A polyphasic taxonomic study was performed on a group of isolates tentatively identified as Burkholderia cepacia, Ralstonia pickettii or Ralstonia paucula (formerly known as CDC group IVc-2). The isolates were mainly cultured from sputum of cystic fibrosis patients or from soil. SDS-PAGE of whole-cell proteins and AFLP fingerprinting distinguished at least five different species, and this was confirmed by DNA-DNA hybridizations. 16S rDNA sequence analysis of representative strains indicated that these organisms belong to the β-subclass of the Proteobacteria, with the genera INTRODUCTIONThe majority of life-threatening bacterial lung infections in cystic fibrosis (CF) patients are caused by a limited spectrum of bacterial pathogens which include Pseudomonas aeruginosa, Staphylococcus aureus and Burkholderia cepacia (Gilligan, 1991 ;Govan & Deretic, 1996). Other organisms which can be found regularly in respiratory tract specimens of CF patients include Haemophilus influenzae, Stenotro- phomonas maltophilia, certain fungi (e.g. Aspergillus spp.) and viruses (e.g. respiratory syncytial virus) (Gilligan, 1991). In the absence of epidemic spread, B. cepacia only infects a small proportion of CF patients, but it has a major impact on both morbidity and mortality of infected patients and the medical and psychosocial implications of B. cepacia colonization and segregation of colonized and non-colonized patients as a means of infection control are enormous (LiPuma, 1998). Accurate identification of B. cepacia is crucial in patient management and infection control practices, but may be problematic, and misidentification is relatively common (LiPuma, 1998 Vandamme et al., 1999). This report describes the polyphasic taxonomic study which was used for the characterization of these and similar isolates from the environment and human clinical specimens. The genotypic and phenotypic characteristics warranted the classification of the isolates in a novel genus, for which we propose the name Pandoraea. The genus Pandoraea contains five species : Pandoraea apista sp. nov. (the type species), Pandoraea pulmonicola sp. nov., Pandoraea pnomenusa sp. nov., Pandoraea sputorum sp. nov. and Pandoraea norimbergensis (Wittke et al. 1997) comb. nov. (basonym Burkholderia norimbergensis). The name Burkholderia norimbergensis was proposed by Wittke et al. (1997) for a single bacterial isolate capable of hetrotrophic sulfur oxidation, which was isolated from an oxic water layer above a sulfide-containing sediment. METHODSBacterial strains and growth conditions. Strains used in this study are listed in Table 1. All strains were grown aerobically on Trypticase Soy Agar (BBL) at 37 mC unless otherwise indicated.Phenotypic characterization. Classical phenotypic tests were performed as described previously (Vandamme et al., 1993). API ZYM and API20 NE tests were performed according to the recommendations of the manufacturer (bioMe! rieux). Flagella staining was performed using a solution of tannic acid (2 g)\phenol (5 %, w\v)\KAl(SO % ) # (satura...

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Eleven strains of a hitherto undescribed Gram-positive, catalase-negative, facultatively anaerobic rod-shaped bacterium from human sources and medical care products were characterized by phenotypic and molecular taxonomic methods. The phenotypic properties of the bacterium were consistent with its assignment to the genus Lactobacillus but it was readily distinguished from all currently described species of this genus by its biochemical characteristics and by SDS-PAGE analysis of its cellular proteins. Comparative 16s rRNA gene sequence analysis demonstrated that the unknown bacterium was a member of rRNA group I Lactobacillus which includes Lactobacillus delbrueckii, the type species of the genus, and close relatives. Lactobacillus gasseri and Lactobacillus johnsonii were the nearest phylogenetic relatives of the unknown bacterium, but 165 rRNA sequence divergence values of ~4 % clearly showed that it represents a distinct species. Based on both phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium should be classified in the genus Lactobacillus, as Lactobacillus hers sp. nov. The type strain of Lactobacillus hers is CCUG 28746T.

Phenotypic and phylogenetic studies were performed on an unknown Grampositive catalase-negative coccus isolated from a common otter (Lufra lutra).Comparative 16s rRNA gene sequencing demonstrated that the unknown bacterium represents a new subline within the genus Vagococcus, close to, but distinct from, Vagococcus fluvialis and Vagococcus salmoninarum. The unknown bacterium was readily distinguished from the two currently recognized Vagococcus species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium be classified as a new species, Vagococcus lutrae, the type strain of which is CCUG 39187T.

Phenotypic and phylogenetic studies were performed on two strains of a hitherto undescribed Aerococcus-like organism isolated from the human vagina. Comparative 16s rRNA gene sequencing studies demonstrated that the unknown strains constitute a new subline within the genus Aerococcus. The unknown bacterium was readily distinguished from the two currently recognized Aerococcus species, Aerococcus viridans and Aerococcus urinae, by biochemical tests and electrophoretic analysis of whole-cell proteins. On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Aerococcus christensenii sp. nov. The type strain of A. christensenii is CCUG 2883IT.