1887

Abstract

Ten atypical strains were isolated from Italian raw milk cheeses. The 16S rRNA gene, phenylalanyl-tRNA synthase alpha subunit (), RNA polymerase alpha subunit () and the 16S–23S rRNA intergenic transcribed spacer (ITS) sequences, randomly amplified polymorphic DNA (RAPD) PCR and the phenotypic properties revealed that the isolates represent a novel enterococcal species. On the basis of 16S rRNA gene sequence analysis, the isolates were closely related to ATCC 8043, CECT 411 and ATCC 19434, with 98.8, 98.9 and 99.4 % sequence similarity, respectively. On the basis of sequence analysis of the housekeeping gene , the reference strain, BT159, occupied a position separate from LMG 16198. The group of isolates could be easily differentiated from recognized species of the genus by 16S–23S rRNA ITS analysis, RAPD-PCR and phenotypic characteristics. The name sp. nov. is proposed, with BT159 ( = DSM 23655 = LMG 25958) as the type strain.

Funding
This study was supported by the:
  • VALTEC
  • Accordo Quadro CNR-Regione Lombardia
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2012-08-01
2024-04-20
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References

  1. Andrighetto C., Borney F., Barmaz A., Stefanon B., Lombardi A. 2002; Genetic diversity of Streptococcus thermophilus strains isolated from Italian traditional cheese. Int Dairy J 12:141–144 [View Article]
    [Google Scholar]
  2. Botina S. G., Sukhodolets V. V. 2006; Speciation in bacteria: comparison of the 16S rRNA gene for closely related Enterococcus species. Russ J Genet 42:247–251 [View Article]
    [Google Scholar]
  3. CLSI 2007 Performance Standards for Antimicrobial Susceptibility Testing, Seventeenth Informational Supplement. M100–S17 Wayne, PA: NCCLS;
    [Google Scholar]
  4. Collins M. D., Jones D., Farrow J. A. E., Kilpper-Balz R., Schleifer K. H. 1984; Enterococcus avium nom. rev., comb. nov.; E. casseliflavus nom. rev., comb. nov.; E. durans nom. rev., comb. nov.; E. gallinarum comb. nov.; and E. malodoratus sp. nov.. Int J Syst Bacteriol 34:220–223 [View Article]
    [Google Scholar]
  5. Collins M. D., Farrow J. A., Jones D. 1986; Enterococcus mundtii sp. nov.. Int J Syst Bacteriol 36:8–12 [View Article]
    [Google Scholar]
  6. Commission of the European Communities 1993 Certification of the fatty acid profile and mass fraction of butyric acid (n-butanoic acid), cholesterol and three individual sterols of an anhydrous milk fat reference material with values for information on triglycerides, minor fatty acids, added vanillin and ‘total’ sterol mass fraction. CRM 164. Report 15277.
  7. Cremonesi P., Castiglioni B., Malferrari G., Biunno I., Vimercati C., Moroni P., Morandi S., Luzzana M. 2006; Technical note: improved method for rapid DNA extraction of mastitis pathogens directly from milk. J Dairy Sci 89:163–169 [View Article][PubMed]
    [Google Scholar]
  8. Devriese L. A., Pot B. 1995; The genus Enterococcus . In The Genera of Lactic Acid Bacteria pp. 327–367 Edited by Wood B. J. B., Holzapfel W. H. London: Blackie Academic & Professional; [View Article]
    [Google Scholar]
  9. Devriese L. A., Pot B., Collins M. D. 1993; Phenotypic identification of the genus Enterococcus and differentiation of phylogenetically distinct enterococcal species and species groups. J Appl Bacteriol 75:399–408 [View Article][PubMed]
    [Google Scholar]
  10. Edwards U., Rogall T., Blöcker H., Emde M., Böttger E. C. 1989; Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 17:7843–7853 [View Article][PubMed]
    [Google Scholar]
  11. Farrow J. A. E., Collins M. D. 1985; Enterococcus hirae, a new species that includes amino acid assay strain NCDO 1258 and strains causing growth depression in young chickens. Int J Syst Bacteriol 35:73–75 [View Article]
    [Google Scholar]
  12. Fisher K., Phillips C. 2009; The ecology, epidemiology and virulence of Enterococcus . Microbiology 155:1749–1757 [View Article][PubMed]
    [Google Scholar]
  13. Fortina M. G., Ricci G., Mora D., Manachini P. L. 2004; Molecular analysis of artisanal Italian cheeses reveals Enterococcus italicus sp. nov.. Int J Syst Evol Microbiol 54:1717–1721 [View Article][PubMed]
    [Google Scholar]
  14. Foulquié Moreno M. R., Sarantinopoulos P., Tsakalidou E., De Vuyst L. 2006; The role and application of enterococci in food and health. Int J Food Microbiol 106:1–24 [View Article][PubMed]
    [Google Scholar]
  15. Franz C. M., van Belkum M. J., Holzapfel W. H., Abriouel H., Gálvez A. 2007; Diversity of enterococcal bacteriocins and their grouping in a new classification scheme. FEMS Microbiol Rev 31:293–310 [View Article][PubMed]
    [Google Scholar]
  16. Giraffa G. 2003; Functionality of enterococci in dairy products. Int J Food Microbiol 88:215–222 [View Article][PubMed]
    [Google Scholar]
  17. Harrigan W. F. 1998; Reactions involving protein, amino acids and other nitrogen compounds, including tests for proteolytic activity. In Laboratory Methods in Food Microbiology, 3rd edn. pp. 100–107 Edited by Harrigan W. F. London: Academic Press;
    [Google Scholar]
  18. Jensen M. A., Webster J. A., Straus N. 1993; Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol 59:945–952[PubMed]
    [Google Scholar]
  19. Klein G. 2003; Taxonomy, ecology and antibiotic resistance of enterococci from food and the gastro-intestinal tract. Int J Food Microbiol 88:123–131 [View Article][PubMed]
    [Google Scholar]
  20. Miller L., Berger T. 1985 Bacteria identification by gas chromatography of whole cell fatty acid. Hewlett Packard Application note 228-41, 1–8.
  21. Morandi S., Brasca M., Andrighetto C., Lombardi A., Lodi R. 2006; Technological and molecular characterisation of enterococci isolated from north–west Italian dairy products. Int Dairy J 16:867–875 [View Article]
    [Google Scholar]
  22. Morandi S., Brasca M., Lodi R. 2011; Technological, phenotypic and genotypic characterisation of wild lactic acid bacteria involved in the production of Bitto PDO Italian cheese. Dairy Sci Technol 91:341–359 [View Article]
    [Google Scholar]
  23. Naser S. M., Thompson F. L., Hoste B., Gevers D., Dawyndt P., Vancanneyt M., Swings J. 2005; Application of multilocus sequence analysis (MLSA) for rapid identification of Enterococcus species based on rpoA and pheS genes. Microbiology 151:2141–2150 [View Article][PubMed]
    [Google Scholar]
  24. Naser S. M., Vancanneyt M., Hoste B., Snauwaert C., Vandemeulebroecke K., Swings J. 2006; Reclassification of Enterococcus flavescens Pompei et al. 1992 as a later synonym of Enterococcus casseliflavus (ex Vaughan et al. 1979) Collins et al. 1984 and Enterococcus saccharominimus Vancanneyt et al. 2004 as a later synonym of Enterococcus italicus Fortina et al. 2004. Int J Syst Evol Microbiol 56:413–416 [View Article][PubMed]
    [Google Scholar]
  25. Schleifer K. H., Kilpper-Bälz R. 1984; Transfer of Streptococcus faecalis and Streptococcus faecium to the genus Enterococcus nom. rev. as Enterococcus faecalis comb. nov. and Enterococcus faecium comb. nov.. Int J Syst Bacteriol 34:31–34 [View Article]
    [Google Scholar]
  26. Stiles M. E., Holzapfel W. H. 1997; Lactic acid bacteria of foods and their current taxonomy. Int J Food Microbiol 36:1–29 [View Article][PubMed]
    [Google Scholar]
  27. Tanasupawat S., Sukontasing S., Lee J. S. 2008; Enterococcus thailandicus sp. nov., isolated from fermented sausage (‘mum’) in Thailand. Int J Syst Evol Microbiol 58:1630–1634 [View Article][PubMed]
    [Google Scholar]
  28. Tyrrell G. J., Turnbull L. A., Teixeira L. M., Lefebvre J., Carvalho M. G., Facklam R. R., Lovgren M. 2002; Enterococcus gilvus sp. nov. and Enterococcus pallens sp. nov. isolated from human clinical specimens. J Clin Microbiol 40:1140–1145 [View Article][PubMed]
    [Google Scholar]
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