- Volume 47, Issue 3, 1997
Volume 47, Issue 3, 1997
- Original Papers Relating To Systematic Bacteriology
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Tsukamurella tyrosinosolvens sp. nov.
AbstractChemotaxonomic and 16S ribosomal DNA sequence analyses of four bacterial isolates from blood cultures from patients with cardiac pacemaker implants and sputa of patients with chronic lung infections clearly demonstrated that these bacteria belong to the genus Tsukamurella. DNA-DNA hybridization data, as well as the physiological characteristics of the isolates, indicate that they are closely related and belong to a single species that differs from previously described members of the genus Tsukamurella. The name Tsukamurella tyrosinosolvens sp. nov. is proposed for these isolates, and the new species is represented by strain IMMIB D-1397T (= DSM 44234T). Strain IMMIB D-1397T exhibits 53.4, 53.5, and 54.7% DNA-DNA relatedness to Tsukamurella paurometabola DSM 20162T, Tsukamurella inchonensis DSM 44067T, and Tsukamurella pulmonis DSM 441 42T, respectively.
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Description of Two New Thermophilic Desulfotomaculum spp., Desulfotomaculum putei sp. nov., from a Deep Terrestrial Subsurface, and Desulfotomaculum luciae sp. nov., from a Hot Spring
AbstractSix strains of thermophilic, endospore-forming, sulfate-reducing bacteria were enriched and isolated from 2.7 km below the earth’s surface in the Taylorsville Triassic Basin in Virginia. The cells of these strains were motile rods that were 1 to 1.1 µm in diameter and 2 to 5 µm long. The cells grew by oxidizing H2, formate, methanol (weakly), lactate (incompletely, to acetate and CO2), or pyruvate (incompletely) while reducing sulfate to sulfide; acetate did not serve as a catabolic substrate. Thiosulfate or sulfite could replace sulfate as an electron acceptor. The results of a phylogenetic analysis of the 16S rRNA gene indicated that these strains belong to the genus Desulfotomaculum, but are distinct from previously described Desulfotomaculum species. Thus, we propose a new species, Desulfotomaculum putei, for them, with strain TH-11 (= SMCC W459) as the type strain. The results of our phylogenetic analysis also indicated that strain SLTT, which was isolated from a hot spring and has been described previously (T. M. Karnauchow, S. F. Koval, and K. F. Jarrell, Syst. Appl. Microbiol. 15:296–310, 1992), is also a member of the genus Desulfotomaculum and is distinct from other species in this genus. We therefore propose the new species Desulfotomaculum luciae for this organism; strain SLT (= SMCC W644) is the type strain of D. luciae.
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Thermococcus hydrothermalis sp. nov., a New Hyperthermophilic Archaeon Isolated from a Deep-Sea Hydrothermal Vent
AbstractAn extremely thermophilic archaeon, strain AL662T, was isolated from a deep-sea hydrothermal vent located on the East Pacific Rise at a latitude of 21°N. This strain is a strictly anaerobic coccus, and its cells range from 0.8 to 2 p.m in diameter. The optimum temperature, pH, and Sea Salt concentration for growth are 85°C, 6, and 20 to 40 g/liter, respectively. Strain AL662T grows preferentially on proteolysis products, on a mixture of 20 amino acids, and on maltose in the presence of elemental sulfur. The membrane lipids consist of di- and tetraether glycerol lipids. The DNA G+C content is 58 mol%. Sequencing of the 16S rRNA gene showed that strain AL662T belongs to the genus Thermococcus. On the basis of hybridization results, we propose that this strain should be placed in a new species, Thermococcus hydrothermalis.
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Helicobacter rodentium sp. nov., a Urease-Negative Helicobacter Species Isolated from Laboratory Mice
More LessAbstractA spiral-shaped bacterium with bipolar, single, nonsheathed flagella was isolated from the intestines of laboratory mice. The organism grew at 37 and 42°C under microaerobic and anaerobic conditions, did not hydrolyze urea, was weakly positive for catalase and oxidase, reduced nitrate to nitrite, did not hydrolyze indoxyl acetate or hippurate, and was resistant to cephalothin and nalidixic acid. This is the first urease-negative, murine Helicobacter spp. isolated from intestines. Also, Helicobacter pullorum and this bacterium are unique among the genus Helicobacter in having nonsheathed flagella. The new bacterium appears to be part of the normal intestinal flora; although its pathogenic potential is unknown, this organism was also isolated from scid mice with diarrhea that were co-infected with Helicobacter bilis. On the basis of 16S rRNA gene sequence analysis data and biochemical and phenotypic criteria, the new organism is classified as a novel helicobacter, for which we propose the name Helicobacter rodentium. The type strain is MIT 95-1707 (= ATCC 700285).
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Comparison of the 16S Ribosomal DNA Sequences from the Intracellular Agents of Proliferative Enteritis in a Hamster, Deer, and Ostrich with the Sequence of a Porcine Isolate of Lawsonia intracellularis
More LessAbstractProliferative enteritis is an enteric disease that affects a variety of animals. The causative agent in swine has been determined to be an obligate intracellular bacterium, Lawsonia intracellularis, related to the sulfatereducing bacterium Desulfovibrio desulfuricans. The intracellular agents found in the lesions of different animal species are antigenically similar. In addition, strains from the pig, ferret, and hamster have been shown to be genetically similar. In this study we performed a partial 16S ribosomal DNA sequence analysis on the intracellular agent of proliferative enteritis from a hamster, a deer, and an ostrich and compared these sequences to that of the porcine L. intracellularis isolate. Results of this study indicate that the intracellular agents from these species with proliferative enteritis have high sequence similarity, indicating that they are all in the genus Lawsonia and that they may also be the same species, L. intracellularis.
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Comparison of a New Insertion Element, IS1407, with Established Molecular Markers for the Characterization of Mycobacterium celatum
More LessAbstractGenomic analyses of 18 Mycobacterium celatum strains obtained from different patients in three countries (United States, United Kingdom, and France) were performed; the methods used in this study were restriction fragment length polymorphism (RFLP) analysis, pulsed-field gel electrophoresis (PFGE) analysis, and PCR restriction analysis (PRA) of the hsp-65 gene. A new insertion sequence, IS1407 (GenBank accession no. X97307), belonging to the IS256 family, was identified in M. celatum type 1 strains and was characterized by sequencing. When a probe for Mycobacterium xenopi IS1395-like sequences was used, the RFLP analysis of M. celatum type 1 strains revealed that they contained three or four copies of IS1407 in identical genomic positions, while this element was absent in all M. celatum type 2 strains. PFGE performed with three different endonucleases revealed a unique large restriction fragment (LRF) pattern for M. celatum type 1 strains, whereas the LRF patterns obtained for M. celatum type 2 strains were polymorphic. Moreover, PFGE of nondigested genomic DNA revealed extrachromosomal elements in M. celatum type 2. The type strain of M. celatum type 3 could not be differentiated from M. celatum type 1 strains on the basis of the results of the RFLP analysis, the PFGE analysis, and the PRA of 1S1407. In this study we confirmed that M. celatum types 1 and 2 represent distinct genomic clusters and that the molecular markers in M. celatum type 2 exhibit greater polymorphism than the molecular markers in M. celatum type 1.
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Genotypic and Phenotypic Diversity within Streptococcus anginosus
More LessStreptococcus anginosus is one of the three species currently included in the “anginosus group” of oral or viridans streptococci. In this study 21 strains of S. anginosus were examined in order to determine whether this species, as currently defined, is sufficiently heterogeneous to warrant further subdivision. Phenotypic strain characterization was carried out by performing biochemical tests with a commercial system (Rapid ID32 STREP kit; bioMerieux), by performing tests to determine hyaluronidase production, hemolysis on blood agar, and gliding motility on chocolate agar, by determining Lancefield groups, and by comparing whole-cell polypeptide patterns obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Variations in genotype were determined by studying 16S-23S rRNA intergenic spacer size polymorphisms by PCR amplification, by ribotyping, and by performing DNA-DNA base pairing studies. S. anginosus was found to be heterogeneous at both the species and intraspecies (subspecies) levels. Beta-hemolytic Lancefield group C strains that did not produce hyaluronidase formed a DNA homology group that was separate from the majority of the S. anginosus strains; the members of this group produced a 380-bp intergenic spacer PCR product, exhibited distinct ribotypes, produced an atypical SDS-PAGE pattern, and represented a previously undescribed species in the anginosus group. Two other strains (ATCC 9895 and 1007-77) remained ungrouped as determined by DNA-DNA hybridization and thus represented additional centers of variation at the species level. Hyaluronidase-producing, beta-hemolytic, Lancefield group C strains produced the same atypical SDS-PAGE pattern as beta-hemolytic Lancefield group C strains that did not produce hyaluronidase but differed from the latter organisms by producing a 600-bp intergenic spacer PCR product. In addition, both DNA homology data and ribotyping data suggested that these strains comprise a subspecies of S. anginosus. With the notable exception of the beta-hemolytic Lancefield group C strains that did not produce hyaluronidase, strains ATCC 9895 and 1007-77, and the beta-hemolytic Lancefield group C hyaluronidase-producing strains mentioned above, the strains studied formed a closely related group within which some additional genotypic and phenotypic heterogeneity was observed. The latter group included both strains that fermented mannitol and strains that did not ferment mannitol, as well as strains that exhibited so-called gliding motility. Although no clear-cut division of these organisms was possible, our results indicate that strain NCTC 10713 may not be the most suitable type strain for S. anginosus. We concluded that S. anginosus strains exhibit sufficient heterogeneity to warrant division at both the species and subspecies levels, although insufficient numbers of strains belonging to the putative new taxa have been characterized to allow formal taxonomic proposals to be made.
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Caloramatorproteoclasticus sp. nov., a New Moderately Thermophilic Anaerobic Proteolytic Bacterium
More LessAbstractA new moderately thermophilic proteolytic anaerobe, strain UT, was isolated from mesophilic granular methanogenic sludge. The cells were spore-forming, motile rods that were 0.4 μm wide and 2.4 to 4 μm long and stained gram negative. Electron micrographs of thin sections revealed the presence of an atypical gram-positive cell wall. Optimum growth occurred at 55°C and at pH values between 7.0 and 7.5, with a doubling time of 30 min. The DNA base ratio of guanine plus cytosine was 31 mol%. The bacterium fermented proteins mainly to acetate, hydrogen, formate, and branched-chain fatty acids. Several amino acids, including glutamate, aspartate, arginine, histidine, threonine, methionine, and branched-chain amino acids, were also utilized. Glutamate was degraded to acetate, formate, hydrogen, and alanine. In addition, the strain degraded carbohydrates, including glucose, fructose, mannose, cellobiose, and starch, to acetate, ethanol, formate, lactate, and hydrogen. The results of a 16S rRNA sequence analysis phylogenetically placed strain UT in the low-guanine-plus-cytosine-content subgroup of the gram-positive phylum. We propose to classify the described strain in the genus Caloramator as a new species, Caloramator proteoclasticus. The type strain of C. proteoclasticus, strain U, has been deposited in the Deutsche Sammlung von Mikroorganismen as strain DSM 10124.
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Reclassification of the Crenarchaeal Orders and Families in Accordance with 16S rRNA Sequence Data
More LessAbstractA phylogenetic analysis of all validly published members of the Crenarchaeota, including several new isolates from our laboratory, suggests three orders within this archaeal kingdom. The Thermoproteales consist of both the rod-shaped, hyperthermophilic, neutrophilic representatives of the Thermoproteaceae and the members of the new family Thermofilaceae. The Sulfolobales harbor all thermoacidophilic, coccoid organisms. The neutrophilic, hyperthermophilic cocci are members of a new order tentatively named “Igneococcales.” This order comprises two families, the Desulfurococcaceae, characterized by maximal growth temperatures of up to 100°C, and the new family Pyrodictiaceae, for which optimal growth occurs at temperatures above 100°C.
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Inter- and Intraspecific Genetic Analysis of the Genus Saccharomonospora with 16S to 23S Ribosomal DNA (rDNA) and 23S to 5S rDNA Internally Transcribed Spacer Sequences
More LessIn order to clarify interspecific relationships and to investigate the intraspecific phylogenetic structure of the genus Saccharomonospora, 16S to 23S ribosomal DNA (16S-23S) and 23S to 5S ribosomal DNA (23S-5S) internally transcribed spacers (ITSs) were used for sequence analyses. The 16S-23S and 23S-5S ITSs from 22 Saccharomonospora strains were amplified by PCR and directly sequenced. The average levels of nucleotide similarity of the 16S-23S and 23S-5S ITSs for the four valid species were 87.6% ± 3.9% and 83% ± 2.2%, respectively. For the most part, intraspecific sequence differences were not found in the two ITSs; the only exception was Saccharomonospora glauca K194, which differed from other S. glauca strains by 1 bp in the 23S-5S ITS. The Saccharomonospora viridis strains had a smaller 16S-23S ITS region than the other strains, which may be useful for differentiating these organisms from other Saccharomonospora species. The characteristics of the two ITS regions make them more useful than 16S rRNA sequences as a tool for defining and identifying Saccharomonospora strains. However, Saccharomonospora azurea K161Thad two types of 23S-5S ITSs; rrnB, separated by Xho I digestion, had two additional nucleotides inserted between positions 52 and 55. Most of the 16S-23S and 23S-5S ITS sequences of S. azurea K161Tand strains of “Saccharomonospora caesia” were identical; the only exception was rrnB in S. azurea K161T. The lengths and levels of sequence divergence of the two ITSs of Saccharomonospora sp. strain K180 were different from the lengths and levels of sequence divergence of the ITSs of other species. These findings suggest that a taxonomic revision of the genus Saccharomonospora is necessary. Two trees based on 16S-23S and 23S-5S ITS sequences revealed distinct interspecific relationships in the genus Saccharomonospora.
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Psychroserpens burtonensis gen. nov., sp. nov., and Gelidibacter algens gen. nov., sp. nov., Psychrophilic Bacteria Isolated from Antarctic Lacustrine and Sea Ice Habitats
More LessPsychrophilic, yellow-pigmented, seawater-requiring bacteria isolated from the pycnocline of meromictic Burton Lake and from sea ice cores obtained in the Vestfold Hills (68°S, 78°E) in eastern Antarctica were characterized. Phenotypic analysis showed that the strains isolated formed two distinct taxa. The first taxon included nonmotile, nutritionally fastidious strains that were isolated from the pycnocline of Burton Lake. The cells of these strains were morphologically variant, ranging from vibrioid to ring shaped to coiled and filamentous; in addition, the strains were unable to metabolize carbohydrates or polysaccharides and had DNA G+C contents of 27 to 29 mol%. The strains of the second taxon, which were isolated from sea ice cores and from ice algal biomass, were saccharolytic, exhibited rapid gliding motility, were rodlike to filamentous, and had DNA G+C contents of 36 to 38 mol%. A 16S ribosomal DNA (rDNA) sequence analysis revealed that the two Antarctic taxa formed related but distinct lineages within the [Flexibacter] maritimus rRNA branch of the family Flavobacteriaceae. The levels of 16S rDNA sequence similarity between the taxa were 90.5 to 91.3%, while the levels of similarity to other members of the [F.] maritimus rRNA branch were 85 to 90%. The whole-cell lipid profiles of the Antarctic strains were mainly comprised of branched and unbranched monounsaturated C15to C17fatty acids. The presence of significant levels of the lipids a15:1 ω 10c and a17:1 ω 7c appeared to be useful biomarkers for the new Antarctic taxa and for differentiating these organisms from other members of the family Flavobacteriaceae. On the basis of polyphasic taxonomic data we propose that the new taxa are novel bacterial species designated Psychroserpens burtonensis gen. nov., sp. nov. (type strain, ACAM 188) and Gelidibacter algens gen. nov., sp. nov. (type strain, ACAM 536).
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Use of Ribotyping To Distinguish Bordetella bronchiseptica Isolates
More LessA total of 113 Bordetella bronchiseptica strains, isolated from 11 different host species worldwide, were characterized by ribotyping with restriction enzyme Pvu II. Sixteen distinct ribotypes were identified, and each ribotype contained five to seven restriction fragments ranging in size from 1.8 to 5.6 kb. Approximately 88% of the swine isolates were identified as ribotype 3 strains. Isolates from dogs also displayed little variation; 74.1% were found to be ribotype 4 strains. Strains obtained from the remaining nine host species belonged to 15 different ribotypes. There was no association between geographic location and ribotype. The technique which we used may be useful for epidemiologic studies in which the transmission of B. bronchiseptica, both within and between species, is investigated.
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16S rRNA Gene Sequence of Rubrobacter radiotolerans and Its Phylogenetic Alignment with Members of the Genus Arthrobacter, Gram-Positive Bacteria, and Members of the Family Deinococcaceae
More LessThe nearly complete sequence of the 16S rRNA gene of an extremely highly radiotolerant bacterium, Rubrobacter radiotolerans(reclassified from Arthrobacter radiotolerans based on chemical characteristics), was determined by PCR amplification of the genomic DNA followed by cloning of the amplified gene and sequencing by the dideoxynucleotide method. The sequence was aligned with the sequences of members of the genus Arthrobacter and also with the sequences of representatives of the gram-positive bacteria having high G+C contents and the family Deinococcaceae(radioresistant micrococci and their relatives). The results of our phylogenetic analysis confirmed that R. radiotolerans is not a member of the Arthrobacter group and thus supported the previous reclassification. Moreover, although it is radioresistant and has a high G+C content, R. radiotolerans is more closely related to the gram-positive bacteria with high G+C contents than to the radioresistant members of the Deinococcaceae.
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Actinomyces europaeus sp. nov., Isolated from Human Clinical Specimens
Ten strains of a hitherto undescribed catalase-negative, facultatively anaerobic, coryneform bacterium were isolated or collected by workers at three European clinical bacteriology laboratories or reference centers. These strains were isolated from humans, and most came from abscess material. Biochemical and chemotaxonomic characterization revealed that the strains belonged to the genus Actinomyces. The phenotypic features of the 10 strains were incompatible with the descriptions of the previously established Actinomyces species. A comparative 16S rRNA gene sequence analysis demonstrated that the previously undescribed strains constitute a new line in the genus Actinomyces. The name Actinomyces europaeus sp. nov. is proposed for these clinical isolates. The type strain is CCUG 32789A.
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rRNA Sequences and Evolutionary Relationships among Toxic and Nontoxic Cyanobacteria of the Genus Microcystis
A primary-structure analysis of the 16S rRNA gene was performed with 10 strains representing five described and one unidentified species of the genus Microcystis. The phylogenies determined illustrate the evolutionary affiliations among Microcystis strains, other cyanobacteria, and related plastids and bacteria. A cluster of 10 strains that included hepatotoxic isolates identified as Microcystis aeruginosa formed a monophyletic group. However, the genus Microcystis appeared to be polyphyletic and contained two strains that clustered with unicellular cyanobacteria belonging to the genus Synechococcus. The clustering of related Microcystis strains, including strains involved in the production of the cyclic peptide toxin microcystin, was consistent with cell morphology, gas vacuolation, and the low G+C contents of the genomes. The Microcystis lineage was also distinct from the lineage containing the unicellular genus Synechocystis and the filamentous, heterocyst-forming genus Nostoc. The secondary structure of a Microcystis 16S rRNA molecule was determined, and genus-specific sequence signatures were used to design primers that permitted identification of the potentially toxic cyanobacteria belonging to the genus Microcystis via DNA amplification.
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Discrimination of Members of the Family Pasteurellaceae Based on Polyamine Patterns
More LessIn a study of the classification of members of the family Pasteurellaceae, the polyamine patterns of 101 strains were analyzed. These strains included the type strains of species belonging to the genera Actinobacillus, Haemophilus, and Pasteurella and additional strains of selected species, as well as numerous unnamed strains. Members of the genus Actinobacillus sensu stricto were characterized by the presence of 1,3-diaminopropane as the predominant compound. In the majority of the species of the genus Haemophilus sensu stricto 1,3-diaminopropane was also the major compound in the polyamine pattern. In contrast, Haemophilus intermedius subsp. gazogenes and Haemophilus parainfluenzae were characterized by high levels of 1,3-diaminopropane, cadaverine, and putrescine. These results confirmed the findings of Dewhirst et al. (F. E. Dewhirst, B. J. Paster, I. Olsen, and G. J. Fraser, Zentralbl. Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt. 1 Orig. 279:35–44, 1993), who demonstrated that H. parainfluenzae is phylogenetically only distantly related to the type species of the genus Haemophilus, Haemophilus influenzae. The phylogenetic diversity of the genus Pasteurella sensu stricto determined by Dewhirst et al. was also reflected to some extent by different polyamine patterns. The common characteristics found in Pasteurella multocida, Pasteurella canis, Pasteurella dagmatis, Pasteurella stomatis, and Pasteurella sp. strain B were high levels of putrescine and spermidine and the presence of the unusual triamine sym-norspermidine. Pasteurella avium, Pasteurella gallinarum, and Pasteurella volantium contained high concentrations of 1,3-diaminopropane and spermidine. Pasteurella langaa contained only high concentrations of 1,3-diaminopropane, and Pasteurella anatis was characterized by the presence of 1,3-diaminopropane as the predominant compound and high levels of putrescine and spermidine. Our data demonstrate that polyamine patterns are useful for discrimination within the family Pasteurellaceae.
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Spiroplasma lampyridicola sp. nov., from the Firefly Beetle Photuris pennsylvanicus
Spiroplasma strain PUP-1Twas isolated from the gut fluids of a firefly beetle (Photuris pennsylvanicus) collected in Maryland. Cells of the strain were shown by dark-field microscopy to be helical, motile filaments. Ultrastructural examination by electron microscopy revealed filamentous cells bounded by a single cytoplasmic membrane and no evidence of a cell wall. The cells were not sensitive to 500 U of penicillin per ml and grew under aerobic conditions in M1D, SP-4, and M-2 broth formulations, as well as in conventional mycoplasma medium. The doubling times at 15, 20, 25, and 30°C were 83.1, 32.0, 14.9, and 9.8 h, respectively. Suboptimal growth occurred at 37°C, and no growth was apparent in cultures maintained at 10 or 40°C. The organism required cholesterol for growth and produced acid from glucose, fructose, and trehalose; arginine and urea were not hydrolyzed. The results of previous serological analyses of strain PUP-1Tindicated that the organism was not related to the then currently established Spiroplasma species or group representatives, and the organism was classified as the representative of group XIX. Additional testing of strain PUP-1Tagainst recently recognized Spiroplasma species or group representatives by both metabolism inhibition and deformation tests confirmed the unique serological status of the organism. The guanine-plus-cytosine content of the DNA was 26 ± 1 mol%, and the genome size was 1,375 kbp. These values clearly differentiate strain PUP-1Tfrom group XXI strain W115, with which it cross-reacted reciprocally at a low level in deformation and metabolism inhibition tests. We propose that strain PUP-1 (= ATCC 43206) should be recognized as the type strain of a new species, Spiroplasma lampyridicola.
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Spiroplasma chrysopicola sp. nov., Spiroplasma gladiatoris sp. nov., Spiroplasma helicoides sp. nov., and Spiroplasma tabanidicola sp. nov., from Tabanid (Diptera: Tabanidae) Flies
Four spiroplasma strains, DF-1T, TG-1T, TABS-2T, and TAUS-1T, all of which were isolated from deerflies or horseflies (Diptera: Tabanidae), were serologically distinct from previously described spiroplasma species, groups, and subgroups. Strain DF-1Toriginated from a Maryland deerfly (Chrysops sp.); strain TG-1Twas isolated from a Maryland horsefly (Tabanus gladiator); strain TAUS-1Toriginated from a member of the Tabanus abdominalis-limbatinevris complex of horseflies collected in Maryland; and strain TABS-2Twas isolated from a horsefly (Tabanus abactor) collected in Oklahoma. Cells of all of the strains appeared to be helical and motile when they were examined by dark-field microscopy. Cells of strain DF-1Tgrowing in M1D medium were short helices with less than six turns; the helical cells of the other strains were long and usually had six or more turns. The short cells of strain DF-1Tpassed through 450- and 300-nm filter pores with no reduction in titer, but the longer cells of the other strains were partially retained by 450-nm-pore-size filters. Electron microscopic examination of all of the strains revealed wall-less cells surrounded only by a single cytoplasmic membrane. All of the strains grew well in SP-4 liquid media and in conventional mycoplasma or M1D media supplemented with horse or fetal bovine serum. Strains TABS-2T, TAUS-1T, and DF-1Trequired serum or sterol for growth, but strain TG-1Twas able to grow in the absence of serum or sterol. The optimum temperatures for growth of the four strains varied from 30 to 32°C, and growth occurred at 10 to 37°C. All of the strains catabolized glucose but did not hydrolyze urea. Only strain DF-1Thydrolyzed arginine. The guanine-plus-cytosine contents of the DNAs of the strains were: DF-1T, 29 ± 1 mol%; TG-1T, 26 ± 1 mol%; TABS-2T, 27 ± 1 mol%; and TAUS-1T, 26 ± mol%. The genome sizes of strains DF-1Tand TAUS-1Twere 1,270 and 1,375 kbp, respectively. Strain DF-1 (= ATCC 43209), the representative of spiroplasma subgroup VIII-2, is designated the type strain of a new species, Spiroplasma chrysopicola. We also propose that strain TG-1T(= ATCC 43525T), the designated representative of group XXIII, should be placed in a new species, Spiroplasma gladiatoris. In addition, group XXXII spiroplasma strain TABS-2 (= ATCC 51746) is designated the type strain of Spiroplasma helicoides sp. nov., and group XXXIII representative strain TAUS-1 (= ATCC 51747) is designated the type strain of another new species, Spiroplasma tabanidicola.
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Spiroplasma montanense sp. nov., from Hybomitra Horseflies at Northern Latitudes in North America
Spiroplasma strain HYOS-1Twas isolated from a tabanid fly, Hybomitra opaca. The organism was serologically distinct from other spiroplasma species, groups, and subgroups and was recently designated the representative of spiroplasma group XXXI. The cells of strain HYOS-1T, as determined by light microscopy, were long motile helices. Electron microscopic examination revealed wall-less cells delimited by a single membrane. The cells passed through 450- and 300-nm filter pores with a 10-fold reduction in titer, but failed to pass through 100-nm pores. Strain HYOS-1Tgrew very well in most conventional medium formulations for spiro-plasmas or other mollicutes. The organism grew at temperatures ranging from 5 to 41°C, and the optimum temperature was 32°C. The doubling time at the optimum temperature was 0.7 h, one of the shortest values obtained for members of the genus Spiroplasma. The strain catabolized glucose and hydrolyzed arginine but not urea. Growth of the organism was stimulated by cholesterol and serum, but the strain was nevertheless able to grow in the absence of sterols or serum. The guanine-plus-cytosine content of the DNA was about 28 ± 1 mol%, and the genome size was 1,225 kbp. On the basis of the experimental results reported here and previously reported data, group XXXI strain HYOS-1 (= ATCC 51745) is designated the type strain of a new species, Spiroplasma montanense.
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Staphylococcus lutrae sp. nov., a New Coagulase-Positive Species Isolated from Otters
More LessPhenotypic and phylogenetic studies were performed with three strains of a catalase-positive, gram-positive, coccus-shaped bacterium isolated from otters. The results of a 16S rRNA gene sequence analysis demonstrated that these strains represent a hitherto unknown subline within the genus Staphylococcus. Based on the results of the phylogenetic analysis and phenotypic criteria, we propose that these bacteria should be classified as members of a new species, Staphylococcus lutrae. The type strain of S. lutrae is DSM 10244.
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