- Volume 60, Issue 9, 2010

A bacterial strain, designated PW21T, was isolated from root nodules of Prosopis farcta in Tunisia. Phylogenetic analysis based on 16S rRNA gene sequences placed the isolate into the genus Paenibacillus, with its closest relatives being Paenibacillus glycanilyticus DS-1T and Paenibacillus castaneae Ch-32T with identity values of 96.9 %. DNA–DNA hybridization measurements showed values of less than 25 % with respect to these two species. The isolate was a Gram-variable, motile and sporulating rod. Catalase activity was positive and oxidase activity was weakly positive. Aesculin, CM-cellulose, xylan and starch were hydrolysed but casein and gelatin were not. Acetoin production was weakly positive and nitrate reduction was negative. Urease production was negative. Growth was supported by many carbohydrates and organic acids as carbon sources. MK-7 was the predominant menaquinone and anteiso-C15 : 0, iso-C16 : 0 and iso-C15 : 0 were the major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, a glycolipid, six phospholipids, an unidentified lipid and two unknown aminophosphoglycolipids. meso-Diaminopimelic acid was not detected in the peptidoglycan. The DNA G+C content of the isolate was 52.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain PW21T should be considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus prosopidis sp. nov. is proposed. The type strain is PW21T (=LMG 25259T =CECT 7506T =DSM 22405T).

Four Gram-positive, catalase-negative, short rod-shaped or coccoid, heterofermentative lactic acid bacterial strains (2L24P13T, 1L48P15, 1L24P31 and 1L24P34) with unusual phenotypic and genotypic properties were isolated from submerged fermenting cassava on MRS agar. All strains were motile, grew at 15 °C, produced dl-lactic acid from glucose with gas formation and produced ammonia from arginine. Acid was produced from d-fructose, d-galactose, d-glucose, lactose, maltose, d-mannose, melibiose, d-raffinose, sucrose, N-acetylglucosamine and d-mannitol, but not from d-arabinose or xylose. 16S rRNA gene sequence analysis revealed that the strains belonged to the genus Weissella and were most closely related to Weissella ghanensis LMG 24286T . Low DNA−DNA reassociation values were obtained between the isolates and W. ghanensis DSM 19935T. Based on the genetic and phenotypic results, the strains are considered to represent a novel species, for which the name Weissella beninensis sp. nov. is proposed. The type strain is 2L24P13T (=DSM 22752T=LMG 25373T).

Two Gram-stain-positive strains, WPCB074T and WPCB165, were isolated from fresh water collected from the Woopo wetland (Republic of Korea). Both strains were strictly aerobic, motile, endospore-forming rods. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains WPCB074T and WPCB165 belonged to the genus Bacillus and that strain WPCB074T was most closely related to Bacillus solisalsi YC1T (98.4 % sequence similarity), B. barbaricus V2-BIII-A2T (97.7 %), B. macauensis ZFHKF-1T (96.9 %), B. arsenicus Con a/3T (96.4 %) and B. gelatini LMG 21880T (95.1 %). The 16S rRNA gene sequences of strains WPCB074T and WPCB165 differed at one position (99.9 % similarity), suggesting that these two strains constitute a single species. DNA–DNA relatedness between strain WPCB074T and the type strains of B. solisalsi, B. barbaricus, B. macauensis, B. arsenicus and B. gelatini were 26, 17, 20, 14 and 7 %, respectively. Strain WPCB074T was characterized by having cell-wall peptidoglycan based on meso-diaminopimelic acid, MK-7 as the predominant menaquinone and iso-C15 : 0 and anteiso-C15 : 0 as the major fatty acids. The DNA G+C content of strain WPCB074T was 41.9 mol%. On the basis of phenotypic properties, phylogeny and genomic distinctiveness, strain WPCB074T represents a novel species of the genus Bacillus for which the name Bacillus rigui sp. nov. is proposed. The type strain is WPCB074T (=KCTC 13278T =JCM 16348T).

A Listeria-like strain isolated in Austria from pre-cut lettuce fitted the description of the genus Listeria although it could not be assigned to any of the known species. Comparison of the rrs gene (encoding 16S rRNA) sequence and gene content by DNA-array indicated affiliation to the genus Listeria. Phylogenetic distance from known species of the genus Listeria indicated that it represents a novel species. Since it can be differentiated from all other known species of the genus Listeria by using phenotypic tests, the name Listeria rocourtiae sp. nov. is proposed for the novel species. The type strain is CIP 109804T (=DSM 22097T =Allerberger 700284/02T). The type strain is avirulent as assessed by cell culture assays and inoculation of mice.

Two strictly anaerobic, proteolytic bacterial strains, designated strain D3RC-2T and D3RC-3r, were isolated from a cellulose-degrading mixed culture enriched from yak rumen content. The strains were Gram-stain negative and non-spore-forming with cell sizes of 0.5–0.8×0.6–2.0 μm. The temperature range for growth was 24–46 °C (optimum 38–39 °C) and the pH range was between 5.6 and 8.7 (optimum 7.0–7.3). Both strains used soya peptone, tryptone, l-phenylalanine, l-leucine, l-methionine, l-serine, l-valine, l-threonine and l-histidine as carbon and nitrogen sources, but did not use any of the saccharides tested. The major fermentation products from PY medium were acetate, propionate and iso-butyrate. The DNA G+C contents of strains D3RC-2T and D3RC-3r were 41.0±0.1 mol% and 41.3±0.1 mol% (HPLC), respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the two strains represented a new phyletic sublineage within the family Clostridiaceae, with <93.8 % 16S rRNA gene sequence similarity to recognized species. On the basis of the phenotypic, genotypic and physiological evidence, strains D3RC-2T and D3RC-3r are proposed as representing a novel species of a new genus, for which the name Proteiniclasticum ruminis gen. nov., sp. nov. is proposed. The type strain of the type species is D3RC-2T (=AS 1.5057T=JCM 14817T).

A thermophilic strain, designated YIM 10002T, was isolated from a soil sample of Big Empty Volcano in Tengchong county, Yunnan province, south-west China, and a polyphasic approach was used to investigate its taxonomic position. Strain YIM 10002T formed endospores on both aerial and substrate mycelia. Whole-cell hydrolysates contained meso-diaminopimelic acid, ribose, xylose and glucose. The major fatty acids were iso-C15 : 0 and anteiso-C15 : 0. The predominant menaquinone was MK-9. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides, together with some unknown phospholipids. The G+C content of its genomic DNA was 48.6 mol%. All of these chemotaxonomic data together with morphological characters consistently assigned strain YIM 10002T to the genus Laceyella. 16S rRNA gene sequence analysis showed that strain YIM 10002T was most closely related to Laceyella sacchari KCTC 9790T and Laceyella putida KCTC 3666T (99.9 and 98.0 % 16S rRNA gene sequence similarity, respectively). However, strain YIM 10002T showed relatively low DNA–DNA relatedness (34.0 and 39.0 %, respectively) with the above strains. Therefore, strain YIM 10002T represents a novel species of the genus Laceyella, for which the name Laceyella tengchongensis sp. nov. is proposed. The type strain is YIM 10002T (=DSM 45262T =CCTCC AA 208050T).

An obligately anaerobic, Gram-positive, spore-forming bacterial strain, designated SL206T, was isolated from pear orchard soils. Strain SL206T cells were straight or slightly curved rods, with motility by peritrichate flagella. Cell walls contained meso-diaminopimelic acid; wall sugars were glucose, rhamnose and mannose. The major fatty acids were C16 : 0, C18 : 1 ω9c and summed feature 10 (containing C18 : 1 ω11c/9t/6t). API 20A reactions were negative for oxidase, catalase and acid production from l-rhamnose, sucrose, trehalose, d-xylose, melezitose, salicin and d-sorbitol, and positive for acid production from d-glucose, sucrose, maltose, d-mannose and raffinose. Glucose was fermented to acetate, butyrate, CO2, H2 and ethanol in culture. The G+C content of the genomic DNA was 31.1 mol%. Based on comparative 16S rRNA gene sequence analysis, the isolate belonged to the genus Clostridium and formed a clade with Clostridium pasteurianum. The species most closely related to strain SL206T were C. pasteurianum (98.6 % similarity) and Clostridium acidisoli (97.8 % similarity). In DNA–DNA relatedness studies, the isolate had 59.5 % relatedness with C. pasteurianum and thus represented a unique species. On the basis of these studies, strain SL206T (=KCTC 5449T =JCM 14858T) is proposed to represent the type strain of a novel species, Clostridium arbusti sp. nov.

The taxonomic positions of two gamma- and UV-ray-resistant strains isolated from radiation-polluted soil in north-west China were determined in a polyphasic study. The organisms, designated R12T and R13T, were Gram-stain-positive, non-spore-forming cocci, which contained MK-8 as the major respiratory quinone and C16 : 1 ω7c and C16 : 0 as major fatty acids. The cell walls of strains R12T and R13T contained ornithine. Phylogenetic analysis based on 16S rRNA gene sequences and DNA–DNA hybridizations showed that strains R12T and R13T are members of novel species belonging to the genus Deinococcus, with Deinococcus radiodurans DSM 20539T as the closest relative. The isolates R12T and R13T shared 97 and 97.1 % 16S rRNA gene similarity, respectively, and 29.5 and 33.3 % DNA–DNA relatedness, respectively, with D. radiodurans DSM 20539T. The DNA G+C contents of isolates R12T and R13T were 66.7 and 63.8 %, respectively. On the basis of phenotypic tests and other results, two species, Deinococcus wulumuqiensis sp. nov. (type strain R12T =CGMCC 1.8884T =NBRC 105665T) and Deinococcus xibeiensis sp. nov. (type strain R13T =CGMCC 1.8885T =NBRC 105666T), are proposed.

Twenty coagulase-negative Staphylococcus strains displaying α-haemolysis (δ-haemolysin) on sheep-blood agar were isolated from the noses of different pigs in Switzerland. The strains were Gram-stain-positive, non-motile cocci, catalase-positive and coagulase-negative. Sequence analysis of the 16S rRNA gene, sodA, rpoB, dnaJ and hsp60 and phylogenetic characteristics revealed that the strains showed the closest relatedness to Staphylococcus microti CCM 4903T and Staphylococcus muscae DSM 7068T. The strains can be differentiated from S. microti by the absence of mannose fermentation and arginine arylamidase and from S. muscae by the absence of β-glucuronidase activity and production of alkaline phosphatase. The chosen type strain ARI 262T shared 20.1 and 31.9 % DNA relatedness with S. microti DSM 22147T and S. muscae CCM 4903T, respectively, by DNA–DNA hybridization. iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0 were the most common fatty acids. Cell-wall structure analysis revealed the peptidoglycan type A3α l-Lys–Gly2–l-Ser–Gly (type A11.3). The presence of teichoic acid was determined by sequencing the N-acetyl-β-d-mannosaminyltransferase gene tarA, which is involved in biosynthesis of ribitol teichoic acid. Menaquinone 7 (MK-7) was the predominant respiratory quinone. The G+C content of ARI 262T was 38.8 mol%. The isolated strains represent a novel species of the genus Staphylococcus, for which we propose the name Staphylococcus rostri sp. nov. The type strain is ARI 262T (=DSM 21968T =CCUG 57266T) and strain ARI 602 (=DSM 21969 =CCUG 57267) is a reference strain.

A novel thermophilic, strictly anaerobic, chemo-organotrophic bacterium, designated MCT, was isolated from a geothermally heated sediment of a marine hydrothermal system at Palaeochory Bay, Milos, Greece. Cells of strain MCT were rods of variable length (4–12 μm) and width (0.2–0.3 μm), occurring as single cells or forming large aggregates that were visible as flocs. Strain MCT grew optimally at pH 7.0 and 60 °C and with 3 % (w/v) NaCl. Strain MCT grew chemo-organoheterotrophically and fermented peptides and di- and polysaccharides in the presence of 0.1 g yeast extract l−1. The DNA G+C content of strain MCT was 43.3 mol%. Phylogenetic analysis of the 16S rRNA gene sequence placed strain MCT within the genus Caldithrix. However, strain MCT possessed certain phenotypic features that differentiated it from the type strain of the only species of the genus Caldithrix described to date. On the basis of phylogenetic and phenotypic characteristics, it is proposed that strain MCT represents a novel species, Caldithrix palaeochoryensis sp. nov. The type strain is MCT (=DSM 21940T =VKM B-2536T). In addition, an emended description of the genus Caldithrix is presented.

The protein profiles, genomic amplified fragment length polymorphism patterns and 16S rRNA and cpn60 gene sequences of a diverse collection of 26 Bacteroides ureolyticus strains, along with published data on their DNA base, respiratory quinone and cellular fatty acid compositions, were used to reassess the taxonomy of this bacterial species. The results demonstrate that this organism is most appropriately allocated in the genus Campylobacter. The presence of much higher amounts of 18 : 1ω7c in its cellular fatty acid profile and its ability to digest gelatin and casein are the characteristics that differentiate it from present species of the genus Campylobacter. Therefore we propose to reclassify this species incertae sedis into the genus Campylobacter as Campylobacter ureolyticus with strain LMG 6451T (=CCUG 7319T =NCTC 10941T) as the type strain.

A Gram-negative, rod-shaped, non-pigmented, non-spore-forming bacterial strain that was motile by a single polar flagellum, designated A1-9T, was isolated from Daqing reservoir in north-east China and its taxonomic position was studied using a polyphasic approach. Strain A1-9T was non-halophilic, strictly aerobic and heterotrophic and lacked carotenoids, internal membranes and genes for photosynthesis (puf genes). Strain A1-9T grew at 10–40 °C (optimum, 25–30 °C) and pH 5.5–9.0 (optimum, pH 6.0–6.5) and tolerated up to 1.0 % NaCl (w/v). Neither phototrophic nor fermentative growth was observed. The predominant ubiquinone was Q-10 and the major fatty acid was C18 : 1 ω7c (70 %). The DNA G+C content was 64.4 mol% (T m). Phylogenetic analyses based on the 16S rRNA gene sequence revealed that strain A1-9T, together with Catellibacterium nectariphilum AST4T, formed a deep line within the ‘Rhodobacter clade’ of the family Rhodobacteraceae and strain A1-9T showed 94.2 % 16S rRNA gene sequence similarity to C. nectariphilum AST4T. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain A1-9T is considered to represent a novel species of the genus Catellibacterium, for which the name Catellibacterium aquatile sp. nov. is proposed. The type strain is A1-9T (=CGMCC 1.7029T =NBRC 104254T).

A Gram-negative, beige-pigmented, aerobic, motile, club-shaped bacterium, designated strain CC-SBABM117T, was isolated from the stipe of the edible mushroom Agaricus blazei Murrill. 16S rRNA gene sequence analysis demonstrated that the strain shared <93 % similarity with the type strains of species in the genera Pannonibacter, Methylopila, Nesiotobacter and Stappia. The organism was unable to produce acid from carbohydrates, but utilized a number of organic acids and amino acids. Ubiquinone 10 (Q-10) was the major respiratory quinone and C18 : 1 ω7c, C19 : 0 cyclo ω8c, C16 : 0 and C18 : 0 were the predominant fatty acids. The predominant polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content of strain CC-SBABM117T was 62.7 mol%. On the basis of 16S rRNA gene sequence analysis and chemotaxonomic and physiological data, strain CC-SBABM117T is considered to represent a novel species of a new genus, for which the name Agaricicola taiwanensis gen. nov., sp. nov. is proposed. The type strain of Agaricicola taiwanensis is CC-SBABM117T (=BCRC 17964T =CCM 7684T).

Two strains of aluminium-tolerant bacteria, SA33T and 7A078, were isolated from Chinese water chestnut (Eleocharis dulcis) growing in highly acidic swamps (pH 2–4) in actual acid sulfate soil areas of Vietnam (SA33T) and Thailand (7A078). The strains were Gram-negative, aerobic, non-spore-forming rods, 0.6–0.7 μm wide and 1.3–1.7 μm long. These strains showed good growth at pH 3.0–8.0 and 17–37 °C. The organisms contained ubiquinone Q-8 as the predominant isoprenoid quinone and C16 : 0, C18 : 1 ω7c and C17 : 0 cyclo as the major fatty acids. Their fatty acid profiles were similar to those reported for other Burkholderia species. The DNA G+C content of these strains was 64 mol%. On the basis of 16S rRNA gene sequence similarity, the strains were shown to belong to the genus Burkholderia. Although the 16S rRNA gene sequence similarity values calculated for strain SA33T to 7A078 and the type strains of Burkholderia kururiensis, B. sacchari and B. tuberum were 100, 97.3, 97.1 and 97.0 %, respectively, strains SA33T and 7A078 formed a group that was distinct in the phylogenetic trees; the DNA–DNA relatedness of strain SA33T to 7A078 and these three type strains were respectively 90, 47, 46 and 45 %. The results of physiological and biochemical tests, including whole-cell protein pattern analysis, allowed phenotypic differentiation of these strains from described Burkholderia species. Therefore, strains SA33T and 7A078 represent a novel species, for which the name Burkholderia acidipaludis sp. nov. is proposed. The type strain is SA33T (=NBRC 101816T =VTCC-D6-6T). Strain 7A078 (=NBRC 103872 =BCC 36999) is a reference strain.

Two clinical Aeromonas strains (A2-50T and A2-67T) recovered from the wounds of two patients in Taiwan could not be assigned to any known species of this genus based on their 16S rRNA gene sequences, which showed similarities of 99.6–99.8 % to those of the type strains of Aeromonas caviae, A. trota and A. aquariorum. The rpoD phylogenetic tree allocated these strains to two novel and independent phylogenetic lines, the neighbouring species being A. caviae, the type strain of which showed 93.2 % similarity (56 bp differences) to strain A2-50T and 92.2 % (63 bp differences) to strain A2-67T. A multilocus phylogenetic analysis of five housekeeping genes (gyrB, rpoD, recA, dnaJ and gyrA; 3684 bp) confirmed that the two strains formed independent phylogenetic lineages within the genus. These data, together with phenotypic characterization and DNA–DNA reassociation results, revealed that these strains represent novel Aeromonas species, for which the names Aeromonas taiwanensis sp. nov. (type strain A2-50T =CECT 7403T =LMG 24683T) and Aeromonas sanarellii sp. nov. (type strain A2-67T =CECT 7402T =LMG 24682T) are proposed.

A Gram-negative, non-spore-forming, motile, facultatively anaerobic bacterium, designated strain MJ06T, was isolated from oil-contaminated soil and was characterized taxonomically by using a polyphasic approach. Strain MJ06T contained ubiquinone Q-8 as the major respiratory lipoquinone, putrescine as the predominant polyamine and phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. The G+C content of the genomic DNA of strain MJ06T was 66.2 mol%. The major fatty acids were summed feature 4 (C16 : 1 ω7c and/or iso-C15 : 0 2-OH; 32.5 %), C16 : 0 (22.8 %) and summed feature 7 (one or more of C18 : 1 ω7c, C18 : 1 ω9t and C18 : 1 ω12t; 14.9 %). Comparative 16S rRNA gene sequence analysis showed that strain MJ06T belonged to the family Alcaligenaceae, class Betaproteobacteria, and joined the evolutionary radiation enclosed by the genus Castellaniella. Levels of 16S rRNA gene sequence similarity between strain MJ06T and its phylogenetically closest relatives, Castellaniella denitrificans NKNTAUT, Castellaniella defragrans 54PinT, Castellaniella ginsengisoli DCY36T and Castellaniella caeni Ho-11T, were 98.6, 98.3, 97.8 and 97.3 %, respectively. Levels of similarity between strain MJ06T and the type strains of all other recognized species in the family Alcaligenaceae were below 95.0 %. Strain MJ06T exhibited relatively low levels of DNA–DNA relatedness with respect to C. defragrans DSM 12141T (52 %), C. denitrificans DSM 11046T (31 %), C. ginsengisoli KCTC 22398T (18 %) and C. caeni KCTC 12197T (15 %). On the basis of its phenotypic and genotypic properties together with phylogenetic distinctiveness, strain MJ06T is considered to represent a novel species of the genus Castellaniella, for which the name Castellaniella daejeonensis sp. nov. is proposed. The type strain is MJ06T (=KCTC 22454T =JCM 16240T).

A non-motile, rod-shaped and aerobic marine bacterium, designated strain KYW314T, was isolated from seawater collected from the South Sea, Republic of Korea. Cells were Gram-negative, catalase- and oxidase-positive and had proteolytic activity. The major fatty acids were iso-C15 : 0 (17.2 %), C16 : 0 (11.1 %), iso-C17 : 0 (10.5 %) and C18 : 1 ω7c (10.0 %). The DNA G+C content was 56.4 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain KYW314T formed a lineage within the genus Pseudidiomarina (95.4–97.3 % sequence similarity) and a distinct branch within the clade containing Pseudidiomarina taiwanensis PIT1T and Pseudidiomarina sediminum c121T. Phenotypic characteristics could distinguish strain KYW314T from members of the genus Pseudidiomarina. On the basis of the data presented, strain KYW314T represents a novel species, for which the name Pseudidiomarina aestuarii sp. nov. is proposed. The type strain is KYW314T (=KCTC 22740T =JCM 16344T).

A novel Gram-negative, rod-shaped, motile, non-spore-forming bacterial strain, CMST, isolated from soil was characterized using phenotypic and molecular taxonomic methods. 16S rRNA gene sequence analysis revealed that the organism belongs phylogenetically to the genus Pseudomonas. Pseudomonas monteilii, P. plecoglossicida and P. mosselii were the most closely related species, with 16S rRNA gene sequence similarities to the respective type strains of 99.79, 99.73 and 99.59 %. Relatively low gyrB gene sequence similarities (<90 %) and DNA–DNA reassociation values (<51 %) were obtained between the strain and its phylogenetically closest neighbours. The G+C content of strain CMST was 62.7 mol%. The major cellular fatty acids were C18 : 1 ω7c, summed feature 3 (C16 : 1 ω7c and/or iso-C15 : 0 2-OH), C16 : 0 and C10 : 0 3-OH. Based on the phenotypic and genetic evidence, the strain is suggested to represent a novel species, for which the name Pseudomonas taiwanensis sp. nov. is proposed. The type strain is CMST (=BCRC 17751T =DSM 21245T).

An oval to rod-shaped, phototrophic, purple non-sulfur bacterium, strain JA192T, was isolated from an enrichment culture of a pasteurized rhizosphere soil sample from a field cultivated with jowar (sorghum) collected from Godumakunta village near Hyderabad, India. Strain JA192T is Gram-negative, motile and produces endospores. Phylogenetic analysis on the basis of 16S rRNA gene sequences showed that the strain JA192T is closely related to Rhodobacter sphaeroides 2.4.1T (99.9 % sequence similarity), Rba. megalophilus JA194T (99.8 %) and Rba. azotoformans KA25T (98.1 %) and clusters with other species of the genus Rhodobacter of the family Rhodobacteraceae. However, DNA–DNA hybridization with Rba. sphaeroides DSM 158T, Rba. megalophilus JA194T and Rba. azotoformans JCM 9340T showed relatedness of only 38–57 % with respect to strain JA192T. On the basis of 16S rRNA gene sequence analysis, DNA–DNA hybridization data and morphological, physiological and chemotaxonomic characters, strain JA192T represents a novel species of the genus Rhodobacter, for which the name Rhodobacter johrii sp. nov. is proposed. The type strain is JA192T (=DSM 18678T =JCM 14543T =MTCC 8172T).