- Volume 35, Issue 4, 1985

We describe two species of the new genus Glycomyces, Glycomyces harbinensis sp. nov. and Glycomyces rutgersensis sp. nov. Members of this genus are aerobic, produce nonfragmenting vegetative hyphae, and form chains of conidia on aerial sporophores. The cell walls are type II (meso-diaminopimelic acid and glycine are present), and the whole-cell sugar patterns are type D (xylose and arabinose are present). The phospholipid pattern of both species is type P-I (no nitrogenous phospholipids). The guanine-plus-cytosine content of the deoxyribonucleic acid ranges from 71 to 73 mol%. The type strain of type species G. harbinensis is strain NRRL 15337 (= LL-D05139), and the type strain of G. rutgersensis is strain NRRL B-16106 (= LL-I-20).

A new species of the genus Thiomicrospira was isolated from the 21°N deep-sea (2,600-m) hydrothermal vent area of the East Pacific Rise. This organism is an obligate chemolithoautotrophic sulfur oxidizer and differs from the two other species of this genus by its deoxyribonucleic acid base composition and by its growth rate and optimal pH in thiosulfate medium. The new species is named Thiomicrospira crunogena and has been deposited in the American Type Culture Collection as strain ATCC 35932, as well as in the Delft Culture Collection as strain LMD 84.00.

Thermobacteroides proteolyticus sp. nov. was isolated from a methanogenic enrichment culture inoculated from a thermophilic digestor (55°C) that was fermenting tannery wastes and cattle manure. The cells were anaerobic, gram-negative, nonsporeforming, nonmotile rods that were 0.5 μm wide and 1 to 6 μm long. At the end of logarithmic growth, they were pleomorphic, with some filamentous cells. The deoxyribonucleic acid base composition was 45 mol% guanine plus cytosine. The temperature optimum was 63°C (growth range 35, to 75°C); the pH optimum was 7.5 (growth range, pH 5.0 to 8.5). The growth substrates used included yeast extract, peptone, casein, gelatin, and Trypticase peptone. Fructose, glucose, maltose, sucrose, and mannose were weakly used as growth substrates; however, addition of yeast extract and either rumen fluid or Trypticase peptone stimulated utilization of these carbohydrates. Acetate, H2, and CO2 were the major products of growth in medium containing gelatin or glucose. The cells were resistant to kanamycin. The type strain is strain BT (= ATCC 35242).

We developed a method for polyacrylamide gel electrophoresis of whole cells of Actinomyces spp. The major advantage of this method is its ease of operation since it obviates the need for the preparation of bacterial extracts. Whole-cell samples were prepared by incubating washed, packed cells in 6 M urea at 37°C for 24 h. The results of disc gel electrophoresis of these whole-cell samples were compared with the results of electrophoresis of soluble protein extracts of the bacteria run under the same conditions. A large number of bands were obtained with the whole-cell preparations, and these bands were resolved better than bands obtained with the protein extracts. A total of 22 strains of Actinomyces spp. were examined by polyacrylamide gel electrophoresis of urea-treated whole cells. A cluster analysis of the polyacrylamide gel electrophoresis band patterns showed that the strains clustered first according to serotype and then according to species. Overall, two main divisions were identified, one containing Actinomyces bovis, Actinomyces odontolyticus, and Actinomyces israelii and one containing Actinomyces viscosus and Actinomyces naeslundii. This method should be a valuable tool in studying the taxonomy of Actinomyces spp.

We describe a new Thiobacillus species which is a gram-negative, motile, rod-shaped organism having polar flagella. The optimum growth temperature is 43 to 45°C, and the optimum pH range is 6.0 to 7.5. This organism is obligately chemolithotrophic and autotrophic and has ribulose bisphosphate carboxylase activity. It is able to oxidize thiosulfate, trithionate, tetrathionate, hexathionate, heptathionate, sulfur, and sulfide, but is not able to use sulfite, thiocyanate, or dithionate for growth. In batch culture it converts thiosulfate to tetrathionate during or before growth. It has both rhodanese and thiosulfate-oxidizing enzyme activites. It does not grow anaerobically with nitrate or nitrous oxide on either thiosulfate or tetrathionate. The guanine-plus-cytosine content of its deoxyribonucleic acid is 66.6 mol%, and it contains ubiquinone Q-8 in its respiratory chain. The organism is named Thiobacillus tepidarius sp. nov. The type strain is strain DSM 3134, which has been deposited in the Deutsche Sammlung von Mikroorganismen, Göttingen, Federal Republic of Germany.

We describe a new species of nonpigmented, saccharolytic Bacteroides, Bacteroides heparinolyticus, which is frequently isolated from human periodontitis lesions. This species produces an enzyme that hydrolyzes heparin. The major product from glucose is succinic acid. B . heparinolyticus strains have membranes typical of gram-negative cells and external surface structures that stain with ruthenium red and glutaraldehyde-osmium. The guanine-plus-cytosine content of the deoxyribonucleic acid is 47 to 49 mol%. Strains of this species have negligible deoxyribonucleic acid homology with other Bacteroides species described previously. Strain HEP (=ATCC 35895) is the type strain. B . heparinolyticus strains are serologically distinct from other Bacteroides species on the basis of cell agglutination and gel diffusion tests.

The historical and present taxonomic and nomenclatural status of the following taxa were examined: “Vibrio terrigenus”Günther 1894, “Vibrio percolans”Mudd and Warren 1923, “Vibrio alcaligenes” Lehmann and Neumann 1927, “Lophomonas alcaligenes” Galarneault and Leifson 1956, “Comamonas percolans”Davis and Park 1962, “Comamonas terrigena” Hugh 1962, “Pseudomonas terrigena” Hugh 1965, “Vibrio cyclosites” Gray and Thornton 1928, “Vibrio neocistes” Gray and Thornton 1928, “Vibrio cuneatus” Gray and Thornton 1928, and “Comamonas compransoris”Nozhevnikova and Zavarzin 1974. Most of these organisms have been misnamed. Deoxyribonucleic acid-ribosomal ribonucleic acid hybridization experiments localized “C. terrigena” (“V. percolans”) NCIB 8193, “V. neocistes” NCIB 2582, and “V. cyclosites” NCIB 2581 on the Pseudomonas acidovorans ribosomal ribonucleic acid branch in the direct Vicinity of Pseudomonas testosteroni. Deoxyribonucleic acid-deoxyribonucleic acid hybridization experiments, serological studies, gel electropherograms of soluble proteins, and a numerical analysis of phenotypic features supported placement of these strains and clinical isolate CCUG 12940 in a single taxon, for which we propose revival of the name Comamonas terrigena Hugh 1962. The type strain is strain NCIB 8193 (= ATCC 8461 = LMG 1253 = CCUG 15327). C. terrigena is the type species of the genus Comamonas Davis and Park 1962 nom. rev. An extensive phenotypic description of C. terrigena is given, and this species can be differentiated from P . acidovorans and P . testosteroni.”Achromobacter cystinovorum” NCIB 4854 is misnamed and was found to be very closely related to P . acidovorans. “V. alcaligenes” NCTC 9239 was found to be misnamed and is closely related to Pseudomonas diminuta, which is itself inappropriately retained in the genus Pseudomonas. The classification of “V. cuneatus”NCIB 8194 as Pseudomonas fluorescens is confirmed. “C. compransoris” DSM 1231 and “V. cyclosites”ATCC 14635 (the so-called synonymous strain of “V. cyclosites” NCIB 2581) are definitely not members of C . terrigena. These two strains differ considerably from each other, as well as from all of the other strains investigated; their exact taxonomic position is unknown.

The new bacterial species Clostridium pfennigii obtained energy for growth by catabolizing pyruvate to acetate and CO2; CO to acetate and butyrate; vanillin to butyrate, protocatechuic aldehyde, and protocatechuate; ferulate to butyrate, caffeate, and hydrocaffeate; and syringate and 3,4,5-trimethoxybenzoate to butyrate and gallate. This new species did not use any other energy source, such as sugars, amino acids, other organic acids (including formate), methanol, ethanol, or H2-CO2. C. pfennigii is a small, motile, anaerobic, gram-positive, monotrichous rod-shaped organism with a lateral to subterminal flagellum, oval subterminal to terminal spores, and a deoxyribonucleic acid guanine-plus-cytosine content of 38 mol%. It did not liquefy gelatin. Based on the features described above, C. pfennigii may be closely related to Acetobacterium woodii. However, strain V5-2T (T = type strain) used pyruvate but did not use sugars or one-carbon compounds other than CO; it produced acetate and butyrate. The stoichiometry of substrate utilization and the growth yields from different energy sources are discussed.

Two new Pseudomonas species which were isolated from rice paddy and clinical specimens (groups Ve-2 and Ve-1) are described. Strains of Pseudomonas oryzihabitans sp. nov. are yellow-pigmented, oxidase-negative, nonsporeforming, gram-negative, polarly monotrichously flagellated, rod-shaped organisms with deoxyribonucleic acid base compositions ranging from 63.9 to 65.6 mol% guanine plus cytosine, ubiquinone Q-9, major cellular fatty acids consisting of C18:1 acid, C16:0 acid, and C16:1 acid, and 3-hydroxy acids consisting of 3-OH-C10:0 acid and 3-OH-C12:0 acid. Strains of this species were isolated from rice paddy and clinical specimens (group Ve-2). The type strain of this species is strain KS0036 (= L-l = AJ 2197 = IAM 1568 = JCM 2952). Strains of Pseudomonas luteola sp. nov. are yellow-pigmented, oxidase-negative, nonsporeforming, gram-negative, polarly multitrichously flagellated, rod-shaped organisms with deoxyribonucleic acid base compositions ranging from 55.4 to 55.9 mol% guanine plus cytosine, ubiquinone Q-9, major cellular fatty acids consisting of C18:1 acid, C16:0 acid, and C16:1 acid, and hydroxy acids consisting of 3-OH-C10:0 acid and 3-OH-C12:0 acid. Strains of this species were isolated from clinical specimens (group Ve-1). The type strain of this species is strain KS0921 (= G. L. Gilardi 4239 = IAM 13000 = JCM 3352).

Two new species, Fusobacterium alocis and Fusobacterium sulci, are described. They were isolated principally from subgingival areas associated with gingivitis or periodontitis. Strains of these species were nonreactive in the biochemical tests usually used and were differentiated most readily from each other and from other Fusobacterium species by patterns of soluble cellular proteins determined by polyacrylamide gel electrophoresis. The type strains of the species are F. alocis ATCC 35896 and F. sulci ATCC 35585.

Clostridium scindens sp. nov., an obligate anaerobe with desmolytic activity, was isolated from human fecal flora. The desmolase, not associated previously with any specific intestinal microorganism, cleaves the carbon-carbon bond of 17-hydroxylated corticoids at C17-C20, thereby converting them to androstans (C19 steroids). In primary cultures on sheep blood agar plates, C. scindens forms minute, nonhemolytic colonies. The gram-positive rods (0.5 to 0.7 by 1 to 2.5 μm) are slightly curved. The rare oval terminal spores (0.8 to 2.0 μm in diameter) are extremely difficult to demonstrate in Gram-stained smears. More than 40% of the cells are fimbriated. Neither a capsule nor flagella are present. d-Fructose, d-glucose, lactose, d-mannose, d-ribose, and d-xylose are fermented. The major fermentation products are acetic acid, ethanol, and hydrogen. The type strain is ATCC 35704 (Bokkenheuser strain 19).

Nucleic acid hybridization studies together with a chemical cell wall analysis and physiological data indicated that some strains designated as Streptococcus sanguis II, Streptococcus sp. (“S. mitior”-S. sanguis), and “S . viridans” I, II, and IV are closely related to the type strain of S. oralis. These strains contained a directly cross-linked peptidoglycan with lysine as a diamino acid and ribitol and choline as characteristic cell wall constituents. We propose that these strains be classified as members of S. oralis. An emended description of S. oralis is given. S . oralis strains are genetically related to S. pneumoniae strains, and representatives of both species contain ribitol and choline in their cell walls.

A mycoplasma isolated from the cloaca of a tortoise was shown to be serologically distinct from 82 recognized Mycoplasma and Acholeplasma species. Three clones obtained from the isolated mycoplasma were examined in detail and proved indistinguishable from each other. One of these strains, 01008T (NCTC 11701), is designated the type strain of a new species, Mycoplasma testudinis.

The morphological, cultural, and physiological properties of a new species of Bacillus are reviewed. This microorganism, for which we propose the name Bacillus thermoruber, is motile, produces terminal or subterminal oval endospores in swollen sporangia, and shows optimal growth at 45 to 48°C. It is characterized by a high deoxyribonucleic acid guanine-plus-cytosine content (57 ±0.8 mol%) and a red, endocellular, nondiffusible pigment. The type strain is strain BT2, which has been deposited in the collection of the Cattedra di Microbiologia Industriale, Università degli Studi di Milano (Italy) as strain MIM 30.8.38.

Cell preparations of Acholeplasma florum L1T, Acholeplasma axanthum S743T, Acholeplasma granularum BTS-39T, Spiroplasma floricola 23-6T, Mycoplasma gallisepticum S6, and Mycoplasma arginini G230T were examined for 13 cytoplasmic enzyme activities involved in the salvage and interconversion of nucleobases, nucleosides, and 5′-mononucleotides. The unique pyrophosphate-dependent nucleoside kinase activity known only in Acholeplasma laidlawii B-PG9 was found in A. axanthum and A. granularum. All the organisms could be divided into three groups based upon their patterns of purine salvage and interconversion activities. All the tested organisms, A. laidlawii, and apparently Mycoplasma mycoides subsp. mycoides lack the ability to synthesize guanylates from other purine mononucleotides, indicating that these members of the class Mollicutes can only salvage guanylates. The plant epiphytes A. florum and S. floricola had an identical purine enzyme pattern which was different from those of all other members of the Mollicutes studied.

A new mesophilic, cellulolytic microorganism was isolated from a municipal dumping ground. The isolation of this bacterium was performed under anaerobic conditions, but its growth was similar under both anaerobic and aerobic conditions. This gram-positive, nonmotile, coryneform rod, with a high (75.8 mol%) guanine-plus-cytosine content in deoxyribonucleic acid (DNA), can grow on numerous carbon sources. The fermentation products are acetic acid, ethanol, formic acid, succinic acid, CO2, and, occasionally, lactic acid. Nucleic acid hybridization tests between the DNA of the isolate and the DNA of Cellulomonas uda ATCC 21399 showed some partial (37 to 40%) relatedness. This information, in addition to other data, suggests that this organism belongs to the genus Cellulomonas, and it is proposed here as a new species, Cellulomonas fermentans sp. nov. The type strain is M (DSM 3133).

Deoxyribonucleic acid relatedness of five strains of Aquaspirillum serpens and also four other species of this genus was assessed by slot blot hybridization. A high degree of genetic relatedness was evident among the strains within the species A. serpens, but a low degree of relatedness was shown between the species A. serpens, A. sinuosum, A. itersonii, and A. putridiconchylium. Hybridization homology indicated that A. bengal is a subjective synonym for A. serpens and should not have separate species status; therefore, the description of A. serpens is emended accordingly. The strains that grow optimally at higher temperatures (41°C) and produce brown pigments in the presence of some amino acids may be considered varietal and are referred to as A. serpens biovar bengal. This study demonstrates the usefulness of slot blot hybridization for quickly assessing the relatedness of different taxonomic groups of bacteria.

A new genus, Stella, is proposed for a group of flat, six-pronged star-shaped prosthecobacteria found in freshwater, soil, and sewage in widely separated geographical areas. The morphology of the cells provides clear differentiation from all other eubacteria. The cellular morphology and mode of multiplication of these organisms also provide clear separation from other budding and appendaged prosthecobacteria. The cells are gram negative, flat, nonmotile, and asporogenic. Representative organisms in the genus are chemoorganotrophic, growing on low nutrient concentrations and utilizing a variety of amino acids and organic acids. Two distinct species are recognized. Stella humosa (type species) is an avacuolate organism, and the type strain (strain AUCM B-1137) has been deposited in the All Union Collection of Microorganisms, Moscow, USSR, and in the Institute of Microbiology, Academy of Sciences, Moscow, USSR. Stella vacuolata is a gas vacuolate prosthecobacterium, and the type strain is strain 229 (Institute of Microbiology, Academy of Sciences, Moscow, USSR). Organisms in the genus Stella appear to represent the first example of radial symmetry in procaryotic cells.