- Volume 72, Issue 2, 2022

A polyphasic taxonomic study was carried out on an actinobacterial strain (AN110305T) isolated from soil sampled in the Republic of Korea. Cells of the strain were Gram-stain-positive, aerobic, non-motile and rod-shaped. Comparative 16S rRNA gene sequence studies showed a clear affiliation of strain AN110305T with Actinomycetia , with highest pairwise sequence similarities to Goodfellowiella coeruleoviolacea DSM 43935T (97.6%), Umezawaea tangerina MK27-91F2T (97.0%), Kutzneria chonburiensis NBRC 110610T (96.9%), Kutzneria buriramensis A-T 1846T (96.8%), Umezawaea endophytica YIM 2047XT (96.8%), Kutzneria albida NRRL B-24060T (96.7%) and Saccharothrix coeruleofusca NRRL B-16115T (96.6%). Cells of strain AN110305T formed pale-yellow colonies on Reasoner's 2A agar. MK-9 (H4) (68%) and MK-10 (H4) (32%) were the predominant menaquinones. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethyl ethanolamine, hydroxy-phosphatidylethanolamine, an unidentified aminolipid and an unidentified aminophospholipid were major polar lipids. Iso-C16:0 (24.5%), anteiso-C15:0 (19.3%), anteiso-C17:0 (15.7%) and iso-C15:0 (15.2%) were the major fatty acids and meso-diaminopimelic acid was the pepdidoglycan. The cell-wall sugars were composed of galactose, glucose, mannose and ribose. The genomic DNA G+C content was 70.7 mol%. Based on genotypic and phenotypic data, strain AN110305T could be distinguished from all genera within the family Pseudonocardiaceae and represents a novel genus and species named Solihabitans fulvus gen. nov., sp nov. The type strain is AN110305T (=KCTC 39307T =DSM 103572T).

Four novel bacterial strains (zg-ZUI122T/zg-ZUI10 and zg-ZUI227T/zg-ZUI100) were isolated from the intestinal contents of Marmota himalayana and characterized using a polyphasic approach. Cells were Gram-stain- and catalase-positive, urease- and oxidase-negative. Strains grew optimally at 28–30 °C, pH 7.0, with 0.5 % NaCl (w/v). A comparative analysis of 16S rRNA gene sequences revealed that strain pairs zg-ZUI122T/zg-ZUI10 and zg-ZUI227T/zg-ZUI100 belonged to the genus Arthrobacter and were most closely related to Arthrobacter citreus DSM 20133T, with similarities of 99.6 and 99.5 %, respectively. This was further confirmed by phylogenetic analyses based on the 16S rRNA gene and genome sequences. The digital DNA–DNA hybridization and average nucleotide identity values between the two new type strains (zg-ZUI122T and zg-ZUI227T) and other species in the genus Arthrobacter were 20.0–24.4/77.2–83.4% and 19.9–25.1/77.1–83.4%, all below the thresholds. The major cellular fatty acids detected in the two novel species included iso-C15 : 0 and anteiso-C15 : 0; the predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol. MK-8(H2) (77.3%) was the predominant respiratory quinone detected in strain zg-ZUI122T, while MK-8(H2) (53.7%) and MK-9(H2) (46.3%) were detected in strain zg-ZUI227T. The shared cell-wall amino acids detected in the two novel species were alanine, glutamic acid and lysine; the shared whole cell wall sugars consisted of galactose, mannose and ribose. All these analyses concluded that these four strains represent two different novel species in the genus Arthrobacter , for which the names Arthrobacter sunyaminii sp. nov. (zg-ZUI122T = GDMCC 1.2502T = KCTC 49677T) and Arthrobacter jiangjiafuii sp. nov. (zg-ZUI227T = GDMCC 1.2500T = KCTC 49676T) are proposed.

A novel micro-organism designated AS10T was isolated from dry salt collected from Aveiro saltern in the north of Portugal. Cells were Gram-stain-positive, non-motile, non-endospore-forming, rod-shaped and aerobic. Strain AS10T was characterized by long filaments of rod-shaped cells, presenting also coccoid cellular forms at the end of the filaments, unveiling some pleomorphism. Rod-shaped cells varied from 0.3 to 0.6 µm wide and from 0.6 to 2 µm long. Growth of AS10T occurred at 15–40 °C (optimum, 20–30 °C), 0–10% (w/v) NaCl (optimum, 2%) and pH 4.5–11.0 (optimum, pH 8.0–11.0). The peptidoglycan type was A1ϒ-type with 3-hydroxy-diaminopimelic acid. The major fatty acids were C16:0, iso-C14:0, C17:0 and C14:0. The major respiratory quinone was MK-9(H4). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AS10T was similar to actinobacterial members of the class   Nitriliruptoria  , with   Nitriliruptor alkaliphilus   ANL-iso2T being the closest relative the species with a sequence pairwise similarity of 91.21%. Average nucleotide identity, average amino acid identity and in silico DNA–DNA hybridization values between strain AS10T and   N. alkaliphilus   ANL-iso2T were 71.34, 53.57 and 18.90%, respectively. The genome DNA G+C content of AS10T was 71.8 mol%. Based on genomic, phylogenetic, phenotypic and chemotaxonomic studies, we describe a new species of a novel genus represented by strain AS10T (=LMG 31937T=CECT 30148T) for which we propose the name Salsipaludibacter albus gen. nov., sp. nov. We also propose that this organism represents a new family named Salsipaludibacteraceae fam. nov. of a novel order named Salsipaludibacterales ord. nov.

Two new marine actinobacteria, designated as J2-1T and J2-2T, were isolated from a coral, Favites pentagona, collected from Rayong Province, Thailand. The taxonomic positions of the two strains were identified based on polyphasic taxonomy. Based on morphological characteristics and chemotaxonomy, strains J2-1T and J2-2T were identified as members of the genus Streptomyces and Kineosporia , respectively. Strains J2-1T and J2-2T showed the highest 16S rRNA gene sequence similarity to Streptomyces broussonetiae T44T (98.62 %) and Kineosporia babensis VN05A0415T (98.08 %), respectively. Strain J2-1T had chemotaxonomic properties resembling members of the genus Streptomyces . ll-Diaminopimelic acid, glucose and ribose were detected in the whole-cell hydrolysate. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositolmannoside, unidentified aminolipid and five unidentified phospholipids were detected as the polar lipids. The major cellular fatty acids were C16 : 0 iso, C15 : 0 anteiso, C15 : 0 iso, C16 : 0, C17 : 0 anteiso, C14 : 0 iso and C17 : 0 iso. Strain J2-2T a showed similar cell composition to members of the genus Kineosporia . Both isomers of ll- and meso-diaminopimelic acid were detected in the peptidoglycan. Arabinose, galactose, madurose and xylose were observed in the whole-cell hydrolysate. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, phosphatidylcholine, an unidentified phospholipid and an unidentified glycolipid. The major cellular fatty acids were C16 : 0, C18 : 1 ω9c, C18 : 0 10-methyl, tuberculostearic acid, C18 : 0 and C17 : 0. Both strains could be distinguished from their closely related type strains according to their phenotypic characteristics. Comparative genome analysis indicated the delineation of two novel species based on digital DNA–DNA hybridization and average nucleotide identity values, which were below 70 and 95 %, respectively. The names proposed are Streptomyces corallincola sp. nov. (J2-1T=TBRC 13503T=NBRC 115066T) and Kineosporia corallincola sp. nov. (J2-2T=TBRC 13504T=NBRC 114885T).

A halophilic archaeal strain, designated C46T, was isolated from an inland salt lake in Qinghai Province, PR China. Results of phylogenetic analysis based on 16S rRNA gene sequences indicated that strain C46T belongs to the genus Halobaculum , and the closest phylogenetic relative is Halobaculum gomorrense DSM 9297T with 97.7 % similarity. Despite this, strain C46T was more related to Halobaculum saliterrae WSA2T than other members of the genus Halobaculum based on genome comparison and analysis, and the average nucleotide identity, in silico DNA–DNA hybridization, amino acid identity and percentage of conserved protein values between the two strains were 89.1, 53.3, 89.2 and 75.6 %, respectively, which are lower than the cutoff values proposed for species delimitation. The physiological, biochemical, genetic and genomic characteristics of strain C46T were different from those of its closest phylogenetic neighbours, which indicated that this strain represents a novel species of the genus Halobaculum , for which the name Halobaculum rubrum sp. nov. is proposed. The type strain is C46T (=CGMCC 1.13737T=JCM 32959T).

A moderately halophilic bacterium, designated strain KX20305T, was isolated from sediment collected from a cold seep field in the South China Sea. Cells of strain KX20305T were Gram-stain-negative, rod-shaped, non-motile, facultatively anaerobic, oxidase- and catalase-positive, and grew optimally at 25–30 °C, pH 6.0–8.0 and with 3–6 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KX20305T grouped with members of the genus Aequorivita , including Aequorivita aquimaris D-24T (98.3 % sequence similarity), Aequorivita vladivostokensis KMM 3516T (98.1 %) and Aequorivita echinoideorum CC-CZW007T (97.5 %). Genome sequencing of strain KX20305T revealed a genome size of 3.35 Mb and a DNA G+C content of 38.71 mol%. Genomic average nucleotide identity (orthoANI) values of strain KX20305T with A. aquimaris D-24T, A. vladivostokensis KMM 3516T and A. echinoideorum JCM 30378T were 83.8, 81.7 and 75.4 %, respectively, while in silico DNA–DNA hybridization (GGDC) values for strain KX20305T with these strains were 27.2, 25.0 and 19.6 %, respectively. The major fatty acids of strain KX20305T were iso-C15 : 0, iso-C17 : 0 3-OH and 10-methyl C16 : 0/iso-C17 : 1  ω9c. The predominant respiratory quinone was menaquinone-6 (MK-6). The polar lipids mainly comprised phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids. Based on comparative analysis of phylogenetic, phylogenomic, phenotypic and chemotaxonomic characteristics, strain KX20305T represents a novel species of the genus Aequorivita , for which the name Aequorivita iocasae sp. nov. is proposed. The type strain is KX20305T (=KCTC 82699T=MCCC 1K06238T=JCM 34635T).

Two novel Gram-stain-negative, aerobic, rod-shaped, carotenoid-pigmented and non-flagellated bacteria, designated BC31-1-A7T and BC31-3-A3T, were isolated from polyethylene-terephthalate-degrading bacterial consortia enriched from deep-sea sediment collected in the Pacific Ocean. Optimal growth of both strains was observed at 28–32 °C, at pH 7.5 and in the presence of 3–4% (w/v) NaCl. The 16S rRNA gene sequence analysis revealed that strains BC31-1-A7T and BC31-3-A3T were closely related to Muricauda aquimarina JCM 11811T, Muricauda lutimaris KCTC 22173T, Muricauda ruestringensis DSM 13258T, Muricauda zhangzhouensis DSM 25030T, Muricauda oceani JCM 33902T and Muricauda oceanensis KCTC 72200T with 96.8–98.9% sequence similarity. The 16S rRNA gene sequence similarity between strains BC31-1-A7T and BC31-3-A3T was 97.5%. The genomic G+C contents of strains BC31-1-A7T and BC31-3-A3T were 42.1 and 41.6 mol%, respectively. The average nucleotide identity and digital DNA–DNA hybridization values between strain BC31-3-A3T, strain BC31-1-A7T and their six closely related type strains were 77.6–84.3% and 20.5–27.9%, respectively. Menaquinone-6 was detected as the major isoprenoid quinone in all strains. Their major fatty acids were iso-C15:0, iso-C15:1 G and iso-C17:0 3-OH. The major polar lipids of strains BC31-1-A7T and BC31-3-A3T were identified as one phosphatidylethanolamine, some unidentified polar lipids and one aminolipid. Based on their distinct taxonomic characteristics, strains BC31-1-A7T and BC31-3-A3T represent two novel species in the genus Muricauda . The names proposed to accommodate these two strains are Muricauda aurea sp. nov. and Muricauda profundi sp. nov., and the type strains are BC31-1-A7T (=MCCC M23246T=KCTC 82569T) and BC31-3-A3T (=MCCC M23216T=KCTC 82302T), respectively.

A Gram-stain-negative, facultatively anaerobic, motile by gliding, rod-shaped, oxidase- and catalase-positive bacterial strain, designated BB8T, was isolated from the stems of a Korean soybean cultivar (Glycine max L. cv. Gwangan). The strain produced a yellow pigment on tryptic soy agar. Growth of strain BB8T occurred at pH 5.0–8.0 (optimum, pH 7.0), at 10–35 °C (optimum, 25–30 °C) and in the presence of 0–1 % (w/v) NaCl (optimum, 0.5%). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BB8T formed a lineage within the genus Flavobacterium and was most closely related to Flavobacterium artemisiae SYP-B1015T (96.9 % 16S rRNA gene sequence similarity) and Flavobacterium ustbae T13T (96.8%). The complete genome sequence of strain BB8T was 5 513 159 bp long with a G+C content of 34.1 mol%. The major fatty acids (>10 %) of strain BB8T were iso-C15 : 0 (21 %), summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c, 20.3%) and iso-C16 : 0 3-OH (13.7%). The predominant polar lipids were phosphatidylethanolamine and unidentified aminolipids, and the major respiratory quinone was menaquinone-6. Based on these phenotypic, genotypic and chemotaxonomic characteristics, strain BB8T is considered to represent a novel species of the genus Flavobacterium , for which the name Flavobacterium endoglycinae sp. nov. is proposed. The type strain is BB8T (=KCTC 82167T=CCTCC AB 2020070T).

A novel Gram-stain-negative, aerobic, oxidase-positive, catalase-positive, non-motile, short rod-shaped, red-pigmented strain, designated as SYSU D00434T, was isolated from a dry sandy soil sample collected from the Gurbantunggut desert in Xinjiang, north-west PR China. Strain SYSU D00434T was found to grow at 4–37 °C (optimum, 28–30 °C), pH 6.0–8.0 (optimum, pH 7.0) and with 0–1.5 % (w/v) NaCl (optimum, 0–0.5 %). The predominant respiratory quinone was MK-7 and the major fatty acids (>10 %) were C16 : 1  ω5c, iso-C15 : 0, summed feature 3 (C16 : 1  ω6c and/or C16 : 1  ω7c) and summed feature 4 (anteiso-C17 : 1 B and/or iso-C17 : 1 I). The polar lipids consisted of phosphatidylethanolamine, two unidentified polar lipids, two unidentified aminolipids, two unidentified phospholipids and two unidentified glycolipids. The genomic DNA G+C content of strain SYSU D00434T was 50.6 mol%. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain SYSU D00434T belonged to the family Hymenobacteraceae , and shared a sequence similarity of less than 94.6 % to all validly named taxa. Based on the phenotypic, phylogenetic and chemotaxonomic properties, strain D00434T is proposed to represent a new species of a new genus, named Sabulibacter ruber gen. nov., sp. nov., within the family Hymenobacteraceae . The type strain is SYSU D00434T (=CGMCC 1.18624T=KCTC 82276T=MCCC 1K04975T).

Three novel strains of Gram-stain-negative, obligately anaerobic, spore-forming straight or slightly curved rods with pointed ends occurring singly or in pairs were isolated from the faeces of healthy human children. The strains were characterized by mesophilic fermentative metabolism and production of acetate, ethanol and H2 as the end metabolic products. Strains ASD3451 and ASD5720T were motile, fermented lactose and raffinose, and weakly fermented maltose. Strain ASD4241T was non-motile and did not ferment the carbohydrates listed above but fermented starch. Strains ASD3451 and ASD5720T shared average nucleotide identity higher than 98.5 % with each other, while ASD4241T had only 88.5-89 % identity to them. Based on phylogenetic and chemotaxonomic analyses, we propose Diplocloster agilis gen. nov., sp. nov. (ASD5720T=JCM 34353T=VKM B-3497T) and Diplocloster modestus sp. nov. (ASD4241T=JCM 34351T=VKM B-3498T) within the family Lachnospiraceae .

Five Gram-stain-positive strains (M1-10T, M1-13, M1-21T, M2-14T and S1-1T) were isolated from paper mulberry (Broussonetia papyrifera) in Taiwan. Cells were rod-shaped, non-motile, non-haemolytic, asporogenous, facultatively anaerobic, heterofermentative, and did not exhibit catalase and oxidase activities. Phylogenetic analysis of the 16S rRNA gene sequences revealed that these novel strains belonged to the genus Fructobacillus . On the basis of 16S rRNA gene sequence similarities, the type strains of Fructobacillus fructosus and Fructobacillus durionis were the closest neighbours to strains M1-10T, M1-13, M1-21T, M2-14T and S1-1T. Sequence analyses of concatenated two partial housekeeping genes, the RNA polymerase beta subunit (rpoC) and recombinase A (recA) also indicated that the novel strains belonged to the genus Fructobacillus . The 16S rRNA and concatenated rpoC and recA gene sequence similarities between strains M1-10T and M1-13 were 100 %, respectively. The average nucleotide identity values of M1-10T, M1-21T, M2-14T and S1-1T with F. fructosus and F. durionis were 75.1–78.9% and 76.5–77.5 %, respectively. The digital DNA–DNA hybridization values were 19.7–21.5% and 19.6–20.4 %, respectively. Phenotypic and genotypic test results demonstrated that these strains represent four novel species of the genus Fructobacillus , for which the names Fructobacillus papyriferae sp. nov., Fructobacillus papyrifericola sp. nov., Fructobacillus broussonetiae sp. nov. and Fructobacillus parabroussonetiae sp. nov. are proposed with the type strains M1-10T (=BCRC 81237T=NBRC 114433T), M1-21T (=BCRC 81239T=NBRC 114435T), M2-14T (=BCRC 81240T=NBRC 114436T) and S1-1T (=BCRC 81241T=NBRC 114437T), respectively.

Four Gram-stain-positive bacterial strains were isolated from the gut of honeybee (Apis mellifera) in China. These strains were characterized using a polyphasic taxonomic approach. The data demonstrated that three of the four strains represented two novel species of the genus Lactobacillus , strains F306-1T and F551-2T were designated as the type strains. Results of 16S rRNA gene sequence analysis indicated that strains F306-1T, F447 and F551-2T were phylogenetically related to the type strains of Lactobacillus kimbladii and Lactobacillus kullabergensis , having 99.1–99.7 % 16S rRNA gene sequence (about 1400 bp) similarities. The phylogenetic tree based on concatenated pheS, rpoA, gyrB, hsp60, recA, rpoB and tuf sequences (4114 bp) and the phylogenomic tree based on whole genome sequences indicated that strains F306-1T and F447 were most closely related to L. kullabergensis Biut2NT, and strain F551-2T was most closely related to L. kimbladii Hma2NT. Strains F306-1T and F447 shared 99.9 % average nucleotide identity (ANI), 99.7 % digital DNA–DNA hybridization (dDDH) and 99.9 % average amino acid identity (AAI) values, indicating that they belong to the same species. Strain F306-1T exhibited the highest ANI (94.4 %), dDDH (56.7 %) and AAI (94.7 %) values to L. kullabergensis Biut2NT. Strain F551-2T had the highest ANI (94.0 %), dDDH (54.3 %) and AAI (95.8 %) values with L. kimbladii Hma2NT. Acid production from amygdalin, maltose, starch, gentiobiose and turanose, activity of esterase (C4) and α-glucosidase, growth with 3 % NaCl at 37 °C under strict anaerobic condition (on mMRS agar plates), and growth with 1–6% NaCl at 37 °C under aerobic condition (on mMRS agar plates supplemented with 0.05 % cysteine or with 1 % cysteine and 2 % fructose) could differentiate strains F306-1T and F447 from L. kullabergensis DSM 26262T. Acid production from d-glucose, arbutin and gentiobiose, growth with 3 % NaCl at 37 °C under strict anaerobic condition (on mMRS agar plates), and growth at 45 °C under strict anaerobic condition (on mMRS agar plates) could differentiate strain F551-2T from L. kimbladii DSM 26263T. Based upon the data obtained in the present study, two novel species, Lactobacillus huangpiensis sp. nov. and Lactobacillus laiwuensis sp. nov., are proposed and the type strains are F306-1T (=LMG 32144T=JCM 34361T=CCTCC AB 2020300T) and F551-2T (=JCM 34502T=CCTCC AB 2021027T), respectively.

An obligately anaerobic, Gram-stain-positive, spore-forming, short-rod-shaped bacterium, designated strain YH-C36aT, was isolated from a pig farm faeces dump. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belongs to the genus Faecalicatena and is most closely related to Faecalicatena contorta KCTC 5831T, Faecalicatena fissicatena KCTC 15010T and Faecalicatena orotica KCTC 15331T, with 96.3, 96.2, and 96.0 % sequence similarity, respectively. The average nucleotide identity values for strain YH-C36aT and the closest related strains were lower than 72 %. The G+C content of the isolate was 43.0 mol%. The cell-wall peptidoglycan was A1γ type and contained meso-diaminopimelic acid. The predominant fatty acids were C16 : 0, C18 : 1 cis 9, C16 : 0 DMA, C18 : 0 DMA and C18:0. The major end products of glucose fermentation were lactate, formate and acetate. Based on its phenotypic, phylogenetic and chemotaxonomic properties, a novel species, named Faecalicatena absiana sp. nov., is proposed for strain YH-C36aT (=KCTC 25106T=NBRC 114768T).

Two facultative anaerobic and facultative alkaliphilic indigo-reducing strains, designated F-1T and F-2, were isolated from indigo fermentation liquor produced from couched woad fermentation-based Indian indigo fermentation fluid. The 16S rRNA gene phylogeny showed that Fundicoccus ignavus WS4937T (99.5%) was the closest neighbour of F-1T. The isolated bacterial cells were Gram-stain-positive and facultative anaerobic coccoids. Strain F-1T grew at between 5 and 37 °C with optimum growth between 28‒32 °C. The isolate grew in a pH range of 7.0‒10.5, with optimum growth between pH 9.0‒10.5. The DNA G+C content was 37.6 mol% (HPLC). The whole-cell fatty acid profile mainly consisted (>10 %) of C16 : 0, C16 : 1 ω9c, C18 : 0 and C18 : 1 ω9c. The digital DNA–DNA hybridization value between strain F-1T and F. ignavus WS4937T was 52.9 %. Based on their physiological and biochemical characteristics, and phylogenetic and genomic data, the isolates can be discriminated from F. ignavus WS4937T. The name Fundicoccus fermenti sp. nov. is proposed. The type strain of this species is F-1T (JCM 34140T=NCIMB 15255T).

A Gram-stain-positive, aerobic, motile, rod-shaped bacterium, designated strain LAM9210T, was isolated from a saline soil sample collected from Lingxian County, Shandong Province, PR China. Analysis of the 16S rRNA gene sequence of the isolate revealed highest sequence similarities to the type strain of Sporosarcina pasteurii NCIMB 8841T (97.6 % sequence similarity). The genomic G+C content was 40.4 mol%. The average nucleotide identity and in silico DNA–DNA hybridization values between strain LAM9210T and the type strain of the most closely related species S. pasteurii NCIMB 8841T were 73.6 and 20.6 %, respectively. Strain LAM9210T was found to grow at 10–40 °C (optimum, 30 °C), at pH 6.0–10.0 (optimum, pH 9.0) and with 0–6 % (w/v) NaCl (optimum, 0.5 %), respectively. The major fatty acids were anteiso-C15 : 0 and iso-C14 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and one unidentified phospholipid. Menaquinone-7 was detected as the predorminant respiratory quinone. Strain LAM9210T contained glycine, lysine, alanine and glutamic acid as the diagnostic amino acids in the cell-wall peptidoglycan. On the basis of phenotypic, phylogenetic and genotypic data, strain LAM9210T is considered to represent a novel species of the genus Sporosarcina , for which the name Sporosarcina jiandibaonis sp. nov. is proposed. The type strain is LAM9210T (=CGMCC 1.18607T=GDMCC 1.2002T=JCM 32514T).

Gram-positive coccoid bacteria were isolated from the nasal cavities of pigs and calves as well as from axillar and inguinal skin regions of pigs. Phylogenetic analysis of seven strains based on complete genome, 16S rRNA, hsp60, dnaJ, rpoB and sodA gene sequences and MALDI-TOF MS profiles revealed that they belonged to the genus Macrococcus with the closest relatedness to Macrococcus canis , Macrococcus caseolyticus subsp. caseolyticus and Macrococcus caseolyticus subsp. hominis . DNA relatedness of the type strain JEK37T with the type strains of M. canis , M. caseolyticus subsp. caseolyticus and M. caseolyticus subsp. hominis was 23.4, 23.1 and 23.0 % by digital DNA–DNA hybridization and 80.39, 80.45 and 80.87 % by average nucleotide identity (ANI) calculations, confirming that they do not belong to the same species. The DNA G+C content of JEK37T was 35.65 mol%. The novel strains can be differentiated from M. canis KM 45013T by the ability to fermentate d-ribose and by the absence of DNAase production and haemolysis, from M. caseolyticus subsp. caseolyticus CCUG 15606T by the ability to fermentate sucrose and from both species by the inability to grow in 9 and 12% NaCl. They differ from M. caseolyiticus subsp. hominis by the presence of α-glucosidase. The most common fatty acids of JEK37T were C14 : 0, C18 : 1 ω9c and C18 : 0. Known polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, aminolipid, aminoglycolipid, aminophospholipid, glycolipid and phospholipid. Cell-wall peptidoglycan of JEK37T was of type A3α l-Lys–Gly2–L-Ser–Gly (similar to A11.3) and the respiratory quinolone was menaquinone 6. Based on their genotypic and chemotaxonomic characteristics, these strains represent a novel species of the genus Macrococcus , for which we propose the name Macrococcus armenti sp. nov. The type strain is JEK37T (=DSM 112712T=CCOS 1982T).

A Gram-stain-variable, motile, aerobic, spore-forming, rod-shaped bacterium, designated as strain LOB 377T, was isolated from soil sampled in Koka County (now Konan City), in Shiga Prefecture, Japan. To determine its taxonomic position, the bacterium was evaluated by a polyphasic approach based on genomic, phenotypic and chemotaxonomic tests. From phylogenetic analysis based on 16S rRNA gene sequences, strain LOB 377T (LC570960) was revealed to have the highest similarity to the type strain of Neobacillus mesonae FJAT-13985T (9.1 %). The average nucleotide identity and digital DNA–DNA hybridization values between both strains based on whole genome sequences were 84.1 and 42.8 %, respectively, which were below the recommended thresholds. The strain grew at 15–45 °C (optimum, 25–37 °C), at pH 5.5–9.5 (optimum, pH 6.5–8.5) and with 1.0–2.0 % (w/v) NaCl. Cell-wall peptidoglycan of the strain contained meso-diaminopimelic acid. The major menaquinone was menaquinone-7. Predominant fatty acids were iso-C15 : 0, anteiso-C15 : 0, iso-C14 : 0 and C16 : 1 ω11c. The polar lipids were phosphatidylglycerol, unidentified phospholipid and unidentified aminophospholipid. The DNA G+C content was 40.5 mol%. According to the genomic, phenotypic and chemotaxonomic results, strain LOB 377T represents a novel species in the genus Neobacillus , for which the name Neobacillus kokaensis sp. nov. is proposed. The type strain is LOB 377T (=ATCC 31382T=NBRC 114637T=DSM 113418T).

Some species of the genus Clostridium are efficient acetate producers and have been deemed useful for upgrading industrial biogas. An acetogenic, strictly anaerobic, Gram-stain-positive, subterminal endospore-forming bacterium designated strain PL3T was isolated from peatland soil enrichments with H2 and CO2. Cells of strain PL3T were 0.8–1.0×4.0–10.0 µm in size and rod-shaped. Growth of strain PL3T occurred at pH 6.0–7.5 (optimum, pH 7.0), at 20–40 °C (optimum, 30 °C) and with 0–1.5 % (w/v) NaCl (optimum, 0.5%). Biochemical analyses revealed that strain PL3T metabolized lactose, maltose, raffinose, rhamnose, lactic acid, sorbitol, arabinose and glycerol. Acetic acid was the predominant metabolite under anaerobic respiration with H2/CO2. The major cellular fatty acids were C16 : 0, C16 : 1 cis 9 and C17 : 0 cyc. The main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, aminolipid and aminophospholipid. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain PL3T belongs to the genus Clostridium with the highest sequence similarity to Clostridium aciditolerans DSM 17425T (98.6 %) followed by Clostridium nitrophenolicum (97.8 %). The genomic DNA G+C content of strain PL3T was 31.1 mol%.The genomic in silico DNA–DNA hybridization value between strain PL3T and C. aciditolerans DSM 17425T was 25.1 %, with an average nucleotide identity of 80.2 %. Based on phenotypic, chemotaxonomic and phylogenetic differences, strain PL3T was suggested to represent a novel species of the genus Clostridium , for which the name Clostridium thailandense sp. nov. is proposed. The type strain is PL3T (=DSM 111812T=TISTR 2984T).