Downloaded from www.microbiologyresearch.org by IP: 54.162.210.129 On: Sat, 23 Jan 2016 14:17:03 Burkholderia latens sp. nov., Burkholderia diffusa sp. nov., Burkholderia arboris sp. nov., Burkholderia seminalis sp. nov. and Burkholderia metallica sp. nov., novel species within the Burkholderia cepacia complex Elke Vanlaere, 1 John J. LiPuma, 2 Adam Baldwin, 3 Deborah Henry, 4 Evie De Brandt, 1 Eshwar Mahenthiralingam, 5 David Speert, 4 Chris Dowson 3 and Peter Vandamme 1 Correspondence Peter Vandamme Peter.Vandamme@UGent.be 1 Laboratorium voor Microbiologie, Universiteit Gent, K. L. Ledeganckstraat 35, B-9000 Gent, Belgium 2 Department of Pediatrics, University of Michigan, 1150 W. Medical Centre Drive, 8323 MRSB III, Box 0646, Ann Arbor, MI 48109-0646, USA 3 Department of Biological Sciences, Warwick University, Coventry CV4 7AL, UK 4 Department of Pediatrics, University of British Columbia, Vancouver, BC, Canada 5 Cardiff School of Biosciences, Cardiff University, Cardiff CF10 3TL, UK The taxonomic position of five recA gene clusters of Burkholderia cepacia complex (Bcc) isolates was determined using a polyphasic taxonomic approach. The levels of 16S rRNA and recA 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 recA gene sequence analysis, MLST and BOX-PCR profiling and by recA RFLP analysis. For diagnostic laboratories, recA gene sequence analysis offers the best combination of accuracy and simplicity. Based on these results, we propose five novel Bcc species, Burkholderia latens sp. nov. (type strain FIRENZE 3 T 5LMG 24064 T 5CCUG 54555 T ), Burkholderia diffusa sp. nov. (type strain AU1075 T 5LMG 24065 T 5CCUG 54558 T ), Burkholderia arboris sp. nov. (type strain ES0263A T 5LMG 24066 T 5CCUG 54561 T ), Burkholderia seminalis sp. nov. (type strain AU0475 T 5LMG 24067 T 5CCUG 54564 T ) and Burkholderia metallica sp. nov. (type strain AU0553 T 5LMG 24068 T 5CCUG 54567 T ). In the present study, we also demonstrate that Burkholderia ubonensis should be considered a member of the Bcc. INTRODUCTION The Burkholderia cepacia complex (Bcc) is a diverse group of bacteria (Coenye et al., 2001c; Vandamme et al., 1997) which are ubiquitous in nature (Coenye & Vandamme, 2003). Some Bcc strains are recognized for their beneficial properties as plant-growth promoters, bioremediators and biocontrol strains (Parke & Gurian-Sherman, 2001), whereas others are notorious as pathogens causing infections in plants and immunocompromised individuals such as cystic fibrosis patients (Coenye & Vandamme, 2003; Mahenthiralingam et al., 2005). The Bcc currently comprises nine species, which share a high degree of 16S rRNA gene (98–100 %) and recA Abbreviations: Bcc, Burkholderia cepacia complex; MLST, multilocus sequence typing; RFLP, restriction fragment length polymorphism. The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains LMG 24064 T , LMG 24065 T , LMG 24066 T , LMG 24067 T and LMG 24068 T are AM747628–AM747632, and those of the recA sequences of strains LMG 24275, LMG 24066 T , R-24202, LMG 19587, LMG 24271, LMG 24265, R-22599, R-23401, LMG 24067 T , LMG 24065 T , LMG 24270, LMG 19239 and LMG 24064 T are AM748094–AM748105 and AM922300. Results of DNA–DNA hybridization experiments, HaeIII/MnlI recA gene RFLP patterns and a dendrogram derived from UPGMA linkage of correlation coefficients between BOX-PCR fingerprints are available as supplementary material with the online version of this paper. International Journal of Systematic and Evolutionary Microbiology (2008), 58, 1580–1590 DOI 10.1099/ijs.0.65634-0 1580 65634 G 2008 IUMS Printed in Great Britain