Thermoleophilum album gen. nov. and sp. nov., a bacterium obligate for thermophily and n-alkane substrates

Several bacterial strains that are obligate for both thermophily and hydrocarbon utilization have been isolated from a number of thermal and non-thermal environments. Mud and water samples obtained from geographic sites across the United States were subjected to enrichment procedures at 60° C with n-heptadecane as sole growth substrate. Organisms forming very small white colonies on agar surfaces were often evident on primary enrichment. These bacteria were Gram negative, aerobic, small, and rodshaped. They lacked pigmentation, motility, and the ability to form endospores. Growth occurred in the temperature range from 45° C to 70° C with the optimum around 60° C and at a pH near neutrality. Only n-alkanes from 13 to 20 carbons in length were utilized by these organisms as growth substrate. The mol% guanine plus cytosine values for these strains were between 68 and 70%. The physiological and morphological characteristics of these organisms are distinctly different from any previously described thermophilic microbes. It is proposed that they be placed in a new genus, Thermoleophilum gen. nov. with the type species being Thermoleophilum album gen. nov., sp. nov. The type strain in ATCC 35263.Paper number 8953 of the Journal Series of the North Carolina Agricultural Research Service Raleigh, NC 27695, USA     

Desulfurella acetivorans gen. nov. and sp. nov. —a new thermophilic sulfur-reducing eubacterium

A new type of thermophilic cyanobacterial mat, rich in elemental sulfur and containing large numbers of sulfur-reducing bacteria able to utilize different growth substrates at 55° C, was found in the Uzon caldere (Kamchatka). One of the largest groups among these organisms were acetate-oxidizing sulfur-reducing bacteria, numbering 10⁶ cells · cm^–3 of mat. The pure culture of a sulfur-reducing eubacterium growing on acetate was isolated. Cells of the new isolate are Gram-negative short rods, often in pairs, motile, with a single polar flagellum. The optimal temperature for growth is 52 to 57° C, with no growth observed at 42 or 73° C. The pH optimum is 6.8 to 7.0. The new isolate is demonstrated to be a true dissimilatory sulfur reducer: it is an obligate anaerobe, it is unable to ferment organic substrates and it can use no electron acceptors other than elemental sulfur. Acetate is the only energy and carbon source, and H₂S and CO₂ are growth products. No cytochromes were detected. The G+C content of DNA is rather low, only 31.4 mol%. Thus, morphological and physiological features of the new isolate are quite close to those of Desulfuromonas. But on the grounds of a significant difference in the G+C content of DNA, the absence of cytochromes and because of its thermophilic nature, a new genus Desulfurella is proposed with the type species Desulfurella acetivorans.     

Fervidobacterium nodosum gen. nov. and spec. nov., a new chemoorganotrophic,caldoactive, anaerobic bacterium

A new species of extremely thermophilic, glycolytic anaerobic bacterium, Fervidobacterium nodosum isolated from a New Zealand hot spring, is described. Fervidobacterium nodosum strains were Gram-negative, motile, non-sporulating obligately anaerobic rods that existed singly, in pairs or in chains. Electron micrographs of thin sections revealed a two-layered cell wall structure. The outer layer of the cell wall produced spheroids, which was a typical feature of this organism. The optimum temperature for growth was 65 to 70° C, the maximum 80° C and the minimum greater than 40° C. Growth occurred between a pH of 6.0 and 8.0 with the optimum being 7.0 to 7.5. The doubling time of Fervidobacterium nodosum at optimal temperature and pH was 105 minutes. The DNA base composition was 33.7% guanine plus cytosine as determined by thermal denaturation. A wide range of carbohydrates including glucose, sucrose, starch and lactose could be utilized by the organism. Lactate, acetate, hydrogen, and carbon dioxide were the major end products of glucose fermentation with lesser amounts of ethanol being formed. Growth was inhibited by tetracycline, penicillin and chloramphenicol indicating that the organism was a eubacterium.     

Acidibacter ferrireducens gen. nov., sp. nov.: an acidophilic ferric iron-reducing gammaproteobacterium

An acidophilic gammaproteobacterium, isolated from a pit lake at an abandoned metal mine in south-west Spain, was shown to be distantly related to all characterized prokaryotes, and to be the first representative of a novel genus and species. Isolate MCF85 is a Gram-negative, non-motile, rod-shaped mesophilic bacterium with a temperature growth optimum of 32–35 °C (range 8–45 °C). It was categorized as a moderate acidophile, growing optimally at pH 3.5–4.0 and between pH 2.5 and 4.5. Under optimum conditions its culture doubling time was around 75 min. Only organic electron donors were used by MCF85, and the isolate was confirmed to be an obligate heterotroph. It grew on a limited range of sugars (hexoses and disaccharides, though not pentoses) and some other small molecular weight organic compounds, and growth was partially or completely inhibited by small concentrations of some aliphatic acids. The acidophile grew in the presence of >100 mM ferrous iron or aluminium, but was more sensitive to some other metals, such as copper. It was also much more tolerant of arsenic (V) than arsenic (III). Isolate MCF85 catalysed the reductive dissolution of the ferric iron mineral schwertmannite when incubated under micro-aerobic or anaerobic conditions, causing the culture media pH to increase. There was no evidence, however, that the acidophile could grow by ferric iron respiration under strictly anoxic conditions. Isolate MCF85 is the designated type strain of the novel species Acidibacter ferrireducens (=DSM 27237^T = NCCB 100460^T).     

Thermobacteroides acetoethylicus gen. nov. and spec. nov., a new chemoorganotrophic,anaerobic, thermophilic bacterium

The taxonomic and metabolic characteristics of a new caldoactive bacterium, Thermobacteroides acetoethylicus, that is prevalent in volcanic features where organic matter is vigorously being decomposed is described. T. acetoethylicus is a nonsporing, obligately anaerobic rod that stains gram-negative and exists singly or in pairs. Electron micrographs revealed peritrichous flagellation and a distinctive outer wall envelope structure without an outer wall membrane layer. The temperature range for growth was >40°C and <80°C; the pH range was between 5.5 and 8.5. The DNA base composition was 31±1 mol% guanosine plus cytosine. Fermentable carbohydrates included glucose, mannose, cellobiose, lactose, maltose, sucrose and starch. Growth on glucose in complex medium was associated with a 30 min doubling time; and ethanol, acetate, H₂/CO₂, butyrate and isobutyrate accounted for a balanced fermentation. Lactic acid was not formed. Growth was inhibited by O₂, 2% NaCl, penicillin, streptomycin, vankomycin and neomycin, but not by chloramphenicol or hydrogen (2 atm). Glucose was metabolized via the Embden-Meyerhoff Pathway. A molar growth yield of 18.3 g cell per mol glucose and an ATP yield of 8 g cell per mol ATP produced was obtained. These results support the absence of detectable cytochromes and suggest that energy conservation is via substrate level phosphorylation alone.Abbreviations DNA deoxyribonucleic acid - ATP adenosine triphosphate - LPBB low phosphate buffered basal - TYEG tryptone yeast extract glucose - O.D. optical density - Y _ATP molar ATP yield - G+C guanosine plus cytosine     

Rhodospirillum salexigens,spec. nov., an obligatory halophilic phototrophic bacterium

The name Rhodospirillum salexigens, spec. nov., was proposed for the strain WS 68, isolated by W. R. Sistrom at the Oregon coast from salt water. The spiral-shaped phototrophically or chemotrophically growing, salt demanding bacterium contains intracytoplasmic membranes arranged parallel to the cytoplasmic membrane. Bacteriochlorophyll a and spirilloxanthin are the major pigments. Growth occurs between 20 and 45°C at a neutral pH. The DNA base composition was 64 mol % guanine plus cytosine. The cell wall contains peptidoglycan and proteins but no glycolipids.     

Thermoanaerobium lactoethylicum spec. nov. a new anaerobic bacterium from a hot spring of Kamchatka

A new anaerobic thermophilic Gram-positive, nonsporeforming bacterium strain ZE-1 was isolated from a hot spring of Kamchatka (USSR). The cells are rod-shaped, (0.5–0.8 · 2.0–20 m), non-motile. The bacterium can grow between 42 and 75°C; the optimal temperature is 65°C. The growth is possible between pH values 5.0 and 8.5; optimal pH is 7.0. The cultures grow on the media containing peptone, yeast extract, or casein hydrolysate as nitrogen sources in the presence of glucose or some other sugars, mannitol or starch. The main fermentation products of glucose are ethanol, acetate, lactate, H₂, CO₂; byproducts are propionic, butyric and isovaleric acids. Glucose is metabolized via Embden-Meyerhoff-Parnas pathway. Molecular hydrogen does not inhibit growth. The bacterium does not reduce aceton to isopropanol, but is able to form H₂S from elemental sulfur. The bacterium contains a soluble hydrogenase. This enzyme catalyzes both evolution and uptake of H₂ and is active in the presence of methyl viologen. The DNA-base composition is 34.6 mol%; the genome size 2.08x10⁹ D. The name proposed for the isolated bacterium strain ZE-1 is Thermoanaerobium lactoethylicum spec. nov.     

Heliobacterium sulfidophilum sp. nov. andHeliobacterium undosum sp. nov.: Sulfideoxidizing heliobacteria from thermal sulfidic springs

Two new species of heliobacteria isolated from cyanobacterial mats of two alkaline sulfidic hot springs are formally described. Strains BR4 and BG29 are assigned to anoxygenic phototrophic bacteria of the familyHeliobacteriaceae, since they possess the unique properties of this taxon: strict anaerobiosis, formation of bacteriochlorophyllg, the lack of extensive intracytoplasmic membranes and chlorosomes, an unusual cell wall structure, and phylogenetic relatedness to the low G+C gram-positive eubacteria. Based on the 16S rDNA sequence similarity, strains BR4 and BG29 are assigned to the genusHeliobacterium and described as two new species of this genus:Heliobacterium sulfidophilum sp. nov. andHeliobacterium undosum sp. nov. The G+C content of the DNA is 51.3 mol % inHbt. sulfidophilum and 57.2-57.7 mol % inHbt. undosum. The cells ofHbt. sulfidophilum are rods, and the cells ofHbt. undosum are slightly twisted spirilla or short rods. Both new bacteria are motile by peritrichous flagella.Hbt. sulfidophilum produces endospores. The new bacteria are strict anaerobes growing photoheterotrophically on a limited range of organic compounds. In the dark, they can switch from photosynthesis to the slow fermentation of pyruvate. Biotin is required as a growth factor. Both species are highly tolerant to sulfide (up to 2 mM at pH 7.5) and oxidize it photoheterotrophically to elemental sulfur; photoautotrophic growth was not observed. The temperature optimal for growth ofHbt. sulfidophilum andHbt undosum is 30–35‡C, and the optimal pH is 7–8.     

Desulfurella multipotens sp. nov., a new sulfur-respiring thermophilic eubacterium from Raoul Island (Kermadec archipelago,New Zealand)

A new moderately thermophilic sulfur-reducing eubacterium was isolated from bottom deposits of Green Lake (Raoul Island, Kermadec archipelago, New Zealand). Cells are short rods, 1.5–1.8 by 0.5–0.7 m, single or in pairs, motile with one polar flagellum, gram-negative with S-layer of subunit structure. Growth occurred between 42 and 77°C with the optimum at 58–60°C and at pH from 6.0 to 7.2 with the optimum at 6.4–6.8. The bacterium was obligately anaerobic and obligately sulfur-respiring, and capable of lithoautotrophic growth on a mineral medium with S° and H₂/CO₂ gas phase. In addition to molecular hydrogen, a wide range of substrates can be utilized as energy source in the presence of elemental sulfur: pyruvate, acetate, butyrate, pentadecanate, palmitate, stearate. Products are CO₂ and H₂S. The G+C content of DNA is 33.5 mol%. DNA-DNA homology with the type species of the genus Desulfurella — Desulfurella acetivorans — is 69±2%. A new species, Desulfurella multipotens sp. nov., with the type strain RH-8 is described.     

Thermosediminibacter oceani gen. nov., sp. nov. and Thermosediminibacter litoriperuensis sp. nov., new anaerobic thermophilic bacteria isolated from Peru Margin

A new group of anaerobic thermophilic bacteria was isolated from enrichment cultures obtained from deep sea sediments of Peru Margin collected during Leg 201 of the Ocean Drilling Program. A total of ten isolates were obtained from cores of 1–2 m below seafloor (mbsf) incubated at 60°C: three isolates came from the sediment 426 m below sea level with a surface temperature of 9°C (Site 1227), one from 252 m below sea level with a temperature of 12°C (Site 1228), and six isolates under sulfate-reducing condition from the lower slope of the Peru Trench (Site 1230). Strain JW/IW-1228P from the Site 1228 and strain JW/YJL-1230-7/2 from the Site 1230 were chosen as representatives of the two identified clades. Based on the 16S rDNA sequence analysis, these isolates represent a novel group with Thermovenabulum and Caldanaerobacter as their closest relatives. The temperature range for growth was 52–76°C with an optimum at around 68°C for JW/IW-1228P and 43–76°C with an optimum at around 64°C for JW/YJL-1230-7/2. The pH^25C range for growth was from 6.3 to 9.3 with an optimum at 7.5 for JW/IW-1228P and from 5 to 9.5 with an optimum at 7.9–8.4 for JW/YJL-1230-7/2. The salinity range for growth was from 0% to 6% (w/v) for JW/IW-1228P and from 0% to 4.5% (w/v) for JW/YJL-1230-7/2. The G+C content of the DNA was 50 mol% for both JW/IW-1228P and JW/YJL-1230-7/2. DNA–DNA hybridization yielded 52% similarity between the two strains. According to 16S rRNA gene sequence analysis, the isolates are located within the family, Thermoanaerobacteriaceae. Based on their morphological and physiological properties and phylogenetic analysis, it is proposed that strain JW/IW-1228P^T is placed into a novel taxa, Thermosediminibacter oceani, gen. nov., sp. nov. (DSM 16646^T=ATCC BAA-1034^T), and JW/YJL-1230-7/2^T into Thermosediminibacter litoriperuensis sp. nov. (DSM 16647^T =ATCC BAA-1035^T).An erratum to this article can be found at     

Thermoanaerobium acetigenum spec. nov., a new anaerobic,extremely thermophilic,xylanolytic non-spore-forming bacterium isolated from an Icelandic hot spring

An anaerobic, extremely thermophilic, xylanolytic nonspore-forming bacterium, strain X6B, was isolated from a 70°C Icelandic hot spring sediment. The bacterium was rod-shaped, 3.6–5.9 m long and 0.7 to 1.0 m wide, and cells grew singly, in pairs, and occasionally formed chains. The bacterium was nonmotile with no flagella. Cells from mid-to late exponential gowth-phase cultures stained gram-negative but had a gram-positive like cell wall structure in transmission electron photomicrographs. The bacterium grew between 50°C and 78°C with an optimum temperature at about 65°C to 68°C. Growth occurred between pH 5.2 and 8.5 with an optimum pH close to 7. During growth on beech wood xylan, glucose and d-xylose, the isolate produced CO₂, acetate and H₂ as major fermentation products, and a small amounts of ethanol; lactate was not produced. X6B did not reduce acetone to isopropanol or sulphate or thiosulfate to sulfide. The base composition of X6B's cellular DNA was 35.7 mol% guanine + cytosine. The properties of this strain do not fit any previously described species. The name proposed for the isolated bacterium was Thermoanaerobium acetigenum, spec. nov.     

A new purple sulfur bacterium from saline littoral sediments,Thiorhodovibrio winogradskyi gen. nov. and sp. nov.

Two strains of a new purple sulfur bacterium were isolated in pure culture from the littoral sediment of a saline lake (Mahoney Lake, Canada) and a marine microbial mat from the North Sea island of Mellum, respectively. Single cells were vibrioid-to spirilloid-shaped and motile by means of single polar flagella. Intracellular photosynthetic membranes were of the vesicular type. As photosynthetic pigments, bacteriochlorophyll a and the carotenoids lycopene, rhodopin, anhydrorhodovibrin, rhodovibrin and spirilloxanthin were present.Hydrogen sulfide and elemental sulfur were used under anoxic conditions for phototrophic growth. In addition one strain (06511) used thiosulfate. Carbon dioxide, acetate and pyruvate were utilized by both strains as carbon sources. Depending on the strain propionate, succinate, fumarate, malate, tartrate, malonate, glycerol or peptone may additionally serve as carbon sources in the light. Optimum growth rates were obtained at pH 7.2, 33 °C, 50 mol m^-2 s^-1 intensity of daylight fluorescent tubes and a salinity of 2.2–3.2% NaCl. During growth on sulfide, up to ten small sulfur globules were formed inside the cells. The strains grew microaerophilic in the dark and exhibited high specific respiration rates. No vitamins were required for growth. The DNA base composition was 61.0–62.4 mol% G+C.The newly isolated bacterium belongs to the family chromatiaceae and is described as a member of a new genus and species, Thiorhodovibrio winogradskyi gen. nov. and sp. nov. with the type strain SSP1, DSM No. 6702.     

A thermophilic green sulfur bacterium from New Zealand hot springs,Chlorobium tepidum sp. nov.

Thermophilic green sulfur bacteria of the genus Chlorobium were isolated from certain acidic high sulfide New Zealand hot springs. Cells were Gram-negative nonmotile rods of variable length and contained bacteriochlorophyll c and chlorosomes. Cultures of thermophilic chlorobia grew only under anaerobic, phototrophic conditions, either photoautotrophically or photoheterotrophically. The optimum growth temperature for the strains of thermophilic green sulfur bacteria isolated was 47–48°C with generation times of about 2 h being observed. The upper temperature limit for growth was about 52°C. Thiosulfate was a major electron donor for photoautotrophic growth while sulfide alone was only poorly used. N₂ fixation was observed at 48°C and cell suspensions readily reduced acetylene to ethylene. The G+C content of DNA from strains of thermophilic chlorobia was 56.5–58.2 mol% and the organisms positioned phylogenetically within the green sulfur bacterial branch of the domain Bacteria. The new phototrophs are described as a new species of the genus Chlorobium, Chlorobium tepidum.This paper is dedicated to Professor Norbert Pfennig on the occasion of his 65th birthday     

Thermohydrogenium kirishiense gen. nov. and sp. nov., a new anaerobic thermophilic bacterium

Six strains of a new anaerobic thermophilic non-sporeforming bacterium were isolated in pure culture from industrial yeast biomass. Cells were rod-shaped (0.4–0.8×1.0–11.0 m), non-motile. They stained gram-negative, but outer membrane was not present. The growth occurred between 45–75 °C, the optimal temperature is 65°. Optimal pH value was 7.0–7.4. The bacterium utilized for growth several sugars, starch and yeast extract. The best source of nitrogen was peptone. The main fermentation products of glucose were ethanol, acetate, H₂ and CO₂. As minor products isopropanol, butanol, butyrate and lactate were found. Glucose was metabolized via the Embden-Meyerhoff pathway. Cytochromes and quinones were not found. DNA-base composition was 33.2–34.0 mol%. The DNA-DNA hybridization and 5S rRNA nucleotide sequences showed distantly related of isolated stains to phenotypical similar bacteria. It was proposed to consider the isolated bacterium as Thermohydrogenium kirishiense gen. nov. and sp. nov.     

Rhabdochromatium marinum gen. nom. rev., sp. nov., a purple sulfur bacterium from a salt marsh microbial mat

A new purple sulfur bacterium was isolated in pure culture (strain 8315) from a laminated microbial mat at Great Sippewissett Salt Marsh, Cape Cod, Mass., USA. Single cells were large rods, 10–20 times longer than wide, and predominantly strainght with slightly conical ends. Cells were motile by polarly inserted flagellar tufts. Intracellular photosynthetic membranes were of the vesicular-type. Photosynthetic pigments were bacteriochlorophylla and the carotenoids lycopene, rhodovibrin, anhydrorhodovibrin, and rhodopin. The new bacterium was strictly anaerobic and obligately phototrophic. Hydrogen, hydrogen sulfide, elemental sulfur, and thiosulfate were used as electron donors for photoautotrophic growth. In sulfide-reduced, bicarbonate-containing media, acetate, propionate, and pyruvate were photoassimilated. Growth factors were not required. Optimum growth rates were obtained at pH 7.3, 30°C, a salinity of 1.5–5.0% NaCl, and a light intensity of about 500 lx (tungsten lamp). The DNA base composition of strain 8315 was 60.4 mol% G+C. Comparison of 16S rDNA oligonucleotide catalogue data showed that the new bacterium must be considered a new genus of the Chromatiaceae. The nameRhabdochromatium is revived, and the new speciesRhabdochromatium marinum sp. nov. is described.     

Characterization of a new platelet-forming purple sulfur bacterium,Amoebobacter pedioformis sp. nov.

The dominant purple sulfur bacterium of a reddish-colored waste water pond near Taichung, Taiwan, was isolated in pure culture, strain CML2. Individual cells were nearly spherical, nonmotile, and contained in their peripheral parts was vacuoles that appeared like elongated, curved tubes. Four to sixteen cells formed platelet-like aggregates reminiscent of Thiopedia rosea. The intracellular photosynthetic membrane system of the cells was of vesicular type; the photosynthetic pigments consisted of bacteriochlorophyll a and spirilloxanthin as the major carotenoid. The color of cell suspensions was pink to rosered. Under anaerobic conditions photolithoautotrophic growth occurred with sulfide, elemental sulfur or thiosulfate; sulfur globules were stored as an intermediary oxidation product. In the presence of sulfide, acetate, lactate and pyruvate were photoassimilated; strain CML2 lacked assimilatory sulfate reduction. Fastest photoautotrophic growth (11 h doubling time) was obtained at pH 7.5, 35°C and a light intensity of about 1000 lux (tungsten lamp). Chemolithoautotrophic growth in the dark was possible under reduced oxygen partial pressure with reduced sulfur compounds as respiratory substrates. The DNA base composition of strain CML2 was 65.5 mol% G+C. Strain CML2 is described as type strain of a new species, Amoebobacter pedioformis sp. nov., in the family Chromatiaceae.     

Studies on Aspergilli XVI. Effect of pH,temperature, and carbon and nitrogen interaction

Summary The effect of pH, temperature, and carbon and nitrogen interaction on the growth and sporulation ofAspergillus nidulans (Eidam)Wint.,A. rugulosus Thom &Raper,A. variecolor (Berk. &Br.)Thom &Raper andA. quadrilineatus was studied. All the moulds could grow on a wide range of pH (2.0 to 12.0) but the growth was poor on too acid and too alkaline media. Best growth ofA. rugulosus, A. quadrilineatus, andA. violaceus was seen at pH 6.5 and that ofA. nidulans andA. variecolor at pH 7.0. In general maximum production of perithecia was recorded between pH 6.0 and 8.0.All the above species ofAspergillus under study could grow between a temperature range of 10° C–48° C, but the growth was poor at 10° C and 48° C. The present moulds showed good growth at 20° C, 25°C, and 30° C. At 40° CA. nidulans andA. rugulosus showed moderate growth while the rest of the Aspergilli attained good growth. Temperatures between 20° C–30° C favoured excellent perithecial production.In general, little improvement in growth was noted on media containing good carbon and nitrogen sources. Malic acid was found to be useless when supplied singly. But, poor growth was recorded when supplied in combination with amino acids, amide, and peptone. This was due to the fact that these N sources also supplied carbon for their metabolism.     

Thermotomaculum hydrothermale gen. nov., sp. nov., a novel heterotrophic thermophile within the phylum Acidobacteria from a deep-sea hydrothermal vent chimney in the Southern Okinawa Trough

A novel heterotrophic, thermophilic bacterium, designated strain AC55^T, was isolated from a deep-sea hydrothermal vent chimney at the Hatoma Knoll in the Okinawa Trough, Japan. Cells of strain AC55^T were non-motile, long rods (2.0- to 6.8-μm long and 0.3- to 0.6-μm wide). The strain was an obligatory anaerobic heterotroph capable of fermentative growth on complex proteinaceous substances. Elemental sulfur was reduced to hydrogen sulfide but did not stimulate growth. Growth was observed between 37 and 60°C (optimum 55°C), pH 5.5 and 8.5 (optimum pH 6.6), and in the presence of 1.5–4.5% (w/v) NaCl (optimum 2.5%, w/v). Menaquinone-7 and -8 were the major respiratory quinones. The G + C content of the genomic DNA from strain AC55^T was 51.6 mol%. The 16S rRNA gene sequence analysis revealed that strain AC55^T was the first cultivated representative of Acidobacteria subdivision 10. Based on the physiological and phylogenetic features of the novel isolate, the genus name Thermotomaculum gen. nov. is proposed, with Thermotomaculum hydrothermale sp. nov. as the type species. The type strain is AC55^T (=JCM 17643^T = DSM 24660^T = NBRC 107904^T).     

Ectothiorhodosinus mongolicum gen. nov., sp. nov., a New Purple Bacterium from a Soda Lake in Mongolia

A new nonmotile purple sulfur bacterium (strain M9) was isolated from the steppe soda lake Lake Dzun Uldziin Nur (pH 9.4; mineralization, 3.3%) situated in southeastern Mongolia. Individual cells appear as vibrios 0.3–0.5 × 0.7–1 m in size. The dividing cells often do not separate from each other, forming an almost closed ring. The internal photosynthetic membranes are represented by concentric lamellae lining the cell wall. Photosynthetic pigments are bacteriochlorophyll a and carotenoids of the spirilloxanthin series. The main carotenoid (>96%) is spirilloxanthin. Two typical light-harvesting complexes (LH1 and LH2) are present in the membranes in a 1 : 1 ratio. The bacterium is an anaerobe and facultative photoorganoheterotroph. Photolithoautotrophic growth on sulfide is scarce. Thiosulfate is utilized as an electron donor only in the presence of organic matter. Globules of elemental sulfur are formed as an intermediary product of sulfide and thiosulfate oxidation and are deposited outside the cells. The end product of oxidation is sulfate. In the presence of sulfide and carbonates, acetate, lactate, malate, pyruvate, propionate, succinate, and fumarate are used as additional sources of carbon in anoxygenic photosynthesis. Vitamins are not required. The bacterium is an alkaliphile, the pH optimum is at 8.3–9.1, the pH range is 7.6–10.1. The optimum NaCl concentration in the medium is 1 to 7%; the range is 0.5 to 0.9%. The optimum carbonate content in the medium is 2%; the range is 1 to 10%. The best growth occurs at 30–35°C. The DNA G+C content is 57.5 mol %. According to the results of analysis of the 16S rRNA gene sequences, the new isolate M9 belongs to the phylogenetic cluster containing representatives of the family Ectothiorhodospiraceae within the class Gammaproteobacteria. In this class, the new isolate forms a new branch, which occupies an intermediate position between the representatives of the genera Ectothiorhodospira and Thiorhodospira. Based on the phenotypic and genetic characteristics, the new purple sulfur bacterium was assigned to a new species of a new genus of the family Ectothiorhodospiraceae, Ectothiorhodosinus mongolicum gen. nov., sp. nov.     

Characterization of Aquamicrobium defluvii gen. nov. sp. nov., a thiophene-2-carboxylate-metabolizing bacterium from activated sludge

A gram-negative bacterium was isolated from activated sewage sludge with thiophene-2-carboxylate as the sole source of carbon and with nitrate as an electron acceptor. The isolate, strain NKK, was a motile, oxidase- and catalase-positive, rod-like bacterium with a G+C content of 61.7 mol%. Besides nitrate, oxygen could serve as a terminal electron acceptor. Among many carbon sources tested, only a few sugars, fatty acids, and thiophene-2-carboxylate supported growth. Other heterocyclic compounds were not used. The sulfur atom of thiophene-2-carboxylate was oxidized to thiosulfate when cells were grown aerobically, or to elemental sulfur when cells were grown anaerobically with nitrate. Nitrate was reduced to nitrite. Growth on thiophene-2-carboxylate was dependent on the addition of molybdate to the medium. Tungstate, a specific antagonist of molybdate, inhibited growth on thiophene-2-carboxylate at concentrations > 10^–7 M. Three inducible enzymes involved in the metabolism of thiophene-2-carboxylate were detected: an ATP-, CoA-, thiophene-2-carboxylate- and Mg²⁺-dependent thiophene-2-carboxyl-CoA ligase (AMP-forming), a molybdenum-containing thiophene-2-carboxyl-CoA dehydrogenase, and a thiophene-2-carboxyl-CoA thioesterase. The sequence of the 16S rRNA gene suggested a classification of strain NKK within the α-subgroup of the Proteobacteria as a new genus and species, Aquamicrobium defluvii gen. nov. sp. nov. (DSM 11603), closely related to Mesorhizobium sp. and Phyllobacterium sp., but representing a distinct lineage equal in depth to those of the two mentioned genera. Aquamicrobium defluvii can be distinguished from both genera by a distinct spectrum of substrates, the maximal growth temperature, and a different salt tolerance. Received: 8 September 1997 / Accepted: 14 November 1997