National Institute of Technology and Evaluation

A novel Gram-staining-negative, rod-shaped, aerobic bacterial strain, designated as TSM-5 T, was isolated from surface seawater of a marine inlet. This strain grew well at 10–30 °C, pH 5.0–10.0 and in the presence of 0–4% (w/v) NaCl. Strain TSM-5 T was non-flagellated and positive for catalase and oxidase activities, and the major respiratory quinone was menaquinone-6 (MK-6). The major fatty acids present in the strain were iso-C15:1 G, iso-C15:0, iso-C15:0 3-OH, iso-C17:0 3-OH, and summed feature 3 (comprising C16:1ω7c and/or C16:1ω6c). The major polar lipid of the strain was phosphatidylethanolamine. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain TSM-5 T shared 92.6 to 94.0% sequence similarity with valid species of the genus Cellulophaga, and the phylogenetic branch of this strain divided the genus into two groups. Based on the average nucleotide identity (ANI), average amino acid identity (AAI), and digital DNA-DNA hybridization (dDDH) with genomes from the closely related family Flavobacteriacea, strain TSM-5 T represents a lineage distinct from genus Cellulophaga, suggesting that this genus should be divided into two genera. Physiological and biochemical comparisons between strain TSM-5 T and closely related species revealed additional differences, including optimal NaCl concentrations and pH for growth, as well as metabolic traits such as flexirubin pigment reaction. Based on these results, we propose a novel species in a new genus, Allocellulophaga tsushimaensis gen. nov. sp. nov., with TSM-5 T (= KCTC 52508 T = NBRC 112430 T) as the type strain. Furthermore, we propose that Cellulophaga baltica, Cellulophaga pacifica, Cellulophaga tyrosinoxydans, and Cellulophaga algicola, currently valid species of the genus Cellulophaga, be reclassified into a novel genus, Paracellulophaga gen. nov., with Paracellulophaga baltica NN015840T (= DSM 24729 T = ATCC 700862 T) as the type species.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10482-026-02275-0.

A crude oil aggregation-forming, strictly anaerobic, Gram-stain-positive, spore-forming, rod-shaped, motile and mesophilic bacterium, named strain SH18-2T, was isolated from marine sediment near Sado Island in the Sea of Japan. The temperature, salinity and pH ranges of this strain for the growth were 15-40 °C (optimum 35 °C), 0.5-6.0% NaCl (w/v; optimum 2.0%) and pH 6.0-8.0 (optimum 7.5), respectively. A phylogenetic analysis of the 16S rRNA gene sequence showed that this strain belongs to the genus Clostridium. It was most closely related to Clostridium grantii A-1T (97.02% homology). The draft genome size was 5616089 bp, with a G+C content of 29.7 mol%. The digital DNA-DNA hybridization values determined using the Genome-to-Genome Distance Calculator and the average nucleotide identity between this strain and C. grantii A-1T were 20.30 and 78.24%, respectively. Whole-organism hydrolysates of strain SH18-2T contained meso-diaminopimelic acid; the major fatty acid was C16:0, and the polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, glycolipid and three unidentified polar lipids. From the results of genotypic, phenotypic and chemotaxonomic analyses, SH18-2T (=NBRC 116030T=DSM 115762T) is the type strain and represents a novel species of the genus Clostridium, and the name Clostridium sediminicola sp. nov. is proposed.

Two Gram-stain-negative, straight rods, non-motile, asporogenous, catalase-negative and obligately anaerobic butyrate-producing strains, HLW78T and CYL33, were isolated from faecal samples of two healthy Taiwanese adults. Phylogenetic analyses of 16S rRNA and DNA mismatch repair protein MutL (mutL) gene sequences revealed that these two novel strains belonged to the genus Faecalibacterium. On the basis of 16S rRNA and mutL gene sequence similarities, the type strains Faecalibacterium butyricigenerans AF52-21T(98.3–98.1 % and 79.0–79.5 % similarity), Faecalibacterium duncaniae A2-165T(97.8–97.9 % and 70.9–80.1 %), Faecalibacterium hattorii APC922/41-1T(97.1–97.3 % and 80.3–80.5 %), Faecalibacterium longum CM04-06T(97.8–98.0% and 78.3 %) and Faecalibacterium prausnitzii ATCC 27768T(97.3–97.4 % and 82.7–82.9 %) were the closest neighbours to the novel strains HLW78T and CYL33. Strains HLW78T and CYL33 had 99.4 % both the 16S rRNA and mutL gene sequence similarities, 97.9 % average nucleotide identity (ANI), 96.3 % average amino acid identity (AAI), and 80.5 % digital DNA–DNA hybridization (dDDH) values, indicating that these two strains are members of the same species. Phylogenomic tree analysis indicated that strains HLW78T and CYL33 formed an independent robust cluster together with F. prausnitzii ATCC 27768T. The ANI, AAI and dDDH values between strain HLW78T and its closest neighbours were below the species delineation thresholds of 77.6–85.1 %, 71.4–85.2 % and 28.3–30.9 %, respectively. The two novel strains could be differentiated from the type strains of their closest Faecalibacterium species based on their cellular fatty acid compositions, which contained C18 : 1 ω7c and lacked C15 : 0 and C17 : 1 ω6c, respectively. Phenotypic, chemotaxonomic and genotypic test results demonstrated that the two novel strains HLW78T and CYL33 represented a single, novel species within the genus Faecalibacterium, for which the name Faecalibacterium taiwanense sp. nov. is proposed. The type strain is HLW78T (=BCRC 81397T=NBRC 116372T).

LC/MS/MSによる陰イオン界面活性剤の定量分析の検討を行った．検討の結果，分析対象の陰イオン界面活性剤が，バックグラウンドとして，移動相中に高濃度に検出し，定量分析を妨害することを確認した．そこで，六方切換バルブに前処理用カラムを取り付け，定量分析の妨害となる陰イオン界面活性剤のバックグラウンドを低減することを可能としたLC/MS/MSを構築した．構築したLC/MS/MSによる陰イオン界面活性剤の定量方法を検討し，検量線作成の結果，良好な直線性を示し，定量が可能であることが確認された．

Three bacterial strains, designated SSBR10-3T, SSTM10-2T and SSHM10-5T, were isolated from saltern soil sampled in Jeollanam-do, Republic of Korea. Cells were aerobic, Gram-stain-positive, flagellated and rod-shaped. The strains grew optimally at 28°C and at pH 7.0. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains SSBR10-3T, SSTM10-2T and SSHM10-5T were placed within the genus Halobacillus, showing the highest similarity to Halobacillus alkaliphilus FP5T (98.6 %), 'Halobacillus ihumii' Marseille-Q1234T (98.5 %) and Halobacillus locisalis MSS-155T (98.6 %), respectively. The genomic similarity values between strains SSBR10-3T, SSTM10-2T and SSHM10-5T and their related species were 17.6-22.6 % for digital DNA-DNA hybridization (dDDH) and 69.6-78.5 % for orthologous average nucleotide identity (OrthoANI), which were lower than the thresholds recommended for species delineation. The dDDH and OrthoANI values among the three strains were below 38.3 and 89.4 %, respectively. Besides the differences in genomic features, strains SSBR10-3T, SSTM10-2T and SSHM10-5T were distinct from each other and from members of the genus in terms of phenotypic traits related to substrate assimilation. The cell-wall peptidoglycan contained meso-diaminopimelic acid, the major fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0, and the predominant menaquinone was MK-7 for all three strains. Diphosphatidylglycerol, phosphatidylglycerol and an unidentified phospholipid were present in their polar lipid profiles. Based on a polyphasic approach incorporating genomic data, strains SSBR10-3T, SSTM10-2T and SSHM10-5T represent novel species, for which the names Halobacillus salinarum sp. nov. (SSBR10-3T=DSM 114353T=KACC 21935T=NBRC 115504T), Halobacillus shinanisalinarum sp. nov. (SSTM10-2T=DSM 114354T=KACC 21936T=NBRC 115505T) and Halobacillus amylolyticus sp. nov. (SSHM10-5T=DSM 114355T= KACC 21937T=NBRC 115506T) are proposed.

A novel bacterium, designated 5-21aT, isolated from chitin-treated upland soil, exhibits methionine (Met) auxotrophy and chitinolytic activity. A physiological experiment revealed the cobalamin (synonym, vitamin B12)(Cbl)-auxotrophic property of strain 5-21aT. The newly determined complete genomic sequence indicated that strain 5-21aT possesses only the putative gene for Cbl-dependent Met synthase (MetH) and lacks that for the Cbl-independent one (MetE), which implies the requirement of Cbl for Met-synthesis in strain 5-21aT. The set of genes for the upstream (corrin ring synthesis) pathway of Cbl synthesis is absent in the genome of strain 5-21aT, which explains the Cbl-auxotrophy of 5-21aT. This strain was characterized via a polyphasic approach to determine its taxonomic position. The nucleotide sequences of two copies of the 16S rRNA gene of strain 5-21aT indicated the highest similarities to Lysobacter soli DCY21T(99.8 and 99.9 %) and Lysobacter panacisoli CJ29T(98.7 and 98.8 %, respectively), whose Cbl-auxotrophic properties were revealed in this study. The principal respiratory quinone was Q-8. The predominant cellular fatty acids were iso-C15:0, iso-C16:0 and iso-C17:1 ω9c. The complete genome sequence of strain 5-21aT revealed that the genome size was 4 155 451 bp long and the G+C content was 67.87 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between strain 5-21aT and its most closely phylogenetic relative L. soli DCY21T were 88.8 and 36.5%, respectively. Based on genomic, chemotaxonomic, phenotypic and phylogenetic data, strain 5-21aT represents a novel species in the genus Lysobacter, for which the name Lyobacter auxotrophicus sp. nov. is proposed. The type strain is 5-21aT (=NBRC 115507T=LMG 32660T).

Three bacterial strains, H21R-40T and H21R-36 from garlic (Allium sativum) and H25R-14T from onion (Allium cepa), were isolated from plant rhizospheres sampled in the Republic of Korea. Results of 16S rRNA gene sequence analysis revealed the highest sequence similarity of strain H21R-40T to Leucobacter celer subsp. astrifaciens CBX151T (97.3 %) and Leucobacter triazinivorans JW-1T (97.2 %), and strain H25R-14T to Leucobacter insecticola HDW9BT (98.8 %) and Leucobacter humi Re6T (98.4 %), while the sequence similarity between strains H21R-40T and H21R-36 was 99.8 %. According to the phylogenomic tree, strains H21R-40T with H21R-36 formed an independent clade separable from other Leucobacter species within the genus Leucobacter and strain H25R-14T clustered with Leucobacter insecticola HDW9BT, Leucobacter coleopterorum HDW9AT and Leucobacter viscericola HDW9CT. Strains H21R-40T and H21R-36 had orthologous average nucleotide identity (OrthoANI) and digital DNA-DNA hybridization (dDDH) values (98.1 % and 86.9 %, respectively) higher than the threshold ranges for species delineation (95-96 % and 70 %, respectively). The OrthoANI and dDDH values between two strains (H21R-40T and H25R-14T) and the type strains of species of the genus Leucobacter were lower than 81 and 24 %, respectively. The peptidoglycan type of three strains was type B1. The major menaquinones and major polar lipids of the strains were MK-11 and MK-10, and diphosphatidylglycerol, phatidylglycerol and an unidentified glycolipid, respectively. The major fatty acids (more than 10 % of the total fatty acids) of strains H21R-40T and H21R-36 were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0, and those of strain H25R-14T were anteiso-C15 : 0 and iso-C16 : 0. The phenotypic, chemotaxonomic and genotypic data obtained in this study showed that the strains represent two novel species of the genus Leucobacter, named Leucobacter allii sp. nov. (H21R-40T and H21R-36) and Leucobacter rhizosphaerae sp. nov. (H25R-14T). The respective type strains are H21R-40T (=DSM 114348T=JCM 35241T=KACC 21839T=NBRC 115481T) and H25R-14T (=DSM 114346T=JCM 35239T=KACC 21837T=NBRC 115479T).

ABSTRACTThe marine Streptomyces sp. strain GMY01 was isolated from Indonesian marine sediment. Genome mining analysis revealed that GMY01 has 28 biosynthetic gene clusters, dominated by genes encoding nonribosomal peptide synthetase and polyketide synthase.