Comparative pangenome analysis of Enterococcus faecium and Enterococcus lactis provides new insights into the adaptive evolution by horizontal gene acquisitions.

BACKGROUND: Enterococcus faecium and E. lactis are phylogenetically closely related lactic acid bacteria that are ubiquitous in nature and are known to be beneficial or pathogenic. Despite their considerable industrial and clinical importance, comprehensive studies on their evolutionary relationships and genomic, metabolic, and pathogenic traits are still lacking. Therefore, we conducted comparative pangenome analyses using all available dereplicated genomes of these species. RESULTS: E. faecium was divided into two subclades: subclade I, comprising strains derived from humans, animals, and food, and the more recent phylogenetic subclade II, consisting exclusively of human-derived strains. In contrast, E. lactis strains, isolated from diverse sources including foods, humans, animals, and the environment, did not display distinct clustering based on their isolation sources. Despite having similar metabolic features, noticeable genomic differences were observed between E. faecium subclades I and II, as well as E. lactis. Notably, E. faecium subclade II strains exhibited significantly larger genome sizes and higher gene counts compared to both E. faecium subclade I and E. lactis strains. Furthermore, they carried a higher abundance of antibiotic resistance, virulence, bacteriocin, and mobile element genes. Phylogenetic analysis of antibiotic resistance and virulence genes suggests that E. faecium subclade II strains likely acquired these genes through horizontal gene transfer, facilitating their effective adaptation in response to antibiotic use in humans. CONCLUSIONS: Our study offers valuable insights into the adaptive evolution of E. faecium strains, enabling their survival as pathogens in the human environment through horizontal gene acquisitions.

Preliminary Study on Effect of Lactiplantibacillus plantarum on Osteoporosis in the Ovariectomized Rat.

Osteoporosis is a growing global health concern primarily associated with decreased estrogen in postmenopausal women. Recently, some strains of probiotics were examined for potential anti-osteoporotic effects. This study intended to evaluate the impacts of Lactiplantibacillus plantarum MGE 3038 strain (MGE 3038) in ovariectomized rats. For this purpose, twelve weeks old female Wistar rats (n=21; 250-300 g) were divided into 3 groups; ovariectomy (OVX) group, OVX/MGE 3038 group and Sham group (control). In these groups; two went through respective OVX and one had daily MGE 3038 administration through oral gavage. Prior to 16 weeks after OVX, we collected blood samples and extracted the tibiae. We scanned the extracted tibiae by in-vivo micro-computed tomography (micro-CT) and evaluated pathology by hematoxylin and eosin (H&E) and Masson's trichrome staining. The serum levels of C-telopeptide of type I collagen (CTX), osteocalcin (OC), and the receptor activator of nuclear factor-ĸB ligand (RANKL) were examined. The OVX/MGE 3038 group showed increases in bone mineral density, trabecular bone volume, trabecular number, and trabecular thickness (Tb.Th), and a decrease in trabecular spacing than the OVX group. However, OVX/MGE 3038 group and control group were measurably comparable in Tb.Th. Micro-CT, H&E, and Masson's trichrome findings exhibited increased preservation and maintenance of trabecular bone structure in the OVX/MGE 3038 group in comparison to the OVX group. In serum, the levels of CTX, OC and RANKL were significantly different between the OVX and OVX/MGE 3038 groups. Taken together, L. plantarum MGE 3038 could be helpful for the treatment of osteoporosis.

Cutibacterium acnes KCTC 3314 Growth Reduction with the Combined Use of Bacteriophage PAP 1-1 and Nisin.

Severe acne has high psychological impacts recorded worldwide, from depression to suicide. To control acne infection, bacteriophage could be used in synergy or combination with antibiotics/antimicrobials. Bacteriophages are specific to their hosts without interfering with the normal skin microbes and have the ability to lyse bacterial cells. In this current study, the bacteriophage PAP 1-1 was isolated, characterized, and tested against the pathogenic acne-causing bacterium Cutibacterium acnes. Examination under transmission electron microscopy (TEM) revealed that the newly isolated phage has a morphology typical of siphoviruses. Phylogenetic analysis, utilizing the maximum likelihood (ML) algorithm based on complete genome sequences, revealed that PAP 1-1 clustered together with bacteriophages active to Propionibacterium acnes (now known as C. acnes), forming a distinct evolutionary lineage. The genomic analysis further identified the presence of an endolysin gene in PAP 1-1, suggesting its potential to regulate the growth of C. acnes. Subsequent experiments conducted in RCM broth confirmed the ability of PAP 1-1 to effectively control the proliferation of C. acnes. In combination with bacteriocin from Lactococcus lactis CJNU 3001 and nisin, PAP 1-1 greatly decreased the viable cell counts of C. acnes in the broth.

Genome-wide transcriptional response of Escherichia coli O157:H7 to light-emitting diodes with various wavelengths.

We investigated the physiological and transcriptomic response of Escherichia coli at the early stationary phase to light-emitting diodes with different wavelengths. The growth and metabolic changes of E. coli O157:H7 were examined under the influence of 465, 520, and 625 nm illuminated light. Under 465 nm illumination, the growth of E. coli O157:H7 was significantly retarded compared to 520 nm and 625 nm illumination and non-illuminated control. Metabolic changes were examined under these illumination and non-illuminated conditions based on transcriptomic reads. Transcriptomic response under 520 nm and 625 nm remained almost similar to control except few up-and down-regulated genes. Carbohydrates metabolic transcriptomic reads were greatly down-regulated under 465 nm illumination compared to 520 nm and 625 nm illumination and non-illuminated control showing depletion of glucose as a sole energy source during the exponential phase. Fatty acid degradation such as fad regulon-related genes was up-regulated in cells under 465 nm illumination revealing the shifting of cells to use fatty acid as a new carbon energy source during the early stationary phase. Exposure of E. coli O157:H7 cells to 465 nm illuminated light down-regulated virulence factor genes such as hlyA, hlyB, hlyC, stx1A, stx2B, paa, and bdm. Under the stress of 465 nm illumination, expression of stress and flagellar motility-related genes were up-regulated causing consumption of energy and reduction in cell growth. Also, oxidative phosphorylated transcriptomic reads were up-regulated under 465 nm illumination probably due to the production of ROS that might involve in the reduction of cell growth during the early stationary phase. These results indicate that pathogenic E. coli O157:H7 respond differentially to a different wavelength of the light-emitting diodes used in this study.

Combination Treatment for Inhibition of the Growth of Staphylococcus aureus with Recombinant SAP8 Endolysin and Nisin.

Staphylococcus aureus, a pathogenic species of genus Staphylococcus involved in foodborne illness always remain among the top priorities of the world major concerns. In the present study, we have used recombinant SAP8 endolysin from the bacteriophage SAP8 and commercial nisin to inhibit the viability of pathogenic S. aureus KCTC 3881 cells; however, the approach was not identified as cost-effective. A gradual decrease in the viable S. aureus KCTC 3881 cell counts was observed with an increase in the concentrations of recombinant SAP8 endolysin and nisin. However, combined treatment with recombinant SAP8 endolysin and nisin decreased the viable S. aureus KCTC 3881 cell counts in a significant manner. The combination of 0.01 µM of recombinant SAP8 endolysin with 9 IU/mL and 18 IU/mL of nisin demonstrated a promising decrease in the viable cell counts of the strain. Under the scanning electron microscope, the combination treatment with 0.01 µM of recombinant SAP8 endolysin and 18 IU/mL of nisin showed complete cellular destruction of S. aureus KCTC 3881. We propose that a combination of recombinant SAP8 endolysin and nisin could be a strong alternative to antibiotics to control the growth of S. aureus including MRSA.

Arsenicicoccus cauae sp. nov., isolated from the blood of a pediatric gastroenteritis patient.

A Gram-stain-positive coccus was isolated from the blood of a paediatric patient suffering from gastroenteritis. The taxonomic position of this catalase-positive, non-motile, non-spore-forming facultative anaerobe designated as strain MKL-02T was investigated using a polyphasic approach. Colonies grown on tryptic soy agar with 10 % sheep blood were circular, creamy yellow, and convex. Phylogenetic analysis based on 16S rRNA gene and whole-genome sequences revealed that this strain was most closely related to Arsenicicoccus bolidensis CCUG 47306T within the cluster of the genus Arsenicicoccus. Average nucleotide identity and digital DNA-DNA hybridization values between strain MKL-02T and A. bolidensis DSM 15745T, A. dermatophillus DSM 25571T and A. piscis DSM 22760T were 89.5 and 37.0 %, 79.6 and 22.4 %, and 75.9 and 21.0 %, respectively. The genomic size of strain MKL-02T was 3 423 857 bp with a 72.7 mol% G+C content. Growth was observed at 10-45 °C (optimum, 37-40 °C) and pH 6.0-10.0 (optimum, pH 7.0), in the presence of 0-10 % (w/v) NaCl (optimum, 0.5 %). Cells of strain MKL-02T were non-motile cocci and 0.50-0.60 µm long, as determined by transmission electron microscopy. The strain was catalase-positive and oxidase-negative. The major fatty acid type (>10 % of total) was C15 : 0. The polar lipid profile consisted of two unidentified phospholipids, three unidentified lipids and an unidentified aminophospholipid. The strain contained MK-8 (H4) as the predominant menaquinone. Based on phylogenetic and phenotypic considerations, it is proposed that strain MKL-02T be classified as a new species, named Arsenicicoccus cauae sp. nov. The type strain is MKL-02T (=NCCP 16967T=JCM 34624T).

Ramlibacter terrae sp. nov. and Ramlibacter montanisoli sp. nov., Isolated from Soil.

Two gram-negative, catalase-positive, strictly aerobic, and white colony-forming bacteria, strains H242T and B156T, were isolated from soil in South Korea. Cells of strain H242T were oxidase-positive and non-motile short rods, while those of strain B156T were oxidase-negative and long non-motile rods. Ubiquinone-8 was identified as the sole isoprenoid quinone in both strains. C16:0, cyclo-C17:0, and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol were identified in both strains as the major cellular fatty acids and polar lipids, respectively. The DNA G+C contents of strains H242T and B156T were 69.4 mol% and 69.3 mol%, respectively. Phylogenetic analyses based on 16S rRNA and 92 concatenated core gene sequences revealed that strains H242T and B156T formed distinct phylogenic lineages from other Ramlibacter type strains. The DNA-DNA hybridization (DDH) value between strains H242T and B156T was 24.6%. Strains H242T and B156T were most closely related to Ramlibacter ginsenosidimutans BXN5-27T and Ramlibacter monticola G-3-2T with 98.4% and 98.6% 16S rRNA gene sequence similarities, respectively. Digital DDH values between strain H242T and R. ginsenosidimutans and between strain B156T and R. monticola were 23.5% and 26.1%, respectively. Phenotypic, chemotaxonomic, and molecular analyses indicated that strains H242T and B156T represent two novel species of the genus Ramlibacter, for which the names Ramlibacter terrae sp. nov. and Ramlibacter montanisoli sp. nov., respectively, are proposed. The type strains of R. terrae and R. montanisoli are H242T (=KACC 21667 T =JCM 33922T) and B156T (=KACC 21665 T =JCM 33920T), respectively.

Halomonas azerica sp. nov., Isolated from Urmia Lake in Iran.

A novel moderately halophilic, Gram-staining negative and facultative aerobic bacterium, designated as TBZ9T, was isolated from water of Urmia Lake in Azerbaijan region of Iran. The cells were found to be rod-shaped and motile, growing in the form of creamy, convex and shiny colonies. The strain could grow in the presence of NaCl at concentrations 1-17% (w/v) (optimum, 3%), temperatures 10-40 °C (optimum, 30 °C) and pH 6.0-11.0 (optimum, pH 7.0) on marine agar. Strain TBZ9T 16S rRNA gene sequence was related to the genus Halomonas showing highest similarities to Halomonas arcis AJ282T (98.4%), Halomonas songnenensis NEAU-ST10-39T (98.0%) and Halomonas lutescens Q1UT (97.8%). In the phylogenetic trees, strain TBZ9T formed a distinct branch closely related to a subclade inside the Halomonas genus. Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strain TBZ9T and H. arcis AJ282T (20.0%, 74.0%) and H. songnenensis NEAU-ST10-39T (19.8%, 75.2%) indicated that TBZ9T represents a distinct species. Evaluation of fatty acid contents determined C10:0, C16:0, C12:0 3-OH, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c) as major cellular fatty acids. The major quinone of strain TBZ9T was Q-9. Polar lipid patterns consisted of phosphatidylglycerol (PG), phosphatidylethanolamine (PE), two unidentified phospholipids (PL) and four unidentified polar lipids (L). The average DNA G + C content of strain TBZ9T is 55.4 mol%. Results from phenotypic, chemotaxonomic and molecular analysis suggest that the strain TBZ9T represents a novel species within the genus Halomonas for which the name Halomonas azerica sp. nov. is proposed. The type strain is TBZ9T (= KACC 21783T = LMG 25775T).

Lysobacter arenosi sp. nov. and Lysobacter solisilvae sp. nov. isolated from soil.

Two Gram-stain negative, yellow-pigmented, and mesophilic bacteria, designated strains R7T and R19T, were isolated from sandy and forest soil, South Korea, respectively. Both strains were non-motile rods showing catalase- and oxidase-positive activities. Both strains were shown to grow at 10-37°C and pH 6.0-9.0, and in the presence of 0-1.5% (w/v) NaCl. Strain R7T contained iso-C14:0, iso-C15:0, iso-C16:0, and summed feature 9 (comprising C16:0 10-methyl and/or iso-C17:1ω9c), whereas strain R19T contained iso-C11:0 3-OH, C16:1ω7c alcohol, iso-C11:0, iso-C15:0, iso-C16:0, and summed feature 9 (comprising C16:0 10-methyl and/or iso-C17:1ω9c) as major cellular fatty acids (> 5%). Both strains contained ubiquin-one-8 as the sole isoprenoid quinone and phosphatidylglycerol, phosphatidylethanolamine, and an unidentified phospholipid as the major polar lipids. The DNA G + C contents of strains R7T and R19T calculated from their genomes were 66.9 mol% and 68.9 mol%, respectively. Strains R7T and R19T were most closely related to Lysobacter panacisoli C8-1T and Lysobacter niabensis GH34-4T with 98.7% and 97.8% 16S rRNA sequence similarities, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that strains R7T and R19T formed distinct phylogenetic lineages within the genus Lysobacter. Based on phenotypic, chemotaxonomic, and molecular features, strains R7T and R19T represent novel species of the genus Lysobacter, for which the names Lysobacter arenosi sp. nov. and Lysobacter solisilvae sp. nov. are proposed. The type strains of L. arenosi and L. solisilvae are R7T (= KACC 21663T = JCM 34257T) and R19T (= KACC 21767T = JCM 34258T), respectively.

Devosia aurantiaca sp. nov., Isolated from Mountain Soil and Proposal of Albitalea gen. nov. to Replace the Illegitimate Prokaryotic Genus Name Geomonas Khan et al. 2020.

Strain H239T, a gram-negative, strictly aerobic, and oxidase-positive, and catalase-negative bacterium, was isolated from mountain soil in Gangwon-do of South Korea. Colonies were orange colored, and cells were motile rods with a single polar flagellum. Growth was observed between 25 and 30 °C (optimum, 30 °C), between pH 7.0 and pH 9.0 (optimum, pH 7.5), and in the presence of 0-1.5% (w/v) NaCl (optimum 0.5-1%). Ubiquinone-10 was detected as the sole respiratory quinone. The major fatty acids (>10%) of strain H239T were C18:1 ω7c, C18:0, and C16:0. The polar lipids detected from strain H239T consisted of two unidentified glycolipids, two unidentified phospholipids, and three unidentified polar lipids. The G+C content of strain H239T based on its genome sequence was 62.0 mol%. Comparative 16S rRNA gene sequence analysis indicated that strain H239T was most closely related to Devosia chinhatensis IPL18T (97.7%), Devosia submarina KMM 9415T (97.7%), and Devosia yakushimensis Yak96BT (97.3%). Phylogenetic analyses based on the 16S rRNA gene and whole-genome sequences revealed that strain H239T formed a distinct phyletic lineage as a new species within the genus Devosia. Based on its physiological, chemotaxonomic, and molecular properties, strain H239T represents a novel species of the genus Devosia, for which the name Devosia aurantiaca sp. nov. is proposed. The type strain is H239T (=KACC 21662T=JCM 33930T). In addition, because the prokaryotic genus name Geomonas Khan et al. 2020 is a later homonym of Geomonas Xu et al. 2020, the name is illegitimate (Principle 6 in the International Code of Nomenclature of Prokaryotes). Therefore, we propose to replace the problematic prokaryotic names Geomonas and Geomonas soli with Albitalea with Albitalea terrae, respectively.

Pseudoprevotella muciniphila gen. nov., sp. nov., a mucin-degrading bacterium attached to the bovine rumen epithelium.

A Gram-negative, strictly anaerobic mucin-degrading bacterium, which we designated strain E39T, was isolated from the rumen epithelium of Korean cattle. The cells were non-motile and had a coccus morphology. Growth of strain E39T was observed at 30-45°C (optimum, 39°C), pH 6.5-8.5 (optimum, pH 7.5), and in the presence of 0.0-1.0% (w/v) NaCl (optimum, 0.0-0.5%). Strain E39T contained C16:0, C18:0, C18:1 ω9c, iso-C15:0, and anteiso-C15:0 as the major fatty acids. The major polar lipids were phosphatidylethanolamine, unidentified aminophospholipid, and unidentified lipids. The major respiratory isoprenoid quinones were MK-8 and MK-9. The major fermented end-products of mucin were acetate and succinate. The G+C content of the genomic DNA was 46.4 mol%. Strain E39T was most closely related to Alloprevotella rava 81/4-12T with an 87.3% 16S rRNA gene sequence similarity. On the basis of phenotypic, chemotaxonomic, and molecular properties, strain E39T represents a novel genus of the family Prevotellaceae; as such, the name Pseudoprevotella muciniphila gen. nov., sp. nov. is proposed. A functional annotation of the whole genome sequences of P. muciniphila E39T revealed that this bacterium has a putative mucin-degrading pathway and biosynthetic pathways of extracellular polymeric substances and virulence factors which enable bacteria to adhere to the epithelial cells and avoid the host's immune responses.

Cupriavidus cauae sp. nov., isolated from blood of an immunocompromised patient.

A novel Gram-stain-negative, facultative aerobic and rod-shaped bacterium, designated as MKL-01T and isolated from the blood of immunocompromised patient, was genotypically and phenotypically characterized. The colonies were found to be creamy yellow and convex. Phylogenetic analysis based on 16S rRNA gene and whole-genome sequences revealed that strain MKL-01T was most closely related to Cupriavidus gilardii LMG 5886T, present within a large cluster in the genus Cupriavidus. The genome sequence of strain MKL-01T showed the highest average nucleotide identity value of 92.1 % and digital DNA-DNA hybridization value of 44.8 % with the closely related species C. gilardii LMG 5886T. The genome size of the isolate was 5 750 268 bp, with a G+C content of 67.87 mol%. The strain could grow at 10-45 °C (optimum, 37-40 °C), in the presence of 0-10 % (w/v) NaCl (optimum, 0.5%) and at pH 6.0-10.0 (optimum, pH 7.0). Strain MKL-01T was positive for catalase and negative for oxidase. The major fatty acids were C16 : 0, summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c and/or C16 : 1 ω6c/C16 : 1 ω7c) and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c). The polar lipid profile consisted of phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids and one unidentified polar lipid. Moreover, strain MKL-01T contained ubiquinone Q-8 as the sole respiratory quinone. Based on its molecular, phenotypic and chemotaxonomic properties, strain MKL-01T represents a novel species of the genus Cupriavidus; the name Cupriavidus cauae sp. nov. is proposed for this strain. The type strain is MKL-01T.

Maritimibacter harenae sp. nov. and Sneathiella litorea sp. nov.: members of Alphaproteobacteria isolated from sea sand.

Two Gram-negative, rod bacteria, strains DP07T and DP05T, showing catalase- and oxidase-positive activities, were isolated from sea sand in South Korea. Strain DP07T was strictly aerobic and had a yellow colony colour. Contrastingly, strain DP05T was facultatively aerobic and had a creamy colony colour. Both strains contained ubiquinone-10 as the sole isoprenoid quinone. Strain DP07T contained cyclo-C19:0 ω8c, C16:0, summed feature 8 (comprising C18:1 ω7c and/or C18:1 ω6c), and C16:0 2-OH as the major fatty acids and phosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid, and an unidentified polar lipid as the major polar lipids. Strain DP05T contained C16:0, cyclo-C19:0, and summed feature 8 (comprising C18:1 ω7c and/or C18:1 ω6c) as the major fatty acids and phosphatidylglycerol, phosphatidylethanolamine, an unidentified aminophospholipid, and two unidentified polar lipids as the major polar lipids. Cadaverine was detected as the predominant polyamine in both strains. The DNA G + C contents of strains DP07T and DP05T were 66.1 mol% and 51.8 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains DP07T and DP05T formed close phylogenetic lineages with Maritimibacter alkaliphilus HTCC2654T (similarity, 98.1%) and Sneathiella chungangensis CAU 1294 T (similarity, 98.5%), respectively. The average nucleotide identity and DNA-DNA hybridisation values between strain DP07T and M. alkaliphilus were 80.0% and 23.0%, respectively, and those between strain DP05T and S. chungangensis were 77.1% and 19.9%, respectively. Based on the results of phenotypic, chemotaxonomic, and molecular analyses, strains DP07T and DP05T represent two novel species of the genera Maritimibacter and Sneathiella, respectively, for which we propose the following names: Maritimibacter harenae sp. nov. and Sneathiella litorea sp. nov. The type strains of M. harenae and S. litorea were DP07T (= KACC 21429 T = JCM 33811 T) and DP05T (= KACC 21431 T = JCM 33810 T), respectively.

Paenibacillus silvestris sp. nov., Isolated from Forest Soil.

A bacterial strain, designated 5J-6T, was isolated from soil in Cheongnyeongpo, Republic of Korea. Cells were Gram-stain-positive, strictly aerobic, and motile rods and their catalase and oxidase activities were positive. Strain 5J-6T grew at 10-30 °C, pH 6.0-9.0, and 0-0.8% (w/v) NaCl concentration, with optimum growth at 25 °C, pH 6.5, and 0.4% NaCl concentration. Anteiso-C15:0 and iso-C16:0 were detected as the predominant fatty acids and menaquinone-7 was the sole isoprenoid quinone detected. Strain 5J-6T contained phosphatidylethanolamine, phosphatidylglycerol and an unidentified phospholipid as major polar lipids. The peptidoglycan belonged to the type A1γ meso-diaminopimelic acid. The G+C content of the genomic DNA calculated from the whole genomic sequence was 46.1 mol%. Phylogenetic analysis of strain 5J-6T based on 16S rRNA gene sequences placed the isolate into a member of the genus Paenibacillus. Sequence similarity analysis of 16S rRNA gene sequences revealed that strain 5J-6T was most closely related to Paenibacillus aceris KUDC4121T and Paenibacillus chondroitinus DSM 5051T with 98.76% and 98.42% similarities, respectively. Average nucleotide identity and in silico DNA-DNA hybridization values between strain 5J-6T and the type strain of P. aceris were 83.97% and 28.60%, respectively. Based on the phylogenetic and phenotypic characteristics and genomic data, strain 5J-6T could be considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus silvestris sp. nov. is proposed. The type strain is 5J-6T (= KACC 21430T = JCM 33812T).

Roseomonas algicola sp. nov., isolated from a green alga, Pediastrum duplex.

A Gram-stain-negative, strictly aerobic bacterium, designated strain PeD5T, was isolated from a green alga Pediastrum duplex from the Nakdong river of the Republic of Korea. Cells were non-motile cocci, catalase-negative and oxidase-positive. Growth of PeD5T was observed at 25-40 °C (optimum, 35 °C) and pH 5.0-10.0 (optimum, pH 7-8), and in the presence of 0-0.25% (w/v) NaCl (optimum, 0%). PeD5T contained C16:0, C18:1ω7c 11-methyl, summed feature 3 (comprising C16:1ω7c and/or C16:1ω6c) and summed feature 8 (comprising C18:1ω7c and/or C18:1ω6c) as major cellular fatty acids (>5%) and ubiquinone-10 as the sole isoprenoid quinone. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified phospholipid and an unidentified aminolipid were detected as major polar lipids. The genomic DNA G+C content of PeD5T was 71.0 mol%. PeD5T was most closely related to Roseomonas stagni HS-69T with a 97.6% 16S rRNA sequence similarity and shared less than 96.3% 16S rRNA sequence similarities with type strains of other species. Phylogenetic analysis based on 16S rRNA gene sequences indicated that PeD5T formed a phyletic lineage with Roseomonas stagni HS-69T within the genus Roseomonas. On the basis of results of phenotypic, chemotaxonomic and molecular analysis, strain PeD5T clearly represents a novel species of the genus Roseomonas, for which the name Roseomonas algicola sp. nov. is proposed. The type strain is PeD5T (=KACC 19925T=JCM 33309T).

Mucilaginibacter agri sp. nov. and Mucilaginibacter humi sp. nov., isolated from soil.

Two Gram-stain-negative, facultative anaerobic and non-motile bacteria, strains R11T and S1162T, were isolated from soil in the Republic of Korea. Both strains were catalase- and oxidase-positive and contained menaquinone-7 as the major isoprenoid quinone. Strain R11T contained summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), iso-C15:0, C16:0 and iso-C17:0 3-OH as major fatty acids and phosphatidylethanolamine, an unidentified aminophospholipid and an unidentified aminolipid as major polar lipids; while strain S1162T contained summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), iso-C15:0, iso-C17:0 3-OH, C16:0 and summed feature 9 (10-methyl C16:0 and/or iso-C17:1 ω9c) as major fatty acids and phosphatidylethanolamine and an unidentified aminophospholipid as major polar lipids. The DNA G+C contents of strains R11T and S1162T calculated from their whole genomes were 42.7 and 42.9 mol%, respectively. Results of phylogenetic analysis based on 16S rRNA gene sequences showed that strain R11T formed a phylogenetic lineage with Mucilaginibacter jinjuensis YC7004T and strain S1162T formed a distinct phyletic lineage from closely related type strains within the genus Mucilaginibacter. Strains R11T and S1162T were most closely related to M. jinjuensis YC7004T and Mucilaginibacter panaciglaebae BXN5-31T with 97.78 and 97.23% 16S rRNA gene sequence similarities, respectively. On the basis of phenotypic, chemotaxonomic and molecular analysis, strains R11T and S1162T represent two novel species of the genus Mucilaginibacter, for which the names Mucilaginibacter agri sp. nov. and Mucilaginibacter humi sp. nov. are proposed, respectively. The type strains of M. agri and M. humi are R11T (=KACC 21228T=JCM 33472T) and S1162T (=KACC 21669T=JCM 33916T), respectively.

Flavihumibacter soli sp. nov., Isolated from Soil.

A novel Gram-stain-positive, strictly aerobic, non-motile, rod-shaped bacterium, designated strain R14T was isolated from steep grove soil and taxonomically characterized using a polyphasic approach. Cells showed oxidase-negative and catalase-positive activities and grew at 25-37 °C (optimum, 30 °C), pH 5.0-8.0 (optimum, pH 6.5), and in the presence of 0-0.5% (w/v) NaCl (optimum, 0%). Phylogenetic trees based on 16S rRNA gene sequences revealed that strain R14T formed a phylogenetic lineage with Flavihumibacter genus members within the family Chitinophagaceae. Comparison of 16S rRNA gene sequences showed that strain R14T shared highest similarities with Flavihumibacter solisilvae 3-3T (97.5%), Flavihumibacter petaseus NBRC 106054T (96.7%), and Flavihumibacter stibioxidans YS-17T (96.1%). The G+C content of strain R14T calculated from its whole genome sequence was 44.6 mol%. Strain R14T contained menaquinone-7 and iso-C15:0, iso-C17:0 3-OH, C16:0, C15:1 ω6c, and iso-C15:0-G as the sole respiratory isoprenoid quinone and major cellular fatty acids, respectively. Major polar lipids consisted of phosphatidylethanolamine, two unidentified aminolipids, and four unidentified lipids. Based on phylogenetic, phenotypic, and chemotaxonomic characteristics of strain R14T, it could be concluded that strain R14T represents a novel species of the genus Flavihumibacter, for which the name Flavihumibacter soli sp. nov. is proposed. The type strain is R14T (=KACC 21229T = JCM 33473T).

Aestuariirhabdus litorea gen. nov., sp. nov., isolated from a sea tidal flat and proposal of Aestuariirhabdaceae fam. nov.

A Gram-negative, moderately halophilic and facultatively aerobic bacterium, designated strain GTF13T, was isolated from a sea tidal flat. Cells were curved rods and motile by a single polar flagellum showing catalase and oxidase activities. Growth was observed at 20-37 °C, pH 5.0-8.5 and 1.0-6.0 % (w/v) NaCl. Strain GTF13T contained C16:0, summed feature 3 (comprising C16 : 1 ω6c/C16 : 1 ω7c), summed feature 8 (comprising C18 : 1 ω6c/C18 : 1 ω7c) and C12 : 0 3-OH as major fatty acids and ubiquinone-9 and ubiquinone-8 as major quinones. Phosphatidylethanolamine and two unidentified phospholipids were detected as major polar lipids. The G+C content of the genomic DNA was 59.8 mol%. Strain GTF13T was most closely related to Simiduia agarivorans SA1T, Endozoicomonas montiporae CL-33T and Pseudomonas segetis FR1439T, belonging to different families or orders of the class Gammaproteobacteria, with less than 92.0 % 16S rRNA gene sequence similarities. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GTF13T formed a phylogenetic lineage with the family Litoricolaceae, but the genome-based phylogenomic tree showed that strain GTF13T formed a distinct phylogenetic lineage within the order Oceanospirillales. The very low 16S rRNA gene sequence similarities and distinct phylogenetic relationships, together with distinct phenotypic and chemotaxonomic properties, served to differentiate strain GTF13T from phylogenetically closely related families. Here, strain GTF13T is proposed as a novel genus and species, for which the name Aestuariirhabdus litorea gen. nov., sp. nov. is proposed, within a new family Aestuariirhabdaceae fam. nov. of the order Oceanospirillales. The type strain is GTF13T (=KACC 19788T=JCM 32043T).

Halomonas urmiana sp. nov., a moderately halophilic bacterium isolated from Urmia Lake in Iran.

In the course of screening halophilic bacteria in Urmia Lake in Iran, which is being threatened by dryness, a novel Gram-negative, moderately halophilic, heterotrophic and short rod-shaped bacteria was isolated and characterized. The bacterium was isolated from a water specimen and designated as TBZ3T. Colonies were found to be creamy yellow, with catalase- and oxidase-positive activities. The growth of strain TBZ3T was observed to be at 10-45 °C (optimum, 30 °C), at pH 6.0-9.0 (optimum, pH 7.0) and in the presence of 0.5-20 % (w/v) NaCl (optimum, 7.5 %). Strain TBZ3T contained C16 : 0, cyclo-C19 : 0 ω8c, summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c) and summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c) as major fatty acids and ubiquinone-9 as the only respiratory isoprenoid quinone. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, glycolipid, unidentified phospholipid and unidentified polar lipids were detected as the major polar lipids. Strain TBZ3T was found to be most closely related to Halomonas saccharevitans AJ275T , Halomonas denitrificans M29T and Halomonas sediminicola CPS11T with the 16S rRNA gene sequence similarities of 98.93, 98.15 and 97.60 % respectively and in phylogenetic analysis strain TBZ3T grouped with Halomonas saccharevitans AJ275T contained within a large cluster within the genus Halomonas. Based on phenotypic, chemotaxonomic and molecular properties, strain TBZ3T represents a novel species of the Halomonas genus, for which the name Halomonas urmiana sp. nov. is proposed. The type strain is TBZ3T (=DSM 22871T=LMG 25416T).