A facultatively anaerobic, alkaliphilic, Gram-stain-positive, rod-shaped bacterium, designated strain 12/1T, isolated from alkaline wastewater drained sludge of a beverage industry facility located near New Delhi, India, was subjected to a polyphasic taxonomic study. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain 12/1T belonged to the genus Exiguobacterium and was most closely related to Exiguobacterium aurantiacum DSM 6208T (99.46 %), E. aquaticum IMTB-3094T (99.18 %), E. mexicanum 8NT (99.06 %), E. profundum 10CT (98.17 %), E. aestuarii TF-16T (98.1 %) and E. marinum TF-80T (98.03 %). The DNA G+C content of strain 12/1T was 55.6 mol%, major respiratory isoprenoid quinone was MK-7, major polar lipids were phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine and the cell-wall peptidoglycan was of the A3α l-Lys–Gly type, characteristics consistent with its affiliation to the genus Exiguobacterium . Strain 12/1T showed levels of DNA–DNA hybridization of less than 70 % with the closely related species of the genus Exiguobacterium . Overall, the phenotypic, chemotaxonomic and phylogenetic data presented in this study suggest that strain 12/1T represents a novel species of the genus Exiguobacterium , for which the name Exiguobacterium alkaliphilum sp. nov. is proposed. The type strain is 12/1T ( = CCM 8459T = DSM 21148T).
A bacterial strain, designated PECAE04T, was isolated from root nodules of Cicer arietinum in Spain. Phylogenetic analysis based on 16S rRNA gene sequence placed the isolate into the genus Paenibacillus with its closest relative being Paenibacillus castaneae Ch-32T with 98.4 % 16S rRNA gene sequence similarity followed by Paenibacillus glycanilyticus DS-1T, Paenibacillus prosopidis PW21T, Paenibacillus xinjiangensis B538T and Paenibacillus catalpae D75T with similarities ranging from 97.9 to 96.8 %. DNA–DNA hybridization measurements showed values lower than 20 % between the strain PECAE04T and any of these species. The isolate was a Gram-stain-positive, motile, sporulating rod. Catalase and oxidase activities were positive. Aesculin was hydrolysed but casein and gelatin were not. Acetoin production, H2S production, nitrate reduction and urease and caseinase production were negative. Growth was supported by many carbohydrates and organic acids as carbon sources. MK-7 was the predominant menaquinone and anteiso-C15 : 0, iso-C16 : 0 and C16 : 0 were the major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, a glycolipid, three phospholipids and an unidentified lipid. Meso-diaminopimelic acid was not detected in the peptidoglycan. The DNA G+C content was 52.9 mol%. Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain PECAE04T should be considered to be a representative of a novel species of the genus Paenibacillus , for which the name Paenibacillus endophyticus sp. nov. is proposed. The type strain is PECAE04T ( = LMG 27297T = CECT 8234T).
An obligately alkaliphilic, anaerobic, thermo- and halotolerant, spore-forming bacterium was isolated from sediments of soda lake Magadi (Kenya) and designated strain Z-1001T. Cells of strain Z-1001T were straight, Gram-positive rods, slowly motile. Strain Z-1001T was found to be an obligate anaerobe. It grew within a pH range from 7.5 to 10.7 with an optimum at 9.25–9.5 (at 40 °C), a temperature range from 20 to 57 °C with an optimum at 45–50 °C, and a NaCl concentration range from 0 to 1.55 M with an optimum at 1.2–1.4 M. Peptides, such as meat and yeast extracts, peptone and tryptone, were fermented by Z-1001T. Carbohydrates did not support growth. With yeast extract as an electron donor, strain Z-1001T reduced , , , Fe(III) citrate and anthraquinone-2,6-disulfonate (AQDS) as electron acceptors. The isolate was able to grow oligotrophically with a very small amount of yeast extract: 0.03 g l−1. The main fatty acids were C16 : 0, C16 : 1ω7c , C18 : 0 and C18 : 1ω9. The DNA G+C content of the isolate was 35.6 mol%. 16S rRNA gene sequence analysis showed that strain Z-1001T is a member of family Natranaerobiaceae , clustering with the type strain of Natranaerobius thermophilus (95.8–96.0 % sequence similarity). On the basis of physiological and phylogenetic data it is proposed that strain Z-1001T ( = DSM 24923T = VKM B-2666T) represents a novel genus and species, Natranaerobaculum magadiense gen. nov., sp. nov.
Three lactic acid bacteria were isolated from faeces of a jackal (Canis mesomelas) and raccoons (Procyron lotor). The isolates formed a subcluster in the Lactobacillus salivarius phylogenetic group, closely related to Lactobacillus animalis , Lactobacillus apodemi and Lactobacillus murinus , by phylogenetic analysis based on 16S rRNA and recA gene sequences. Levels of DNA–DNA relatedness revealed that the isolates belonged to the same taxon and were genetically separated from their phylogenetic relatives. The three strains were non-motile, obligately homofermentative and produced l-lactic acid as the main end-product from d-glucose. The strains metabolized raffinose. The major cellular fatty acids in the three strains were C16 : 0, C18 : 1ω9c and C19 : 1 cyclo 9,10. Based on the data provided, it is concluded that the three strains represent a novel species of the genus Lactobacillus , for which the name Lactobacillus faecis sp. nov. is proposed. The type strain is AFL13-2T ( = JCM 17300T = DSM 23956T).
A novel Gram-stain-positive, facultatively anaerobic, non-motile and lactic-acid-producing bacterium, designated strain ORL-24T, was isolated from the gut of the camel cricket, Diestrammena coreana. Optimal growth occurred at 37 °C, pH 8 and with 0 % (w/v) NaCl. The ratio of l-lactate to d-lactate in strain ORL-24T was 96 : 4. Lancefield antigen D was not detected. The strain was negative for oxidase activity and catalase activity. According to a phylogenetic analysis based on 16S rRNA gene sequences, strain ORL-24T was most closely related to the type strain of Enterococcus asini (96.9 % similarity). Comparative pheS and rpoA sequence analyses of strain ORL-24T indicated that the strain belonged to the genus Enterococcus . The major fatty acids were C16 : 0 and C18 : 1ω9c. The DNA G+C content was 41.3 mol%. Based on phenotypic, genotypic and phylogenetic analyses, strain ORL-24T represents a novel species of the genus Enterococcus , for which the name Enterococcus diestrammenae is proposed. The type strain is ORL-24T ( = KACC 16708T = JCM 18359T).
A novel filamentous bacterium, strain SCSIO 11153T, was isolated from a sediment sample collected from the Indian Ocean (80° 03.099′ E 01° 03.300′ N) at a depth of 4593 m. Good growth was observed at 50–55 °C and pH 7.0 with 3 % NaCl. It formed ivory–white colonies with radial wrinkles. Aerial mycelium was absent on the media tested. Phenotypic characteristics and 16S rRNA gene sequence analysis indicated that strain SCSIO 11153T belonged to the family Thermoactinomycetaceae . It exhibited 96.4 % and 96.2 % 16S rRNA gene sequence similarities to Melghirimyces algeriensis NariEXT and Melghirimyces thermohalophilus Nari11AT, respectively, while lower sequence similarity values (<95.4 %) were observed between strain SCSIO 11153T and other species of genera in the family Thermoactinomycetaceae . The menaquinone type was MK-7. Major cellular fatty acids were iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0. The polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine and phosphatidylglycerol. The DNA G+C content of strain SCSIO 11153T was 52.6 mol%. On the basis of the genotypic and phenotypic characteristics, it is proposed that strain SCSIO 11153T represents a novel species of the genus Melghirimyces with the name Melghirimyces profundicolus sp. nov. The type strain is SCSIO 11153T ( = DSM 45787T = CCTCC AA 2012007T = NBRC 109068T).
A novel Gram-stain-positive bacterium, designated strain YIM M11385T, was isolated from a marine sediment sample collected from the South Bay, Little Andaman Island, India with a salinity of 35 p.p.m., pH 8.5. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM M11385T belongs to the genus Marininema , supported by a bootstrap value of 100 %. The taxonomic position of this organism was further established by using a polyphasic approach. Strain YIM M11385T grew optimally at 28 °C, pH 7.0 and in the presence of 0–5 % (w/v) NaCl. The 16S rRNA gene sequence similarity between strain YIM M11385T and Marininema mesophilum SCSIO 10219T was 98.3 %. Strain YIM M11385T exhibited a quinone system with only MK-7, the polar lipid profile included diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as major components, and the major fatty acids were anteiso-C15 : 0, iso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The level of DNA–DNA relatedness between strain YIM M11385T and M. mesophilum SCSIO 10219T was 59.36 %. On the basis of genotypic and phenotypic data, it is apparent that strain YIM M11385T represents a novel species of the genus Marininema , for which the name Marininema halotolerans sp. nov. is proposed. The type strain is YIM M11385T ( = CCTCC AB 2012052T = DSM 45789T). In addition, we propose that the description of the genus Marininema should be further emended based on the results of the present study.
A Gram-positive, spore-forming, non-motile, strictly anaerobic rod-shaped bacterium was isolated from the caecal content of a TNFdeltaARE mouse. The isolate, referred to as strain SRB-521-5-IT, was originally cultured on a reduced agar medium containing yeast extract, rumen fluid and lactic acid as main energy and carbon sources. Phylogenetic analysis of partial 16S rRNA genes revealed that the species most closely related to strain SRB-521-5-IT were Flavonifractor plautii and Pseudoflavonifractor capillosus (<95 % sequence similarity; 1436 bp). In contrast to F. plautii and P. capillosus , strain SRB-521-5-IT contained a substantial amount of C18 : 0 dimethylacetal. Additional major fatty acids were C14 : 0 methyl ester, C16 : 0 dimethylacetal and C18 : 0 aldehyde. Strain SRB-521-5-IT differed in its enzyme profile from F. plautii and P. capillosus by being positive for dextrin, maltotriose, turanose, dl-lactic acid and d-lactic acid methyl ester but negative for d-fructose. In reduced Wilkins-Chalgren-Anaerobe broth, strain SRB-521-5-IT produced approximately 8 mM butyrate and 4 mM acetate. In contrast to F. plautii , the strain did not metabolize flavonoids. It showed intermediate resistance towards the antibiotics ciprofloxacin, colistin and tetracycline. Based on genotypic and phenotypic characteristics, we propose the name Intestinimonas butyriciproducens gen. nov., sp. nov. to accommodate strain SRB-521-5-IT ( = DSM 26588T = CCUG 63529T) as the type strain.
A Gram-reaction-positive, facultatively anaerobic, non-spore-forming and catalase-negative rod-shaped bacterial strain, designated IWT126T, was isolated from orchardgrass (Dactylis glomerata L.) silage preserved in Hachimantai, Iwate, Japan. The isolate showed growth at 15–45 °C, pH 3.5–7.5 and with 4.0 % (w/v) NaCl. The cell wall peptidoglycan did not contain meso-diaminopimelic acid, and the DNA G+C content was 45.6 mol%. The major cellular fatty acids were C16 : 0 and C19 : 1 cyclo 9,10. Based on 16S rRNA gene sequence similarity, strain IWT126T was classified as a member of the genus Lactobacillus and was most closely related to Lactobacillus odoratitofui YIT 11304T (98.7 %), Lactobacillus similis JCM 2765T (98.5 %), Lactobacillus collinoides JCM 1123T (97.6 %), Lactobacillus paracollinoides DSM 15502T (97.6 %) and Lactobacillus kimchicus DCY51T (96.9 %). Based on sequence analysis of the phenylalanyl-tRNA synthase α-subunit (pheS) gene, strain IWT126T was well separated from its phylogenetic neighbours in the genus Lactobacillus . Based on physiological, biochemical and genotypic results, as well as low DNA–DNA relatedness to recognized phylogenetic relatives in the genus Lactobacillus , classification of strain IWT126T as a representive of a novel species named Lactobacillus silagei sp. nov. is proposed. The type strain is IWT126T ( = JCM 19001T = DSM 27022T).
A coccal strain isolated from fresh broccoli was initially identified as Enterococcus saccharolyticus ; however, molecular identification and phenotypic traits did not support this identification. DNA–DNA hybridization with the type strain of E. saccharolyticus (76.4 % relatedness), DNA G+C content (35.7 mol%), phylogenetic analysis based on 16S rRNA, pheS and rpoA gene sequences, rep-PCR fingerprinting and profiles of cellular fatty acids, whole-cell proteins and enzyme activities, together with carbohydrate metabolism characteristics, indicated that this strain is distinct and represents a novel subspecies, for which the name Enterococcus saccharolyticus subsp. taiwanensis subsp. nov. is proposed. The type strain is 812T ( = NBRC 109476T = BCRC 80575T). Furthermore, we present an emended description of Enterococcus saccharolyticus and proposal of Enterococcus saccharolyticus subsp. saccharolyticus subsp. nov. (type strain ATCC 43076T = CCUG 27643T = CCUG 33311T = CIP 103246T = DSM 20726T = JCM 8734T = LMG 11427T = NBRC 100493T = NCIMB 702594T).
Three Gram-stain-positive bacterial strains, 11050T, 7-19T and 11102T, were isolated from traditional pickle and sourdough in Heilongjiang Province, China. These bacteria were characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, dnaK gene sequence analysis, fatty acid methyl ester (FAME) analysis, determination of DNA G+C content, DNA–DNA hybridization and an analysis of phenotypic features. Strain 11050T belonged to the Lactobacillus plantarum species group and shared 98.0–98.4 % 16S rRNA gene sequence similarities and 84.7–88.9 % dnaK gene sequence similarities with type strains of Lactobacillus plantarum subsp. plantarum , Lactobacillus plantarum subsp. argentoratensis , Lactobacillus pentosus , Lactobacillus paraplantarum , Lactobacillus fabifermentans and Lactobacillus xiangfangensis and had 75.9–80.7 % pheS gene sequence similarities and 90.7–92.5 % rpoA gene sequence similarities with Lactobacillus plantarum subsp. plantarum LMG 6907T, Lactobacillus plantarum subsp. argentoratensis LMG 9205, Lactobacillus pentosus LMG 10755T, Lactobacillus paraplantarum LMG 16673T, Lactobacillus fabifermentans LMG 24284T and Lactobacillus xiangfangensis 3.1.1T, respectively. Strain 7-19T was phylogenetically related to Lactobacillus thailandensis , Lactobacillus pantheris and Lactobacillus sharpeae , having 94.1–96.7 % 16S rRNA gene sequence similarities, 71.5–82.3 % pheS gene sequence similarities and 71.2–83.4 % rpoA gene sequence similarities with type strains of Lactobacillus thailandensis , Lactobacillus pantheris and Lactobacillus sharpeae , respectively. Strain 11102T was phylogenetically related to Lactobacillus oligofermentans , Lactobacillus suebicus , Lactobacillus vaccinostercus and Lactobacillus hokkaidonensis . Strain 11102T had 99.2 % 16S rRNA gene sequence similarity, 81.3 % pheS gene sequence similarity and 96.1 % rpoA gene sequence similarity with Lactobacillus oligofermentans LMG 22743T, respectively. Strain 11102T shared 96.0–96.8 % 16S rRNA gene sequence similarities, 73.3–81.0 % pheS gene sequence similarities and 74.6–76.9 % rpoA gene sequence similarities with type strains of Lactobacillus suebicus , Lactobacillus vaccinostercus and Lactobacillus hokkaidonensis , respectively. Based upon the data from polyphasic characterization obtained in the present study, three novel species, Lactobacillus mudanjiangensis sp. nov., Lactobacillus songhuajiangensis sp. nov. and Lactobacillus nenjiangensis sp. nov., are proposed and the type strains are 11050T ( = LMG 27194T = CCUG 62991T), 7-19T ( = LMG 27191T = NCIMB 14832T = CCUG 62990T) and 11102T ( = LMG 27192T = NCIMB 14833T), respectively.
A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterium capable of growing at 15–42 °C (optimum 30 °C) and at pH 5–11 (optimum pH 7) was isolated from compost. Its taxonomic position was deduced using a polyphasic approach and the strain was designated RC2T. 16S rRNA gene sequence analysis showed that the isolate belongs to the division Firmicutes , forming a clade within the cluster containing Bacillus flexus IFO 15715T, and showed highest similarity to B. flexus IFO 15715T (98.1 %). The cell wall contained meso-diaminopimelic acid as the diagnostic diamino acid. The major cellular fatty acids of the novel strain were iso-C15:0 (36.83 %), anteiso-C15:0 (49.19 %) and C16:0 (5.19 %). DNA–DNA hybridization between strain RC2T and B. flexus DSM 1320T showed a level of relatedness of 54.5 %. The polar lipid profile of strain RC2T showed the presence of phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine. The predominant isoprenoid quinone was MK-7 and the G+C content of strain RC2T was 37.6 mol%. On the basis of phenotypic characteristics, phylogenetic analysis and the results of biochemical and physiological tests, strain RC2T was clearly distinguished from closely related members of the genus, and the strain is assigned to a novel species, for which the name Bacillus paraflexus sp. nov. is proposed. The type strain is RC2T ( = MTCC 9831T = MCC 2100T = KCTC 13724T = CCM 7754T).