Hybridization experiments were carried out between DNAs from more than 70 strains of Campylobacter spp. and related taxa and either 3H-labeled 23s rRNAs from reference strains belonging to Campylobacter fetus, Campylobacter concisus, Campylobacter sputorum, Campylobacter coli, and Campylobacter nitrofigilis, an unnamed Campylobacter sp. strain, and a Wolinella succinogenes strain or 3H-or 14C-labeled 23s rRNAs from 13 gram-negative reference strains. An immunotyping analysis of 130 antigens versus 34 antisera of campylobacters and related taxa was also performed. We found that all of the named campylobacters and related taxa belong to the same phylogenetic group, which we name rRNA superfamily VI and which is far removed from the gram-negative bacteria allocated to the five rRNA superfamilies sensu De Ley. There is a high degree of heterogeneity within this rRNA superfamily. Organisms belonging to rRNA superfamily VI should be reclassified in several genera. We propose that the emended genus Campylobacter should be limited to Campylo bacter fetus, Campylo bacter hy oin testinalis , Campylo bacter concisus, Campylo bacter m ucosalis , Campylobacter sputorum, Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and "Campylobacter upsaliensis. " Wolinella curva and Wolinella recta are transferred to the genus Campylobacter as Campylobacter curvus comb. nov. and Campylobacter rectus comb. nov., respectively. Bacteroides gracilis and Bacteroides ureolyticus are generically misnamed and are closely related to the genus Campylobacter. Campylobacter nitrofigilis, Campylobacter cryaerophila, and an unnamed Campylobacter sp. strain constitute a new genus, for which the name Arcobacter is proposed; this genus contains two species, Arcobacter nitrofigilis comb. nov. (type species) and Arcobacter cryaerophilus comb. nov. Wolinellu succinogenes so far is the only species of the genus Wolinella. The genus Helicobacter is also emended; Campylobacter cinaedi and Campylobacter fennelliae are included in this genus as Helicobacter cinaedi comb. nov. and Helicobacter fennelliae comb. nov., respectively. The genus "Flexispira," with "Flexispira rappini" as the only species, is closely related to the genus Helicobacter. The free-living, sulfur-reducing campylobacters do not belong to any of these genera; they probably constitute a distinct genus within rRNA superfamily VI.At present, the genus Campylobacter consists of 13 welldefined species (40). Recently, two additional species, Campylobacter pylori and Campylobacter mustelae, were included in the new genus Helicobacter as Helicobacter pylori and Helicobacter mustelae, respectively (20). The clinical significance of all of these organisms was reviewed recently by Penner (40). A study of the taxonomic structure of the genus Campylobacter in which partial 16s rRNA sequence analysis was used revealed that the Campylobacter species can be divided into three major rRNA homology groups (58 It was the aim of this study to include all known campylobacters and possible relatives in...
Two Gram-negative, non-motile, non-spore-forming, coccoid bacteria (strains CCM 4915 T and CCM 4916), isolated from clinical specimens of the common vole Microtus arvalis during an epizootic in the Czech Republic in 2001, were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA (rrs) and recA gene sequence similarities, both isolates were allocated to the genus Brucella. Affiliation to Brucella was confirmed by DNA-DNA hybridization studies. Both strains reacted equally with Brucella M-monospecific antiserum and were lysed by the bacteriophages Tb, Wb, F1 and F25. Biochemical profiling revealed a high degree of enzyme activity and metabolic capabilities not observed in other Brucella species. The omp2a and omp2b genes of isolates CCM 4915 T and CCM 4916 were indistinguishable. Whereas omp2a was identical to omp2a of brucellae from certain pinniped marine mammals, omp2b clustered with omp2b of terrestrial brucellae. Analysis of the bp26 gene downstream region identified strains CCM 4915 T and CCM 4916 as Brucella of terrestrial origin. Both strains harboured five to six copies of the insertion element IS711, displaying a unique banding pattern as determined by Southern blotting. In comparative multilocus VNTR (variable-number tandem-repeat) analysis (MLVA) with 296 different genotypes, the two isolates grouped together, but formed a separate Abbreviations: MLST, multilocus sequence typing; MLVA, multilocus VNTR (variable-number tandem-repeat) analysis; RTD, routine test dilution.The GenBank/EMBL/DDBJ accession numbers for the gene sequences omp22, omp25, omp25b, omp31 and omp31b of strain CCM 4915
A polyphasic taxonomic study that included DNA-DNA hybridizations, DNA base ratio determinations, 165 rDNA sequence analysis, whole-cell protein and fatty acid analyses, AFLP (amplified fragment length polymorphism) fingerprinting and an extensive biochemical characterization was performed on 10 strains provisionally identified as Alcaligenes faecalis-like bacteria. The six environmental and four human isolates belonged to the genus Ralstonia and were assigned to a new species for which the name Ralstonia gilardii sp. nov. is proposed. The type strain is LMG 5886l.
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...
Five strains of a hitherto unknown Actinomyces-like bacterium were isolated from human clinical sources, including blood cultures. Biochemical and chemotaxonomic characterization indicated that the strains were distinct from previously described Actinomyces and Arcanobacterium species. A comparative 16s rRNA gene sequence analysis demonstrated that the undescribed strains constitute a new subline within the ActinomycesArcanobacterium species complex. The closest known relative of the isolates was found to be Actinomyces suis, although a 16s rRNA sequence divergence value of approximately 6% clearly demonstrated that the unknown bacterium represents a distinct species. Based on the results of the present and earlier phylogenetic investigations, it is proposed that Actinomyces suis should be reclassified in a new genus, the genus Actinobaculum, as Actinobaculum suis comb, nov. In addition, a new species, Actinobaculum schualii, is proposed for the Actinomyces-like bacterium from human sources. The type strain of Actinobaculum schuulii is CCUG 27420.The genus Actinomyces is a heterogeneous group of anaerobic and facultatively anaerobic, asporogenous, gram-positive, non-acid-fast, rod-shaped organisms which generally show various degrees of branching (9, 10). Nearly all of the currently recognized Actinomyces species occur as inhabitants of mucosal surfaces of humans and other homoiothermic animals. Some Actinomyces species (e.g., Actinomyces bovis and Actinomyces israelii) have long been recognized as pathogens of humans and animals (9, 10). Several new Actinomyces species associated with disease in humans have been described in recent years (e.g., Actinomyces bernardiae [4], Actinomyces europae [5], Actinomyces neuii [3], Actinomyces turicensis [ 141, and Actinomyces radingue [ 14]), and Actinomyces-like organisms are attracting increasing interest as opportunistic pathogens. In this article we report the phenotypic and phylogenetic characterization of five strains of a hitherto unknown Actinomyceslike bacterium from human sources. Based on the taxonomic results presented below, a new species, Actinobaculum schaalii, is described.Five human clinical isolates (CCUG 19698, CCUG 29359B, CCUG 27420T, CCUG 32296, and CCUG 36567) were referred to the Culture Collection of the University of Goteborg (CCUG), Goteborg, Sweden, for identification. Strains CCUG 27420T and CCUG 36567 were isolated from human blood, whereas CCUG 19698, CCUG 29359B, and CCUG 32296 originated from human urine. All of the strains were cultured on 5% horse blood agar (Columbia base; Difco, Detroit, Mich.) at 37°C in a 5% CO, atmosphere. The isolates were biochemically characterized by using the API ZYM (enzymatic profiling), API coryne, API rapid ID 32 STREP, and API rapid ID 32A systems according to the instructions of the manufacturer
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