- Volume 65, Issue Pt_6, 2015

A novel, strictly anaerobic, hydrogenotrophic methanogen, strain E09F.3T, was isolated from a commercial biogas plant in Germany. Cells of E09F.3T were Gram-stain-negative, non-motile, slightly curved rods, long chains of which formed large aggregates consisting of intertwined bundles of chains. Cells utilized H2+CO2 and, to a lesser extent, formate as substrates for growth and methanogenesis. The optimal growth temperature was around 40 °C; maximum growth rate was obtained at pH around 7.0 with approximately 6.8 mM NaCl. The DNA G+C content of strain E09F.3T was 39.1 mol%. Phylogenetic analyses based on 16S rRNA and mcrA gene sequences placed strain E09F.3T within the genus Methanobacterium. On the basis of 16S rRNA gene sequence similarity, strain E09F.3T was closely related to Methanobacterium congolense CT but morphological, physiological and genomic characteristics indicated that strain E09F.3T represents a novel species. The name Methanobacterium aggregans sp. nov. is proposed for this novel species, with strain E09F.3T ( = DSM 29428T = JCM 30569T) as the type strain.

A novel endophytic actinobacterium, designated strain YIM 64602T, was isolated from healthy stems of Tripterygium wilfordii. It grew at 15–40 °C, pH 6.0–9.0 and in the presence of 0–3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain YIM 64602T belongs to the genus Stackebrandtia . Whole-cell hydrolysates of strain YIM 64602T contained the amino acid meso-diaminopimelic acid with the sugars mannose, rhamnose and glucose, and a trace of ribose. The major polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine and phosphatidylethanolamine. MK-10(H6), MK-10(H4) and MK-11(H4) were the predominant components in the quinone system. The fatty-acid pattern was mainly composed of the saturated branched-chain acids iso-C16 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C17 : 0. The DNA G+C content was 72.4 mol%. 16S rRNA gene sequence analysis showed the highest pairwise sequence identity (96.0–98.5 %) with the members of the genus Stackebrandtia . Strain YIM 64602T displayed a DNA–DNA relatedness of 43.9±0.4 % with the type strain Stackebrandtia albiflava YIM 45751T. Based on evidence from this polyphasic study, strain YIM 64602T ( = BCRC 16954T = DSM 45928T) is considered to represent a novel species of the genus Stackebrandtia , for which the name Stackebrandtia endophytica is proposed.

Four Gram-positive-staining, microaerophilic, non-spore-forming, fructose-6-phosphate phosphoketolase-positive bacterial strains were isolated from a faecal sample of a 5-year-old ring-tailed lemur (Lemur catta). The strains showed a peculiar morphology, resembling a small coiled snake, a ring shape, or forming a little ‘Y’ shape. The isolated strains appeared identical, and LMC 13T was chosen as a representative strain and characterized further. Strain LMC 13T showed an A3β peptidoglycan type, similar to that found in Bifidobacterium longum . The DNA base composition was 57.2 mol% G+C. Almost-complete 16S rRNA, hsp60, rpoB, dnaJ, dnaG, purF, clpC and rpoC gene sequences were obtained, and phylogenetic relationships were determined. Comparative analysis of 16S rRNA gene sequences showed that strain LMC 13T showed the highest similarity to B. longum subsp. suis ATCC 27533T (96.65 %) and Bifidobacterium saguini DSM 23967T (96.64 %). Strain LMC 13T was located in an actinobacterial cluster and was more closely related to the genus Bifidobacterium than to other genera in the Bifidobacteriaceae . On the basis of these results, strain LMC 13T represents a novel species within the genus Bifidobacterium , for which the name Bifidobacterium lemurum sp. nov. is proposed; the type strain is LMC 13T ( = DSM 28807T = JCM 30168T).

One bacterial strain, designated KIS75-12T, isolated from a soil sample collected from Wonsando island located in Boryeong city, Republic of Korea, was characterized as aerobic, Gram-stain-positive, non-flagellated and a short rod. It grew between temperatures of 15–37 °C, pH 4–9 and 0–3.0 % (w/v) NaCl. The 16S rRNA gene analysis showed the strain was moderately related to Jatrophihabitans endophyticus S9-650T (97.7 %) and revealed low sequence similarity (≤94.7 %) with all the other species with validly published names. Its major fatty acid was iso-C16 : 0. The predominant menaquinone of strain KIS75-12T was MK-9(H4). The polar lipids consisted of diphosphatidylglycerol and several small amounts of phosphatidylinositol, aminolipids and glycolipid. The peptidoglycan contained meso-A2pm as diagnostic diamino acid and the peptidoglycan type is A4γ. The genomic DNA G+C content of the type strain was 72.1 mol%. The combined phenotypic, chemotaxonomic and phylogenetic data showed that strain KIS75-12T could be clearly distinguished from the only member of the genus Jatrophihabitans , J. endophyticus . Therefore, the results of this study indicate the existence of a representative of a novel species of the genus Jatrophihabitans , for which we propose the name Jatrophihabitans soli sp. nov., with strain KIS75-12T ( = KACC 17298T = DSM 45908T  =  NBRC 109658T) as the type strain. An emended description of the genus Jatrophihabitans is also given.

A novel actinomycete, A-T 8343T was isolated from a moist evergreen forest soil sample collected in the Trat Province, Thailand. Based on 16S rRNA gene sequence analysis, strain A-T 8343T belonged to the genus Sinosporangium and was closely related to Sinosporangium siamense A-T 1946T (98.81 %) and Sinosporangium album 6014T (98.54 %). The DNA–DNA relatedness values were 21.8–27 % with S. siamense A-T 1946T and 31.1–31.9 % with S. album 6014T, which were significantly below 70 %. The result differentiated A-T 8343T from the closest species. The organism developed spherical sporangia containing non-motile spores on aerial mycelia. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars contained rhamnose, ribose, madurose and glucose. The predominant menaquinones were MK-9(H2) and MK-9(H4). The diagnostic phospholipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxyl-phosphatidylethanolamine, diphosphatidylglycerol, lyso-phosphatidylethanolamine, N-acetylglucosamine-containing phospholipids, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol-mannosides, aminophosphoglycolipid and one unknown phospholipid. The major cellular fatty acids were saturated C16 : 0, iso C16 : 0, unsaturated C16 : 1 and C18 : 1. Following an evaluation of phenotypic, chemotaxonomic and genotypic characteristics, the new isolate is proposed as a representative novel species of the genus Sinosporangium to be named Sinosporangium fuscum sp. nov. The type strain is A-T 8343T ( = BCC 52770T = NBRC 109516T).

Strain SB026T was isolated from Brazilian rainforest soil and its taxonomic position established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological features consistent with its classification in the genus Amycolatopsis and formed a branch in the Amycolatopsis 16S rRNA gene tree together with Amycolatopsis bullii NRRL B-24847T, Amycolatopsis plumensis NRRL B-24324T, Amycolatopsis tolypomycina DSM 44544T and Amycolatopsis vancoresmycina NRRL B-24208T. It was related most closely to A. bullii NRRL B-24847T (99.0 % 16S rRNA gene sequence similarity), but was distinguished from this strain by a low level of DNA–DNA relatedness (~46 %) and discriminatory phenotypic properties. Based on the combined genotypic and phenotypic data, it is proposed that the isolate should be classified in the genus Amycolatopsis as representing a novel species, Amycolatopsis rhabdoformis sp. nov. The type strain is SB026T ( = CBMAI 1694T = CMAA 1285T = NCIMB 14900T).

An endophytic actinobacterial strain RZ36T, isolated from roots of the salt-marsh plant Halimione portulacoides, was subjected to a polyphasic taxonomic characterization. 16S rRNA gene sequence analysis revealed that this strain belonged to the genus Microbacterium . The closest phylogenetic relative was Microbacterium hominis DSM 12509T, with a pairwise 16S rRNA gene sequence similarity of 98.8 %. The DNA–DNA hybridization value between strain RZ36T and M. hominis DSM 12509T was 16 %. The affiliation to the genus Microbacterium was corroborated by phenotypic and chemotaxonomic characteristics. The cell-wall peptidoglycan type was B2β and the diagnostic diamino acid was ornithine. Whole-cell sugars detected were galactose, glucose, rhamnose, ribose and xylose. The major fatty acids were anteiso-C15 : 0 and iso-C16 : 0 and the major menaquinone was MK-11 (64 %). Main polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol and two unknown glycolipids. The genomic DNA G+C content was 69.7 mol%. Thus, on the basis of phenotypic, genotypic and chemotaxonomic data, strain RZ36T is considered to represent a novel species of the genus Microbacterium , for which the name Microbacterium proteolyticum sp. nov. (type strain RZ36T = DSM 27100T = CECT 8356T) is proposed.

A novel actinomycete, designated strain NEAU-FHS4T, was isolated from the root of black false hellebore (Veratrum nigrum L.). Strain NEAU-FHS4T formed single spores with smooth surfaces on substrate mycelium. The novel strain contained meso-diaminopimelic as amino acid of the peptidoglycan and xylose and glucose as whole-cell sugars. The predominant menaquinones were MK-10(H6) and MK-10(H8). Mycolic acids were not detected. The diagnostic phospholipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol. The predominant cellular fatty acids were iso-C16 : 0, C16 : 0, C18 : 0 and anteiso-C17 : 0. Phenotypic and chemotaxonomic analysis showed that the novel isolate had characteristics typical of members of the genus Plantactinospora . 16S rRNA gene sequence analysis also indicated that strain NEAU-FHS4T belonged to the genus Plantactinospora , with highest sequence similarities to Plantactinospora mayteni YIM 61359T (98.88 %) and Plantactinospora endophytica YIM 68255T (98.85 %). The results of DNA–DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the novel strain from the most closely related strains. Based on morphological, chemotaxonomic and phylogenetic data, strain NEAU-FHS4T is considered to represent a novel species of the genus Plantactinospora , for which the name Plantactinospora veratri sp. nov. is proposed. The type strain is NEAU-FHS4T ( = CGMCC 4.7143T = DSM 46718T).

Two strains of species of the genus Marmoricola , designated KIS18-7T and JOS5-1T, were isolated from soil samples in Korea. The 16S rRNA gene sequence of strain KIS18-7T showed highest similarities with Marmoricola scoriae Sco-D01T (97.8 %), Marmoricola aequoreus SST-45T (97.6 %) and Marmoricola aurantiacus BC 361T (97.3 %), and strain JOS5-1T had highest sequence similarities with M. aequoreus SST-45T (97.5 %) and Marmoricola bigeumensis MSL-05T (97.3 %). The sequence similarity between KIS18-7T and JOS5-1T was 98.1 %. Phylogenetic analysis showed that these strains grouped with species of the genus Marmoricola . The major fatty acids of strain KIS18-7T were iso-C16 : 0 and C17 : 1ω8c, and C17 : 1ω8c, C18 : 0 10-methyl (TBSA), C18 : 1ω9c, C17 : 0 10-methyl and C16 : 0 2-OH for strain JOS5-1T. Strain KIS18-7T contained the polar lipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylcholine, while strain JOS5-1T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, one unknown aminolipid and two unknown phospholipids. The peptidoglycan of both strains contained ll-diaminopimelic acid as the diagnostic diamino acid and a single glycine residue as the interpeptide bridge (type A3γ). The major menaquinone of both strains was MK-8(H4). The G+C contents of the DNA of strains KIS18-7T and JOS5-1T were 68.0 mol% and 62.9 mol%, respectively. These data demonstrate that strains KIS18-7T and JOS5-1T are representatives of two novel species of the genus Marmoricola , for which the names Marmoricola solisilvae sp. nov. (type strain KIS18-7T = KACC 17307T = DSM 27140T = NBRC 109601T) and Marmoricola terrae sp. nov. (type strain JOS5-1T = KACC 17308T = DSM 27141T = NBRC 109602T) are proposed.

The taxonomic position of an endophytic actinomycete, strain GKU 157T, isolated from the roots of a jambolan plum tree (Syzygium cumini L. Skeels) collected at Khao Khitchakut National Park, Chantaburi province, Thailand, was determined using a polyphasic taxonomic approach. 16S rRNA gene sequence analysis revealed that strain GKU 157T belongs to the genus Actinomadura and formed a distinct phyletic line with Actinomadura chibensis NBRC 106107T (98.6 % similarity). Strain GKU 157T formed an extensively branched, non-fragmenting substrate mycelium and aerial hyphae that differentiated into hooked to short spiral chains of about 20 non-motile spores with a warty surface. The cell wall contained meso-diaminopimelic acid and the whole-cell sugars were galactose, glucose, madurose, mannose and ribose. The N-acyl type of muramic acid was acetyl. Mycolic acids were absent. The phospholipids included phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylinositol (PI), phosphatidylinositolmannoside (PIM) and two unknown phospholipids (PLs). The major menaquinone was MK-9(H6) and the predominant fatty acids were C16:0, iso-C16:0, C18:1ω9c, C18:0 and 10-methyl C18:0 (tuberculostearic acid). The genomic DNA G+C content was 73.1 mol%. A combination of DNA–DNA hybridization results and significant differences from related species in cultural, physiological and chemical characteristics indicated that strain GKU 157T represents a novel species of the genus Actinomadura, for which the name Actinomadura syzygii sp. nov. is proposed. The type strain is GKU 157T ( = BCC 70456T = NBRC 110399T).

A moderately halophilic actinomycete strain, designated AH97T, was isolated from Saharan soil in the Hoggar region (south Algeria) and was subjected to polyphasic taxonomic characterization. The morphological and chemotaxonomic characteristics of the strain were consistent with those of the genus Actinoalloteichus. Results of 16S rRNA gene sequence comparison revealed that strain AH97T shared the highest degree of 16S rRNA gene sequence similarity with Actinoalloteichus hymeniacidonis DSM 45092T (99.3 %) and Actinoalloteichus nanshanensis DSM 45655T (98.7 %). However, DNA–DNA hybridization studies showed only 26.5 % relatedness with A. hymeniacidonis DSM 45092T and 28.0 % with A. nanshanensis DSM 45655T. The genotypic and phenotypic data showed that strain AH97T represents a novel species of the genus Actinoalloteichus, for which the name Actinoalloteichus hoggarensis sp. nov. is proposed, with AH97T ( = DSM 45943T = CECT 8639T) as the type strain.

A thermophilic, anaerobic, iron-reducing bacterium (strain DY22619T) was isolated from a sulfide sample collected from an East Pacific Ocean hydrothermal field at a depth of 2901 m. Cells were Gram-stain-negative, motile rods (2–10 µm in length, 0.5 µm in width) with multiple peritrichous flagella. The strain grew at 40–70 °C inclusive (optimum 60 °C), at pH 4.5–8.5 inclusive (optimum pH 7.0) and with sea salts concentrations of 1–10 % (w/v) (optimum 3 % sea salts) and NaCl concentrations of 1.5–5.0 % (w/v) (optimum 2.5 % NaCl). Under optimal growth conditions, the generation time was around 55 min. The isolate was an obligate chemoorganoheterotroph, utilizing complex organic compounds, amino acids, carbohydrates and organic acids including peptone, tryptone, beef extract, yeast extract, alanine, glutamate, methionine, threonine, fructose, mannose, galactose, glucose, palatinose, rhamnose, turanose, gentiobiose, xylose, sorbose, pyruvate, tartaric acid, α-ketobutyric acid, α-ketovaleric acid, galacturonic acid and glucosaminic acid. Strain DY22619T was strictly anaerobic and facultatively dependent on various forms of Fe(III) as an electron acceptor: insoluble forms and soluble forms. It did not reduce sulfite, sulfate, thiosulfate or nitrate. The genomic DNA G+C content was 29.0 mol%. Phylogenetic 16S rRNA gene sequence analyses revealed that the closest relative of strain DY22619T was Caloranaerobacter azorensis MV1087T, sharing 97.41 % 16S rRNA gene sequence similarity. On the basis of physiological distinctness and phylogenetic distance, the isolate is considered to represent a novel species of the genus Caloranaerobacter , for which the name Caloranaerobacter http://dx.doi.org/10.1601/nm.4081 ferrireducens sp. nov. is proposed. The type strain is DY22619T ( = JCM 19467T = DSM 27799T = MCCC1A06455T).

A Gram-stain-positive, solvent-tolerating, aerobic, rod-shaped bacterium that formed terminal endospores was isolated from the organic solvent acetophenone. The strain, designated JC23T, was oxidase- and catalase-positive. The strain grew in the presence of a wide range of organic solvents with partition coefficients (log p values) between 1 and 4, which are exceptionally toxic to micro-organisms. Based on 16S rRNA gene sequence analysis, strain JC23T was identified as belonging to the genus Lysinibacillus and was most closely related to Lysinibacillus manganicus Mn1-7T (98.5 % similarity), L. massiliensis 440831T (97.2 %) and L. chungkukjangi 2RL3-2T (96.8 %). DNA–DNA relatedness of strain JC23T with the type strains of the closest species was <39 %. Strain JC23T grew chemo-organoheterotrophically with optimal growth at pH 7 (range pH 6–9) and at 35 °C (range 25–40 °C). The DNA G+C content was 41 mol%. Major cellular fatty acids of strain JC23T were iso-C15 : 0, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The cell-wall peptidoglycan type was determined to be A4α (l-Lys–d-Asp), which is in agreement with the cell-wall characteristics of the genus Lysinibacillus . The predominant quinone system was MK-7. Polar lipids of strain JC23T included diphosphatidylglycerol, phosphatidylglycerol, two unidentified glycolipids, β-gentiobiosyldiacylglycerol, two unidentified phospholipids and two unidentified lipids. On the basis of our morphological, physiological, genetic, phylogenetic and chemotaxonomic analyses, we conclude that strain JC23T should be assigned to a novel species of the genus Lysinibacillus , for which the name Lysinibacillus acetophenoni sp. nov. is proposed. The type strain is strain JC23T ( = CCUG 57911T = KCTC 13605T = NBRC 105754T = DSM 23394T).

A Gram-stain-positive, endospore-forming, moderately halophilic bacterial strain, NEAU-ST10-40T, was isolated from a saline and alkaline soil in Anda City, China. It was strictly aerobic, rod-shaped and motile by peritrichous flagella. It formed light yellow colonies and grew at NaCl concentrations of 3–15 % (w/v) (optimum, 8 %, w/v), at pH 7.0–9.0 (optimum, pH 8.0) and at 4–60 °C (optimum, 30 °C). It contained meso-diaminopimelic acid in the cell-wall peptidoglycan. Phylogenetic analysis based on 16S rRNA gene sequences indicated that it belonged to the genus Halobacillus. Levels of 16S rRNA gene sequence similarity between strain NEAU-ST10-40T and the type strains of related species of the genus Halobacillus ranged from 98.8 % (Halobacillus alkaliphilus FP5T) to 97.1 % (Halobacillus kuroshimensis IS-Hb7T). DNA–DNA hybridization relatedness values between strain NEAU-ST10-40T and H. alkaliphilus DSM 18525T, Halobacillus campisalis KCTC 13144T, Halobacillus yeomjeoni DSM 17110T, Halobacillus halophilus DSM 2266T, Halobacillus litoralis DSM 10405T, Halobacillus dabanensis DSM 18199T, Halobacillus salinus DSM 18897T, Halobacillus naozhouensis DSM 21183T, Halobacillus trueperi DSM 10404T and Halobacillus salsuginis DSM 21185T were from 43 ± 1 to 19 ± 1 % (mean ± sd). The DNA G+C content was 39.3 mol%. The major fatty acids (>10 %) were anteiso-C15:0, anteiso-C17:0 and iso-C16:0, the only respiratory quinone detected was MK-7, and polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unknown phospholipids and three unknown lipids. On the basis of the data presented, strain NEAU-ST10-40T is considered to represent a novel species, for which the name Halobacillus andaensis sp. nov. is proposed. The type strain is NEAU-ST10-40T ( = CGMCC 1.12153T = DSM 25866T).

A Gram-stain-positive, aerobic, non-motile, rod-shaped, yellow-pigment-producing bacterium, (designated strain CC-CZW004T), was isolated from seafood samples (sea urchins) at Penghu Island in Taiwan. Strain CC-CZW004T grew optimally at pH 7.0 and 30 °C in the presence of 3 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA genes revealed a distinct taxonomic position attained by strain CC-CZW004T with respect to other members of the genus Nocardioides. The novel strain shared highest 16S rRNA gene sequence similarities to Nocardioides daejeonensis JCM 16922T (96.4 %), Nocardioides pacificus JCM 19260T (96.3 %), and Marmoricola scoriae JCM 17444T (96.2 %). The major fatty acids of strain CC-CZW004T consisted of C17:0, C16:1ω5c, C17:1ω8c`, iso-C16:0 and C19:1ω11c/C19:1ω9c (summed feature 6). The diagnostic diamino acid in the cell wall was ll-2,6-diaminopimelic acid. The polar lipid profile was composed of major amounts of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and aminophospholipid. The DNA G+C content was 73.3 mol%. The predominant quinone system was menaquinone (MK-8). On the basis of polyphasic taxonomic evidences presented here, strain CC-CZW004T represents a novel species of the genus Nocardioides, for which the name Nocardioides echinoideorum sp. nov. is proposed. The type strain is CC-CZW004T ( = BCRC 16974T = JCM 30276T).

Using a polyphasic taxonomic approach, we investigated three bacterial strains – IWT30T, IWT8 and IWT75 – isolated from total mixed ration silage prepared in Hachimantai, Iwate, Japan. The isolates comprised Gram-stain positive, non-motile, non-spore-forming, catalase-negative, rod-shaped bacteria. Good growth occurred at 15–45 °C and at pH 4.0–7.5. Their major cellular fatty acids were C18:1ω9c and C19:1 cyclo 9,10.The G+C content of genomic DNA of strain IWT30T was 44.6 mol%. Comparative 16S rRNA gene sequence analysis showed that these novel strains belonged to the genus Lactobacillus. These strains shared 100 % 16S rRNA gene sequence similarity and were most closely related to the type strains of Lactobacillus silagei, Lactobacillus odoratitofui, Lactobacillus similis, Lactobacillus collinoides, Lactobacillus paracollinoides and Lactobacillus kimchicus, with sequence similarity values of 99.5, 98.8, 98.7, 97.8, 97.8 and 96.8 %, respectively. The level of DNA–DNA relatedness between these strains and their closest phylogenetic neighbours was less than 30 %. On the basis of additional phylogenetic analysis of pheS and rpoA gene sequences and phenotypic and chemotaxonomic characteristics, we conclude that these three strains represent a novel species of the genus Lactobacillus, for which we propose the name Lactobacillus mixtipabuli sp. nov. The type strain is IWT30T ( = JCM 19805T = DSM 28580T).

A facultatively anaerobic bacterium, designated strain 1MBB1T, was isolated from basaltic breccia collected from 341 m below the seafloor by seafloor drilling of Rigil Guyot during Integrated Ocean Drilling Program Expedition 330. The cells were straight rods, 0.5 μm wide and 1–3 μm long, that occurred singly and in chains. Strain 1MBB1T stained Gram-positive. Catalase and oxidase were produced. The isolate grew optimally at 30 °C and pH 7.5, and could grow with up to 12 % (w/v) NaCl. The DNA G+C content was 40.5 mol%. The major cellular fatty acids were C16: 1ω11c (26.5 %), anteiso-C15: 0 (19.5 %), C16: 0 (18.7 %) and iso-C15: 0 (10.4 %), and the cell-wall diamino acid was meso-diaminopimelic acid. Endospores of strain 1MBB1T oxidized Mn(II) to Mn(IV), and siderophore production by vegetative cells was positive. Phylogenetic analysis of the 16S rRNA gene indicated that strain 1MBB1T was a member of the family Bacillaceae, with Bacillus foraminis CV53T and Bacillus novalis LMG 21837T being the closest phylogenetic neighbours (96.5 and 96.2 % similarity, respectively). This is the first novel species described from deep subseafloor basaltic crust. On the basis of our polyphasic analysis, we conclude that strain 1MBB1T represents a novel species of the genus Bacillus, for which we propose the name Bacillus rigiliprofundi sp. nov. The type strain is 1MBB1T ( = NCMA B78T = LMG 28275T).

A novel Gram-positive-staining, strictly aerobic and heterotrophic bacterium, designated strain LL-002T, was isolated from organics- and methane-rich seafloor sediment at a depth of 100 m in Kagoshima Bay, Kagoshima, Japan. Colonies were lustreless and translucent white in colour. The temperature, pH and salt concentration ranges for growth were 10–30 °C, pH 6.0–6.5 and 0–1 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that strain LL-002T belongs to the genus Aneurinibacillus of the family Paenibacillaceae. 16S rRNA gene sequence similarities between strain LL-002T and the type strains of species of the genus Aneurinibacillus were 92.8–95.7 %; the highest sequence identity was with the type strain of Aneurinibacillus migulanus. The DNA G+C content of strain LL-002T was 46.2 mol%. MK-7 was the predominant menaquinone. The predominant cellular fatty acids were iso-C15 : 0 and anteiso-C15 : 0, and the cell-wall peptidoglycan contained meso-diaminopimelic acid and glutamic acid, glycine and alanine in addition to muramic acid and glucosamine. The peptidoglycan type was A1γ. In DNA–DNA hybridization assays between strain LL-002T and the type strains of the other species of the genus Aneurinibacillus, the level of hybridization was 6.3–30.1 %. On the basis of its biological features and the 16S rRNA gene sequence comparison presented here, strain LL-002T is considered to represent a novel species of the genus Aneurinibacillus, for which the name Aneurinibacillus tyrosinisolvens sp. nov. is proposed; the type strain is LL-002T ( = NBRC 110097T = CECT 8536T).