- Volume 69, Issue 11, 2019

A polyphasic study was undertaken to establish the taxonomic provenance of a rapidly growing Mycolicibacterium strain, CECT 8783T, recovered from the plant Stellera chamaejasme L. in Yunnan Province, China. Phylogenetic analyses based upon 16S rRNA and whole-genome sequences showed that the strain formed a distinct branch within the evolutionary radiation of the genus Mycolicibacterium . The strain was most closely related to Mycolicibacterium moriokaense DSM 44221T with 98.4 % 16S rRNA gene sequence similarity, but was distinguished readily from this taxon by a combination of chemotaxonomic and phenotypic features and by low average nucleotide identity and digital DNA–DNA hybridization values of 79.5 and 21.1 %, respectively. Consequently, the strain is considered, to represent a novel species of Mycolicibacterium for which the name Mycolicibacterium stellerae sp. nov is proposed; the type strain is I10A-01893T (=CECT 8783T=KCTC 19843T=DSM 45590T).

Two novel, Gram-stain-positive, non-motile, facultatively anaerobic, rod-shaped bacteria (strains 2129T and 2119) were isolated from the faeces of Tibetan antelopes (Pantholops hodgsonii) on the Qinghai-Tibet Plateau, PR China. The 16S rRNA gene sequences of the strains showed highest similarity values to Actinomyces timonensis DSM 23838T (92.9 and 92.8 %, respectively), and phylogenetic analysis based on 16S rRNA gene and genomic sequences indicated that strains 2129T and 2119 represent a new lineage. Strains 2129T and 2119 could ferment d-adonitol and d-xylose, but were unable to utilize d-mannose and d-melibiose nor produce esterase (C4) and proline arylamidase. The G+C contents of the two strains were both 69.0 mol%. Their genomes exhibited less than 40.4 % relatedness in DNA–DNA hybridization tests (below 70 % as the recommended threshold for new species) with all available genomes of the genus Actinomyces in the NCBI database. The major fatty acids of the two strains were C18 : 1ω9c and C16 : 0, and the major polar lipids were diphosphatidylglycerol, glycolipid, phosphatidylinositol, phosphatidyl inositol mannoside and phosphoglycolipid. Based on the results of genotypic, phenotypic and biochemical analyses, it is proposed that the two unidentified bacteria be classified as representing a novel species, Actinomyces lilanjuaniae sp. nov. The type strain is 2129T (=CGMCC 4.7483T=DSM 106426T).

A novel actinobacterium, designated strain NEAU-THZ27T, was isolated from soil collected from the Cornel peak in Jiaozuo, Henan Province, PR China and characterized using a polyphasic approach. Morphological and chemotaxonomic characteristics of the strain coincided with those of members of the genus Kribbella . The results of 16S rRNA gene sequence analysis showed that strain NEAU-THZ27T belongs to the genus Kribbella and was most closely related to Kribbella podocarpi YPL1T (98.96 %), Kribbella karoonensis Q41T (98.89 %), Kribbella aluminosa HKI 0478T (98.86%) and Kribbella hippodromi S1.4T (98.85 %), similarities to other type strains of species of the genus Kribbella were found to be less than 98.7 %. Phylogenetic analyses using the 16S rRNA gene sequence and multilocus sequence analysis using the concatenated gene sequences of the gyrB, rpoB, recA, relA and atpD genes all showed that the strain formed a separate branch in the genus Kribbella . The cell wall contained ll-diaminopimelic acid as the major diamino acid and the whole-cell hydrolysates were ribose and glucose. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylinositol. The predominant menaquinone was MK-9(H4). Major fatty acids were iso-C16 : 0, iso-C14 : 0 and anteiso-C15 : 0, these chemotaxonomic data supported the affiliation of strain NEAU-THZ27T to the genus Kribbella . The DNA G+C content was 68.0 mol%. Furthermore, the strain could be clearly distinguished by concatenated gene genetic distances, the combination of DNA–DNA hybridization results and some phenotypic characteristics. Therefore, it is proposed that strain NEAU-THZ27T represents a novel species of the genus Kribbella , for which the name Kribbella jiaozuonensis sp. nov. is proposed. The type strain is NEAU-THZ27T (=CGMCC 4.7504T=DSM 105535T).

Arthrobacterenclensis was reported to cluster with species of the genus Pseudarthrobacter but the peptidoglycan containing lysine, alanine and glutamic acid and the presence of minor amounts of menaquinone MK-8(H4) were not in line with the description of the genus. Re-analysis of these traits revealed a peptidoglycan with l-Lys–l-Ser–l-Thr–l-Ala and no MK-8(H4), but major amounts of MK-9(H2) in the quinone system of A. enclensis DSM 25279T. These data demonstrate that A. enclensis shares the characteristics of the genus Pseudarthrobacter . Since the reported quinone systems of Pseudarthrobacterphenanthrenivorans [MK-8 and MK-9(H2)] and Pseudarthrobacterscleromae [MK-8(H2] were clearly different from those of other species of the genus, the quinone systems of the two species were re-analyzed. Since the polar lipid profile of P. phenanthrenivorans was reported to contain phosphatidylethanolamine, which is unusual for a member of the Micrococcaceae , and the polar lipid profile of P. scleromae was unknown, the polar lipids of these two species were also analysed. The quinone system of P. phenanthrenivorans DSM 18606T was composed of the major menaquinones MK-9(H2), MK-8(H2) and MK-10(H2) and that of P. scleromae DSM 17756T was composed of the major menaquinones MK-9(H2) and MK-8(H2) and MK-9. In the polar lipid profile of P. phenanthrenivorans DSM 18606T no phosphatidylethanolamine could be detected. Based on these results we here propose the reclassification of A. enclensis as Pseudarthrobacter enclensis comb. nov. and emend the descriptions of the genus Pseudarthrobacter and the two species P. phenanthrenivorans and P. scleromae .

Micrococcus aloeverae, Micrococcus endophyticus , Micrococcus luteus and Micrococcus yunnanensis are phenotypically and genotypically closely related, and together comprise the M. luteus group. In this study, the taxonomic relationships among Micrococcus aloeverae , M. luteus and M. yunnanensis were re-evaluated by using polyphasic approaches. The similarity values of the concatenated housekeeping gene (gyrB, recA and rpoB) sequences shared by the type strains of M. aloeverae , M. luteus and M. yunnanensis ranged from 98.3 to 99.4 %. The average nucleotide identity, average amino acid identity and digital DNA‒DNA hybridization values among these three taxa were greater (97.1‒98.1 %, 96.8‒98.1 % and 75.0‒83.5 %, respectively) than the thresholds for bacterial species delineation, indicating that they belong to the same species, whereas those for M. endophyticus were clearly lower than the thresholds. In addition, phenotypic and chemotaxonomic characterization results also support the synonymy of these three taxa. Therefore, we propose that M. aloeverae and M. yunnanensis should be reclassified as later heterotypic synonyms of M. luteus .

A Gram-stain-positive, rod-shaped and motile bacterium with lateral flagellum, designated T3246-1T, was isolated from an ice core, which was drilled from Hariqin Glacier on the Tibetan Plateau, PR China. It grew optimally at 20 °C, pH 7–8 and in the presence of 3 % (w/v) NaCl. The major fatty acid of strain T3246-1T was anteiso-C15 : 0. Major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol. MK-8 was the dominant isoprenoid quinone. The whole-cell sugars were rhamnose, xylose and mannose. The major cell-wall peptidoglycan was lysine. The genomic DNA G+C content of the strain was 71.4 mol%. Results of phylogenetic analysis based on 16S rRNA gene sequences showed that strain T3246-1T formed a lineage within the genus Haloactinobacterium and was closely related to Haloactinobacterium album YIM 93306T with 95.99 % similarity. The average nucleotide identity value between strain T3246-1T and Haloactinobacterium album YIM 93306T was 76.65 %. Based on phenotypic and chemotaxonomic characteristics, strain T3246-1T was considered to represent a novel species of the genus Haloactinobacterium , for which the name Haloactinobacterium glacieicola sp. nov. is proposed. The type strain is T3246-1T (=CGMCC 1.13535T=JCM 32923T).

A novel actinomycete, designated strain GLM-1T, was isolated from arbuscular mycorrhizal fungal spores from Funneliformis mosseae RYA08, collected from Aquilaria crassna Pierre ex Lec. rhizosphere soil in Klaeng, Rayong Province, Thailand. Morphological characteristics of this strain included long chains of rod-like cells and squarish elements. The cell-wall composition of this novel isolate contained meso-diaminopimelic acid. The whole-cell diagnostic sugars were arabinose and galactose. The predominant menaquinone was MK-9(H4). The major fatty acids were iso-C16 : 0 and iso-C15 : 0. Only phosphatidylethanolamine was detected as a polar lipid. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GLM-1T was closely related to   Amycolatopsis rhabdoformis   SB026T (99.11 %) with a low DNA–DNA hybridization value of 22.6–34.7 %. Genome sequencing revealed a genome size of 10 Mbp. There were obvious distinctions in the average nucleotide identity values between stain GLM-1T and its closely related strains at around 86–93 % (ANIb) and 89–94 % (ANIm). The digital DNA–DNA hybridization values between strain GLM-1T and type strains of phylogenetically related species were 34–55 %. The G+C content of the genomic DNA was 71.8 mol%. Based on these data, strain GLM-1T is considered to represent a novel species of the genus   Amycolatopsis  , for which the name   Amycolatopsis eburnea sp. nov. is proposed. The type strain is GLM-1T (=TBRC 9315T=NBRC 113658T)

Two Gram-stain-positive, irregular rod-shaped (0.2–0.5×1.3–2.5 um) strains, HY056T and HY057, were isolated from the faeces of Equus kiang (the largest of the wild asses) collected at different regions from the Qinghai-Tibetan Plateau of PR China. Phylogenetic analyses indicated that strains HY056T and HY057 belong to the genus Nocardioides by sharing a similarity ranging from 96.3 to 97.0 % in the 16S rRNA gene sequence and forming a distinct cluster with Nocardioides daphniae JCM 16608T, Nocardioides houyundeii 78T, Nocardioides solisilvae Ka25T and Nocardioides gilvus XZ17T. The digital DNA–DNA hybridization value of strain HY056T was 96.9 % with strain HY057, but less than 30.0 % with the above four closest relatives. MK-8(H4) was the predominant (91.6 %) respiratory quinone. The cell wall contained ll-2,6-diaminopimelic acid as the diamino acid of the peptidoglycan. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phospholipid. The predominant fatty acids (>10.0 %) were C18 : 1ω9c and iso-C16:0. The DNA G+C contents of strains HY056T and HY057 were 68.9 and 69.1 mol%, respectively. β-Glucosidase expression was positive, and acid was produced from d-fructose. Strain HY056T (=CGMCC 4.7563T=JCM 33399T) is assigned as the type strain of a novel species within the genus Nocardioides , for which the name Nocardioides yefusunii sp. nov is proposed.

An extremely halophilic archaeon, strain F13-25T, was isolated from a marine saltern located in Isla Cristina, Huelva, on the south-west coast of Spain. The novel strain had pink-pigmented, non-motile, coccoid cells. Optimal growth was achieved at 25 % (w/v) NaCl, pH 7.5 and 37 °C. Strain F13-25T possessed two heterogeneous 16S rRNA genes (rrnA and rrnB) most closely related to Halorientalis persicus D108T (97.6–99.2 % sequence similarity) and Halorientalis regularis TNN28T (95.9–98.8 %). On the basis of the results of rpoB′ gene sequence analysis, strain F13-25T was also closely related to Halorientalis persicus IBRC-M 10043T (89.9 %) and Halorientalis regularis TNN28T (92.3 %). Relatedness values, computed using the Genome-to-Genome Distance Calculator, between strain F13-25T and Halorientalis persicus IBRC-M 10043T and Halorientalis regularis IBRC-M 10760T were 34.6 and 36.2 %, respectively. Average nucleotide identity values based on orthoANI, ANIb and ANIm of strain F13-25T and Halorientalis persicus IBRC-M 10043T and Halorientalisregularis IBRC-M 10760T were 88.0 and 88.8, 87.1 and 87.6 %, and 89.2 and 89.6 %, respectively. All values were far below the threshold accepted for prokaryotic species delineation. The major polar lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and one glycolipid chromatographically identical to sulfated diglycosyl diether. The DNA G+C content was 65.7 mol% (genome). The results of phylogenetic, phenotypic and chemotaxonomic analyses indicated that strain F13-25T represents a novel species of the genus Halorientalis , for which the name Halorientalis pallida sp. nov., with type strain F13-25T (=CECT 9384T=IBRC-M 11176T), is proposed.

In a survey of endophytic bacteria in Miscanthus sinensis, a strain of Gram-negative, non-endospore-forming, rod-shaped, aerobic bacterium was isolated and designated as RS10T. Phylogenetic analysis based on its 16S rRNA gene sequence showed that the strain RS10T was affiliated with the genus Pedobacter and exhibited the highest sequence similarities to Pedobacter kyungheensis KACC 16221T (97.78 %), Pedobacter roseus KCTC 22187T (97.75 %), Pedobacter humicola KACC 18452T (97.29 %) and Pedobacter soli KACC 14939T (97.23 %). The novel strain contained iso-C15 : 0, iso-C17 : 0-3OH and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as major fatty acids. The only isoprenoid quinone detected in strain RS10T was MK-7. Strain RS10T contains phosphatidylethanolamine and one kind of aminophospholipid as its major polar lipids. The DNA G+C content for RS10T was 39.8 mol%. Based on the results of phenotypic and genomic characterizations, we concluded that strains RS10T represents a novel species of Pedobacter , for which the name Pedobacter miscanthi sp. nov. is proposed. The type strain is RS10T (=KCTC 62786T=GDMCC 1.1415T).

A bacterial strain M05W1-28T was isolated from a well that collected water for irrigation from a deep aquifer at a depth of 400 m. Cells were observed to be rod-shaped, non-motile, aerobic, stained Gram-negative. Optimal growth was obtained at pH 7.0 (range: 6.0–9.0), 28 °C (range: 15–37 °C) and 0 % NaCl (range: 0–1.5 %, w/v) in modified tryptic soy broth (mTSB) without added NaCl and R2A. The cells were found to be positive for catalase and oxidase activities. The major fatty acids (>10 %) were identified as summed feature 3 (C16 : 1  ω7c / C16 : 1  ω6c) and iso-C15 : 0. The major polar lipids were phosphatidylethanolamine, glycolipid, phosphoglycolipids, phospholipids, and unidentified lipids. The major respiratory quinone was menaquinone-7 (MK-7). The genomic G+C content of strain M05W1-28T was 40.7 %. Based on similarities of 16S rRNA gene sequences, strain M05W1-28T was affiliated with the genus Sphingobacterium , exhibiting the highest sequence similarities with S. multivorum LMG 8342T (97.5 %), S. ginsenosidimutans THG07T (97.1 %) and less than 97.0 % to other members of the genus. The average nucleotide identity (ANI) and digital DNA–DNA hybridisation values (dDDH) between M05W1-28T and S. multivorum LMG 8342T were 78.1 and 22.5 %, respectively. Phenotypic characteristics including enzyme activities and carbon source utilisation differentiated the strain from other Sphingobacterium species. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain M05W1-28T represented a novel species within the genus Sphingobacterium , for which the name Sphingobacterium puteale sp. nov. is proposed. The type strain is M05W1-28T (=CGMCC 1.13711T=KCTC 72027T).

A Gram-stain-negative bacterial strain, designated CA10T, was isolated from bovine raw milk sampled in Anseong, Republic of Korea. Cells were yellow-pigmented, aerobic, non-motile bacilli and grew optimally at 30 °C and pH 7.0 on tryptic soy agar without supplementation of NaCl. Phylogenetic analysis based on the 16S rRNA gene sequences revealed that strain CA10T belonged to the genus Chryseobacterium , family Flavobacteriaceae , and was most closely related to Chryseobacterium indoltheticum ATCC 27950T (98.75 % similarity). The average nucleotide identity and digital DNA–DNA hybridization values of strain CA10T were 94.4 and 56.9 %, respectively, relative to Chryseobacterium scophthalmum DSM 16779T, being lower than the cut-off values of 95–96 and 70 %, respectively. The predominant respiratory quinone was menaquinone-6; major polar lipid, phosphatidylethanolamine; major fatty acids, iso-C15 : 0, summed feature 9 (iso-C17 : 1ω9c and/or C16 : 0 10-methyl), summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1ω7c) and iso-C17 : 0 3-OH. The results of physiological, chemotaxonomic and biochemical analyses suggested that strain CA10T is a novel species of genus Chryseobacterium , for which the name Chryseobacterium mulctrae sp. nov. is proposed. The type strain is CA10T (=KACC 21234T=JCM 33443T).

A bacterial strain, 17J42-9T, was isolated from a soil sample collected on Jeju Island, Republic of Korea. Cells were observed to be Gram-stain negative and rod-shaped. Colonies were observed to be orange in colour on R2A agar. Analysis of 16S rRNA gene sequences showed that high levels of 16S rRNA sequence similarity were shared between 17J42-9T and Emticicia fontis IMCC1731T (98.2 %), Emticicia ginsengisoli Gsoil 085T (98.2 %) and Emticicia soli ZZ-4T (97.8 %). Growth of strain 17J42-9T was observed at 10–37 °C, pH 6.0–8.5 and in the presence of 0–0.5 % NaCl. The genomic G+C content was calculated to be 38.6 mol%. The predominant respiratory quinone of the isolate was found to be MK-7; the major fatty acids were identified as summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) (34.1 %), C15 : 0iso (23.4 %) and C17 : 0iso 3-OH (10.8 %). The major polar lipids were found to be phosphatidylethanolamine, two unidentified aminolipids and an unidentified lipid. The phenotypic and chemotaxonomic data support the affiliation of strain 17J42-9T with the genus Emticicia . However, the DNA–DNA relatedness between the isolate and its closest phylogenetic neighbours was lower than 46 %. The results of 16S rRNA gene sequence similarity analysis, DNA–DNA hybridization analysis and the observed differentiating phenotypic properties from other closely related taxa clearly indicate that strain 17J42-9T represents a novel species in the genus Emticicia , for which the name Emticicia agri sp. nov. is proposed. The type strain is 17J42-9T (=KCTC 62270T=JCM 33056T).

A Gram-stain-negative, rod-shaped and red-pigmented strain, HME7025T, was isolated from freshwater sampled in the Republic of Korea. Phylogenetic analysis based on its 16S rRNA gene sequence revealed that strain HME7025T formed a lineage within the family Cytophagaceae of the phylum Bacteroidetes . Strain HME7025T was closely related to the genera Pseudarcicella , Arcicella and Flectobacillus . The 16S rRNA gene sequence similarity values of strain HME7025T were under 94.5 % to its closest phylogenetic neighbours. The major fatty acids of strain HME7025T were iso-C15 : 0 (41.9 %), summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c; 12.2 %) and anteiso-C15 : 0 (10.8 %). The major respiratory quinone was menaquinone-7. The major polar lipids were phosphatidylethanolamine, two unidentified aminophospholipids and one unidentified polar lipid. The DNA G+C content of strain HME7025T was 37.9 mol%. On the basis of the evidence presented in this study, strain HME7025T represents a novel species of a novel genus within the family Cytophagaceae , for which the name Allopseudarcicella aquatilis gen. nov., sp. nov. is proposed. The type strain is HME7025T (=KCTC 23617T=CECT 7957T).

A novel marine Gram-stain-negative, non-motile, aerobic and rod-shaped bacterium, designated strain DSL-48T, was isolated from tidal flat sediment sampled from the East China Sea and characterized phylogenetically and phenotypically. Bacterial optimal growth occurred at 35 °C (range, 4–37 °C), pH 6 (pH 5–10) and with 4 % (w/v) NaCl (0–7 %). The nearest phylogenetic neighbour was Confluentibacter citreus KCTC 52638T with 16S rRNA gene similarity of 97.1 %. The predominant respiratory quinone was menaquinone-6 (MK-6). The major polar lipids were phosphatidylethanolamine, three unidentified aminolipids and four unidentified lipids. The major fatty acids of strain DSL-48T were iso-C15 : 0, iso-C17 : 0 3-OH, anteiso-C15 : 0, iso-C15 : 0 3-OH and iso-C16 : 0 3-OH. The G+C content of the genomic DNA was 33.3 mol%. The combined genotypic and phenotypic data indicated that strain DSL-48T represents a novel species of the genus Confluentibacter , for which the name Confluentibacter sediminis sp. nov. is proposed, with the type strain DSL-48T (=KCTC 62648T=MCCC 1K03537T).

We describe the isolation and characterization of three bacterial isolates from the common house fly, Musca domestica, caught in Londerzeel, Belgium and Huye District, Rwanda. Although isolated from distinct geographical locations, the strains show >99 % identical 16S rRNA gene sequences and are <95 % identical to type strains of Apibacter species. Whole-genome sequences were obtained for all three strains. The genomes are 2.4–2.5 Mb with a G+C content of ~30.3 mol%. Bacteriological and biochemical analysis of the strains demonstrate distinctly different characteristics compared to known Apibacter species. Particularly, the three strains investigated in this study can be distinguished from the known Apibacter species ( Apibacter mensalis and Apibacter adventoris ) through urease and β-glucosidase activities. Whole-cell fatty acid methyl ester analysis shows that the fatty acid composition of the novel strains is also unique. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify these isolates as representatives of a novel species of the genus Apibacter , Apibacter muscae sp. nov., in reference to its prevalence in house flies, with strain G8T (=LMG 30898T=DSM 107922T) as the type strain.

A Gram-stain-negative, rod-shaped, red-coloured and aerobic bacterium, designated 2b14T, was isolated from rhizosphere soil of Saxifraga oppositifolia sampled at the Chinese Arctic Yellow River Station in Norway. Optimal growth occurred at 28 °C (range, 4–37 °C) and pH 7.0–7.5 (pH 6.5–8.5). The strain could tolerate up to 2.5 % (w/v) NaCl concentration. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain 2b14Thad the highest similarity value of 96.0 % to Pontibacter diazotrophicus CCTCC AB 2013049T. The major fatty acids were summed feature 4 (anteiso-C17 : 1 B and/or iso-C17 : 1 I), iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipid was found to be phosphatidylethanolamine. The genomic DNA G+C content of strain 2b14T was 45.5 mol%. The sole respiratory quinone was MK-7. The combined results of physiological, genotypic, phylogenetic and chemotaxonomic analyses showed that strain 2b14T represents a novel species within the genus Pontibacter, for which the name Pontibacter arcticus sp. nov. is proposed. The type strain is 2b14T (=KCTC 62596T=MCCC 1H00304T).