Ten Lactobacillus strains, previously isolated from different Bobal grape wines from the Utiel-Requena Origin Denomination of Spain, were characterized phylogenetically, genotypically and phenotypically. The 16S rRNA genes were sequenced and phylogenetic analysis showed that they form a tight phylogenetic clade that is closely related to reference strains Lactobacillus satsumensis NRIC 0604T, ‘Lactobacillus uvarum’ 8 and Lactobacillus mali DSM 20444T. DNA–DNA hybridization results confirmed the separation of the strains from other Lactobacillus species. Genotypically, the strains could be differentiated from their closest neighbours by 16S amplified rDNA restriction analysis and random amplified polymorphic DNA patterns. The strains were Gram-staining-positive, facultatively anaerobic rods that did not exhibit catalase activity. Phenotypically, they could be distinguished from their closest relatives by several traits such as their inabilities to grow at pH 3.3, to ferment sucrose, amygdalin and arbutin or to hydrolyse aesculin. The characteristics of the ten wine isolates suggest that they represent a novel species, for which the name Lactobacillus oeni sp. nov. is proposed. The type strain is 59bT (=CECT 7334T =DSM 19972T).
Two Gram-positive, rod-shaped, alkali-tolerant (pH 10.5), endospore-forming bacteria (strains KSC SF8bT and KSC SF10a) were isolated from surfaces within the Payload Hazardous Servicing Facility, where robotic spacecraft are assembled and tested before launch, at the Kennedy Space Center at Cape Canaveral. Based on 16S rRNA gene sequence similarities, these strains were shown to belong to the family Bacillaceae and the genus Bacillus. The highest 16S rRNA gene sequence similarity was ∼97.5 %, observed between the novel strains and Bacillus selenatarsenatis SF-1T. Several phenotypic characteristics, such as growth with 10 % NaCl and assimilation of melibiose and lactose, were useful in the discrimination of this novel species from the closely related alkali-tolerant species Bacillus firmus and B. selenatarsenatis. DNA–DNA hybridization studies revealed reassociation values of less than 45 % between strain KSC SF8bT and its closest genotypic neighbours. The combination of unique phenotypic and genotypic characteristics allowed the differentiation of these alkali- and halotolerant spore-forming strains from related Bacillus species, and a novel species, Bacillus canaveralius sp. nov., is proposed. The type strain is KSC SF8bT (=ATCC BAA-1493T=MTCC 8908T).
Novel strains of anaerobic, halophilic, alkalithermophilic bacteria were isolated from sediments of the alkaline, hypersaline lakes of the Wadi An Natrun, Egypt. Strains JW/NM-WN-LUT (and related strains) and JW/NM-WN-LH1T were non-spore-forming, non-motile bacteria with a Gram-type positive cell wall. Optimal growth of strain JW/NM-WN-LUT occurred at pH55 °C 9.5, 52 °C and with 3.7 M Na+ (2.2 M added NaCl). Optimal growth of strain JW/NM-WN-LH1T occurred at pH55 °C 9.9, 51 °C and with 3.9 M Na+ (2.3 M added NaCl). Both strains were obligately anaerobic and chemoorganotrophic; producing lactate and acetate as organic acids from pyruvate in different ratios. The DNA G+C contents of strains JW/NM-WN-LUT and JW/NM-WN-LH1T were 41.0 and 42.0 mol%, respectively. The main cellular fatty acids in both strains were anteiso-branched 15 : 0 and iso-branched 15 : 0. Phylogenetic analysis showed that both novel isolates belonged to the family Natranaerobiaceae within the order Natranaerobiales. Based on genotypic and phenotypic data, strain JW/NM-WN-LUT (=DSM 18760T=ATCC BAA-1443T) represents the type strain of a novel species, Natranaerobius trueperi sp. nov. Strain JW/NM-WN-LH1T represents a novel genus and species within the family Natranaerobiaceae, Natronovirga wadinatrunensis gen. nov., sp. nov. The type strain of the type species is JW/NM-WN-LH1T (=DSM 18770T=ATCC BAA-1444T).
A Gram-positive, moderately halophilic, alkalitolerant, strictly aerobic, oxidase- and catalase-positive, rod-shaped bacterium, strain YIM kkny3T, was isolated from a sediment sample collected from a salt lake in the Qaidam Basin of north-west China. Cells were motile by means of peritrichous flagella and formed ellipsoidal endospores lying in subterminal swollen sporangia. Growth occurred with 1–20 % (w/v) total salts (optimum, 5–10 %) and at pH 6.0–10.5 (optimum, pH 7.5–8.0) and 10–55 °C (optimum, 35–40 °C). It was unable to grow with NaCl as the only salt. meso-Diaminopimelic acid was present in the cell-wall peptidoglycan. The strain contained menaquinone 7 (MK-7) as the predominant respiratory quinone and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid as polar lipids. The major cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0. The DNA G+C content was 40.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM kkny3T belonged to the genus Virgibacillus, and was most closely related to the type strains of Virgibacillus olivae (97.1 % similarity), Virgibacillus marismortui (97.0 %) and Virgibacillus kekensis (96.8 %). Levels of DNA–DNA relatedness between strain YIM kkny3T and the type strains of V. olivae, V. marismortui and V. kekensis were 12.4, 10.6 and 15.7 %, respectively. The combination of phylogenetic analysis, genotypic data, phenotypic characteristics and chemotaxonomic differences indicated that strain YIM kkny3T represents a novel species of the genus Virgibacillus, for which the name Virgibacillus sediminis sp. nov. is proposed. The type strain is YIM kkny3T (=CCTCC AA 207023T=DSM 19797T=KCTC 13193T).
Strain FIRDI 006T, isolated from silage cattle feed, was characterized by using phenotypic and molecular taxonomic methods. The strain was a homofermentative lactic acid bacterium; the cells stained Gram-positive and were catalase-negative, non-motile, facultatively anaerobic rods. 16S rRNA gene sequence analysis revealed that the strain belongs phylogenetically to the genus Lactobacillus and can be placed within the Lactobacillus acidophilus–delbrueckii group. Lactobacillus gasseri and Lactobacillus johnsonii were the most closely related species, with 16S rRNA gene sequence similarities of 99.53 and 99.46 % to the respective type strains. Low gyrB gene sequence similarities (<90 %) and low DNA–DNA reassociation values (<45 %) were obtained between the strain and the phylogenetically closest neighbours. Based on phenotypic and genetic evidence, the strain is considered to represent a novel species, for which the name Lactobacillus taiwanensis sp. nov. is proposed. The type strain is FIRDI 006T (=BCRC 17755T =DSM 21401T).
Two Gram-variable, aerobic, motile, rod-shaped, endospore-forming bacterial strains, M4-2T and M4-1, were isolated from soil samples collected from Oblast Magadan, Russian Far East, as micro-organisms antagonistic to the psychrophilic phytopathogenic fungus Typhula ishikariensis. Strains M4-2T and M4-1 were identified as members of the genus Paenibacillus by phenotypic and phylogenetic analyses based on 16S rRNA gene sequences. The strains contained anteiso-C15 : 0 as the major fatty acid (63.0–64.7 %) and MK-7 as the major isoprenoid quinone. The DNA G+C contents were 42.8 and 41.7 mol%, respectively. 16S rRNA gene sequence analysis showed that strains M4-2T and M4-1 exhibited high similarities with Paenibacillus macquariensis DSM 2T (99.5 and 99.7 %, respectively) and Paenibacillus antarcticus LMG 22078T (99.4 and 99.5 %, respectively). There were no clear differences in the phenotypic characteristics and chemotaxonomic and phylogenetic data between the novel isolates and P. macquariensis DSM 2T. DNA–DNA hybridization experiments between strain M4-2T and P. macquariensis DSM 2T and P. antarcticus LMG 22078T revealed reassociation values of 56 and 49 %, respectively. Multilocus sequence analysis confirmed the differences between the new isolates and reference strains that were observed with the DNA–DNA hybridization studies. On the basis of the results described above, it is proposed that the isolates represent a novel subspecies of P. macquariensis, Paenibacillus macquariensis subsp. defensor subsp. nov. The type strain is M4-2T (=JCM 14954T=NCIMB 14397T).
Two strictly anaerobic, crescent-shaped bacterial strains, designated WGT and Ycb08, were isolated from a cellulose-degrading mixed culture enriched from yak rumen contents. The strains were Gram-negative, non-spore-forming and motile, with four to six flagella situated at the centre of the concave side of the cell. The cells were 0.9–1.1×4–6 μm. Growth was observed at 27–46 °C (optimum 39 °C) and pH 4.2–8.3 (optimum pH 7.0–7.2). Arabinose, glucose, mannose, cellobiose, lactose, sucrose, trehalose, melibiose, raffinose, salicin and aesculin were fermented. The end products of glucose fermentation were acetate, propionate and CO2. The G+C contents of strains WGT and Ycb08 were respectively 63.9±0.2 and 62.5±0.2 mol% (T m). Phylogenetic analysis based on 16S rRNA gene sequences revealed that the two strains were related to the genera Mitsuokella and Selenomonas at similarity levels below 97 %; however, they differed from members of the genus Mitsuokella in their flagellar arrangement. On the basis of phenotypic, genotypic and physiological evidence, strains WGT and Ycb08 are identified as members of a novel species of the genus Selenomonas, for which the name Selenomonas bovis sp. nov. is proposed. The type strain is WGT (=CGMCC 1.5073T =JCM 15470T).