- Volume 60, Issue 2, 2010

A novel Gram-stain-positive, mycelium-forming actinobacterium (05-Be-005T) isolated from the wall of an indoor environment was studied for its taxonomic position. The isolated strain formed a substrate mycelium that fragmented into rod-shaped cells and showed an aerial mycelium on medium M79. On the basis of 16S rRNA gene sequence similarity studies, strain 05-Be-005T was shown to belong to the genus Prauserella, closely related to Prauserella rugosa DSM 43194T (96.6 % similarity), Prauserella alba YIM 90005T (95.9 %) and Prauserella halophila YIM 90001T (95.4 %). The predominant menaquinone was MK-9(H4); whole-cell hydrolysates contained meso-diaminopimelic acid as the diagnostic diamino acid of the cell wall and arabinose and galactose as the main sugars. Mycolic acids were absent. The polar lipid profile consisted of the lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylserine and an unknown phospholipid. Major fatty acids C16 : 0 iso, C16 : 0, C17 : 1 ω8c and C17 : 1 ω6c supported the affiliation of strain 05-Be-005T to the genus Prauserella. The results of physiological and biochemical tests allowed clear phenotypic differentiation of strain 05-Be-005T from the three known Prauserella species. Strain 05-Be-005T represents a novel Prauserella species, for which we propose the name Prauserella muralis sp. nov., with the type strain 05-Be-005T (=CCUG 57426T =NRRL B-24780T =CCM 7635T=DSM 45305T).

The taxonomic positions of three Lechevalieria-like strains isolated from hyperarid soils of the Atacama Desert, Chile, were established by using a polyphasic approach. The organisms had chemical and morphological properties consistent with their classification in the genus Lechevalieria. They formed a distinct subclade in the Lechevalieria 16S rRNA gene clade and were most closely related to the type strain of Lechevalieria xinjiangensis. DNA–DNA relatedness data showed that each of the isolates and Lechevalieria xinjiangensis DSM 45081T belong to distinct genomic species. The new isolates and the type strains of recognized Lechevalieria species were readily distinguished based on a number of phenotypic properties. A combination of the genotypic and phenotypic data showed that the three isolates represent three novel species of the genus Lechevalieria. The names proposed for these taxa are Lechevalieria atacamensis sp. nov. (type strain C61T =CGMCC 4.5536T =NRRL B-24706T), Lechevalieria deserti sp. nov. (type strain C68T =CGMCC 4.5535T =NRRL B-24707T) and Lechevalieria roselyniae sp. nov. (type strain C81T =CGMCC 4.5537T =NRRL B-24708T).

This work deals with the taxonomic study of a bacterium, strain Tp12T, isolated from caterpillars of the pine processionary moth (Thaumetopoea pityocampa Denis & Schiffermüller, 1775; Lepidoptera, Thaumetopoeidae). The isolate was assigned to the genus Brevibacterium on the basis of a polyphasic taxonomic study, including morphological and biochemical characteristics, 16S rRNA gene sequence analysis, fatty acid analysis and DNA G+C content. The highest 16S rRNA gene sequence similarity to this isolate was approximately 96 %, with the type strains of Brevibacterium album and Brevibacterium samyangense. Cellular fatty acids of the isolate are of the branched type, with the major components being anteiso-C15 : 0 and anteiso-C17 : 0. The DNA G+C content was 69.8 mol%. Although the strain was related to B. album and B. samyangense according to 16S rRNA gene sequence analysis, it differed from any known species of Brevibacterium. Based on this evidence, the novel species Brevibacterium pityocampae sp. nov. is proposed, with strain Tp12T (=DSM 21720T =NCCB 100255T) as the type strain.

A novel actinomycete, designated strain GUI 15T, isolated from the root nodules of a Pisum sativum plant was characterized taxonomically by using a polyphasic approach. The 16S rRNA gene sequence of strain GUI 15T showed highest similarity to Micromonospora pattaloongensis TJ2-2T (98.7 %) and Polymorphospora rubra TT 97-42T (98.5 %). Phylogenetic analysis based on the gyrase B gene also supported the close relationship of these three strains, but indicated that strain GUI 15T should be assigned to the genus Micromonospora. Chemotaxonomic results confirmed the position of the isolate in the genus Micromonospora, but revealed differences at the species level. The novel strain could be distinguished from recognized Micromonospora species by using a combination of physiological and biochemical tests. Based on these observations, strain GUI 15T is considered to represent a novel species of the genus Micromonospora, for which the name Micromonospora pisi sp. nov. is proposed. The type strain is GUI 15T (=DSM 45175T=LMG 24546T).

A Gram-positive, non-spore-forming, aerobic bacterium, designated Gsoil 001T, was isolated from the soil of a ginseng field in Pocheon province, South Korea. On the basis of 16S rRNA gene sequence analysis, strain Gsoil 001T was shown to belong to the genus Kribbella. The closest phylogenetic relatives were Kribbella koreensis LM 161T (99.0 % 16S rRNA gene sequence similarity), Kribbella flavida KACC 20248T (98.8 %) and Kribbella alba (98.8%). Lower similarity (98.4–96.9 %) was found with the other recognized species of the genus Kribbella. The major quinone was MK-9(H4), the major fatty acids were anteiso-C15 : 0 and iso-C16 : 0 and the major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylinositol. The cell-wall peptidoglycan contained ll-2,6-diaminopimelic acid. These chemotaxonomic data and the high genomic DNA G+C content (66.3 mol%) supported the affiliation of strain Gsoil 001T to the genus Kribbella. The results of physiological and biochemical tests differentiated strain Gsoil 001T genotypically and phenotypically from the Kribbella species with validly published names. Therefore, this isolate represents a novel species, for which the name Kribbella ginsengisoli sp. nov. is proposed, with the type strain Gsoil 001T (=KCTC 19134T =DSM 17941T).

A novel carbendazim-degrading actinobacterium, designated djl-6-2T, was isolated from the sludge of a carbendazim wastewater treatment facility in Jiangsu province, China. The morphological and chemotaxonomic properties of the isolate were typical of members of the genus Rhodococcus. Strain djl-6-2T formed a coherent cluster with Rhodococcus qingshengii djl-6T, Rhodococcus baikonurensis DSM 44587T, Rhodococcus erythropolis DSM 43066T and Rhodococcus globerulus DSM 43954T in 16S rRNA gene sequence analysis. The results of DNA–DNA hybridization with the above strains (27.7, 19.3, 18.6 and 10.6 % relatedness, respectively), in combination with differences in biochemical and physiological properties, suggest that strain djl-6-2T should be classified within a novel species of the genus Rhodococcus, for which the name Rhodococcus jialingiae sp. nov. is proposed, with djl-6-2T (=DSM 45257T =CCTCC AB 208292T) as the type strain.

A Gram-positive, rod-shaped, non-spore-forming bacterium (Gsoil 346T) was isolated from the soil of a ginseng field in South Korea and was characterized in order to determine its taxonomic position. On the basis of 16S rRNA gene sequences, strain Gsoil 346T was shown to belong to the genus Nocardioides in the family Nocardioidaceae, with the most closely related species being Nocardioides aquiterrae GW-9T (96.6 % 16S rRNA gene sequence similarity); however, the strain clustered in a distinct branch of the phylogenetic tree with Nocardioides kongjuensis A2-4T (96.2 %), Nocardioides aromaticivorans H-1T (96.1 %), Nocardioides nitrophenolicus NSP41T (96.1 %) and Nocardioides simplex ATCC 15799T (95.9 %). Strain Gsoil 346T was characterized chemotaxonomically and found to have ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan, phosphatidylinositol and phosphatidylglycerol as the major polar lipids, MK-8(H4) as the predominant menaquinone and iso-C16 : 0, C18 : 1 ω9c and C17 : 1 ω8c as the major fatty acids. The G+C content of the genomic DNA of the novel strain was 73.0 mol%. These chemotaxonomic properties supported the placement of strain Gsoil 346T in the genus Nocardioides. The results of physiological and biochemical tests, along with the phylogenetic analysis, allowed strain Gsoil 346T to be differentiated genotypically and phenotypically from recognized species of the genus Nocardioides. Therefore, strain Gsoil 346T represents a novel species, for which the name Nocardioides panacisoli sp. nov. is proposed, with Gsoil 346T (=KCTC 19470T=DSM 21348T) as the type strain.

A Gram-stain-positive, coccoid, non-endospore-forming actinobacterium (Sj14aT) was isolated from the air of a duck barn on tryptone soy agar after filter sampling. On the basis of 16S rRNA gene sequence similarity studies, strain Sj14aT was shown to belong to the genus Dietzia and was most closely related to Dietzia schimae (98.7 %), Dietzia cercidiphylli (98.4 %) and Dietzia maris (98.6 %). The major menaquinone was MK-8 (H2). The polar lipid profile included the major components diphosphatidylglycerol, phosphatidylglycerol, an unknown polar lipid and two unknown glycolipids. Phosphatidylinositol and two phosphatidylinositol mannosides, as well as several other unknown lipids, were also detected. The polyamine pattern contained the major compounds spermine and spermidine. The fatty acid profile, containing C16 : 0, C17 : 0, C18 : 1 ω9c and 10-methyl C18 : 0 as major fatty acids, supported the affiliation of strain Sj14aT to the genus Dietzia. The results of physiological and biochemical tests and DNA–DNA hybridizations allowed a clear phenotypic differentiation of strain Sj14aT from the most closely related species of the genus Dietzia. Strain Sj14aT represents a novel species, for which the name Dietzia aerolata sp. nov. is proposed, with the type strain Sj14aT (=DSM 45334T =CCM 7659T).

A novel actinomycete strain, ABOT, isolated from copper-polluted sediments showed remarkable copper resistance as well as high bioaccumulation abilities. Classical taxonomic methods, including chemotaxonomy and molecular techniques, were used to characterize the isolate. Strain ABOT developed a honey-yellow substrate mycelium on all ISP media tested. Abundant, white, aerial mycelium was only formed on ISP 2, 5 and 7 and MM agar. Both types of hyphae fragmented into squarish rod-shaped elements. The aerial mycelium displayed spore-like structures with smooth surfaces in long, straight to flexuous chains. The organism has a type-IV cell wall lacking mycolic acids and type-A whole-cell sugar pattern (meso-diaminopimelic acid, arabinose and galactose) in addition to a phospholipid type-II profile. 16S rRNA gene sequence studies indicated that this organism is a member of the family Pseudonocardiaceae and that it forms a monophyletic clade with Amycolatopsis eurytherma NT202T. The DNA–DNA relatedness of strain ABOT to A. eurytherma DSM 44348T was 39.5 %. It is evident from these genotypic and phenotypic data that strain ABOT represents a novel species in the genus Amycolatopsis, for which the name proposed is Amycolatopsis tucumanensis sp. nov. The type strain is ABOT (=DSM 45259T =LMG 24814T).

A previously unidentified, slowly growing, scotochromogenic Mycobacterium species, represented by strain 31118T, was discovered during hsp65 sequence-based reidentification of Korean clinical isolates that had been previously identified as Mycobacterium scrofulaceum by conventional biochemical tests. Although the 16S rRNA gene sequence of strain 31118T was identical to that of the recently described Mycobacterium seoulense, phylogenetic analyses based on three independent alternative targets (rpoB, hsp65 and the 16S–23S internal transcribed spacer) showed that it was closely related to M. seoulense but was a distinct phylogenetic entity. Furthermore, the phenetic characteristics of this strain were more similar to those of M. scrofulaceum than to those of M. seoulense. Taken together, these results support the conclusion that this strain represents a novel mycobacterium species, for which the name Mycobacterium paraseoulense sp. nov. is proposed. The type strain is 31118T (=DSM 45000T =KCTC 19145T).

An actinomycete strain, ID05-A0528T, was isolated using the SDS-yeast extract pre-treatment method from soil under mahogany (Swietenia mahogani) trees in West Timor, Indonesia, and was examined by using a polyphasic taxonomic approach. Chemotaxonomic and phylogenetic characterizations demonstrated that the novel strain belongs to the genus Dietzia. 16S rRNA gene sequencing studies showed that the strain was related to Dietzia cinnamea (97.2 %). Results of phenotypic and phylogenetic analyses determined that strain ID05-A0528T is different from the known species of the genus Dietzia. It is proposed that the isolate should be classified as a representative of a novel species of the genus Dietzia, with the name Dietzia timorensis sp. nov. The type strain is ID05-A0528T (=BTCC B-560T =NBRC 104184T).

A methanogenic archaeon, strain MK4T, was isolated from ancient permafrost after long-term selective anaerobic cultivation. The cells were rods, 2.0–8.0 μm long and 0.40–0.45 μm wide, and stained Gram-negative. Optimal growth was observed at 28 °C and pH 7.0–7.2 and in 0.05 M NaCl. The isolate used H2 plus CO2, methylamine plus H2 and methanol plus H2 as sources for growth and methanogenesis. Phylogenetic analysis of the 16S rRNA gene sequence of the strain showed close affinity with Methanobacterium bryantii (similarity >99 % to the type strain). On the basis of the level of DNA–DNA hybridization (62 %) between strain MK4T and Methanobacterium bryantii VKM B-1629T and phenotypic and phylogenetic differences, strain MK4T was assigned to a novel species of the genus Methanobacterium, Methanobacterium veterum sp. nov., with the type strain MK4T (=DSM 19849T =VKM B-2440T).

A rod-shaped Gram-staining-negative, non-motile, aerobic and fucoidan-digesting strain, designated TC2T, was isolated from marine algae collected from the coast of the Sea of Okhotsk at Abashiri, Hokkaido, Japan. The bacterium formed yellow, translucent, circular and convex colonies. Comparative 16S rRNA gene sequence analysis indicated that the strain belonged to the genus Flavobacterium, with the highest sequence similarities of 97.1 to 97.3 % to the type strains of Flavobacterium frigidarium, Flavobacterium frigoris, Flavobacterium limicola and Flavobacterium psychrolimnae. DNA–DNA relatedness values between strain TC2T and the above-mentioned species were lower than 28 %. The genomic DNA G+C content was 33.9 mol%. The major respiratory quinone was menaquinone-6 and the predominant fatty acids were iso-C15 : 1 G, iso-C15 : 0, iso-C15 : 0 3-OH and summed feature 3 (which comprises iso-C15 : 0 2-OH and/or C16 : 1 ω7c). Strain TC2T could be differentiated from related species by several phenotypic characteristics. Thus, on the basis of these results, strain TC2T represents a novel species of the genus Flavobacterium, for which the name Flavobacterium algicola sp. nov. is proposed. The type strain is TC2T (=NBRC 102673T =CIP 109574T).

An aerobic, yellow-pigmented, Gram-staining-negative bacterium, designated strain H7T, was isolated from sediment of Gazwa stream in Jinju city, South Korea. Cells of strain H7T were non-motile, straight rods that produced flexirubin pigments and showed catalase- and oxidase activities. Strain H7T contained iso-C15 : 0, anteiso-C15 : 0 and summed feature 3 (comprising C16 : 1 ω7c and/or iso-C15 : 0 2-OH) as the main fatty acids and menaquinone-6 (MK-6) as the major isoprenoid quinone. The DNA G+C content of strain H7T was 37.2 mol%. Comparative 16S rRNA gene sequence analysis showed that strain H7T formed a distinct phyletic line within the genus Flavobacterium. Based on 16S rRNA gene sequence similarities, the novel strain was most closely related to Flavobacterium limicola ST-82T and Flavobacterium resistens BD-b365T with 97.2 % and 97.0 % gene sequence similarities, respectively. The DNA–DNA relatedness of the novel strain with these species was only 18 % and 14 %, respectively. On the basis of phenotypic data and molecular properties, strain H7T represents a novel species within the genus Flavobacterium, for which the name Flavobacterium fluvii sp. nov. is proposed. The type strain is H7T (=KACC 12818T=DSM 19978T).

A novel marine, Gram-staining-negative, yellow-pigmented, rod-shaped bacterial strain, designated CNU004T, was isolated from a seawater sample collected on the coastline of Jeju Island, South Korea. The strain was strictly aerobic, non-flagellated, non-gliding and oxidase- and catalase-positive. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CNU004T belongs to a distinct lineage in the family Flavobacteriaceae. Strain CNU004T exhibited levels of 16S rRNA gene sequence similarity of 93.8–93.9 % to its nearest phylogenetic neighbours, members of the genera Gaetbulibacter, Yeosuana and Algibacter. The new isolate required sea salts or artificial seawater for growth. The optimum ranges of temperature and pH for growth were 30–35 °C and pH 7.0–8.0. The DNA G+C content of strain CNU004T was 37.7 mol%. The major fatty acids were iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH and iso-C15 : 0 3-OH. Menaquinone-6 was the major respiratory quinone. Zeaxanthin was the major carotenoid pigment produced, and flexirubin-type pigments were not produced. Strain CNU004T was able to degrade starch and agar. Based on its phenotypic and genotypic characteristics and on the phylogenetic evidence presented, strain CNU004T is considered to represent a novel species of a new genus in the family Flavobacteriaceae, for which the name Hyunsoonleella jejuensis gen. nov., sp. nov. is proposed. The type strain of Hyunsoonleella jejuensis sp. nov. is CNU004T (=KCTC 22242T =DSM 21035T).

Two Gram-staining-negative bacterial strains, designated 3A10T and ECP37T, were isolated from sediment samples collected from an industrially contaminated site in northern Portugal. These two organisms were rod-shaped, non-motile, aerobic, catalase- and oxidase-positive and formed yellow colonies. The predominant fatty acids were iso-C15 : 0, anteiso-C15 : 0, iso-C17 : 1 ω9c and iso-C17 : 0 3-OH. The G+C content of the DNA of strains 3A10T and ECP37T was 43 and 34 mol%, respectively. The major isoprenoid quinone of the two strains was MK-6. 16S rRNA gene sequence analysis revealed that strains 3A10T and ECP37T were members of the family Flavobacteriaceae and were related phylogenetically to the genus Chryseobacterium. Strain 3A10T showed 16S rRNA gene sequence similarity values of 97.2 and 96.6 % to the type strains of Chryseobacterium antarcticum and Chryseobacterium jeonii, respectively; strain ECP37T showed 97.3 % similarity to the type strain of Chryseobacterium marinum. DNA–DNA hybridization experiments revealed levels of genomic relatedness of <70 % between strains 3A10T and ECP37T and between these two strains and the type strains of C. marinum, C. antarcticum and C. jeonii, justifying their classification as representing two novel species of the genus Chryseobacterium. The names proposed for these organisms are Chryseobacterium palustre sp. nov. (type strain 3A10T =LMG 24685T =NBRC 104928T) and Chryseobacterium humi sp. nov. (type strain ECP37T =LMG 24684T =NBRC 104927T).

A Gram-stain-negative, heterotrophic, aerobic, non-spore-forming and non-motile bacterial strain, designated LM5T, was isolated from activated sludge from a sequencing batch reactor for the treatment of effluents contaminated by malachite green. The taxonomy of strain LM5T was studied by phenotypic and phylogenetic methods. Strain LM5T formed orange colonies on R2A and YP plates. Cells were rods, 0.4–0.6 μm in diameter and 0.8–1.2 μm in length. Growth occurred at 10–35 °C (optimum, 20–25 °C), at pH 5.5–9.5 (optimum, pH 6.5–7.5) and in the presence of 0–2 % (w/v) NaCl (optimum, 0.5 %). Oxidase and catalase activities were present. Flexirubin-type pigments were present, but extracellular glycans were absent. MK-6 was the major respiratory quinone. The major fatty acids were iso-C15 : 0 (28.3 %) and iso-C17 : 1 ω9c (13.8 %). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain LM5T was a member of the genus Flavobacterium with highest sequence similarity to Flavobacterium soli DS-6T (93.2 %) and Flavobacterium lindanitolerans IP-10T (92.9 %). Together with F. lindanitolerans IP-10T, strain LM5T formed a distinct lineage in the phylogenetic tree. The DNA G+C content was 52±0.6 mol% (HPLC), which is significantly higher than that of other species of the genus Flavobacterium (30–41 mol%). Based on phylogenetic and phenotypic evidence, strain LM5T is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium caeni sp. nov. is proposed; the type strain is LM5T (=CGMCC 1.7031T=NBRC 104239T).

A novel xylan-degrading bacterium, designated XDB9T, was isolated from forest humus collected from Gyeryong Mountain in Korea. Cells were Gram-positive, aerobic, motile and endospore-forming rods. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain XDB9T was most closely related to members of the genus Lysinibacillus. 16S rRNA gene sequence similarities between strain XDB9T and the type strains of species of the genus Lysinibacillus ranged from 98.0 to 98.5 %. The cell-wall peptidoglycan type of strain XDB9T was A4α, which is based on l-Lys–d-Asp. Strain XDB9T contained iso-C15 : 0 and C16 : 1 ω7c alcohol as the major fatty acids and MK-7 as the predominant menaquinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content was 37.2 mol%. The DNA–DNA hybridization results and differential phenotypic properties showed that strain XDB9T could be distinguished from recognized species of the genus Lysinibacillus. It was concluded that strain XDB9T represents a new taxon for which the name Lysinibacillus xylanilyticus sp. nov. is proposed. The type strain is XDB9T (=KCTC 13423T=CCUG 57438T).