- Volume 72, Issue 3, 2022

A Gram-stain-positive bacterial strain, designated G127ATT, was isolated as soft small white colonies from the hindgut of the cockroach Archimandrita tesselata. Examination of the complete 16S rRNA sequence mapped the strain to the genus Agromyces . The type strain with the highest pairwise similarity was Agromyces marinus H23-8T (97.3%). The genome of G127ATT was sequenced by a combination of Illumina and Nanopore methods and consisted of a single circular DNA molecule with a size of 3.45 Mb. The DNA G+C content was 71.3 mol%. A phylogenomic tree based on conserved single copy housekeeping genes, placed G127ATT among the ancestral species of the genus Agromyces , and only Agromyces atrinae P27T was found to diverge earlier than G127ATT. Genome distance metrics average nucleotide identity (ANI) (76–78 %) and digital DNA–DNA hybridization (dDDH) (20.2–21.5 %) of the isolate against available genomes of several type strains of species of the genus Agromyces indicated that G127ATT represented a previously undescribed species of the genus Agromyces . Morphological, physiological and biochemical characteristics, including lipid profile, cellular fatty acids and peptidoglycan type were in accordance with usual attributes of members of the genus Agromyces . The novel isolate could be differentiated from the most closely related species by extracellular expression of acid and alkaline phosphatases, trypsin and α-chymotrypsin, and utilization of l-arabinose and salicin as sole carbon sources. On the basis of the combined genomic and phenotypic features, isolate G127ATT (=DSM 111850T=LMG 32099T) is considered to represent a novel species of the genus Agromyces , for which we propose the name Agromyces archimandritae sp. nov.

A novel bacterial strain (MSC19T) was isolated from a deep-sea sponge Cacospongia mycofijiensis collected in the Mariana Trench at a depth of 2681 m. The cells of the new isolate were Gram-stain-positive, non-motile, oxidase- and catalase-positive, rod-shaped and yellow-coloured. They could grow at 4–32 °C (optimum, 28 °C), pH 5.5–12 (optimum, pH 7.0) and with 0–12 % (w/v) NaCl (optimum, 4 %). The strain’s 16S rRNA gene sequence showed 98.41 % similarity to that of Mycetocola saprophilus CM-01T. Phylogenetic analysis further suggested that strain MSC19T represents a new species within the genus Mycetocola . The total genome of MSC19T was approximately 3 196 754 bp in size with a G+C content of 66.43 mol%. The average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values among MSC19T and other Mycetocola type strains were 70.35–75.37 % (ANIb), 83.79–84.73 % (ANIm) and 20.3–21.7 % (dDDH). The major fatty acids of MSC19T were composed of anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0, and its predominant menaquinones were MK-10 and MK-9. The polar lipids of MSC19T mainly consisted of diphosphatidylglycerol, phosphatidylglycerol and glycolipid. The diagnostic cell-wall diamino acid was lysine. Combined molecular, physiological, biochemical and chemotaxonomic analyses suggest that strain MSC19T represents a novel species of the genus Mycetocola , for which the name Mycetocola spongiae sp. nov. is proposed. The type strain is MSC19T (=MCCC 1K06265T=KCTC 49701T).

A novel species of endophytic actinobacterium, designated strain GBK3QG-3T, was isolated from surface-sterilized root of Kandelia candel collected from Beilun Estuary, Guangxi Zhuang Autonomous Region, PR China. Cells of strain GBK3QG-3T were Gram-stain-positive, aerobic, non-spore-forming and rod-shaped. Growth of GBK3QG-3T was observed at 10–40 °C (optimum, 30 °C), pH 5.0–9.0 (optimum, pH 7.0) and in the presence of 0–3 % (w/v) NaCl (optimum, 0 %). The G+C content of genomic DNA was determined to be 72.2 mol%. On the basis of 16S rRNA gene sequence analysis, strain GBK3QG-3T was most related to the type strain of Nocardioides agrisoli CCTCC AB 2017058T (97.7 % 16S rRNA gene sequence similarity), followed by Nocardioides ginkgobilobae SYP-A7303T (97.0 %) and Nocardioides aquiterrae GW-9T (96.9 %). Average nucleotide identity and digital DNA–DNA hybridization values between strain GBK3QG-3T and some strains of the genus Nocardioides with publicly available genomes were below 84.3 and 30.0 %, respectively. The cell-wall peptidoglycan contained ll-diaminopimelic acid. The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, an unknown phospholipid and an unknown lipid. The major menaquinone was identified as MK-8(H4), and the major fatty acids were identified as iso-C16 : 0, iso-C17 : 0 and anteiso-C17 : 0. Based on evidence collected from the phenotypic, genotypic and phylogenetic analyses, we propose a novel species named Nocardioides mangrovi sp. nov. The type strain is GBK3QG-3T (=CGMCC 4.7708T=JCM 34553T).

A Gram-stain-positive, non-spore-forming, yellow-pigmented, non-motile, non-flagellated, facultative anaerobic and rod-shaped bacterial strain, designated BI34T, was isolated from the gut of the larva of Protaetia brevitarsis seulensis. Strain BI34T grew at 15–40 °C (optimum, 37 °C), at pH 6.5–9.0 (optimum, pH 7.5) and in the presence of 0–7 % (w/v) NaCl (optimum, 2 %). Based on the results of 16S rRNA gene sequence analysis, strain BI34T belonged to the phylum Actinobacteria and was closely related to Cellulosimicrobium funkei NBRC 104118T (99.3 %), Cellulosimicrobium cellulans NBRC 15516T (99.1 %), Cellulosimicrobium composti BIT-GX5T (99.0 %), Cellulosimicrobium fucosivorans SE3T (99.0 %), Cellulosimicrobium marinum NBRC 110994T (98.4 %) and Cellulosimicrobium terreum DS-61T (97.0 %). The genome to genome relatedness of the average nucleotide identity (ANI) and the digital DNA–DNA hybridization (dDDH) values calculated by the Genome-to-Genome Distance Calculator between strain BI34T and its related species mentioned above were lower than the threshold of 95 and 70 % for speciation, respectively. The predominant menaquinone of strain BI34T contained MK-9(H4), and the major fatty acids were anteiso-C15 : 0, C16 : 0 and anteiso-C17 : 0. Strain BI34T had diphosphatidylglycerol and phosphatidylglycerol as major polar lipids. The whole-cell sugars were galactose, glucose and ribose, and the cell-wall peptidoglycan contained lysine, alanine, aspartic acid and glutamic acid. The DNA G+C content of strain BI34T was 73.8 mol%. The difference in physiological and biochemical characteristics and the below-threshold values of genome-to-genome relatedness indicate that strain BI34T represents a novel species in the genus Cellulosimicrobium , for which the name Cellulosimicrobium protaetiae sp. nov. is proposed. The type strain is BI34T (=KCTC 49302T=NBRC 114073T).

A novel actinomycete, strain 1_25T, was isolated from soil under a black Gobi rock sample from Shuangta, PR China, and characterized using a polyphasic taxonomic approach. The results of comparative analysis of the 16S rRNA gene sequences indicated the 1_25T represented a member of the genus Streptomyces . Chemotaxonomic data revealed that 1_25T possessed MK-9(H8) as the major menaquinone. The cell wall contained ll-diaminopimelic acid (ll-DAP) and the whole-cell sugar pattern consisted of ribose, glucose and galactose. Major fatty acid methyl esters were observed to be iso-C16 : 0 (23.6 %), and anteiso-C15 : 0 (10.4 %). The genomic DNA G+C content of 1_25T was 69 mol %. The results of phylogenetic analysis based on 16S rRNA gene sequence indicated that 1_25T had high sequence similarity with Streptomyces qinglanensis 172205T (98.1 %), Streptomyces lycii TRM 66187T (98 %), and Streptomyces griseocarneus JCM4580T (98 %). In addition to the differences in phenotypic characters, the average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values between 1_25T and closely related species were below the recommended threshold values for assigning strains to the same species. The fermentation product of 1_25T in ISP2 had an inhibitory effect on Staphylococcus aureus . On the basis of these genotypic and phenotypic characteristics, strain 1_25T (=JCM 34936T=GDMCC 4.216T) represents a novel species of the genus Streptomyces , for which the name Streptomyces gobiensis sp. nov. is proposed.

Two bacterial strains, designated BT189T and BT664T, were isolated from soil sampled in the Republic of Korea. Phylogenetic analysis based on the 16S rRNA gene sequences showed that strains BT189T and BT664T belonged to the genus Hymenobacter , family Hymenobacteraceae (order Cytophagales ). The 16S rRNA genes of the two strains shared a sequence similarity of 93.7 %. The closely related species of strain BT189T were Hymenobacter rubidus DG7BT (97.1 % 16S rRNA similarity) and Hymenobacter terrae DG7AT (96.7 %). The closest related species to strain BT664T were Hymenobacter sedentarius DG5BT (95.3 %) and Hymenobacter terrenus MIMtkLc17T (95.2 %). The genome sizes of strains BT189T and BT664T were 5 285 287 and 5 475 357 bp, respectively. The genomic DNA G+C contents of strains BT189T and BT664T were 63.2 and 59.3 mol%, respectively. The main fatty acids of strain BT189T were iso-C15 : 0, anteiso-C15 : 0 and summed feature 3 (C16 : 1  ω6c/C16 : 1  ω7c), and those of strain BT664T were iso-C15 : 0, C16 : 1  ω5c and summed feature 3 (C16 : 1  ω6c/C16 : 1  ω7c). The main polar lipid in both strains was phosphatidylethanolamine and the predominant respiratory quinone was MK-7, supporting the affiliation of these strains with the genus Hymenobacter . Based on the results of biochemical, chemotaxonomic and phylogenetic analyses, two novel species, Hymenobacter armeniacus BT189T (=KCTC 72341T=NBRC 114843T) and Hymenobacter montanus BT664T (KACC 21967T=NBRC 114856T), are proposed.

The genus Prevotella plays an important role in polysaccharide degradation and fermentation in the rumen. To further understand the function of the phylogenetically diverse genus Prevotella , it is necessary to explore the individual characteristics at the species level. In this study, Gram-negative anaerobic bacterial strains isolated from the rumen of Holstein cows were identified. Strain R5019T was classified within the genus Prevotella based on 16S rRNA gene sequence-based phylogenetic analysis. The values of 16S rRNA gene sequence similarity, average nucleotide identity and digital DNA–DNA hybridization between strain R5019T and its phylogenetically nearest species Prevotella multisaccharivorax PPPA20T were 89.8, 82.6, and 29.3 %, respectively. The genome size of R5019T was estimated to be ca. 4.19 Mb with a genomic G+C content of 49.5 mol%. The major cellular fatty acids and menaquinones were C15 : 0 anteiso and C17 : 0 anteiso and MK-11 and MK-12, respectively. Succinate, lactate, malate, acetate and formate were produced as the fermentation end products using glucose. Based on phylogenetic, physiological, biochemical and genomic differences between 11 strains and other phylogenetically related Prevotella species, a novel species, Prevotella lacticifex sp. nov., is proposed within the genus Prevotella . The type strain is R5019T (=JCM 34664T=DSM 112675T).

Gram-stain-negative, strictly aerobic, red-pink-coloured, rod-shaped and non-motile bacterial strains PAMC 29290, PAMC 29294T and PAMC 29296 were isolated from marine surface sediment sampled in the East Siberian Sea and strains PAMC 26553 and PAMC 26554T were obtained from an Antarctic lichen. Strains PAMC 29290, PAMC 29294T and PAMC 29296 were closely related to Hymenobacter artigasi (98.8 % 16S rRNA gene similarity), Hymenobacter antarcticus (97.3 %) and Hymenobacter glaciei (96.9 %), and PAMC 26553 and PAMC 26554T showed high similarity to Hymenobacter ginsengisoli (97.0 %), Hymenobacter rivuli (96.1 %) and Hymenobacter setariae (95.9 %). Genomic relatedness analyses showed that strains PAMC 29290, PAMC 29294T and PAMC 29296 could be distinguished from H. artigasi by average nucleotide identity (ANI; 93.1–93.2 %) and digital DNA–DNA hybridization (dDDH; 50.3–51.0 %) values. Strains PAMC 26553 and PAMC 26554T could be clearly distinguished from H. ginsengisoli with ANI values <79.8 % and dDDH values <23.3 %. The major fatty acids of strains PAMC 29290, PAMC 29294T and PAMC 29296 were C15 : 0 iso (21.0–26.0 %), summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c; 17.4–18.2 %), C15 : 0 anteiso (12.7–19.1 %) and summed feature 4 (C17 : 1 iso I and/or anteiso B; 8.6–16.1 %) and those of strains PAMC 26553 and PAMC 26554T were summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c; 20.7–22.2 %), C15 : 0 anteiso (17.5–19.7 %) and summed feature 4 (C17 : 1 iso I and/or anteiso B; 15.5–18.1 %). The major respiratory quinone was MK-7. The genomic DNA G+C contents were 60.6–60.8 mol%. The polar lipids of PAMC 29294T were found to consist of phosphatidylethanolamine, four unidentified aminolipids, an unidentified aminophospholipid and five unidentified lipids; those of PAMC 26554T were phosphatidylethanolamine, three unidentified aminolipids, four unidentified aminophospholipid and two unidentified lipids. The distinct phylogenetic position and some physiological characteristics distinguished the novel strains from closely related type strains in the genus Hymenobacter . Thus, two novel species are proposed, with the names Hymenobacter siberiensis sp. nov. (type strain, PAMC 29294T=KCTC 82466T=JCM 34574T) and Hymenobacter psoromatis sp. nov. (type strain, PAMC 26554T=KCTC 82464T=JCM 34572T), respectively.

A Gram-stain-negative, non-motile, aerobic, yellow, convex, rod-shaped mesophilic bacterial strain, designated strain D33T, was isolated from rhizosphere soil of ancient mulberry in Dezhou city, Shandong province, PR China. The strain grew at 8–37 °C (optimum, 30 °C), pH 4–9 (optimum, pH 7) and growth occurred at 0.5–5.5 % (w/v) NaCl (optimally at 1 %). The results of the phylogenetic analyses of 16S rRNA gene and whole genome sequences indicated that D33T was closely related to members of the genus Flavobacterium and had the highest 16S rRNA gene sequence similarity with ‘Flavobacterium agri’ KACC 19300 (95.4 %), Flavobacterium ichthyis NST-5T (94.6 %), Flavobacterium ahnfeltiae KCTC 32467T (93.6 %) and Flavobacterium longum JCM 19141T (93.6 %). The genome size of D33T was 3.8 Mb and the DNA G+C content was 48.0 mol%. The average nucleotide identity (ANI), digital DNA–DNA hybridization (dDDH) and average amino acid identity (AAI) values among D33T and reference strains were lower than the threshold values for species delineation. The only respiratory quinone of D33T was menaquinone 6 (MK-6). The predominant fatty acids (>5 %) were C15:0, C16 : 0, C18 : 0, iso-C15:0, iso–C17 : 0 3-OH, anteiso-C15 : 0 and summed feature 9 . The polar lipid profile contained phosphatidylethanolamine, two unidentified aminophospholipids, three unidentified aminolipids and two unidentified lipids. Combined data from phenotypic, phylogenetic and chemotaxonomic studies indicated that D33T is a representative of a novel species of the genus Flavobacterium, for which the name Flavobacterium selenitireducens sp. nov. is proposed. The type strain is D33T (=GDMCC 1.1946T=KACC 22131T).

A Gram-negative, non-motile, strictly aerobic and rod- or filamentous-shaped strain, CJU-R4T, was isolated from a flower of royal azalea (Rhododendron schlippenbachii) collected in the Republic of Korea. Strain CJU-R4T was catalase-positive and oxidase-negative, and grew at 15–33 °C (optimum, 28–20 °C), at pH 5.0–8.0 (optimum, pH 7.0–8.0), and in the presence of 0–1 % NaCl (w/v; optimum, 0 %). Strain CJU-R4T had the highest 16S rRNA gene sequence similarity to Spirosoma oryzae RHs22T (96.6 %), revealing less than 93 % sequence similarity to other type strains. Phylogenetic and phylogenomic analysis also revealed strain CJU-R4T formed a robust cluster with S. oryzae RHs22T. The major fatty acids were summed feature 3 (comprising C16 : 1  ω7c and/or C16 : 1  ω6c; 33.0 %), C16 : 1  ω5c (22.1 %), iso-C15 : 0 (12.6 %) and C16 : 0 (10.7 %). The polar lipids were composed of phosphatidylethanolamine, three unidentified aminophospholipids, one unidentified phospholipid and four unidentified lipids. Menaquinone-7 was detected as the sole respiratory quinone. The genomic DNA G+C content was 55.2 mol%. The average nucleotide identity and digital DNA–DNA hybridization values between strain CJU-R4T and Spirosoma oryzae DSM 28354T were 81.5 and 23.9 %, respectively. Based on the results of the phenotypic and genotypic analyses, strain CJU-R4T is considered to represent a novel species of the genus Spirosoma , for which the name Spirosoma rhododendri sp. nov. is proposed. The type strain is CJU-R4T (=KACC 21264T=NBRC 114513T).

Three aerobic, Gram-stain-negative, non-motile, rod-shaped bacteria, designated as strains SHINM13T, GENT5T and GENT11 were isolated from surface river water (Saitama Prefecture, Japan). SHINM13T and GENT11 were positive for catalase, whereas GENT5T was negative. Phylogenetic analyses based on the 16S rRNA gene (1341 bp) or 40 marker gene (34,513 bp) sequences revealed that the strains formed distinct phylogenetic lineages within the genus Flavobacterium . The three strains shared 99.3–99.6 % 16S rRNA gene sequence similarity among each other. The average nucleotide identity by orthology (OrthoANI) and digital DNA–DNA hybridization (dDDH) values between strains SHINM13T and GENT11 were 96.56 and 82.1 %, respectively, and those between SHINM13T and GENT5T were 83.46 % and 52.9 %, respectively. The major cellular fatty acids were C15 : 1ω6c, iso-C15 : 0, iso-C15 : 1G, anteiso-C15 : 0 and iso-C15 : 0 3-OH. The major polar lipid was phosphatidylethanolamine. SHINM13T and GENT5T contained menaquinone-6 (MK-6) as the predominant respiratory quinone, and their DNA G+C contents were 34.4 and 35.1 mol%, respectively. Genome sequencing of the three isolates revealed a genome size of 2.26–2.40 Mbp. Furthermore, all three isolates converted dissolved organic nitrogen to ammonium during cell growth. On the basis of the results of phenotypic and phylogenetic analyses, strains SHINM13T and GENT11 and GENT5T represent two distinct novel species in the genus Flavobacterium , for which the names Flavobacterium ammonificans sp. nov. (type strain SHINM13T =JCM 34684T =NCIMB 15379T) and Flavobacterium ammoniigenes sp. nov. (type strain GENT5T =JCM 32249T=NCIMB 15380T) are proposed.

Two novel, designated strains 29W222T and 2943T, were isolated from the marine sediment from Aoshan Bay, Jimo, PR China. Growth was observed at pH 6.0–8.5 (optimum, pH 7.5) for strain 29W222T, and pH 5.5–8.5 (pH 7.0) for strain 2943T. Both strains displayed growth in 0.5–6 % NaCl with an optimum at 1 % for 29W222T; 0.5 % for 2943T. Both strains grew optimally at 33 °C. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that 29W222T and 2943T represented members of the genus Fulvivirga and strain 29W222T was most closely related to Fulvivirga kasyanovii KMM 6220T (97.9 % sequence similarity) and Fulvivirga imtechensis AK7T (95.0 %), and 2943T to Fulvivirga imtechensis AK7T (95.7 %) and Fulvivirga kasyanovii KMM 6220T (94.8 %). The genomic DNA G+C contents of 29W222T and 2943T were 39.9 and 37.7 mol%, respectively. The results of chemotaxonomic analysis indicated that the sole respiratory quinone was menaquinone 7 (MK-7), and the major fatty acid was iso-C15 : 0 for both strains. Average nucleotide identity and average amino acid identity values between strain 29W222T and Fulvivirga kasyanovii KMM 6220T were 78.9 and 83.6 %, respectively; the corresponding values between 2943T and Fulvivirga imtechensis AK7T were 69.8 and 63.6 %, respectively. Therefore, strains 29W222T and 2943T represent to two novel species of the genus Fulvivirga , for which the names Fulvivirga marina sp. nov. (29W222T=KCTC 62848T=MCCC 1K05194T) and Fulvivirga sediminis sp. nov. (2943T=KCTC 62847T= MCCC 1K05144T) are proposed, respectively.

Two Gram-positive, endospore-forming, rod-shaped bacterial strains designated MWE-103T and DLE-14T were isolated from plant roots. The 16S rRNA gene sequence analysis indicated that strain MWE-103T was closely related to Paenibaillus sacheonensis SY01T with a sequence similarity of 97.82 %, and strain DLE-14T to Paenibacillus rhizoryzae IZS3-5T with 99.09 % similarity. The orthologous average nucleotide identity and digital DNA–DNA hybridization values using whole genome data indicated that strains MWE-103T and DLE-14T could be readily distinguished from the mostly related species. Both strains grew at mesophilic temperature ranges, and grew best at pH 6 and in the absence of NaCl. The major fatty acid in both strains was anteiso-C15 : 0, but their relative proportions differed. The predominant quinone of both strains was menaquinone 7, the cell-wall diamino acid was meso-diaminopimelic acid, and the diagnostic polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol, which were consistent with those of related species. Amylase and cellulase activities were positive for both strains. Strain DLE-14T exhibited the potential for lignin degradation. The DNA G+C contents of strain MWE-103T and DLE-14T were 60.9 and 50.8 mol% respectively. The genomes of the two strains revealed potential plant-growth-promoting characteristics such as nitrogen fixation, siderophore production and phosphate solubilization. Based on phylogenetic and phenotypic evidence, strains MWE-103T and DLE-14T should each be recognized as a novel species of Paenibacillus , for which the names Paenibacillus artemisiicola sp. nov. (type strain: MWE-103T=KCTC 43287T=JCM 34503T) and Paenibacillus lignilyticus sp. nov. (type strain: DLE-14T=KCTC 43288T=JCM 34504T) are proposed.

Non-human primates harbour diverse microbiomes in their guts. As a part of the China Microbiome Initiatives, we cultivated and characterized the gut microbiome of cynomolgus monkeys (Macaca fascicularis). In this report, we communicate the characterization and taxonomy of eight bacterial strains that were obtained from faecal samples of captive cynomolgus monkeys. The results revealed that they represented eight novel bacterial species. The proposed names of the eight novel species are Alkaliphilus flagellatus (type strain MSJ-5T=CGMCC 1.45007T=KCTC 15974T), Butyricicoccus intestinisimiae MSJd-7T (MSJd-7T=CGMCC 1.45013T=KCTC 25112T), Clostridium mobile (MSJ-11T=CGMCC 1.45009T=KCTC 25065T), Clostridium simiarum (MSJ-4T=CGMCC 1.45006T=KCTC 15975T), Dysosmobacter acutus (MSJ-2T=CGMCC 1.32896T=KCTC 15976T), Paenibacillus brevis MSJ-6T (MSJ-6T=CGMCC 1.45008T=KCTC 15973T), Peptoniphilus ovalis (MSJ-1T=CGMCC 1.31770T=KCTC 15977T) and Tissierella simiarum (MSJ-40T=CGMCC 1.45012T=KCTC 25071T).

A bacterial strain designated BG109T was isolated from bamboo grove soil, and subjected to polyphasic taxonomic characterization. BG109T is an aerobic, non-motile, Gram-stain-positive and endospore-forming bacterium. BG109T showed growth at 10–40 °C (optimum, 37 °C), at pH 4–10 (optimum, 8), and in the presence of 0–7 % NaCl concentration (optimum, 0–1 %). The predominant menaquinone of BG109T was MK-7, and the cell wall peptidoglycan contained major amounts of meso–diaminopimelic acid as the diagnostic diamino acid. The diagnostic polar lipids were diphosphatidylglycerol and phosphatidylglycerol, and unidentified phospholipids and glycolipids were also present. The major fatty acids were anteiso-C15 : 0, iso-C15 : 0, iso-C14 : 0, iso-C16 : 0 and C16 : 0. The chemotaxonomic properties of BG109T were generally consistent with those of members of the genus Metabacillus . BG109T shared highest 16S rRNA gene sequence similarities with ‘Metabacillus elymi’ KUDC1714 (99.26 %), Metabacillus sediminilitoris DSL-17T (98.17 %), Metabacillus litoralis SW-211T (98.16 %) and Metabacillus crassostreae JSM 100118T (97.13 %), all of which were well below the suggested cutoff level for species distinction. The genome level relatedness also confirmed the separation of BG109T from other species of the genus Metabacillus . Thus, it is evident that BG109T merits recognition as representing a novel species of the genus Metabacillus , for which the name Metabacillus bambusae sp. nov. is proposed. The type strain is BG109T (=KCTC 43190T=JCM 34515T).

The genera   Convivina  ,   Fructobacillus  ,   Leuconostoc  ,   Oenococcus   and   Weissella  , which formed the family   Leuconostocaceae  , have recently been merged within the family   Lactobacillaceae  . Using genome sequences for 47 of the 52 named species from these genera, we report here comprehensive phylogenomic and comparative analyses on protein sequences from these species using multiple approaches. In a phylogenomic tree based on concatenated sequences of 498 core proteins from these five genera, and in a 16S rRNA gene tree, members of the genera   Fructobacillus  ,   Leuconostoc   and   Oenococcus   formed distinct strongly supported clades. In contrast,   Weissella   species grouped into two distinct unrelated clades designated as the ‘  Weissella   main clade’ and ‘  Weissella   clade 2’. The presence of these clades is also seen in a matrix of pairwise average amino acid identity based on core protein sequences. In parallel, comparative genomic studies on protein sequences from   Leuconostocaceae   genomes have identified 46 conserved signature indels (CSIs) in diverse proteins that are unique characteristics of the different observed species clades. Of these identified CSIs, five, five and 13 CSIs are uniquely present in members of the genera   Fructobacillus  ,   Leuconostoc   and   Oenococcus  , respectively. We also report here six and five CSIs that are exclusively present in the species from the   Weissella   main clade and   Weissella   clade 2, respectively, providing independent evidence supporting their distinctness from each other. The remaining 12 identified CSIs are commonly shared by some or all of the species from the genera   Convivina  ,   Fructobacillus   and   Leuconostoc  , clarifying their interrelationships. The identified CSIs provide novel and reliable means for the identification/circumscription of members of the genera   Fructobacillus  ,   Leuconostoc   and   Oenococcus   as well as the two   Weissella   species clades in molecular terms. Based on the strong phylogenetic and molecular evidence presented here, we propose that the genus   Weissella   be limited to only the species from the   Weissell  a main clade, whereas the species forming   Weissella   clade 2 should be transferred to a new genus Periweissella gen. nov.

Two acidophilic strains, designated as ALEF1T and S30H14T, were isolated from acid mine drainage sediment. Cells of both strains were Gram-stain-positive, aerobic, endospore-forming rods. Strains ALEF1T and S30H14T were acidophilic and mesophilic, the former grew at 20–40 °C (optimum, 30 °C) and pH 2.5–4.5 (optimum, pH 3.5), while the latter grew at 20–45 °C (optimum, 30 °C) and pH 2.0–5.5 (optimum, pH 4.5). The 16S rRNA gene-based sequence analysis revealed that strains ALEF1T and S30H14T belonged to the genus Alicyclobacillus , and were phylogenetically close to Alicyclobacillus ferrooxydans TC-34T with 97.1 and 97.4% similarity, respectively. The similarity between the two novel strains was 98.6 %. The average nucleotide identity value between the genome sequences of ALEF1T and S30H14T was 79.5 %, and that between each of the two isolates and A. ferrooxydans TC-34T were 72.0 and 74.3 %. In addition, the digital DNA–DNA hybridization value between ALEF1T and S30H14T was 24.9 %, between strain ALEF1T and A. ferrooxydans TC-34T was 21.7 %, and between S30H14T and A. ferrooxydans TC-34T was 26.3 %, far below the interspecies threshold. Both strains could utilize diverse carbon sources for heterotrophic growth; strain ALEF1T could utilize ferrous iron as the energy source for autotrophic growth. Menaquinone 7 was the only quinone detected in either strain. Both strains contained anteiso-C15 : 0 and anteiso-C17 : 0, while ω-alicyclic fatty acids were not detected. Based on their phylogenetic positions, as well as phenotypic and genomic data, it is considered that strains ALEF1T and S30H14T represent two novel species within the genus Alicyclobacillus , for which the names Alicyclobacillus curvatus sp. nov. (type strain ALEF1T=CGMCC 1.17055T=KCTC 43124T) and Alicyclobacillus mengziensis sp. nov. (S30H14T=CGMCC 1.17050T=KCTC 43125T) are proposed.