1887

Abstract

-like isolates, strains 27fs60, 30fs61 and 30cs62, were isolated from a hotel water distribution system in the Emilia-Romagna region, Italy. Isolates were Gram- and Ziehl Neelsen-stain-negative, rod-shaped, with transitory flagella presence and able to grow at 32–37 °C (with an optimum at 32 °C) on buffered charcoal–yeast extract agar with -cysteine, glycine–vancomycin–polymyxin B–cycloheximide agar and Wadowsky–Yee medium agar. The strains showed positive reactions for oxidase, hippurate and gelatinase and a weakly positive reaction for catalase. Based on the EUCAST cut-off, strain 30cs62 was resistant to ciprofloxacin (5 mg l). The and gene sequences of the three strains showed close matches to those of ATCC 49507 with similarity values of 98.2 and 94.5 %, respectively. Whole genome sequencing of the three strains was performed, resulting in G+C contents of 39.0, 39.1 and 39.0 mol%, respectively. The identity percentage measured by average nucleotide identity between the three strains and their respective closest strains were: 91.32 % . NCTC 12376, 91.45 % . ATCC 49507 and 91.45 % . ATCC 49507, respectively. The digital DNA–DNA hybridization analysis demonstrated how the isolates were separated from the most related phylogenetic species ( ATCC 49507, ≤40.10 % DNA–DNA relatedness). The concatenated phylogenetic tree based on 16S , and genes, shows a close relationship with ATCC 49507. The results obtained confirm the status of an independent species. The name proposed for this species is sp. nov. with 30cs62 (=ATCC TSD-262=DSM 112526) as the type strain.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/ijsem/10.1099/ijsem.0.005512
2022-09-29
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/ijsem/72/9/ijsem005512.html?itemId=/content/journal/ijsem/10.1099/ijsem.0.005512&mimeType=html&fmt=ahah

References

  1. Newton HJ, Ang DKY, van Driel IR, Hartland EL. Molecular pathogenesis of infections caused by Legionella pneumophila. Clin Microbiol Rev 2010; 23:274–298 [View Article]
    [Google Scholar]
  2. Parte AC, Road W. LPSN–List of Prokaryotic Names with Standing in Nomenclature. Nucleic Acids Res 2014; 42:D613–6 [View Article]
    [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. Rota MC, Caporali MG, Bella A, Scaturro M, Giannitelli S et al. The legionellosis surveillance system in Italy: 2019 results. Boll Epidemiol Naz 2020; 1:32–38
    [Google Scholar]
  5. Centers for Disease Control and Prevention Legionella (Legionnaires’ Disease and Pontiac Fever) – Causes, How it Spreads, and People at Increased Risk; 2020 www.cdc.gov/legionella/about/causes-transmission.html
  6. Correia AM, Ferreira JS, Borges V, Nunes A, Gomes B et al. 'Probable person-to-person transmission of Legionnaires’ disease. N Engl J Med 2016; 374:497–498 [View Article] [PubMed]
    [Google Scholar]
  7. Borges V, Nunes A, Sampaio DA, Vieira L, Machado J et al. Legionella pneumophila strain associated with the first evidence of person-to-person transmission of Legionnaires’ disease: a unique mosaic genetic backbone. Sci Rep 2016; 6:1–11 [View Article]
    [Google Scholar]
  8. ISO UNI EN ISO 19458:2006 – Water quality — Sampling for microbiological analysis. Geneva, Switzerland: ISO; 2006 www.iso.org/standard/33845.html
  9. ISO ISO 11731:2017 Water quality – Enumeration of Legionella. Geneva, Switzerland: ISO; 2017 www.iso.org/standard/61782.html
  10. 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 [View Article] [PubMed]
    [Google Scholar]
  11. Ko KS, Lee HK, Park M-Y, Lee K-H, Yun Y-J 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 [View Article] [PubMed]
    [Google Scholar]
  12. Girolamini L, Salaris S, Orsini M, Pascale MR, Mazzotta M et al. Draft genome sequences of Legionella presumptive novel species isolated during environmental surveillance in artificial water systems. Microbiol Resour Announc 2021; 10:e00307-21 [View Article]
    [Google Scholar]
  13. Quijada NM, Rodríguez-Lázaro D, Eiros JM, Hernández M. TORMES: an automated pipeline for whole bacterial genome analysis. Bioinformatics 2019; 35:4207–4212 [View Article]
    [Google Scholar]
  14. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012; 19:455–477 [View Article] [PubMed]
    [Google Scholar]
  15. Chen K-T, Liu C-L, Huang S-H, Shen H-T, Shieh Y-K et al. CSAR: a contig scaffolding tool using algebraic rearrangements. Bioinformatics 2018; 34:109–111 [View Article] [PubMed]
    [Google Scholar]
  16. Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M et al. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 2012; 28:1647–1649 [View Article] [PubMed]
    [Google Scholar]
  17. Tatusova T, DiCuccio M, Badretdin A, Chetvernin V, Nawrocki EP et al. NCBI Prokaryotic Genome Annotation Pipeline. Nucleic Acids Res 2016; 44:6614–6624 [View Article] [PubMed]
    [Google Scholar]
  18. Yoon S-H, Ha S-M, Lim J, Kwon S, Chun J. A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie van Leeuwenhoek 2017; 110:1281–1286 [View Article]
    [Google Scholar]
  19. Jain C, Rodriguez-R LM, Phillippy AM, Konstantinidis KT, Aluru S. High throughput ANI analysis of 90K prokaryotic genomes reveals clear species boundaries. Nat Commun 2018; 9:1–8 [View Article] [PubMed]
    [Google Scholar]
  20. Tanizawa Y, Fujisawa T, Nakamura Y. DFAST: a flexible prokaryotic genome annotation pipeline for faster genome publication. Bioinformatics 2018; 34:1037–1039 [View Article] [PubMed]
    [Google Scholar]
  21. Richter M, Rosselló-Móra R, Oliver Glöckner F, Peplies J. JSpeciesWS: a web server for prokaryotic species circumscription based on pairwise genome comparison. Bioinformatics 2016; 32:929–931 [View Article] [PubMed]
    [Google Scholar]
  22. Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J et al. BLAST+: architecture and applications. BMC Bioinformatics 2009; 10:1–9 [View Article] [PubMed]
    [Google Scholar]
  23. Meier-Kolthoff JP, Auch AF, Klenk HP, Göker M. Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 2013; 14:60 [View Article]
    [Google Scholar]
  24. Kim M, Oh HS, Park SC, Chun J. Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Int J Syst Evol Microbiol 2014; 64:346–351 [View Article] [PubMed]
    [Google Scholar]
  25. Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 2004; 32:1792–1797 [View Article] [PubMed]
    [Google Scholar]
  26. Price MN, Dehal PS, Arkin AP. FastTree 2–approximately maximum-likelihood trees for large alignments. PLoS One 2010; 5:e9490 [View Article]
    [Google Scholar]
  27. EUCAST Antimicrobial Susceptibility Testing of Legionella pneumophila. Vaxjo, Sweden: 2021 www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/General_documents/Legionella_guidance_document_20160909.pdf
  28. Appelt S, Heuner K. The flagellar regulon of Legionella—a review. Front Cell Infect Microbiol 2017; 7:1–13 [View Article]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/ijsem/10.1099/ijsem.0.005512
Loading
/content/journal/ijsem/10.1099/ijsem.0.005512
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error