- Volume 66, Issue 3, 2016

A Gram-stain-positive, aerobic, coccus-shaped, non-spore-forming actinobacterium, designated strain 2Q3S-4-2T, was isolated from the surface-sterilized bark of Kandelia candel, collected from Cotai Ecological Zones in Macao, PR China. It was tested using a polyphasic approach to determine its taxonomic position. Strain 2Q3S-4-2T grew optimally without NaCl at 28–30 °C and at pH 7.0. Substrate mycelia and aerial mycelia were not formed and no diffusible pigments were observed on the media tested. Phylogenetic analysis, based on 16S rRNA gene sequences, suggested that strain 2Q3S-4-2T belonged to the genus Nakamurella, sharing highest 16S rRNA gene sequence similarity with Nakamurella flavida DS-52T (96.76 %). The DNA G+C content of strain 2Q3S-4-2T was 67.8 mol%. The cell-wall peptidoglycan contained meso-diaminopimelic acid and MK-8(H4) was the predominant menaquinone. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, unidentified aminophospholipids and phosphatidylinositol. The major fatty acids were iso-C16 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and C16 : 0. On the basis of the phylogenetic, phenotypic and chemotaxonomic analysis, strain 2Q3S-4-2T represents a novel species of the genus Nakamurella, for which the name Nakamurella endophytica sp. nov. is proposed. The type strain is 2Q3S-4-2T ( = DSM 100722T = CGMCC 4.7308T).

A Gram-stain-variable, rod-shaped, motile bacterial strain, designated S1-20T, was isolated from marine sediment of the Yellow Sea in China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain S1-20T was related to the genus Virgibacillus and had highest 16S rRNA gene sequence similarities with the type strains of Virgibacillus necropolis (98.1 %), Virgibacillus arcticus (97.7 %) and Virgibacillus carmonensis (97.3 %). The predominant cellular fatty acids of strain S1-20T were anteiso-C15 : 0 and anteiso-C17 : 0. The main menaquinone was MK-7. The genomic DNA G+C content of strain S1-20T was 38.3 mol%. The polar lipids profile contained diphosphatidylglycerol, phosphatidylglycerol, five unidentified phospholipids, one unknown aminophospholipid and an aminolipid. Combined data from phenotypic, phylogenetic and DNA–DNA relatedness studies demonstrated that strain S1-20T is a representative of a novel species of the genus Virgibacillus, for which the name Virgibacillus flavescens sp. nov. is proposed. The type strain is S1-20T ( = LMG 28381T = DSM 29015T).

A Gram-reaction-positive, spore-forming, rod-shaped bacterium, designated NR1-3-2T, was isolated from fish sauce collected from a factory in Chonburi province, Thailand. Strain NR1-3-2T grew at pH 5–10 (optimum 7.5), at 21–48 °C (optimum 37 °C) and with 0–15 % (w/v) NaCl (optimum 1–3 %). The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. Menaquinone with seven isoprene units (MK-7) was the major isoprenoid quinone. The strain contained anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0 as major cellular fatty acids. Diphosphatidylglycerol, phosphatidyl glycerol and one unknown glycolipid were detected as major polar lipids. On the basis of 16S rRNA gene sequence analysis, strain NR1-3-2T belonged to the genus Bacillus and was closely related to Bacillus iranensis DSM 23995T (97.4 % similarity). Strain NR1-3-2T exhibited low DNA–DNA relatedness (31.2–39.8 %) with B. iranensis DSM 23995T. The DNA G+C content was 44.2 mol%. On the basis of phenotypic characteristics, DNA–DNA relatedness and phylogenetic analyses, the strain is considered to represent a novel species of the genus Bacillus and the name Bacillus piscicola sp. nov. is proposed. The type strain is NR1-3-2T ( = JCM 19598T = LMG 28281T = PCU 340T = TISTR 2295T).

A Gram-stain-positive, facultatively anaerobic, rod-shaped and endospore-forming bacterium, strain BK114-2T isolated from tree bark in Thailand was characterized taxonomically using a polyphasic approach. Analysis based on comparison of 16S rRNA gene sequences indicated that strain BK114-2T was affiliated to the genus Paenibacillus, and was closely related to Paenibacillus timonensis 2301032T (96.7 % 16S rRNA gene sequence similarity), Paenibacillus phoenicis 3PO2SAT (96.6 %) and Paenibacillus barengoltzii SAFN-016T (96.4 %). Strain BK114-2T contained meso-diaminopimelic acid in its cell-wall peptidoglycan. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unknown phospholipids, five unknown aminophospholipids and six unknown lipids. The only menaquinone detected was MK-7 and the dominant cellular fatty acids were C16 : 0 (22.9 %), anteiso-C15 : 0 (22.6 %), iso-C16 : 0 (19.4 %) and anteiso-C17 : 0 (14.7 %). The DNA G+C content was 52.0 mol%. Based on these results, strain BK114-2T repreesents a novel species of the genus Paenibacillus, for which the name Paenibacillus cathormii sp. nov. is proposed. The type strain is BK114-2T ( = KCTC 33251T = TISTR 2282T).

Two strains, designated DW5-4T and NH7I_1T, were isolated from aquaculture water of a shrimp farm of Zhangzhou city in China and from surface sediment of the South China Sea, respectively. Cells of the strains were Gram-stain-positive, oxidase- and catalase-positive, rod-shaped and motile. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the two strains belonged to the Bacillus pumilus clade. 16S rRNA gene sequence similarity between strains DW5-4T and NH7I_1T was 99.8 %, and between each of the two strains and four type strains, B. pumilus ATCC 7061T, Bacillus safensis FO-36bT, Bacillus altitudinis 41KF2bT and Bacillus xiamenensis HYC-10T, were above 99.4 %. The digital DNA–DNA hybridization value between the two novel strains was 41.6 %, and between each of the two strains and the four above-mentioned type strains were below 53.0 %. In both strains, the principal fatty acids were iso-C15 : 0 and anteiso-C15 : 0, and the isoprenoid quinone was identified as MK-7 (100 %). The DNA G+C contents of strains DW5-4T and NH7I_1T were 41.4 and 41.3 mol%, respectively. Based on phenotypic and genotypic characteristics, two novel species of the genus Bacillus are proposed: Bacillus zhangzhouensis sp. nov., with DW5-4T ( = MCCC 1A08372T = LMG 27144T = KCTC 33531T) as the type strain, and Bacillus australimaris sp. nov., with NH7I_1T ( = MCCC 1A05787T = LMG 27697T = KCTC 33532T) as the type strain.

Bacillus velezensis was previously reported to be a later heterotypic synonym of Bacillus amyloliquefaciens, based primarily on DNA–DNA relatedness values. We have sequenced a draft genome of B. velezensis NRRL B-41580T. Comparative genomics and DNA–DNA relatedness calculations show that it is not a synonym of B. amyloliquefaciens. It was instead synonymous with Bacillus methylotrophicus. ‘Bacillus oryzicola’ is a recently described species that was isolated as an endophyte of rice (Oryza sativa). The strain was demonstrated to have plant-pathogen antagonist activity in greenhouse assays, and the 16S rRNA gene was reported to have 99.7 % sequence similarity with Bacillus siamensis and B. methylotrophicus, which are both known for their plant pathogen antagonism. To better understand the phylogenetics of these closely related strains, we sequenced the genome of ‘B. oryzicola’ KACC 18228. Comparative genomic analysis showed only minor differences between this strain and the genomes of B. velezensis NRRL B-41580T, B. methylotrophicus KACC 13015T and Bacillus amyloliquefaciens subsp. plantarum FZB42T. The pairwise in silico DNA–DNA hybridization values calculated in comparisons between the strains were all greater than 84 %, which is well above the standard species threshold of 70 %. The results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the strains share phenotype and genotype coherence. Therefore, we propose that B. methylotrophicus KACC 13015T, B. amyloliquefaciens subsp. plantarum FZB42T, and ‘B. oryzicola’ KACC 18228 should be reclassified as later heterotypic synonyms of B. velezensis NRRL B-41580T, since the valid publication date of B. velezensis precedes the other three strains.

Twenty-three rod-shaped, endospore-forming, Gram-stain-positive, obligately anaerobic bacteria were isolated from different marine sediment samples of Gujarat. All 23 strains shared 16S rRNA gene sequence similarity of ∼100 %. Strain JC272T was designated the type strain and shared highest sequence similarity with Clostridium bifermentans ATCC 638T (99.8 %), Clostridium ghonii JCM 1400T (98.0 %), Clostridium sordellii ATCC 9714T (97.9 %) and other members of the genus Clostridium ( < 96.4 %). C16 : 0, C18 : 0, C17 : 0, C16 : 1ω9c and iso-C16 : 0 were the major (>5 %) fatty acids. Strain JC272T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and two unidentified amino lipids. Genome-based analysis of average nucleotide identity (ANI) and in silico DNA–DNA hybridization (DDH) of strain JC272T with C. bifermentans ATCC 638T yielded values of 94.35 and 58.5 ± 2.8 %, respectively. The DNA G+C content of strain JC272T was 28.3 mol%. Strain JC272T together with C. bifermentans were found to fall outside Clostridium rRNA cluster I considered as Clostridium senso stricto. Based on ANI value, in-silico DDH, and distinct morphological and physiological differences from the previously described taxa, we suggest that strain JC272T represents a novel species of a new genus in the family Clostridiaceae, for which the name Paraclostridium benzoelyticum gen. nov., sp. nov. is proposed. The type strain is JC272T ( = KCTC 15476T = LMG 28745T). It is also proposed to transfer C. bifermentans to this new genus, as Paraclostridium bifermentans comb. nov. (type strain ATCC 638T = DSM 14991T = JCM 1386T). The genus Paeniclostridium gen. nov. is proposed to accommodate C. sordellii and C. ghonii as Paeniclostridium sordellii comb. nov. (type strain ATCC 9714T = LMG 15708T = JCM 3814T) and Paeniclostridium ghonii comb. nov. (type strain ATCC 25757T = DSM 15049T = JCM 1400T).

A novel, anaerobic bacterium, strain MO-SEDIT, was isolated from a methanogenic microbial community, which was originally obtained from marine subsurface sediments collected from off the Shimokita Peninsula of Japan. Cells were Gram-stain-negative, non-motile, non-spore-forming rods, 0.4–1.4 μm long by 0.4–0.6 μm wide. The cells also formed long filaments of up to about 11 μm. The strain grew on amino acids (i.e. valine, leucine, isoleucine, methionine, glycine, phenylalanine, tryptophan, lysine and arginine), pyruvate and melezitose in the presence of yeast extract. Growth was observed at 4–37 °C (optimally at 30 °C), at pH 6.0 and 8.5 (optimally at 7.0–7.5) and in 0–60 g l− 1 NaCl (optimally 20 g NaCl l− 1). The G+C content of the DNA was 32.0 mol%. The polar lipids of strain MO-SEDIT were phosphatidylglycerol, phosphatidyl lipids and unknown lipids. The major cellular fatty acids (>10 % of the total) were C14 : 0, C16 : 1ω9 and C16 : 0 dimethyl aldehyde. Comparative sequence analysis of the 16S rRNA gene showed that strain MO-SEDIT was affiliated with the genus Sedimentibacter within the phylum Firmicutes. It was related most closely to the type strain of Sedimentibacter saalensis (94 % sequence similarity). Based on the phenotypic and genetic characteristics, strain MO-SEDIT is considered to represent a novel species of the genus Sedimentibacter, for which the name Sedimentibacter acidaminivorans sp. nov. is proposed. The type strain is MO-SEDIT ( = JCM 17293T = DSM 24004T).

A Gram-stain-positive, catalase-positive, facultatively anaerobic, spore-forming, rod-shaped bacterium, strain NK26-11T, was isolated from soil in Thailand. This strain produced d-lactic acid from glucose homofermentatively, and grew at 20–45 °C and pH 5–8.5. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The major respiratory quinone was menaquinone 7 (MK-7), the DNA G+C content was 42.6 mol%, and the major cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0. On the basis of 16S rRNA gene sequences analysis, strain NK26-11T was closely related to Bacillus solimangrovi JCM 18994T (93.89 % 16S rRNA gene sequence similarity), Pullulanibacillus naganoensis LMG 12887T (93.32 %), Sporolactobacillus inulinus NRIC 1133T (92.99 %), Tuberibacillus calidus JCM 13397T (92.98 %) and Thalassobacillus devorans DSM 16966T ( < 90.93 %). Strain NK26-11T could be clearly distinguished from the closely related genera based on phenotypic characteristics and DNA G+C content, and thus represents a novel species of a new genus between the Bacillus and Sporolactobacillus cluster, for which the name Terrilactibacillus laevilacticus gen. nov., sp. nov. is proposed. The type strain of the type species is NK26-11T ( = LMG 27803T = TISTR 2241T = PCU 335T).

A moderately halophilic bacterium was isolated from a brine sample of a hypersaline lake, Aran-Bidgol, in Iran. The strain, designated J8BT, was Gram-stain-positive, endospore-forming, rod-shaped, strictly aerobic, motile and produced cream colonies. Strain J8BT grew in NaCl at between 3.0–15.0 % (w/v) (optimally at 7.5 % NaCl, w/v), between pH 6.5–9.0 (optimally at pH 8.0) and between 20–45 °C (optimally at 35 °C). Phylogenetic analysis, based on 16S rRNA gene sequences, revealed that strain J8BT is a member of the genus Oceanobacillus and most closely related to Oceanobacillus profundus CL-MP28T, Oceanobacillus polygoni SA9T and Oceanobacillus oncorhynchi R-2T (96.9 %, 96.3 % and 96.2 % similarities, respectively). The level of DNA–DNA relatedness between the novel isolate and O. profundus IBRC-M 10567T was 10 %. The major cellular fatty acids of the isolate were anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0. The polar lipid pattern of strain J8BT consisted of phosphatidylglycerol, diphosphatidylglycerol, five phospholipids, two aminolipids and two glycoaminolipids. It contained MK-7 as the predominant menaquinone and meso-diaminopimelic acid in the cell-wall peptidoglycan. The G+C content of the genomic DNA of this strain was 39.2 mol%. Phenotypic characteristics, phylogenetic analysis and DNA–DNA relatedness data suggest that this strain represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus halophilus sp. nov. is proposed. The type strain is strain J8BT ( = IBRC-M 10444T = DSM 23996T).

A Gram-stain positive, endospore-forming, strictly anaerobic bacterium, designated strain Gal1T, was isolated from shea cake, a waste material from the production of shea butter, originating from Saraya, Senegal. The cells were rod-shaped, slightly curved, and motile with peritrichous flagella. The strain was oxidase-negative and catalase-negative. Growth was observed at temperatures ranging from 15 to 45 °C (optimum 30 °C) and at pH 6.5–9.3 (optimum pH 7.8). The salinity range for growth was 0–3.5 % NaCl (optimum 1 %). Yeast extract was required for growth. Strain Gal1T fermented various carbohydrates such as mannose, mannitol, arabinose, cellobiose, fructose, glucose, maltose, sucrose, trehalose and lactose and the major end-products were ethanol and acetate. The only major cellular fatty acid was C16 : 0 (19.6 %). The DNA base G+C content of strain Gal1T was 33.8 mol%. Analysis of the 16S rRNA gene sequence of the isolate indicated that this strain was related to Mobilitalea sibirica DSM 26468T with 94.27 % similarity, Clostridium populeti ATTC 35295T with 93.94 % similarity, and Clostridium aminovalericum DSM 1283T and Anaerosporobacter mobilis DSM 15930T with 93.63 % similarity. On the basis of phenotypic characteristics, phylogenetic analysis and the results of biochemical and physiological tests, strain Gal1T was clearly distinguished from closely related genera, and strain Gal1T can be assigned to a novel species of a new genus for which the name Mobilisporobacter senegalensis gen. nov., sp. nov. is proposed. The type strain is Gal1T ( = DSM 26537T = JCM 18753T).

Strain BD3526T, isolated from raw yak milk collected in Tibet, China, was studied to determine its taxonomic status. The strain was Gram-reaction positive, motile, catalase-positive, aerobic or facultatively anaerobic. The DNA G+C content of the strain was 47.5 mol% and its peptidoglycan contained meso-diaminopimelic acid. The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and C16 : 0. Based on 16S rRNA gene sequence analysis, similarities among strain BD3526T and its most closely related species, Paenibacillus shenyangensis A9T, ‘Paenibacillus dauci’ H9, ‘Paenibacillus wulumuqiensis’ Y24 and Paenibacillus hunanensis DSM 22170T were 99.0, 98.5, 97.3 and 96.7 %, respectively. Levels of DNA–DNA relatedness among strain BD3526T and P. hunanensis DSM 22170T and Paenibacillus polymyxa ATCC 842T ( = DSM 36T), the type species of the genus, were 41.2 and 45.6 %, respectively. In silico genome-to-genome comparison showed that the DNA–DNA hybridization values among strain BD3526T and P. shenyangensis A9T, ‘P. dauci’ H9 and ‘P. wulumuqiensis’ Y24 were lower than 70 %. Based on its molecular and physiological properties, strain BD3526T ( = DSM 28815T = CGMCC 8333T) is identified as the type strain of a novel species within the genus Paenibacillus, for which the name Paenibacillus bovis is proposed.

During investigation of the biological contamination of the 1300-year-old mural paintings and plaster walls inside the stone chambers of the Takamatsuzuka and Kitora Tumuli (TT and KT) in Asuka-mura, Nara Prefecture, Japan, the identity of 17 bacterial isolates from blackish mouldy spots and viscous gels (biofilms) collected from both tumuli (16 isolates from TT and one from KT) during our 2005–2007 microbiological survey was systematically elucidated. One cluster of the major bacterial isolates was assigned to the genus Stenotrophomonas (class Gammaproteobacteria) by phylogenetic analysis of the 16S rRNA gene sequences. These isolates were divided into two groups A and B. Group A comprised 15 TT isolates that took a phylogenetic position near Stenotrophomonas chelatiphaga LPM-5T. Based on our analysis of the phenotypic (cultural, morphological, physiological and chemotaxonomic) characteristics and genotypic/molecular characteristics (DNA base composition, DNA–DNA relatedness, and 16S rRNA and gyrB gene sequences), the novel species name Stenotrophomonas tumulicola sp. nov. is proposed for the group A isolates with the type strain T5916-2-1bT ( = JCM 30961T = NCIMB 15009T). Group B, which contained only one TT and one KT isolate, was closely related to [Pseudomonas] geniculata, [P.] hibiscicola, [P.] beteli, Stenotrophomonas maltophilia and Stenotrophomonas pavanii. The two isolates were genotypically and phenotypically assignable to S. maltophilia.

Strain NHI-13T, a Gram-stain-negative, aerobic and short rod-shaped bacterium, was isolated from forest soil at Kyonggi University in Suwon, South Korea. It grew optimally in R2A medium, at 20–30 °C, in the presence of 0–4 % NaCl. Colonies resulting from incubation of the strain on agar plates for 2 days were circular, raised, translucent, viscous and whitish-yellow, with entire margins. This strain exhibited high catalase activity but was negative for oxidase. 16S rRNA gene sequence analysis showed that strain NHI-13T formed a coherent cluster with members of the genus Brevundimonas. Its similarities were 98.0 % with Brevundimonas aurantiaca DSM 4731T, 97.9 % with Brevundimonas vesicularis LMG 2350T, 97.6 % with Brevundimonas intermedia ATCC 15262T, 97.5 % with Brevundimonas nasdae GTC 1043T, 97.1 % with ‘Brevundimonas olei’ MJ15, 97.1 % with Brevundimonas mediterranea V4.BO.10T and 97.0 % with Brevundimonas poindexterae FWC40T. The major cellular fatty acids were summed feature 8 (C18 : 1ω7c/C18 : 1ω6c), C16 : 0 and 11-methyl C18 : 1ω7c. The DNA G+C content was 63 mol%. The predominant quinone was ubiquinone Q-10. The polar lipid profile contained 1,2-di-O-acyl-3-O-α-d-glycopyranuronosyl glycerol, 1,2-di-O-acyl-3-O-α-d-glycopyranosyl glycerol, 1,2-di-O-acyl-3-O-[d-glycopyranosyl (1 → 4)-α-d-glucopyranuronosyl] glycerols, phosphatidylglycerol, 1,2-diacyl-3- O-(6′-phosphatidyl-α-d-glucopyranosyl) glycerol and other unknown lipids. The DNA relatedness of strain NHI-13T with its reference strains was in the range of 43–56 %. On the basis of its phenotypic, genotypic, chemotaxonomic and phylogenetic distinctiveness, strain NHI-13T is suggested to be a representative of a novel species, belonging to the genus Brevundimonas. Therefore, the name Brevundimonas albigilva. sp. nov. is proposed, with the type strain being NHI-13T ( = KEME 9005-016T = KACC 18249T = JCM 30385T).

A biofilm-forming, Gram-stain-negative, aerobic, catalase-positive but oxidase-negative strain, designated CT6T, was isolated from the microbial mats (∼45 °C) of a hot water spring, located within the Himalayan ranges at Manikaran, Himachal Pradesh, India. Strain CT6T formed white, smooth colonies with irregular margins. Transmission electron microscopy revealed coccoid, non-flagellated cells with wavy boundaries. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain CT6T belongs to the genus Lampropedia with a sequence similarity value of 95.4 % to the sole member of this genus, Lampropedia hyalina ATCC 11041T. Strain CT6T was found to have phosphatidylethanolamine and phosphatidylglycerol as the major polar lipids. The major cellular fatty acids were C16 : 0, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C14 : 0, C19 : 0ω8c cyclo and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The major respiratory quinone was ubiquinone-8. The major polyamines were putrescine, spermidine and the betaproteobacterial-specific 2-hydroxyputrescine. The DNA G+C content was 63.5 mol%. Based on the genotypic, phenotypic, physiological and biochemical data, strain CT6T is considered to represent a novel species of the genus Lampropedia, for which the name Lampropedia cohaerens sp. nov. is proposed ( = DSM 100029T = KCTC 42939T = MCC 2711T).

Analysis of the microbiota of raw cow's milk and semi-finished milk products yielded seven isolates assigned to the genus Pseudomonas that formed two individual groups in a phylogenetic analysis based on partial rpoD and 16S rRNA gene sequences. The two groups could be differentiated from each other and also from their closest relatives as well as from the type species Pseudomonas aeruginosa by phenotypic and chemotaxonomic characterization and average nucleotide identity (ANIb) values calculated from draft genome assemblies. ANIb values within the groups were higher than 97.3 %, whereas similarity values to the closest relatives were 85 % or less. The major cellular lipids of strains WS4917T and WS4993T were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol; the major quinone was Q-9 in both strains, with small amounts of Q-8 in strain WS4917T. The DNA G+C contents of strains WS4917T and WS4993T were 58.08 and 57.30 mol%, respectively. Based on these data, strains WS4917T, WS4995 ( = DSM 29141 = LMG 28434), WS4999, WS5001 and WS5002 should be considered as representatives of a novel species of the genus Pseudomonas, for which the name Pseudomonas helleri sp. nov. is proposed. The type strain of Pseudomonas helleri is strain WS4917T ( = DSM 29165T = LMG 28433T). Strains WS4993T and WS4994 ( = DSM 29140 = LMG 28438) should be recognized as representing a second novel species of the genus Pseudomonas, for which the name Pseudomonas weihenstephanensis sp. nov. is proposed. The type strain of Pseudomonas weihenstephanensis is strain WS4993T ( = DSM 29166T = LMG 28437T).

The taxonomic status of the bacterium Wolbachia persica is described, and based on the evidence presented, transfer of this species to the genus Francisella as Francisella persica comb. nov. is proposed. This reclassification is supported by data generated from genomic comparisons of W. persica ATCC VR-331T ( = FSC845T = DSM 101678T) to other near neighbours, including Francisella tularensis subsp. novicida. The full-length 16S rRNA gene sequence of strain ATCC VR-331T had 98.5 % nucleotide identity to the cognate gene in F. tularensis, with the highest similarity to subspecies novicida. Phylogenetic trees of full-length 16S rRNA gene, gyrA and recA sequences from species of the genera Wolbachia (class Alphaproteobacteria) and Francisella (class Gammaproteobacteria) indicated that W. persica ATCC VR-331T was most closely related to members of the genus Francisella and not Wolbachia. Local collinear blocks within the chromosome of strain ATCC VR-331T had considerable similarity with F. tularensis subsp. novicida, but not with any Wolbachia strain. The genomes of strain ATCC VR-331T and F. tularensis subsp. novicida Utah 112T ( = ATCC 15482T) contained an average nucleotide identity mean of 88.72 % and median of 89.18 %. Importantly, the genome of strain ATCC VR-331T contained one Francisella Pathogenicity Island, similar to F. tularensis subsp. novicida, as well as the Francisella-specific gene fopA1 and F. tularensis-specific genes fopA2 and lpnA (also referred to as tul4). In contrast to the obligate intracellular genus Wolbachia, strain ATCC VR-331T and facultative intracellular Francisella can replicate in specialized cell-free media. Collectively, these results demonstrate that Wolbachia persica should be reclassified in the genus Francisella as Francisella persica comb. nov. The type strain of Francisella persica comb. nov. is ATCC VR-331T ( = FSC845T = DSM 101678T). An emended description of the family Francisellaceae is also provided.

A novel psychrotolerant bacterium, designed strain M6-79T, was isolated from an arctic glacial foreland soil sample collected from Ny-Ålesund in the Svalbard Archipelago, Norway. Cells of strain M6-79T were Gram-stain-negative, rod-shaped and produced a red-pigment. Strain M6-79T was strictly aerobic, non-motile, non-endospore-forming, oxidase-negative and catalase-positive. Based on 16S rRNA gene sequence analysis, strain M6-79T was phylogenetically related to Roseomonas aquatica TR53T (95.2 % 16S rRNA gene sequence similarity), Roseomonas lacus TH-G33T (94.3 %), ‘Roseomonas sediminicola’ FW-3 (94.3 %), Roseomonas terrae DS-48T (94.1 %) and Roseomonas soli 5N26T (94.1 %). The unique isoprenoid quinone detected in strain M6-79T was Q-9. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, one unknown aminolipid and one unknown lipid. Strain M6-79T possessed C18 : 1ω7c, C16 : 0 and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) as the predominant fatty acids, and the DNA G+C content was 71.2 mol%. Combined data from phylogenetic, phenotypic and chemotaxonomic studies revealed that strain M6-79T represents a novel species of the genus Roseomonas, for which the name Roseomonas arctica sp. nov. is proposed. The type strain is strain M6-79T ( = CCTCC AB 2013101T = LMG 28251T).

A Gram-stain-negative, rod-shaped, motile bacterium, PC004T, was isolated from root nodules of the Thai medicinal plant Pueraria candollei var. candollei. 16S rRNA gene sequence analysis indicated that the strain is phylogenetically related to species in the genus Rhizobium, showing highest similarity (96.6 %) with Rhizobium mesosinicum HAMBI 3194T. The phylogenetic tree reconstructed based on 16S rRNA gene sequences showed that strain PC004T forms a cluster with Rhizobium petrolearium KCTC 23288T. Based on atpD, gyrB and recA gene sequences, strain PC004T also showed low similarity ( < 90 %) to reference strains. These phylogenetic data indicate that PC004T may represent a novel species. Strain PC004T also exhibited low DNA–DNA relatedness with R. mesosinicum HAMBI 3194T (8.2 %) and R. petrolearium KCTC 23288T (26.3 %). The DNA G+C content of strain PC004T was 64 mol%, which is within the range reported for the genus Rhizobium. The major fatty acid of PC004T was C18 : 1ω7c with minor amounts of C16 : 0, C16 : 0 3-OH, C18 : 0 3-OH, C18 : 1 2-OH, C19 : 0 cyclo ω8c and summed feature 2. The strain was able to grow at pH 12 and with up to 2 % (w/v) NaCl. Strain PC004T did not nodulate five tested legumes and the nifH and nodC genes were not detected by PCR. Based on the physiological, chemotaxonomic and phenotypic data from this study, strain PC004T represents a novel species of the genus Rhizobium, for which the name Rhizobium puerariae sp. nov. is proposed; the type strain is PC004T ( = BCC 73740T = NBRC 110722T).