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The taxonomic status of two bacterial strains isolated from human blood was characterized using a polyphasic approach. Chemotaxonomic investigations revealed the presence of cell-wall chemotype IV, short-chain mycolic acids that co-migrated with those extracted from members of the genus Williamsia and that produce C 16 : 0 and C 18 : 0 fatty acids on pyrolysis GC, and dihydrogenated menaquinone with nine isoprene units as the predominant isoprenologue. The generic assignment was confirmed by 16S rRNA gene sequencing. Comparative analysis of the 16S rRNA gene sequence showed that these isolates constitute a distinct phyletic line within the genus Williamsia, displaying 96?2 and 97?2 % sequence similarities to Williamsia muralis and Williamsia maris, respectively. The two isolates could be distinguished from the type strains of the latter species on the basis of several phenotypic traits. The genotypic and phenotypic data show that the strains merit classification as a novel species of Williamsia, for which the name Williamsia deligens sp. nov. is proposed, with type strain IMMIB RIV-956The genus Williamsia was proposed by Kämpfer et al. (1999) to accommodate actinomycetes with atypical cell morphology as revealed under electron microscopy that are unable to grow at 5 or at 45 uC and possess mycolic acids with carbon chain lengths of 50 to 56. Based on its mycolic acids, it seems that Williamsia takes an intermediate position between Rhodococcus (mycolic acid chain lengths of 34-45) and Gordonia (mycolic acid chain lengths of 54-66) (Kämpfer et al., 1999). The genus Williamsia currently comprises two recognized species, Williamsia muralis isolated from indoor building material of a children's day-care centre in Finland (Kämpfer et al., 1999) and Williamsia maris isolated from deep sediments of the Sea of Japan (Stach et al., 2004). In this paper we describe two bacterial strains which were isolated from human blood. Based on phylogenetic and phenotypic data it is proposed that these strains (designated IMMIB RIV-956 T and IMMIB RIV956Fl) are similar and should be classified as representing a novel species of the genus Williamsia. Isolates IMMIB RIV-956T and IMMIB RIV-956Fl were isolated from human blood. The type strains of W. maris (DSM 44693 T ) and W. muralis (DSM 44343 T ) were received from the DSMZ. All strains were cultured on Columbia agar supplemented with 5 % sheep blood agar and brain heart infusion (BHI) agar to determine their morphological characteristics. Production of pigments was determined by growing the strains at 27 u C for 7 days, and observations were made at 24 h intervals. Air-dried smears at 24, 48 and 72 h intervals were stained by using the Gram's method in order to determine the Gram reaction and cell morphology.

A bacterial isolate obtained from soil from a municipal landfill site in India was characterized using a polyphasic taxonomic approach. The colonies of the isolate were found to be yellow and highly mucoid. Comparative analysis of the 16S rRNA gene sequence showed that this isolate constitutes a distinct phyletic line within the genus Lysobacter, displaying >3 % sequence divergence with respect to recognized Lysobacter species. The generic assignment was confirmed by chemotaxonomic data, which revealed the presence of a fatty acid profile characteristic of members of the genus Lysobacter and consisting of saturated, unsaturated, straight-chain and branched-chain fatty acids as well as iso-C 11 : 0 3-OH as hydroxylated fatty acid, and the presence of an ubiquinone with eight isoprene units (Q-8) as the predominant respiratory quinone. The genotypic and phenotypic data show that strain IMMIB APB-9T merits classification as representing a novel species of the genus Lysobacter, for which the name Lysobacter defluvii sp. nov. is proposed. The type strain is IMMIB APB-9 T (=CCUG 53152 T =DSM 18482 T ).

We describe a new genus of mesophilic actinomycetes, for which we propose the name Lentzea. The strains of this genus form abundant aerial hyphae that fragment into rod-shaped elements. Whole-cell hydrolysates contain the meso isomer of diaminopimelic acid and no characteristic sugar (wall chemotype 111). The phospholipid pattern type is type PI1 (phosphatidylethanolamine is the characteristic phospholipid); the major menaquinone is MK-9. The fatty acid profile comprises saturated, unsaturated, and branched-chain fatty acids of the is0 and anteiso types in addition to tuberculostearic acid (lOMe-C,,:,). A 16s ribosomal DNA sequence analysis revealed that the genus Lentzea is phylogenically related to the genera Actinosynnema, Sacchurothrix, and Kutzneria. The type species of this genus is Lentzea albidocupillatu sp. nov.; the type strain of this species is strain IMMIB D-958 (= DSM 44073).

Phospholipid patterns of 15 representative strains of the genus Amycolatopsis were recorded by twodimensional thin-layer chromatography. The structure analysis of the isolated phospholipids was verified by fast atom bombardment-mass spectroscopy. The positive-and negative-ion spectra of the partially purified phospholipid fractions qualitatively reflect their distinctive composition. All strains contained diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylinositol. Two different types of phosphatidylethanolamine and phosphatidylmethylethanolamine were detected, viz., compounds with or without hydroxy fatty acids. These phospholipid patterns underline the integrity of the genus. Fast atom bombardment-mass spectrometry analysis of phospholipid patterns may serve as an aid for differentiation of bacterial species.Valuable taxonomic information for actinomycete systematics has been derived from two-dimensional thin-layer chromatographic (TLC) analyses of polar lipids (16,(25)(26)(27). Phospholipids are the most common actinomycete polar lipids, and they have been proven to be of value as taxonomic markers (10, 15, 16, 18, 21, 22, 24, 32). The phospholipids most frequently encountered in actinomycete wall envelopes include phosphatidylcholine (PC), phosphatidyl- glycerol (PG), phosphatidylethanolamine (PE), and phosphatidylinositol (PI). Less commonly detected polar lipids include phosphatidylmethylethanolamine (PME).Lechevalier et al. (16, 18) divided the phospholipids extracted from aerobic actinomycetes into five groups on the basis of the distribution of certain nitrogenous phospholipids. Members of the family Pseudonocardiaceae were recovered in two of these groups. Among them, Amycolafopsk, Kibdelosporangium, and Saccharomonospora strains have a type I1 phospholipid pattern; that means that they contain PE but not phosphatidylcholine.Characterization and identification of microorganisms constitute a major goal of diagnostic microbiology. Many systems and methods have been developed for this task, but in recent years increasing emphasis has been placed on the need to reduce the time involved and to introduce automated systems into diagnostic microbiology. One of the methods likely to enable rapid characterization of microorganisms is fast atom bombardment (FAB)-mass spectrometry (MS isolated from different species of the genus Amycolatopsis were subjected to negative-and positive-ion FAB as well as FAB-tandem MS to verify their structural assignments as already identified by two-dimensional TLC and to evaluate the usefulness of these methods for phospholipid analysis from bacterial sources. MATERIALS AND METHODSStrains and culture conditions. Stock cultures were taken from the culture collection of the Institute for Medical Microbiology and Immunology of the University of Bonn as listed in Table 1. The strains were maintained on brain heart infusion agar (Difco) at 4°C. The strains were grown in brain heart infusion broth (Difco) under agitation on a rotary shaker (150 rpm) for 7 days. At maximum grow...

A bacterial isolate from a sample of oil-contaminated soil was characterized using a polyphasic taxonomic approach. Comparative analysis of the 16S rRNA gene sequence showed that this isolate constituted a distinct phyletic line within the genus Pseudoxanthomonas, displaying .3.7 % sequence divergence with respect to recognised Pseudoxanthomonas species. The genus assignment was confirmed by a chemotaxonomic analysis, which revealed the presence of a fatty acid profile characteristic of members of the genus Pseudoxanthomonas (straight-chain saturated, unsaturated and branched-chain fatty acids of the iso/anteiso type and 3-hydroxylated fatty acids) and the presence of a ubiquinone with eight isoprene units (Q-8) as the predominant respiratory quinone. The novel isolate was distinguishable from other members of the genus Pseudoxanthomonas on the basis of a combination of phenotypic properties. The genotypic and phenotypic data show that the strain represents a novel species of the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas spadix sp. nov. is proposed. The type strain is IMMIB AFH-5 T (5DSM 18855 T 5CCUG 53828 T ).

Two bacterial isolates, strains IMMIB RIV-085T and IMMIB RIV-095, isolated from a blood-sucking bug of the genus Triatoma, were characterized by phenotypic and molecular taxonomic methods. Chemotaxonomic investigations revealed the presence of cell-wall chemotype IV and mycolic acids consistent with the genus Rhodococcus. Comparative 16S rRNA gene sequencing showed that the two isolates are genealogically highly related (100 % sequence similarity) and constitute a new subline within the genus Rhodococcus, with Rhodococcus corynebacteroides and Rhodococcus rhodnii as their nearest phylogenetic neighbours (98·4 and 98·3 % sequence similarity, respectively). However, DNA–DNA hybridization experiments demonstrated unambiguously that the isolates are genealogically distinct from R. corynebacteroides and R. rhodnii (32 and 43 % relatedness, respectively). The isolates could be distinguished from their phylogenetic relatives and other members of the genus Rhodococcus by means of biochemical tests. On the basis of both phenotypic and phylogenetic evidence, it is proposed that these isolates be classified as a novel species, Rhodococcus triatomae sp. nov. The type strain is strain IMMIB RIV-085T (=CCUG 50202T=DSM 44892T).