detection of bacterial enzymes. J Clin Pathol 1977; 30:275–277 [View Article][PubMed]
    [Google Scholar]
  21. Lambert MA, Moss CW. Cellular fatty acid compositions and isoprenoid quinone contents of 23 Legionella species. J Clin Microbiol 1989; 27:465–473[PubMed]
    [Google Scholar]
  22. He Y, Chang TC, Li H, Shi G, Tang YW. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and database for identification of Legionella species. Can J Microbiol 2011; 57:533–538 [View Article][PubMed]
    [Google Scholar]
  23. Ezaki T, Hashimoto Y, Yabuuchi E. Fluorometric deoxyribonucleic acid-deoxyribonucleic acid hybridization in microdilution wells as an alternative to membrane filter hybridization in which radioisotopes are used to determine genetic relatedness among bacterial strains. Int J Syst Bacteriol 1989; 39:224–229 [View Article]
    [Google Scholar]
  24. Ezaki T, Hashimoto Y, Yamamoto H, Lucida ML, Liu SL et al. Evaluation of the microplate hybridization method for rapid identification of Legionella species. Eur J Clin Microbiol Infect Dis 1990; 9:213–217 [View Article][PubMed]
    [Google Scholar]
  25. Stackebrandt E, Goebel BM. Taxonomic Note: A place for dna-dna reassociation and 16s rrna sequence analysis in the present species definition in bacteriology. Int J Syst Evol Microbiol 1994; 44:846–849 [View Article]
    [Google Scholar]
  26. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014; 30:2114–2120 [View Article][PubMed]
    [Google Scholar]
  27. Zerbino DR, Birney E. Velvet: algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 2008; 18:821–829 [View Article][PubMed]
    [Google Scholar]
  28. Boetzer M, Henkel CV, Jansen HJ, Butler D, Pirovano W. Scaffolding pre-assembled contigs using SSPACE. Bioinformatics 2011; 27:578–579 [View Article][PubMed]
    [Google Scholar]
  29. Li W, Godzik A. Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences. Bioinformatics 2006; 22:1658–1659 [View Article][PubMed]
    [Google Scholar]
  30. Dennis PJ, Brenner DJ, Thacker WL, Wait R, Vesey G et al. Five new Legionella species isolated from water. Int J Syst Bacteriol 1993; 43:329–337 [View Article][PubMed]
    [Google Scholar]
  31. Chun J, Oren A, Ventosa A, Christensen H, Arahal DR et al. Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 2018; 68:461–466 [View Article][PubMed]
    [Google Scholar]
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