Name Bacillus Research Articles

An Experimental Investigation on Mechanical Properties of Bacterial Concrete Using Bacillus Subtilis

Abstract: This Concrete is susceptible to micro crack formation and has pores which are highly undesirable because they provide an open pathway for the ingress of water and other deleterious substances. The use of concrete surface treatments with water proofing materials to prevent the access of aggressive substances is a common way of contributing to concrete durability. However, the most common surface treatments use organic polymers (epoxy, acrylics and polyurethanes) all of which have some degree of toxicity. Bio inspired materials can lead to a more sustainable construction industry, especially when providing new low toxic solutions. Hence, there is an urgent need to pay more attention for improving the properties of concrete with respect to strength and durability, especially in aggressive environments. One effective remedy for closure of cracks is the Bacterial Concrete (BC) which will continuously deposit calcite in concrete. This, phenomenon is called as Microbiologically Induced Calcite Precipitation (MICP). The urease enzyme enables the deposition of calcium carbonate (CaCO3) with the help of bacteria. The bacterial remediation technique surpasses other techniques as it is bio-based, eco-friendly and durable. Bacteria need to offer resistance to withstand high pH of concrete and mechanical stresses during mixing. In these present investigations bacterial sample name Bacillus Subtilis (BS) is considered in concentrations of 104 , 105 and 106 cells/ml of mixing of water. investigations are carried out on strength-related properties such as compressive strength, splitting tensile strength and flexural strength to assess the quality of concrete. The optimum dosage of cell concentration of bacteria is found to be 105 cells/ml for BS based on compressive strength results. M25 grade concrete is designed and used for embedding Bacillus subtilis bacteria.

Bacillus mexicanus sp. nov., a biological control bacterium isolated from the common bean (Phaseolus vulgaris L.) crop in Sinaloa, Mexico.

Strain FSQ1T was isolated from the rhizosphere of the common bean (Phaseolus vulgaris L.) crop sampled in a commercial field located in the Gabriel Leyva Solano community, which belongs to the Guasave municipality (state of Sinaloa, Mexico). Based on its full-length 16S rRNA gene sequence, strain FSQ1T was assigned to the genus Bacillus (100 % similarity). This taxonomic affiliation was supported by its morphological and metabolic traits. Strain FSQ1T was a Gram-stain-positive bacterium with the following characteristics: rod-shaped cells, strictly aerobic, spore forming, catalase positive, reduced nitrate to nitrite, hydrolysed starch and casein, grew in the presence of lysozyme and 2 % NaCl, utilized citrate, grew at pH 6.0-8.0, produced acid from glucose, was unable to produce indoles from tryptophan, and presented biological control against Sclerotinia sclerotiorum. The whole-genome phylogenetic results showed that strain FSQ1T formed an individual clade in comparison with highly related Bacillus species. In addition, the maximum values for average nucleotide identity and from Genome-to-Genome Distance Calculator analysis were 91.57 and 44.20 %, respectively, with Bacillus spizizenii TU-B-10T. Analysis of its fatty acid content showed the ability of strain FSQ1T to produce fatty acids that are not present in closely related Bacillus species, such as C18 : 0 and C20 : 0. Thus, these results provide strong evidence that strain FSQ1T represents a novel species of the genus Bacillus, for which the name Bacillus mexicanus sp. nov. is proposed. The type strain is FSQ1T (CM-CNRG TB51T=LBPCV FSQ1T).

Bacillus changyiensis sp. nov., isolated from coastal sediment.

Two Gram-stain-positive, facultatively anaerobic, motile, endospore-forming, rod-shaped bacteria, designated CLL-3-40T and CLL-7-23, were isolated from coastal sediment sampled in Changyi, Shandong Province, PR China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains were related to the genus Bacillus and close to six type strains of species within the Bacillus licheniformis group. In phenotypic characterization tests, strain CLL-3-40T could grow at 15-50 °C (optimum, 37 °C) and in media with pH 5-9 (optimum pH 7.0), and tolerate up to 12 % (w/v) NaCl. The fermentation broth supernatant extracted by ethyl acetate of strain CLL-3-40T could inhibit aquaculture pathogenic vibrios. The predominant cellular fatty acids of strain CLL-3-40T were anteiso-C15 : 0 (30.7 %) and iso-C15 : 0 (31.5 %); the peptidoglycan from cell-wall contained meso-diaminopimelic acid; the predominant quinone was menaquinone 7; and the major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, an unidentified glycolipid and two unidentified phospholipids. The digital DNA-DNA hybridization values and average nucleotide identities among strains CLL-3-40T and CLL-7-23 and their close type strains were less than 21.9 and 48.4 %, respectively, thereby indicating that strain CLL-3-40T should represent a novel species of the genus Bacillus. The genomic DNA G+C contents were 38.4 mol% in strain CLL-3-40T and 38.3 mol% in strain CLL-7-23. The 12 biosynthetic gene clusters of strain CLL-3-40T were predicted based on results from the online server antiSMASH. Based upon the consensus of phenotypic and genotypic results, strain CLL-3-40T should be classified as representing a novel species of the genus Bacillus, for which the name Bacillus changyiensis sp. nov. is proposed. The type strain is CLL-3-40T (= MCCC 1A14857T=JCM 35755T).

Bacillus arachidis sp. nov., Isolated from Peanut Rhizosphere Soil.

A novel facultatively aerobic bacterium designated SY8 was isolated from a peanut rhizosphere soil sample collected in Jiangsu Province, China. Cells are Gram-stain-positive, rod-shaped, and agar colonies are creamy, opaque, and usually rhizoidal. Strain growth occurs at 30 - 45°C (optimum 30°C), pH 4.0 - 10.0 (optimum pH 6.0) and 0 - 4% (w/v) NaCl (optimum 2%) in Luria-Bertani medium. Phylogenetic analysis of the 16S rRNA gene sequences indicated that strain SY8 forms a distinct lineage in the clade of genus Bacillus and is related to Bacillus pseudomycoides DSM 12442T (99.9%). Phylogenetic analysis of the concatenated gene sequences of 16S rRNA, gryB and rpoD also indicated that strain SY8 forms a distinct lineage in Bacillus. Calculation of the average nucleotide identities and the digital DNA-DNA hybridization values between strain SY8 and the related type Bacillus strains further revealed that strain SY8 represents a distinct species. The predominant cellular fatty acids are iso C15:0 (28.7%) and summed feature 3 (C16:1ω7c and/or C16:1ω6c) (10.3%). The major polar lipids consisted of diphosphatidyl glycerol, phosphatidyl glycerol, phosphatidyl ethanolamine, phosphatidylinositol, and three unidentified phospholipids. The major menaquinone of SY8 was MK-7. Based on phenotypic, phylogenetic, chemotaxonomic, and genomic features, strain SY8 represents a novel species of the genus Bacillus. The name Bacillus arachidis sp. nov. is proposed with strain SY8T (= CCTCC AB 2021100T=LMG 32409T) designated as the type strain.

Isolation of lysozyme producing Bacillus subtilis Strains, identification of the new strain Bacillus subtilis BSN314 with the highest enzyme production capacity and optimization of culture conditions for maximum lysozyme production

• Screening, isolation and characterization of lysozyme-producing Bacillus subtilis strains. • Bacillus subtilis BSN314 was found to have highest biomass production capacity. • Analysis of results was based on two techniques; single factor and orthogonal method. • By single factor, influence of medium composition and growth conditions was determined. Lysozyme is a naturally occurring enzyme that is ubiquitous and plays an important role in innate immunity through its potent bacteriolytic activity. The main objective of this study was to isolate, screen and identify lysozyme-producing Bacillus subtilis strains by 16S rDNA sequencing. Of the four Bacillus subtilis strains isolated (B18, B66, B70, and B124), only one (B66) was selected for further study because it exhibited the comparatively highest enzymatic activity. The 16S rDNA sequence of strain B66 was submitted to the Genbank database with accession number OM327451 and was assigned the name Bacillus subtilis BSN314. Bacillus subtilis BSN314 was selected for further study to maximize biomass production. To obtain the maximum biomass product of Bacillus subtilis BSN314, two statistical methods were applied, one related to the nutritional conditions affecting the growth medium (chemical components), called the single factor optimization method, and the other showing their effects on the cultivation conditions (orthogonal method). The results showed that the composition of the culture medium (carbon, nitrogen, inorganic salts and surfactants) and the culture conditions (temperature, pH, shaking speed and percentage of inoculum) have a specific influence on the biomass production of Bacillus subtilis BSN314. The culture medium containing the combination of A2B3C1D2 with 1.5% glucose, 2.5% soybean peptone, 0.5% K 2 HPO 4 , and 0.1% Tween-20 was determined to be the optimal medium for biomass production whereas, the order of culture media based on the effects of the selected conditions having lysozyme activity is as follows; glucose % > soyapeptone % > Tween-20 % > K 2 HPO 4 %. The results showed that the A2B3C1D2 ratio had the highest enzyme activity of 55 U/mL.

Bacillus fonticola sp. nov., isolated from deep sea cold seep sediment.

In this study, we report a novel Gram-positive bacterium, designated as strain CS13T, isolated from deep-sea sediment collected in the cold seep area of the South China Sea. Growth of strain CS13T occurred at 16-37°C (optimum 25-28°C), pH 7.0-9.0 (optimum, 7.0), and 0-8% (w/v) NaCl (optimum, 2-3%). Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain CS13T belonged to the genus Bacillus. The closest phylogenetic neighbors of strain CS13T are Bacillus carboniphilus JCM 9731T (96.0%), Bacillus pakistanensis NCCP-168T (95.7%) and Bacillus acidicola 105-2T (95.6%). The genomic DNA G + C content of strain CS13T is 43.7mol%. The principal respiratory quinone was menaquinone 7 (MK-7). The polar lipids of CS13T contained diphosphatidylglycerol, phosphatidylglycerol, phospholipid, and glycolipid. The major fatty acids of CS13T contained anteiso-C15:0, anteiso-C17:0, C16:0 and C18:0. Strain CS13T harboured meso-diaminopimelic acid as the diagnostic diamino acid. Phylogenetic, physiological, biochemical, and morphological analyses suggested that strain CS13T represents a novel species of genus Bacillus, and the name Bacillus fonticola sp. nov. is proposed for the type species CS13T (= CCTCC AB 2019194T = JCM 33663T).

Bacillus aerolatus sp. nov., a novel member of the genus Bacillus, isolated from bioaerosols in a school playground.

A Gram-positive, endospore-forming, rod-shaped bacterium with a single flagellum, and a motile strain, designated CX253, was isolated from bioaerosols. The isolate is facultatively anaerobic, is able to grow at 25-45℃ (optimum 37℃) and pH 6.5-10.0 (optimum 7.5), and can tolerate up to 5.0% NaCl (w/v) under aerobic conditions. The diagnostic diamino acid in the cell wall of strain CX253T is meso-diaminopimelic acid, while major isoprenoid quinone is menaquinone 6 (MK-6) along with a smaller amount of MK-7 (20%). The polar lipid profile is composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phospholipids and glycolipids. The major cellular fatty acid is iso-C15:0 and anteiso-C15:0. Phylogenetic analysis based on 16S rRNA gene and genome sequence grouped strain CX253T into the genus Bacillus. The strain was most closely related to Bacillus thermotolerans CCTCC AB 2012108T by comparison of 16S rRNA gene sequence (97.2% similarity) and to Bacillus wudalianchiensis CCTCC AB 2015266T by comparison of gyrB gene sequence (80.1% similarity). The draft genome of strain CX253T comprised 3,929,195bp with a G + C content of 43.3mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain CX253T and phylogenetically related Bacillus species were lower than 95% and 70%, respectively. Thus, the polyphasic evidence generated through phenotypic, chemotaxonomic and genomic methods confirmed that strain CX253T (= GDMCC 1.1608T = KACC 21318T) was a novel species of the genus Bacillus, for which the name Bacillus aerolatus sp. nov. is proposed.

Bacillus tepidiphilus sp. nov., isolated from tepid spring.

A novel Bacillus strain, designated SYSU G01002T, was isolated from a sediment sample collected from tepid spring in Tengchong, Yunnan province, south-west PR China. The 16S rRNA gene sequence analysis showed that the strain SYSU G01002T shared the highest sequence identity with the type strain of Bacillus alkalitolerans (97.7%). Strain SYSU G01002T grew at pH 6.0-8.0 (optimum, pH 7.0), at 28-55°C (optimum, 45°C) and in the presence of 0-2.5% (w/v) NaCl (optimum in the absence of NaCl). It contained meso-2,6-diaminopimelic acid as the cell-wall diamino acid and MK-7 as isoprenoid quinone. The major cellular fatty acids were iso-C15:0, iso-C17:0 and C16:0. The polar were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and unidentified phospholipid. The genomic DNA G + C content was 38.0mol %. The digital DNA-DNA hybridization and average nucleotide identity values between SYSU G01002T and closely related members of the genus Bacillus were below the cut-off level recommended for interspecies identity. Based on the above results, strain SYSU G01002T represents a novel species of the genus Bacillus, for which the name Bacillus tepidiphilus sp. nov. is proposed. The type strain, SYSU G01002T (= KCTC 43131T = CGMCC 1.17491T).

Bacillus pasinlerensis sp. nov., a thermophilic bacterium isolated from a hot spring in Turkey.

A Gram-reaction-positive, endospore-forming bacterium, designated strain P1T, was isolated from water samples collected from Pasinler Hot Spring and characterized using a polyphasic approach to clarify its taxonomic position. Strain P1T was found to have chemotaxonomic and morphological characteristics consistent with its classification in the genus Bacillus. The strain shared the highest 16S rRNA gene sequence identity values with Bacillus thermolactis R-6488T (97.6 %) and Bacillus kokeshiiformis MO-04T (97.2 %) and formed a distinct clade with both type strains in the phylogenetic trees based on 16S rRNA gene sequences. Strain P1T could grow optimally at 55 °C and in the presence of 2 % NaCl. The organism was found to contain meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The predominant menaquinone was determined to be MK-7. The major cellular fatty acids were identified as iso-C15 : 0, iso-C17 : 0 and anteiso-C17 : 0. Based upon the consensus of phenotypic and phylogenetic analyses, strain P1T represents a novel species of the genus Bacillus, for which the name Bacillus pasinlerensis sp. nov. is proposed. The type strain is P1T (=DSM 107529T=CECT 9885T=NCCB 100674T).

Bacillus telluris sp. nov. Isolated from Greenhouse Soil in Beijing, China.

A novel Gram-stain-positive, rod-shaped, endospore-forming bacterium, which we designated as strain 03113T, was isolated from greenhouse soil in Beijing, China. Phylogenetic analysis based on 16S rRNA gene sequences showed strain 03113T is in the genus Bacillus and had the highest similarity to Bacillus solani CCTCC AB 2014277T (98.14%). The strain grew at 4 °C–50 °C (optimum 37 °C), with 0–10% (w/v) NaCl (optimum 5%), and in the range of pH 3.0–12.0 (optimum pH 8.0). Menaquinone was identified as MK-7, and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. The main major cellular fatty acids detected were anteiso-C15:0 (51.35%) and iso-C15:0 (11.06%), which are the predominant cellular fatty acids found in all recognized members of the genus Bacillus. The 16S rRNA gene sequence and core-genome analysis, the average nucleotide identity (ANI), and in silico DNA—DNA hybridization (DDH) value between strain 03113T and the most closely related species were 70.5% and 22.6%, respectively, which supported our conclusion that 03113T represented a novel species in the genus Bacillus. We demonstrated that type strain 03113T (=ACCC 03113T=JCM 33017T) was a novel species in the genus Bacillus, and the name Bacillus telluris sp. nov. was proposed. Strain 03113T secreted auxin IAA and carried the nitrogenase iron protein (nifH) gene, which indicated that strain 03113T has the potential to fix nitrogen and promote plant growth. Bacillus telluris sp. nov. 03113T is a potential candidate for the biofertilizers of organic agriculture areas.

Bacillus dafuensis sp. Nov., Isolated from a Forest Soil in China.

A Gram-staining-positive, motile, rod-shaped, aerobic bacterial strain FJAT-25496T was isolated from a forest soil sample collected from Dafu County, Chengdu City, Sichuan Province, China. Strain FJAT-25496T grew at 15-50°C (optimum 35°C), pH 6.0-10.0 (optimum 8.0), and NaCl tolerance of 0-4.0% (w/v) (optimum 0%). The strain contained iso-C15:0 and anteiso-C15:0 as the predominant cellular fatty acids as well as MK-7 as the only menaquinone. The major polar lipids were diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), and phosphatidyl glycerol (PG). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain FJAT-25496T was a member of the genus Bacillus, and most closely related to Bacillus solani FJAT-18043T (97.8%), Bacillus praedii FJAT-25547T (97.8%), Bacillus depressus BZ1T (97.7%), Bacillus oceanisediminis H2T (97.7%), and Bacillus ciccensis 5L6T (97.6%). Based on complete genome comparisons between strain FJAT-25496T and the type strains of its closely related species, the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values were 73.4-76.3% and 21.7-23.2%, respectively. The genomic DNA G + C content of strain FJAT-25496T was 36.9%. On the basis of phenotypic, phylogenetic, chemotaxonomic, and genomic features, strain FJAT-25496T is considered to represent a novel species of the genus Bacillus, for which the name Bacillus dafuensis sp. nov. is proposed. The type strain is FJAT-25496T (= CCTCC AB 2019182T = KCTC 43120T).

Bacillus fungorum sp. nov., a bacterium isolated from spent mushroom substrate.

A facultatively anaerobic, Gram-stain-positive, spore-forming Bacillus strain, 17-SMS-01T, isolated from spent mushroom substrate in the Fangshan District, Beijing, PR China, was initially identified as a Bacillus cereus group species based on 16S rRNA gene sequences. Strain 17-SMS-01T had the highest sequence similarities to Bacillus wiedmannii FSL W8-0169T (99.9 %), Bacillus albus N35-10-2T (99.9 %), Bacillus luti TD41T (99.9 %) and Bacillus proteolyticus TD42T (99.9 %). However, the average nucleotide identity (ANI) and digital DNA-DNA hybridization (DDH) values between strain 17-SMS-01T and the most closely related species were less than the previously proposed cut-off values of 96 % (ANI) and 70 % (DDH) for differentiating species within the genus, suggesting that this strain represents a novel Bacillus group species. The fatty acid profile of strain 17-SMS-01T, which showed a predominance of iso-C15 : 0 and anteiso-C15 : 0, supported the allocation of the strain to the genus Bacillus. The predominant menaquinone was MK-7 (100%). The major polar lipids were diphosphatidylglycerol, phosphatidyl ethanolamine, phosphatidyl glycerol, an unidentified aminophospholiped and unidentified lipids. The DNA G+C content of the novel strain was 35.0 mol%. The results of physiological and biochemical tests also allowed the phenotypic differentiation of strain 17-SMS-01T from the most closely related recognized species. On the basis of the phylogenetic and phenotypic evidence, strain 17-SMS-01T represents a novel Bacillus species, for which the name Bacillus fungorum sp. nov. is proposed. Type strain of the novel species is 17-SMS-01T (=MCCC 1K03483T=KCTC 33949T).

Bacillus natronophilus sp. nov., an alkaliphilic bacterium isolated from a soda lake.

An alkaliphilic, moderately halophilic, heterotrophic, rod-shaped, spore-forming bacterium (M30T) was isolated from a sediment sample collected from a soda lake (Lake Magadi, Tanzania). Strain M30T was strictly aerobic, catalase-positive, oxidase-negative and non-motile. Growth occurred at 12-43 °C (optimum, 25-30 °C), at pH 8.0-12 (optimum, pH 9.5-10) and at salinities of 0.5-15 % (w/v) NaCl (optimum 5 %). It utilized various sugars and organic acids as sole carbon sources and was positive for amylase, cellulase, gelatinase, protease and xylanase activities. The cell-wall peptidoglycan contained meso-diaminopimelic acid and the polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified lipid and one unidentified phospholipid. The DNA G+C content was 48.9 mol%. The predominant menaquinone was MK-7 and the major fatty acids (>10 %) comprised anteiso-C15 : 0, iso-C15 : 0, and anteiso-C17 : 0. Phylogenetic analysis based on 16S rRNA gene sequence affiliated M30T to the genus Bacillus and showed the highest similarities to Bacillus populi FJAT-45347T (96.4 %) and Bacillus aurantiacus K1-5T (96.2 %). Based on the data from the current polyphasic study, M30T represents a novel species of the genus Bacillus, for which the name Bacillus natronophilus sp. nov. is proposed. The type strain is M30T (=JCM 32118T=CGMCC 1.16739T=MCC 3010T).

Bacillus yapensis sp. nov., a novel piezotolerant bacterium isolated from deep-sea sediment of the Yap Trench, Pacific Ocean.

A Gram-positive, rod-shaped, endospore-forming bacterium with multiple flagella, designated XXST-01T, was isolated from deep-sea sediment of Yap Trench with a depth of 6300m. Activity of oxidase and catalase were found to be positive. Growth was observed at 10-45°C (optimum 37°C), pH 6-9 (optimum 7.0), 0-6% NaCl (optimum 0-0.5%, w/v) and 0.1-50MPa (optimum 0.1MPa). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain XXST-01T belonged to the genus Bacillus. Strain XXST-01T was closely related to Bacillus kyonggiensis NB22T (98.4%), Bacillus siralis 171544T (97.53%), Bacillus massiliosenegalensis JC6T (97.30%), Bacillus oceanisediminis H2T (97.27%), Bacillus mesophilum IITR-54T (97.12%) and Bacillus depressus BZ1T (97.09%). The ANI and the DNA-DNA hybridization estimate values between strain XXST-01T and closely related type strains were 70.91-90.15% and 19.80-40.50%, respectively. The principal fatty acids were iso-C15:0, anteiso-C15:0, iso-C16:0, and iso-C14:0. The G+C content of the chromosomal DNA was 38.2mol%. The respiratory quinone was determined to be MK-7. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid and four unidentified aminophospholipids. The combined genotypic and phenotypic data show that strain XXST-01T represents a novel species within the genus Bacillus, for which the name Bacillus yapensis sp. nov. is proposed, with the type strain XXST-01T (=MCCC 1A14143T = JCM 33181T).

Bacillus taxi sp. nov., a novel endophytic bacterium isolated from root of Taxus chinensis (Pilger) Rehd.

A Gram-staining-positive, aerobic, rod-shaped, endospore-forming bacterium, designated strain M5HDSG1-1T, was originally isolated from a surface-sterilized root of Taxus chinensis (Pilger) Rehd. in Guizhou, PR China. This bacterium was tested by a polyphasic approach to determine its taxonomic position. A 16S rRNA gene-based phylogenetic analysis revealed that M5HDSG1-1T had the greatest similarity to the type strain of Bacillus nealsonii DSM 15077T (99.1 %). The average nucleotide identity values between M5HDSG1-1T and Bacillus nealsonii DSM 15077T and Bacillus circulans NBRC 13626T were 73.3 and 72.8 %, respectively. The digital DNA-DNA hybridization values between M5HDSG1-1T and Bacillus nealsonii DSM 15077T and Bacillus circulans NBRC 13626T were 20.1 and 20.6 %, respectively, which were below the recommended thresholds. M5HDSG1-1T grew at a pH range of 6.0-12.0 (optimum, 7.0-8.0), at temperatures between 10 and 45 °C (optimum, 30 °C) and at 0-2 % (w/v) NaCl (optimum, 1 %). Neither substrate nor aerial mycelia was formed, and no diffusible pigments were observed on the media tested. The predominant isoprenoid quinone was menaquinone-7 (MK-7). The major fatty acids were anteiso-C15 : 0, and iso-C15 : 0. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminophospholipid and an unidentified phospholipid. The DNA G+C content was 37.5 mol%. According to the phylogeneic, phenotypic and chemotaxonomic data, M5HDSG1-1T was clearly distinguishable from other species with validly published names in the genus Bacillus and should therefore be classified as representing a novel species, and we suggest the name Bacillus taxi sp. nov. The type strain is M5HDSG1-1T (=JCM 33117T=CGMCC 1.13668T).

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

A rod-shaped, endospore-forming, aerobic bacterium, designated FJAT-46582T, was isolated from a sediment sample of the coastal region in Xiapu County, Fujian Province in China. Growth was observed at 10-30 °C (optimum, 25 °C), in 0-7.0 % NaCl (0 %) and at pH 6.0-11.0 (pH 8.0), respectively. The cell-wall peptidoglycan contained meso-diaminopimelic acid and the isoprenoid quinone was MK-7. The main fatty acids were anteiso-C17 : 0 (26.5 %), anteiso-C15 : 0 (19.6 %), iso-C15 : 0 (14.4 %) and C16 : 0 (10.5 %). The main polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidyl ethanolamine. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain FJAT-46582T with the genus Bacillus, and showed the highest sequence similarity to Bacillus thermotolerans SGZ-8T (97.6 %) and Bacillus ectoinformans (97.1 %). The average nucleotide identity and in silico DNA-DNA hybridization values between strain FJAT-46582T and the most closely related species were 72.3 and 22.9 %, respectively, which were much lower than the thresholds commonly used to define species (96 and 70 %, respectively) indicating that it belonged to a different taxon. The DNA G+C content was 44.2 mol%. The phenotypic characters and taxono-genomics study revealed that strain FJAT-46582T represents a novel Bacillus species, for which the name Bacillus xiapuensis sp. nov. is proposed. The type strain is FJAT-46582T (=JCM 33155=CCTCC AB 2017047T).

Bacillus safensis subsp. osmophilus subsp. nov., isolated from condensed milk, and description of Bacillus safensis subsp. safensis subsp. nov.

A bacterial strain, designated BC09T, was isolated from a contaminated sample of condensed milk. Phylogenetic analyses based on 16S rRNA gene sequences placed strain BC09T into the genus Bacillus with its closest relatives being Bacillus safensis and Bacillus australimaris with 100 and 99.9 % similarity, respectively. Analysis of the gyrB gene confirmed the closeness of strain BC09T with respect to the species B. safensis since it presented 97.8 and 95.2 % similarity values, respectively, to the type strains of B. safensis and B. australimaris. DNA-DNA hybridization confirmed these results showing averages of 67 and 56 %, respectively, between strain BC09T and the type strains of B. safensis and B. australimaris. Average nucleotide identity blast values obtained for BC09T compared to the closest relative type strains were 95.7 and 67.6 %, respectively, and predicted DNA-DNA hybridization values were 93.1 and 51.9 %, respectively. However, strain BC09T differs from the type strains of its closest relatives in several phenotypic characteristics. MK-7 was the only menaquinone detected and iso-C15:0 and anteiso-C15:0 were the major fatty acids. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, two unidentified phospholipids, two unidentifed glycolipids, three unidentified lipids and one unidentifed phosphoglycolipid. Meso-diaminopimelic acid was detected in the peptidoglycan. The G+C content was 40.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain BC09T represents a new subspecies of B. safensis, for which the name Bacillus safensis subsp. osmophilus subsp. nov. is proposed. The type strain is BC09T (=LMG 30124T, =CECT 9344T).

Bacillus ferrooxidans sp. nov., an iron(II)-oxidizing bacterium isolated from paddy soil.

An endospore-forming bacterium, designated YT-3T, was isolated from a paddy soil in Yingtan, Jiangxi, China. Cells of strain YT-3T were Gram-positive, rod-shaped, facultative anaerobic, catalase, and oxidase positive. The optimum growth temperature and pH were 30°C (ranged from 15 to 50°C) and 6.5-7.0 (ranged from 3 to 11), respectively. Analysis of the 16S rRNA gene sequence showed that strain YT-3T was affiliated to the genus Bacillus and displayed the highest similarity to that of Bacillus drentensis JCM 21707T (98.3%), followed by B. ginsengisoli JCM 17335T (97.8%) and B. fumarioli JCM 21708T (97.0%). The similarity of rpoB gene sequence between strain YT-3T and B. drentensis JCM 21707T, B. ginsengisoli JCM 17335T and B. fumarioli JCM 21708T was 80.4%, 81.5%, and 82.1%, respectively. The genomic DNA G + C content was 44.9 mol%. The predominant respiratory quinone was Menaquinone-7, and meso-diaminopimelic acid was present in the peptidoglycan layer of cell wall. The major fatty acids were C15:0 anteiso (36.2%), C14:0 iso (19.6%), C15:0 iso (17.4%), and C16:0 iso (9.8%). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phospholipids, and ammoniac phospholipids. The DNA-DNA hybridization values between isolate YT-3T and B. drentensis (JCM 21707T), B. ginsengisoli (JCM 17335T), and B. fumarioli (JCM 21708T) were 36.3%, 30.3%, and 25.3%, respectively. On the basis of physiological, genetic and biochemical data, strain YT-3T represented a novel species of the genus Bacillus, for which the name Bacillus ferrooxidans sp. nov was proposed. The type strain is YT-3T (= KCTC 33875T = CCTCC AB 2017049T).

Bacillus alkalilacus sp. nov., isolated from a sediment sample from a lake in India.

An aerobic, endospore-forming, haloalkali-tolerant, Gram-stain-positive, motile, rod-shaped bacterium, designated strain AK73T, was isolated from a sediment sample collected from Sambhar lake, Jaipur, Rajasthan, India. Colonies were circular, 1-2 mm in diameter, glossy, smooth, yellowish and convex with an entire margin after 48 h growth on marine agar at pH 9 and 37 °C. Growth occurred at 15-42 °C, 0-10 % (w/v) NaCl and at a pH range of 7-12. Strain AK73T was positive for catalase and arginine dihydrolase 2 activities, hydrolysis of Tweens 20, 40 and 80, and negative for esculinase, caseinase, gelatinase, β-galactosidase, lipase (Tween 60) and urease activities. The fatty acids were dominated by branched iso-, anteiso-, saturated fatty acids with a high abundance of iso-C15 : 0, anteiso-C15 : 0, C16 : 0 and anteiso-C17 : 0; MK-7 was the major menaquinone. The cell-wall peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, one unidentified aminophospholipid, four unidentified phospholipids and three unidentified lipids. The DNA G+C content of strain AK73T was 54 mol%. Analysis based on comparative 16S rRNA gene sequence analysis indicated that Bacillus alcalophilus was the nearest phylogenetic neighbour, with a pair-wise sequence similarity of 96.0 %. Phylogenetic analysis showed that strain AK73T formed a separate lineage but was loosely associated with a peripheral cluster of organisms that contained Bacillus gibsonii, Bacillus murimartini and Bacillus plakortidis with similarity values of 93.6, 93.5 and 93.4 %, respectively. Based on its phenotypic characteristics and on phylogenetic inference, strain AK73T represents a novel species of the genus Bacillus, for which the name Bacillus alkalilacus sp. nov. is proposed. The type strain is AK73T (=JCM 32184T=MTCC 12637T=KCTC 33880T).

Bacillus lacus sp. nov., isolated from a water sample of a salt lake in India.

A strictly aerobic, alkaliphilic, Gram-stain-positive, motile, rod-shaped bacterium, designated strain AK74T, was isolated from a water sample collected from Sambhar salt lake, Rajasthan, India. Colonies were circular, 1.2 mm in diameter, shiny, smooth, whitish and convex with an entire margin after 48 h growth at 37 °C with pH 9.0. Growth occurred at 25-42 °C, 0-4 % (w/v) NaCl and at a pH of 7-12. Strain AK74T was positive for aesculinase, caseinase, lipase activities and negative for oxidase, catalase, amylase, cellulase, DNase, gelatinase and urease activities. The fatty acids were dominated by branched iso-, anteiso- and saturated fatty acids with a high abundance of iso-C15 : 0, anteiso-C15 : 0, C16 : 0 and C16 : 1 and the cell-wall peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. The DNA G+C content of strain AK74T was 51.6 mol%. A blast sequence similarity search based on 16S rRNA gene sequences indicated that Bacillus niabensis, Bacillus idriensisand Bacillus halosaccharovorans were the nearest phylogenetic neighbours, with a pair-wise sequence similarities of 96.6, 96.6 and 96.5%, respectively. Phylogenetic analysis showed that strain AK74T clustered with Bacillus mangrove and together clustered with Bacillus idriensisand Bacillus indicus. Based on its phenotypic characteristics and on phylogenetic inference, strain AK74T represents a novel species of the genus Bacillus, for which the name Bacilluslacus sp. nov. is proposed. The type strain is AK74T (=MTCC 12638T=KCTC 33946T=JCM 32185T).