Name Bacillus Research Articles

Bacillus vanillea sp. nov., Isolated from the Cured Vanilla Bean.

A Gram-positive bacterium, designated strain XY18(T), was isolated from a cured vanilla bean in Hainan province, China. Cells were rod-shaped, endospore producing, and peritrichous flagella. Strain XY18(T) grew at salinities of 0-8 % (w/v) NaCl (optimally 1-4 %), pH 4.0-8.0 (optimally 5.0-7.0 %) and temperature range 20-45 °C (optimally 28-35 °C). The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C15:0, iso-C15:0, anteiso-C17:0, and iso-C17:0. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain XY18(T) was a member of the genus Bacillus, and closely related to B. amyloliquefaciens NBRC 15535(T) and B. siamensis PD-A10(T), with 99.1 and 99.2 % sequence similarity, respectively. However, the DNA-DNA hybridization value between strain XY18(T) and B. amyloliquefaciens NBRC 15535(T) was 35.7 %. The genomic DNA G+C content of strain XY18(T) was 46.4 mol%, significantly differed from B. siamensis PD-A10(T) (41.4 %), which was higher than the range of 4 % indicative of species. On the basis of polyphasic taxonomic study, including phenotypic features, chemotaxonomy, and phylogenetic analyses, strain XY18(T) represents a novel species within the genus Bacillus, for which the name Bacillus vanillea sp. nov. is proposed. The type strain is XY18(T) (=CGMCC 8629 = NCCB 100507).

Bacillus cihuensis sp. nov., isolated from rhizosphere soil of a plant in the Cihu area of Taiwan.

A Gram-positive, moderately halotolerant, rod-shaped, spore forming bacterium, designated strain FJAT-14515(T) was isolated from a soil sample in Cihu area, Taoyuan County, Taiwan. The strain grew at 10-35 °C (optimum at 30 °C), pH 5.7-9.0 (optimum at pH 7.0) and at salinities of 0-5 % (w/v) NaCl (optimum at 1 % w/v). The diagnostic diamino acid of the peptidoglycan of the isolated strain was meso-diaminopimelic acid and major respiratory isoprenoid quinone was MK-7. Major cellular fatty acids were anteiso-C15:0 (40.6 %), iso-C15:0 (20.7 %) and the DNA G+C content of strain FJAT-14515(T) was 37.1 mol %. A phylogenetic analysis based on 16S rRNA gene sequences indicated that strain FJAT-14515(T) belongs to the genus Bacillus, and was most closely related to the reference strains of Bacillus muralis DSM 16288(T) (97.6 %) and Bacillus simplex DSM 1321(T) (97.5 %). Levels of DNA-DNA relatedness between strain FJAT-14515(T) and the reference strains of B. muralis DSM 16288(T) and B. simplex DSM 1321(T) were 27.9 % ± 3.32 and 44.1 % ± 0.57, respectively. Therefore, on the basis of phenotypic, chemotaxonomic and genotypic properties, strain FJAT-14515(T) represents a novel species of the genus Bacillus, for which the name Bacillus cihuensis sp. nov. is proposed. The type strain is FJAT-14515(T) (=DSM 25969(T) = CGMCC 1.12697(T)).

Bacillus haikouensis sp. nov., a facultatively anaerobic halotolerant bacterium isolated from a paddy soil.

A Gram-stain positive, rod-shaped, endospore-forming and facultatively anaerobic halotolerant bacterium, designated as C-89(T), was isolated from a paddy field soil in Haikou, Hainan Province, People's Republic of China. Optimal growth was observed at 37 °C and pH 7.0 in the presence of 4% NaCl (w/v). The predominant menaquinone was identified as MK-7, the major cellular fatty acids were identified as anteiso-C(15:0) and iso-C(15:0), and the major cellular polar lipids were identified as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and two unknown phospholipids. The peptidoglycan type was determined to be based on meso-DAP. Based on 16S rRNA gene sequence similarity, the closest phylogenetic relatives were identified as Bacillus vietnamensis JCM 11124(T) (98.8% sequence similarity), Bacillus aquimaris JCM 11545(T) (98.6%) and Bacillus marisflavi JCM 11544(T) (98.5%). The DNA G+C content of strain C-89(T) was determined to be 45.4 mol%. The DNA-DNA relatedness values of strain C-89(T) with its closest relatives were below 18%. Therefore, on the basis of phylogenetic, chemotaxonomic, and phenotypic results, strain C-89(T) can be considered to represent a novel species within the genus Bacillus, for which the name Bacillus haikouensis sp. nov., is proposed. The type strain is C-89(T) (=KCTC 33545(T) = CCTCC AB 2014076(T)).

Thermotolerant Bacillus kokeshiiformis sp. nov. isolated from marine animal resources compost.

A novel Gram-staining-positive, endospore-forming, rod-shaped, facultatively anaerobic, thermotolerant bacterium, designated strain MO-04(T), was isolated from a marine animal resources (MAR) compost. The 16S rRNA gene sequence of strain MO-04(T) showed 99.4% similarity with Bacillus thermolactis R-6488(T), 94.1% similarity with Bacillus thermoamylovorans CNCM I-1378(T), 93.3% similarity with Bacillus humi LMG 22167(T), 93.2% similarity with Bacillus niacini IFO 15566(T) and the similarities with other species were less than 93%. DNA-DNA relatedness between strain MO-04(T) and B. thermolactis DSM 23332(T) was 45%. The DNA G+C content of strain MO-04(T) was 33.4 mol%, comparatively lower than that of B. thermolactis R-6488(T) (35.0 mol%). Strain MO-04(T) grew at 35-61 °C (optimum 50 °C), pH 4.5-9.0 (optimum pH 7.2) and tolerated up to 8.0% (w/v) NaCl (optimum 2%). The MO-04(T) cell wall peptidoglycan type was meso-2,6-diaminopimelic acid, and the major fatty acids were C(16 : 1), C(14 : 1), C(17 : 0) and C(17 : 1). The major polar lipids were represented by diphosphatidylglycerol and phosphatidylglycerol and two unidentified phospholipids. The analysed polyphasic data presented here clearly indicate that the isolate MO-04(T) is considered to represent a novel species within the genus Bacillus for which the name Bacillus kokeshiiformis sp. nov. is proposed. The type strain of B. kokeshiiformis is MO-04(T) ( = JCM 19325(T) = KCTC 33163(T)).

Bacillus aequororis sp. nov., isolated from marine sediment.

The taxonomic position of a bacterium isolated from a marine sediment sample collected from the Bay of Bengal, Kanyakumari coast, India, was analyzed by using a polyphasic taxonomic approach. The isolated strain, designated as M-8(T), had phenotypic characteristics that matched those of the genus Bacillus and it represents a novel species. The diagnostic cell wall amino acid is meso-DAP. The major menaquinone is MK-7, and the strain has a phospholipid pattern of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and unknown glycolipid. The almost-complete 16S rRNA gene sequence (1,450 bases) of the novel strain was compared with those of closely related species and confirmed that the strain belongs to the genus Bacillus. 16S rRNA gene sequence analysis revealed that strain M-8(T) differs from the closely related species Bacillus horikoshi 99.5%, Bacillus halmapalus 98.3%, and Bacillus cohni 97.4%. However, the DNA-DNA hybridization showed that it had genomic relatedness value of 60.7% with B. horikoshi, B. halmapalus (37.6%), and B. cohnii (29.9%). The DNA G+C content of strain M-8(T) is 40.6mol%. Based on the polyphasic data, strain M-8(T) should be recognized as a novel species of the genus Bacillus, for which the name Bacillus aequororis sp. nov. is proposed. The type strain is M-8(T) (=MTCC 11626(T)=JCM 19304(T)).

Bacillus huizhouensis sp. nov., isolated from a paddy field soil.

A Gram-stain positive, facultative aerobic bacterium, designated as strain GSS03(T), was isolated from a paddy field soil. The cells were observed to be endospore forming, rod-shaped and motile with flagella. The organism was found to grow optimally at 35°C at pH 7.0 and in the presence of 1% NaCl. The strain was classified as a novel taxon within the genus Bacillus on the basis of phenotypic and phylogenetic analyses. The closest phylogenetic relatives were identified as Bacillus psychrosaccharolyticus DSM 6(T) (97.61%), Bacillus muralis DSM 16288(T) (97.55%), Bacillus asahii JCM 12112(T) (97.48%), Bacillus simplex DSM 1321(T) (97.48%) and "Bacillus frigoritolerans" DSM 8801(T) (97.38%). The menaquinone was identified as MK-7, the major cellular fatty acid was identified as anteiso-C15:0 and the major cellular polar lipids as phosphatidylethanolamine, phosphatidylmonomethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and three unknown polar lipids. The DNA G+C content was determined to be 40.2mol%. The DNA-DNA relatedness with the closest relatives was below 48%. Therefore, on the basis of all the results, strain GSS03(T) is considered to represent a novel species within the genus Bacillus, for which the name Bacillus huizhouensis sp. nov. is proposed. The type strain is GSS03(T) (=KCTC 33172(T) =CCTCC AB 2013237(T)).

Bacillus daqingensis sp. nov., a halophilic, alkaliphilic bacterium isolated from saline-sodic soil in Daqing, China.

An alkaliphilic, moderately halophilic, bacterium, designated strain X10-1(T), was isolated from saline-alkaline soil in Daqing, Heilongjiang Province, China. Strain X10-1(T) was determined to be a Gram-positive aerobe with rod-shaped cells. The isolate was catalase-positive, oxidase-negative, non-motile, and capable of growth at salinities of 0-16% (w/v) NaCl (optimum, 3%). The pH range for growth was 7.5-11.0 (optimum, pH 10.0). The genomic DNA G+C content was 47.7 mol%. Its major isoprenoid quinone was MK-7 and its cellular fatty acid profile mainly consisted of anteiso-C15:0, anteiso-C17:0, iso-C15:0, C16:0, and iso-C16:0. The peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylglycerol. Phylogenetic analysis based on 16S rRNA gene sequences showed that X10-1(T) is a member of the genus Bacillus, being most closely related to B. saliphilus DSM15402(T) (97.8% similarity) and B. agaradhaerens DSM 8721(T) (96.2%). DNA-DNA relatedness to the type strains of these species was less than 40%. On the basis of the phylogenetic, physiological, and biochemical data, strain X10-1(T) represents a novel species of the genus Bacillus, for which the name Bacillus daqingensis sp. nov. is proposed. The type strain is X10-1(T) (=NBRC 109404(T) = CGMCC 1.12295(T)).

Bacillus cheonanensis sp. nov. isolated from near poultry farm soil [corrected

A novel bacterial strain, designated PFS-5(T), was isolated from the soil environment with feces of a live poultry farm located in Cheonan, Republic of Korea. Strain PFS-5(T) was Gram-staining-positive, motile, strictly aerobic bacterium, rod-shaped, and endospore-forming. The strain contained meso-diaminopimelic acid in their peptidoglycan and MK-7 menaquinone. The major fatty acids were anteiso-C15:0 (44.2%), C16:0 (22.2%), and iso-C15:0 (16.7%). The DNA G+C content was 40.1 mol%. Comparative 16S rRNA gene sequence analysis identified strain PFS-5(T) in the genus Bacillus, exhibiting the highest level of sequence similarity with type strain of B. herbersteinensis D-1,5a(T) (96.9%), B. humi LMG 22167(T) (96.7%), B. alkalitelluris BA288(T) (96.1%), B. litoralis SW-211(T) (96.0%), and B. luteolus YIM93174(T) (95.5%). The major polar lipids of PFS-5(T) were diphosphatidylglycerol and phosphatidylglycerol. On the basis of result from poly-phasic data, strain PFS-5(T) represents a novel species, for which the name Bacillus cheonanensis sp. nov. is proposed (Type strai PFS-5(T) = KACC 17469(T) = JCM19333(T)).

Bacillus oryzaecorticis sp. nov., a moderately halophilic bacterium isolated from rice husks

A Gram-stain-positive, aerobic, endospore-forming, moderately halophilic rod, designated strain R1(T), was isolated from rice husks and subjected to a taxonomic study using a polyphasic approach. Strain R1(T) produced spherical or ellipsoidal endospores at a subterminal position in swollen sporangia, and was catalase- and oxidase-positive. The isolate grew optimally at 37 °C and pH 6.0-7.0, and could grow in the presence of up to 9% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain R1(T) belongs to the genus Bacillus. The closest relatives of strain R1(T) were Bacillus subtilis subsp. subtilis NCIB 3610(T), Bacillus aquimaris TF-12(T), and Bacillus marisflavi TF-11(T), with 16S rRNA gene sequence similarities of 96.0%, 98.4%, and 98.7%, respectively. DNA-DNA relatedness values between the isolate and the reference strains were ≤42±3%. The predominant menaquinones were MK-5 (50%) and MK-7 (50%). The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethanolamine. The major cellular fatty acids were iso-C(15 : 0) (48.6%) and anteiso-C(15 : 0) (20.6%), and the cell-wall diamino acid was meso-diaminopimelic acid. On the basis of 16S rRNA gene sequence analyses and chemotaxonomic and phenotypic characteristics, it is concluded that strain R1(T) represents a novel species of the genus Bacillus, for which we propose the name Bacillus oryzaecorticis sp. nov. The type strain is R1(T) ( = KACC 17217(T) = KCCM 90231(T) = JCM 19602(T)).

Bacillus mesophilum sp. nov., strain IITR-54T, a novel 4-chlorobiphenyl dechlorinating bacterium.

The taxonomic position of a Gram-positive, endospore-forming bacterium isolated from soil sample collected from an industrial site was analyzed by a polyphasic approach. The strain designated as IITR-54T matched most of the phenotypic and chemical characteristics of the genus Bacillus and represents a novel species. It was found to biodegrade 4-chlorobiphenyl through dechlorination and was isolated through enrichment procedure from an aged polychlorinated biphenyl-contaminated soil. Both resting cell assay and growth under aerobic liquid conditions using 4-chlorobiphenyl as sole source of carbon along with 0.01% yeast extract, formation of chloride ions was measured. 16S rRNA (1,489 bases) nucleotide sequence of isolated strain was compared with those of closely related Bacillus type strains and confirmed that the strain belongs to the genus Bacillus. Strain IITR-54T differs from all other species of Bacillus by at least 2.1% at the 16S rRNA level, and the moderately related species are Bacillus oceanisediminis (97.9%) followed by Bacillus infantis (97.7%), Bacillus firmus (97.4%), Bacillus drentensis (97.3%), Bacillus circulans (97.2%), Bacillus soli (97.1%), Bacillus horneckiae (97.1%), Bacillus pocheonensis (97.1%) and Bacillus bataviensis (97.1%), respectively. The cell wall peptidoglycan contained meso-diaminopimelic acid and the major isoprenoid quinone was MK-7. Major fatty acids are iso-C15:0 (32.4%) and anteiso-C15:0 (27.4%). Predominant polar lipids are diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The results of physiological and biochemical tests allowed the genotypic and phenotypic distinctiveness of strain IITR-54T with its phylogenetic relatives and suggest that the strain IITR-54T should be recognized as a novel species, for which the name Bacillus mesophilum sp. nov. is proposed. The type strain is IITR-54T (=MTCC 11060T=JCM 19208T).

Bacillus pakistanensis sp. nov., a halotolerant bacterium isolated from salt mines of the Karak Area in Pakistan.

A rod shaped, non-motile, endospore forming, Gram-stain positive and moderately halotolerant strain, designated as NCCP-168(T), was isolated from salt mines sampled in the Karak district of Khyber Pakhtunkhwa Province in Pakistan. To delineate its taxonomic position, the strain was subjected to polyphasic characterization. Cells of strain NCCP-168(T) can grow at 10-40(○)C (optimum at 30-35(○)C), in a pH range of 5.0-9.0 (optimum at pH 8.0) and in 0-17% (w/v) NaCl on agar medium. The phylogenetic analysis based on the 16S rRNA gene sequence showed that strain NCCP-168(T) belongs to the genus Bacillus with the highest similarity to Bacillus seohaeanensis BH724(T) (97.1%), and less than 97% similarity with other closely related taxa (95.6% with B. subtilis subsp. subtilis NCIB3610(T)). DNA-DNA relatedness between strain NCCP-168(T) and the type strains of closely related species was lower than 30%. Chemotaxonomic data (major menaquinone, MK-7; cell wall peptidoglycan type, A1γ [meso-diaminopimelic acid]; major fatty acids, iso-C15:0 29.9%, anteiso-C15:0 29.3%, iso-C16:0 11.4%, iso-C14:0 8.9% and anteiso-C17:0 7.0%; major polar lipids, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine) support the affiliation of strain NCCP-168(T) with genus Bacillus. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain NCCP-168(T) can be distinguished from the closely related taxa and thus represents a novel species in the genus Bacillus, for which the name Bacillus pakistanensis sp. nov. is proposed, with the type strain NCCP-168(T) (=KCTC 13786(T)=DSM 24834(T)=JCM 18975(T)).

Bacillus alkalicola sp. nov., an alkaliphilic, gram-positive bacterium isolated from Zhabuye Lake in Tibet, China.

A Gram-positive, alkaliphilic bacterium, designated strain Zby6(T), was isolated from Zhabuye Lake in Tibet, China. The strain was able to grow at pH 8.0-11.0 (optimum at pH 10.0), in 0-8 % (w/v) NaCl (optimum at 3 %, w/v) and at 10-45 °C (optimum at 37 °C). Cells of the isolate were facultatively anaerobic and spore-forming rods with polar flagellum. The predominant isoprenoid quinone was MK-7, and its cell wall peptidoglycan contained meso-diaminopimelic acid. The major cellular fatty acids were iso-C(15:0), C(16:0) and anteiso-C(15:0). The major polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethanolamine. The genomic DNA G+C content of the isolate was 38.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Zby6(T) was a member of the genus Bacillus and most closely related to Bacillus cellulosilyticus DSM 2522(T) (97.7 % similarity). The DNA-DNA relatedness value between strain Zby6(T) and B. cellulosilyticus DSM 2522(T) was 59.2 ± 1.8 %. Comparative analysis of genotypic and phenotypic features indicated that strain Zby6(T) represents a novel species of the genus Bacillus, for which the name Bacillus alkalicola sp. nov. is proposed; the type strain is Zby6(T) (=CGMCC 1.10368(T) = JCM 17098(T) = NBRC 107743(T)).

Bacillus solimangrovi sp. nov., isolated from mangrove soil

Two novel bacterial strains, GH2-4T and GH2-5, were isolated from mangrove soil near the seashore of Weno island in Chuuk state, Micronesia, and were characterized by a polyphasic approach. The two strains were strictly aerobic, Gram-staining-positive, motile, endospore-forming rods that were catalase- and oxidase-positive. Colonies were circular, convex, stringy and transparent yellowish (GH2-4T) or opaque whitish (GH2-5). The 16S rRNA gene sequences of the two isolates were identical. The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus kochii WCC 4582T, B. horneckiae DSM 23495T, B. azotoformans LMG 9581T, B. cohnii DSM 6307T and B. halmapalus DSM 8723T (95.6, 95.4, 95.4, 95.2 and 95.2% similarity, respectively). The partial groEL sequence of strain GH2-4T was identical to that of strain GH2-5 and showed <85% similarity to those of the most closely related strains. The isolates grew at pH 5-12 (optimal growth at pH 9), at 10-40 °C (optimum 30-35 °C) and at 0-9% (w/v) NaCl (optimum 1-3% NaCl). The cell-wall peptidoglycan of strains GH2-4T and GH2-5 contained meso-diaminopimelic acid and cell-wall hydrolysates contained ribose as a major sugar. The DNA G+C content was 36 mol%, and DNA-DNA relatedness between the isolates and five related reference strains was 20-24%. Strain GH2-4T exhibited 81% DNA-DNA relatedness with strain GH2-5. The major cellular fatty acids of both strains were iso-C15:0, iso-C16:0, iso-C14:0 and anteiso-C15:0 and the predominant menaquinone was MK-7. On the basis of the evidence from this polyphasic study, strains GH2-4T and GH2-5 (=KCTC 33143=JCM 18995=DSM 27084) represent a novel species of the genus Bacillus, for which the name Bacillus solimangrovi sp. nov. is proposed; the type strain is GH2-4T (=KCTC 33142T=JCM 18994T=DSM 27083T).

Bacillus bingmayongensis sp. nov., isolated from the pit soil of Emperor Qin’s Terra-cotta warriors in China

A Bacillus-like isolate, strain FJAT-13831(T), isolated from the No. 1 pit soil of Emperor Qin's Terra-cotta Warriors in Xi'an City, China, was studied to determine its taxonomic status. Dominant fatty acids of this organism included iso-C15:0, iso-C17:0, C16:0, iso-C13:0, anteiso-C15:0, and iso-C17:1ω5c. Comparative 16S rRNA gene sequence analysis confirmed the affiliation of this isolate to the genus Bacillus and indicated that it was closely related to Bacillus pseudomycoides DSM 12442(T) (99.72 % similarity). A phylogenetic analysis of the gyrB gene sequence similarities exhibited independent clustering of the isolate FJAT-13831(T) and showed 93.8 % (<95 %) sequence similarity with its closest phylogenetic neighbour B. pseudomycoides DSM 12442(T). Separate standing of the strain FJAT-13831(T) was supported by a whole genome-based phylogenetic analysis with an average nucleotide identity value of 91.47 (<95 %) between isolate FJAT-13831(T) and B. pseudomycoides DSM 12442(T) and was consistent with the results of DNA-DNA hybridization (69.1 % relatedness). These findings support the conclusion that the isolate FJAT-13831(T) represents a novel species, for which the name Bacillus bingmayongensis sp. nov. is proposed. The type strain is FJAT-13831(T) (= CGMCC 1.12043(T) = DSM 25427(T)).

Bacillus enclensis sp. nov., isolated from sediment sample

A novel bacterial strain, designated SGD-1123(T) was isolated from Chorao Island, in Goa Province, India. The strain was found to be able to grow at 15-42°C, pH 5-12 and 0-12% (w/v) NaCl. The whole cell hydrolysates were found to contain meso-diaminopimelic acid, galactose and arabinose. The major fatty acids were identified as iso-C15:0 and anteiso-C15:0, MK-7 was identified as the predominant menaquinone and the predominant polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified aminolipid. The genomic DNA G+C content was determined to be 44.6mol%. Phylogenetic analysis based on 16S rRNA gene sequences placed the isolate within the genus Bacillus and further revealed that strain SGD-1123(T) had highest sequence similarity with Bacillus aquimaris, and forms a separate clade with its closest relatives i.e. B. aquimaris, Bacillus vietnamensis and Bacillus marisflavi, with which it shares 94.5, 94.1 and 94.1% similarity respectively. The phylogenetic, chemotaxonomic and phenotypic analyses indicated that strain SGD-1123(T) represents a novel species within the genus Bacillus, for which the name Bacillus enclensis is proposed. The type strain is SGD-1123(T) (NCIM 5450(T)=CCTCC AB 2011125(T)).

Bacillus xiamenensis sp. nov., isolated from intestinal tract contents of a flathead mullet (Mugil cephalus)

A taxonomic study was carried out on strain HYC-10(T), which was isolated from the intestinal tract contents of a flathead mullet, Mugil cephalus, captured from the sea off Xiamen Island, China. The bacterium was observed to be Gram positive, oxidase and catalase positive, rod shaped, and motile by subpolar flagella. The bacterium was found to grow at salinities of 0-12% and at temperatures of 8-45°C. The isolate was found to hydrolyze aesculin and gelatin, but was unable to reduce nitrate to nitrite. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HYC-10(T) belongs to the genus Bacillus, with highest sequence similarity (99.3%) to Bacillus aerophilus 28K(T), Bacillus stratosphericus 41KF2a(T) and Bacillus altitudinis DSM 21631(T), followed by Bacillus safensis DSM 19292(T) (99.5%) and Bacillus pumilus DSM 27(T) (99.5%), while the sequence similarities to others were all below 97.6%. The genomic ANIm values between strain HYC-10(T) and three type strains (B. altitudinis DSM 21631(T), B. safensis DSM 19292(T) and B. pumilus DSM 27(T)) were determined to range from 89.11 to 91.53%. The DNA-DNA hybridization estimate values between strain HYC-10(T) and the three type strains were from 36.60 to 44.00%. The principal fatty acids identified were iso-C15:0 (39.1%), anteiso-C15:0 (22.7%), iso-C17:0 (13.1%), C16:0 (6.1%), anteiso-C17:0 (5.8%) and iso-C16:0 (5.1%). The G+C content of the chromosomal DNA was determined from the draft genome sequence to be 41.3mol%. The respiratory quinone was determined to be MK-7 (100%). Phosphatidylglycerol, diphosphatidylglycerol, aminoglycolipid, two glycolipids and two unknown phospholipids were found to be present. The combined genotypic and phenotypic data show that strain HYC-10(T) represents a novel species of the genus Bacillus, for which the name Bacillus xiamenensis sp. nov. is proposed, with the type strain HYC-10(T) (=CGMCC NO.1.12326(T)=LMG 27143(T)=MCCC 1A00008(T)).

Bacillus thaonhiensis sp. nov., a New Species, was Isolated from the Forest Soil of Kyonggi University by Using a Modified Culture Method

Using a new culture method for unculturable soil bacteria, we discovered a novel species, NHI-38(T), from the forest soil of Kyonggi University campus, South Korea. It was a Gram-positive, rod-shaped, and endospore-forming bacterial strain. It grew over a wide pH range (6.5-9.5), with an optimum range of pH 7-9, and in a wide range of temperatures (15-60°C), with an optimum range of 35-45°C. Growth was possible at 0-2% NaCl concentration, and the optimal range was between 0.5 and 1.5% NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that this new species clustered within the genus Bacillus; it was closely related to "Bacillus abyssalis" SCSIO 15042(T) (98.86%), B. methanolicus NCIMB 13113(T) (95.97%), B. vietnamensis 15-1(T) (95.8%), B. seohaeanensis BH724(T) (95.5%), B. timonensis MM10403188(T) (95.33%), and B. subtilis subsp. subtilis NCIB 3610(T) (94.87%). The main fatty acid components of this bacterium were iso-C15:0 (35.92%), summed feature 3 (C16:1ω7c/C16:1ω6c; 16.92%), and anteiso-C15:0 (14.19%). The predominant quinone in this bacterial strain was MK-7. The polar lipid profile primarily comprised phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. The genomic DNA G+C composition of the isolate was 40.7mol%. The DNA-DNA hybridization results indicated that this strain was distinct from other Bacillus species, the degree of similarity being 50% with "B. abyssalis", 56% with B. methanolicus, 47% with B. vietnamensis, 43% with B. seohaeanensis, 46% with B. timonensis, and 32% with B. subtilis. Based on our results, we regard strain NHI-38(T) as a novel member of the Bacillus genus, and we propose the name Bacillus thaonhiensis (=KACC 17216(T)=KEMB 9005-019(T)=JCM 18863(T)).

Bacillus paraflexus sp. nov., isolated from compost

A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterium capable of growing at 15-42 °C (optimum 30 °C) and at pH 5-11 (optimum pH 7) was isolated from compost. Its taxonomic position was deduced using a polyphasic approach and the strain was designated RC2(T). 16S rRNA gene sequence analysis showed that the isolate belongs to the division Firmicutes, forming a clade within the cluster containing Bacillus flexus IFO 15715(T), and showed highest similarity to B. flexus IFO 15715(T) (98.1 %). The cell wall contained meso-diaminopimelic acid as the diagnostic diamino acid. The major cellular fatty acids of the novel strain were iso-C15:0 (36.83 %), anteiso-C15:0 (49.19 %) and C16:0 (5.19 %). DNA-DNA hybridization between strain RC2(T) and B. flexus DSM 1320(T) showed a level of relatedness of 54.5 %. The polar lipid profile of strain RC2(T) showed the presence of phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine. The predominant isoprenoid quinone was MK-7 and the G+C content of strain RC2(T) was 37.6 mol%. On the basis of phenotypic characteristics, phylogenetic analysis and the results of biochemical and physiological tests, strain RC2(T) was clearly distinguished from closely related members of the genus, and the strain is assigned to a novel species, for which the name Bacillus paraflexus sp. nov. is proposed. The type strain is RC2(T) ( = MTCC 9831(T) = MCC 2100(T) = KCTC 13724(T) = CCM 7754(T)).

Description of Bacillus toyonensis sp. nov., a novel species of the Bacillus cereus group, and pairwise genome comparisons of the species of the group by means of ANI calculations

Strain BCT-7112T was isolated in 1966 in Japan from a survey designed to obtain naturally occurring microorganisms as pure cultures in the laboratory for use as probiotics in animal nutrition. This strain, which was primarily identified as Bacillus cereus var toyoi, has been in use for more than 30 years as the active ingredient of the preparation TOYOCERIN®, an additive for use in animal nutrition (e.g. swine, poultry, cattle, rabbits and aquaculture). Despite the fact that the strain was initially classified as B. cereus, it showed significant genomic differences from the type strains of the B. cereus group that were large enough (ANI values below 92%) to allow it to be considered as a different species within the group. The polyphasic taxonomic study presented here provides sufficient discriminative parameters to classify BCT-7112T as a new species for which the name Bacillus toyonensis sp. nov. is proposed, with BCT-7112T (=CECT 876T; =NCIMB 14858T) being designated as the type strain. In addition, a pairwise comparison between the available genomes of the whole B. cereus group by means of average nucleotide identity (ANI) calculations indicated that besides the eight classified species (including B. toyonensis), additional genomospecies could be detected, and most of them also had ANI values below 94%. ANI values were on the borderline of a species definition only in the cases of representatives of B. cereus versus B. thuringiensis, and B. mycoides and B. weihenstephanensis.

Bacillus songklensis sp. nov., isolated from soil

A Gram-stain-positive, spore-forming, rod-shaped, motile, strictly aerobic bacterial strain, designated CAU 1033(T), was isolated from soil and its taxonomic position was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CAU 1033(T) formed a distinct lineage within the genus Bacillus and was most closely related to Bacillus drentensis KCTC 13025(T) (similarity 95.9 %). CAU 1033(T) contained MK-7 as the only isoprenoid quinone and iso-C15 : 0 and anteiso-C15 : 0 as the major fatty acids. The cell wall peptidoglycan of strain CAU 1033(T) contained meso-diaminopimelic acid and the major whole-cell sugars were arabinose, sucrose and ribose. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids, four unidentified aminophospholipids, an unidentified aminolipid, two unidentified glycolipids and another unidentified polar lipid. The DNA G+C content was 41.4 mol%. On the basis of phenotypic data and phylogenetic inference, strain CAU 1033(T) was classified as a representative of a novel species in the genus Bacillus for which the name Bacillus songklensis sp. nov. is proposed. The type strain is CAU 1033(T) ( = KCTC 13881(T) = CCUG 61889(T)).