<Emphasis Type="Italic">Calculibacillus koreensis</Emphasis> gen. nov., sp. nov., an anaerobic Fe(III)-reducing bacterium isolated from sediment of mine tailings

A strictly anaerobic bacterium, strain B5^T, was isolated from sediment of an abandoned coal mine in Taebaek, Republic of Korea. Cells of strain B5^T were non-spore-forming, straight, Gram-positive rods. The optimum pH and temperature for growth were pH 7.0 and 30°C, respectively, while the strain was able to grow within pH and temperature ranges of 5.5–7.5 and 25–45°C, respectively. Growth of strain B5^T was observed at NaCl concentrations of 0 to 6.0% (w/v) with an optimum at 3.0–4.0% (w/v). The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, an unknown phospholipid and three unknown polar lipids. Strain B5^T grew anaerobically by reducing nitrate, nitrite, ferric-citrate, ferric-nitrilotriacetate, elemental sulfur, thiosulfate, and anthraquinone-2-sulfonate in the presence of proteinaceous compounds, organic acids, and carbohydrates as electron donors. The isolate was not able to grow by fermentation. Strain B5^T did not grow under aerobic or microaerobic conditions. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain B5^T is most closely related to the genus Tepidibacillus (T. fermentans STGH^T; 96.3%) and Vulcanibacillus (V. modesticaldus BR^T; 94.6%). The genomic DNA G+C content (36.9 mol%) of strain B5^T was higher than those of T. fermentans STGH^T (34.8 mol%) and V. modesticaldus BR^T (34.5 mol%). Based on its phenotypic, chemotaxonomic, and phylogenetic properties, we describe a new species of a novel genus Calculibacillus, represented by strain B5^T (=KCTC 15397^T =JCM 19989^T), for which we propose the name Calculibacillus koreensis gen. nov., sp. nov.     

Role of a Ferric Ion-Reducing System in Sulfur Oxidation of Thiobacillus ferrooxidans

The properties of a ferric ion-reducing system which catalyzes the reduction of ferric ion with elemental sulfur was investigated with a pure strain of Thiobacillus ferrooxidans. In anaerobic conditions, washed intact cells of the strain reduced 6 mol of Fe³⁺ with 1 mol of elemental sulfur to give 6 mol of Fe²⁺, 1 mol of sulfate, and a small amount of sulfite. In aerobic conditions, the 6 mol of Fe²⁺ produced was immediately reoxidized by the iron oxidase of the cell, with a consumption of 1.5 mol of oxygen. As a result, Fe²⁺ production was never observed under aerobic conditions. However, in the presence of 5 mM cyanide, which completely inhibits the iron oxidase of the cell, an amount of Fe²⁺ production comparable to that formed under anaerobic conditions was observed under aerobic conditions. The ferric ion-reducing system had a pH optimum between 2.0 and 3.8, and the activity was completely destroyed by 10 min of incubation at 60°C. A short treatment of the strain with 0.5% phenol completely destroyed the ferric ion-reducing system of the cell. However, this treatment did not affect the iron oxidase of the cell. Since a concomitant complete loss of the activity of sulfur oxidation by molecular oxygen was observed in 0.5% phenol-treated cells, it was concluded that the ferric ion-reducing system plays an important role in the sulfur oxidation activity of this strain, and a new sulfur-oxidizing route is proposed for T. ferrooxidans.     

<Emphasis Type="Italic">Methylobacillus caricis</Emphasis> sp. nov., an obligate methylotroph isolated from roots of sedge (<Emphasis Type="Italic">Carex</Emphasis> sp.)

A new obligately methylotrophic bacterium, strain OV^T, was isolated from roots of sedge (Carex sp.). The isolate was represented by aerobic gram-negative motile, non-spore-forming rods, which divided by binary fission. Optimal growth occurred at 22?29°C and pH 7.5?8.5 in the presence of 0.5?2% NaCl; growth was inhibited by 3.5% NaCl. Strain OV^T utilized methanol as the only carbon and energy source. The organism used the KDPG variant of the ribulose monophosphate pathway (RuMP) of С₁ metabolism. Ammonium was assimilated by reductive amination of α-ketoglutarate. The major cellular fatty acids were C_16:0 (45.5%), C_16:1ω7c (40.7%), and C_17cyc (8.0%). The major ubiquinone was Q₈. The dominant phospholipids were phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. The DNA G + C content of strain OV^T was 51.4 mol % (T_m). While the 16S rRNA gene sequence of strain OV^T exhibited high similarity to those of Methylobacillus species: M. gramineus Lap^T (99.6%) and M. glycogenes TK 0113^T (98.7%), the DNA-DNA hybridization level between strain OV^T and M. gramineus Lap^T was only 52%. Based on the data obtained, strain OV^T was assigned to the new species Methylobacillus caricis sp. nov. (=VKM B-3158^T = JCM 32031^T).     

<Emphasis Type="Italic">Thermosulfurimonas marina</Emphasis> sp. nov., an Autotrophic Sulfur-Disproportionating and Nitrate-Reducing Bacterium Isolated from a Shallow-Sea Hydrothermal Vent

A thermophilic, anaerobic, chemolithoautotrophic bacterium (strain SU872^T) was isolated from a shallow-sea hydrothermal vent at Kunashir Island. The cells were motile, gram-negative, oval or rodshaped 0.5?0.6 μm thick and 1.5?2.0 μm long, occurring singly or in pairs. Strain SU872^T grew at 50 to 79°C (optimum at 74°C), pH from 5.0 to 8.0 (optimum at 6.7?7.0), and NaCl concentration of 1.5–4.5%. Strain SU872^T was able to grow by disproportionation of elemental sulfur, thiosulfate, or sulfite, with CO₂/HCO₃^? as the sole carbon source. Growth was enhanced in the presence of ferrihydrite (poorly crystalline Fe(III) oxide) as as a sulfide-scavenging agent. Sulfate was not used as an electron acceptor. Growth also occurred with elemental sulfur, thiosulfate, or sulfite (but not sulfide) as electron donors and nitrate as an electron acceptor, with production of sulfate and ammonium. Analysis of the 16S rRNA gene sequence revealed 97.8% similarity between strain SU872^T and the type strain Thermosulfurimonas dismutans S95^T (phylum Thermodesulfobacteria). According to the results of DNA–DNA hybridization, the similarity of genomic DNA of the strains SU872^T and T. dismutans S95^T was 48%. Based on its phenotypic characteristics and the results of phylogenetic analysis, it is proposed to assign the isolate to a new species of the genus Thermosulfurimonas,—Thermosulfurimonas marina sp. nov., with the type strain SU872^T (=DSM 104922^T, =VKM B-3177^T, =UNIQEM SU872^T).     

Biogeochemistry of an Iron-Rich Hypersaline Microbial Mat (Camargue, France)

In situ microsensor measurements were combined with biogeochemical methods to determine oxygen, sulfur, and carbon cycling in microbial mats growing in a solar saltern (Salin-de-Giraud, France). Sulfate reduction rates closely followed the daily temperature changes and were highest during the day at 25°C and lowest during the night at 11°C, most probably fueled by direct substrate interactions between cyanobacteria and sulfate-reducing bacteria. Sulfate reduction was the major mineralization process during the night and the contribution of aerobic respiration to nighttime DIC production decreased. This decrease of aerobic respiration led to an increasing contribution of sulfide (and iron) oxidation to nighttime O₂ consumption. A peak of elemental sulfur in a layer of high sulfate reduction at low sulfide concentration underneath the oxic zone indicated anoxygenic photosynthesis and/or sulfide oxidation by iron, which strongly contributed to sulfide consumption. We found a significant internal carbon cycling in the mat, and sulfate reduction directly supplied DIC for photosynthesis. The mats were characterized by a high iron content of 56 mol Fe cm^–3, and iron cycling strongly controlled the sulfur cycle in the mat. This included sulfide precipitation resulting in high FeS contents with depth, and reactions of iron oxides with sulfide, especially after sunset, leading to a pronounced gap between oxygen and sulfide gradients and an unusual persistence of a pH peak in the uppermost mat layer until midnight.     

Physiologic Versatility and Growth Flexibility as the Main Characteristics of a Novel Thermoacidophilic Acidianus Strain Isolated from Copahue Geothermal Area in Argentina

A novel thermoacidophilic archaeal strain has been isolated from three geothermal acidic hot springs in Copahue, Argentina. One of the most striking characteristic of ALE1 isolate is its metabolic versatility. It grows on sulphur, tetrathionate, iron (II) and sucrose under aerobic conditions, but it can also develop under anaerobic conditions using iron (III) or sulphur as electron acceptors and sulphur or hydrogen as electron donors autotrophically. A temperature of 75 °C and a pH between 2.5 and 3.0 are strain ALE1 optimal growth conditions, but it is able to oxidise iron (II) even at pH 1.0. Cells are irregular cocci surrounded by a regularly arrayed glycoprotein layer (S-layer). Phylogenetic analysis shows that strain ALE1 belongs to the family Sulfolobaceae in the class Thermoprotei, within the phylum Crenarchaeota. Based on 16S rRNA gene sequence similarity on NCBI database, ALE1 does not have closely related relatives, neither in culture nor uncultured, which is more surprising. Its closest related species are strains of Acidianus hospitalis (91 % of sequence similarity), Acidianus infernus (90 %), Acidianus ambivalens (90 %) and Acidianus manzanensis (90 %). Its DNA base composition of 34.5 %?mol C?+?G is higher than that reported for other Acidianus species. Considering physiological and phylogenetic characteristics of strain ALE1, we considered it to represent a novel species of the genus Acidianus (candidatus “Acidianus copahuensis”). The aim of this study is to physiologically characterise this novel archaea in order to understand its role in iron and sulphur geochemical cycles in the Copahue geothermal area and to evaluate its potential applications in bioleaching and biooxidation.     

Desulfacinum subterraneumsp. nov., a New Thermophilic Sulfate-Reducing Bacterium Isolated from a High-Temperature Oil Field

A new thermophilic sulfate-reducing bacterium isolated from the high-temperature White Tiger oil field (Vietnam) is described. Cells of the bacterium are oval (0.4–0.6 by 0.6–1.8 m), nonmotile, non-spore-forming, and gram-negative. Growth occurs at 45 to 65°C (with an optimum at 60°C) at NaCl concentrations of 0 to 50 g/l. In the course of sulfate reduction, the organism can utilize lactate, pyruvate, malate, fumarate, ethanol, salts of fatty acids (formate, acetate, propionate, butyrate, caproate, palmitate), yeast extract, alanine, serine, cysteine, and H₂+ CO₂(autotrophically). In addition to sulfate, the bacterium can use sulfite, thiosulfate, and elemental sulfur as electron acceptors. In the absence of electron acceptors, the bacterium can ferment pyruvate and yeast extract (a yet unrecognized capacity of sulfate reducers) with the formation of acetate and H₂. The G+C content of DNA is 60.8 mol %. The level of DNA–DNA hybridization of the isolate (strain 101^T) and Desulfacinum infernum(strain BG1^T) is as low as 34%. Analysis of the nucleotide sequence of 16S rDNA places strain 101^Tin the phylogenetic cluster of the Desulfacinumspecies within the sulfate reducer subdivision of the delta subclass of Proteobacteria. All these results allowed the bacterium studied to be described as a new species, Desulfacinum subterraneumsp. nov., with strain 101 as the type strain.     

<Emphasis Type="Italic">Spirosoma lituiforme</Emphasis> sp. nov., isolated from soil

A Gram-staining-negative, non-motile, curved rod-shaped, aerobic bacterium, designated S1-2-4^T, was isolated from soil in Jeollabuk-do province, Republic of Korea, and was characterized taxonomically using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain S1-2-4^T was a member of the family Cytophagaceae and most closely related to ‘Spirosoma radiotolerans’ DG5A (97.2%), Spirosoma fluviale MSd3^T (96.4%), and Spirosoma linguale DSM 74^T (96.3%). The genomic DNA G + C content of strain S1-2-4^T was 49.7 mol%. The major fatty acids were summed feature 3 (C_16:1ω7c/C_16:1ω6c), C_16:1ω5c, and C_16:0, and the major polar lipid was phosphatidylethanolamine. MK-7 was the predominant respiratory quinone. Phenotypic and chemotaxonomic data supported the affiliation of strain S1-2-4^T with the genus Spirosoma. DNA-DNA hybridization between strain S1-2-4^T and ‘Spirosoma radiotolerans’ showed relatively low DNA-DNA relatedness (31%). Strain S1-2-4^T could be distinguished from its closest phylogenetic neighbors based on its phenotypic, genotypic, and chemotaxonomic features. Therefore, strain S1-2-4^T represents a novel member of the genus Spirosoma, for which the name Spirosoma lituiforme sp. nov. is proposed. The type strain is S1-2-4^T (= KCTC 52724^T = JCM 32128^T).     

<Emphasis Type="Italic">Hymenobacter terrigena</Emphasis> sp. nov., isolated from soil

A Gram-stain-negative, non-motile, non-spore-forming, rodshaped, aerobic bacterial strain, designated S1-2-2-5^T, was isolated from the Jeollabuk-do province, Republic of Korea, and was characterized taxonomically using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain S1-2-2-5^T belonged to the family Cytophagaceae in phylum Bacteroidetes, and was most closely related to Hymenobacter terrae DG7A^T (98.2%), Hymenobacter rubidus DG7B^T (98.0%), Hymenobacter soli PB17^T (97.7%), Hymenobacter daeguensis 16F3Y-2^T (97.2%) and Hymenobacter saemangeumensis GSR0100^T (97.0%). The G + C content of the genomic DNA of strain S1-2-2-5^T was 59.4 mol%. The detection of menaquinone MK-7 as the predominant respiratory quinone, a fatty acid profile with summed feature 3 (C_16:1ω7c/C_16:1ω6c; 32.0%), C_15:0 iso (19.0%), and C_15:0 anteiso (15.0%) as the major components, and a polar lipid profile with phosphatidylethanolamine as the major component supported the affiliation of strain S1-2-2-5^T to the genus Hymenobacter. The DNA-DNA relatedness between strain S1-2-2-5^T and H. terrae KCTC 32554^T, H. rubidus KCTC 32553^T, H. soli KCTC 12607^T, H. daeguensis KCTC 52537^T, and H. saemangeumensis KACC 16452^T were 49.5, 48.2, 34.1, 28.1, and 31.8% respectively, clearly showing that the isolate is not related to them at the species level. Strain S1-2-2-5^T could be clearly differentiated from its closest neighbors on the basis of its phenotypic, genotypic and chemotaxonomic features. Therefore, strain S1-2-2-5^T represents a novel species of the genus Hymenobacter, for which the name Hymenobacter terrigena sp. nov. is proposed. The type strain is S1-2-2-5^T (= KCTC 52737^T = JCM 32195^T).     

<Emphasis Type="Italic">Thialkalivibrio halophilus</Emphasis> sp. nov., a novel obligately chemolithoautotrophic,facultatively alkaliphilic,and extremely salt-tolerant,sulfur-oxidizing bacterium from a hypersaline alkaline lake

A new chemolithoautotrophic, facultatively alkaliphilic, extremely salt-tolerant, sulfur-oxidizing bacterium was isolated from an alkaline hypersaline lake in the Altai Steppe (Siberia, Russia). According to 16S rDNA analysis and DNA–DNA hybridization, strain HL 17^T was identified as a new species of the genus Thialkalivibrio belonging to the subdivision of the Proteobacteria for which the name Thialkalivibrio halophilus is proposed. Strain HL 17^T is an extremely salt-tolerant bacterium growing at sodium concentrations between 0.2 and 5 M, with an optimum of 2 M Na⁺. It grew at high concentrations of NaCl and of Na₂CO₃/NaHCO₃ (soda). Strain HL 17^T is a facultative alkaliphile growing at pH range 7.5–9.8, with a broad optimum between pH 8.0 and 9.0. It used reduced inorganic sulfur compounds (thiosulfate, sulfide, polysulfide, elemental sulfur, and tetrathionate) as energy sources and electron donors. In continuous culture under energy limitation, thiosulfate was stoichiometrically oxidized to sulfate. In sodium carbonate medium under alkaline conditions, the maximum growth rate was similar, while the biomass yield was lower as compared with the NaCl-grown culture. The maximum sulfur-oxidizing capacity measured in washed cells was higher in the soda buffer independent of the growth conditions. The compatible solute content of the biomass was higher in the sodium chloride-grown culture than in the sodium carbonate/bicarbonate-grown culture. The data suggest that the osmotic pressure differences between soda and NaCl solutions might be responsible for the difference observed in compatible solutes production. This may have important implications in overall energetic metabolism of high salt adaptation.     

<Emphasis Type="Italic">Sphingorhabdus buctiana</Emphasis> sp. nov., isolated from fresh water,and reclassification of <Emphasis Type="Italic">Sphingopyxis contaminans</Emphasis> as <Emphasis Type="Italic">Sphingorhabdus contaminans</Emphasis> comb. nov.

A Gram-stain-negative, strictly aerobic, non-motile and rod-shaped bacterial strain, designated T5^T, was isolated from the Chishui River in Maotai town, Guizhou Province, Southwest of China. Strain T5^T was found to grow optimally at pH 9.0 and 25 °C. The 16S rRNA gene sequence analysis indicated that strain T5^T belongs to the family Sphingomonadaceae within the phylum Proteobacteria; the strain T5^T clustered with the type strains of Sphingopyxis contaminans, Sphingorhabdus wooponensis and Sphingorhabdus rigui, with which it exhibits 16S rRNA gene sequence similarity values of 96.2–96.9%. The DNA G+C content was 58.5 mol%. The major respiratory quinone was Q-10 and the major polar lipid was phosphatidylethanolamine. The major polyamine was homospermidine and the major fatty acids were C_18:1 ω7c (37.5%) and C_16:1 ω7c (30.1%). On the basis of phylogenetic, phenotypic and genetic data, strain T5^T represents a novel species of the genus Sphingorhabdus, for which the name Sphingorhabdus buctiana sp. nov. is proposed. The type strain is T5^T (= CGMCC 1.12929^T = JCM 30114^T). It is also proposed that Sphingopyxis contaminans should be reclassified as a member of the genus Sphingorhabdus.     

<Emphasis Type="Italic">Methylophaga muralis</Emphasis> Bur 1, a haloalkaliphilic methylotroph isolated from the Khilganta soda lake (Southern Transbaikalia,Buryat Republic)

A haloalkaliphilic restricted facultative methylotroph, strain Bur 1, which used methanol, methylated amines, and fructose as carbon and energy sources, was isolated from the soda Lake Khilganta (Buryat Republic, Russia). The cells were gram-negative non-spore-forming, motile rods reproducing by binary fission. The organism was aerobic, reduced nitrates to nitrites. Growth occurred at temperatures from 4 to 37°C (optimum at 25–29°C), pH 7.5–10.5 (optimum at 8.5–9.5), and NaCl concentration in the medium from 0.05 to 10.0% NaCl (optimum at 3–4%). Ectoine, glutamate, and sucrose were accumulated as osmoprotectants. Activity of the enzymes of de novo ectoine biosynthesis were detected. The organism utilized C₁ compounds via the KDPG variant of the ribulose monophosphate pathway. The DNA G + C content was 44.67 mol %. Based on the similarity of the 16S rRNA gene sequences (94.7–99.1%) and the results of DNA–DNA hybridization (24–74%) with type strains of the neutrophilic and alkaliphilic Methylophaga species, the isolate was identified as Methylophaga muralis Bur 1 (VKM B-3046 = DSM 103617). The genome of M. muralis Bur 1 contained 2585 protein-encoding genes; 634 proteins with unidentified functions were predicted. Three rRNAs (5S, 16S, and 23S) and 38 tRNAs were identified. Apart from the mxaFJGIRSACKLDEH classical cluster of methanol oxidation genes, the xoxF gene was found. Methylamine was oxidized to formaldehyde by methylamine dehydrogenase and via the N-methylglutamate pathway. Orthologs of type III glutamine synthetases were revealed in the genome. The operons of ectoine and sucrose biosynthesis, ectRABC-ask and sps-spp-fruK-ams, were found. The genomes of M. muralis Bur 1 and M. lonarensis MPL^T, unlike that of M. nitratireducenticrescens JAM1^T, were found to contain the genes encoding the proteins of bicarbonate transport.     

Tepidimonas taiwanensis sp. nov., a novel alkaline-protease-producing bacterium isolated from a hot spring

The bacterial strain designated I1-1^T was isolated from a hot spring located in the Pingtung area, southern Taiwan. Cells of this organism were Gram reaction negative rods, motile by a single polar flagellum. Optimum conditions for growth were 55°C and pH 7. Strain I1-1^T grew well in lower nutrient media such as 5–10% Luria–Bertani broth, and its extracellular products expressed alkaline protease activity. The 16S rRNA gene sequence analysis indicates that strain I1-1^T is a member of -Proteobacteria. On the basis of a phylogenetic analysis of 16S rDNA sequences, DNA–DNA similarity data, whole-cell protein analysis, physiological and biochemical characteristics, as well as fatty acid compositions, the organism belonged to the genus Tepidimonas and represented a novel species within this genus. The predominant cellular fatty acids of strain I1-1^T were 16:0 (about 41%), 18:1 7c (about 13%), and summed feature 3 [16:1 7c or 15:0 iso 2OH or both (about 26%)]. Its DNA base ratio was 68.1 mol%. We propose to classify strain I1-1^T (=BCRC 17406^T=LMG 22826^T) as Tepidimonas taiwanensis sp. nov.     

<Emphasis Type="Italic">Spirosoma pomorum</Emphasis> sp. nov., isolated from apple orchard soil

A Gram-negative, motile, rod-shaped, aerobic bacterial strain, designated S7-2-11^T, was isolated from apple orchard soil from Gyeongsangnam-do Province, Republic of Korea, and was characterized taxonomically using a polyphasic approach. 16S rRNA gene sequence analysis indicated that strain S7-2-11^T belongs to the family Cytophagaceae in phylum Bacteroidetes, and is closely related to Spirosoma luteolum 16F6E^T (94.2% identity), Spirosoma knui 15J8-12^T (92.7%), and Spirosoma linguale DSM 74^T (91.0%). The G + C content of the genomic DNA of strain S7-2-11^T was 49.8 mol%. Strain S7-2-11^T contained summed feature 3 (C_16:1 ω7c/C_16:1 ω6c; 35.1%), C_16:1 ω5c (22.4%), C_15:0 iso (13.9%), and C_17:0 iso 3-OH (10.6%) as major cellular fatty acids, and MK-7 as the predominant respiratory quinone. The main polar lipids were phosphatidylethanolamine, an unidentified aminophospholipid, and two unidentified polar lipids. Phenotypic and chemotaxonomic data supported the affiliation of strain S7-2-11^T with the genus Spirosoma. The results of physiological and biochemical tests showed the genotypic and phenotypic differentiation of the isolate from recognized Spirosoma species. On the basis of its phenotypic properties, genotypic distinctiveness, and chemotaxonomic features, strain S7-2-11^T represents a novel species of the genus Spirosoma, for which the name Spirosoma pomorum sp. nov. is proposed. The type strain is S7-2-11^T (= KCTC 52726^T = JCM 32130^T).     

Role of trehalose synthesis pathways in salt tolerance mechanism of <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> f. sp. <Emphasis Type="Italic">denitrificans</Emphasis> IL106

Two strains of trichloroethylene (TCE)-degrading bacteria were isolated from soils at polluted and unpolluted sites. The isolates, strains TE26^T and K6, showed co-substrate-independent TCE-degrading activity. TCE degradation was accelerated by preincubation with tetrachloroethylene, cis-dichloroethylene (DCE) and 1,1-DCE. TCE-degrading activities of strains TE26^T and K6 were 0.23, 0.24 mol min^–1 g^–1 dry cells, respectively. 16S rDNA sequences of strains TE26^T and K6 were almost identical (99.7% similarity), and most closely related to Ralstonia basilensis (ATCC17697^T) (98.5% similarity). From the results of DNA–DNA hybridizations, strain TE26^T was genetically coherent to strain K6 (94 and 88% hybridization), and exhibited lower relatedness to R. basilensis (DSM11853^T) (44% and 15%). In addition, because of the differences in chemotaxonomic properties, strain TE26^T and strain K6 appear to be distinct from all established species of the Ralstonia group. Based on these results and the proposal of transferring R. basilensis and related species to Wautersia gen. nov., we propose that these strains should be assigned to the genus Wautersia as Wautersia numadzuensis sp. nov.     

<Emphasis Type="Italic">Scopulibacillus darangshiensis</Emphasis> gen. nov., sp. nov., isolated from rock

A novel, Gram-positive bacterium, designated DLS-06^T, was isolated from scoria (volcanic ash) under rock on the peak of small mountain (300 m above the sea level; known as Darangshi Oreum) in Jeju, Republic of Korea. The cells of the isolate were aerobic, oxidase-negative, catalase-positive, endospore-forming, non-motile rods. The organism grew at 25∼30°C and initial pH 6.1∼9.1. A neighbour-joining tree based on 16S rRNA gene sequences showed that the organism was related to members of the family “Sporolactobacillaceae” and related taxa. The phylogenetic neighbours were Pullulanibacillus naganoensis (95.2% 16S rRNA gene sequence similarity), Tuberibacillus calidus (95.0%) and Sporolactobacillus (91.8∼94.2%). Levels of 16S rRNA gene sequence similarity of the isolate to representatives of other genera were in the range of 87.2∼93.7%. The organism contained meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The predominant menaquinone was MK-7. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, an unknown ninhydrin-positive phospholipid, three unknown phospholipids and an unknown lipid. The major fatty acids were anteiso-C_15:0 and anteiso-C_17:0. The G+C content of the DNA was 50.8 mol%. On the basis of the phenotypic and phylogenetic data presented in this study, this organism represents a novel genus and species in the order Bacillales, for which the name Scopulibacillus darangshiensis gen. nov., sp. nov. is proposed. The type strain is DLS-06^T (=DSM 19377^T =KCTC 13161^T).     

<Emphasis Type="Italic">Desulfurispirillum alkaliphilum</Emphasis> gen. nov. sp. nov., a novel obligately anaerobic sulfur- and dissimilatory nitrate-reducing bacterium from a full-scale sulfide-removing bioreactor

Strain SR 1^T was isolated under anaerobic conditions using elemental sulfur as electron acceptor and acetate as carbon and energy source from the Thiopaq bioreactor in Eerbeek (The Netherlands), which is removing H₂S from biogas by oxidation to elemental sulfur under oxygen-limiting and moderately haloalkaline conditions. The bacterium is obligately anaerobic, using elemental sulfur, nitrate and fumarate as electron acceptors. Elemental sulfur is reduced to sulfide through intermediate polysulfide, while nitrate is dissimilatory reduced to ammonium. Furthermore, in the presence of nitrate, strain SR 1^T was able to oxidize limited amounts of sulfide to elemental sulfur during anaerobic growth with acetate. The new isolate is mesophilic and belongs to moderate haloalkaliphiles, with a pH range for growth (on acetate and nitrate) from 7.5 to 10.25 (optimum 9.0), and a salt range from 0.1 to 2.5 M Na⁺ (optimum 0.4 M). According to phylogenetic analysis, SR 1^T is a member of a deep bacterial lineage, distantly related to Chrysiogenes arsenatis (Macy et al. 1996). On the basis of the phenotypic and genetic data, the novel isolate is placed into a new genus and species, Desulfurispirillum alkaliphilum (type strain SR^T = DSM 18275 = UNIQEM U250). Nucleotide sequence accession number: the GenBank/EMBL accession number of the 16S rRNA gene sequence of strain SR 1^T is DQ666683.     

The new alkaliphilic bacteriochlorophyll <Emphasis Type="Italic">a</Emphasis>-containing bacterium <Emphasis Type="Italic">Roseinatronobacter monicus</Emphasis> sp. nov. from the hypersaline Soda Mono Lake (California,United States)

Two strains of pink-colored aerobic bacteriochlorophyll a-containing bacteria were isolated from aerobic (strain ROS 10) and anaerobic (strain ROS 35) zones of the water column of Mono Lake (California, United States). Cells of the bacteria were nonmotile oval gram-negative rods multiplying by binary fission by means of a constriction. No intracellular membranes were detected. Polyphosphates and poly-β-hydroxybutyric acid were the storage compounds. Pigments were represented by bacteriochlorophyll a and carotenoids of the spheroidene series. The strains were obligately aerobic, mesophilic (temperature optimum of 25–30°C), alkaliphilic (pH optimum of 8.5–9.5), and moderately halophilic (optimal NaCl concentration of 40 g/l). They were obligately heterotrophic and grew aerobically in the dark and in the light. Respiration was inhibited by light at wavelengths corresponding to the absorption of the cellular pigments. The substrate utilization spectra were strain-specific. In the course of organotrophic growth, the bacteria could oxidize thiosulfate to sulfate; sulfide and polysulfide could also be oxidized. The DNA G+C content was 59.4 mol % in strain ROS 10 and 59 mol % in strain ROS 35. In their phenotypic properties, the new strains were close but not identical to the alkaliphilic bacterium Roseinatronobacter thiooxidans. The distinctions in the nucleotide sequences of the 16S rRNA genes (2%) and low DNA-DNA hybridization level with Rna. thiooxidans (22–25%) allow the new strains to be assigned to a new species of the genus Roseinatronobacter, Roseinatronobacter monicus sp. nov. with the type strain ROS 35^T (=UNIQEM U-251^T = VKM B-2404^T).     

<Emphasis Type="Italic">Myceligenerans salitolerans</Emphasis> sp. nov., a halotolerant actinomycete isolated from a salt lake in Xinjiang,China

A halotolerant actinomycete strain, designated XHU 5031^T, was isolated from a salt lake in Xinjiang Province, northwest China. Its taxonomic status was determined using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the organism was most closely related to Myceligenerans xiligouense DSM 15700^T (98.4 %), Myceligenerans halotolerans XJEEM 11063^T (98.0 %) and Myceligenerans crystallogenes DSM 17134^T (97.5 %). However, it had relatively low values for DNA–DNA relatedness with the above strains (46.2, 39.4 and 36.5 %, respectively). The peptidoglycan type was A4α. This organism contained glucose, mannose and galactose as the major whole cell sugars. The predominant menaquinone was MK-9(H₄). The major fatty acids were iso-C_15:0, anteiso-C_15:0 and iso-C_16:0. The polar lipids detected were diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylinositol (PI), one unknown phospholipid (PL) and two unknown glycolipids (GL). The G+C content of genomic DNA was 71.2 mol %. Phenotypic data clearly distinguished the isolate from its closest relatives. The combined genotypic and phenotypic data indicated that the isolate XHU 5031^T represented a novel species of the genus Myceligenerans. The proposed name for this organism is Myceligenerans salitolerans sp. nov., with type strain XHU 5031^T (=KCTC 29128^T = CCTCC AB 2012908^T).     

<Emphasis Type="Italic">Methylovorus menthalis</Emphasis>, a novel species of aerobic obligate methylobacteria associated with plants

A bacterial strain (MM) utilizing methanol as the only carbon and energy source was isolated from corn mint rhizoplane. The cells of the strain were gram-negative colorless motile rods. Spores and prosthecae were not formed, reproduced by binary fission, and did not require vitamins and growth factors. The organism was strictly aerobic, urease-, oxidase-, and catalase-positive. Used the KDPG variant of the ribulose monophosphate pathway. Possessed NAD⁺ dependent 6-phosphogluconate dehydrogenase activity and enzymes of the glutamate cycle. The activities of α-ketoglutarate dehydrogenase and of the glyoxylate bypass enzymes (isocitrate lyase and malate synthase) were absent. Palmitic (C_16:0) and palmitoleic (C_16:1) acids were predominant in the cell fatty-acid composition. The dominant phospholipids were phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylcholine. The dominant ubiquinone was Q₈. The strain formed indole from tryptophan. The DNA G + C content was 54.5 mol % (T _m). According to the data of the 16S rRNA gene sequencing, strain MM showed high similarity (98–99%) to Methylovorus glucosotrophus VKM B-1745^T and Methylovorus mays VKM B-2221^T, but the level of DNA-DNA homology with these cultures was only 40 and 58%, respectively. The strain was classified as a new species, Methylovorus menthalis sp. nov. (VKM B-2663^T).