DNA-DNA Hybridization Values Research Articles

Cerasicoccus fimbriatus sp. nov., isolated from the mid-ridge of the Southwest Indian Ocean.

A Gram-stain-negative bacterium, designated as strain TK19100T, was isolated from the mid-ridge of the Southwest Indian Ocean. Cells of strain TK19100T were strictly aerobic, non-motile and short-rod shaped with fimbriae-like structures around the cell surface. Growth occurred at 15-40°C, at pH 6.0-9.0 and with 1-10% (w/v) NaCl. Strain TK19100T shared the highest 16S rRNA gene sequence similarity with Cerasicoccus frondis NBRC 105381T of 97.74%, followed by Cerasicoccus arenae KCTC 12870T of 97.69% and Cerasicoccus maritimus NBRC 105382T of 97.40%. The phylogenetic tree based on both 16S rRNA genes and 92 core genes placed strain TK19100T in a new linage within the genus Cerasicoccus. The genome size of strain TK19100T was 5.07Mb with the DNA G + C content 56.1%. The average nucleotide identity value and the digital DNA-DNA hybridization value of strain TK19100T compared with the closest related species, Cerasicoccus frondis, were 78.39% and 23.70%, respectively. Strain TK19100T encoded a previously unreported combination of GH16 and Carbohydrate Binding Module 96. The major fatty acids of strain TK19100T were C14:0 and C18:1 ω9c. Menaquinone-7 was the sole respiratory quinone. The phenotypic and genotypic characterization analysis indicate that strain TK19100T represents a novel species affiliated to the genus Cerasicoccus, for which the name Cerasicoccus fimbriatus sp. nov. is proposed. The type strain is TK19100T (= CGMCC 1.18957T = NBRC 116189T).

A description of Joostella sp. strain CR20 with potential biotechnological applications.

The underexplored halophilic genus Joostella within the Flavobacteriaceae family consists of only two species, both of which have received little attention for their potential biotechnological applications. In this study, we report the isolation and characterisation of a novel halophilic bacterium, strain CR20, using a genomic approach to investigate its biotechnological potential. Analysis of the 16S rRNA gene revealed that strain CR20 shares 97.5% and 96.2% sequence similarity with Joostella marina DSM 19592T and Joostella atrarenae M1-2T, respectively. Strain CR20 exhibited average nucleotide identity and digital DNA-DNA hybridisation values of 76.8-79.1% and 20.8-22.8%, respectively, with Joostella spp., which fall below the species delineation thresholds. Additionally, strain CR20 demonstrated average amino acid identity and percentage of conserved proteins values of 81.3-84.0% and 71.7-75.3%, respectively, with Joostella spp., above the genus delineation thresholds. Meanwhile, the average amino acid identity and percentage of conserved proteins values of strain CR20 against Galbibacter spp. are 73.9-80.0% and 61.3-72.3%, respectively, also above the genus delineation thresholds. These findings indicated strain CR20 has a close relationship with both genera. Chemotaxonomic analysis of strain CR20 identified predominant fatty acids, including iso-C17:0 3OH (25.3%), iso-C15:0 (14%), and C16:1 ω6c/C16:1 ω7c (12.2%). The assembled genome comprises 62 contigs, with a size of approximately 3,168,727bp and a G + C content of 35.1%. Among 2,804 predicted genes, 2,559 were classified into 25 COG functional groups. A total of 68 genes with potential industrial applications were identified, including 1 β-mannanase, 2 β-xylosidases, 1 polysaccharide deacetylase, 4 other hemicellulases, 6 β-glucosidases, 25 proteases, and 29 phosphate-solubilising enzymes. Hydrolytic assays confirmed that strain CR20 produces these enzymes extracellularly. These findings highlight strain CR20 has potential for industrial applications.

Jannaschia ovalis sp. nov., a marine bacterium isolated from a tidal flat sediment in the Republic of Korea.

A Gram-stain-negative, non-motile, aerobic, pale-yellow coloured and oval-shaped bacterium designated GRR-S6-38T was isolated from a tidal flat sediment that collected from Garorim Bay of the Yellow Sea, Republic of Korea. Strain GRR-S6-38T grew at 15-40°C (optimum, 30°C) at pH 6-9 (optimum, pH 7) and at 2-6% (w/v) NaCl (optimum, 2% NaCl). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain GRR-S6-38T was closely related to those of J. seosinensis CL-SP26T (98.1%), J. helgolandensis DSM 14858T (97.4%), J. rubra CECT 5088T (97.4%), J. faecimaris DSM 100420T (97.4%), J. marina SHC163T (97.3%), J. donghaensis CECT 7802T (97.2%) and J. pohangensis DSM 19073T (97.1%). The average nucleotide identity and digital DNA-DNA hybridization value between GRR-S6-38T and related type strains were 71.47-78.59% and 17.80-21.40%. Strain GRR-S6-38T was characterized as having Q-10 as the predominant respiratory quinone and the major principle fatty acids (> 10%) were SF 8 (C18:1 ω7c/C18:1 ω6c, 39.3%), C19:0 ω8c cyclo (13.2%) and C18:1 ω7c 11-methyl (10.6%). The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, an unidentified lipid, two unidentified aminolipids and two unidentified phospholipids. The assembled genome of strain GRR-S6-38T has a total length of 3.32 Mbp, and a G + C content is 69.98%. Based on the polyphasic taxonomic evidence presented in this study, GRR-S6-38T is regarded to represent a novel species within the genus Jannaschia, for which name Jannaschia ovalis sp. nov. is proposed. The type strain is GRR-S6-38T (= KCTC 82518 = KACC 22240 = JCM 36187).

Pseudomonas machongensis sp. nov. and Stenotrophomonas capsici sp. nov., isolated from wilted pepper plants.

Two Gram-stain-negative, aerobic, rod-shaped, non-endospore-forming bacteria, designated as strain MH1T and MH2T, were isolated from branches of wilted pepper plants (Capsicum annuum) collected from a farmland in Machong town, Guangdong, China, and investigated using a polyphasic approach. MH1T grew at temperatures of 4-42 °C (optimum 28 °C), with 0-6.0 % (w/v) NaCl and at pH 4.0-10.0 (optimum pH 4.0). MH2T grew at temperatures of 4-42 °C (optimum 28 °C), with 0-6.0% (w/v) NaCl and at pH 4.0-10.0 (optimum pH 5.0). Analysis of the 16S rRNA gene sequence indicated that MH1T belongs to Stenotrophomonas and MH2T belongs to Pseudomonas. Genome-based phylogenetic analysis further established that MH1T shares the closest evolutionary relationships with Stenotrophomonas humi DSM 18929 and Stenotrophomonas terrae DSM 18941, and MH2T is sister to Pseudomonas wayambapalatensis RW3S1. Whole-genome comparisons between MH1T and known Stenotrophomonas species revealed average nucleotide identity (ANI) values up to 84.5%, as well as digital DNA-DNA hybridization (dDDH) values up to 28.3%, both substantially lower than the accepted thresholds for species delineation (ANI: 95%; dDDH: 70%). The ANI and dDDH values between MH2T and known Pseudomonas species were at most 94.6 and 59.2 %, respectively. Additional biochemical and physiological analyses further support that MH1T and MH2T represent a novel species in Stenotrophomonas and Pseudomonas, respectively. Notably, the differences in carbon source utilization could differentiate MH1T and its close relatives in Stenotrophomonas. The major fatty acids were iso-C15 : 0, iso-C16 : 0 and iso-C14 : 0 for MH1T and were C16 : 0, C18 : 1 ω7c/C18 : 1 ω6c (summed feature 8), C16 : 1 ω7c/C16 : 1 ω6c (summed feature 3) and C17 : 0 cyclo for MH2T. Therefore, we propose a new species Stenotrophomonas capsici sp. nov., with MH1T (=GDMCC 1.3749T=JCM 36317T) as the type strain, and a new species Pseudomonas machongensis sp. nov., with MH2T (=GDMCC 1.3750T=JCM 36318T) as the type strain. The MH1T genome has a size of 4.18 Mb and a GC-content of 67.19 mol%, while the MH2T genome has a size of 5.71 Mb and a GC-content of 63.12 mol%.

Altererythrobacter arenosus sp. nov., Isolated from Marine Sediment.

Strain CAU 1644T was isolated from sea sand at Ganghwa Island, Republic of Korea. The bacterium is aerobic, Gram-staining negative, yellow-colored, non-motile, catalase-positive, and rod-shaped. Cells of strain CAU 1644T grew at 20-40°C and pH 6.0-10.0 with 0%-3.0% (w/v) NaCl. The 16S rRNA gene sequences of Altererythrobacter epoxidivorans CGMCC 1.7731T (identity, 98.5%) and A. xiamenensis CGMCC 1.12494T (98.0%) had the highest similarities to strain CAU 1644T. Strain CAU 1644T exhibits affiliation with the genus Altererythrobacter by phylogenetic trees based on the 16S rRNA gene and core-genome sequences. The predominant quinone was ubiquinone-10 (Q-10) and the major fatty acids were summed feature 8 (C18:1ω7c and/or C18:1ω6c) and summed feature 3 (C16:1ω7c and/or C16:1ω6c). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values of strain CAU 1644T were 77.6% and 20.1% with A. epoxidivorans CGMCC 1.7731T, respectively, and 77.4% and 21.0% with A. xiamenensis CGMCC 1.12494T. The G + C content is 62.5%. Based on phenotypic and genotypic analyses, strain CAU 1644T represents a novel species of the genus Altererythrobacter, for which the name Altererythrobacter arenosus sp. nov., is proposed. The type strain is CAU 1644T (= KCTC 92082T = MCCC 1K07086T).

Genomic profiling and assessment of the plant growth-enhancing traits in the novel actinomycete, Plantactinospora soli sp. nov.

Strain KLBMP 9567T, an isolate from weathered soil, was identified as a new actinobacterial species through a comprehensive polyphasic approach. Phylogenetic evaluations relying on 16S rRNA gene sequence placed KLBMP 9567T within the genus Plantactinospora, alongside its close relatives Plantactinospora veratri NEAU-FHS4T and Plantactinospora sonchi NEAU-QY2T, both sharing 98.6% sequence similarity. However, KLBMP 9567T demonstrated low digital DNA-DNA hybridization values with P. veratri NEAU-FHS4T and P. sonchi NEAU-QY2T, recorded at 48.5 and 29.1%, respectively. Meanwhile, the average nucleotide identity values were correspondingly recorded at 91.1 and 83.1%. The cell wall of KLBMP 9567T contained meso-diaminopimelic acid, with xylose, mannose, glucose and galactose as diagnostic sugars. The diagnostic dominant phospholipids included diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. Given phenotypic and genotypic data, KLBMP 9567T is recognized as a new species within the Plantactinospora genus, named Plantactinospora soli sp. nov., with the type strain KLBMP 9567T (= CGMCC 4.7773T = NBRC 115787T). Genome mining revealed genes associated with plant growth promotion, specifically those encoding enzymes for synthesizing plant hormones like indole acetic acid and gibberellic acid. Experimental validation demonstrated that KLBMP 9567T can produce significant quantities of indole acetic acid (20.6 mg l-1) and gibberellic acid (42.6 mg l-1). Moreover, co-culture with Arabidopsis showed marked growth enhancements: a 56.3% increase in lateral root proliferation, a 16.7% elongation in primary root length and a 44.8% boost in biomass accumulation. These findings underscore the strain's potent plant growth-promoting attributes, providing profound insights into the mechanisms by which KLBMP 9567T enhances plant growth.

Xenorhabdus bharatensis sp. nov., Xenorhabdus entomophaga sp. nov., Xenorhabdus siamensis sp. nov., and Xenorhabdus thailandensis sp. nov. Isolated from Steinernema Entomopathogenic Nematodes

Four Gram-stain-negative bacterial strains, CS20T, AUT15.5T, XENO-11T, and CCN3.3T, isolated from Steinernema entomopathogenic nematodes, were found to represent novel species within the genus Xenorhabdus (Gammaproteobacteria, Morganellaceae). In this study, we described these new species using whole-genome phylogenomic reconstructions, sequence identity values from core genome sequences, and phenotypic characterization. Phylogenetic reconstructions based on 16S rRNA gene sequences showed that: (i) strain CS20T is closely related to X. stockiae DSM 17904T, (ii) strain AUT15.5T is closely related to X. budapestensis DSM 16342T, (iii) strain XENO-11T is closely related to X. khoisanae DSM 25463T, and (iv) strain CCN3.3T is closely related to X. griffiniae DSM 17911T. The 16S rRNA gene sequence similarity value between strain CS20T and X. stockiae DSM 17904T is 97.8%, between strain AUT15.5T and X. budapestensis DSM 16342T is 98.1%, between strain XENO-11T and X. khoisanae DSM 25463T is 97.8%, and between strain CCN3.3T and X. griffiniae DSM 17911T is 98.6%. Phylogenomic reconstructions using whole-genome sequences showed that: (i) strain CS20T is closely related to X. stockiae DSM 17904T and X. innexi DSM 16336T, (ii) strain AUT15.5T is closely related to X. indica DSM 17382T, (iii) strain XENO-11T is closely related to X. khoisanae DSM 25463T, and (iv) strain CCN3.3T is closely related to X. griffiniae DSM 17911T. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strains CS20T, AUT15.5T, XENO-11T, and CCN3.3T and the type strains of their more closely related species are below the 70% and the 95–96% divergence thresholds, respectively, used for prokaryotic species delineation. Hence, we propose the following four new species: Xenorhabdus bharatensis sp. nov. (the type strain is CS20T=CCM 9320T=CCOS 2070T), X. entomophaga sp. nov. (the type strain is XENO-11T=CCM 9389T=CCOS 2111T), X. siamensis sp. nov. (the type strain is AUT15.5T=CCM 9405T=CCOS 2116T), and X. thailandensis sp. nov. (the type strain is CCN3.3T=CCM 9406T=CCOS 2115T). The following biochemical tests may be useful for differentiating the novel species from their more closely related taxa: acetoin production, arginine dihydrolase, citrate utilization, gelatinase, glucose oxidation, indole production, and tryptophan deaminase. Our study contributes to a better understanding of the biodiversity and phylogenetic relationships of entomopathogenic bacteria associated with insect parasitic nematodes.

Nocardioides xinjiangensis sp. nov., a novel species isolated from desert soil.

Two novel strains, SYSU D00514T and SYSU D00778, were isolated from desert soil in the Gurbantunggut Desert, Xinjiang Uygur Autonomous Regions, PR China. SYSU D00514T and SYSU D00778 were aerobic, Gram-stain-positive, rod-shaped, catalase-positive and oxidase-negative. SYSU D00514T grew at temperatures ranging from 4 to 37°C (optimum, 28-30°C), at pH 6.0-8.0 (optimum, 7.0), and tolerated NaCl concentrations from 0 to 7.5% (optimum, 0.5% w/v). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains SYSU D00514T and SYSU D00778 were closely related to the members of the genus Nocardioides. Two isolates were most closely related to Nocardioides alpinus Cr7-14T (98.05% and 98.19%). The G + C content of the genomic DNA of strains SYSU D00514T and SYSU D00778 were both 72.8%. Strain SYSU D00514T and the type strains of the genus Nocardioides in the analysis had average nucleotide identity values of 76.8-85.8% as well as digital DNA-DNA hybridization values between 19.5% and 28.0%. The predominant cellular fatty acids of strain SYSU D00514T were C17:1 ω8c, C18:1 ω9c and iso-C16:0. For strain SYSU D00514T, the major polar lipids encompassed diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, an unidentified aminophospholipid and two unidentified aminolipids. The main respiratory quinone of strain SYSU D00514T was MK-8 (H4). Based on the results of polyphasic taxonomic experiments, we propose that strains SYSU D00514T and SYSU D00778 represent a novel species of the genus Nocardioides, with the name Nocardioides xinjiangensis. The type strain is SYSU D00514T (= CGMCC 1.18622T = MCCC 1K05001T = KCTC 49488T).

Veillonella orientalis sp. nov., an anaerobic Gram-stain-negaitve coccus isolated from saliva of a Thai child

ObjectiveThe genus Veillonella is one of the major and important constitutions of the oral microbiome.A novel anaerobic, Gram-strain-negative coccus belonging to the genus Veillonella was isolated from the saliva of a Thai child coming from the east of Thailand. In the present study, the characterization of one strain, S12025-13T is described with the comparison of established species of the genus Veillonella and proposed a novel oral species of the genus Veillonella. MethodsStrain S12025-13T was conclusively identified using phenotypic, phylogenetic, genomic and biochemical approach. In phylogenetic analysis, multi-locus species trees (MLST) analysis was also conducted in this study. ResultsThe strain S12025-13T showed typical phenotypic characteristics of members of the genus Veillonella. Under anaerobic conditions, the strain produced acetic acid and propionic acid as metabolic end products in a trypticase-yeast extract-heamin medium containing 1% (w/v) glucose, 1% (w/v) fructose and 1% (v/v) sodium lactate. Sequencing of their 16S rRNA genes confirmed that it belongs to the genus Veillonella. Comparative analysis of the 16S rRNA, dnaK and rpoB gene sequences indicated that phylogenetically the strain comprised a distinct novel branch within the genus Veillonella. The novel strain showed 99.77, 97.29, and 99.02% similarity to partial 16S rRNA, dnaK and rpoB gene sequencing, respectively, to the type strains of the most closely related species. In MLST (selected 79 genes) analysis, strain S12025-13T formed a distinct taxon with robust bootstrap support (100%) within the genus Veillonella. Furthermore, strain S12025-13T shared the highest average nucleotide identity (ANI) value (95.97%) with the type strain of the most closely related species, “Veillonella faecalis” which is not validly published under the International Code of Nomenclature of Prokaryotes (ICNP). Likewise, strain S12025-13T showed the highest digital DNA-DNA hybridization (dDDH) value (65.00%) with the type strain of “V. faecalis”. In addition, strain S12025-13T formed a distinct branch in this clade with a bootstrap value of 72% with Veillonella nakazawae and 51% with “V. faecalis” related species of the genus Veillonella in the genome based phylogenetic tree. ConclusionThe novel strain could be discriminated from previously reported species of the genus Veillonella based on partial dnaK gene sequencing, MLST analysis, ANI and dDDH values, and genome-based phylogeny. Based on these observations, this strain represents a novel species, for which the name Veillonella orientalis sp., nov. is proposed. The type strain is S12025-13T (= JCM 33967T = CCUG 74596T).

Methylocystis borbori sp.nov., a novel methanotrophic bacterium from the sludge of a freshwater lake and its metabolic properties.

A novel methanotrophic strain 9NT was isolated from the sludge of a freshwater lake. Cells were aerobic, Gram-stain-negative, non-motile pleomorphic rods with intracytoplasmic membrane systems that appropriate type-II methanotrophs and hemispherical and spherical exocellular formations on the perimeter of the cell wall surface. The novel isolate grows only on methane or methanol as the sole carbon and energy source, at 10-37°C (optimum 28-30°C), pH 4.5-9.0 (optimum 7.0-7.5), up to 1% NaCl (optimum 0.3-0.5%). Methanol supported the growth of the strain 9NT in a wide range of concentrations from 0.05 to 5.0% (v/v) with an optimum of 0.5% (v/v). The major fatty acids were C18:1ω8c and C18:1ω7c. Based on 16S rRNA gene sequence phylogenetic analysis, strain 9NT was closely related to representatives of the genus Methylocystis (96.5-98.3%). The genome of strain 9NT was 3.34Mbp in size with 63.5% of G + C content. The average nucleotide identity and digital DNA-DNA hybridization values between strain 9NT and closely related type strains of genus Methylocystis were 78.0-82.4% and 20.9-24.3%, respectively. The careful genome annotation of the novel strain shows it possessed genes for the detoxification of arsenate and cyanides as well as genes potentially involved in plant growth promotion (such as biosynthesis of indoles, cytokinins, polyhydroxybutyrate, siderophore production, and nitrogen fixation). Based on the phylogenetic, phenotypic, chemotaxonomic and genomic data, we propose Methylocystis borbori sp. nov. as novel species of the genus Methylocystis. The type strain is 9NT (= VKM B-3616T = KCTC 92566T).

Luteimonas flava sp. nov. and Aquilutibacter rugosus gen. nov., sp. nov., isolated from freshwater environments in China and re-examining the taxonomic status of genera Luteimonas and Lysobacter.

Three Gram-stain-negative, aerobic, short rod-shaped and motile strains (FXH3WT, SHGZ20W and SMYT11WT) were isolated from freshwater environments in China. Comparisons based on the 16S rRNA gene sequences indicated that strains FXH3WT and SHGZ20W showed the highest 16S rRNA gene sequence similarity of about 99.6% to 'Luteimonas cellulosilyticus' MIC 1.5, and strain SMYT11WT showed the highest 16S rRNA gene sequence similarity of 99.8% to Luteimonas fraxinea D4P002T, respectively. Observing the phylogenetic trees reconstructed based on the 16S rRNA gene sequences, the species of genera Luteimonas and Lysobacter were not monophyletic and often mixed together. The further reconstructed phylogenomic tree and Genome Taxonomy Database also showed that the species of both genera were polyphyletic, implying that the current taxonomic status of the species of both genera was questionable. The calculated OrthoANIu, digital DNA-DNA hybridization and average amino acid sequence identity (AAI) values supported that strains FXH3WT and SHGZ20W should belong to the same novel species and strain SMYT11WT should also represent an independent novel species. Combining the AAI values and phylogenomic analyses, the species of genera Luteimonas and Lysobacter should be reassigned to 12 genera (Luteimonas, Lysobacter, Cognatiluteimonas, Noviluteimonas, Pseudoluteimonas, Solilutibacter, Agrilutibacter, Cognatilysobacter, Marilutibacter, Novilysobacter, Montanilutibacter and Aerolutibacter). The AAI values 69.5-76.0% were also proposed as the Lysobacteraceae-specific thresholds for genus delineation. Strain SMYT11WT should represent a novel species of the genus Luteimonas, for which the name Luteimonas flava sp. nov. (type strains SMYT11WT=GDMCC 1.4275T=KCTC 8304T) is proposed. Strains FXH3WT and SHGZ20W should represent a novel species of a new genus Aquilutibacter rugosus gen. nov., sp. nov. The type strain of the type species is FXH3WT (=GDMCC 1.4096T=KCTC 8154T).

Campylobacter sputorum subsp. bovis subsp. nov., isolated from cattle, and an emended description of Campylobacter sputorum.

Six urease-negative Campylobacter strains were isolated from cattle faeces over a 19-month period from 2009 to 2010. These strains were initially identified as Campylobacter sputorum by 16S rRNA gene and atpA typing. Initial studies characterizing these strains by multilocus sequence typing and genome sequencing further supported their classification as C. sputorum but indicated that these strains form a divergent clade within the species. A polyphasic study was undertaken here to clarify their taxonomic position. Phylogenetic analyses were performed based on 16S rRNA gene sequences and the concatenated sequences of 330 core genes, with the latter analysis also placing the six strains into a clade distinct from the three C. sputorum biovars. Pairwise digital DNA-DNA hybridization values identified these strains as C. sputorum, and the pairwise average nucleotide identity values were consistent with those observed between current Campylobacter subspecies pairs. Standard phenotypic testing was also performed. All strains are microaerobic, anaerobic, motile, Gram-negative and oxidase- and catalase-positive; cells are curved rods or spirals. Strains can be distinguished from the C. sputorum biovars by the presence of alkaline phosphatase activity and triphenyltetrazolium chloride reduction and absence of nitrate reduction. The data presented here show that these strains represent a novel subspecies within C. sputorum, for which the name C. sputorum subsp. bovis subsp. nov. (type strain RM8705T=LMG 32300T=CCUG 75470T) is proposed.

Description of Olsenella absiana sp. nov., an anaerobic bacterium isolated from pig faeces.

A short-rod-shaped, obligate anaerobic, Gram-positive bacterium isolated from the faeces of a pig was designated as the strain YH-ols2223T. Analysis of the 16S rRNA gene sequence revealed that the isolate was most closely related to Olsenella urininfantis KCTC 25755T with 97.7% similarity. The multi-locus sequence tree revealed that the isolate formed a sub-cluster within the genus Olsenella. The average nucleotide identity and the digital DNA-DNA hybridization values for the isolate and the most closely related strains were 74.9-76.6% and 20.8-22.5%, respectively. The end products of the fermentation were lactic acid and acetic acid. The main fatty acids identified were C18 : 0 DMA, C18 : 1 ω9c DMA and C18 : 1 ω9c. The cell wall contained the meso-diaminopimelic acid-based peptidoglycan. Given the chemotaxonomic, phenotypic and phylogenetic properties, YH-ols2223T (= KCTC 25800T = NBRC 116680T) represents a novel taxon. The name Olsenella absiana sp. nov. was proposed.

Roseovarius phycicola sp. nov. and Roseovarius rhodophyticola sp. nov., isolated from marine red algae.

Two Gram-stain-negative, strictly aerobic, non-motile, rod-shaped bacteria, designated as strains S88T and W115T, exhibiting catalase- and oxidase-positive reactions, were isolated from marine red algae in South Korea. Strain S88T exhibited growth at 20-30 °C, pH 6.0-9.0 and 2.0-5.0% (w/v) NaCl, while strain W115T grew at 20-30 °C, pH 7.0-9.0 and 2.0-5.0% (w/v) NaCl. Strain S88T contained summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), C16 : 0 and C16 : 0 2-OH as major fatty acids (>5%), with major polar lipids being phosphatidylglycerol, phosphatidylcholine, an unidentified phospholipid, an unidentified aminolipid and two unidentified lipids. Strain W115T contained summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and C16 : 0 as major fatty acids (>5%), with major polar lipids including phosphatidylglycerol, phosphatidylcholine, an unidentified phospholipid, an unidentified aminolipid and unidentified lipids. Ubiquinone-10 was the sole respiratory quinone in both strains, and the genomic DNA G+C contents were 57.0% for strain S88T and 56.5% for strain W115T. Despite 99.86% 16S rRNA gene sequence similarity, strains S88T and W115T shared 88.8% average nucleotide identity (ANI) and 36.8% digital DNA-DNA hybridization (dDDH) value, indicating different species. Phylogenetic and phylogenomic analyses based on 16S rRNA gene and genome sequences, respectively, revealed that strains S88T and W115T formed a phylogenetic lineage within the genus Roseovarius. ANI and dDDH values of both strains with other type strains were less than 73.7 and 20.3%, respectively, confirming that they represent novel species. Based on phenotypic, chemotaxonomic and molecular characteristics, strains S88T and W115T represent two novel species of the genus Roseovarius, for which the names Roseovarius phycicola sp. nov. (S88T =KACC 23423T =JCM 36647T) and Roseovarius rhodophyticola sp. nov. (W115T =KACC 23690T =JCM 36651T) are proposed, respectively.

Aurantiacibacter flavus sp. nov. and Aurantiacibacter gilvus sp. nov., isolated from the mudflat of Suaeda japonica colonies.

Two novel strains were isolated from the mudflat of Suaeda japonica colonies in Incheon, Republic of Korea. Designated as DGU5T and DGU6T, these strains were Gram-stain-negative, facultatively anaerobic and rod-shaped and had yellowish colonies. Both strains were determined to belong to the genus Aurantiacibacter through phylogenetic analysis of their 16S rRNA sequences and draft genomes. The cells of strain DGU5T were non-motile and grew at temperatures ranging between 7-45°C (optimum, 25-30°C), pH 6.0-10.0 (optimum, 7.0-8.0) and in the presence of 0-11.0% NaCl (optimum, 2.0%). The cells of strain DGU6T were non-motile and grew in temperatures ranging from 10-45 °C (optimum, 30-35°C), pH 3.0-10.0 (optimum, 7.0-8.0) and in the presence of 0-11.0% NaCl (optimum, 2.0%). Overall genome relatedness index calculations revealed average nucleotide identity values (72.3-88.6%) and digital DNA-DNA hybridization values (18.8-35.9%) aligning with those of the genus Aurantiacibacter. The major fatty acids in both strains were C17:1 ω6c and summed feature 8 (C18:1 ω6c/C18:1 ω7c), while the predominant polar lipids were sphingoglycolipid, phosphatidylglycerol, and diphosphatidylglycerol. Phylogenetic, average nucleotide identity, digital DNA-DNA hybridization, physiological, and biochemical data collectively demonstrated the distinctiveness of the novel strains from other members within the family Erythrobacteraceae. We propose the names A. flavus sp. nov. (type strain DGU5T = KACC 23720T = TBRC 19015T) and A. gilvus sp. nov. (type strain DGU6T = KACC 23721T = TBRC 19016T) for the two strains.

Parasalinivibrio latis gen. nov., sp. nov., isolated from the distal gut of healthy farmed Asian Seabass (Lates calcarifer).

Asian Seabass (Lates calcarifer) is widely farmed as a sustainable source of protein for countries in the tropical Indo-West Pacific region. However, microbial species of the gut microbiome of healthy Asian Seabass remain largely uncharacterized and uncultured. Here, we analysed the microbial composition along the gastrointestinal tract of a farmed healthy Asian Seabass. We used different cultivation approaches to obtain isolates from the seabass intestinal tract and describe the isolation and characterization of a novel strain, TLL-SE01T. Analysis of the strain's 16S rRNA gene indicates that the strain belongs to the family Vibrionaceae with Photobacterium damselae as its closest relative, albeit sharing only 94.8% (aligned region 1553bp) nucleotide identity. Comparative genomic analysis with all validly published Vibrionaceae species with available genomes revealed average nucleotide identity (ANI) and DNA-DNA hybridisation (DDH) values of around 70% and 24% respectively to strain TLL-SE01T, which are well below proposed thresholds for species delineation (ANI, 95-96%; DDH, 70%). The alignment fraction and ANI genus demarcation boundaries for all genera in the Vibrionaceae family were determined for which strain TLL-SE01T is well below the calculated values, indicating that it belongs to a novel genus. Single- and core-gene phylogenetic analysis places strain TLL-SE01T in a monophyletic clade, further supporting its designation to a novel genus. Phenotypic comparison between strain TLL-SE01T and its close relatives indicated additional differences, such as growth response at different salt concentrations and different metabolic capabilities. Based on genotypic, phylogenetic and phenotypic differences to other Vibrionaceae species, we propose a novel species in a new genus, Parasalinivibrio latis gen. nov. sp. nov. and strain TLL-SE01T (= BCRC 81435T = JCM 36283T) as the type strain.

Microbispora maris sp. nov., a novel actinobacterium isolated from the gill of the leopard coral grouper (Plectropomus leopardus).

A novel actinomycete, designated strain ZYX-F-249T, was isolated from the gill of a leopard coral grouper in Yongxing Island, Hainan Province, China. Based on 16S rRNA gene sequence analysis, strain ZYX-F-249T belonged to the genus Microbispora, with high similarities to Microbispora rosea ATCC 12950T (98.7 %), Microbispora hainanensis 211020T (98.5 %), Microbispora clausenae CLES2T (98.4 %), Microbispora bryophytorum NEAU-TX2-2T (98.1 %) and Microbispora oryzaeRL4-1ST (98.0 %). Phylogenetic analysis of the 16S rRNA gene sequence of strain ZYX-F-249T showed that the strain formed a stable subclade with Microbispora rosea ATCC 12950T in the genus Microbispora. The cell wall of the novel isolate contained meso-diaminopimelic acid, while whole-cell sugars were madurose, glucose and ribose. The predominant menaquinones were MK-9(H6), MK-9(H4), MK-9(H2) and MK-9(H0). The characteristic phospholipids (PLs) were phosphatidylethanolamine, phosphatidylinositol, diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylinositol mannoside, glycophospholipids and an unknown PL. The major cellular fatty acids (>10%) were iso-C16 : 0 and iso-C18 : 0. Genome sequencing showed a DNA G+C content of 71.6%. The low average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity values demonstrated that strain ZYX-F-249T could be readily distinguished from closely related species. Based on data from these phylogenetic, chemotaxonomic and physiological characteristics, strain ZYX-F-249T represents a novel species of the genus Microbispora, for which the name Microbispora maris sp. nov. is proposed. The type strain is ZYX-F-249T (=CCTCC AA 2023030T =JCM 36778T).

Bifidobacterium favimelis sp. nov., isolated from black comb honey.

A strain of the bacterial genus Bifidobacterium, IMAU50987T, that was isolated from black comb honey from Lianghe County, Dehong Autonomous Prefecture, Yunnan Province, China, could not be accurately identified as a known species using 16S rRNA gene sequencing. In this study, the taxonomic status of strain IMAU50987T was evaluated using whole-genome sequencing combined with phenotypic analyses. Strain IMAU50987T was Gram-positive, non-motile and non-spore-forming. Comparative of 16S rRNA gene sequence similarity analysis indicated that strain IMAU50987T could be attributed to the genus Bifidobacterium and most closely related to the B. mellis Bin7NT (98.5% identity). Comparative sequence analysis of eight housekeeping genes (pyrG, thrS, glnA1, recA, tuf, dnaB, rpoC and xfp) showed that strain IMAU50987T was different from closely related species. Phylogenomic analysis based on the 88 single-copy genes in the phylogenetic tree showed that strain IMAU50987T belonged to the B. asteroides group. The highest average nucleotide identity value for strain IMAU50987T was 77.0% in comparison with B. choladohabitans B14384H11T and B. apis F753-1T. However, the highest digital DNA-DNA hybridization value was 23.1% compared with B. apis F753-1T and B. xylocopae XV2T; the highest average amino acid identity value was 73.2% compared with B. polysaccharolyticum W8117T. These values are below the accepted threshold for novel species boundaries. The draft genome size of strain IMAU50987T was 1.9 Mb, and the DNA G+C content was 63.5%. The major cellular fatty acids of strain IMAU50987T were C16 : 0, C18 : 1 ω9c and Sum in Feature 7 (C19 : 1 ω7c/C19 : 1 ω6c). The cell-wall peptidoglycan was of the A3α l-Lys-Gly type. Based on data from this study, a novel species, Bifidobacterium favimelis sp. nov., is proposed with the type strain IMAU50987T (= GDMCC 1.4189T = JCM 36315T).

Cultivation of deep-sea bacteria from the Northwest Pacific Ocean and characterization of Limnobacter profundi sp. nov., a phenol-degrading bacterium

Despite previous culture-independent studies highlighting the prevalence of the order Burkholderiales in deep-sea environments, the cultivation and characterization of deep-sea Burkholderiales have been infrequent. A total of 243 deep-sea bacterial strains were isolated from various depths in the Northwest Pacific Ocean, with 33 isolates (13.6%) from a depth of 4000 m classified into Burkholderiales. Herein, we report the isolation and genome characteristics of strain SAORIC-580T, from a depth of 4000 m in the Northwest Pacific Ocean. The strain showed a close phylogenetic relationship with Limnobacter thiooxidans CS-K2T, sharing 99.9% 16S rRNA gene sequence identity. The complete whole-genome sequence of strain SAORIC-580T comprised 3.3 Mbp with a DNA G+C content of 52.5%. Comparative genomic analysis revealed average nucleotide identities between 79.4–85.7% and digital DNA-DNA hybridization values of 19.9–29.5% when compared to other Limnobacter genomes, indicating that the strain represents a novel species within the genus. Genomic analysis revealed unique adaptations to deep-sea conditions, including genes associated with phenol degradation, stress responses, cold adaptation, heavy metal resistance, signal transduction, and carbohydrate metabolism. The SAORIC-580T genome was found to be more abundant in the deep sea than at the surface in the trenches of the Northwest Pacific Ocean, suggesting adaptations to the deep-sea environment. Phenotypic characterization highlighted distinct differences from other Limnobacter species, including variations in growth conditions, enzyme activities, and phenol degradation capabilities. Chemotaxonomic markers of the strain included ubiquinone-10, major fatty acids such as C16:0, C16:1, and C18:1, and major polar lipids including phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. Based on the polyphasic taxonomic data, it is concluded that strain SAORIC-580T (= KACC 21440T = NBRC 114111T) represents a novel species, for which the name Limnobacter profundi sp. nov. is proposed.

Blastococcus montanus sp. nov., a multi-stress-resistant and bacteriostatic-producing bacterium isolated from the Flaming Mountain, Xinjiang,China.

A bacterial strain designated HT6-4T was isolated from soil samples collected from the Flaming Mountain, Xinjiang, PR China. The purpose of this study was to describe a novel species and its characteristics, through genome sequencing and analysis of the relationship between the members of the genus Blastococcus, and explore the antiradiation, antioxidation and antibacterial capabilities of strain HT6-4T. The polyphasic study confirmed the affiliation of strain HT6-4T with the genus Blastococcus. Strain HT6-4T was aerobic, Gram-stain-positive, non-budding, non-motile, catalase-positive and oxidase-negative. It grew at 10-37 °C, pH 5.0-8.0 and 0-4% (w/v) NaCl. Colonies were circular, smooth and bright orange in colour. In addition, strain HT6-4T was drought tolerant. The predominant menaquinone was MK-9, with MK-8 as the minor component. The polar lipids of strain HT6-4T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phospholipids, an unidentified aminolipid and two unidentified phospholipids. Whole-cell hydrolysates contain meso-diaminopimelic acid as the diagnostic diamino acid and ribose and galactose as diagnostic sugars. Its major fatty acids were iso-C16 : 0, C17 : 1 ω8c and C18 : 1 ω9c. The genome of strain HT6-4T was 4.30 Mb in the whole-genome shotgun project. The G+C content was 73.9 mol%. The phylogenetic analysis based on the 16S rRNA gene sequence showed that strain HT6-4T was closely related to Blastococcus jejuensis KST3-10T(97.9%), Blastococcus capsensis BMG 804T(97.8%), Blastococcus aggregatus DSM 4725T(97.5%), Blastococcus saxobsidens BC 444T(97.5%), Blastococcus xanthinilyticus BMG 862T(97.5%) and Blastococcus litoris GP-S2-8T(97.5%). The average nucleotide identity (OrthoANI) and digital DNA-DNA hybridization (dDDH) values among strain HT6-4T and B. jejuensis KST3-10T, B. capsensis BMG 804T, B. aggregatus DSM 4725T, B. saxobsidens BC 444T, B. xanthinilyticus BMG 862T and B. litoris GP-S2-8T were below the species delimitation thresholds. The genome of strain HT6-4T contained antiradiation genes, antioxidant genes and antibacterial genes. Based on its morphological, physiological and chemical taxonomic characteristics, strain HT6-4T (=KCTC 59234T =GDMCC 1.4386T) should be classified as a novel species of the genus Blastococcus with the proposed name Blastococcus montanus sp. nov.