Methylobacterium crusticola sp. nov., isolated from biological soil crusts.

A pink-pigmented, Gram-negative, rod-shaped, obligate aerobic bacterial strain, MIMD6T, was isolated from biological soil crusts in PR China. Cells grew at 20-37 °C (optimum, 30 °C), at pH 6-8 (optimum, pH 7) and with 0-1 % (w/v) NaCl (optimum, 0 %). Strain MIMD6T could use methanol or formate as a sole carbon source to grow, and carried methanol dehydrogenase genes mxaF and xoxF, supporting its methylotrophic metabolism. The respiratory quinone was ubiquinone Q-10, the major fatty acids were C18 : 1ω7c (87.3 %), and the major polar lipids were diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, one unknown aminolipid and one unidentified glycolipid. The results of phylogenetic analyses based on the sequences of the 16S rRNA gene, seven housekeeping genes (dnaK, recA, rimO, rpIK, rpmG, rpsR and rpoB) and methanol dehydrogenase genes indicated that strain MIMD6T formed a phylogenetic linage with members of the genus Methylobacterium. Strain MIMD6T was most closely related to Methylobacterium isbiliense DSM 17168T and Methylobacterium nodulans LMG 21967T with 16S rRNA gene sequence similarities of 95.7 and 95.2 %, respectively. The genomic DNA G+C content calculated via draft genome sequencing was 73.0 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain MIMD6T and the type strains of other Methylobacterium species were 70.7-82.0 and 24.6-30.0 %, respectively. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain MIMD6T represents a novel species of the genus Methylobacterium, for which the name Methylobacterium crusticola sp. nov. is proposed. The type strain is MIMD6T (=KCTC 52305T=MCCC 1K01311T).

Aerophototrophica crusticola gen. nov., sp. nov., isolated from desert biocrusts.

A pink-pigmented, Gram-stain-negative, rod-shaped, strictly aerobic bacterial strain MIMtkB3T, was isolated from moss crusts in Hunshandake desert of China. Cells grew at 15-45 °C (optimum of 28 °C), at pH of 6.0-8.5 (optimum of 7.0) and with 0-1.0 % (w/v) NaCl (optimum of 0 %). The strain could biosynthesize the green-coloured pigment bacteriochlorophyll a (BChl a). The respiratory quinone was ubiquinone Q-10, while C18 : 1 ω7c and C18 : 1 2OH were the major fatty acids. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminophospholipid, one unidentified phospholipid, three unidentified glycolipid and one unidentified lipid were the major polar lipids. Strain MIMtkB3T was most closely related to Oleisolibacter albus NAU-10T, Niveispirillum fermenti CC-LY736T, and Rhodocista centenaria SW of the family Rhodospirillaceae with 16S rRNA gene similarities of 93.09, 92.02 and 91.73%, respectively. The genomic DNA G+C content calculated on complete genome sequencing was 69.3 mol%. The average nucleotide identity between strain MIMtkB3T and its closely related type strains in Rhodospirillaceae was below 77.96 % and digital DNA-DNA hybridization lower than 24.70 %. Full light utilization pathway of aerobic anoxygenic phototrophic bacteria was identified in the genome. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, strain MIMtkB3T represents a novel genus of the family Rhodospirillaceae, for which the name Aerophototrophica crusticola gen. nov., sp. nov. is proposed. The type strain is MIMtkB3T (=KCTC 42633T=MCCC 1K00570T).

Rhizobium arenae sp. nov., isolated from the sand of Desert Mu Us, China.

A Gram-strain-negative, rod-shaped, motile bacterium, designated MIM27T, was isolated from the sand of the Mu Us Desert, PR China. The strain could grow at 4-45 °C (optimum, 37 °C), at pH 6.6-9.0 (optimum, 8.0) and in the presence of 0-3 % (w/v) NaCl (optimum, 0 % in RNA liquid medium). The results of phylogenetic analysis of 16S rRNA gene sequences indicated that the strain represented a member of the genus Rhizobium, with the highest similarity (96.5 %) to Rhizobium pakistanense BN-19T. The results of analysis of the sequences of the nitrogen fixation gene nifH and three housekeeping genes, recA, atpD and glnII, also indicated that MIM27T was most closely related to the species of the genus Rhizobiumwith validly published names but the similarities were low (≤90.7 %). MIM27T did not form nodules on Pisum sativum, Vicia faba, Astragalus sinicus and Phaseolus vulgaris. The major respiratory quinone of MIM27T was Q-10. The genomic DNA G+C content was 59.8 mol%. Major fatty acids of MIM27T were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C18 : 1ω7c 11-methyl, C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and summed feature 2 (C12 : 0 aldehyde and/or unknown ECL 10.9525). On the basis of the physiological, chemotaxonomic and phenotypic data, MIM27T is suggested to represent a novel species of the genus Rhizobium, for which the name Rhizobium arenae sp. nov. is proposed. The type strain is MIM27T (=KCTC 52299T=MCCC 1K03215T).

Mongoliibacter ruber gen. nov., sp. nov., a haloalkalitolerant bacterium of the family Cyclobacteriaceae isolated from a haloalkaline lake.

A novel haloalkalitolerant, rod-shaped bacterium, designated strain YIM 4-4T, was isolated from the surface water of the Dugerno lake, a haloalkaline lake in Inner Mongolia. The taxonomy of strain YIM 4-4T was investigated by a polyphasic approach. Strain YIM 4-4T was Gram-stain-negative, strictly aerobic, non-motile and formed red colonies. Optimal growth conditions were 28 °C, pH 8.0-11.0 and 0.5-2 % NaCl. The major respiratory quinone was menaquinone-7 (MK-7). The polar lipid profile was composed predominantly of phosphatidylethanolamine, six unidentified polar lipids, one phospholipid and one aminolipid. The predominant cellular fatty acids (>5 %) were iso-C15 : 0, iso-C17 : 1I/anteiso-C17 : 1B, iso-C16 : 1G, iso-C17 : 0 3-OH, C16 : 1ω7c/C16 : 1ω6c and iso-C16 : 1. The genomic DNA G+C content was 43.0 mol%. 16S rRNA gene sequence analysis indicated that the members of the genera Cecembia, Fontibacter, Aquiflexum and Indibacter of the family Cyclobacteriaceae (phylum Bacteroidetes) were the most closely related, with 16S rRNA gene sequence similarities ranging from 93.6 to 94.2 %. Other members of the family Cyclobacteriaceae showed sequence similarities < 93.0 %. On the basis of phenotypic, chemotaxonomic and phylogenetic properties, strain YIM 4-4T represents a novel species of a new genus, for which the name Mongoliibacter ruber gen. nov., sp. nov. is proposed. The type strain is YIM 4-4T ( = CCTCC AB 2012966T = DSM 27929T).

Hymenobacter terrenus sp. nov., isolated from biological soil crusts.

A Gram-stain-negative, non-spore-forming, short rod-shaped, non-motile, light-pink bacterial strain, MIMtkLc17T, was isolated from biological soil crusts collected in Liangcheng, Inner Mongolia. Growth of strain MIMtkLc17T was observed at 2-35 °C and in the presence of 1% (w/v) NaCl. Phylogenetic analysis of 16S rRNA gene sequences showed that sequence similarities between strain MIMtkLc17T and the type strains of species of the genus Hymenobacter ranged from 89.93% to 96.49%. Strain MIMtkLc17T can secrete mass polysaccharide. The major fatty acids of strain MIMtkLc17T were iso-C15 : 0, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), C16 : 1ω5c and summed feature 4 (iso-C17 : 1 I/anteiso-C17 : 1 B). The sole respiratory quinone was menaquinone MK-7. The G+C content of the chromosomal DNA was 57.8 mol%. The results of phylogenetic, chemotaxonomic and phenotypic characterization indicated that strain MIMtkLc17T can be distinguished from all known species of the genus Hymenobacter and represents a novel species of this genus, for which the name Hymenobacter terrenus sp. nov. is proposed. The type strain is MIMtkLc17T ( = MCCC 1K00507T = KCTC 42636T).

Mongoliicoccus roseus gen. nov., sp. nov., an alkaliphilic bacterium isolated from a haloalkaline lake.

Two pink, non-motile, aerobic, alkaliphilic, halotolerant, Gram-negative cocci, designated MIM28(T) and MIM29, were isolated from the surface water of a haloalkaline lake on the Mongolia Plateau. The isolates grew optimally at 30-33 °C, at pH 8-9 and with 3-4 % (w/v) NaCl. The isolates were chemoheterotrophic and could assimilate carbohydrates, organic acids and amino acids. The major respiratory quinone was menaquinone MK-7. The major polar lipids were phosphatidylcholine and phosphatidylethanolamine. The predominant cellular fatty acids were iso-C(15 : 0) (13.8-17.5 %), anteiso-C(15 : 0) (10.5-11.2 %), iso-C(16 : 0) (9.9-13.0 %), C(16 : 0) (4.3-4.6 %), iso-C(17 : 0) (3.8-5.3 %), anteiso-C(17 : 0) (3.7-7.1 %), C(17 : 1)ω6c (4.6-6.4 %), iso-C(17 : 0) 3-OH (4.6-5.8 %), summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c; 4.0-6.4 %) and summed feature 9 (iso-C(17 : 1)ω9c and/or C(16 : 0) 10-methyl; 10.4-12.5 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates were most closely related to Litoribacter ruber YIM CH208(T) (93.6 % 16S rRNA gene sequence similarity), the genus Echinicola (90.4-92 %) and other members of the family Cyclobacteriaceae (87.8-90 %). The DNA G+C contents of strains MIM28(T) and MIM29 were 62.8 and 62.2 mol%. On the basis of morphology, physiology, fatty acid composition, phylogeny and 16S rRNA gene sequence analysis, the isolates are assigned to a novel species of a new genus, for which we propose the name Mongoliicoccus roseus gen. nov., sp. nov.; the type strain of the type species is MIM28(T) (= ACCC 05511(T) = KCTC 19808(T)).

Bovine spongiform encephalopathy associated insertion/deletion polymorphisms of the prion protein gene in the four beef cattle breeds from North China.

Two insertion/deletion (indel) polymorphisms of the prion protein gene (PRNP), a 23-bp indel in the putative promoter region and a 12-bp indel within intron I, are associated with the susceptibility to bovine spongiform encephalopathy (BSE) in cattle. In the present study, the polymorphism frequencies of the two indels in four main beef cattle breeds (Hereford, Simmental, Black Angus, and Mongolian) from North China were studied. The results showed that the frequencies of deletion genotypes and alleles of 23- and 12-bp indels were lower, whereas the frequencies of insertion genotypes and alleles of the two indels were higher in Mongolian cattle than in the other three cattle breeds. In Mongolian cattle, the 23-bp insertion / 12-bp insertion was the major haplotype, whereas in Hereford, Simmental, and Black Angus cattle, the 23-bp deletion / 12-bp deletion was the major haplotype. These results demonstrated that Mongolian cattle could be more resistant to BSE, compared with the other three cattle breeds, because of its relatively low frequencies of deletion genotypes and alleles of 23- and 12-bp indel polymorphisms. Thus, this race could be important for selective breeding to improve resistance against BSE in this area.

[Dose-effect relationship in treatment of chronic neck pain with the direct moxibustion of small moxa cone].

OBJECTIVE: To evaluate the effectiveness in treatment of chronic neck pain with the direct moxibustion of small moxa cone and explore the dose-effect relationship in treatment of chronic neck pain with different small moxa cones. METHODS: A total of 120 patients with chronic neck pain were randomized into a 3-moxa-cone group, a 5-moxa-cone group, a 7-moxa-cone group and a sham-moxibustion group, 30 cases in each one. Fengchi (GB 20), Tianzhu (BL 10), Jingbailao (EX-HN 15), Jianzhongshu (SI 15) and Jianjing (GB 21) were selected in each of the groups. The direct moxibustion with 3, 5 and 7 moxa cones as well as the sham-moxibustion therapy were provided successively in each of the above groups. In the sham-moxibustion group, the lower 1/3 section of moxa cone (about 1.5 mm in length) was soaked in wanhua oil before used in treatment. In each group, the treatment was given twice a week, for 10 treatments totally. Separately, before treatment, after treatment and in 1-month follow-up, the scores of the Northwick Park neck pain questionnaire (NPQ), the scores of McGill pain questionnaire (MPQ) and the local pressure pain threshold (PPT) were observed in each group. After treatment and in 1-month follow-up, the therapeutic effects were evaluated in each group. RESULTS: After treatment and in 1-month follow-up, NPQ scores and MPQ scores in the 3-moxa-cone group, the 5-moxa-cone group and the 7-moxa-cone group were reduced as compared with those before treatment separately (all P<0.01), PPT values were increased as compared with those before treatment (all P<0.01). But the differences were not statistically significant in the sham-moxibustion group as compared with those before treatment (all P>0.05). After treatment and in 1-month follow-up, NPQ scores and MPQ scores in the 3-moxa-cone group, the 5-moxa-cone group and the 7-moxa-cone group were lower as compared with the sham-moxibustion group separately (all P<0.05), PPT values were higher as compared with the sham-moxibustion group (all P<0.05). After treatment, the NPQ score and MPQ score in the 7-moxa-cone group were lower than the 3-moxa-cone group and PPT values was higher than the 3-moxa-cone group (all P<0.05). After treatment and in 1-month follow-up, the therapeutic effects in the 3-moxa-cone group, the 5-moxa-cone group and the 7-moxa-cone group were better than the sham-moxibustion group separately (all P<0.05). But, the therapeutic effects were not significantly different in comparison among the moxibusiton groups (P>0.05). CONCLUSION: The direct moxibustion therapy with different small-moxa-cones effectively relieves chronic neck pain. There is a trend of improvement of the therapeutic effects with increase of the numbers of moxa cones.