Potential osteoporosis-blocker Sparassis crispa polysaccharide: Isolation, purification and structure elucidation.

This study aimed to investigate the structural characterization of water-soluble polysaccharides from Sparassis crispa and their effects on the proliferation and differentiation of mouse osteoblasts. Three fractions (F-1, F-2, and F-3) were obtained from crude polysaccharides by a DEAE-52 cellulose column. The main fraction (F-1) was further purified by polysaccharide gel purification systems to obtain purified water-soluble Sparassis crispa polysaccharide (SCPS). The chemical structure of SCPS was analyzed by gas chromatography, Fourier transform infrared spectroscopy, methylation analysis, and nuclear magnetic resonance spectroscopy. The monosaccharide compositional analysis revealed that SCPS consisted of fucose, arabinose, galactose, glucose, xylose, mannose, ribose, galacturonic acid, glucuronic acid, and mannuronic acid in a molar ratio of 17.37:1.94:25.52:30.83:1.14:0.30:4.98:2.87:2.65. Moreover, the backbone of SCPS was composed of →3)-ß-d-Glcp-(1→4)-ß-d-Glcp-(1→, with side chains attached to the backbone at the O-6 positions through the →3,6)-ß-d-Glcp-(1→ linkage. The in vitro experiments were conducted to investigate the effects of SCPS on the proliferation and differentiation of mouse osteoblasts. The results showed that SCPS significantly enhanced the proliferation and differentiation of mouse osteoblasts, indicating their potential as a pharmaceutical agent for promoting osteoblast proliferation and differentiation.

Halomonas faecis sp. nov., a halophilic bacterium isolated from human faeces.

A novel moderately halophilic, Gram-stain-negative, catalase- and oxidase-positive, strictly aerobic, non-sporulating, non-motile rod, designated strain JSM 104105 T, was isolated from human faeces. Strain JSM 104105 T was able to grow with 0.5-18% (w/v) NaCl (optimum 4-9%), at pH 6-10.5 (optimum pH 7-8) and at 10-40 °C (optimum 30 °C) in complex media. The major cellular fatty acids were C18:1ω7c, C16:0, C16:1ω7c and/or C16:1ω6c, C19:0 cyclo ω8c and C12:0 3-OH. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminophospholipid, an unidentified glycolipid and three unidentified phospholipids. The predominant respiratory quinone was Q-9 and the genomic DNA G + C content was 64.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JSM 104105 T should be assigned to the genus Halomonas, and was most closely related to Halomonas gudaonensis SL014B-69 T (99.0% sequence similarity), followed by Halomonas azerbaijanica TBZ202T (98.6%) and Halomonas lysinitropha 3(2)T (97.3%). The whole genomic analysis showed that strain JSM 104105 T constituted a different taxon separated from the recognized Halomonas species. Combined data from phenotypic and genotypic studies demonstrated that strain JSM 104105 T represents a new species of the genus Halomonas, for which the name Halomonas faecis sp. nov. is proposed. The type strain is JSM 104105 T (= CCTCC AB 2014160 T = CGMCC 1.12945 T = KCTC 42146 T).

Sediminibacillus terrae sp. nov., a moderate halophile isolated from non-saline farm soil.

A Gram-stain-positive, moderately halophilic, strictly aerobic, endospore-forming, rod-shaped bacterium, strain JSM 102062T, was isolated from a non-saline farm soil sample collected from Dehang Canyon in Hunan, PR China. Growth occurred with 0.5-20 % (w/v) NaCl (optimum 4-7 %) at pH 5.5-11.0 (optimum pH 8.0) and at 20-50 °C (optimum 30-35 °C). Contained cell-wall peptidoglycan based on meso-diaminopimelic acid and possessed menaquinone-7 (MK-7) as the major respiratory isoprenoid quinone. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, five unidentified phospholipids and an unidentified glycolipid. The DNA G+C content was 44.1 mol%. Phylogeny based on 16S rRNA gene sequences indicated that strain JSM 102062T belonged to the genus Sediminibacillus, sharing high 16S rRNA gene sequence similarities to Sediminibacillus halophilus EN8dT (99.4 %) and Sediminibacillus albus NHBX5T (98.3 %). The whole genomic analysis showed that strain JSM 102062T constituted a different taxon separated from the recognized Sediminibacillus species. Combined data from phenotypic and genotypic studies demonstrated that strain JSM 102062T represents a noval species of the genus Sediminibacillus, for which the name Sediminibacillus terrae sp. nov. is proposed; the type strain is JSM 102062T (=CCTCC AB 2014166T = CGMCC 1.12957T=DSM 28949T=KCTC 33541T).

Two new guaiane-type sesquiterpenes from Curcuma kwangsiensis and their inhibitory activity of nitric oxide production in lipopolysaccharide-stimulated macrophages.

Two new guaiane-type sesquiterpenes, kwangsiensis A and B (1-2) were isolated from the roots of Curcuma kwangsiensis. Their structures were elucidated by extensive spectroscopic methods, including NMR, circular dichroism (CD) and high-resolution mass-spectrometry. The anti-inflammatory activity of the two compounds was evaluated on the basis of their inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in mouse RAW 264.7 macrophages. Compounds 1 and 2 showed moderate inhibitory activities with IC50 values of 27.4 and 35.1 µM, respectively.

Removal of sulfate and heavy metals by sulfate-reducing bacteria in an expanded granular sludge bed reactor.

The objective of this study is to utilize an expanded granular sludge bed reactor for removing heavy metals and sulfate from synthetic acid-mine drainage (AMD) using sewage sludge fermentation centrate as the substrate. The reactor was bioaugmented with an enriched consortium of sulfate-reducing bacteria (SRB). The bioreactor performance was studied with different chemical oxygen demand [Formula: see text] ratios, liquid upflow velocity (Vup), hydraulic retention time (HRT) and influent pH. The highest COD, sulfate and heavy metal removal efficiencies were achieved at a [Formula: see text] ratio of 1.5, Vup of 4.0 m/h, HRT of 15 h and influent pH of 6.0 (68.2%, 92.1% and 100%, respectively). The activity of SRB greatly increases the effluent pH. Even at an influent pH of 3.0, 60.8% of sulfate, 41.3% of COD and 91.2% of heavy metals could be removed, and the effluent quality can meet the national discharge standard of China. The activity tests demonstrate that the sludge fermentation centrate is an excellent carbon source for SRB. This study shows the potential of synchronous treatment of residual sewage sludge and treatment of AMD.

The new steroidal glycosides from the aerial parts of Lepidogrammitis drymoglossoides.

Two new steroidal glycosides ponasteroside C (1) and ponasteroside D (2) were isolated from the aerial parts of Lepidogrammitis drymoglossoides. Their structures were elucidated by various spectroscopic techniques (IR, HRESIMS, 1D and 2D NMR). All compounds were evaluated for their cytotoxicity against HeLa and HCT-8 cell lines, and compounds 1 and 2 showed mild activity against all the test cell lines.

[Effects of Climatic Factors on Contents of Major Medical Components in Smilax china from Different Regions in Western Hunan].

OBJECTIVE: To study the impact of climatic factors on the major medical component in Smilax china, and to supply a scientific and standard operation protocol on the introduction and cultivation. METHODS: Detect the content of major medical components, such as baicalin and astibin, in Smilax china from eight counties of Xiangxi Autonomous Prefecture. And establish their relationship with annual average temperature, Jan average temp, Jul average temp, valid accumulative temp (≥ 10 degress C), annual maximum temp, annual minimum temp, annual precipitation, annual sunshine amount, non-frost period and relative humidity by using the methods of partial least squares regression analysis (PLS). RESULTS: Relative humidity, annual minimum temp and annual precipitation are the dominant factors. Annual minimum temp, annual average temp and valid accumulative temp were significantly correlated to the content of major medical components, thus, relative humidity, annual precipitation and non-frost period were negatively related to them. CONCLUSION: This study provides a scientific basis for resources protection,introduction and cultivation of Smilax china.

[Analysis of chemical components of volatile oil from Paris polyphylla and their antibacterial activities].

OBJECTIVE: To analyze the chemical components of volatile oil from Paris polyphylla by GC-MS, and to investigate their antimicrobial activities. METHODS: The chemical compositions of essential oil, which from Paris polyphylla obtained by supercritical carbon dioxide, were analyzed by GC-MS, and their antibacterial properties for five common pathogens were screened by filter paper method. RESULTS: The components in the volatile oils were analyzed by GC-MS, and 75 compounds comprising for 98.46% of the essential oil were identified. The major components of essential oil were phthalic acid-isobutyl-hex-3-enyl ester, 9,12-octadecadienoic acid, 2 (1H)-naphthalenone, octahydro-4alpha-methyl-7-(1-methylethyl)-, glycerin, 9-octadecenoic acid-methyl ester, 1,2-benzenedicarboxylic acid-butyl-2-methylpropyl ester, bis(2-ethylhexyl) phthalate, 2-myristynoyl-glycinamide, ethanol, 2-(9,12) -octadecadienyloxy, hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, dodecanoic acid, 10-methyl, methyl ester, diglycerol and 3,3-diethoxy-1-propanol; The bacteriostatic experiment results indicated that this volatile oil had strong inhibitory effects on Micrococcus, Aerobacter and Brevibacterium, whereas it had poor effects Escherichia coli and Proteus vulgaris. CONCLUSION: The experiment provides a scientific basis for further development and utilization of Paris polyphylla.

Rothia marina sp. nov., isolated from an intertidal sediment of the South China Sea.

A novel non-sporulating, non-motile, catalase-positive, oxidase-negative, facultatively anaerobic, Gram-positive coccus, designated strain JSM 078151(T), was isolated from an intertidal sediment sample collected from Naozhou Island in the South China Sea, China. Growth was found to occur in the presence of 0-15 % (w/v) NaCl (optimum 0.5-3 % (w/v) NaCl), at pH 6.5-10.5 (optimum pH 7.0-8.0) and at 5-35 °C (optimum 25-30 °C). The peptidoglycan type was determined to be A3a, containing lysine, glutamic acid and alanine. The major cellular fatty acid identified was anteiso-C15:0 and the predominant menaquinones are MK-7 and MK-8. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylglycerol, glycolipid and one unidentified phospholipid. The genomic DNA G+C content of strain JSM 078151(T) was determined to be 55.2 mol%. A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain JSM 078151(T) should be assigned to the genus Rothia, and was most closely related to Rothia nasimurium CCUG 35957(T) (98.3 % sequence similarity), followed by Rothia amarae J18(T) (97.5 %) and Rothia terrae L-143(T) (97.3 %). A combination of phylogenetic analysis, DNA-DNA relatedness values, phenotypic characteristics and chemotaxonomic data supports the suggestion that strain JSM 078151(T) represents a novel species of the genus Rothia, for which the name Rothia marina sp. nov. is proposed. The type strain is JSM 078151(T) (= DSM 21080(T) = KCTC 19432(T)).

Functional connectivity changes in patients with absence epilepsy studied using resting-state functional MRI.

Functional connectivity is altered in several mental disorders. We used resting-state functional MRI to examine the alterations in functional connectivity that occur in patients with absence epilepsy. We found an altered functional connectivity within and between functional modules in patients with absence epilepsy. Some brain regions had a greater number of altered connections. The functional connectivity within and between modules in absence epilepsy patients showed an increase in the number of positive connections and a decrease in the number of the negative connections. In particular, the superior frontal gyrus demonstrated both an increased number of connections with other nodes of the frontal default mode network and a decreased number of connections with the limbic system. These findings provide a new perspective and shed light on how the balance of connections within and between modules may contribute to the development of absence epilepsy.

Sphingomonas hunanensis sp. nov., isolated from forest soil.

A novel Gram-negative, catalase- and oxidase-positive, strictly aerobic, non spore-forming, rod-shaped bacterium, designated strain JSM 083058(T), was isolated from non-saline forest soil in Hunan Province, China. Growth occurred with 0-8% (w/v) NaCl (optimum, 0.5-3%) at pH 6.0-10.0 (optimum, pH 7.0) and at 5-35°C (optimum, 25-30°C). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JSM 083058(T) fell within the cluster comprising species of the genus Sphingomonas, clustering with Sphingomonas aestuarii K4(T), with which it shared highest 16S rRNA gene sequence similarity (99.2%). The chemotaxonomic properties of strain JSM 083058(T) were consistent with those of the genus Sphingomonas. The predominant respiratory quinone was ubiquinone Q-10, and the major cellular fatty acids were summed feature 8 (C18:1ω7c/C18:1ω6c), C16:0, summed feature 3 (C16:1ω7c/C16:1ω6c) and C17:1ω6c. The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and sphingoglycolipid. The genomic DNA G+C content of strain JSM 083058(T) was 65.5 mol%. The combination of phylogenetic analysis, DNA-DNA relatedness, phenotypic characteristics and chemotaxonomic data supported the view that strain JSM 083058(T) represents a novel species of the genus Sphingomonas, for which the name Sphingomonas hunanensis sp. nov. is proposed. The type strain is JSM 083058(T) (=CCTCC AA 209011(T) = DSM 22213(T)).

Jeotgalicoccus nanhaiensis sp. nov., isolated from intertidal sediment, and emended description of the genus Jeotgalicoccus.

A novel non-sporulating, non-motile, catalase- and oxidase-positive, strictly aerobic, Gram-positive-staining coccus, strain JSM 077023(T), was isolated from an intertidal sediment sample collected from Naozhou Island in the South China Sea, China. Growth occurred in the presence of 0.5-25 % (w/v) NaCl [optimum, 2-5 % (w/v) NaCl] and at pH 5.5-10.5 (optimum, pH 7.0-8.0) and at 4-45 °C (optimum, 30-35 °C). The major amino acid constituents of the cell wall were alanine, glycine and lysine. The major cellular fatty acids were anteiso-C15:0 and iso-C15:0. The strain contained MK-7 and MK-6 as the predominant respiratory quinones and diphosphatidylglycerol, phosphatidylglycerol and an unidentified phospholipid as the polar lipids. The genomic DNA G+C content of strain JSM 077023(T) was 41.3 mol%. A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain JSM 077023(T) should be assigned to the genus Jeotgalicoccus and was most closely related to the type strains of Jeotgalicoccus halotolerans (sequence similarity 99.0 %) and Jeotgalicoccus aerolatus (99.0 %), followed by Jeotgalicoccus coquinae (98.6 %) and Jeotgalicoccus psychrophilus (97.4 %). 16S rRNA gene sequence similarities of less than 97 % were observed with other species of the genus Jeotgalicoccus. Levels of DNA-DNA relatedness between strain JSM 077023(T) and the type strains of J. halotolerans, J. aerolatus, J. coquinae and J. psychrophilus ranged from 36.8 to 22.7 %. The combination of phylogenetic analysis, DNA-DNA relatedness values, phenotypic characteristics and chemotaxonomic data supported the suggestion that strain JSM 077023(T) represents a novel species of the genus Jeotgalicoccus, for which the name Jeotgalicoccus nanhaiensis sp. nov. is proposed. The type strain is JSM 077023(T) ( = DSM 23006(T) = KCTC 13714(T)). An emended description of the genus Jeotgalicoccus is also presented.

Zhihengliuella salsuginis sp. nov., a moderately halophilic actinobacterium from a subterranean brine.

A novel moderately halophilic, alkaliphilic, non-motile, non-sporulating, catalase-positive, oxidase-negative, aerobic, coccus-shaped, Gram-positive bacterium, designated strain JSM 071043(T), was isolated from a subterranean brine sample collected from a salt mine in Hunan Province, China. Growth occurred with 0.5-20% (w/v) NaCl (optimum 5-10%) at pH 6.5-10.5 (optimum pH 8.5) and at 10-40 degrees C (optimum 25-30 degrees C). Good growth also occurred in the presence of 0.5-20% (w/v) KCl (optimum 5-8%) or 0.5-25% (w/v) MgCl(2).6H(2)O (optimum 5-10%). The peptidoglycan type was A4alpha (L: -Lys-L: -Ala-L: -Glu) and major cell-wall sugars were tyvelose and mannose. The major cellular fatty acids were anteiso-C(15:0), iso-C(16:0) and anteiso-C(17:0). Strain JSM 071043(T) contained MK-9 and MK-8 as the predominant menaquinones and diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol as the major polar lipids. The DNA G + C content was 67.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JSM 071043(T) was a member of the suborder Micrococcineae, and was most closely related to Zhihengliuella halotolerans YIM 70185(T) (sequence similarity 98.9%) and Zhihengliuella alba YIM 90734(T) (98.2%), and the three strains formed a distinct branch in the phylogenetic tree. The combination of phylogenetic analysis, DNA-DNA relatedness values, phenotypic characteristics and chemotaxonomic data supports the proposal that strain JSM 071043(T) represents a novel species of the genus Zhihengliuella, for which the name Z. salsuginis sp. nov. is proposed. The type strain is JSM 071043(T) (= DSM 21149(T) = KCTC 19466(T)).

Nocardiopsis terrae sp. nov., a halophilic actinomycete isolated from saline soil.

A Gram-positive, moderately halophilic, facultatively alkaliphilic, catalase- and oxidase-positive, obligately aerobic, filamentous actinomycete strain, designated YIM 90022(T), was isolated from saline soil collected from the Qaidam Basin, north-west China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the new isolate was a member of the genus Nocardiopsis and the sequence similarities between the isolate and the type strains of members of the genus Nocardiopsis were in the range of 95.1-98.7%. Phenotypic and chemotaxonomic properties of this organism also indicated that strain YIM 90022(T) was a member of the genus Nocardiopsis. The strain grew well on most of the media tested, producing yellow-white to deep brown substrate mycelium and white aerial mycelium. Light gray to deep brown diffusible pigments were produced. The substrate mycelium was well developed and fragmented with age; the aerial mycelium produced long, straight to flexuous spore chains with non-motile, smooth-surfaced, rod-shaped spores on them. The strain grew in the presence of 1-15% (w/v) total salts (optimum, 3-5%) and at pH 6.0-10.5 (optimum, pH 8.5) and 10-45 degrees C (optimum, 30 degrees C). Whole-cell hydrolysates of strain YIM 90022(T) contained meso-diaminopimelic acid and no diagnostic sugars. The predominant menaquinones were MK-10(H(4)), MK-9(H(8)), MK-10(H(6)) and MK-10(H(8)). Polar lipids comprised diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylmethylethanolamine. The major cellular fatty acids were iso-C(16:0), anteiso-C(17:0), 10-methyl-C(18:0) and 10-methyl-C(17:0). The DNA G + C content of strain YIM 90022(T) was 71.5 mol%. The combination of phylogenetic analysis, DNA-DNA relatedness data, phenotypic characteristics and chemotaxonomic data supported the suggestion that strain YIM 90022(T) represents a new species of the genus Nocardiopsis, for which the name Nocardiopsis terrae sp. nov. is proposed. The type strain is YIM 90022(T) (=CCTCC AA 208011(T) =KCTC 19431(T)).

[Diversity of halophilic and halotolerant bacteria isolated from non-saline soil collected from Xiaoxi National Natural Reserve, Hunan Province].

OBJECTIVE: To investigate the diversity of cultivable halophilic and halotolerant bacteria isolated from ordinary non-saline soil samples collected from Xiaoxi National Natural Reserve (28 degrees 42' 15"-28 degrees 53' 15" N, 110 degrees 6' 50"-110 degrees 21' 35" E), Hunan Province, China. METHODS: Bacterial strains were isolated from the samples by using the conventional culture-dependent method and investigated by using phylogenetic analysis based on 16S rRNA gene sequence comparisons. RESULTS: We isolated 114 bacterial strains (8 moderately halophilic, 19 slightly halophilic, 87 halotolerant) from the samples on media (marine agar 2216, International Streptomyces Project medium 2 and 5, nutrient and humic acid agars) supplemented with 5% to 20% (w/v) NaCl. On the basis of morphological, physiological and biochemical characteristics, we selected 61 strains to perform a phylogenetic analysis based on 16S rRNA gene sequences. Results showed that 61 isolates represented 41 species, belonging to 18 genera (Actinomadura, Arthrobacter, Bacillus, Brachybacterium, Brevibacterium, Erwinia, Halobacillus, Jeotgalibacillus, Microbacterium, Microbulbifer, Nocardia, Pseudomonas, Rheinheimera, Rhodococcus, Sphingomonas, Staphylococcus, Streptomyces, Yaniella) of 16 families ( Alteromonadaceae, Bacillaceae, Brevibacteriaceae, Chromatiaceae, Dermabacteraceae, Enterobacteriaceae, Microbacteriaceae, Micrococcaceae, Nocardiaceae, Planococcaceae, Pseudomonadaceae, Sphingomonadaceae, Staphylococcaceae, Streptomycetaceae, Thermomonosporaceae, Yaniellaceae) in three phyla (Actinobacteria, Firmicutes, Proteobacteria). The most abundant and diverse isolates were within the phylum Firmicutes (38 strains; 62.3%) and the phylum Actinobacteria (18 strains; 29.5%). The phylogenetic distance matrix results suggested that there were obvious genetic divergences between most isolates and their closestly related type strains (16S rRNA gene sequence similarities ranged from 96.9% to 99.8%), and that, out of 61 isolates, at least 7 strains (JSM 070026, JSM 081004, JSM 081006, JSM 081008, JSM 083058, JSM 083085, JSM 084035) should represent 7 potential novel species within 6 characterized genera (Yaniella, Bacillus, Jeotgalibacillus, Sphingomonas, Rheinheimera, Microbulbifer). CONCLUSION: The results presented here showed that there is abundant diversity of halophilic and halotolerant bacteria, as well as a number of novel species in non-saline soil collected from Xiaoxi National Natural Reserve, Hunan Province, China.

3-(2-Bromo-eth-oxy)-4-(4-bromo-phen-yl)furan-5(2H)-one.

In the title compound, C(12)H(10)Br(2)O(3), the dihedral angle between the furan-5(2H)-one ring and the benzene ring is 1.2 (3)°. Two intra-molecular C-H⋯O inter-actions occur in the mol-ecule, both of which generate S(6) rings. The bromo-ethyl fragment is disordered over two sets of sites in a 0.773 (8):0.227 (8) ratio. In the crystal, inversion dimers linked by pairs of C-H⋯π inter-actions occur.

Halobacillus salsuginis sp. nov., a moderately halophilic bacterium from a subterranean brine.

A moderately halophilic, endospore-forming, Gram-positive, catalase- and oxidase-positive, motile, rod-shaped, aerobic bacterium, designated strain JSM 078133(T), was isolated from a subterranean brine sample collected from a salt mine in Hunan Province, China. Strain JSM 078133(T) was able to grow with 1-23 % (w/v) total salts (optimum, 5-10 %) and at pH 6.0-10.0 (optimum, pH 7.5) and 10-45 degrees C (optimum, 30 degrees C). meso-Diaminopimelic acid was present in the cell-wall peptidoglycan. The predominant respiratory quinone was menaquinone 7 (MK-7) and the major cellular fatty acids were anteiso-C(15 : 0), anteiso-C(17 : 0) and iso-C(15 : 0). The genomic DNA G+C content of strain JSM 078133(T) was 42.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain JSM 078133(T) belongs to the genus Halobacillus, and was related most closely to the type strains of Halobacillus campisalis (97.1 % sequence similarity), Halobacillus alkaliphilus (97.0 %) and Halobacillus yeomjeoni (96.8 %). The combination of phylogenetic analysis, DNA-DNA relatedness data, phenotypic characteristics and chemotaxonomic data supported the view that strain JSM 078133(T) represents a novel species of the genus Halobacillus, for which the name Halobacillus salsuginis sp. nov. is proposed. The type strain is JSM 078133(T) (=DSM 21185(T)=KCTC 13236(T)).

Halobacillus hunanensis sp. nov., a moderately halophilic bacterium isolated from a subterranean brine.

A moderately halophilic, Gram-positive, catalase- and oxidase-positive, rod-shaped, aerobic bacterium, designated strain JSM 071077(T), was isolated from a subterranean brine sample collected from a salt mine in Hunan Province, China. Cells were motile by means of peritrichous flagella and formed ellipsoidal endospores lying in subterminal swollen sporangia. Strain JSM 071077(T) was able to grow with 2-25% (w/v) total salts (optimum, 5-10%), at pH 6.0-10.0 (optimum, pH 7.5) and 10-40 degrees C (optimum, 25-30 degrees C). meso-Diaminopimelic acid was present in the cell-wall peptidoglycan. The predominant menaquinone was MK-7, and the major cellular fatty acids were anteiso-C(15:0), anteiso-C(17:0) and iso-C(15:0). The genomic DNA G+C content was 41.8 mol%. Phylogenetic analyses based on 16S rRNA gene sequence comparisons revealed that strain JSM 071077(T) should be assigned to the genus Halobacillus, being related most closely to the type strain of Halobacillus naozhouensis (98.8% sequence similarity), and the two strains formed a distinct subline in the neighbor-joining, minimum-evolution and maximum-parsimony phylogenetic trees. The sequence similarities between the novel isolate and the type strains of other recognized Halobacillus species ranged from 97.6% (with Halobacillus alkaliphilus) to 95.2% (with Halobacillus kuroshimensis). The results of the phylogenetic analyses, combined with DNA-DNA relatedness data, phenotypic characteristics and chemotaxonomic information, support that strain JSM 071077(T) represents a new species of the genus Halobacillus, for which the name Halobacillus hunanensis sp. nov. is proposed. The type strain is JSM 071077(T) (=DSM 21184(T) = KCTC 13235(T)).

Virgibacillus sediminis sp. nov., a moderately halophilic bacterium isolated from a salt lake in China.

A Gram-positive, moderately halophilic, alkalitolerant, strictly aerobic, oxidase- and catalase-positive, rod-shaped bacterium, strain YIM kkny3T, was isolated from a sediment sample collected from a salt lake in the Qaidam Basin of north-west China. Cells were motile by means of peritrichous flagella and formed ellipsoidal endospores lying in subterminal swollen sporangia. Growth occurred with 1-20% (w/v) total salts (optimum, 5-10%) and at pH 6.0-10.5 (optimum, pH 7.5-8.0) and 10-55 degrees C (optimum, 35-40 degrees C). It was unable to grow with NaCl as the only salt. meso-Diaminopimelic acid was present in the cell-wall peptidoglycan. The strain contained menaquinone 7 (MK-7) as the predominant respiratory quinone and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid as polar lipids. The major cellular fatty acids were anteiso-C15:0 and anteiso-C17:0. The DNA G+C content was 40.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM kkny3T belonged to the genus Virgibacillus, and was most closely related to the type strains of Virgibacillus olivae (97.1% similarity), Virgibacillus marismortui (97.0%) and Virgibacillus kekensis (96.8%). Levels of DNA-DNA relatedness between strain YIM kkny3T and the type strains of V. olivae, V. marismortui and V. kekensis were 12.4, 10.6 and 15.7%, respectively. The combination of phylogenetic analysis, genotypic data, phenotypic characteristics and chemotaxonomic differences indicated that strain YIM kkny3T represents a novel species of the genus Virgibacillus, for which the name Virgibacillus sediminis sp. nov. is proposed. The type strain is YIM kkny3T (=CCTCC AA 207023T=DSM 19797T=KCTC 13193T).

Saccharospirillum salsuginis sp. nov., a gammaproteobacterium from a subterranean brine.

A novel Gram-negative, non-sporulating, moderately halophilic, facultatively alkaliphilic, catalase- and oxidase-positive, obligately aerobic bacterium, strain YIM-Y25(T), was isolated from a subterranean brine sample collected from a salt mine in Yunnan, south-west China. Cells were spirilla, motile by monopolar flagella, with meso-diaminopimelic acid in the cell-wall peptidoglycan. Growth occurred with 1-15 % (w/v) NaCl (optimum 5 %), at pH 6.0-10.0 (optimum pH 8.0) and at 15-50 degrees C (optimum 35-40 degrees C). Ubiquinone Q-8 was the predominant respiratory quinone. Polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylamine and an unidentified phospholipid. The major cellular fatty acids were C(18 : 1)omega7c, iso-C(16 : 0), C(16 : 0) and C(16 : 1). The genomic DNA G+C content was 58.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM-Y25(T) was most closely related to the type strain of the sole recognized species of the genus Saccharospirillum, Saccharospirillum impatiens EL-105(T) (sequence similarity 97.0 %), and these two strains formed a robust lineage in the phylogenetic tree. The level of DNA-DNA relatedness between them was 12.6 %. The combination of phylogenetic analysis, phenotypic differences, chemotaxonomic properties and DNA-DNA hybridization data supported the view that this strain represents a novel species of the genus Saccharospirillum, for which the name Saccharospirillum salsuginis sp. nov. is proposed, with YIM-Y25(T) (=CCTCC AA 207033(T) =KCTC 22169(T)) as the type strain.