Gracilibacillus alcaliphilus sp. nov., a facultative alkaliphile isolated from indigo fermentation liquor for dyeing.

A facultatively alkaliphilic, lactic-acid-producing and halophilic strain, designated SG103(T), was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample for dyeing prepared in a laboratory. 16S rRNA gene sequence phylogeny suggested that SG103(T) is a member of the genus Gracilibacillus with the closest relatives being 'Gracilibacillus xinjiangensis' J2 (similarity: 97.06 %), Gracilibacillus thailandensis TP2-8(T) (97.06 %) and Gracilibacillus halotolerans NN(T) (96.87 %). Cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. The strain grew at temperatures between 13 and 48 °C with the optimum at 39 °C. It grew in the range pH 7-10 with the optimum at pH 9. The isoprenoid quinone detected was menaquinone-7 (MK-7) and the DNA G+C content was 41.3 mol%. The whole-cell fatty acid profile mainly (>10 %) consisted of iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. Unlike other reported species of the genus Gracilibacillus, the strain lacked diphosphatidylglycerol as a major polar lipid. DNA-DNA hybridization experiments with strains exhibiting greater than 96.87 % 16S rRNA gene sequence similarity, 'G. xinjiangensis' J2, G. thailandensis TP2-8(T) and G. halotolerans NN(T), revealed 2±4 %, 4±9 % and 3±2 % relatedness, respectively. On the basis of the differences in phenotypic and chemotaxonomic characteristics, and the results of phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from reported species of the genus Gracilibacillus, strain SG103(T) merits classification as a members of a novel species, for which the name Gracilibacillus alcaliphilus sp. nov. is proposed. The type strain is SG103(T) ( = JCM 17253(T) = NCIMB 14683(T)).

Growth-dependent catalase localization in Exiguobacterium oxidotolerans T-2-2T reflected by catalase activity of cells.

A psychrotolerant and H2O2-resistant bacterium, Exiguobacterium oxidotolerans T-2-2(T), exhibits extraordinary H2O2 resistance and produces catalase not only intracellularly but also extracellularly. The intracellular and extracellular catalases exhibited the same enzymatic characteristics, that is, they exhibited the temperature-dependent activity characteristic of a cold-adapted enzyme, their heat stabilities were similar to those of mesophilic enzymes and very high catalytic intensity. In addition, catalase gene analysis indicated that the bacterium possessed the sole clade 1 catalase gene corresponding to intracellular catalase. Hence, intracellular catalase is secreted into the extracellular space. In addition to intracellular and extracellular catalases, the inner circumference of the cells showed the localization of catalase in the mid-stationary growth phase, which was observed by immunoelectron microscopy using an antibody against the intracellular catalase of the strain. The cells demonstrated higher catalase activity in the mid-stationary growth phase than in the exponential growth phase. The catalase localized in the inner circumference can be dissociated by treatment with Tween 60. Thus, the localized catalase is not tightly bound to the inner circumference of the cells and may play a role in the oxidative defense of the cells under low metabolic state.

Long-term biopersistence of tangled oxidized carbon nanotubes inside and outside macrophages in rat subcutaneous tissue.

Because of their mechanical strength, chemical stability, and low molecular weight, carbon nanotubes (CNTs) are attractive biological implant materials. Biomaterials are typically implanted into subcutaneous tissue or bone; however, the long-term biopersistence of CNTs in these tissues is unknown. Here, tangled oxidized multi-walled CNTs (t-ox-MWCNTs) were implanted into rat subcutaneous tissues and structural changes in the t-ox-MWCNTs located inside and outside of macrophages were studied for 2 years post-implantation. The majority of the large agglomerates were present in the intercellular space, maintained a layered structure, and did not undergo degradation. By contrast, small agglomerates were found inside macrophages, where they were gradually degraded in lysosomes. None of the rats displayed symptoms of cancer or severe inflammatory reactions such as necrosis. These results indicate that t-ox-MWCNTs have high biopersistence and do not evoke adverse events in rat subcutaneous tissue in vivo, demonstrating their potential utility as implantable biomaterials.

Oceanobacillus polygoni sp. nov., a facultatively alkaliphile isolated from indigo fermentation fluid.

A facultatively alkaliphilic, lactic-acid-producing and halophilic strain, designated SA9(T), was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample prepared in a laboratory. The 16S rRNA gene sequence phylogeny suggested that strain SA9(T) was a member of the genus Oceanobacillus with the closest relative being Oceanobacillus profundus KCCM 42318(T) (99.3% 16S rRNA gene sequence similarity). Cells of strain SA9(T) stained Gram-positive and were facultative anaerobic straight rods that were motile by peritrichous flagella. The strain grew between 5 and 48 °C (optimum, 35 °C) and at pH 7-12 (optimum, pH 9). The isoprenoid quinone detected was menaquinone-7 (MK-7) and the DNA G+C content was 40.6 ± 0.9 mol%. The whole-cell fatty acid profile mainly consisted of iso-C(15:0), anteiso-C(15:0), C(16:0) and anteiso-C(17:0). DNA-DNA hybridization with Oceanobacillus profundus DSM 18246(T) revealed a DNA-DNA relatedness value of 23 ± 2%. On the basis of the differences in phenotypic and chemotaxonomic characteristics, and the results of phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from recognized species of the genus Oceanobacillus, strain SA9(T) merits classification as a representative of a novel species of the genus Oceanobacillus, for which the name Oceanobacillus polygoni sp. nov. is proposed. The type strain is SA9(T) ( =JCM 17252(T) =NCIMB 14684(T)). An emended description of the genus Oceanobacillus is also provided.

Amphibacillus indicireducens sp. nov., an alkaliphile that reduces an indigo dye.

Two indigo-reducing alkaliphilic strains, designated strain C40(T) and strain N214, were isolated from a fermented Polygonum Indigo (Polygonum tinctorium Lour.) liquor sample aged for 10 months and obtained from Date City, Hokkaido, Japan. 16S rRNA gene sequence phylogeny suggested that strains C40(T) and N214 were members of the genus Amphibacillus with the closest relative being Amphibacillus xylanus JCM 7361(T) (97.5 % 16S rRNA gene sequence similarity with strain C40(T)), which is the only strain having a 16S rRNA gene sequence similarity higher than 97 % with strain C40(T). Cells of strain C40(T) were Gram-stain-positive, facultatively anaerobic, straight rods that were motile by means of peritrichous flagella. The strains grew between 17 and 39 °C (optimum, 35 °C) and in the pH range of 9.0-12.0. No isoprenoid quinone was detected and the DNA G+C content was 37.5-37.7 mol%. The whole-cell fatty acid profile mainly consisted of iso-C(15 : 0) and anteiso-C(15 : 0). DNA-DNA hybridization of strain C40(T) with Amphibacillus xylanus JCM 7361(T) revealed a DNA-DNA relatedness value of 10±3 %. Owing to the differences in phenotypic characteristics and phylogenetic analyses based on 16S rRNA gene sequences, as well as DNA-DNA relatedness data from reported species of the genus Amphibacillus, the isolates merit classification as a novel species in the genus Amphibacillus, for which the name Amphibacillus indicireducens sp. nov. is proposed. The type strain is C40(T) ( = JCM 17250(T) = NCIMB 14686(T)). An additional strain of the species is N214. An emended description of the genus Amphibacillus is provided.

Oceanobacillus indicireducens sp. nov., a facultative alkaliphile that reduces an indigo dye.

An indigo-reducing facultatively alkaliphilic and halophilic strain, designated strain A21(T), was isolated from a fermented Polygonum indigo (Polygonum tinctorium Lour.) liquor sample aged for 4 days prepared in a laboratory. 16S rRNA gene sequence phylogeny suggested that strain A21(T) was a member of the genus Oceanobacillus with the closest relative being the type strain of Oceanobacillus chironomi (similarity: 96.0 %). The cells of the isolate stained Gram-positive and were facultatively anaerobic straight rods that were motile by peritrichous flagella. The strain grew between 18 and 48 °C with optimum growth at 39 °C. It grew in the pH range of 7-12. It hydrolysed casein, gelatin and Tween 20 but not Tweens 40, 60 and 80, starch or DNA. No isoprenoid quinone was detected and the DNA G+C content was 39.7 mol%. The whole-cell fatty acid profile mainly consisted of iso-C15 : 0, anteiso-C15 : 0 and C16 : 0. DNA-DNA hybridization experiments with O. chironomi revealed 13 % relatedness. Owing to the differences in phenotypic and chemotaxonomic characteristics, and phylogenetic analyses based on 16S rRNA gene sequences and DNA-DNA relatedness data from reported Oceanobacillus species, the isolate merits classification as a representative of a novel species, for which the name Oceanobacillus indicireducens sp. nov. is proposed. The type strain is A21(T) ( = JCM 17251(T)  = NCIMB 14685(T)). The description of the genus Oceanobacillus is also emended.

The effect of enamel matrix derivative on spreading, proliferation, and differentiation of osteoblasts cultured on zirconia.

PURPOSE: This study investigated the effect of enamel matrix derivative (EMD) on spreading, proliferation, and differentiation of osteoblasts cultured on zirconia disks with smooth and rough surfaces. MATERIALS AND METHODS: EMD was added to the culture medium or coated on zirconia disks that had machined (smooth) or sandblasted (rough) surfaces. The effects of EMD on cell proliferation of MC3T3-E1 osteoblastic cells were examined using a hemocytometer. Osteoblastic differentiation was examined by histologic analysis of alkaline phosphatase (ALP) activity and the degree of mineralization. ALP activity was also measured quantitatively. Scanning electron microscopic analysis was performed to observe cell morphology. Enzyme-linked immunosorbent assay of osteocalcin and reverse-transcriptase polymerase chain reaction of osteocalcin, osteopontin, and type 1 collagen were performed to investigate the expression of osteoblast-related genes. RESULTS: The addition of EMD to the medium enhanced the spreading, proliferation, and differentiation of osteoblasts cultured on zirconia. However, when it was coated on zirconia, EMD reduced osteoblastic spreading and adhesion in the early stage of culture, although it enhanced proliferation and differentiation of osteoblasts in later stages. A promotive effect of EMD on osteocalcin mRNA expression, mineralization, and ALP activity of osteoblasts cultured on the rough surface was observed. CONCLUSIONS: EMD may contribute to treatment with zirconia implants via its promotion of osteoblastic proliferation and activity. However, the procedure for application of EMD may be a crucial factor for the outcome of implants.

Brevibacillus nitrificans sp. nov., a nitrifying bacterium isolated from a microbiological agent for enhancing microbial digestion in sewage treatment tanks.

A heterotrophic nitrifying bacterium, designated strain DA2(T), was isolated from a microbiological agent for enhancing microbial digestion in sewage treatment tanks. Cells of strain DA2(T) were Gram-positive, facultatively anaerobic, sporulating rods that were motile by means of peritrichous flagella; they were able to grow at pH 5-8. The major isoprenoid quinone of strain DA2(T) was menaquinone-7 (MK-7) and its cellular fatty acid profile consisted mainly of iso-C(15 : 0) (18.6 %) and anteiso-C(15 : 0) (69.1 %). The DNA G+C content was 54.1 mol%. 16S rRNA gene sequence phylogeny suggested that strain DA2(T) is a member of the genus Brevibacillus, with highest sequence similarities (in parentheses) to the type strains of Brevibacillus choshinensis (99.7 %), B. formosus (99.4 %), B. brevis (99.4 %), B. agri (99.0 %), B. reuszeri (98.8 %), B. parabrevis (98.7 %), B. centrosporus (98.6 %), B. limnophilus (97.4 %), B. panacihumi (97.3 %) and B. invocatus (97.3 %). DNA-DNA hybridization showed less than 60 % relatedness between strain DA2(T) and type strains of the most closely related species given above. Given the significant differences in phenotypic and chemotaxonomic characteristics, and phylogenetic analysis based on the 16S rRNA sequence and DNA-DNA relatedness data, the isolate merits classification as a novel species, for which the name Brevibacillus nitrificans is proposed; the type strain of this species is DA2(T) (= JCM 15774(T) = NCIMB 14531(T)).

Pseudomonas toyotomiensis sp. nov., a psychrotolerant facultative alkaliphile that utilizes hydrocarbons.

A psychrotolerant, facultatively alkaliphilic strain, HT-3(T), was isolated from a sample of soil immersed in hot-spring water containing hydrocarbons in Toyotomi, Hokkaido, Japan. 16S rRNA gene sequence-based phylogeny suggested that strain HT-3(T) is a member of the genus Pseudomonas and belongs to the Pseudomonas oleovorans group. Cells of the isolate were Gram-negative, aerobic, straight rods, motile by a single polar flagellum. The strain grew at 4-42 °C, with optimum growth at 35 °C at pH 7, and at pH 6-10. It hydrolysed Tweens 20, 40, 60 and 80, but not casein, gelatin, starch or DNA. Its major isoprenoid quinone was ubiquinone-9 (Q-9) and the DNA G+C content was 65.1 mol%. The whole-cell fatty acid profile consisted mainly of C(16 : 0), C(16 : 1)ω9c and C(18 : 1)ω9c. Phylogenetic analyses based on gyrB, rpoB and rpoD sequences revealed that the isolate could be discriminated from Pseudomonas species that exhibited more than 97 % 16S rRNA gene sequence similarity and phylogenetic neighbours belonging to the P. oleovorans group including the closest relative of the isolate, Pseudomonas alcaliphila. DNA-DNA hybridization with P. alcaliphila AL15-21(T) revealed 51 ± 5 % relatedness. Owing to differences in phenotypic properties and phylogenetic analyses based on multilocus gene sequence analysis and DNA-DNA relatedness data, the isolate merits classification in a novel species, for which the name Pseudomonas toyotomiensis sp. nov. is proposed. The type strain is HT-3(T) ( = JCM 15604(T)  = NCIMB 14511(T)).

Transmission electron microscopic observation of cells cultured on multiwalled carbon nanotube-coated sponges.

The cell structure and interface between cultured cells and a multiwalled carbon nanotube (MWCNT)-coated sponge (MWCNT-coated sponge) were observed by transmission electron microscopy (TEM). Moreover, the atomic structure of MWCNTs that entered the cells was also examined by means of high-resolution TEM (HRTEM). MWCNTs were observed in the cytoplasm, and a few MWCNTs were recognized in the cell nuclei. Those MWCNTs maintained their structure there. Subcellular organelles did not appear to be different from those on the collagen sponge despite the cellular uptake of MWCNTs.

Enhancement of the nitrogen fixation efficiency of genetically-engineered Rhizobium with high catalase activity.

The vktA catalase gene, which had been cloned from Vibrio rumoiensis S-1T having extraordinarily high catalase activity, was introduced into the root nodule bacterium, Rhizobium leguminosarum bv. phaseoli USDA 2676. The catalase activity of the vktA-transformed R. leguminosarum cells (free-living) was three orders in magnitude higher than that of the parent cells and this transformant could grow in a higher concentration of exogenous hydrogen peroxide (H2O2). The vktA-transformant was inoculated to the host plant (Phaseolus vulgaris L.) and the nodulation efficiency was evaluated. The results showed that the nitrogen-fixing activity of nodules was increased 1.7 to 2.3 times as compared to the parent. The levels of H2O2 in nodules formed by the vktA-transformant were decreased by around 73%, while those of leghemoglobins (Lba and Lbb) were increased by 1.2 (Lba) and 2.1 (Lbb) times compared with the parent. These results indicated that the increase of catalase activity in rhizobia could be useful to improve the nitrogen-fixing efficiency of nodules by the reduction of H2O2 content concomitantly with the enhancement of leghemoglobins contents.

3D collagen scaffolds coated with multiwalled carbon nanotubes: initial cell attachment to internal surface.

The cell adhesion in a multiwalled carbon nanotube-coated collagen sponge (MWCNT-coated sponge) was investigated. Immediately after seeding, the cells adhered to the inner surface of the MWCNT-coated sponge and a significantly larger number of cells were observed there than for a pure collagen sponge used as control. On the MWCNT-coated sponge, the cells appeared favorable adhesion and spread in the early stages in the center part of the sponge which cells rarely attached without MWCNT-coating. It was suggested that the physical structure of MWCNTs was effective for initial adhesion of cells from the result of serum-free culture. MWCNT-coating makes the material a suitable three-dimensional scaffold for cell culturing, as opposed to other scaffold systems where such an effect is not seen.

Psychrobacter piscatorii sp. nov., a psychrotolerant bacterium exhibiting high catalase activity isolated from an oxidative environment.

A Gram-negative, non-motile, psychrotolerant bacterium exhibiting high catalase activity, designated strain T-3-2(T), was isolated from a drain of a fish-processing plant. Its catalase activity was 12 000 U (mg protein)(-1), much higher than the activity of the other Psychrobacter strains tested. The strain grew at 0-30 degrees C and in the presence of 0-12 % NaCl. The predominant isoprenoid quinone was ubiquinone-8 (Q-8), and C(16 : 1)omega9c and C(18 : 1)omega9c were the predominant cellular fatty acids. The DNA G+C content of strain T-3-2(T) was 43.9 mol%. 16S rRNA gene sequence phylogeny suggested that strain T-3-2(T) is a member of the genus Psychrobacter, with the closest relatives being the type strains of Psychrobacter nivimaris (99.2 % similarity), P. aquimaris (98.7 %) and P. proteolyticus (98.5 %). DNA-DNA hybridization showed less than 65 % relatedness with these strains. A phylogenetic tree based on gyrB gene sequences was more reliable, with higher bootstrap values than the 16S rRNA gene sequence-based tree. The result also differentiated the isolate from previously reported Psychrobacter species. Owing to the significant differences in phenotypic and chemotaxonomic characteristics and the phylogenetic and DNA-DNA relatedness data, the isolate merits classification within a novel species, for which the name Psychrobacter piscatorii sp. nov. is proposed. The type strain is T-3-2(T) (=JCM 15603(T) =NCIMB 14510(T)).

Low-voltage and high-voltage TEM observations on MWCNTs of rat in vivo.

In the present study, we focused on the optimal conditions for observation of morphology and atomic structure of carbon nanotube (CNT) in vivo by transmission electron microscopy (TEM). Either low-voltage or high-voltage TEMs was chosen for the high-contrast or high-resolution imaging of subcutaneous tissue and the multi-wall CNT (MWCNT). The morphology and structure of each cell organelle were well recognized using the low-voltage TEM at 75 kV. Individual MWCNTs forming the cluster were also visible by the low-voltage TEM. On the contrary, the high-voltage TEM image at 1250 kV shows poor contrast on both the cell organelles and MWCNTs. However, graphene layers of MWCNT were clearly visible in the HRTEM image using the high-voltage TEM. The influence of the surrounding biological tissue can be disregarded by the high-energy electrons due to their weak scattering/absorption effect in the tissue. It was indicated that the usage of the high-voltage TEM is quite effective to the atomic structure analysis of nano-crystalline materials in vivo.

High-resolution electron microscopy of multi-wall carbon nanotubes in the subcutaneous tissue of rats.

The atomic structure of multi-wall carbon nanotubes (MWCNTs) implanted in the subcutaneous tissue of rats was examined by means of high-resolution transmission electron microscopy (HRTEM). Clusters of the MWCNTs implanted in the subcutaneous tissue were well recognized by the TEM observations. It was indicated that some nanotubes were taken in phagocytes after the 1-year implantation. The deterioration of crystalline structure of the nanotubes in phagocytes was shown by the HRTEM observation. It was suggested that the deterioration of the nanotubes was due to the peeling of the outer graphene layers in the phagocytes.

H2O2 tolerance of Vibrio rumoiensis S-1(T) is attributable to the cellular catalase activity.

The extraordinarily high level of H2O2 tolerance of Vibrio rumoiensis strain S-1(T) when compared with the tolerance levels of strain S-4, a probable catalase-deficient derivative of strain S-1(T), was demonstrated by the introduction of 0-100 mM H2O2 during the mid-exponential growth phase. The contribution of catalase to the H2O2 tolerance was also demonstrated by comparing the catalase-deficient mutant Escherichia coli strain UM2 with a UM2 strain, harboring the plasmid pBSsa1, which carried the strain S-1(T) catalase gene vktA. The decomposition rates of 23-25 mM H2O2 that was introduced in the culture fluids of strain S-1(T) and E. coli UM2 harboring pBSsa1 corresponded to the calatase activities of the cells by spectrophotometric measurements. The presence of cell surface catalase was observed by immunoelectron microscopy, using an antibody for intracellular catalase in strain S-1(T). The high level of H2O2 tolerance of strain S-1(T) was attributable to the catalase activity of the cells. Cell surface catalase is considered to contribute to the catalase activity of strain S-1(T) cells.

Acinetobacter sp. strain Ths, a novel psychrotolerant and alkalitolerant bacterium that utilizes hydrocarbon.

A novel psychrotolerant, alkalitolerant bacterium, strain Ths, was isolated from a soil sample immersed in hot spring water containing hydrocarbons and grown on a chemically defined medium containing n-tetradecane as the sole carbon source. The isolate grew at 0 degrees C but not at temperatures higher than 45 degrees C; its optimum growth temperature was 27 degrees C. It grew in the pH range of 7-9. The strain utilized C(13)-C(30) n-alkane and fluorene at pH 9 and 4 degrees C. To our knowledge, this is the first report on the bacterium that utilizes a wide range of hydrocarbons at a high pH and a low temperature. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Ths is closely related to genomic species 6 ATCC 17979 (99.1% similarity), genomic species BJ13/TU14 ATCC 17905 (97.8% similarity), genomic species 9 ATCC 9957 (97.6% similarity) belonging to the genus Acinetobacter and to Acinetobacter calcoaceticus JCM 6842(T) (97.5% similarity). DNA-DNA hybridization revealed that the isolate has 62, 25, 18 and 19% relatedness, respectively, to genomic species 6 ATCC 17979, genomic species BJ13/TU14 ATCC 17905, genomic species 9 ATCC 9957 and A. calcoaceticus, respectively.

Alkalibacterium indicireducens sp. nov., an obligate alkaliphile that reduces indigo dye.

Indigo-reducing, obligately alkaliphilic strains A11T, F11 and F12 were isolated from indigo fermentation liquor obtained from Tokushima Prefecture, Shikoku, Japan. The isolates grew at pH 9.0-12.3, but not at pH 7.0-8.0. The optimum pH range for growth was 9.5-11.5. They were Gram-negative, facultatively anaerobic, rod-shaped strains with peritrichous flagella. The isolates grew in 0-14 % (w/v) NaCl, with optimum growth at 1-11 %. They grew at temperatures of 15-35 degrees C with optimum growth at around 20-30 degrees C. dl-Lactate was the major end product from d-glucose. No quinones were detected. The peptidoglycan type was A4 alpha, l-Lys (l-Orn)-d-Asp. The major cellular fatty acids were C16 : 0, C16 : 17c and C18 : 19c. The DNA G+C contents were 47.0-47.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequence data indicated that the isolates belong to the genus Alkalibacterium. DNA-DNA hybridization revealed low relatedness values between the isolates and the three phylogenetically most closely related species, Alkalibacterium olivapovliticus, Alkalibacterium psychrotolerans and Alkalibacterium iburiense (<41 %). On the basis of phenotypic characteristics, including hydrolysis of cellulose and fermentation of carbohydrates, and chemotaxonomic characteristics, phylogenetic data and DNA-DNA relatedness data, it is concluded that the isolates merit classification as representatives of a novel species of the genus Alkalibacterium, for which the name Alkalibacterium indicireducens sp. nov. is proposed. The type strain of this species is A11T (=JCM 14232T=NCIMB 14253T).

Bacillus polygoni sp. nov., a moderately halophilic, non-motile obligate alkaliphile isolated from indigo balls.

A moderately halophilic, obligate alkaliphile (growth range pH 8-12), designated strain YN-1(T), was isolated from indigo balls obtained from Ibaraki, Japan. The cells of the isolate stained Gram-positive, and were aerobic, non-motile, sporulating rods which grew optimally at pH 9. The strain grew in 3-14% NaCl with optimum growth in 5% NaCl. It hydrolysed casein and Tweens 20, 40 and 60, but not gelatin, starch, DNA or pullulan. Its major isoprenoid quinone was MK-7 and its cellular fatty acid profile mainly consisted of anteiso-C(15:0), anteiso-C(17:0) and anteiso-C(17:1). 16S rRNA phylogeny suggested that strain YN-1(T) was a member of group 7 (alkaliphiles) of the genus Bacillus, with the closest relative being Bacillus clarkii DSM 8720(T) (similarity 99.5%). However, DNA-DNA hybridization showed a low DNA-DNA relatedness (7%) of strain YN-1(T) with B. clarkii DSM 8720(T). Owing to the significant differences in phenotypic and chemotaxonomic characteristics, and phylogenetic and DNA-DNA relatedness data, the isolate merits classification as a new species, for which the name Bacillus polygoni is proposed. The type strain of this species is YN-1(T) (=JCM 14604(T)=NCIMB 14282(T)).