Haloterrigena Gelatinilytica sp. nov., A New Extremely Halophilic Archaeon Isolated From Salt-Lake

Two extremely halophilic strains, designated SYSU A558-1T and SYSU A121-1, were isolated from a saline sediment sample collected from Aiding salt lake, China. Cells of strains SYSU A558-1T and SYSU A121-1 were Gram-stain-negative, coccoid, and non-motile. The isolates were aerobic and grew at NaCl concentration of 10-30% (optimum, 20-22%), at 20-55ºC (optimum, 37-42ºC) and at pH 6.5-8.5 (optimum, 7.0-8.0). Cells lysed in distilled water. Major polar lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, disulphated diglycosyl diether-1 and one unidentified glycolipid. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that the two strains SYSU A558-1T and SYSU A121-1 were closely related to the membranes of the genus Haloterrigena. Phylogenetic trees based on strains SYSU A558-1T and SYSU A121-1 16S rRNA gene sequence, rpoB' gene sequence and concatenation of 87 protein markers demonstrated a robust clade with Haloterrigena turkmenica, Haloterrigena salifodinae and Haloterrigena salina. The genomic DNA G+C contents of strains SYSU A558-1T and SYSU A121-1 were 65.8 and 65.0%, respectively. Phenotypic, chemotaxonomic characteristics and phylogenetic properties suggested that the two strains SYSU A558-1T and SYSU A121-1 represent a novel species of the genus Haloterrigena, for which the name Haloterrigena gelatinilytica sp. nov. is proposed. The type strain is SYSU A558-1T (= KCTC 4259T = CGMCC 1.15953T).

Phylogeny of different Helicobacter pylori strains could not be explained by 16S rRNA gene due to high similarity in gene sequence across strains

Understanding the evolutionary relatedness of different strains of a species helped identify strain-specific differences that may be useful for disease diagnosis and treatment. Typically, such strain level typing would be augmented by molecular assays such as DNA sequencing, and phylogenetic tree analysis. This work utilizes public data on the 16S rRNA gene sequence of different strains of Helicobacter pylori to help plot the phylogenetic tree that describes the evolutionary trajectories of the different strains. Results from multiple sequence alignment reveals high level of conservation in 16S rRNA gene sequence across strains. This then translates into a phylogenetic tree structure that suggests very close evolutionary relationships of the different strains except for one outlier strain. Even in the case of the outlier strain, its evolutionary distance from other brethren was also not large. Overall, the results obtained in this study indicates that 16S rRNA gene may not capture strain-level phylogeny between different strains of the same species, and point to efforts in elucidating this phylogenetic effect in other genes of the species. Such genes may be involved in virulence during pathogenesis in humans, and may thus be subjected to higher evolutionary pressure and natural selection.

Enterococcus innesii sp. nov., isolated from the wax moth Galleria mellonella

Four bacterial strains were isolated from two different colony sources of the wax moth Galleria mellonella. They were characterized by a polyphasic approach including 16S rRNA gene sequence analysis, core-genome analysis, average nucleotide identity (ANI) analysis, digital DNA–DNA hybridization (dDDH), determination of G+C content, screening of antibiotic resistance genes, and various phenotypic analyses. Initial analysis of 16S rRNA gene sequence identities indicated that strain GAL7T was potentially very closely related to Enterococcus casseliflavus and Enterococcus gallinarum , having 99.5–99.9 % sequence similarity. However, further analysis of whole genome sequences revealed a genome size of 3.69 Mb, DNA G+C content of 42.35 mol%, and low dDDH and ANI values between the genomes of strain GAL7T and closest phylogenetic relative E. casseliflavus NBRC 100478T of 59.0 and 94.5 %, respectively, indicating identification of a putative new Enterococcus species. In addition, all novel strains encoded the atypical vancomycin-resistance gene vanC-4. Results of phylogenomic, physiological and phenotypic characterization confirmed that strain GAL7T represented a novel species within the genus Enterococcus , for which the name Enterococcus innesii sp. nov. is proposed. The type strain is GAL7T (=DSM 112306T=NCTC 14608T).

Streptomyces silvae sp. nov., isolated from forest soil

A bacterial strain, named For3T, was isolated from forest soil sampled in Champenoux, France. Based on its 16S rRNA gene sequence, the strain was affiliated to the family Streptomycetaceae and, more specifically, to the genus Streptomyces . The strain had 99.93% 16S rRNA gene sequence similarity to its closest relative strains Streptomyces pratensis ATCC 33331T, Streptomyces anulatus ATCC 27416T, Streptomyces setonii NRRL ISP-5322T and Kitasatospora papulosa NRRL B-16504T. The phylogenomic tree using the genome blast distance phylogeny method showed that the closest relative strain was Streptomyces atroolivaceus NRRL ISP-5137T and that For3T represents a new branch among the Streptomyces . Genome relatedness indexes revealed that the average nucleotide identity and digital DNA–DNA hybridization values between For3T and its closest phylogenomic relative ( S. atroolivaceus NRRL ISP-5137T) were 88.39 and 39.2 %, respectively. The G+C content of the genome was 71.4 mol% and its size was 7.96 Mb with 7492 protein-coding genes. Strain For3T harboured complete metabolic pathways absent in the closest relative strains such as cellulose biosynthesis, glycogen degradation I, glucosylglycerate biosynthesis I. Anteiso-C15:0, iso-C15:0, anteiso-C17:0 and MK-9(H4)/MK-9(H6) were the predominant cellular fatty acids and respiratory quinones, respectively. Phenotypic and genomic data supported the assignment of strain For3T to a novel species Streptomyces silvae sp. nov., within the genus Streptomyces , for which the type strain is For3T (=CIP 111908T=LMG 32186T).

Genome Mining Revealed Polyhydroxybutyrate Biosynthesis by Ramlibacter Agri Sp. Nov., Isolated from Agriculture Soil in Korea

A white-colony-forming, aerobic, motile and Gram-stain-negative bacterium, designated G-1-2-2T was isolated from soil of agriculture field near Kyonggi University, Republic of Korea. Strain G-1-2-2T synthesize the polyhydroxybutyrate and could grow at 10–35°C. The phylogenetic analysis of its 16S rRNA gene sequence, strain G-1-2-2T formed a lineage within the family Comamonadaceae and clustered as a member of the genus Ramlibacter. The 16S rRNA gene sequence of strain G-1-2-2T showed high sequence similarities with Ramlibacter ginsenosidimutans BXN5-27T (97.9%), Ramlibacter monticola G-3-2T (97.9%) and Ramlibacter alkalitolerans CJ661T (97.4%). The sole respiratory quinone was ubiquinone-8 (Q-8). The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, and an unidentified phospholipid. The principal cellular fatty acids were C16:0, cyclo-C17:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω6c). The genome of strain G-1-2-2T was 7,200,642 bp long with 13 contigs, 6,647 protein-coding genes, and DNA G+C content of 68.9%. The average nucleotide identity and in silico DNA–DNA hybridization values between strain G-1-2-2T and closest members were ≤81.2 and 24.1%, respectively. The genome of strain G-1-2-2T showed eight putative biosynthetic gene clusters responsible for various secondary metabolites. Genome mining revealed the presence of atoB, atoB2, phaS, phbB, phbC, bhbD genes in the genome which are responsible for polyhydroxybutyrate biosynthesis. Based on these data, strain G-1-2-2T represents a novel species in the genus Ramlibacter, for which the name Ramlibacter agri sp. nov. is proposed. The type strain is G-1-2-2T (= KACC 21616T = NBRC 114389T).

The effect of the use of commercial tempeh starter on the diversity of Rhizopus tempeh in Indonesia

At present, only a single Rhizopus species, R. microsporus, can be found in fresh tempeh produced in Java, Indonesia. The loss of diversity of Rhizopus in tempeh has been associated with the widespread use of commercial tempeh starter in Indonesia since the 2000s. However, the identities of the previous Rhizopus strains associated with tempeh, which have been preserved in a culture collection in Indonesia, have not been verified. The present study aimed to verify the identities of 22 Rhizopus strains isolated from tempeh produced using the traditional tempeh starters from the 1960s to the 2000s. Phylogenetic analysis based on the ITS regions in the rRNA gene sequence data, revealed that the Rhizopus strains belonged to the species R. arrhizus (five strains); R. delemar (14 strains); and R. microsporus (three strains). Verification of the identities of these Rhizopus strains in the present study confirmed the loss of diversity of Rhizopus species in tempeh produced in Indonesia, particularly in Java. Our findings confirmed that the morphological changes in Rhizopus species isolated from tempeh as a result of centuries of domestication.

Devosia litorisediminis sp. nov., isolated from a sand dune

A Gram-negative, aerobic, non-motile, and rod-shaped bacterial strain, designated BSSL-BM10T, was isolated from a sand dune that was collected from the Yellow Sea, Republic of Korea. It was subjected to a polyphasic taxonomic study. 16S rRNA gene sequence analysis showed that strain BSSL-BM10T fell phylogenetically within the radiation comprising type strains of Devosia species. The 16S rRNA gene sequence of strain BSSL-BM10T shared sequence similarities of 98.2% with the type strain of D. naphthalenivorans and 93.5-97.7% with type strains of other Devosia species. ANI and dDDH values between strain BSSL-BM10T and type strains of 18 Devosia species were 71.0-78.4% and 18.8-21.5%, respectively. The DNA G+C content of strain BSSL-BM10T was 60.9% based on its genomic sequence data. Strain BSSL-BM10T contained Q-10 as the predominant ubiquinone and 11-methyl C18:1 ω7c, C18:1 ω7c, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), and C16:0 as its major fatty acids. Major polar lipids of strain BSSL-BM10T were phosphatidylglycerol and two unidentified glycolipids. Strain BSSL-BM10T showed distinguished phenotypic properties with its phylogenetic and genetic distinctiveness separated from recognized Devosia species. Based on data presented in this study, strain BSSL-BM10T should be placed in the genus Devosia. The name Devosia litorisediminis sp. nov. is proposed for strain BSSL-BM10T (= KACC 21633T = NBRC 115152T).

Devosia equisanguinis sp. nov., isolated from horse blood

A Gram-stain-negative, aerobic, non-endospore-forming organism isolated from horse blood was studied for its taxonomic allocation. On the basis of 16S rRNA gene sequence similarity comparisons, strain M6-77T grouped within the genus Devosia and was most closely related to Devosia elaeis (97.6 %) and Devosia indica (97.55 %). The 16S rRNA gene sequence similarity to type strains of other Devosia species was below 97.5 %. The average nucleotide identity and digital DNA–DNA hybridization values between the M6-77T genome assembly and those of the closest relative Devosia type strains were <85 and <25 %, respectively. Strain M6-77T grew optimally at 25–37 °C (range: 10–36 °C), at a pH range of pH 6.5–10.5 and in the presence of up to 3 % (w/v) NaCl. The fatty acid profile from whole-cell hydrolysates supported the allocation of the strain to the genus Devosia . Major fatty acids were C18 : 1 ω7c, 11-methyl C18 : 1 ω7c and C16 : 0. The quinone system consisted exclusively of ubiquinone Q-10. The polar lipid profile was composed of the major lipids diphosphatidylglycerol, phosphatidylglycerol and three unidentified glycolipids. In the polyamine pattern, putrescine was predominant and spermidine was detected in moderate amounts. The diamino acid of the peptidoglycan was meso-diaminopimelic acid. In addition, the results of physiological and biochemical tests also allowed phenotypic differentiation of strain M6-77T from the closely related species. Hence, M6-77T represents a new species of the genus Devosia , for which we propose the name Devosia equisanguinis sp. nov., with M6-77T (=CIP 111628T=LMG 30659T=CCM 8868T) as the type strain.

Sphingobacterium hungaricum sp. nov. a novel species on the borderline of the genus Sphingobacterium

A Gram-reaction-negative bacterial strain, designated Kb22T, was isolated from agricultural soil and characterized using a polyphasic approach to determine its taxonomic position. On the basis of 16S rRNA gene sequence analysis, the strain shows highest similarity (94.39 %) to Sphingobacterium nematocida M-SX103T. The highest average nucleotide identity value (71.83 %) was found with Sphingobacterium composti T5-12T, and the highest amino acid identity value (66.65 %) was found with Sphingobacterium olei HAL-9T. Cells are aerobic, non-motile rods. The isolate was found to be positive for catalase and oxidase tests. The assembled genome of strain Kb22T has a total length of 4,06 Mb, the DNA G+C content is 38.1 mol%. The only isoprenoid quinone is menaquinone 7 (MK-7). The major fatty acids are iso-C15:0 (28.4%), summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH) (25.7 %) and iso-C17:0 3-OH (19.7 %). Based on phenotypic characteristics and phylogenetic results, it is concluded that strain Kb22T is a member of the genus Sphingobacterium and represents a novel species for which the name Sphingobacterium hungaricum sp. nov. is proposed. The type strain of the species is strain Kb22T (=LMG 31574T=NCAIM B.02638T).