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Taxonomy of Antarctic Flavobacterium species: description of Flavobacterium gillisiae sp. nov., Flavobacterium tegetincola sp. nov., and Flavobacterium xanthum sp. nov., nom. rev. and reclassification of [Flavobacterium] salegens as Salegentibacter salegens gen. nov., comb. nov.

16S rRNA phylogenetic analysis of a number of yellow- and orange-pigmented strains isolated from a variety of Antarctic habitats including sea ice, lakewater and cyanobacterial mats indicated a close relationship to the genus Flavobacterium but distinct from known Flavobacterium species. Phenotypic properties, DNA G+C content and whole-cell fatty acid profiles of the Antarctic strains were consistent with those of the genus Flavobacterium. DNA-DNA hybridization analysis indicated the presence of two distinct and novel genospecies each isolated from a different Antarctic habitat. From polyphasic taxonomic data it is proposed that the two groups represent new species with the following proposed names: Flavobacterium gillisiae (ACAM 601T) and Flavobacterium tegetincola (ACAM 602T). In addition polyphasic analysis of the species '[Cytophaga] xantha' (Inoue and Komagata 1976), isolated from Antarctic mud, indicated it was a distinct member of the genus Flavobacterium and was thus revived as Flavobacterium xanthum. Phylogenetic and fatty acid analyses also indicate that the species [Flavobacterium] salegens (Dobson et al. 1993), from Organic Lake, Antarctica, is misclassified at the genus level. It is proposed that this species belongs to a new genus, Salegentibacter salegens gen. nov., comb. nov.

Antarctic Regions↗

Flavobacterium degerlachei sp. nov., Flavobacterium frigoris sp. nov. and Flavobacterium micromati sp. nov., novel psychrophilic bacteria isolated from microbial mats in Antarctic lakes.

Taxonomic studies were performed on 36 strains that were isolated from microbial mats in Antarctic lakes of the Vestfold Hills, the Larsemann Hills and the McMurdo Dry Valleys. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains are related to members of the genus Flavobacterium; sequence similarity values with their nearest phylogenetic neighbours ranged from 96.8 to 98.5%. Results of DNA-DNA hybridization and comparison of repetitive extragenic palindromic DNA-PCR fingerprinting patterns revealed that these strains are members of three distinct species. Genotypic results, together with phenotypic characteristics, allowed the differentiation of these species from related Flavobacterium species with validly published names. The isolates are Gram-negative, chemoheterotrophic, rod-shaped cells that are psychrophilic and moderately halotolerant; their DNA G+C contents range from 33.1 to 34.5 mol%. Their whole-cell fatty acid profiles are similar and include C(15:0), anteiso-C(15:0), iso-C(15:0), C(15:1)omega6c, iso-C(16:0), iso-C(16:0) 3-OH and summed feature 3 (which comprises iso-C(15:0) 2-OH, C(16:1)omega7c or both) as major fatty acid components. On the basis of these results, three novel species are proposed, namely Flavobacterium degerlachei sp. nov. (consisting of 14 strains, with LMG 21915T=DSM 15718T as the type strain), Flavobacterium micromati sp. nov. (consisting of three strains, with LMG 21919T=CIP 108161T as the type strain) and Flavobacterium frigoris sp. nov. (consisting of 19 strains, with LMG 21922T=DSM 15719T as the type strain).

Antarctic Regions↗

Chemical characterization of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf.

The cellular fatty acid, sphingolipid, and isoprenoid quinone compositions of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf were determined, using thin-layer, gas-liquid, and reverse-phase high-performance liquid chromatography. The fatty acid data showed that groups IIe, IIh, and IIf were similar to recognized Flavobacterium species by the presence of relatively large amounts of iso-branched hydroxy and nonhydroxy acids. Groups IIe and IIh were essentially identical in fatty acid composition but were distinguished from group IIf, F. breve, and F. odoratum on the basis of minor qualitative and quantitative differences. All strains tested contained menaquinone 6 as the major isoprenoid quinone, and all lacked sphingolipids. Overall, the chemical data suggest that groups IIe, IIh, and IIf are additional Flavobacterium species and are different from sphingobacteria, which contain sphingolipid and menaquinone 7 as the major quinone.

Chromatography, Gas↗

Flavobacterium daejeonense sp. nov. and Flavobacterium suncheonense sp. nov., isolated from greenhouse soils in Korea.

Two yellow-pigmented, Gram-negative, rod-shaped bacterial strains, GH1-10(T) and GH29-5(T), were isolated from greenhouse soils in Korea. 16S rRNA gene sequence analysis indicated that these strains were related to members of the genus Flavobacterium. Strain GH1-10(T) was most closely related to Flavobacterium psychrolimnae and Flavobacterium denitrificans, with sequence similarities of 95.9 and 95.2 %, respectively. Strain GH29-5(T) was most closely related to "Flavobacterium saliodium", F. denitrificans and Flavobacterium frigoris, with sequence similarities of 94.3, 92.5 and 92.5 %, respectively. The major cellular fatty acids of GH1-10(T) were iso-C(15 : 0), summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c) and iso-C(17 : 0) 3-OH, and those of GH29-5(T) were iso-C(15 : 0), iso-C(17 : 0) 3-OH, iso-C(15 : 1) G and iso-C(15 : 0) 3-OH. Both strains contained menaquinone with six isoprene units (MK-6) as the sole quinone. The DNA G+C contents of GH1-10(T) and GH29-5(T) were 35 and 39 mol%, respectively. Based on the phylogenetic and phenotypic data presented, it is concluded that the two bacteria represent two separate novel species of the genus Flavobacterium. The names proposed to accommodate these organisms are Flavobacterium daejeonense sp. nov., with type strain GH1-10(T) (=KACC 11422(T)=DSM 17708(T)), and Flavobacterium suncheonense sp. nov., with type strain GH29-5(T) (=KACC 11423(T)=DSM 17707(T)).

Bacterial Typing Techniques↗

Direct sequencing of the polymerase chain reaction-amplified 16S rRNA gene of Flavobacterium gondwanense sp. nov. and Flavobacterium salegens sp. nov., two new species from a hypersaline Antarctic lake.

Phenotypic data and phospholipid ester-linked fatty acid profiles indicate that pigmented bacterial strains isolated from a hypersaline Antarctic lake are members of the "flavobacterium-bacteroides" phylum and may represent new taxa. Nearly complete 16S rRNA sequences were obtained for representative strains by directly sequencing the polymerase chain reaction-amplified 16S rRNA gene. Sequence signatures confirmed that these organisms were members of the flavobacterium-bacteroides phylum. A phylogenetic analysis, in which the sequences of the Antarctic strains were compared with a large number of sequences available for members of the flavobacterium-bacteroides phylum, showed that the Antarctic strains were phylogenetically distinct. The new species cluster with a group of organisms that contains the type species of the genus Flavobacterium, Flavobacterium aquatile. Two new species are described, for which the names Flavobacterium gondwanense and Flavobacterium salegens are proposed; strains ACAM 44 (= DSM 5423) and ACAM 48 (= DSM 5424) are the type strains of F. gondwanense and F. salegens, respectively.

Antarctic Regions↗

Flavobacterium weaverense sp. nov. and Flavobacterium segetis sp. nov., novel psychrophiles isolated from the Antarctic.

Two psychrophilic flavobacteria, designated AT1042(T) and AT1048(T), were isolated from terrestrial samples from the Antarctic. Results of 16S rRNA gene sequence analyses indicated a close relationship of these isolates to Flavobacterium flevense (96.9 % similarity for strain AT1042(T)) and Flavobacterium psychrolimnae (97.0 % for strain AT1048(T)). Cells were non-motile and non-gliding. Flexirubin-type pigments were absent. Both isolates were psychrophilic, with an optimum and maximum growth temperature of about 15 and 20 degrees C, respectively. The major isoprenoid quinone, predominant cellular fatty acids and DNA G+C contents (35-37 mol%) were consistent with the placement of the Antarctic isolates in the genus Flavobacterium. Phylogeny based on 16S rRNA gene sequences and several phenotypic characteristics could be used to differentiate these isolates from recognized Flavobacterium species. Despite high 16S rRNA gene sequence similarity (98.9 %) between strains AT1042(T) and AT1048(T), they represented two distinct species as demonstrated by low genomic relatedness (34 %) and a number of differential phenotypic characters. The polyphasic data presented in this study indicated that the new isolates should be classified within two novel species in the genus Flavobacterium. The names Flavobacterium weaverense sp. nov. (type strain AT1042(T) = IMSNU 14048(T) = KCTC 12223(T) = JCM 12384(T)) and Flavobacterium segetis sp. nov. (type strain AT1048(T) = IMSNU 14050(T) = KCTC 12224(T) = JCM 12385(T)) are proposed for these Antarctic isolates.

Antarctic Regions↗

Flavobacterium xinjiangense sp. nov. and Flavobacterium omnivorum sp. nov., novel psychrophiles from the China No. 1 glacier.

Two novel psychrophilic bacterial strains (ZF-6(T) and ZF-8(T)) were isolated from the China No. 1 glacier. Polyphasic taxonomy using physiological and biochemical properties and phylogenetic analysis based on 18S rRNA gene sequences showed that the two isolates belonged to the genus Flavobacterium, and that they were distinct from each other and also from the known species of this genus. Strains ZF-6(T) and ZF-8(T) are Gram-negative and both have an optimal growth temperature of 11 degrees C. Strain ZF-6(T) is able to grow at 0-20 degrees C, the G + C content of its genomic DNA is 34.4 mol% and the major fatty acids of ZF-6(T) are C(16 : 1)omega7c (17.7%) and C15 : 1)omega6c (12.7%). Strain ZF-8(T) showed a strong ability to degrade organic macromolecules such as starch, CM-cellulose, pectin and chitin. Its DNA G + C content is 35.1 mol%, and the major fatty acids are C(16 : 1)omega7C (18.2%) and C(15 : 0) (9.9%). Phylogenetic analysis based on 16S rDNA sequences indicated that ZF-6(T) and ZF-8(T) belong to the genus Flavobacterium and represent two novel species. DNA-DNA hybridization also supported the status of the two new isolates. The names Flavobacterium xinjiangense sp. nov. (type strain, ZF-6(T) = AS 1.2749(T) = JCM 11314(T)) and Flavobacterium omnivorum sp. nov. (type strain, ZF-8(T) = AS 1.2747(T) = JCM 11313(T)) are proposed for the two new isolates.

Bacterial Typing Techniques↗

Flavobacterium fryxellicola sp. nov. and Flavobacterium psychrolimnae sp. nov., novel psychrophilic bacteria isolated from microbial mats in Antarctic lakes.

Taxonomic studies were performed on seven strains isolated from microbial mats in Antarctic lakes of the McMurdo Dry Valleys. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains are related to the genus Flavobacterium; sequence similarity values with their nearest phylogenetic neighbours ranged from 97.0 to 98.7 %. Results of DNA-DNA hybridization and comparison of repetitive extragenic palindromic DNA-PCR fingerprinting patterns revealed that these strains are members of two distinct species. Genotypic results, together with phenotypic characteristics, allowed these species to be differentiated from related recognized Flavobacterium species. The isolates are psychrophilic, Gram-negative, chemoheterotrophic, rod-shaped cells. Their whole-cell fatty acid profiles are similar and include C(15 : 0), C(15 : 0) anteiso, C(15 : 0) iso, C(15 : 1)omega6c, C(16 : 0) iso, C(16 : 0) iso 3-OH, C(16 : 1) iso and summed feature 3 (which comprises C(15 : 0) iso 2-OH and/or C(16 : 1)omega7c) as the major fatty acid components. On the basis of these results, two novel species are proposed: Flavobacterium fryxellicola sp. nov., consisting of three strains with LMG 22022(T) (=CIP 108325(T)) as the type strain; and Flavobacterium psychrolimnae sp. nov., consisting of four strains with LMG 22018(T) (=CIP 108326(T)) as the type strain. DNA G+C contents of F. fryxellicola and F. psychrolimnae are 35.2-35.9 and 33.8-34.5 mol%, respectively.

Antarctic Regions↗

Genotypic heterogeneity of Flavobacterium group IIb and Flavobacterium breve demonstrated by DNA-DNA hybridization.

DNA-DNA hybridization studies on 42 stains presumptively identified as members of Flavobacterium group IIb and Flavobacterium breve indicated pronounced genotypic heterogeneity within these taxa. Three large groups highly related to the type strains of F. gleum, F. indologenes and F. breve respectively, and eight small groups were found. The group containing the type strain of F. breve was phenotypically indistinguishable from another genomic group, and these two groups were significantly separated from the other flavobacteria studied. The other nine genomic groups, representing Flavobacterium group IIb, could not with certainty be differentiated from each other by phenotypic characteristics, and there is no evidence indicating that these genomic groups differ from each other with respect to pathogenicity or ecology. Thus, it is suggested that for the time being the name "Flavobacterium group IIb" rather than specific epithets continue to be used for these bacteria.

DNA, Bacterial↗

Occurrence of 2- and 3-hydroxy fatty acids in high concentrations in the extractable and bound lipids of Flavobacterium meningosepticum and Flavobacterium IIb.

The major hydroxy fatty acids of cellular lipids in Flavobacterium meningosepticum and Flavobacterium sp. King's group UUb were identified as 2-hydroxy 13-methyltetradecanoic, 3-hydroxy 13-methyltetradecanoic, 3-hydroxy palmitic, and 3-hydroxy 15-methylhexadecanoic acids using gas chromatography-mass spectrometry and GC-mass fragmentography. The concentration of these hydroxy fatty acids comprised up to 30-40% of the total extractable and 20-30% of the bound lipid fatty acids, respectively. From the stability for mild alkaline hydrolysis, 2-hydroxy fatty acids seemed to be attached with ester linkage, and 3-hydroxy fatty acids with amide linkage.

Chromatography, Gas↗

Identification of Flavobacterium columnare by a species-specific polymerase chain reaction and renaming of ATCC43622 strain to Flavobacterium johnsoniae.

Species-specific polymerase chain reaction (PCR) primers have been designed to identify the causative agent of columnaris disease, Flavobacterium columnare. The 16S rRNA gene sequences of F. columnare (eight sequences representing the different genotypes of the species) and related species (18 sequences) were aligned and compared to choose specific regions that are unique to F. columnare and do not have significant intraspecies variability. The species-specific regions in the 16S rRNA gene were used to design a pair of species-specific PCR primers, ColF and ColR. The PCR technique produced a specific amplicon of about 675 base pairs (bp) in 27 isolates of F. columnare and there was no amplification in the closely related species. The specificity of the amplified product was confirmed by digesting with HhaI. The PCR primers did not produce a 675 bp product with F. columnare ATCC43622 strain. This ATCC43622 strain was characterized by biochemical and ribotyping methods and renamed Flavobacterium johnsoniae. The American Type Culture Collection has confirmed these findings and made the change.

Bacterial Typing Techniques↗

Crossed immunoelectrophoretic analysis of Flavobacterium meningosepticum and allied Flavobacterium taxa.

A total of 55 different antigens were demonstrated in pooled lysate of six Flavobacterium meningosepticum strains by crossed immunoelectrophoresis against homologous rabbit antibody. The six strains used represented the two DNA relatedness groups within this species. All 52 strains of F. meningosepticum investigated were found to cross-react with 53 to 55 of these F. meningosepticum reference antigens. Using this F. meningosepticum reference system, cross-reactions between the F. meningosepticum reference antigen and antigens from representatives of other Flavobacterium taxa were also investigated. A positive correlation was demonstrated between the number of cross-reacting antigens and the taxonomic relatedness of the investigated taxa as determined by other classification methods, including DNA-DNA hybridization. The crossed immunoelectrophoretic techniques were not useful in differentiating the two main DNA groups of F. meningosepticum.

Animals↗

Cellular fatty acids of Flavobacterium meningosepticum and Flavobacterium species group IIb.

The cellular fatty acid profiles of Flavobacterium meningosepticum and Flavobacterium species group IIb were markedly different from those of related bacteria. The profiles were characterized by the presence of 13-methyl-tetradecanoate and three uncommon acids: 2-hydroxy-13-methyl-tetradecanoate, 15-methyl-hexadecanoate, and 3-hydroxy-15-methyl-hexadecanoate.

Chromatography, Gas↗