- Volume 43, Issue 3, 1993
Volume 43, Issue 3, 1993
- Original Papers Relating To Systematic Bacteriology
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Natronobacterium vacuolata sp. nov., a Haloalkaliphilic Archaeon Isolated from Lake Magadi, Kenya
More LessAbstractA novel haloalkaliphilic archaeon was isolated from Lake Magadi, a Kenyan alkaline soda lake. Cells of the organism contain large gas vacuoles in the stationary phase of growth, and colonies produced by these archaea are bright pink in appearance. The major polar lipids of these organisms are C20C20 and C20C25 derivatives of phosphatidylglycerol phosphate and phosphatidylglycerol, and the organisms contain an unidentified phospholipid as a minor component. The G+C content of the DNA is 62.7 mol%. The name Natronobacterium vacuolata sp. nov. is proposed. The type strain is designated NCIMB 13189.
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Mycobacterium brumae sp. nov., a Rapidly Growing, Nonphotochromogenic Mycobacterium
AbstractStrains of a new species of rapidly growing, nonphotochromogenic mycobacteria, Mycobacterium brumae, have been isolated from water, soil, and human sputum samples in Barcelona, Spain. The inclusion of this organism in the genus Mycobacterium is based on its acid-alcohol fastness, its DNA G+C content, its mycolate pattern, and its mycolate pyrolysis esters. A study of 11 strains showed that they form a homogeneous group with an internal phenotypic similarity value of 94.9 ± 3.79%. The results of a comparison with 39 other mycobacterial species and subspecies are also presented. DNA relatedness studies showed that the M. brumae strains studied form a single DNA hybridization group which is less than 30% related to 15 other species of the genus Mycobacterium. Thin-layer chromatographic analysis showed that only α-mycolates are present. Unlike Mycobacterium fallax and Mycobacterium triviale α-mycolates, M. brumae α-mycolates release only 22-carbon atom esters after pyrolysis. Strain CR-270 is the type strain; a culture of this strain has been deposited in the Collection Nationale de Cultures de Microorganismes de l’Institut Pasteur, Paris, France, as strain CIP 103465.
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Fatty Acid and Polar Lipid Composition of the Genus Amycolatopsis: Application of Fast Atom Bombardment-Mass Spectrometry to Structure Analysis of Underivatized Phospholipids
More LessAbstractPhospholipid patterns of 15 representative strains of the genus Amycolatopsis were recorded by twodimensional thin-layer chromatography. The structure analysis of the isolated phospholipids was verified by fast atom bombardment-mass spectroscopy. The positive- and negative-ion spectra of the partially purified phospholipid fractions qualitatively reflect their distinctive composition. All strains contained diphosphatidyl- glycerol, phosphatidylglycerol, and phosphatidylinositol. Two different types of phosphatidylethanolamine and phosphatidylmethylethanolamine were detected, viz., compounds with or without hydroxy fatty acids. These phospholipid patterns underline the integrity of the genus. Fast atom bombardment-mass spectrometry analysis of phospholipid patterns may serve as an aid for differentiation of bacterial species.
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Spiroplasma cantharicola sp. nov., from Cantharid Beetles (Coleoptera: Cantharidae)
AbstractSpiroplasma strain CC-1T, isolated from the gut of the soldier beetle Cantharis carolinus, was serologically distinct from other spiroplasma species, groups, and subgroups. Cells of strain CC-1T were shown by light microscopy to be helical, motile filaments. Electron microscopy showed that the cells were bounded by a single cytoplasmic membrane, with no evidence of a cell wall. The organism was insensitive to penicillin. Strain CC-1T grew well in SM-1, MID, and SP-4 liquid media under aerobic or anaerobic conditions. The strain also grew in 1% serum fraction medium. Optimal growth occurred at 32°C, with a doubling time of 2.6 h, but the strain multiplied at temperatures of 10 to 37°C. Strain CC-1T produced acid from glucose but hydrolyzed neither arginine nor urea. The guanine-plus-cytosine (G+C) content of the DNA was 26 ± 1 mol%. Other uncloned isolates from C. carolinus exhibited similar or identical serological patterns. On the basis of the data presented here, strain CC-1T (= ATCC 43207), previously proposed as the representative strain of subgroup XVI-1, is designated the type strain of a new species, Spiroplasma cantharicola.
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Phylogenetic Analysis of Some Aerococcus-Like Organisms from Clinical Sources: Description of Helcococcus kunzii gen. nov., sp. nov.
More LessAbstract16S rRNA gene sequencing studies were performed on some unusual gram-positive catalase-negative cocci of unknown taxonomic position isolated from human clinical sources. Comparative analysis of the sequence data demonstrated that the clinical isolates represent a hitherto-unknown line of descent within the low-G+C- content gram-positive bacteria. On the basis of the phylogenetic findings and the phenotypic distinctiveness of the organisms, it is proposed that they be classified in a new genus, Helcococcus, as Helcococcus kunzii sp. nov. The type strain of H. kunzii is NCFB 2900.
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Isolation and Characterization of Methanohalophilus portucalensis sp. nov. and DNA Reassociation Study of the Genus Methanohalophilus
More LessAbstractSix strains of coccoid, halophilic methanogens were isolated from various salinaria and natural hypersaline environments. These isolates (strains FDF-1T [T = type strain], FDF-2, SF-2, Ret-1, SD-1, and Cas-1) grew on media containing methanol and mono-, di-, and trimethylamines as catabolic substrates, but not on media containing dimethyl sulfide, methane thiol, H2, formate, or acetate; when cells were provided with H2 in addition to methanol or trimethylamine, they grew on the medium containing a methyl substrate but did not catabolize H2. All of the strains were capable of growth in mineral medium to which trimethylamine was added as a catabolic substrate, although some strains were greatly stimulated by biotin or p-aminobenzoate. DNA reassociation and denaturing electrophoresis of whole-cell proteins indicated that strains FDF-1T, FDF-2, SF-2, and Ret-1, together with previously described strains SF-1, Z-7302, Z-7401, Z-7404, and Z-7405, belong to a new taxon named Methanohalophilus portucalensis sp. nov; FDF-1 (= OCM 59) is the type strain. These strains grew fastest at temperatures near 40°C and, in medium containing 0.5 to 2.5 M NaCl, at pH values near 7. The two new strains excluded from the species on the basis of the results of phylogenetic tests, strains Cas-1 and SD-1, also differed from M. portucalensis in some minor physiological characteristics. Strain Cas-1 was less halophilic (fastest growth occurred in the presence of 0.5 to 1 M NaCl), and strain SD-1 was slightly alkaliphilic (fastest growth occurred at pH 7.8). The DNA reassociation study also showed that Methanohalophilus mahii SLPT exhibited 52% sequence similarity with Methanohalophilus halophilus Z-7982T, supporting the classification of these organisms as separate but closely related species.
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Polyphasic Taxonomy Leading to the Proposal of Moraxella canis sp. nov. for Moraxella catarrhalis-Like, Strains
AbstractThe taxonomic position of a group of 16 Moraxella catarrhalis-like strains, isolated mainly from dogs, was examined by using morphological tests, biochemical tests, serology, ribotyping with oligonucleotide probes, polymerase chain reaction typing of the 16S rRNA gene and the 16S-23S rRNA gene spacer region, polyacrylamide gel electrophoresis of total proteins, fatty acid profiles, moles percent G+C, dot spot and in-solution DNA-DNA hybridizations, and DNA-rRNA hybridizations. It was found that these organisms constitute a distinct cluster within the genus Moraxella. Since they differ genotypically as well as phenotypically from previously described Moraxella species, a new species, Moraxella canis, is proposed to accommodate these isolates. The type strain is LMG 11194 (= N7 = CCUG 8415A).
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Isolation and Characterization of a Moderately Thermophilic Anaerobic Alkaliphile, Clostridium paradoxum sp. nov.
More LessAbstractAlkaliphilic, moderately thermophilic anaerobic bacteria able to grow above pH 10.5 and 55°C were isolated from various sewage plants in the United States. The strains were motile with two to six peritrichous flagella and formed round to slightly oval terminal spores in terminally distended and slightly enlarged cells. Sporulated cells remained motile. The pH range for growth was between 7.0 and 11.1, with an optimum of around 10.1. At pH 10.1 the temperature range for growth was between 30 and 63°C, with an optimum of 56°C. The shortest observed doubling time (glucose) was around 16 min at 56°C and pH 10.1. No dissimilatory sulfate reduction was detected. The organism utilized glucose, fructose, sucrose, maltose, and pyruvate but required yeast extract or tryptone for growth. Optimal NaCl concentrations for growth were between 50 and 200 mM. The guanine-plus-cytosine content was 30.0 ± 0.10 mol%. On the basis of unique properties and 16S rRNA analysis, the strains are placed in a new species, Clostridium paradoxum, referring to the unusual retainment of motility by sporulated cells. Strain JW-YL-7 (DSM 7308) is designated as the type strain.
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Phylogenetic Diversity of Phytopathogenic Mycoplasmalike Organisms
More LessAbstractBy using specific primers, the 16S rRNA genes of Japanese mycoplasmalike organisms (MLOs) were amplified by polymerase chain reactions from MLO-enriched fractions of plants infected with each of six different MLOs. Each of the polymerase chain reaction fragments (length, 1,370 nucleotides) was directly sequenced in both strands by using 17 oligonucleotide primers. A phylogenetic tree constructed by using the sequence data showed that these Japanese MLOs are phylogenetically diverse microorganisms that fall into three groups, group I (onion yellows, tomato yellows, mulberry dwarf, and paulownia witches’ broom MLOs), group II (tsuwabuki witches’ broom MLO), and group III (rice yellow dwarf MLO). A high level of sequence homology (99%) between the Oenothera hookeri MLO and the severe strain of the western aster yellows MLO on the one hand and group I MLOs on the other indicates that the O. hookeri MLO and the severe strain of the western aster yellows MLO belong to group I and suggests that these MLOs, isolated from two geographically separated locations, descended from a very similar ancestor. Although group I contains phylogenetically identical MLOs, the organisms are transmitted by diverse insect vectors. The three MLO groups are more closely related to Acholeplasma laidlawii than to Mycoplasma gallisepticum. Thus, although MLOs are phylogenetically diverse, they are evolutionarily distant from other mollicutes. These data, together with other information (including phylogenetic relationships, vector specificity, plant-pathogenic properties, and habitat in plant phloem sieve tubes), suggest that MLOs could be classified into at least three phylogenetic groups (groups I through III).
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A New Genus of Actinomycetes, Planotetraspora gen. nov.
More LessAbstractA new aerobic mesophilic genus of actinomycetes, Planotetraspora, is described. These microorganisms produce a filamentous growth which is differentiated into a vegetative mycelium and an aerial mycelium; the hyphae are gram positive and not acid fast. Planotetraspora strains are characterized by the formation of long, cylindrical sporangia at the ends of short sporangiophores on aerial hyphae, with each sporangium containing four spores in a single row. The sporangiospores are released from the sporangia when preparations are flooded with water and are motile by means of single polar flagella. The type species of the new genus is Planotetraspora mira (type strain, NA9211028 [= SIIA9201]).
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Molecular Taxonomic Studies of Actinomyces-Like Bacteria Isolated from Purulent Lesions in Pigs and Description of Actinomyces hyovaginalis sp. nov.
More LessAbstractThe 16S rRNA gene sequence of some Actinomyces-like bacteria isolated from purulent lesions in pigs was determined. A comparative analysis of the rRNA sequence data revealed that the bacteria are members of the genus Actinomyces, but are phylogenetically distinct from Actinomyces suis. On the basis of our findings and the results of previous phenotypic studies it is formally proposed that the bacteria from pigs should be designated a new species, Actinomyces hyovaginalis.
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Moraxella lincolnii sp. nov., Isolated from the Human Respiratory Tract, and Reevaluation of the Taxonomic Position of Moraxella osloensis
More LessAbstractA polyphasic taxonomic study was performed to determine the relationships of 10 Moraxella-like strains isolated mainly from the human respiratoiy tract in Sweden. Two of the strains formed a separate subgroup on the basis of both their protein contents and their fatty acid contents. However, the overall protein and fatty acid profiles revealed that all 10 strains were highly related. Representative strains of the two subgroups exhibited high DNA binding values (98%) with each other and had an identical DNA base ratio (44 mol% G+C). DNA-rRNA hybridizations revealed that this taxon can be included in the genus Moraxella, which is only distantly related to phenotypically similar genera, such as the genera Neisseria and Kingella. The results of an extensive phenotypic analysis indicated that the general biochemical profile of the 10 strains conforms with the description of the genus Moraxella given in Bergey’s Manual of Systematic Bacteriology. We therefore consider these organisms members of a new Moraxella species, for which the name Moraxella lincolnii is proposed. Furthermore, we also conclude that Moraxella osloensis belongs, genotypically as well as phenotypically, to the genus Moraxella.
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Serovar Determination and Molecular Taxonomic Correlation in Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum: A Cooperative Study of the International Working Group on Mycobacterial Taxonomy
A cooperative study was conducted by the International Working Group on Mycobacterial Taxonomy to correlate the agglutination serovar designations of Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum strains with the species ascriptions of these organisms according to molecular criteria and cultural properties and to assess the reproducibility of serovar determinations for a set of 63 reference strains of these species. Among the molecular criteria, the level of agreement between results obtained with nucleic acid probes and T-catalase serology results was 94% for strains of M. avium and M. intracellulare. Nucleic acid probes were not available for M. scrofulaceum, but none of the 10 strains ascribed to this species on the basis of catalase serology data reacted with a nucleic acid probe for M. avium or M. intracellulare. Ascription to a species on the basis of mycolic acid high-performance liquid chromatography patterns was in agreement with catalase serology results in 86% of the cases examined. Most strains belonging to serovars 1 through 6 and 8 through 11 were identified by molecular criteria as M. avium, most strains belonging to serovars 7, 12 through 20, 23, and 25 were identified as M. intracellulare, and most strains belonging to serovars 41 through 43 were identified as M. scrofulaceum, in agreement with common current practice. Evidence for assigning serovar 27 to M. scrofulaceum was obtained. However, two strains of a given serovar may, on occasion, be placed in different species. The dominant species assignments for strains belonging to serovars 21, 24, 26, and 28 remain unresolved. Data from laboratories which used panels of sera that corresponded to all of the serovars represented in the study were in agreement with the consensus results in 84% of the instances. Laboratories in which a limited panel of sera was used were far more likely to report results that did not agree with the consensus results.
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Phylogeny of Species in the Family Neisseriaceae Isolated from Human Dental Plaque and Description of Kingella orale sp. nov.
More LessFourteen human periodontal isolates recovered from a purported Eikenella corrodens–selective medium containing 1 μg of clindamycin per ml displayed biochemical traits which differed from those described for E. corrodens. These organisms were gram-negative rods which corroded agar. The isolates were oxidase positive and urease, indole, and esculin negative. They differed from E. corrodens in catalase, nitrate reduction, lysine decarboxylase, and ornithine decarboxylase activities. One isolate, strain UB-294, was presumptively identified as Kingella denitrificans. A second isolate, strain UB-204, differed from E. corrodens by being catalase positive and nitrate reduction negative. Twelve isolates, including strain UB-38T (T = type strain), were phenotypically similar to Kingella kingae except that they did not produce acid from maltose and were not beta-hemolytic. Essentially complete (1,480-base) 16S rRNA sequences were determined for strains UB-38T, UB-204, and UB-294 and the type strains of Neisseria animalis, Neisseria canis, Neisseria denitrificans, Neisseria elongata, Neisseria flavescens, Neisseria macaca, and Neisseria polysaccharea. These sequences were compared with the previously published sequences of six other species belonging to the family Neisseriaceae. On the basis of the results of the comparative sequence analysis, UB-294 was confirmed as a K. denitrificans strain, UB-204 was identified as a member of a new species which may belong in the genus Eikenella, and UB-38T was identified as a member of a new species of the genus Kingella, for which we propose the name Kingella orale. Since strain UB-204 was the only representative of a new species, it was not named. DNA probes for identification of E. corrodens, K. denitrificans, and K. orale based on 16S rRNA sequence information are described.
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DNA Relatedness among Nonpigmented Species of Alteromonas and Synonymy of Alteromonas haloplanktis (ZoBell and Upham 1944) Reichelt and Baumann 1973 and Alteromonas tetraodonis Simidu et al. 1990
More LessThe taxonomic relationships among nonpigmented species of the genus Alteromonas were assessed by DNA-DNA hybridization. The species formed two groups based on DNA homology values and moles precent G+C of DNA: Alteromonas macleodii, the type species, had a moles percent G+C content 4 to 7% higher than those of the other group, and it had less than 5% DNA homology to the second group formed by the remaining species. The remaining species formed seven DNA homology subgroups, of which five corresponded to the type strains of A. atlantica, A. espejiana, A. carrageenovora, A. undina, and A. nigrifaciens. A sixth subgroup included the type strains of A. haloplanktis and A. tetraodonis, which showed hybridizations of more than 80%. The species in the second group were related to each other at the genus level, but a nomenclatural proposal is postponed. The hybridization data showed that the name A. tetraodonis Simidu et al. 1990 should be recognized as a junior subjective synomyn of A. haloplanktis (ZoBell and Upham 1944) Reichelt and Baumann 1973. The seventh subgroup included two as yet unassigned strains which were misidentified as to species.
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Further Studies of the Genus Methylobacterium and Description of Methylobacterium aminovorans sp. nov.
More LessPink-pigmented facultatively methylotrophs belonging to the genus Methylobacterium were examined in this study. Methylobacterium strains are divided into eight species on the basis of phenotypic characteristics. This division is supported by the results of DNA-DNA homology experiments. On the basis of DNA relatedness and other characteristics, a new species, Methylobacterium aminovorans, is proposed. The type strain of M. aminovorans is strain TH-15 (= JCM 8240).
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Arhodomonas aquaeolei gen. nov., sp. nov., an Aerobic, Halophilic Bacterium Isolated from a Subterranean Brine
More LessArhodomonas aquaeolei gen. nov., sp. nov., isolated from a petroleum reservoir production fluid, is described. The single isolate was an obligately halophilic, aerobic, gram-negative, oval rod-shaped bacterium that was actively motile by means of a single polar flagellum. It was catalase and oxidase positive. The isolate had a specific requirement for NaCI; growth occurred at NaCI concentrations between 6 and 20%, and optimal growth occurred in the presence of 15% NaCI. This species metabolized primarily organic acids and required biotin for growth. The name Arhodomonas is proposed for the new genus, which was placed in the gamma subclass of the Proteobacteria on the basis of the results of a 16S rRNA sequence analysis. Although A. aquaeolei is most closely related to purple sulfur bacteria (the genera Ectothiorhodospira and Chromatium), it is not a phototrophic microorganism, which is consistent with its isolation from a subterranean environment. The major components of its cellular fatty acids were C16:0, C18:1, C19:0, C16:1, and C18:0 acids. The DNA base composition of the type strain is 67 mol% G+C. The type and only strain is strain HA-1 (= ATCC 49307).
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Confirmation that Rickettsia helvetica sp. nov. Is a Distinct Species of the Spotted Fever Group of Rickettsiae
More LessWe propose the name Rickettsia helvetica sp. nov. for a rickettsial serotype of unknown pathogenicity isolated in 1979 in Switzerland from Ixodes ricinus ticks and designated the Swiss agent. The growth characteristics and the results of microimmunofluorescence serologic typing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blotting (immunoblotting) with specific mice sera, and a polymerase chain reaction followed by restriction fragment length polymorphism analysis confirmed previously reported preliminary findings which suggested that this rickettsia, to which a name was given provisionally, does represent a new member of the spotted fever group of rickettsiae. The type strain is C3 (Reference Center for Rickettsioses, Marseille, France).
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A Test for Measuring Growth Responses of Mollicutes to Serum and Polyoxyethylene Sorbitan
More LessA test is described that is useful for characterizing mollicutes in terms of the ability to maintain growth in medium containing 15 to 20% fetal bovine serum or in serum-free media with or without 0.04% Tween 80 (polyoxyethylene sorbitan). Representative Acholeplasma species maintained growth in serum-free medium, and about half of the strains tested grew well in Tween 80-supplemented medium. Representative Mycoplasma and Entomoplasma species did not maintain growth in either serum-free medium alone or when Tween 80 was added. Spiroplasma species and group representatives generally failed to sustain growth in serum-free medium with or without Tween 80, but at least four of the spiroplasmas tested maintained growth in serum-free medium. The representative Mesoplasma species grew in serum-free media only when Tween 80 was added, as did Mycoplasma lactucae. Although the test has obvious determinative uses for members of the class Mollicutes, it does not supplant the conventional methodology for assaying the cholesterol requirements of these organisms.
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Ileal Symbiont Intracellularis, an Obligate Intracellular Bacterium of Porcine Intestines Showing a Relationship to Desulfovibrio Species
More LessA new genus and species of obligate intracellular bacteria found in porcine intestines are described. Growth on any bacteriological medium deprived of living cells has not been demonstrated. The organism has been grown intracellularly in cell culture. The 16S rRNA gene sequence data, DNA probe results, and microscopic observations provide evidence that these bacteria differ from those in other described genera and that they belong to the delta subdivision of the class Proteobacteria. We have amplified and sequenced the 16S ribosomal DNA of four preparations of the intracellular bacterium from pigs. For this, intracellular organisms were released and purified from the infected cells without culture techniques. After DNA purification, the polymerase chain reaction with primers complementary to highly conserved eubacterial sequences was used to amplify regions of 16S ribosomal DNA which were subsequently cloned (in some cases) and sequenced directly by standard techniques. The sequences obtained from each preparation were identical and were most similar to that of a sulfate-reducing proteobacterium, Desulfovibrio desulfuricans ATCC 27774 (91% similarity). An oligonucleotide probe complementary to a hypervariable region of the 16S rRNA sequence of the bacterium hybridized with intracellular organisms obtained from porcine intestines. The bacterium is a gram-negative, curved rod with tapered ends. It multiplies intracellularly in the cytoplasm of ileal epithelial cells by septation. The vernacular name Ileal symbiont (IS) intracellularis is proposed for this bacterium. The type strain of IS intracellularis is strain 1482/89 grown in cell culture from a pig affected by proliferative enteropathy. It is deposited in the National Collection of Type Cultures, Colindale, London, as NCTC 12656.
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Mycobacterium celatum sp. nov.
A new slowly growing nonphotochromogenic Mycobacterium species of clinical importance is described. The biochemical characteristics of this organism were similar to those of Mycobacterium xenopi and members of the Mycobacterium avium complex. However, none of the strains reacted with commercially available genetic probes for the M. avium complex. The strains were resistant to most antituberculosis drugs. Multilocus enzyme electrophoresis revealed two original electrophoretic types, which was suggestive of new species. The strains contained α-, keto-, and dicarboxylic mycolates, as determined by thin-layer chromatography. A mycolic acid analysis by high-performance liquid chromatography revealed a chromatographic pattern similar to that of M. xenopi, but distinct from the patterns of previously described Mycobacterium species. Hexadecanoic and tuberculostearic acids were identified as the major cell wall fatty acids by gas-liquid chromatographic analysis; hexacosanoic acid was the major mycolic acid cleavage product, and 2-eicosanol was the major alcohol. Evaluation of the 16S rRNA sequence confirmed the phylogenetic position of the organism among the slowly growing Mycobacterium species. Cultures representing this new species have been deposited in the American Type Culture Collection as strains ATCC 51130 and ATCC 51131T (T = type strain). The name Mycobacterium celatum is proposed.
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Proposal of Two New Species in the Genus Microbacterium: Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov.
More LessThe taxonomic positions of Flavobacterium sp. strain IFO 14592T (= M-73T) (T = type strain), a dextran-α-1,2-debranching enzyme producer, and Microbacterium sp. strain IFO 15204T (= H-5T), an isolate obtained from corn steep liquor, were investigated; on the basis of the results of chemotaxonomic and phenetic studies and DNA-DNA similarity data, we propose that these bacteria should be classified in the genus Microbacterium as Microbacterium dextranolyticum sp. nov. and Microbacterium aurum sp. nov., respectively. The type strain of M. dextranolyticum is strain IFO 14592, and the type strain of M. aurum is strain IFO 15204.
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Proposal of Six New Species in the Genus Aureobacterium and Transfer of Flavobacterium esteraromaticum Omelianski to the Genus Aureobacterium as Aureobacterium esteraromaticum comb. nov.
More LessTwelve strains placed in the genera Flavobacterium, Pseudomonas, and Aureobacterium, including soil isolates, were characterized taxonomically. On the basis of morphological, physiological, and chemotaxonomic data, as well as DNA-DNA hybridization data, we propose that 11 of these strains should be classified in the genus Aureobacterium as new combinations or new species, as follows: Aureobacterium esteraromaticum comb. nov. (type strain, IFO 3751 [= ATCC 8091]), Aureobacterium arabinogalactanolyticum sp. nov. (type strain, IFO 14344), Aureobacterium keratanolyticum sp. nov. (type strain, IFO 13309), Aureobacterium luteolum sp. nov. (type strain, IFO 15074 [= DSM 20143]), Aureobacterium schleiferi sp. nov. (type strain, IFO 15075 [= DSM 20489]), Aureobacterium terrae sp. nov. (type strain, IFO 15300), and Aureobacterium trichothecenolyticum sp. nov. (type strain, IFO 15077 [= JCM 1358]). Whereas the peptidoglycan type of members of this genus is considered to be B2β, the new species A. keratanolyticum was shown to have a new peptidoglycan type, murein variation B2β. An emended description of the genus Aureobacterium is presented.
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Phenotypic and Genomic Analyses of Human Strains Belonging or Related to Bifidobacterium longum, Bifidobacterium infantis, and Bifidobacterium breve
More LessA numerical analysis based on phenotypic characteristics (89 enzymatic tests and 49 carbohydrate acidification tests), in which experimental strips from Biomerieux-API, La Balme les Grottes, France, were used, was performed to characterize 82 new isolates belonging or related to Bifidobacterium longum, Bifidobacterium infantis, and Bifidobacterium breve. A total of 72 strains were isolated from child or adult feces, and the other strains were obtained from human vaginas and bronchi. In this study we also included 38 type and reference strains that were representative of all speices of the genus Bifidobacterium and 6 strains belonging to the genus Lactobacillus. DNA-DNA relationships between B. longum and B. infantis were determined by using 19 strains related to these species, as determined by the numerical analysis. The degree of DNA binding was determined by the S1 nuclease method. The phenotypic study revealed that there were six main clusters, which were subdivided into nine subclusters. Subcluster Va contained the type strains of B. longun and B. infantis. The DNA-DNA relatedness values of some of the new isolates were very similar to the DNA-DNA relatedness values of the type strain of B. longum. On the basis of these data, it was difficult to isolate B. infantis strains and then to define B. infantis as a single species separated from B. longum. Subclusters IVb to IVf comprised reference strains of B. breve. Cluster III and subcluster Ia were not identified.
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Azoarcus gen. nov., Nitrogen-Fixing Proteobacteria Associated with Roots of Kallar Grass (Leptochloa fusca (L.) Kunth), and Description of Two Species, Azoarcus indigens sp. nov. and Azoarcus communis sp. nov.
Among the nitrogen-fixing bacteria associated with roots of Leptochloa fusca (L.) Kunth in saline-sodic soils in the Punjab of Pakistan, we repeatedly found yellow-pigmented, straight to curved, gram-negative rods. To group and identify these organisms, we examined morphological, nutritional, and biochemical features and performed polyacrylamide gel electrophoretic analyses of cellular proteins, gas chromatographic analyses of fatty acids, DNA-rRNA hybridizations, and DNA-DNA hybridizations. Our results showed that 11 isolates formed five groups distinct at the species level, with each group containing one to three members. These bacteria constituted a separate rRNA branch in rRNA superfamily III (corresponding to the beta subclass of the Proteobacteria) at a branching Tm(e) level of 67.7°C [Tm(e) is the temperature at which 50% of a hybrid is denatured under standard conditions]. On this branch, the five groups were located in two clusters with Tm(e) values of 79.4 to 80.4°C and around 71.5°C. We propose a new genus, the genus Azoarcus, for these strains. Azoarcus indigens is the type species and has a growth factor requirement; its type strain is strain VB32 (= LMG 9092). A second named species, Azoarcus communis, includes a strain obtained from French refinery oily sludge, strain LMG 5514. Bacteria of this genus have a strictly aerobic type of metabolism, fix nitrogen microaerobically, and grow well on salts of organic acids but not on carbohydrates. Swedish isolates obtained from human sources (E. Falsen group 15 strains LMG 6115 and LMG 6116), as well as “[Pseudomonas] gasotropha” LMG 7583T, were also located on this rRNA branch at a lower Tm(e) level (70.4 to 71.2°C).
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Mycoplasma corogypsi sp. nov., a New Species from the Footpad Abscess of a Black Vulture, Coragyps atratus
Strain BV1 was isolated from the exudate of the footpad abscess of a black vulture (Coragyps atratus). The colonies had a “fried-egg” appearance consistent with that of mycoplasmal species. Electron microscopic examination of the cells revealed irregular elongated or elliptical forms and smaller circular budding processes. Profuse growth was observed in Frey medium supplemented with 20% swine serum at 37°C in a humidified atmosphere of 10% CO2 and air. Typical of mycoplasma, strain BV1 required sterol for growth and catabolized glucose but did not hydrolyze arginie or urea. The guanine-plus-cytosine content of the DNA was 28 mol%. The organism demonstrated the ability to hemolyze, absorb onto, and agglutinate the erythrocytes from several animal species. Strain BV1 was serologically unrelated by the growth inhibition test to previously established Mycoplasma, Acholeplasma, Entomoplasma, and Mesoplasma species, as well as to strains belonging to these genera but not identified to species level. Moreover, BV1 had a 16S rRNA gene with a nucleotide sequence distinct from reported sequences of other mycoplasmas. This organism represents a new species for which the name Mycoplasma corogypsi is proposed. Strain BV1 (ATCC 51148T) is the type strain of Mycoplasma corogypsi sp. nov.
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Taxonomic Analysis of Thermophilic Strains of the Genus Methanobacterium: Reclassification of Methanobacterium thermoalcaliphilum as a Synonym of Methanobacterium thermoautotrophicum
More LessDNA reassociation, a comparative analysis of whole-cell protein patterns, and indirect immunofluorescence methods were used to determine the taxonomic relationships of thermophilic Methanobacterium strains. On the basis of the results dendrograms showing degrees of similarity among the organisms were constructed. The organisms studied are members of three different groups. One group, whose members exhibit 10 to 40% cross-hybridization, includes Methanobacterium wolfeii, “Methanobacterium defluvium” ADZ, and “Methanobacterium thermoflexum” IDZ. The second group contains Methanobacterium thermophilum MT (T = type strain) and Methanobacterium thermoaggregans (levels of DNA relatedness, 30 to 45%). The third group, whose members exhibit 65 to 99% cross-hybridization, includes Methanobacterium thermoautotrophicum ΔHT, F-1, and DV; Methanobacterium thermoformicicum Z-245T; and Methanobacterium thermoalcaliphilum AC60T. The combination of three independent taxonomic methods showed that the group of strains studied is phenotypically, genotypically, and antigenically diverse. The most distinct organisms are M. wolfeii, M. thermoaggregans, M. thermophilum, “M. defluvium,”“M. thermoflexum,” and M. thermoautotrophicum. These species names should be adopted. The results of a DNA-DNA hybridization study (level of hybridization, 99%), an immunological analysis (+3 reaction), and a protein similarity study (level of similarity, 75%) indicate that M. thermoalcaliphilum should be reclassified as a synonym of M. thermoautotrophicum.
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DNA Relatedness between Field Isolates of Mycoplasma F38 Group, the Agent of Contagious Caprine Pleuropneumonia, and Strains of Mycoplasma capricolum
DNA-DNA hybridization experiments were carried out in order to clarify the taxonomic relationships between the F38 group of caprine mycoplasmas, the established etiologic agents of classical contagious caprine pleuropneumonia, and Mycoplasma capricolum, an organism associated with septicemia, arthritis, and mastitis in goats and sheep. The taxonomic status of the F38 group has been uncertain, principally because of the serological, genomic, and other properties which it shares with M. capricolum. Tritium-labeled DNAs from the M. capricolum type strain (California kid) and from prototype strain F38 were hybridized with unlabeled DNAs from these two strains and from four other isolates belonging to each group. The results showed consistent DNA relatedness values of about 70% between the F38 and M. capricolum groups, compared with levels of relatedness of about 90 and 85%, respectively, for the strains within each group. In addition, the results of comparisons of these 10 strains in which growth inhibition and immunofluorescence tests were used confirmed the previously reported serological relationships between the two groups and reinforced other observations concerning their shared genomic and cell membrane characteristics, indicating that there is a close taxonomic relationship. However, as the 70% DNA relatedness values between the M. capricolum and F38 groups also indicate a degree of genomic difference inconsistent with a relationship at the species level, we conclude that our findings support previous proposals for classification of the F38 group as a subspecies of M. capricolum. In view of the prospective diagnostic problems, particularly those arising from the serological similarities of two putative subspecies, we believe that further studies should be performed to define additional phenotypic and genotypic properties that would allow more rapid and specific differentiation of the F38 group mycoplasmas.
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Phylogenetic Relationship of Chlamydia pneumoniae to Chlamydia psittaci and Chlamydia trachomatis as Determined by Analysis of 16S Ribosomal DNA Sequences
More LessThe 16S ribosomal DNA sequence of Chlamydia pneumoniae was determined and compared with the corresponding gene sequences of Chlamydia psittaci and Chlamydia trachomatis. C. pneumoniae has been reported to exhibit little chromosomal DNA homology with the other chlamydial species, and its phylogenetic relationships within the genus Chlamydia have not been described. A polymerase chain reaction was employed to determine the 16S rRNA gene sequence of C. pneumoniae. Ten primers from the C. psittaci sequences were used to amplify a C. pneumoniae template in overlapping segments of the gene. Sequence data for 1,554 bases indicated that the levels of homology of C. pneumoniae with C. psittaci and C. trachomatis were 96.19 and 94.07%, respectively. These data support the results of previous biochemical and developmental studies indicating that C. pneumoniae is more closely related to C. psittaci than to C. trachomatis.
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Stenotrophomonas, a New Bacterial Genus for Xanthomonas maltophilia (Hugh 1980) Swings et al. 1983
More LessIn consideration of the criticisms of the transfer of Pseudomonas maltophilia to the genus Xanthomonas proposed by J. Swings p. De Vos, M. Van den Mooter, and J. De Ley (Int. J. Syst. Bacteriol. 33:409-413, 1983), a new generic name is created for this taxon. The name Stenotrophomonas is here proposed for the new genus, which includes a single species, Stenotrophomonas maltophilia. This proposal restores the genus Xanthomonas to its former definition (J. Bradbury p. 199-210, in N. R. Krieg and J. G. Holt, ed., Bergey’s Manual of Systematic Bacteriology, 1984) The arguments on which this proposal is based are presented.
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Phylogenetic Relationship of Chlamydia pneumoniae to Chlamydia psittaci and Chlamydia trachomatis as Determined by Analysis of 16S Ribosomal DNA Sequences
More LessThe 16S ribosomal DNA sequence of Chlamydia pneumoniae was determined and compared with the corresponding gene sequences of Chlamydia psittaci and Chlamydia trachomatis. C. pneumoniae has been reported to exhibit little chromosomal DNA homology with the other chlamydial species, and its phylogenetic relationships within the genus Chlamydia have not been described. A polymerase chain reaction was employed to determine the 16S rRNA gene sequence of C. pneumoniae. Ten primers from the C. psittaci sequences were used to amplify a C. pneumoniae template in overlapping segments of the gene. Sequence data for 1,554 bases indicated that the levels of homology of C. pneumoniae with C. psittaci and C. trachomatis were 96.19 and 94.07%, respectively. These data support the results of previous biochemical and developmental studies indicating that C. pneumoniae is more closely related to C. psittaci than to C. trachomatis.
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Restriction Fragment Length Polymorphisms of rRNA as Genetic Markers To Differentiate Chlamydia spp.
More LessRestriction fragment length polymorphisms (RFLPs) of rRNA genes of Chlamydia spp. were analyzed. A Southern analysis of chromosomal DNA digests with cloned rRNA gene probes revealed the presence of one locus for rRNA genes on chromosomal DNAs in Chlamydia psittaci, Chlamydia pecorum, and Chlamydia pneumoniae and two loci in Chlamydia trachomatis. The RFLPs of rRNA genes were characteristic for each Chlamydia sp. DNA probes cloned from flanking regions of the rRNA genes of avian C. psittaci hybridized either with only C. psittaci or with only the avian and ovine abortion strains of C. psittaci. The RFLPs of rRNA genes and the flanking regions provide suitable genetic markers for differentiation of Chlamydia spp.
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Phylogenetic Position of Taylorella equigenitalis Determined by Analysis of Amplified 16S Ribosomal DNA Sequences
The 16S ribosomal DNA sequence of Taylorella equigenitalis (formerly Haemophilus equigenitalis), the causative organism of contagious equine metritis, was determined. A phylogenetic analysis of this sequence revealed a phylogenetic position of T. equigenitalis in the β subclass of the class Proteobacteria apart from the position of Haemophilus influenzae, which belongs to the γ subclass of Proteobacteria. A close phylogenetic relationship among T. equigenitalis, Alcaligenes xylosoxidans, and Bordetella bronchiseptica was detected; Spirillum volutans and Chromobacterium fluviatile (Iodobacter fluviatile) were in the same group but slightly removed. This relationship is surprising in view of the considerable differences in the G+C contents of the genomes of these bacteria.
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- Matters Relating To The International Committee On Systematic Bacteriology
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- Letters To The Editor
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- Errata
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Revised Taxonomy of the Class Mollicutes: Proposed Elevation of a Monophyletic Cluster of Arthropod-Associated Mollicutes to Ordinal Rank (Entomoplasmatales ord. nov.), with Provision for Familial Rank To Separate Species with Nonhelical Morphology (Entomoplasmataceae fam. nov.) from Helical Species (Spiroplasmataceae), and Emended Descriptions of the Order Mycoplasmatales, Family Mycoplasmataceae
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