Vitamin K Insufficiency in the Indian Population: Pilot Observational Epidemiology Study.

BACKGROUND: The fat-soluble K vitamins K1 and K2 play an essential role in the blood coagulation cascade and are made available predominantly through selective dietary intakes. They are less known for their nonessential roles in a family of vitamin K-dependent proteins that promote various functions of organs and systems in the body. A lack of vitamin K can characterize vitamin and nutritional element insufficiency, which is different from a clinically apparent vitamin deficiency. OBJECTIVE: This epidemiological study evaluated the nutritional status of vitamin K in a sample of the Indian population and vitamin K content in staple Indian foods. METHODS: Serum levels of vitamin K1 and vitamin K2 in the form of menaquinone-7 (MK-7) were assessed via high-performance liquid chromatography coupled with fluorescence detection in 209 patients with type 2 diabetes, 50 healthy volunteers, and common staple foods in India. RESULTS: After comparing populations with high and low serum vitamin K levels from various geographical regions, our results indicated that the sample of healthy Indian individuals and the sample of Indian patients with type 2 diabetes had low (insufficient) levels of vitamin K2 (MK-7; range 0.3-0.4 ng/mL). No significant differences existed in vitamin K1-related and MK-7-related values between healthy male and female subjects, between male and female subjects with diabetes, and between the healthy sample and the sample of patients with diabetes. The staple, commonly consumed Indian foods that were tested in this study had undetectable levels of vitamin K2, while levels of vitamin K1 varied widely (range 0-37 µg/100 g). CONCLUSIONS: Based on our sample's low serum levels of vitamin K2 (MK-7) as well as the low levels of vitamin K2 in their typical diet, we propose that the general Indian population could benefit from the consumption of vitamin K2 in the form of MK-7 supplements. TRIAL REGISTRATION: Clinical Trials Registry - India CTRI/2019/05/014246; http://ctri.nic.in/Clinicaltrials/showallp.php?mid1=21660&EncHid=&userName=014246; Clinical Trials Registry - India CTRI/2019/03/018278; http://ctri.nic.in/Clinicaltrials/showallp.php?mid1=32349&EncHid=&userName=018278.

Simultaneous Determination of Vitamins D3 (Calcitriol, Cholecalciferol) and K2 (Menaquinone-4 and Menaquinone-7) in Dietary Supplements by UHPLC.

The content and composition of dietary supplements is of great interest due to their increasing consumption and variety of available brand offered in the market. Accurate determination of vitamins is important for the improvement of dietary supplement quality and nutrition assessments. In this regard, the simultaneous determination of vitamin D3 (calcitriol-CT and cholecalciferol-CHL) and K2 (menaquinone-4-MK-4 and menaquinone-7-MK-7) in dietary supplements was developed by using ultra-high-pressure liquid chromatography (UHPLC). The overall runtime per sample was above 35 min, with the retention times of 2.40, 6.59, 7.06, and 32.6 min for vitamin D3 (CT and CHL) and vitamin K2 (MK-4 and MK-7), respectively. The limits of detection and limits of quantification for the target nutritional compounds ranged between 0.04-0.05 µg/mL, respectively. The validation results indicated that the method had reasonable linearity (R2 ≥ 0.9990), good recovery (>82%), satisfactory intra-day precision (≤1.9%) and inter-day precision (≤3.5%), and high selectivity and specificity. The validated UHPLC method was demonstrated to be precise, accurate, and robust for the simultaneous determination of vitamins D3 (CT and CHL) and K2 (MK-4 and MK-7) in dietary supplements.

Planococcus lenghuensis sp. nov., an oil-degrading bacterium isolated from petroleum-contaminated soil.

A Gram-staining-positive and aerobic coccus with the ability to degrade petroleum bacterium, designated Y42T, was isolated from the Lenghu oil field located in the northern margin of the Qaidam Basin. Phylogenetic and signature nucleotides analyses revealed that strain Y42T belongs to the genus Planococcus. The multiple sequence alignments of 16S rRNA and housekeeping genes showed that strain Y42T formed a distinct lineage with the other Planococcus clade. The average nucleotide identity (ANI) and DNA-DNA hybridization values (DDH) between strain Y42T and the reference strains were 69.5-70.1 and 19.4-21.7%, respectively, which values were below the threshold for species delineation. The major fatty acids of strain Y42T were anteiso-C15:0. The respiratory quinone was MK-7 (71.8%) as the predominant menaquinone followed the MK-6 (28.2%) and the cell-wall hydrolysates contained LL-diaminopimelic acid. The polar lipid was composed of diphosphatidyl glycerol, phosphatidyl glycerol, phosphoglycolipid, aminophospholipid and four unidentified lipids. The peptidoglycan type was A4α (L-Lys-D-Glu). The strain Y42T possessed larger genome (approximately 4 MB) and revealed obvious differences for the abundance of the COG categories compared with the other Planococcus bacteria. Also, the strain Y42T also possessed more unique orthologous proteins. The structural characteristics of the strain Y42T genome provided a competitive advantage for better survival in petroleum-polluted environments. Combined with the 16S rRNA gene and genome sequence, phenotypic as well as chemotaxonomic characterisations, strain Y42T is considered to represent a novel species of the genus Planococcus, for which the name Planococcus lenghuensis sp. nov. be proposed. The type strain is Y42T (= CGMCC 1.15921T = JCM 32719T).

Cis and trans isomers of the vitamin menaquinone-7: which one is biologically significant?

Recently, several studies have indicated that an adequate intake of menaquinone-7 (MK-7) offers numerous health benefits. However, the low availability of MK-7 in the diet necessitates the development of dietary supplements or functional food products to complement natural food sources and meet the daily intake requirements. Like most biological molecules, MK-7 can exist as geometric isomers that can occur in the cis, trans, and cis/trans forms; however, only the all-trans form is biologically significant. MK-7 is traditionally produced through bacterial fermentation, but various synthetic preparations have lately become available. The isomer composition in the final product is influenced by numerous factors, including the methods of production and purification, as well as particular environmental and storage conditions. The MK-7 profile obtained from the various production methods has not yet been elucidated, and the ideal method for the synthesis of the all-trans form of the vitamin is also debatable. Consequently, the quantification of the MK-7 profile of various products is necessary to develop an understanding of the factors that influence the proportion of isomers that are obtained in different preparations. Several possible methods exist for the quantification of MK-7 isomers, and of these, liquid chromatography in conjunction with mass spectrometry techniques appears to be the most promising. Evaluation of the isomer composition is an important consideration, as only the all-trans form sustains biological activity. Furthermore, knowledge of the prominent factors that influence the MK-7 composition may also enable their manipulation to obtain a more favorable MK-7 profile in the final product.

Evaluation of vitamin K status and rationale for vitamin K supplementation in dialysis patients.

The cardinal biological role of vitamin K is to act as cofactor for the carboxylation of a number of vitamin K-dependent proteins, some of which are essential for coagulation, bone formation and prevention of vascular calcification. Functional vitamin K deficiency is common and severe among dialysis patients and has garnered attention as a modifiable risk factor in this population. However, no single biochemical parameter can adequately assess vitamin K status. For each biological function of vitamin K, the degree of carboxylation of the relevant vitamin K-dependent protein most accurately reflects vitamin K status. Dephosphorylated uncarboxylated matrix Gla protein (dp-ucMGP) is the best biomarker for vascular vitamin K status when cardiovascular endpoints are studied. Dp-ucMGP levels are severely elevated in haemodialysis patients and correlate with markers of vascular calcification and mortality in some but not all studies. The aetiology of vitamin K deficiency in haemodialysis is multifactorial, including deficient intake, uraemic inhibition of the vitamin K cycle and possibly interference of vitamin K absorption by phosphate binders. The optimal vitamin K species, dose and duration of supplementation to correct vitamin K status in dialysis patients are unknown. Dp-ucMGP levels dose-proportionally decrease with supraphysiological vitamin K2 supplementation, but do not normalize even with the highest doses. In the general population, long-term vitamin K1 or K2 supplementation has beneficial effects on cardiovascular disease, bone density and fracture risk, and insulin resistance, although some studies reported negative results. In haemodialysis patients, several trials on the effects of vitamin K on surrogate markers of vascular calcification are currently ongoing.

Menaquinone 7 Stability of Formulations and Its Relationship with Purity Profile.

Menaquinone-7 (MK7) is a member of the vitamin K family in which interest has considerably increased over the last decade, mainly due to its beneficial role in human health. MK7 can be produced by synthesis or fermentation, and its purity profile can differ depending on methodologies and extraction procedures. Finished formulations show a high heterogeneity of purity profiles, as well as frequent discrepancies in the nominal content, compared to the actual title. The present study compared purity profiles of different raw material and related them to their stability in normal (12 months/25 °C/60%RH) and accelerated conditions (6 months/40 °C/75% RH) in order to test their performance in the presence of different common excipients. Results showed higher purity profile results in enhanced stability, and this could explain title discrepancies found in finished products, which are present on the market worldwide.

Analysis of Menaquinone-7 Content and Impurities in Oil and Non-Oil Dietary Supplements.

Rapid, global technological development has caused the food industry to use concentrated food component sources like dietary supplements ever more commonly as part of the human diet. This study analysed the menaquinone-7 (MK-7) content of dietary supplements in oil capsule and hard tablet forms. A novel method for separating and measuring geometric isomers of MK-7 in dietary supplements was developed and validated. Eleven different isomers of cis/trans- menaquinone-7 were identified. Identification of cis/trans isomers was performed by combination of HPLC, UPLC and HRMS-QTOF detection, whereas their quantities were determined by DAD detection. The content of menaquinone impurities was ascertained, including cis/trans- menaquinone-6 isomers (5.5⁻16.9 µg per tablet/capsule) and cis/trans-menaquinone-7 isomers (70.9⁻218.7 µg tablet/capsule), which were most likely formed during the chemical synthesis of the menaquinone-7. The all-trans MK-7 content was lower than the isomeric form and often lower than what the labels declared. A new method of quantification, developed and validated for menaquinones in oil capsules, provided on average 90% recovery and a limit of quantification (LOQ) of approximately 1 µg mL−1.

Flavobacterium panacisoli sp. nov., isolated from soil of a ginseng field.

A novel bacterial strain, designated DCY70(T), was isolated from soil of a ginseng field in Republic of Korea and was characterized in order to determine its taxonomic position. The strain was Gram-reaction negative, yellow-pigmented, rod-shaped and catalase- and oxidase-positive. Based on the 16S rRNA gene sequence similarity, strain DCY70(T) was shown to belong to the genus Flavobacterium, most closely related to Flavobacterium oncorhynchi 631-08(T) (98.4 %), Flavobacterium plurextorum 1126-1H-08(T) (97.9 %), Flavobacterium chilense LM-09-Fp(T) (97.9 %) and Flavobacterium chungangense CJ(T) (97.7 %). The chemotaxonomic characteristics showed only menaquinone-6 (MK-6), iso-C15:0, iso-C15:0 3OH, iso-C17:0 3OH and summed feature 3 as major cellular fatty acids. Polar lipids were phosphatidylethanolamine (PE), two unidentified aminolipids, four unidentified polar lipids and one unidentified phospholipid. The DNA G+C content was 34.9 mol%. Based on the phylogenetic, phenotypic and genotypic data, a novel species, Flavobacterium panacisoli sp. nov., is proposed (=KCTC 32393(T) = JCM 19162(T)).

Paenibacillus enshidis sp. nov., Isolated from the Nodules of Robinia pseudoacacia L.

A Gram-positive, motile, endospore-forming, rod-shaped bacterium, designated RP-207(T), was isolated from the nodules of Robinia pseudoacacia L. plants planted in Enshi District, Hubei, PR China. Phylogenetic analyses based on the 16S rRNA gene sequence showed that the novel strain was affiliated to the genus Paenibacillus, with its closest relatives being Paenibacillus xylanilyticus XIL14(T) (95.6%), Paenibacillus peoriae DSM8320(T) (95.3%) and Paenibacillus polymyxa DSM 36(T) (95.3%). The DNA G+C content was 47.0 mol%. DNA-DNA hybridization value between strain RP-207(T) and P. xylanilyticus XIL14(T) was 40.1%. The diamino acid found in the cell wall peptidoglycan was meso-diaminopimelic. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, an unidentified amino-phospholipid and an unknown phospholipid. The predominant menaquinone was menaquinone-7 (MK-7), and the major fatty acid was anteiso-C15:0 and C16:0. On the basis of its physiological and biochemical characteristics and the level of DNA-DNA hybridization, strain RP-207(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus enshidis sp. nov. is proposed. The type strain is RP-207(T) (=CCTCC AB 2013275(T) = KCTC 33519(T)).

Nocardia rhizosphaerae sp. nov., a novel actinomycete isolated from the coastal rhizosphere of Artemisia Linn., China.

A novel actinomycete, designated strain KLBMP S0043(T), was isolated from the rhizosphere soil of Artemisia Linn. collected from the coastal region of Lianyungang, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP S0043(T) is a member of the genus Nocardia. The 16S rRNA gene sequence similarity indicated that strain KLBMP S0043(T) is closely related to Nocardia asteroides NBRC 15531(T) (97.61 %) and Nocardia neocaledoniensis SBHR OA6(T) (97.38 %); similarity to other type strains of the genus Nocardia was found to be less than 97.2 %. The organism has chemical and morphological features consistent with its classification in the genus Nocardia such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was identified as MK-8(H4ω-cycl). Mycolic acids were detected. The diagnostic phospholipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The predominant cellular fatty acids were identified as C16:0, C18:0, C18:1ω9c, 10-methyl C18:0 [tuberculostearic acid (TBSA)] and summed feature 3 (C16:1ω7c/C16:1ω6c). The G+C content of the genomic DNA was determined to be 71.4 mol%. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the strain from its most closely related strains. Based on morphological, chemotaxonomic and phylogenetic data, strain KLBMP S0043(T) is considered to represent a novel species of the genus Nocardia, for which the name Nocardia rhizosphaerae sp. nov. is proposed. The type strain is KLBMP S0043(T) (=CGMCC 4.7204 (T) = KCTC 29678(T)).

Acción de la vitamina K sobre la salud ósea / Action of vitamin K on bone health

Objetivos: Se admite hoy en día que la vitamina K tiene un papel importante en la salud ósea. Es necesaria para la gamma-carboxilación de la osteocalcina (la proteína no colágena más importante en el hueso), consiguiendo que la osteocalcina funcione. Hay dos formas importantes de la vitamina K (vitamina K1 y vitamina K2), que provienen de diferentes fuentes y tienen diferentes actividades biológicas. Estudios epidemiológicos sugieren que una dieta con niveles altos de vitamina K se asocia con un menor riesgo de fracturas de cadera en hombres ancianos y mujeres. Sin embargo, ensayos clínicos controlados y aleatorizados, realizados con suplementos de vitamina K1 o K2 en la población blanca, no muestran un aumento en la densidad mineral ósea (DMO) en la mayoría de las diferentes partes del esqueleto. Los suplementos con vitamina K1 y K2 pueden reducir el riesgo de fractura, pero los ensayos clínicos que incluyen las fracturas como resultado final tienen limitaciones metodológicas, por lo que se necesitarían ensayos clínicos con mayor número de pacientes y mejor diseñados para comprobar la eficacia de la vitamina K1 y K2 en las fracturas. Como conclusión, podríamos decir que actualmente no existe una evidencia suficiente para recomendar el uso rutinario de suplementos de vitamina K para la prevención de la osteoporosis y las fracturas en mujeres postmenopáusicas (AU)

Objetives: Nowadays it is recognised that vitamin K plays an important role in bone health. It is necessary for the gamma-carboxylation of osteocalcin (the most important non-collagen protein in the bone), making the osteocalcin function. There are two important forms of vitamin K (vitamin K1 and vitamin K2), which come from different sources and have different biological activity. Epidemiological studies suggest that a diet with high levels of vitamin K is associated with a lower risk of hip fractures in older men and in women. However, controlled randomised clinical trials, carried out with supplements of vitamin K1 or K2 in the white population do not show an increase in bone mineral density (BMD) in most of the different areas of the skeleton. Supplementation with vitamin K1 and K2 may reduce the risk of fracture, but the clinical trials which include fractures as a final result have methodological limitations, so clinical trials with greater numbers of patients, and which are better designed, would be needed in order to prove the efficacy of vitamin K1 and K2 in relation to fractures. In conclusion, we may say that there is currently insufficient evidence to recommend the routine use of vitamin K for the prevention of osteoporosis and fractures in postmenopausal women (AU)

Sphaerisporangium dianthi sp. nov., an endophytic actinomycete isolated from a root of Dianthus chinensis L.

A novel actinomycete, designated strain NEAU-CY18(T), was isolated from the root of a Chinese medicinal plant Dianthus chinensis L and subjected to a polyphasic taxonomic study. The novel strain was found to develop spherical sporangia with non-motile spores on aerial mycelium. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The whole-cell sugars were identified as madurose, mannose, ribose, galactose and glucose. The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, hydroxy-phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and an unidentified phospholipid. The predominant menaquinones were identified as MK-9(H4), MK-9(H2) and MK-9(H6). The major fatty acids were identified as C17:0 10-methyl, iso-C16:0 and C16:0. EzTaxon-e analysis of the 16S rRNA gene sequence indicated that the strain belongs to the genus Sphaerisporangium and was most closely related to Sphaerisporangium cinnabarinum JCM 3291(T) (98.9 %) and Sphaerisporangium melleum JCM 13064(T) (98.3 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-CY18(T) forms a monophyletic clade with S. cinnabarinum JCM 3291(T), an association that was supported by a bootstrap value of 97 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. Comparisons of some phenotypic properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. cinnabarinum JCM 3291(T) and S. melleum JCM 13064(T). Therefore, it is concluded that strain NEAU-CY18(T) represents a novel Sphaerisporangium species, for which the name Sphaerisporangium dianthi sp. nov. is proposed. The type strain is NEAU-CY18(T) ( = CGMCC 4.7132(T) = DSM 46736(T)).

Paenibacillus yonginensis sp. nov., a potential plant growth promoting bacterium isolated from humus soil of Yongin forest.

Strain DCY84(T), a Gram-stain positive, rod-shaped, aerobic, spore-forming bacterium, motile by means of peritrichous flagella, was isolated from humus soil from Yongin forest in Gyeonggi province, South Korea. Strain DCY84(T) shared the highest sequence similarity with Paenibacillus barengoltzii KACC 15270(T) (96.86 %), followed by Paenibacillus timonensis KACC 11491(T) (96.49 %) and Paenibacillus phoenicis NBRC 106274(T) (95.77 %). Strain DCY84(T) was found to able to grow best in TSA at temperature 30 °C, at pH 8 and at 0.5 % NaCl. MK-7 menaquinone was identified as the isoprenoid quinone. The major polar lipids were identified as phosphatidylethanolamine, an unidentified aminophospholipid, two unidentified aminolipids and an unidentified polar lipid. The peptidoglycan was found to contain the amino acids meso-diaminopimelic acid, alanine and D-glutamic acid. The major fatty acids of strain DCY84(T) were identified as branched chain anteiso-C15:0, saturated C16:0 and branched chain anteiso-C17:0. The cell wall sugars of strain DCY84(T) were found to comprise of ribose, galactose and xylose. The major polyamine was identified as spermidine. The DNA G+C content was determined to be 62.6 mol%. After 6 days of incubation, strain DCY84(T) produced 52.96 ± 1.85 and 72.83 ± 2.86 µg/ml L-indole-3-acetic acid, using media without L-tryptophan and supplemented with L-tryptophan, respectively. Strain DCY84(T) was also found to be able to solubilize phosphate and produce siderophores. On the basis of the phenotypic characteristics, genotypic analysis and chemotaxonomic characteristics, strain DCY84(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus yonginensis sp. nov. is proposed. The type strain is DCY84(T) (=KCTC 33428(T) = JCM 19885(T)).

Streptomyces kebangsaanensis sp. nov., an endophytic actinomycete isolated from an ethnomedicinal plant, which produces phenazine-1-carboxylic acid.

A spore-forming streptomycete designated strain SUK12(T) was isolated from a Malaysian ethnomedicinal plant. Its taxonomic position, established using a polyphasic approach, indicates that it is a novel species of the genus Streptomyces. Morphological and chemical characteristics of the strain were consistent with those of members of the genus Streptomyces. Analysis of the almost complete 16S rRNA gene sequence placed strain SUK12(T) in the genus Streptomyces where it formed a distinct phyletic line with recognized species of this genus. The strain exhibited highest sequence similarity to Streptomyces corchorusii DSM 40340(T) (98.2 %) followed by Streptomyces chrestomyceticus NRRL B-3310(T) (98.1 %). The G+C content of the genomic DNA was 74 mol%. Chemotaxonomic data [MK-9(H8) as the major menaquinone; LL-diaminopimelic acid as a component of cell-wall peptidoglycan; C12 : 0, C14 : 0, C15 : 0 and C17 : 1 as the major fatty acids; phospholipid type II] supported the affiliation of strain SUK12(T) to the genus Streptomyces. The results of the phylogenetic analysis and phenotypic data derived from this and previous studies allowed the genotypic and phenotypic differentiation of strain SUK12(T) from the related species of the genus Streptomyces. The DNA-DNA relatedness value between strain SUK12(T) and S. corchorusii DSM 40340(T) is 18.85±4.55 %. Strain SUK12(T) produces phenazine-1-carboxylic acid, known as tubermycin B, an antibacterial agent. It is proposed, therefore, that strain SUK12(T) ( = DSM 42048(T) = NRRL B-24860(T)) be classified in the genus Streptomyces as the type strain of Streptomyces kebangsaanensis sp. nov.

Chryseobacterium yeoncheonense sp. nov., with ginsenoside converting activity isolated from soil of a ginseng field.

A Gram-staining negative, aerobic, non-motile, non-flagellate, yellow-pigmented, rod-shaped bacterial strain, designated strain DCY67(T), was isolated from ginseng field in Republic of Korea. Strain DCY67(T) contained ß-glucosidase activity which converts ginsenoside Rb1 to compound K. Optimum growth of DCY67(T) occurred at 30 °C and pH 6.0-6.5. Analysis of the 16S rRNA gene sequences revealed that strain DCY67(T) belonged to the family Flavobacteriaceae and was most closely related to Chryseobacterium ginsenosidimutans THG 15(T) (97.5 %). The genomic DNA G+C content was 36.1 mol%. The predominant quinones were MK-6 (90.9 %) and MK-7 (9.15 %). The major fatty acids were iso-C15:0, summed feature 3 (containing C16:1 ω7c and/or C16:1 ω6c) and iso-C17:0 3-OH. On the basis of these phenotypic, genotypic and chemotaxonomic studies, strain DCY67(T) represents a novel species of the genus Chryseobacterium, for which, name Chryseobacterium yeoncheonense sp. nov. proposed the type strain is DCY67(T) (=KCTC 32090(T) = JCM 18516(T)).

Nocardioides panaciterrulae sp. nov., isolated from soil of a ginseng field, with ginsenoside converting activity.

A Gram-positive, coccoid to rod-shaped, non-spore-forming bacterium, designated Gsoil 958(T), was isolated from soil of a ginseng field located in Pocheon province in South Korea. This bacterium was characterized in order to determine its taxonomic position by using a polyphasic approach. Strain Gsoil 958(T) was observed to grow well at 25-30 °C and at pH 7.0 on R2A and nutrient agar without NaCl supplementation. Strain Gsoil 958(T) was determined to have ß-glucosidase activity and the ability to transform ginsenoside Rb1 (one of the dominant active components of ginseng) to F2 via gypenoside XVII and Rd. On the basis of 16S rRNA gene sequence similarity, strain Gsoil 958(T) was shown to belong to the family Nocardioidaceae and related most closely to Nocardioides koreensis MSL-09(T) (97.6 % 16S rRNA gene sequence similarity), Nocardioides aquiterrae GW-9(T) (97.0 %), and Nocardioides sediminis MSL-01(T) (97.0 %). The sequence similarities with other validly named species within the genus Nocardioides were less than 96.8 %. Strain Gsoil 958(T) was characterized chemotaxonomically as having LL-2,6-diaminopimelic acid in the cell-wall peptidoglycan, MK-8(H4) as the predominant menaquinone, and iso-C16:0, iso-C16:1 H, iso-C14:0, iso-C15:0 were identified as the major fatty acids. The G + C content of genomic DNA was determined to be 70.8 mol %. The chemotaxonomic properties and phenotypic characteristics supported the affiliation of strain Gsoil 958(T) to the genus Nocardioides. The results of both physiological and biochemical tests allowed for differentiation of strain Gsoil 958(T) from the recognized Nocardioides species. Therefore, strain Gsoil 958(T) is considered to represent a novel species of the genus Nocardioides, for which the name Nocardioides panaciterrulae sp. nov. is proposed, with the type strain Gsoil 958(T) (KACC 14271(T) = KCTC 19471(T) = DSM 21350(T)).

Niabella hirudinis and Niabella drilacis sp. nov., isolated from the medicinal leech Hirudo verbana.

Two Gram-negative, rod-shaped bacteria, strains E96(T) and E90(T), were isolated from medicinal leeches (Hirudo verbana) and characterized by a polyphasic taxonomic approach. Phylogenetic analysis based on the nearly full-length 16S rRNA gene sequence showed that the two strains shared 98.1% sequence similarity and were affiliated with the genus Niabella within the phylum Bacteroidetes, with 94.4-97.6% sequence similarity to type strains of species of the genus Niabella and highest sequence similarity to the type strain of Niabella aurantiaca (97.3 and 97.6%, respectively). Niabella-related 16S rRNA gene sequences were recently detected in the bladders of Hirudo verbana; however, no cultured representatives were so far available. Genomic fingerprint analysis using repetitive element primed (rep)- and randomly amplified polymorphic DNA (RAPD)-PCRs and DNA-DNA hybridization experiments clearly showed that the strains were different from each other (DNA-DNA relatedness values of 39.1%, reciprocal 28.0%) and from the type strains of N. aurantiaca (<19.7%) and Niabella tibetensis (<41.1%). Chemotaxonomic analyses confirmed the affiliation to the genus Niabella. Both strains contained MK-7 as the predominant menaquinone. The major fatty acids of both strains were iso-C15:0, iso-C15:1 G, iso-C17:0 3-OH and summed feature 3 (C16:1ω7c and/or iso-C15:0 2-OH), which is characteristic for the genus Niabella. Based on genotypic, chemotaxonomic and physiological characterization, we propose two novel species of the genus Niabella, Niabella hirudinis sp. nov., with strain E96(T) ( =DSM 25812(T) =CCM 8411(T) =LMG 26956(T)) as the type strain, and Niabella drilacis sp. nov., with strain E90(T) ( =DSM 25811(T) =CCM 8410(T) =LMG 26954(T)) as the type strain.

Chryseobacterium ginsengisoli sp. nov., isolated from the rhizosphere of ginseng and emended description of Chryseobacterium gleum.

Strain DCY 63(T), isolated from the rhizosphere of ginseng, was a Gram-reaction-negative, strictly aerobic, non-motile, catalase- and oxidase-positive rod. 16S rRNA gene sequence analysis revealed that strain DCY 63(T) belonged to the genus Chryseobacterium and was closely related to Chryseobacterium indoltheticum LMG 4025(T) (98.1 %), Chryseobacterium ginsenosidimutans THG 15(T) (98.0 %) and Chryseobacterium scophthalmum LMG 13028(T) (97.1 %). The major polar lipids of strain DCY 63(T) were phosphatidylethanolamine (PE), four unidentified aminolipids and three unidentified polar lipids, and the major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and summed feature 9 (iso-C17 : 1ω9c and/or C16 : 0 10-methyl). The predominant isoprenoid quinone of strain DCY 63(T) was MK-6 and the G+C content of the genomic DNA was 31.6 mol%. The DNA-DNA relatedness values between strain DCY 63(T) and C. indoltheticum LMG 4025(T), C. ginsenosidimutans THG 15(T) and C. scophthalmum LMG 13028(T) were less than 30 %. On the basis of these data, strain DCY 63(T) is considered to represent a novel species of the genus Chryseobacterium, for which the name Chryseobacterium ginsengisoli sp. nov. is proposed. The type strain is DCY 63(T) (= KCTC 23760(T) = JCM 18019(T)). An emended description of Chryseobacterium gleum is also proposed.

Mucilaginibacter herbaticus sp. nov., isolated from the rhizosphere of the medicinal plant Angelica sinensis.

A strictly aerobic, Gram-staining-negative, non-motile and rod-shaped bacterial strain, DR-9(T), was isolated from rhizosphere soil of the medicinal herb Angelica sinensis. Strain DR-9(T) grew at 20-40 °C, at pH 4.0-9.0 and in the presence of 0-1 % (w/v) NaCl. The major fatty acids were iso-C15 : 0 and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c and/or iso-C15 : 0 2-OH), MK-7 was the major isoprenoid quinone, and phosphatidylethanolamine and an unidentified aminophospholipid were the major polar lipids. A phylogenetic tree based on 16S rRNA gene sequences showed that strain DR-9(T) formed a lineage within the genus Mucilaginibacter and was closely related to Mucilaginibacter polysacchareus DRP28(T) (96.1 % sequence similarity), Mucilaginibacter myungsuensis HMD1056(T) (95.9 % sequence similarity), Mucilaginibacter ximonensis XM-003(T) (95.8 %) and Mucilaginibacter boryungensis BDR-9(T) (95.1 %). The status of strain DR-9(T) as a representative of a separate species was confirmed by DNA hybridization, with 38.6, 36.3 and 29.9 % DNA-DNA relatedness with M. polysacchareus DRP28(T), M. ximonensis XM-003(T) and M. boryungensis BDR-9(T), respectively. The genomic DNA G+C content of strain DR-9(T) was 49.8 %. These data suggest that strain DR-9(T) should be considered as a representative of a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter herbaticus sp. nov. is proposed. The type strain is DR-9(T) ( = KACC 16469(T) = NBRC 108839(T)).

Allonocardiopsis opalescens gen. nov., sp. nov., a new member of the suborder Streptosporangineae, from the surface-sterilized fruit of a medicinal plant.

An endophytic actinomycete, designated strain I10A-01259(T), was isolated from a surface-sterilized fruit of Lonicera maackii (Rupr.) Maxim., a medicinal plant, which was collected from a suburb of Beijing, China. Whole-cell hydrolysates of the isolate contained galactose and meso-diaminopimelic acid. The predominant phospholipids were phosphatidylglycerol and diphosphatidylglycerol; the menaquinones consisted mainly of MK-9, MK-11 and MK-12, with a minor amount of MK-10. The major fatty acids were iso-C16 : 0, anteiso-C15 : 0 and iso-C15 : 0. Comparative analysis of 16S rRNA gene sequences indicated that strain I10A-01259(T) was most closely related to Nocardiopsis arabia S186(T) (93.2 % sequence similarity), Thermobifida halotolerans YIM 90462(T) (93.0 %) and other strains of genera within the families Nocardiopsaceae and Thermomonosporaceae. On the phylogenetic tree based on 16S rRNA gene sequences, strain I10A-01259(T) fell within the radius of the suborder Streptosporangineae, in which the strain formed a distinct lineage next to the genera of the families Nocardiopsaceae and Thermomonosporaceae. Based on the data from our polyphasic taxonomic study, a novel genus and species, Allonocardiopsis opalescens gen. nov., sp. nov., are proposed within the suborder Streptosporangineae. The type strain of Allonocardiopsis opalescens is strain I10A-01259(T) ( = CPCC 203428(T)  = DSM 45601(T)  = KCTC 19844(T)).