The taxonomic position of five gene clusters of complex (Bcc) isolates was determined using a polyphasic taxonomic approach. The levels of 16S rRNA and gene sequence similarity, multilocus sequence typing (MLST) data and the intermediate DNA–DNA binding values demonstrated that these five clusters represented five novel species within the Bcc. Biochemical identification of these species is difficult, as is the case for most Bcc species. However, identification of these novel species can be accomplished through gene sequence analysis, MLST and BOX-PCR profiling and by RFLP analysis. For diagnostic laboratories, gene sequence analysis offers the best combination of accuracy and simplicity. Based on these results, we propose five novel Bcc species, sp. nov. (type strain FIRENZE 3 =LMG 24064 =CCUG 54555), sp. nov. (type strain AU1075 =LMG 24065 =CCUG 54558), sp. nov. (type strain ES0263A =LMG 24066 =CCUG 54561), sp. nov. (type strain AU0475 =LMG 24067 =CCUG 54564) and sp. nov. (type strain AU0553 =LMG 24068 =CCUG 54567). In the present study, we also demonstrate that should be considered a member of the Bcc.


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  1. Baldwin, A., Mahenthiralingam, E., Thickett, K. M., Honeybourne, D., Maiden, M. C., Govan, J. R., Speert, D. P., LiPuma, J. J., Vandamme, P. & Dowson, C. G.(2005). Multilocus sequence typing scheme that provides both species and strain differentiation for the Burkholderia cepacia complex. J Clin Microbiol 43, 4665–4673.[CrossRef] [Google Scholar]
  2. Chen, J. S., Witzmann, K. A., Spilker, T., Fink, R. J. & LiPuma, J. J.(2001). Endemicity and inter-city spread of Burkholderia cepacia genomovar III in cystic fibrosis. J Pediatr 139, 643–649.[CrossRef] [Google Scholar]
  3. Coenye, T. & Vandamme, P.(2003). Diversity and significance of Burkholderia species occupying diverse ecological niches. Environ Microbiol 5, 719–729.[CrossRef] [Google Scholar]
  4. Coenye, T., Laevens, S., Willems, A., Ohlén, M., Hannant, W., Govan, J. R., Gillis, M., Falsen, E. & Vandamme, P.(2001a).Burkholderia fungorum sp. nov. and Burkholderia caledonica sp. nov., two new species isolated from the environment, animals and human clinical samples. Int J Syst Evol Microbiol 51, 1099–1107.[CrossRef] [Google Scholar]
  5. Coenye, T., Mahenthiralingam, E., Henry, D., LiPuma, J. J., Laevens, S., Gillis, M., Speert, D. P. & Vandamme, P.(2001b).Burkholderia ambifaria sp. nov., a novel member of the Burkholderia cepacia complex including biocontrol and cystic fibrosis-related isolates. Int J Syst Evol Microbiol 51, 1481–1490. [Google Scholar]
  6. Coenye, T., Vandamme, P., Govan, J. R. & LiPuma, J. J.(2001c). Taxonomy and identification of the Burkholderia cepacia complex. J Clin Microbiol 39, 3427–3436.[CrossRef] [Google Scholar]
  7. Coenye, T., Spilker, T., Martin, A. & LiPuma, J. J.(2002). Comparative assessment of genotyping methods for epidemiologic study of Burkholderia cepacia genomovar III. J Clin Microbiol 40, 3300–3307.[CrossRef] [Google Scholar]
  8. Coenye, T., Spilker, T., Van Schoor, A., LiPuma, J. J. & Vandamme, P.(2004). Recovery of Burkholderia cenocepacia strain PHDC from cystic fibrosis patients in Europe. Thorax 59, 952–954.[CrossRef] [Google Scholar]
  9. Ezaki, T., Hashimoto, Y. & Yabuuchi, E.(1989). 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 39, 224–229.[CrossRef] [Google Scholar]
  10. Gillis, M., Van Van, T., Bardin, R., Goor, M., Hebbar, P., Willems, A., Segers, P., Kersters, K., Heulin, T. & Fernandez, M. P.(1995). Polyphasic taxonomy in the genus Burkholderia leading to an emended description of the genus and proposition of Burkholderia vietnamiensis sp. nov. for N2-fixing isolates from rice in Vietnam. Int J Syst Bacteriol 45, 274–289.[CrossRef] [Google Scholar]
  11. Goris, J., Suzuki, K., De Vos, P., Nakase, T. & Kersters, K.(1998). Evaluation of a microplate DNA-DNA hybridization method compared with the initial renaturation method. Can J Microbiol 44, 1148–1153.[CrossRef] [Google Scholar]
  12. Goris, J., Dejonghe, W., Falsen, E., De Clerck, E., Geeraerts, B., Willems, A., Top, E. M., Vandamme, P. & De Vos, P.(2002). Diversity of transconjugants that acquired plasmid pJP4 or pEMT1 after inoculation of a donor strain in the A- and B-horizon of an agricultural soil and description of Burkholderia hospita sp. nov. and Burkholderia terricola sp. nov. Syst Appl Microbiol 25, 340–352.[CrossRef] [Google Scholar]
  13. Haubold, B. & Hudson, R. R.(2000).lian 3.0: detecting linkage disequilibrium in multilocus data. Bioinformatics 16, 847–848.[CrossRef] [Google Scholar]
  14. Henry, D. A., Campbell, M. E., LiPuma, J. J. & Speert, D. P.(1997). Identification of Burkholderia cepacia isolates from patients with cystic fibrosis and use of a simple new selective medium. J Clin Microbiol 35, 614–619. [Google Scholar]
  15. Henry, D. A., Mahenthiralingam, E., Vandamme, P., Coenye, T. & Speert, D. P.(2001). Phenotypic methods for determining genomovar status of the Burkholderia cepacia complex. J Clin Microbiol 39, 1073–1078.[CrossRef] [Google Scholar]
  16. Jolley, K. A., Chan, M.-S. & Maiden, M. C.(2004). mlstdbNet – distributed multi-locus sequence typing (MLST) databases. BMC Bioinformatics 5, 86[CrossRef] [Google Scholar]
  17. Kumar, S., Tamura, K. & Nei, M.(2004).mega3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5, 150–163.[CrossRef] [Google Scholar]
  18. LiPuma, J. J., Spilker, T., Coenye, T. & Gonzalez, C. F.(2002). An epidemic Burkholderia cepacia complex strain identified in soil. Lancet 359, 2002–2003.[CrossRef] [Google Scholar]
  19. LiPuma, J. J., Currie, B. J., Lum, G. D. & Vandamme, P. A. R.(2007).Burkholderia, Stenotrophomonas, Ralstonia, Cupriavidus, Pandoraea, Brevundimonas, Comamonas, Delftia, and Acidovorax. In Manual of Clinical Microbiology, 9th edn, vol. 1, pp. 749–769. Edited by P. R. Murray, E. J. Baron, J. H. Jorgensen, M. L. Landry & M. A. Pfaller. Washington, DC: American Society for Microbiology.
  20. Mahenthiralingam, E., Coenye, T., Chung, J. W., Speert, D. P., Govan, J. R., Taylor, P. & Vandamme, P.(2000a). Diagnostically and experimentally useful panel of strains from the Burkholderia cepacia complex. J Clin Microbiol 38, 910–913. [Google Scholar]
  21. Mahenthiralingam, E., Bischof, J., Byrne, S. K., Radomski, C., Davies, J. E., Av-Gay, Y. & Vandamme, P.(2000b). DNA-based diagnostic approaches for identification of Burkholderia cepacia complex, Burkholderia vietnamiensis, Burkholderia multivorans, Burkholderia stabilis, and Burkholderia cepacia genomovars I and III. J Clin Microbiol 38, 3165–3173. [Google Scholar]
  22. Mahenthiralingam, E., Baldwin, A. & Vandamme, P.(2002).Burkholderia cepacia complex infection in patients with cystic fibrosis. J Med Microbiol 51, 533–538. [Google Scholar]
  23. Mahenthiralingam, E., Urban, T. A. & Goldberg, J. B.(2005). The multifarious, multireplicon Burkholderia cepacia complex. Nat Rev Microbiol 3, 144–156.[CrossRef] [Google Scholar]
  24. Marmur, J.(1961). A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3, 208–218.[CrossRef] [Google Scholar]
  25. McDowell, A., Mahenthiralingam, E., Moore, J. E., Dunbar, K. E., Webb, A. K., Dodd, M. E., Martin, S. L., Millar, B. C., Scott, C. J. & other authors(2001). PCR-based detection and identification of Burkholderia cepacia complex pathogens in sputum from cystic fibrosis patients. J Clin Microbiol 39, 4247–4255.[CrossRef] [Google Scholar]
  26. Mesbah, M. & Whitman, W. B.(1989). Measurement of deoxyguanosine/thymidine ratios in complex mixtures by high-performance liquid chromatography for determination of the mole percentage guanine + cytosine of DNA. J Chromatogr 479, 297–306.[CrossRef] [Google Scholar]
  27. Parke, J. L. & Gurian-Sherman, D.(2001). Diversity of the Burkholderia cepacia complex and implications for risk assessment of biological control strains. Annu Rev Phytopathol 39, 225–258.[CrossRef] [Google Scholar]
  28. Payne, G. W., Vandamme, P., Morgan, S. H., LiPuma, J. J., Coenye, T., Weightman, A. J., Jones, T. H. & Mahenthiralingam, E.(2005). Development of a recA gene-based identification approach for the entire Burkholderia genus. Appl Environ Microbiol 71, 3917–3927.[CrossRef] [Google Scholar]
  29. Pitcher, D. G., Saunders, N. A. & Owen, R. J.(1989). Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett Appl Microbiol 8, 151–156.[CrossRef] [Google Scholar]
  30. Rademaker, J. L., Hoste, B., Louws, F. J., Kersters, K., Swings, J., Vauterin, L., Vauterin, P. & de Bruijn, F. J.(2000). Comparison of AFLP and rep-PCR genomic fingerprinting with DNA–DNA homology studies: Xanthomonas as a model system. Int J Syst Evol Microbiol 50, 665–677.[CrossRef] [Google Scholar]
  31. Storms, V., Van den Vreken, N., Coenye, T., Mahenthiralingam, E., LiPuma, J. J., Gillis, M. & Vandamme, P.(2004). Polyphasic characterisation of Burkholderia cepacia-like isolates leading to the emended description of Burkholderia pyrrocinia. Syst Appl Microbiol 27, 517–526.[CrossRef] [Google Scholar]
  32. Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. & Higgins, D. G.(1997). The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25, 4876–4882.[CrossRef] [Google Scholar]
  33. Vandamme, P., Holmes, B., Vancanneyt, M., Coenye, T., Hoste, B., Coopman, R., Revets, H., Lauwers, S., Gillis, M. & other authors(1997). Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderia multivorans sp. nov. Int J Syst Bacteriol 47, 1188–1200.[CrossRef] [Google Scholar]
  34. Vandamme, P., Mahenthiralingam, E., Holmes, B., Coenye, T., Hoste, B., De Vos, P., Henry, D. & Speert, D. P.(2000). Identification and population structure of Burkholderia stabilis sp. nov. (formerly Burkholderia cepacia genomovar IV). J Clin Microbiol 38, 1042–1047. [Google Scholar]
  35. Vandamme, P., Henry, D., Coenye, T., Nzula, S., Vancanneyt, M., LiPuma, J. J., Speert, D. P., Govan, J. R. & Mahenthiralingam, E.(2002).Burkholderia anthina sp. nov. and Burkholderia pyrrocinia, two additional Burkholderia cepacia complex bacteria, may confound results of new molecular diagnostic tools. FEMS Immunol Med Microbiol 33, 143–149.[CrossRef] [Google Scholar]
  36. Vandamme, P., Holmes, B., Coenye, T., Goris, J., Mahenthiralingam, E., LiPuma, J. J. & Govan, J. R.(2003).Burkholderia cenocepacia sp. nov. – a new twist to an old story. Res Microbiol 154, 91–96.[CrossRef] [Google Scholar]
  37. Vandamme, P. A. R., Govan, J. R. W. & LiPuma, J. J.(2007). Diversity and role of Burkholderia spp. In Burkholderia Molecular Microbiology and Genomics, vol. 1, pp. 1–28. Edited by T. Coenye & P. Vandamme. Wymondham, UK: Horizon Bioscience.
  38. Vanlaere, E., Coenye, T., Samyn, E., Van Den Plas, C., Govan, J., De Baets, F., De Boeck, K., Knoop, C. & Vandamme, P.(2005). A novel strategy for the isolation and identification of environmental Burkholderia cepacia complex bacteria. FEMS Microbiol Lett 249, 303–307.[CrossRef] [Google Scholar]
  39. Vermis, K., Coenye, T., LiPuma, J. J., Mahenthiralingam, E., Nelis, H. J. & Vandamme, P.(2004). Proposal to accommodate Burkholderia cepacia genomovar VI as Burkholderia dolosa sp. nov. Int J Syst Evol Microbiol 54, 689–691.[CrossRef] [Google Scholar]
  40. Yabuuchi, E., Kawamura, Y., Ezaki, T., Ikedo, M., Dejsirilert, S., Fujiwara, N., Naka, T. & Kobayashi, K.(2000).Burkholderia uboniae sp. nov., l-arabinose-assimilating but different from Burkholderia thailandensis and Burkholderia vietnamiensis. Microbiol Immunol 44, 307–317.[CrossRef] [Google Scholar]

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vol. , part 7, pp. 1580 - 1590

DNA base ratios and DNA–DNA binding values of all strains examined.

III/ I gene RFLP patterns observed in clusters 1–5.

Dendrogram derived from UPGMA linkage of correlation coefficients between BOX-PCR fingerprint patterns from Bcc isolates.

[PDF file of Supplementary Table S1 and Figs S1 and S2](313 KB)

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