Chryseobacterium gotjawalense sp. nov. Isolated from Soil in the Volcanic Forest Gotjawal, Jeju Island.

Strain wdc7T, a rod-shaped bacterium, was isolated from soil in the Gotjawal Forest on Jeju Island, South Korea. Strain wdc7T was Gram stain-negative, facultatively anaerobic, catalase- and oxidase positive, yellow pigmented, and non-flagellated. It grew at 4-37 °C and pH 5.0-8.0 in 0-3% (w/v) NaCl. 16S rRNA gene sequencing analysis revealed that strain wdc7T belonged to the genus Chryseobacterium and was most closely related to Chryseobacterium salivictor NBC 122T, with a sequence similarity of 98.51%. Menaquinone 6 was the sole respiratory quinone, and C15:0 anteiso, C15:0 iso, and summed feature 9 were the major fatty acids. The genome length was 3.30 Mbp, with a 37% G + C content. Average amino acid identity, average nucleotide identity, and digital DNA-DNA relatedness between strain wdc7T and C. salivictor NBC 122T were 93.52%, 92.80%, and 49.7%, respectively. Digital genomic and polyphasic analyses showed that strain wdc7T likely represented a new species of the genus Chryseobacterium. We proposed the name Chryseobacterium gotjawalense sp. nov., with wdc7T (= KCTC 92440T = JCM 35602T) as the type strain.

Sulfitobacter albidus sp. nov., isolated from marine sediment of Jeju Island.

Strain JK7-1T isolated from marine sediment collected from Jeju Island of South Korea was strictly aerobic, Gram-stain-negative, catalase-positive, oxidase-positive, motile, and rod-shaped bacterium that is circular and convex with white pigment. Strain JK7-1T could grow at 10-30 °C and pH 6-9 with 1-6% (w/v) NaCl. Phylogenetic analysis based on its 16S rRNA gene sequence indicated that strain JK7-1T belonged to genus Sulfitobacter, sharing high sequence similarities with Sulfitobacter undariae W-BA2T (97.90%), Sulfitobacter donghicola KCTC 12864T (97.61%), and Sulfitobacter mediterraneus KCTC 32188T (97.47%). Strain JK7-1T possessed only ubiquinone-10 (Q-10) as a sole respiratory quinone and summed feature 8 as the major fatty acid (81.02%). A dominant polar lipid phosphatidylglycerol was identified in strain JK7-1T. Strain JK7-1T had a complete genome of 3,441,674 bp in length with a mean G + C content of 63.96%. Polyphasic and genomic analyses revealed that strain JK7-1T represented a novel species in the genus Sulfitobacter, for which a name of Sulfitobacter albidus sp. nov. was proposed. Its type strain is JK7-1T (= KCTC 72819T = NBRC 114632T = KMM 6821T).

Genomic sequence of the non-pathogen Neisseria sp. strain MA1-1 with antibiotic resistance and virulence factors isolated from a head and neck cancer patient.

Recent research has claimed virulence factors or antimicrobial resistance in commensal or non-pathogenic Neisseria spp. This study aimed to isolate and analyze commensal microorganisms related to the genus Neisseria from the oral cavity of a patient with head and neck cancer. We successfully isolated strain MA1-1 and identified its functional gene contents. Although strain MA1-1 was related to Neisseria flava based on 16S rRNA gene sequence similarity, genomic relatedness analysis revealed that strain MA1-1 was closely related to Neisseria mucosa, reported as a commensal Neisseria species. The strain MA1-1 genome harbored genes for microaerobic respiration and the complete core metabolic pathway with few transporters for nutrients. A number of genes have been associated with virulence factors and resistance to various antibiotics. In addition, the comparative genomic analysis showed that most genes identified in the strain MA1-1 were shared with other Neisseria spp. including two well-known pathogens, Neisseria gonorrhoeae and Neisseria meningitidis. This indicates that the gene content of intra-members of the genus Neisseria has been evolutionarily conserved and is stable, with no gene recombination with other microbes in the host. Finally, this study provides more fundamental interpretations for the complete gene sequence of commensal Neisseria spp. and will contribute to advancing public health knowledge.

Parasphingorhabdus halotolerans sp. nov. isolated from marine sediment in Jeju Island.

A Gram-stain-negative, catalase- and oxidase-positive, rod-shaped bacterium, designated as JK6T was isolated from a coastal marine sediment in Jeju Island. Strain JK6T was characterized by polyphasic investigation including genome features. It grew at pH 5.0-9.0 (optimum 7.5), 18-30 °C (optimum 25 °C) and 1.0-7.0% (w/v) NaCl (optimum 2.0%). Strain JK6T utilized D-mannose, D-glucose, L-fucose, propionate and acetate as carbon and energy sources. The sole quinone was ubiquinone-10, and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, and sphingolipid. Strain JK6T was closely related to Parasphingorhabdus flavimaris SW-151T (98.2%), Parasphingorhabdus marina DSM 22363T (97.6%) and Parasphingorhabdus litoris FR1093T (97.6%) based on 16S rRNA gene sequence similarity. Genome length and GC content were 3.29 Mbp and 53.0%, respectively. Digital DNA-DNA relatedness, average nucleotide identity, and average amino acid identity between strain JK6T and P. flavimaris SW-151T were 16.6%, 73.9%, and 77.6%, respectively. These results showed that the strain can be recognized as a novel bacterium named Parasphingorhabdus halotolerans. The type strain of Parasphingorhabdus halotolerans sp. nov. is JK6T (= KCTC 72818T = VTCC 910111T).

Kineobactrum salinum sp. nov., isolated from marine sediment.

Strain M2T, isolated from marine sediment collected at Jeju Island, was an aerobic, Gram-stain-negative, oxidase- and catalase-positive, motile, rod-shaped bacterium that formed circular, raised, yellow colonies. Strain M2T grew at 15-42 °C, pH 5.5-9.0 and with 1-9 % (w/v) NaCl. Phylogenetic analysis based on its 16S rRNA gene sequences indicated that strain M2T was closely related to Kineobactrum sediminis F02T (98.0 % sequence similarity). Ubiquinone-8 was determined to be the sole respiratory quinone. Summed feature 3 (C16 : 1 ω6c/C16 : 1 ω7c) and summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c) were identified as the predominant fatty acids. The DNA G+C content and digital DNA-DNA relatedness between strain M2T and K. sediminis F02T were 60.7 mol% and 19.5 %, respectively. Phosphatidylglycerol and phosphatidylethanolamine were identified as the major polar lipids. Thus, polyphasic characterization revealed that strain M2T represents a novel species in the genus Kineobactrum, for which the name Kineobactrum salinum sp. nov. is proposed. The type strain is M2T (=KCTC 72815T=VTCC 910108T).

Draconibacterium halophilum sp. nov., A Halophilic Bacterium Isolated from Marine Sediment.

A Gram-stain-negative, long-rod shaped, and facultatively anaerobic bacterium, designated as strain M1T, was isolated from the marine sediment of Jeju Island, South Korea. Strain M1T was found to be catalase- and oxidase-positive, light yellow-pigmented, non-motile, and non-flagellated, growing optimally at 30 °C, pH 7.0, and in the presence of 3% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain M1T belongs to the genus Draconibacterium and is closely related to Draconibacterium orientale FH5T (97.2%), Draconibacterium sediminis JN14CK-3 T (96.5%), "Draconibacterium filum" F2T (96.5%) and Draconibacterium mangrovi GM2-18 T (96.3% sequence similarity). The values for digital DNA-DNA hybridization ranged from 37.6 to 38.3% against D. orientale FH5T, D. sediminis KN14CK-3 T, and D. mangrovi GM2-18 T, clearly indicating that strain M1T represents a distinct species of the genus Draconibacterium. Strain M1T has a 40.0% G + C content estimated by genome sequence, menaquinone 7 as the sole respiratory quinone, C15:0 anteiso and C15:0 iso as the major fatty acids, and phosphatidylethanolamine, an unidentified phospholipid, and unidentified lipids as the polar lipids. Based on the polyphasic characteristics, it is suggested that strain M1T be assigned to the genus Draconibacterium as the type strain of a novel species, for which the name Draconibacterium halophilum sp. nov. is proposed. The type strain is M1T (= KCTC 72809 T = VTCC 910107 T).

Salinimonas marina sp. nov. Isolated from Jeju Island Marine Sediment.

A Gram-stain-negative, rod-shaped, and strictly aerobic bacterium designated strain G2-bT was isolated from the marine sediment around Jeju Island, South Korea. Strain G2-bT was found to be catalase- and oxidase-positive, white-pigmented, motile with polar flagellum, and to grow optimally at 25 °C, pH 7.0 in the presence of 4% (w/v) NaCl. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain G2-bT belongs to the genus Salinimonas and was closely related Salinimonas sediminis N102T (96.7% sequence similarity), Salinimonas iocasae KX18D6T (95.4%), Salinimonas lutimaris DPSR-4T (94.7%), and Salinimonas chungwhensis BH030046T (94.6%). Strain G2-bT possessed ubiquinone 8 as the sole respiratory quinone, summed feature 3 and summed feature 8 as the major fatty acids, and phosphatidylethanolamine and phosphatidylglycerol as the major polar lipids. The genome size and G + C content of the strain G2-bT were determined to be 3,765,169 bp, and 49.7%, respectively, as a complete circular genome. Based on the genomic analyses (e.g., average nucleotide identity and digital DNA-DNA hybridization), the strain G2-BT likely represents a new species in the genus Salinimonas, for which we propose to name this novel bacterium Salinimonas marina sp. nov., and the type strain is designated G2-BT (= KCTC 72817T = VTCC 910110T).

Expanded Diversity and Metabolic Versatility of Marine Nitrite-Oxidizing Bacteria Revealed by Cultivation- and Genomics-Based Approaches.

Nitrite-oxidizing bacteria (NOB) are ubiquitous and abundant microorganisms that play key roles in global nitrogen and carbon biogeochemical cycling. Despite recent advances in understanding NOB physiology and taxonomy, currently very few cultured NOB or representative NOB genome sequences from marine environments exist. In this study, we employed enrichment culturing and genomic approaches to shed light on the phylogeny and metabolic capacity of marine NOB. We successfully enriched two marine NOB (designated MSP and DJ) and obtained a high-quality metagenome-assembled genome (MAG) from each organism. The maximum nitrite oxidation rates of the MSP and DJ enrichment cultures were 13.8 and 30.0 µM nitrite per day, respectively, with these optimum rates occurring at 0.1 mM and 0.3 mM nitrite, respectively. Each enrichment culture exhibited a different tolerance to various nitrite and salt concentrations. Based on phylogenomic position and overall genome relatedness indices, both NOB MAGs were proposed as novel taxa within the Nitrospinota and Nitrospirota phyla. Functional predictions indicated that both NOB MAGs shared many highly conserved metabolic features with other NOB. Both NOB MAGs encoded proteins for hydrogen and organic compound metabolism and defense mechanisms for oxidative stress. Additionally, these organisms may have the genetic potential to produce cobalamin (an essential enzyme cofactor that is limiting in many environments) and, thus, may play an important role in recycling cobalamin in marine sediment. Overall, this study appreciably expands our understanding of the Nitrospinota and Nitrospirota phyla and suggests that these NOB play important biogeochemical roles in marine habitats.IMPORTANCE Nitrification is a key process in the biogeochemical and global nitrogen cycle. Nitrite-oxidizing bacteria (NOB) perform the second step of aerobic nitrification (converting nitrite to nitrate), which is critical for transferring nitrogen to other organisms for assimilation or energy. Despite their ecological importance, there are few cultured or genomic representatives from marine systems. Here, we obtained two NOB (designated MSP and DJ) enriched from marine sediments and estimated the physiological and genomic traits of these marine microbes. Both NOB enrichment cultures exhibit distinct responses to various nitrite and salt concentrations. Genomic analyses suggest that these NOB are metabolically flexible (similar to other previously described NOB) yet also have individual genomic differences that likely support distinct niche distribution. In conclusion, this study provides more insights into the ecological roles of NOB in marine environments.

Formosa sediminum sp. nov., a starch-degrading bacterium isolated from marine sediment.

A novel bacterium, designated strain PS13T, was isolated from marine sediment collected from the coast of Jeju Island. Strain PS13T was a Gram stain-negative, catalase- and oxidase-positive, aerobic, yellow-pigmented, motile by gliding, and rod-shaped bacterium. Strain PS13T grew optimally at 25 °C and pH 8.0 and in the presence of 3 % (w/v) NaCl. Results of phylogenetic analysis based on 16S rRNA gene sequences showed that strain PS13T belonged to the genus Formosa and was closely related to Formosa algae KMM3553T (98.3 % sequence similarity). The DNA-DNA relatedness (17.3-21.8 %) and average nucleotide identity (83.6-84.6 %) values clearly indicated that strain PS13T represents a distinct species of the genus Formosa. The major fatty acids were C15 : 0 iso, C16 : 1 ω6c/C16 : 1 ω7c and C15 : 1 iso G. The genomic DNA G+C content of the strain PS13T was 32.2 mol%. On the basis of polyphasic characteristics, it is suggested that strain PS13T be assigned to the genus Formosa as the type strain of a novel species, for which the name Formosa sediminum PS13T (=KCCM 43301T=CECT 9918T) sp. nov. is proposed.

Ferrovibrio terrae sp. nov., isolated from soil.

Designated strain K5T was isolated from soil on Jeju Island. The bacterium was aerobic, Gram-stain-negative, oxidase-positive, catalase-low activity, motile, short-rod shaped, opaque and formed white colonies that were circular, raised and had entire margins. Strain K5T was able to grow at 15-40 °C, pH 4-9 and at 0-2 % (w/v) NaCl concentration. Phylogenetic analysis based on its 16S rRNA gene sequences indicated that strain K5T is closely related to Ferrovibrio soli A15T (98.9 % sequence similarity), Ferrovibrio denitrificans Sp-1T (98.7 %) and Ferrovibrio xuzhouensis LM-6T (97.4 %). The sole respiratory quinone was determined to be ubiquinone-10. The dominant fatty acids of strain K5T were summed feature 8 (C18 : 1 ω7c / C18 : 1ω6c, 29.8 %), C19 : 0 cyclo ω8c (20.2 %) and C16 : 0 (24.4 %). DNA G+C content was 63.6 % and DNA-DNA relatedness between strain K5T and other three members of the genus Ferrovibrio ranged from 24 to 28 %. The major polar lipids were identified as phosphatidylglycerol, phosphatidylethanolamine and aminolipids. Moreover, polyphasic characterization revealed that strain K5T represents a novel species in the genus Ferrovibrio, for which the name Ferrovibrio terrae sp. nov. is proposed. The type strain is K5T (=KCCM 43295T=LMG 30611T).

Pusillimonas thiosulfatoxidans sp. nov., a thiosulfate oxidizer isolated from activated sludge.

A Gram-stain-negative, motile bacterium, designated strain YE3T, was isolated from activated sludge obtained from a municipal wastewater treatment plant in Daejeon Metropolitan City, Republic of Korea. The cells were oxidase- and catalase-positive, and grew under aerobic conditions at 10-40 °C (optimum, 30 °C), with 1.0-8.0 % (w/v) NaCl (1.0 %) and at pH 5.5-9.0 (pH 7.0). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YE3T was most closely related to Pusillimonasharenae KACC 14927T (98.2 % sequence similarity) and Pusillimonasginsengisoli KCTC 22046T (98.0 %). DNA-DNA relatedness values for strain YE3T and P. harenae KACC 14927T, P. ginsengisoli KCTC 22046T and P. soli KCTC 22455T were 28.7±2.27 %, 21.3±1.16 %, and 14.0±0.67 %, respectively. The genomic G+C content of the type strain YE3T was 59.3 mol%, as determined by whole-genome sequencing. The dominant fatty acids were C16 : 0 (39.2 %) and C17 : 0cyclo (37.5 %). The major polar lipids of strain YE3T were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Two aminophospholipids and four unidentified lipids were also detected. Furthermore, strain YE3T was able to oxidize thiosulfate under heterotrophic conditions. Based on the phenotypic, genotypic, chemotaxonomic and phylogenetic analyses, strain YE3T represents a novel species of the genus Pusillimonas, for which the name Pusillimonas thiosulfatoxidans sp. nov. is proposed. The type strain is YE3T (=KCTC 62737T=NBRC 113113T).

Hydrogen peroxide detoxification is a key mechanism for growth of ammonia-oxidizing archaea.

Ammonia-oxidizing archaea (AOA), that is, members of the Thaumarchaeota phylum, occur ubiquitously in the environment and are of major significance for global nitrogen cycling. However, controls on cell growth and organic carbon assimilation by AOA are poorly understood. We isolated an ammonia-oxidizing archaeon (designated strain DDS1) from seawater and used this organism to study the physiology of ammonia oxidation. These findings were confirmed using four additional Thaumarchaeota strains from both marine and terrestrial habitats. Ammonia oxidation by strain DDS1 was enhanced in coculture with other bacteria, as well as in artificial seawater media supplemented with α-keto acids (e.g., pyruvate, oxaloacetate). α-Keto acid-enhanced activity of AOA has previously been interpreted as evidence of mixotrophy. However, assays for heterotrophic growth indicated that incorporation of pyruvate into archaeal membrane lipids was negligible. Lipid carbon atoms were, instead, derived from dissolved inorganic carbon, indicating strict autotrophic growth. α-Keto acids spontaneously detoxify H2O2 via a nonenzymatic decarboxylation reaction, suggesting a role of α-keto acids as H2O2 scavengers. Indeed, agents that also scavenge H2O2, such as dimethylthiourea and catalase, replaced the α-keto acid requirement, enhancing growth of strain DDS1. In fact, in the absence of α-keto acids, strain DDS1 and other AOA isolates were shown to endogenously produce H2O2 (up to ∼4.5 µM), which was inhibitory to growth. Genomic analyses indicated catalase genes are largely absent in the AOA. Our results indicate that AOA broadly feature strict autotrophic nutrition and implicate H2O2 as an important factor determining the activity, evolution, and community ecology of AOA ecotypes.

Marinobacter salinus sp. nov., a moderately halophilic bacterium isolated from a tidal flat environment.

Two Gram-stain-negative, aerobic, motile, halophilic, rod-shaped bacteria, designated Hb8T and Hb20, were isolated from a tidal flat environment located on the South-West Korean peninsula. The isolates grew at 10-37 °C, at pH 5.0-9.0 and in NaCl concentrations of 0.5-15 % (w/v; optimum, 3.0-6.0 %). Sequence analysis of the 16S rRNA indicated that the isolates belong to the genus Marinobacter and are most closely related to Marinobacter sediminumR65T (98.3 %), followed by Marinobacter lipolyticus SM19T, Marinobacter salsuginis SD-14BT and Marinobacter similis A3d10T. The overall 16S rRNA gene sequence similarity with these species was 97.9 %, but Hb8T and Hb20 showed 100 % sequence similarity with each other. DNA-DNA relatedness values of H8T and Hb20 suggested that these isolates represent a single species, while DNA-DNA relatedness values of the two novel isolates with M. sediminum DSM 27079T and M. similis DSM 15400T were only 21.3 and 22.9 %, respectively. The major fatty acids present in strain Hb8T were identified as C16 : 0, C16 : 1ω9c, C18 : 1ω9c, C18 : 0 3-OH and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c). Ubiquinone-9 was the main respiratory quinone in both the novel strains. The polar lipids found to be present included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, four unidentified phospholipids and five unidentified lipids. The genomic DNA G+C content of Hb8T and Hb20 was 54.5 mol%. Polyphasic analysis indicated that the two isolates are representatives of a novel species of the genus Marinobacte, for which the name Marinobacter salinus sp. nov. is proposed. The type strain is Hb8T (=KCTC 52255T=JCM 31416T).

Halomonas aestuarii sp. nov., a moderately halophilic bacterium isolated from a tidal flat.

Strain Hb3T was isolated from a tidal flat in Jeollabuk-do Gunsan, Republic of Korea. Cells were Gram-stain-negative, oxidase- and catalase-positive, rod-shaped and motile. The strain grew optimally at 25-35 °C, at pH 6.0-6.5 and with 3.0-10.0 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain Hb3T belonged to the genus Halomonas. Strain Hb3T was related most closely to Halomonas ventosae Al12T (98.6 % 16S rRNA gene sequence similarity), Halomonas denitrificans M29T (98.6 %) and Halomonas saccharevitans AJ275T (98.4 %). Moreover, multilocus sequence analysis using the gyrB, rpoD and secA genes supported the phylogenetic position of strain Hb3T. The genomic G+C content of strain Hb3T was 67.9 mol%. DNA-DNA hybridization values for strain Hb3T versus H. ventosae Al12T, H. denitrificans M29T and H. saccharevitans AJ275T were 38.0, 54.5 and 47.4 %, respectively. The major quinone was ubiquinone Q-9 and the major fatty acids were C18 : 1ω7c, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), C16 : 0 and C19 : 0 cyclo ω8c. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, amino lipid, six unidentified phospholipids and an unidentified lipid comprised the polar lipid profile. On the basis of the data presented in this report, strain Hb3T represents a novel species of the genus Halomonas. The name Halomonas aestuarii sp. nov. is proposed for this novel species. The type strain is Hb3T (=KCTC 52253T=JCM 31415T).

Haloplanus salinarum sp. nov., an extremely halophilic archaeon isolated from a solar saltern.

An extremely halophilic archaeal strain SP28T was isolated from the Gomso solar saltern, Republic of Korea. Cells of the new strain SP28T were pleomorphic and Gram stain negative, and produced red-pigmented colonies. These grew in medium with 2.5-4.5 M NaCl (optimum 3.1 M) and 0.05-0.5 M MgCl2 (optimum 0.1 M), at 25-50 °C (optimum 37 °C) and at a pH of 6.5-8.5 (optimum pH 8.0). Mg2+ was required for growth. A concentration of at least 2 M NaCl was required to prevent cell lysis. Polar lipids included phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and one glycolipid chromatographically identical to sulfated mannosyl glucosyl diether. 16S rRNA and rpoB' gene sequence analyses showed that strain SP28T is closely related to Haloplanus ruber R35T (97.3 and 94.1 %, 16S rRNA and rpoB' gene sequence similarity, respectively), Haloplanus litoreus GX21T (97.0 and 92.1 %), Haloplanus salinus YGH66T (96.0 and 91.9 %), Haloplanus vescus RO5-8T (95.9 and 90.9 %), Haloplanus aerogenes TBN37T (95.6 and 90.3 %) and Haloplanus natans RE-101T (95.3 and 89.8 %). The DNA G+C content of the novel strain SP28T was 66.2 mol%, which is slightly higher than that of Hpn.litoreus GX21T (65.8 mol%) and Hpn.ruber R35T (66.0 mol%). DNA-DNA hybridization values betweenHpn.ruber R35T and strain SP28T and between Hpn.litoreus GX21T and strain SP28T were about 24.8 and 20.7 %, respectively. We conclude that strain SP28T represents a novel species of the genus Haloplanus and propose the name Haloplanus salinarum sp. nov. The type strain is SP28T (=JCM 31424T=KCCM 43210T).

Bacteroides koreensis sp. nov. and Bacteroides kribbi sp. nov., two new members of the genus Bacteroides.

Three bacterial isolates from human faeces, YS-aM39T, R2F3-3-3T and R2F3-5-1, were characterized as Gram-negative, strictly anaerobic, non-spore-forming, non-motile, and rod-shaped. Isolate YS-aM39T formed a distinct line of descent, showing greatest 16S rRNA gene sequence relatedness with R2F3-3-3T (97.5 %), R2F3-5-1 (97.5 %), Bacteroides ovatus (98.8 %) and Bacteroides xylanisolvens (97.2 %). Isolates R2F3-3-3T and R2F3-5-1 also formed a distinct line of descent, sharing greatest 16S rRNA gene sequence relatedness with B. ovatus (98.2 %) and B. xylanisolvens (97.2 %). The DNA G+C content of YS-aM39T was 44.8 mol%, that of R2F3-3-3T was 42.4 mol% and that of R2F3-5-1 was 42.6 mol%. The respiratory quinone of all three isolates was menaquinone MK-10. Polar lipid analysis identified phosphatidylethanolamine as the major lipid. The predominant fatty acids in all three isolates were anteiso-C15 : 0, iso-C15 : 0, C16 : 0 3-OH and iso-C17 : 0 3-OH. The major end products of glucose fermentation were acetic acid, lactic acid and formic acid. DNA-DNA hybridization data indicated that two isolates, YS-aM39T and R2F3-3-3T, represent a species distinct from B. ovatus and B. xylanisolvens. Finally, in this study, the two isolates represented two new species in the genus Bacteroides, for which we propose the names Bacteroides koreensis sp. nov. (type strain, YS-aM39T=KCTC 15520T=JCM 31393T) and Bacteroides kribbi sp. nov. (type strain, R2F3-3-3T=KCTC 15460T=JCM 31391T).

Detection and Diversity of the Nitrite Oxidoreductase Alpha Subunit (nxrA) Gene of Nitrospina in Marine Sediments.

Nitrite-oxidizing bacteria (NOB) are chemolithoautotrophs that catalyze the oxidation of nitrite to nitrate, which is the second step of aerobic nitrification. In marine ecosystems, Nitrospina is assumed to be a major contributor to nitrification. To date, two strains of Nitrospina have been isolated from marine environments. Despite their ecological relevance, their ecophysiology and environmental distribution are understudied owing to fastidious cultivation techniques and the lack of a sufficient functional gene marker. To estimate the abundance, diversity, and distribution of Nitrospina in various marine sediments, we used nxrA, which encodes the alpha subunit of nitrite oxidoreductase, as a functional and phylogenetic marker. We observed that Nitrospina diversity in polar sediments was significantly lower than that of non-polar samples. Moreover, nxrA-like sequences revealed an unexpected diversity of Nitrospina, with approximately 41,000 different sequences based on a 95% similarity cutoff from six marine sediments. We detected nxrA gene copy numbers of up to 3.57 × 104 per gram of marine sediment sample. The results of this study provide insight into the distribution and diversity of Nitrospina, which is fundamentally important for understanding their contribution to the nitrogen cycle in marine sediments.

Halostella salina gen. nov., sp. nov., an extremely halophilic archaeon isolated from solar salt.

A novel halophilic archaeon designated strain CBA1114T was isolated from solar salt in the Republic of Korea. Strain CBA1114T, cells of which were coccoid and Gram-stain-negative, grew in the presence of 15-30 % (w/v) NaCl (optimum, 20 %) and at 20-50 °C (optimum, 40 °C) and pH 7.0-9.0 (optimum, pH 8.0). Strain CBA1114T required Mg2+ for growth. Strain CBA1114T had three 16S rRNA genes, rrnA, rrnB and rrnC; levels of similarity between the sequences were 99.7-99.9 %. The 16S rRNA gene sequence of strain CBA1114T showed 91.7 % similarity to that of Haloterrigena thermotolerans PR5T. In multilocus sequence analysis (MLSA), five housekeeping genes, atpB, EF-2, radA, rpoB' and secY, were found to be closely related to those of the members of the genera Halorientalis(89.7 % similarity of the atpB gene sequence), Halomicroarcula(91.9 %, EF-2), Haloterrigena(85.4 %, radA), Natronoarchaeum(89.2 %, rpoB') and Natrinema(75.7 %, secY). A phylogenetic tree generated from the results of MLSA of the five housekeeping genes showed that strain CBA1114T was closely related to species of the genus Halorientalis in the family Halobacteriaceae. The major polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and unidentified lipids. The G+C content of the genomic DNA of strain CBA1114T was 68.1 mol%. According to the results of phylogenetic, phenotypic and chemotaxonomic analyses, we designate strain CBA1114T (=JCM 30111T=KCTC 4206T) as the type strain of Halostella salina gen. nov., sp. nov., a novel species of a new genus within the family Halobacteriaceae.

Halolamina sediminis sp. nov., an extremely halophilic archaeon isolated from solar salt.

An extremely halophilic archaeal strain, halo-7T, was isolated from brine sediment of the Gomso solar saltern, Republic of Korea. Cells of strain halo-7T were pleomorphic, stained Gram-negative, lysed in distilled water and formed red-pigmented colonies. Strain halo-7T grew in the range of 25-45°C (optimum 37-40°C), pH 6.5-9.5 (optimum pH 7.0-8.0), 15-30% (w/v) NaCl (optimum, 20-25%), and 0.05-0.5 M MgCl2 (optimum 0.1-0.3 M). The minimal NaCl concentration to prevent cell lysis of strain halo-7T was 10% (w/v). The major polar lipids of the isolate were phosphatidylglycerol, phosphatidylglycerolphosphate methyl ester, an unidentified phospholipid, an unidentified lipid, and two unidentified glycolipids. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain halo-7T is closely related to the members of the genus Halolamina, Halolamina salina WSY15-H3T (98.7% 16S rRNA gene sequence similarity), Halolamina pelagica TBN21T (98.2%) and Halolamina rubra CBA1107T (97.4%). The genomic DNA G+C content determined for strain halo-7T (68.0 mol%) was slightly higher than those of H. salina JCM 18549T and H. rubra CBA1107T. DNA-DNA hybridization values between strain halo-7T and reference strains were < 25%. Based on phenotypic, chemotaxonomic and phylogenetic properties, we describe a novel species of the genus Halolamina, represented by strain halo-7T, for which we propose the name Halolamina sediminis sp. nov. The type strain is halo-7T ( = JCM 30187T = CECT 8739T).

Rhodanobacter aciditrophus sp. nov., an acidophilic bacterium isolated from mine wastewater.

A novel strain (designated sjH1T), characterized as aerobic, Gram-stain-negative, oxidase-positive, catalase-negative, motile and rod-shaped, was isolated from mine wastewater. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain sjH1T belonged to the genus Rhodanobacter. Strain sjH1T was closely related to Rhodanobacter thiooxydans LCS2T (98.0% 16S rRNA gene sequence similarity), Rhodanobacter denitrificans 2APBS1T (97.7%), Rhodanobacter soli DCY45T (97.2%) and Rhodanobacter caeni MJ01T (97.0%). The DNA G+C content of strain sjH1T was 69.2 mol%. DNA-DNA relatedness ( < 60%) indicated that strain sjH1T represents a distinct species that is separate from R. thiooxydans, R. denitrificans, R. soli and R. caeni. The major ubiquinone was Q-8, and major fatty acids were summed feature 9 (iso-C17 : 1ω9c and/or C16 : 0 10-methyl), iso-C15 : 0, iso-C17 : 0, iso-C16 : 0 and anteiso-C15 : 0. Based on data from this polyphasic study, it is proposed that sjH1T ( = KCTC 42660T = JCM 30774T) is the type strain of a novel species, Rhodanobacter aciditrophus sp. nov.