Pullulanibacillus camelliae sp. nov., isolated from Pu'er tea.

A novel Gram-stain-positive, strictly aerobic, endospore-forming, rod-shaped bacterial strain 7578-24T was isolated from ripened Pu'er tea. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 7578-24T clustered with species of the genus Pullulanibacillus in the family Sporolactobacillaceae with 97.8-95.2 % sequence similarities, and was most closely related to Pullulanibacillus pueri YN3T with 97.8 % 16S rRNA gene sequence similarity. The DNA-DNA relatedness value between strain 7578-24T and P. pueri YN3T was 35 %. Strain 7578-24T had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The major menaquinone was menaquinone 7 (MK-7). C18 : 1ω7c (45.4 %), anteiso-C17 : 0 (30.6 %) and anteiso-C15 : 0 (10.1 %) were the predominant fatty acids, and diphosphatidylglycerol, phosphatidylglycerol, five unknown phospholipids and one unknown aminolipid were the major polar lipids. The DNA G+C content of strain 7578-24T was 45.2 mol%. Strain 7578-24T could be differentiated from other related species of the genus Pullulanibacillus based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of polyphasic evidence from this study, a novel species of the genus Pullulanibacillus named Pullulanibacillus camelliae sp. nov. is proposed, with strain 7578-24T (=CGMCC 1.15371T=JCM 31236T) as the type strain.

Cohnella saccharovorans sp. nov., isolated from ginseng soil.

A novel bacterial strain, CJ22T, was isolated from soil of a ginseng field located in Anseong, Korea. Cells of strain CJ22T were aerobic, Gram-stain-positive, endospore-forming, motile, oxidase- and catalase-positive and rod-shaped. The isolate grew optimally at pH 7 and 30 °C. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CJ22T belonged to the genus Cohnella, displaying highest sequence similarity of 97.3% with Cohnella panacarvi Gsoil 349T. DNA-DNA relatedness between strain CJ22T and its closest relative was 35.5 % (reciprocal value, 23.8%). The phenotypic features of strain CJ22T also distinguished it from related species of the genus Cohnella. The diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. The major isoprenoid quinone was menaquinone MK-7 and the major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, lysyl-phosphatidylglycerol, two unidentified phospholipids and two unidentified aminophospholipids. The predominant cellular fatty acids of strain CJ22T were anteiso-C15 : 0, iso-C16:0 and C16:0. The DNA G+C content was 63.1 mol%. Based on data from this polyphasic taxonomic study, strain CJ22T is considered to represent a novel species of the genus Cohnella, for which the name Cohnella saccharovorans sp. nov. is proposed. The type strain is CJ22T (=KACC 17501T=JCM 19227T).

Pullulanibacillus pueri sp. nov., isolated from Pu'er tea.

A novel Gram-stain-positive, aerobic, endospore-forming, rod-shaped bacterial strain YN3(T) was isolated from ripened Pu'er tea. Phylogenetic analysis of 16S rRNA gene sequences showed that the strain belonged to the family Sporolactobacillaceae and was closely related to Pullulanibacillus naganoensis DSM 10191(T) (95.8% 16S rRNA gene sequence similarity) and Pullulanibacillus uraniitolerans DSM 19429(T) (95.4%). Growth of the strain was observed at 20-50 °C (optimum 30-37 °C), at pH 4.0-8.0 (optimum pH 5.0-6.0). The strain had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinone was menaquinone-7 (MK-7). The major fatty acids were anteiso-C15:0, anteiso-C17:0 and C18:1ω7c. The DNA G+C content of strain YN3(T) was 38.7 mol%. Strain YN3(T) could be differentiated from recognized species of the genus Pullulanibacillus based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of polyphasic evidence from this study, Pullulanibacilluspueri sp. nov., is proposed, with strain YN3(T) ( = CGMCC 1.12777(T ) = JCM 30075(T)) as the type strain.

Fontibacillus solani sp. nov. isolated from potato (Solanum tuberosum L.) root.

A bacterial strain designated A4STR04(T) was isolated from the inner root tissue of potatoes in Spain. Phylogenetic analysis based on the 16S rRNA gene sequence placed the isolate into the genus Fontibacillus, being most closely related to Fontibacillus panacisegetis KCTC 13564(T) with 99% identity. The isolate was observed to form Gram-positive, motile and sporulating rods. The catalase test was found to be negative and oxidase positive. Nitrate was found to be reduced to nitrite. ß-Galactosidase and caseinase were observed to be produced but the production of gelatinase, urease, arginine dehydrolase, ornithine and lysine decarboxylase was negative. Aesculin hydrolysis was found to be positive and acetoin production was negative. Growth was found to be supported by many carbohydrates and organic acids as carbon source. MK-7 was the only menaquinone detected and the major fatty acid (61.5%) was identified as anteiso-C(15:0), as occurs in the other species of genus Fontibacillus. The strain A4STR04(T) was found to display a complex lipid profile consisting of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, a glycolipid, two phospholipids, a lipid and two aminophospholipids. Mesodiaminopimelic acid was detected in the peptidoglycan. The G+C content was determined to be 50.5 mol% (Tm). Phylogenetic, chemotaxonomic and phenotypic analyses showed that strain A4STR04(T) (=LMG 28458 (T) = CECT 8693(T)) should be classified as representing a novel species of genus Fontibacillus, for which the name Fontibacillus solani sp. nov. is proposed.

Biodegradation of complex hydrocarbons in spent engine oil by novel bacterial consortium isolated from deep sea sediment.

Complex hydrocarbon and aromatic compounds degrading marine bacterial strains were isolated from deep sea sediment after enrichment on spent engine (SE) oil. Phenotypic characterization and phylogenetic analysis of 16S rRNA gene sequences showed the isolates were related to members of the Pseudoalteromonas sp., Ruegeria sp., Exiguobacterium sp. and Acinetobacter sp. Biodegradation using 1% (v/v) SE oil with individual and mixed strains showed the efficacy of SE oil utilization within a short retention time. The addition of non-ionic surfactant 0.05% (v/v) Tween 80 as emulsifying agent enhanced the solubility of hydrocarbons and renders them more accessible for biodegradation. The degradation of several compounds and the metabolites formed during the microbial oxidation process were confirmed by Fourier transform infrared spectroscopy and Gas chromatography-mass spectrometry analyses. The potential of this consortium to biodegrade SE oil with and without emulsifying agent provides possible application in bioremediation of oil contaminated marine environment.

Identification and characterization of novel catalytic bioscavengers of organophosphorus nerve agents.

In an effort to discover novel catalytic bioscavengers of organophosphorus (OP) nerve agents, cell lysates from a diverse set of bacterial strains were screened for their capacity to hydrolyze the OP nerve agents VX, VR, and soman (GD). The library of bacterial strains was identified using both random and rational approaches. Specifically, two representative strains from eight categories of extremophiles were chosen at random. For the rational approach, the protein sequence of organophosphorus hydrolase (OPH) from Brevundimonas diminuta was searched against a non-redundant protein database using the Basic Local Alignment Search Tool to find regions of local similarity between sequences. Over 15 protein sequences with significant sequence similarity to OPH were identified from a variety of bacterial strains. Some of these matches were based on predicted protein structures derived from bacterial genome sequences rather than from bona fide proteins isolated from bacteria. Of the 25 strains selected for nerve agent testing, three bacterial strains had measurable levels of OP hydrolase activity. These strains are Ammoniphilus oxalaticus, Haloarcula sp., and Micromonospora aurantiaca. Lysates from A. oxalaticus had detectable hydrolysis of VR; Haloarcula sp. had appreciable hydrolysis of VX and VR, whereas lysates from M. aurantiaca had detectable hydrolysis of VR and GD.

Exiguobacterium oxidotolerans, a halotolerant plant growth promoting rhizobacteria, improves yield and content of secondary metabolites in Bacopa monnieri (L.) Pennell under primary and secondary salt stress.

Brahmi (Bacopa monnieri), an integral component of Indian Ayurvedic medicine system, is facing a threat of extinction owing to the depletion of its natural populations. The present study investigates the prospective of exploitation of halotolerant plant growth promoting rhizobacteria (PGPR) in utilising the salt stressed soils for cultivation of B. monnieri. The effects of two salt tolerant PGPR, Bacillus pumilus (STR2) and Exiguobacterium oxidotolerans (STR36) on the growth and content of bacoside-A, an important pharmaceutical compound in B. monnieri, were investigated under primary and secondary salinity conditions. The herb yields of un-inoculated plants decreased by 48 % under secondary salinization and 60 % under primary salinization than the non salinised plants. Among the rhizobacteria treated plants, E. oxidotolerans recorded 109 and 138 %, higher herb yield than non-inoculated plants subjected to primary and secondary salinity respectively. E. oxidotolerans inoculated plants recorded 36 and 76 % higher bacoside-A content under primary and secondary salinity respectively. Higher levels of proline content and considerably lower levels of lipid peroxidation were noticed when the plants were inoculated with PGPR under all salinity regimes. From the results of this investigation, it can be concluded that, the treatments with salt tolerant PGPR can be a useful strategy in the enhancement of biomass yield and saponin contents in B. monnieri, as besides being an eco-friendly approach; it can also be instrumental in cultivation of B. monnieri in salt stressed environments.

Fontibacillus panacisegetis sp. nov., isolated from soil of a ginseng field.

A facultatively anaerobic, endospore-forming bacterium, designated strain P11-6(T), was isolated from soil of a ginseng field located in Geumsan County, Republic of Korea. Cells of strain P11-6(T) were Gram-stain-negative, catalase-negative, motile rods and produced semi-translucent, circular, white colonies on tryptic soy agar. The isolate contained MK-7 as the only menaquinone and anteiso-C(15 : 0) as the major fatty acid. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unknown aminophosphoglycolipid, an unknown aminophospholipid, two unknown phospholipids, three unknown glycolipids and three unknown lipids were detected in the polar lipid profile. The DNA G+C content of strain P11-6(T) was 41.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequencing showed that strain P11-6(T) was most closely related to Fontibacillus aquaticus GPTSA 19(T) (97.2 % sequence similarity) and that it formed a separate lineage with F. aquaticus in the family Paenibacillaceae. Combined phenotypic and DNA-DNA hybridization data supported the conclusion that strain P11-6(T) represents a novel species in the genus Fontibacillus, for which the name Fontibacillus panacisegetis sp. nov. is proposed; the type strain is P11-6(T) (=KCTC 13564(T) =CECT 7605(T)).

Cohnella yongneupensis sp. nov. and Cohnella ginsengisoli sp. nov., isolated from two different soils.

Two aerobic, Gram-positive, rod-shaped bacterial strains, 5YN10-14(T) and GR21-5(T), were isolated from the Yongneup wetland and ginseng soil in Korea, respectively. The two strains formed ellipsoidal or oval spores positioned centrally or paracentrally in swollen sporangia. On the basis of 16S rRNA gene sequence analysis, these strains were related to members of the genus Cohnella. 16S rRNA gene sequence similarity between strains 5YN10-14(T) and GR21-5(T) was 95.9 %. Strains 5YN10-14(T) and GR21-5(T) showed, respectively, 94.3 and 95.2 % 16S rRNA gene sequence similarity to Cohnella thermotolerans CCUG 47242(T), 94.6 and 94.4 % to Cohnella hongkongensis HKU3(T), 94.7 and 94.7 % to Cohnella laeviribosi RI-39(T), and 95.4 and 94.8 % to Cohnella phaseoli GSPC1(T). The major fatty acids of strain 5YN10-14(T) were anteiso-C(15 : 0) (51.1 %), iso-C(16 : 0) (18.5 %) and C(16 : 0) (13.2 %), and the major fatty acids of strain GR21-5(T) were anteiso-C(15 : 0 ) (48.9 %), iso-C(16 : 0) (15.0 %) and iso-C(15 : 0) (12.2 %). The two strains contained menaquinone with seven isoprene units (MK-7) as the predominant quinone, and diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine as major polar lipids; however, strain 5YN10-14(T) also contained lysylphosphatidylglycerol as a major polar lipid, whereas strain GR21-5(T) had an unknown aminophospholipid as another major polar lipid. The DNA G+C contents of strains 5YN10-14(T) and GR21-5(T) were 58.8 and 61.3 mol%, respectively. Based on the results of the phylogenetic and phenotypic data presented, it was concluded that the two strains represent two novel species of the genus Cohnella , for which the names Cohnella yongneupensis sp. nov. (type strain 5YN10-14(T)=KACC 11768(T)=DSM 18998(T)) and Cohnella ginsengisoli sp. nov. (type strain GR21-5(T)=KACC 11771(T)=DSM 18997(T)) are proposed.