- 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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