- Volume 69, Issue 9, 2019

An actinomycete strain, TKZ-21T, was isolated from a freshwater alga (Chetophoraceae) collected from the Takizawa River, Yamanashi, Japan, and examined using a polyphasic taxonomic approach. Cells were Gram-stain positive, aerobic, non-sporulating, motile, and coccoid or short rod-shaped. The strain grew in the presence of 0–4 % (w/v) NaCl, between pH 6–9.4, and over a temperature range of 15–40 °C, with optimum growth at 30 °C. The peptidoglycan type of strain TKZ-21T was A4β, containing l-ornithine as diagnostic diamino acid and d-glutamic acid as the interpeptide bridge. The predominant menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, ninhydrin-positive glycolipid, and unidentified phospholipids. The major cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0, and the DNA G+C content was 75.6 mol%. On the basis of 16S rRNA gene sequence analysis, strain TKZ-21T was closely related to Cellulomonas fimi (98.5 % sequence similarity) and Cellulomonas biazotea (98.3 %). The genome orthoANI value between strain TKZ-21T and C. biazotea and C. fimi were 84.7 and 84.2 %, respectively. On the basis of fatty acid and MALDI-TOF MS profile analysis, phylogenetic analyses, genomic analysis, and phenotypic data, it is proposed that the isolate be classified as a representative of a novel species of the genus Cellulomonas , with the name Cellulomonas algicola sp. nov. The type strain is TKZ-21T (=NBRC 112905T=TBRC 8129T).

A novel actinobacterial strain, designated Z022T, isolated from a soil sample collected from Dangxiong in Tibet Autonomous Region (PR China), was determined by polyphasic taxonomic approach. The organism had chemotaxonomic and morphological properties consistent with its classification in the genus Streptomyces . Strain Z022T showed high similarity value to Streptomyces lucensis NBRC 13056T (98.87 %) and S. achromogenes subsp . achromogenes NBRC 12735T (98.68 %) based on the 16S rRNA gene phylogenetic tree. The genomic DNA G+C content of strain Z022T based on the genome sequence was 72.16 mol%. DNA–DNA relatedness values between strain Z022T and strain Streptomyces lucensis NBRC 13056T was 23.7±1.3 % and significantly lower than 70 %. Chemotaxonomic data revealed that strain Z022T possessed MK-9(H8) and MK-9(H6) as the predominant menaquinone, ll-diaminopimelic acid as the diagnostic diamino acid, and galactose, glucose, xylose and ribose as whole cell sugars. Diphosphatidylglycerol (DPG) and phosphatidylethanolamine (PE) were the predominant polar lipids; anteiso-C15 : 0, iso-C16 : 0, and anteiso-C17 : 0 were the major fatty acids. On the basis of these genotypic and phenotypic data, it is proposed that isolate Z022T (=JCM 31053T=CGMCC 4.7273T) should be classified in the genus Streptomyces as Streptomyces dangxiongensis sp. nov.

Four Gram-stain positive, rod-shaped bacterial isolates, strains JZ R-183T, JZ RK-117, DI-46 and JZ R-35T, were recovered from bulk tank raw cow’s milk from three different dairy farms in Germany. Analysis of their 16S rRNA gene sequences indicated that these isolates belonged to the family Micrococcaceae , closely related to the genera Arthrobacter , Neomicrococcus, Glutamicibacter and Citricoccus . The 16S rRNA gene sequence similarity between the isolates and the next related type strains was below 97.3 %. Phylogenetic analysis of 16S rRNA, recA and gyrB genes revealed that these isolates formed two different groups in an independent cluster within the family Micrococcaceae . Chemotaxonomic analyses determined anteiso-C15 : 0 as predominant fatty acid, but also large amounts of iso-C15 : 0, iso-C16 : 0 and iso-C17 : 0 were detected. The menaquinones MK-9(H2) and MK-7(H2) were present in all of the isolates and the polar lipid pattern contained the phospholipids diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol and a glycolipid. The peptidoglycan type of the isolates was A4α, with alanine, lysine and glutamate as dominating cell wall amino acids. The fatty acid and menaquinone profile differentiated the strains from the genera Arthrobacter , Neomicrococcus, Citricoccus and Glutamicibacter. The results of phylogenetic, phenotypic and chemotaxonomic analyses indicated that the isolates belonged to two novel species of a novel genus, for which the names Galactobacter caseinivorans gen. nov., sp. nov. and Galactobacter valiniphilus sp. nov. are proposed. The type strains are JZ R-183T (=DSM 107700T=LMG 30902T) and JZ R-35T (=DSM 107699T=LMG 30901T).

A novel endophytic actinomycete strain AZ1-13T was isolated from roots of Azadirachta indica, and its taxonomic position was investigated using a polyphasic approach. Pairwise 16S rRNA gene sequence similarities of strain AZ1-13T and its closest species, Jishegella zingiberis PLAI1-1T and Micromonospora endophytica 202201T, were 99.7 and 99.2 %, respectively. Phylogenetic analyses of the family Micromonosporaceae based on 16S rRNA gene sequences indicated strains AZ1-13T and J. zingiberis PLAI1-1Tare located within the genus Micromonospora . The approximate genome size of the strain was 5.96 Mb with 71.9 mol% of G+C content. The strain AZ1-13T exhibited ANIb values of 87.4 % with J. zingiberis PLAI1-1T and 85.1 % with M. endophytica 202201T. Chemotaxonomic characteristics of strain AZ1-13T were consistent within the genus Micromonospora : cell-wall peptidoglycan of the strain contained meso-diaminopimelic acid; glucose, mannose, ribose and xylose are presented as the whole-cell sugars; the predominant menaquinones were MK-9(H4) and MK-9(H6); major cellular fatty acids were iso-C15 : 0, 10-methyl C17 : 0, C17 : 0, anteiso-C17 : 0 and iso-C17 : 1ω8c; diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol were detected as distinguished phospholipids. Based on phenotypic properties, phylogeny and genomic data, the strain AZ1-13T could be distinguished from its closest neighbours, representing a novel species of the genus Micromonospora , for which the name Micromonospora radicis sp. nov. is proposed. The type strain is AZ1-13T (=KCTC 39786T=NBRC 112324T=JCM 32147T = TISTR 2404T). This study also proposed that J. zingiberis is transferred to the genus Micromonospora as Micromonospora zingiberis comb. nov. (type strain PLAI1-1T=TBRC 7644T=NBRC 113144T=JCM 32592T).

One novel actinobacterial isolate, designated strain NEAU-G17T, was isolated from muddy soil collected from a riverbank in Chad and characterised using polyphasic approach. Morphological and chemotaxonomic characteristics of this strain coincided with those of the genus Actinomadura . The 16S rRNA gene sequence analysis showed that strain NEAU-G17T was most closely related to Actinomadura oligospora JCM 10648T (98.7 %) and Actinomadura gamaensis NEAU-Gz5T (98.6 %). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NEAU-G17T formed a cluster with Actinomadura oligospora JCM 10648T, Actinomadura gamaensis NEAU-Gz5T and Actinomadura rupiterrae CS5-AC15T (96.4 %). However, the combination of DNA–DNA hybridisation values and some phenotypic characteristics allowed the isolate to be differentiated from their most closely related species. The major menaquinones were identified as MK-9(H4), MK-9(H6) and MK-9(H8). Their polar lipid profile was found to contain diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, an unidentified phospholipid and an unidentified lipid. The major fatty acids were identified as C16 : 0, iso-C16 : 0 and C18 : 1 ω9c. Therefore, it is proposed that strain NEAU-G17T should be classified as representative of a novel species of the genus Actinomadura , for which the name Actinomadura logoneensis sp. nov. is proposed. The type strains is NEAU-G17T (=CGMCC 4.7411T=DSM 105122T).

Three Gram-stain-positive, rod-to-coccoid-shaped, catalase-positive and non-motile bacterial strains isolated from the choanae of a Northern bald ibis, designated strains 200CHT, W8T and 812CHT, respectively, were subjected to comprehensive taxonomic characterization. The three strains were oxidase-negative. The 16S rRNA gene sequence of 200CHT showed highest similarities to Corynebacterium epidermidicanis 410T (96.7 %) followed by Corynebacterium argentoratense DSM 44202T, Corynebacterium ulcerans NCTC 7910T and Corynebacterium pseudotuberculosis CIP 102968T (each 96.3 %). Strains W8T and 812CHT both showed highest 16S rRNA gene sequence similarities to Corynebacterium pelargi 136/3T (98.0 and 99.9 %, respectively). Comparison of the partial housekeeping gene sequence of fusA showed higher sequence similarities of 812CHT to C. pelargi (95.8 %) than W8T (90.9 %) which was also confirmed by corresponding amino acid sequences. In both, fusA gene and corresponding protein sequence strain 200CHT showed low sequence similarities to C. epidermidicanis 410T(81.6 and 87.4 %, respectively). Strains 812CHT and W8T had 76.7 % ANI similarity to each other and 88.2 and 76.4 % to C. pelargi 136/3T, respectively. In silico DNA–DNA hybridization values for 812CHT and W8T were 22.1 % among the two strains and 35.3 and 21.7 % to C. pelargi 136/3T, respectively. These data not only demonstrate that strain W8T is a representative of a novel species, but despite the high 16S rRNA gene sequence similarity to C. pelargi , strain 812CHT is also a representative of another novel species. All three strains possessed corynemycolic acids and contained meso-diaminopimelic acid as the diagnostic diamino acid of the peptidoglycan. The two strains, 200CHT and W8T, are distinguished from each other and established Corynebacterium species phylogenetically and phenotypically. In conclusion, three novel species of the genus Corynebacterium are proposed, namely Corynebacterium pseudopelargi 812CHT (=LMG 30627T=CCM 8832T), Corynebacterium choanae 200CHT (=LMG 30628T=CCM 8831T) and Corynebacterium gerontici W8T (=LMG 30629T=CCM 8833T), respectively.

Two hitherto unknown bacteria (strains 313T and 352) were recovered from the faeces of Tibetan antelopes on the Tibet–Qinghai Plateau, PR China. Cells were rod-shaped and Gram-stain-positive. The optimal growth conditions were at 37 °C and pH 7. The isolates were closely related to Actinotignum sanguinis (92.6 % 16S rRNA gene sequence similarity), Arcanobacterium haemolyticum (92.5 %), Actinotignum schaalii (92.4 %), Actinobaculum massiliense (92.2 %) and Flaviflexus huanghaiensis (91.6 %). Phylogenetic analyses showed that strains 313T and 352 clustered independently in the vicinity of the genera Actinotignum , Actinobaculum and Flaviflexus , but could not be classified clearly as a member of any of these genera. Phylogenomic analysis also indicated that strains 313T and 352 formed an independent branch in the family Actinomycetaceae . The major cellular fatty acids of the strains were C16 : 0 and C18 : 1ω9c. The polar lipids comprised diphosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylglycerol, phosphatidylinositol and five unidentified components. The peptidoglycan contained lysine, alanine and glutamic acid. The respiratory quinone was absent. The whole-cell sugars included glucose and rhamnose. The DNA G+C content of strain 313T was 60.6 mol%. Based on the low 16S rRNA gene sequence similarities, its taxonomic position in the phylogenetic and phylogenomic trees and its unique lipid pattern, we propose that strains 313T and 352 represent members of a novel species in a new genus, for which the name Fudania jinshanensis gen. nov., sp. nov. is proposed. The type strain is 313T (=CGMCC 4.7453T=DSM 106216T).

Eight pure cultures of alkaliphilic haloaloarchaea capable of growth by dissimilatory sulfur reduction (previously only shown for neutrophilic haloarchaea) were isolated from hypersaline alkaline lakes in different geographic locations. These anaerobic enrichments, inoculated with sediments and brines, used formate, butyrate and peptone as electron donors and elemental sulfur as an electron acceptor 4 M total Na+ and at pH 9–10. According to 16S rRNA gene sequencing, the isolates fell into two distinct groups. A major group, comprising seven obligate alkaliphilic isolates from highly alkaline soda lakes, represents a new species-level branch within the genus Natronolimnobius (order Natrialbales ), while a single moderately alkaliphilic isolate from the less alkaline Searles Lake forms a novel genus-level lineage within the order Haloferacales . The cells of the isolates are either flat rods or coccoid. They are facultative anaerobes using formate or H2 (in the presence of acetate or yeast extract as carbon source), C4–C9 fatty acids or peptone (the major group) as electron donors and either sulfur or DMSO (the major group) as electron acceptors. Aerobic growth is only possible with organic acids and peptone–yeast extract. All isolates are extreme halophiles, growing optimally at 4 M total Na+. On the basis of their unique physiological properties and distinct phylogeny, we propose that the seven isolates from the soda lakes are placed into a novel species, Natronolimnobius sulfurireducens sp. nov. (type strain AArc1T=JCM 30663T=UNIQEM U932T), and the Searles Lake isolate, AArc-SlT, into a new genus and species Halalkaliarchaeum desulfuricum (=JCM 30664T=UNIQEM U999T).

Three bacterial strains, 30S-ANTBAC, 103A-SOEBACH and 59G- WUEMPEL, were isolated from two small freshwater creeks and an intermittent pond near Salzburg, Austria. Phylogenetic reconstructions with 16S rRNA gene sequences and, genome based, with amino acid sequences obtained from 119 single copy genes showed that the three strains represent a new genus of the family Cytophagaceae within a clade formed by the genera Pseudarcicella , Arcicella and Flectobacillus . blast searches suggested that the new genus comprises widespread freshwater bacteria. Phenotypic, chemotaxonomic and genomic traits were investigated. Cells were rod shaped and were able to glide on soft agar. All strains grew chemoorganotrophically and aerobically, were able to assimilate pectin and showed an intense red pigmentation putatively due to various carotenoids. Two strains possessed genes putatively encoding proteorhodopsin and retinal biosynthesis. Genome sequencing revealed genome sizes between 2.5 and 3.1 Mbp and G+C contents between 38.0 and 42.7 mol%. For the new genus we propose the name Aquirufa gen. nov. Pairwise-determined whole-genome average nucleotide identity values suggested that the three strains represent two new species within the new genus for which we propose the names Aquirufa antheringensis sp. nov. for strain 30S-ANTBACT (=JCM 32977T =LMG 31079T=DSM 108553T) as type species of the genus, to which also belongs strain 103A-SOEBACH (=DSM 108555=LMG 31082) and Aquirufa nivalisilvae sp. nov. for strain 59G-WUEMPELT (=LMG 31081T =DSM 108554T).

A Gram-stain-negative, aerobic, non-motile and rod-shaped or filamentous bacterial strain, designated SHSM-M15T, was isolated from a salt marsh at Siheung in Republic of Korea and identified by polyphasic taxonomic study. Strain SHSM-M15T grew optimally at 25 °C, at pH 7.0–8.0 and in the presence of 1.0–2.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences showed that strain SHSM-M15T clusters with the type strain of Ancylomarina subtilis , showing 16S rRNA gene sequence similarity of 97.8 %. Strain SHSM-M15T had 16S rRNA gene sequence similarities of less than 93.7 % with the type strains of other recognised species. Strain SHSM-M15T contained MK-7 as the predominant menaquinone and iso-C15 : 0, iso-C15 : 0 3-OH and anteiso-C15 : 0 as the major fatty acids. The major polar lipid detected in strain SHSM-M15T was phosphatidylethanolamine. The DNA G+C content of strain SHSM-M15T from genomic sequence was 36.6 %. Mean DNA–DNA relatedness value between strain SHSM-M15T and the type strain of A. subtilis was 18 % and the average nucleotide identity value between strain SHSM-M15T and the type strain of A. subtilis was 87.98 %. The phylogenetic and genetic data and differential phenotypic properties indicated that strain SHSM-M15T is separated from A. subtilis . On the basis of the polyphasic data presented, strain SHSM-M15T is considered to represent a novel species of the genus Ancylomarina , for which the name Ancylomarina salipaludis sp. nov. is proposed. The type strain is SHSM-M15T (=KACC 19862T=NBRC 113749T).

A set of three psychrotrophic bacterial strains was isolated from different soil samples collected at the deglaciated northern part of James Ross Island (Antarctica) in 2014. All isolates were rod-shaped, Gram-stain-negative, non-motile, catalase-positive and oxidase-negative, and produced moderately slimy red-pink pigmented colonies on Reasoner's 2A (R2A) agar. A polyphasic taxonomic approach based on 16S rRNA gene sequencing, whole-genome sequencing, automated ribotyping, MALDI-TOF MS, chemotaxonomy methods and extensive biotyping using conventional tests and commercial identification kits was applied to the isolates in order to clarify their taxonomic position. Phylogenetic analysis based on the 16S rRNA gene showed that all isolates belonged to the genus Hymenobacter with the closest relative being Hymenobacter aerophilus DSM 13606T, exhibiting 98.5 % 16S rRNA gene pairwise similarity to the reference isolate P6312T. Average nucleotide identity values calculated from the whole-genome sequencing data proved that P6312T represents a distinct Hymenobacter species. The major components of the cellular fatty acid composition were summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c), C16 : 1 ω5c, summed feature 4 (C17 : 1 anteiso B/iso I), C15 : 0 anteiso and C15 : 0 iso. The menaquinone system of strain P6312T contained MK-7 as the major respiratory quinone. The predominant polar lipids were phosphatidylethanolamine and an unidentified phospholipid. Moderate to minor amounts of three unidentified polar lipids, four unidentified aminophospholipids, one unidentified glycolipid and one unidentified phospholipid were also present. Based on the obtained results, we propose a novel species for which the name Hymenobacter humicola sp. nov. is suggested, with the type strain P6312T (=CCM 8763T=LMG 30612T).

During a study of the Kyonggi University soil bacterial diversity, an aerobic, non-motile, Gram-stain-negative, non-spore-forming, rod-shaped, yellow pigmented bacterium, designated strain RD-2-33T was isolated. Strain RD-2-33T grew optimally at 28–35 °C and pH 7.0–7.5; hydrolysed gelatin and DNA; and tolerated 1 % of NaCl. A phylogenetic analysis based on its 16S rRNA gene sequence revealed that strain RD-2-33T clustered with the genus Flavobacterium . The closest member was Flavobacterium dankookense ARSA-19T (97.1 % sequence similarity) and Flavobacterium cheonhonense ARSA-15T (96.7 %). Sole respiratory quinone was MK-6. The major polar lipids were phosphatidylethanolamine and an unidentified polar lipid. The major cellular fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, and iso-C15 : 1 G. The DNA G+C content was 38.6 mol%. The average nucleotide identity (ANI) and in silico DNA–DNA hybridisation relatedness between strain RD-2-33T and Flavobacterium dankookense DSM 25687T were 75.2 and 19.3 %, respectively. Based on the polyphasic and phylogenetic data, strain RD-2-33T represents a novel species of the genus Flavobacterium , for which the name Flavobacterium silvisoli sp. nov. is proposed. The type strain is RD-2-33T (=KEMB 9005–742T=KACC 21178T=NBRC 113789T).

A Gram-stain-negative, aerobic, non-motile and ovoid or rod-shaped bacterial strain, MRS2T, was isolated from an intestine of Nile tilapia (Oreochromisniloticus) collected from the Republic of Korea. Strain MRS2T grew optimally at 30 °C and in the presence of 0–2.0 % (w/v) NaCl. A neighbour-joining phylogenetic tree of 16S rRNA gene sequences showed that strain MRS2T clustered with the type strains of Empedobacter species. It exhibited the highest 16S rRNA gene sequence similarity (98.5 %) to the type strain of Empedobacter falsenii and sequence similarities of 97.4–97.6 % to the type strains of two other Empedobacter species. Strain MRS2T contained MK-6 as the predominant ubiquinone and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C17 : 0 3-OH and iso-C15 : 0 as the major fatty acids. The major polar lipids of strain MRS2T were phosphatidylethanolamine, one unidentified lipid and one unidentified aminolipid. The DNA G+C contents of strain MRS2T were 32.2 mol% or 30.65 mol%. Strain MRS2T exhibited DNA–DNA relatedness values of 12–20 % to the type strains of Empedobacter falsenii , Empedobacter brevis and Empedobacter stercoris . The average nucleotide identity values between strain MRS2T and five strains of E. falsenii and E. brevis were 84.8–91.0 %. The phylogenetic, genetic and differential phenotypic properties indicated that strain MRS2T is separated from Empedobacter species. On the basis of the data presented here, strain MRS2T is considered to represent a novel species of the genus Empedobacter , for which the name Empedobactertilapiae sp. nov. is proposed. The type strain is MRS2T (=KCTC 62904T=NBRC 113550T).

A Gram-stain-negative, aerobic, rod-shaped, non-motile, pink-pigmented bacterium, designated NLT, was isolated from arsenic-contaminated farmland soil. Strain NLT showed the highest 16S rRNA gene sequence similarities with those of Hymenobacter jeollabukensis 1-3-3-8T (98.9 %), Hymenobacter gummosus ANT-18T (97.5 %), Hymenobacter paludis KBP-30T (97.4 %), Hymenobacter ocellatus Myx2105T (97.1 %) and Hymenobacter coalescens WW84T (96.4 %). The values of genomic orthoANI and dDDH between strain NLT and Hymenobacter jeollabukensis KCTC 52741T was 90.5 and 41.2 %, respectively, and those between strain NLT and Hymenobacter gummosus KCTC 52166T was 84.4 and 28.4 %, respectively. Strain NLT exhibited DNA–DNA hybridisation values of 41.3 and 44.1 % with Hymenobacter paludis KCTC 32237T and Hymenobacter ocellatus DSM 11117T, respectively. Strain NLT had major fatty acids (>10 %) of summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B), iso-C15 : 0 and anteiso-C15 : 0 and the predominant polyamine of homospermidine. The only respiratory quinone was menaquinone-7. The polar lipids were phosphatidylethanolamine, phospholipid, three unidentified lipids and two amino lipids. Strain NLT had a genome size of 6.04 Mb and the average G+C content of 65.6 %. Compared to the other Hymenobacter spp., strain NLT is different in polar lipid profile (without aminophospholipid) and leucine arylamidase activity. Based on the data of the polyphasic analysis, it is considered that strain NLT represented a novel species of genus Hymenobacter , for which the name Hymenobacter edaphisp. nov. is proposed. The type strain is NLT (=KCTC 62521T=CCTCC AB 2018028T).

A Gram-stain-positive, strictly aerobic, motile and rod-shaped bacterium, designated strain LJ137T, was isolated from the sediment of Taihu Lake in China. A polyphasic approach was used to investigate its taxonomic position. Strain LJ137T grew optimally at pH 7.5, at 37 °C and with 2.5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain LJ137T was most closely related to the genera Ornithinibacillus and Oceanobacillus. The closest phylogenetic neighbours were Ornithinibacillus halophilus KCTC 13822T, Ornithinibacillus salinisoli LCB256T and Oceanobacillus limi KCTC 13823T, with 95.2, 96.5 and 95.6 % 16S rRNA gene sequence similarity, respectively. The peptidoglycan amino acid type was A4α (l-Lys–d-Asp). The major respiratory quinone was menaquinone-7 (MK-7). The polar lipids of strain LJ137T contained diphosphatidylglycerol, phosphatidylglycerol, three unidentified phospholipids, two aminophospholipids and one unidentified lipid. The G+C content of the genomic DNA was 40.4 mol%. The dominant cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C15 : 0. Based on the phenotypic, chemotaxonomic, phylogenetic and genome sequence characteristics of this strain, a novel species, Ornithinibacillus gellani sp. nov., is proposed. The type strain is LJ137T (=CGMCC 1.13678T=NBRC 113552T). An emended description of the genus Ornithinibacillus is presented.

Moutai-flavour Daqu is an important starter to support growth of microorganisms in the fermented process of Moutai-flavour liquor. A novel thermophilic microorganism, designated strain FBKL4.011T, was isolated from Moutai-flavour Daqu samples collected from Guotai distillery in Renhuai, Guizhou province, south-west China. The strain could grow at 45–65 °C (optimum 45 °C). Based on polyphasic analysis, strain FBKL4.011T was affiliated to the genus Thermoflavimicrobium . It formed abundant pale-yellow aerial and substrate mycelium, bearing single endospores (7.0–10.0 µm diameter) on branched long sporophores (5.0 µm diameter). The cell-wall peptidoglycan contained meso-diaminopimelic acid; ribose, glucose and mannose were the primary whole-cell sugars. The major fatty acids were iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0. The predominant menaquinone were MK-8 and MK-9. The polar phospholipids contained diphosphatidyl glycerol, phosphatidyl ethanolamine, one unidentified phospholipid and one unidentified lipid. The G+C content of the genome was 43.1 mol%. According to the 16S rRNA gene sequence analysis, strain FBKL4.011T was closely related to Thermoflavimicrobium dichotomicum JCM 9688T (95.3 % sequence similarity), and other members within the family Thermoactinomycetaceae (less than 93.0 % sequence similarity). The DNA–DNA hybridisation data showed low relatedness between strain FBKL4.011T and T. dichotomicum JCM 9688T, Laceyella sacchari KCTC 9790T, Laceyella tengchongensis YIM 10002T, Laceyella sediminis RHA1T(36.7±1.1 %, 30.0±1.2 %, 21.3±2.1 % % and 37.6±0.9 %, respectively). Based on data from the polyphasic analysis, strain FBKL4.011T is considered to represent a novel species of the genus Thermoflavimicrobium , for which the name Thermoflavimicrobium daqui sp. nov. is proposed. The type strain is FBKL4.011T(=KCTC 43036T=CICC 24504T).

Early characterization of strains designated into the genera Entomoplasma and Mesoplasma was based upon biological and chemical characteristics. With the advent of 16S rRNA gene sequence analysis as an added taxonomic character, it became clear that the two genera did not form distinct and separate monophyletic clusters. A genome-level analysis of all 17 validly published species within the family Entomoplasmataceae has recently been performed. Phylogenetic analyses, comparisons of gene content, and the lack of genus-specific genes supported that species from the two genera are intermixed and should not be taxonomically separated. This level of analysis clearly reveals the necessity to revise the taxonomy of this family by merging the two genera into one, Entomoplasma . Additionally, it was definitively determined that the strain originally designated as Acholeplasma multilocale resides in this cluster and should be formally renamed as Entomoplasma multilocale. Merging Mesoplasma and Entomoplasma yields a paraphyletic genus, but is supported by cell morphology and ecology to be distinguished from the genera Spiroplasma and Mycoplasma .