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Volume 28,
Issue 2,
1978
Volume 28, Issue 2, 1978
- Contributors To Systematic Microbiology
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- Original Papers Relating To Systematic Bacteriology
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Grouping of Staphylococci on the Basis of Their Bacteriolytic-Activity Patterns: A New Approach to the Taxonomy of the Micrococcaceae
More LessAs a further development of our previous finding that heterogeneous bacteriolytic activities are produced by virtually all staphylococcal strains, several different conditions were tested for their selective and differentiating interferences with the bacteriolytic activity of some staphylococcal strains. Such a study has indicated the possibility of resolving different patterns of lytic activity among staphylococci and has resulted in the preparation of an assay system, composed of eight testing media, for lytic enzymes. A total of 1,054 staphylococcal strains has been tested on these media and found to fall into six “lyogroups,” each characterized by a specific pattern of bacteriolytic activity. The novelty and taxonomic relevance of analyzing the bacteriolytic activity of staphylococci and the possibility of extending the proposed method to bacteria other than staphylococci are discussed.
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Grouping of Staphylococci on the Basis of Their Bacteriolytic-Activity Patterns: A New Approach to the Taxonomy of the Micrococcaceae
More LessA number of characters (colony features, biochemical and metabolic properties, susceptibility to antimicrobial agents) were determined on 1,054 staphylococci previously placed in six “lyogroups” on the basis of bacteriolytic activity. A great deal of uniformity was found among the characteristics of strains of the same lyogroup. Virtually 100% agreement was found between four of the six lyogroups and four nomen species or biotypes: i.e., lyogroup I was found to equate with Staphylococcus aureus; lyogroups II and III were found to equate with the recently described species S. simulans and S. capitis, respectively; and lyogroup V was found to equate with S. epidermidis sensu stricto. The occurrence of a relationship between the bacteriolytic activities of staphylococci and their cell wall compositions is shown. Such a correlation may be an expression of a specific mechanism of cell wall growth. The taxonomic significance of bacteriolytic activity and the relationship of this criterion and the resultant classificatory scheme to other criteria and schemes are discussed.
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Intra- and Intergeneric Similarities of Chromobacterium and Janthinobacterium Ribosomal Ribonucleic Acid Cistrons
More LessWe prepared hybrids between 14C-labeled ribosomal ribonucleic acid (rRNA) from either Chromobacterium violaceum NCTC 9757 or Chromobacterium lividum NCTC 9796 and deoxyribonucleic acid (DNA) from a great variety of named gram-negative bacteria, including many type and reference strains. Each hybrid was described by two parameters: (i) Tm(e) , the temperature at which 50% of the hybrid was denatured; (ii) the percent rRNA binding, the amount (micrograms) of [14C]rRNA duplexed in stringent conditions per 100 μg of filter-fixed homologous or heterologous DNA. Each taxon occupied a definite area on the rRNA similarity maps. All C. lividum and C. violaceum strains formed tight separate clusters around their neotype strains. The rRNA parameters of both taxa were about as different from each other as was the case for many genera. Both taxa are phenotypically and genotypically (by DNA-DNA hybridization) likewise very different. We proposed to elevate each cluster to genus rank as C. violaceum and Janthinobacterium lividum. The following strains were misnamed and belonged in neither genus: “C. lividum” = “C.” folium NCTC 10590 and 10591; “C. lividum” GA; “C.” marismortui ATCC 17056; “Pseudomonas” (“Chromobacterium”) iodinum ATCC 9897, 15728, and 15729; “C.” indicum-rubrum (now Serratia marcescens) NCTC 2847; and “C.” viscosum (now Corynebacterium sp.) NCTC 2416. The rRNA cistrons of Chromobacterium and Janthinobacterium resembled most closely those of Pseudomonas section II and III, the authentic Alcaligenes, Bordetella bronchiseptica, the H2-oxidizing Alcaligenes eutrophus and Alcaligenes paradoxus, and Comamonas percolans NCIB 8193. These taxa displayed a number of phenotypical similarities. We suggested that all these taxa are the closest taxonomic relatives of Chromobacterium and Janthinobacterium.
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Numerical Classification of Rhodococcus (Formerly Gordona) Organisms Recently Isolated from Sputa of Patients: Description of Rhodococcus sputi Tsukamura sp. nov.
More LessA total of 116 strains of Rhodococcus (formerly Gordona) recently isolated from sputa of patients with pulmonary disease were submitted to numerical classification together with 61 reference strains belonging to the genera Rhodococcus, Nocardia, and Mycobacterium. The numerical classification was carried out by two methods, one using 109 characters and another using 98 characters. Almost similar results were obtained by these two methods, and the following clusters were distinguished: (i) Rhodococcus bronchialis (synonym: Gordona bronchialis); (ii) R. sputi sp. nov.; (iii) R. rubropertinctus (synonym: G. rubropertincta); (iv) R. rhodochrous (synonym: G. rhodochroa); (v) R. aurantiacus (synonym: G. aurantiaca); (vi) R. lentifragmentus (synonym: G. lentifragmenta); (vii) Nocardia rubra strains received from M. Ridell; (viii) N. asteroides; (ix) N. farcinica; (x) Mycobacterium fortuitum. Clusters i to vii were combined into one large cluster, which was considered as the genus Rhodococcus (synonym: Gordona). In addition, two (method I) or four (method II) new clusters could be observed among the cluster Rhodococcus. Thus, it was considered that the Rhodococcus organisms form a genus that contains a number of species. G. terrae and G. rosea were incorporated into the cluster of R. rubropertinctus (G. rubropertincta), and were considered to be reduced to varieties of the species R. rubropertinctus. The largest one of the new clusters was named Rhodococcus sputi sp. nov. Tsukamura. Its definition is presented in this study. A proposal of changing the generic name from Gordona to Rhodococcus was made.
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Deoxyribonucleic Acid Homology and Taxonomy of the Genus Bacillus
More LessThe taxonomic relationships among 56 strains of 16 species of the genus Bacillus were studied by deoxyribonucleic acid (DNA)-DNA hybridization. In general, no significant DNA homology was detected between two strains of different species, except for a group of species consisting of B. subtilis, B. amyloliquefaciens, B. licheniformis, and B. pumilus and for another group of species including B. cereus and B. thuringiensis. Species of the former group were related, but they were independent of each other as their DNA homologies were 19% or less. The DNA homology indexes of three strains of B. thuringiensis to B. cereus T, so far tested, showed high DNA homologies (54 to 80%). This fact indicates that B. thuringiensis should be an identical species to B. cereus. The intraspecific DNA homology indexes of 16 strains of B. pumilus were 51% or more to strain IFO 12092 as the standard, and those of 10 strains of B. coagulans were 76 to 113% to strain ATCC 7050 as the standard. Thus, the species identification of B. pumilus and B. coagulans by the conventional taxonomic method was well in accord with the DNA homology data. On the other hand, significant heterogeneities were suggested among the strains of B. circulans and among those of B. sphaericus by the DNA homology data of the three and five strains so far tested, respectively. Although B. lentus was described to be closely related to B. firmus in Bergey’s Manual (8th ed., 1974), the interspecific DNA homology index between these two species was 3%. It was concluded that these two species are independent.
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Streptococcus raffinolactis Orla-Jensen and Hansen, a Group N Streptococcus Found in Raw Milk
More LessThe properties of the lactate dehydrogenases, percent guanine plus cytosine in the deoxyribonucleic acid (DNA), and DNA/DNA hybridization studies have shown that three strains of group N streptococci do not belong to either Streptococcus lactis or Streptococcus cremoris. The biochemical properties of the three strains were published about 25 years ago, and at that time the strains were not assigned to any species. The three strains are here identified as members of Streptococcus raffinolactis Orla-Jensen and Hansen.
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Isoniazid Susceptibility as a Criterion for the Differentiation of Mycobacterial Species from Other Mycolic Acid-Containing Taxa
More LessOne hundred twenty-five strains of mainly fast-growing mycobacteria and representatives of related taxa were screened for susceptibility to 50, 100, and 200 μg of isoniazid per ml. Over 80% of the mycobacterial strains were susceptible to 200 μg of isoniazid per ml, with the resistant ones restricted to the species Mycobacterium album, M. aurum, M. chelonei, M. farcinogenes, and M. fortuitum. With a single exception, representatives of the taxa Bacterionema, Corynebacterium, Micromonospora, Nocardia, Rhodococcus, Streptomyces, and Gordona aurantiaca were resistant to the concentrations of isoniazid tested. The susceptibility of most fast-growing mycobacteria to high concentrations of isoniazid seems to provide the basis of a useful test to distinguish the majority of these organisms from related actinomycete and coryneform bacteria.
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Bacteroides ureolyticus, a New Species to Accommodate Strains Previously Identified as “Bacteroides corrodens, Anaerobic”
More LessThe name Bacteroides ureolyticus is proposed for a species to accommodate strains of gram-negative, urease-positive, anaerobic, corroding rods previously incorrectly referred to as Bacteroides corrodens. The organisms are catalase negative and reduce nitrate. In peptone-yeast-glucose medium, growth is enhanced by the addition of fumarate and formate, and succinate is the major end product. Conventional carbohydrate fermentation tests are negative. The oxidase test is positive and is markedly inhibited by azide. Absorption bands of reduced cytochromes are seen by visual spectroscopy at 550 nm (cytochrome c) and 555-560 nm (b-type cytochrome); no cytochrome α band was detected. Most strains hydrolyze gelatin and casein. Strains with very weak proteolytic activity are also encountered. The cells are nonflagellated but may show “twitching motility.” Electron micrographs of five of seven strains reveal polar pili. Strains that lack pili do not produce spreading colonies or show twitching motility. The guanine-plus-cytosine content of the deoxyribonucleic acid is in the range of 28.0 to 30.0 mol%. Strain NCTC 10941 is designated as the type strain.
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Revised Description, from Clinical Strains, of Flavobacterium breve (Lustig) Bergey et al. 1923 and Proposal of the Neotype Strain
More LessFlavobacterium breve (Lustig 1890) Bergey et al. 1923 was originally meagerly characterized in comparison with the wide range of characterization tests now available, and subsequent descriptions of this species have been few and incomplete. No type strain was designated for F. breve, and no neotype appears to have been proposed. Six strains from a collection of 1,700 gram-negative, nonfermen-tative clinical strains submitted to the National Collection of Type Cultures for computer-assisted identification during the last 10 years and one strain (ATCC 14234) maintained in a culture collection as F. breve conformed to the original and subsequent descriptions of F. breve. One of these seven strains, CL88/76 (= NCTC 11099), is herein proposed as the neotype strain of F. breve. These seven strains have been examined in a large number of biochemical tests and in their susceptibility to a range of antimicrobial agents in order to provide a revised description of the species by which it may be more easily recognized in clinical material. One of our clinical strains died before its susceptibility could be determined, but the remaining five, together with ATCC 14234, were resistant to carbenicillin and gentamicin, and the five clinical strains examined were also resistant to several other antimicrobial agents generally useful in the treatment of infections caused by gram-negative, nonfermentative bacteria. This suggests that infections due to this species could prove to be difficult to treat.
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Corynebacterium pilosum and Corynebacterium cystitidis, Two New Species from Cows
More LessTwo strains of corynebacteria were isolated from cows that showed signs of cystitis and pyelonephritis. According to the results of numerical-analysis and deoxyribonucleic acid homology studies, these strains differed from the known species of Corynebacterium parasitic on or pathogenic to humans and/or other animals. These strains are regarded as belonging to two new species, for which the names C. pilosum and C. cystitidis are proposed. The type strains of these species are 46 Hara (= ATCC 29592) and 42 Fukuya (= ATCC 29593), respectively.
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Two New Species of Alteromonas: Alteromonas espejiana and Alteromonas undina
More LessA total of 32 amino-acid-requiring, gram-negative, polarly flagellated, marine, rod-shaped bacteria were submitted to an extensive phenotypic characterization. Numerical analysis of the data grouped the strains into three clusters, one of which was identified as Alteromonas haloplanktis. The strains in the remaining clusters had properties characteristic of the genus Alteromonas and were phenotypically distinct from each other as well as from the previously characterized species of this genus. One cluster was designated Alteromonas espejiana sp. nov. (type strain, 261 [= ATCC 29659]), and the other was designated Alteromonas undina sp. nov. (type strain, 272 [= ATCC 29660]).
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Comparative Study of the Total Cellular Fatty Acids of Staphylococcus Species of Human Origin †
More LessThe cellular fatty acid compositions of 100 strains representing 10 different species of Staphylococcus were analyzed by gas-liquid chromatography. Results indicated that the fatty acid compositions of the various species were qualitatively similar with the exception of that of S. warneri, which contained a C22 fatty acid component. The species differed quantitatively with respect to relative percentages of fatty acid components. The major fatty acids were C15Br, C17Br, C16, C18, and C20. S. warneri and S. saprophyticus can be differentiated in most cases from all of the other species studied by the presence of significantly higher percentages of C20 and C16 fatty acids, respectively. Other species or groups of related species could also be separated on the basis of combinations of fatty acid components. These findings corroborate the newly proposed scheme of classification of the staphylococci, but the particular usefulness of gas-liquid chromatography of fatty acids for the identification of clinically significant, coagulase-negative staphylococci from humans is primarily limited to S. saprophyticus and possibly S. haemolyticus. However, application of the method may be useful in clarifying further the phenetic relationships of certain species and may have some ecological value.
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Comparative Zone Electrophoresis of Enzymes in the Genus Bacillus
More LessExtracts from 88 strains representing a wide variety of Bacillus species were examined by zone electrophoresis, followed by specific stains for each of 11 enzyme activities, and were also analyzed for deoxyribonucleic acid (DNA) buoyant density. Strains representing B. megaterium, B. cereus, B. laterosporus, B. firmus, B. alvei, B. polymyxa, and B. macerans exhibited unique patterns of enzyme migrations for each species. Most strains of B. subtilis and B. amyloliquefaciens also displayed characteristic enzyme migration patterns for each species. However, one strain, B. subtilis KA63, resembled B. amyloliquefaciens in DNA composition and enzyme types. It was previously reported to resemble B. amyloliquefaciens in DNA-DNA homology. Another group of strains, which was homogeneous and distinctive in electrophoretic enzyme types, included two strains of B. subtilis subsp. niger and one of B. subtilis subsp. globigii. Yet another group contained the type strains for B. licheniformis, B. pulvifaciens, and B. coagulans. A second B. coagulans strain was quite different from that in the above-mentioned group or from any other Bacillus strain examined. Strains of B. pumilus fell into two distinct biotypes according to the electrophoretic data; culturally, they comprised a homogeneous set. Representatives of B. thuringiensis, B. bombycis, and B. entomocidus were not distinguishable from B. cereus by these data.
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Taxonomy of the Bacteroides
More LessStrains of the “Bacteroides fragilis group” and other saccharolytic intestinal bacteroides were delineated by deoxyribonucleic acid homology. Of the 340 strains studied, about 90% belonged to any 1 of 10 major homology groups: The named species B. fragilis, B. vulgatus, B. distasonis, B. thetaiotaomicron, B. ovatus, B. uniformis, and B. eggerthii, a described group temporarily designated “subsp. a,” and groups “3452-A” and “T4-1.” Organisms belonging to the two latter groups had phenotypic similarities with and had not been previously differentiated from B. distasonis. Thirty more strains had no homology with the above-mentioned groups and appeared to be very heterogeneous when compared among themselves. There were two subgroups within the B. fragilis homology group. These subgroups had about 65 to 70% intergroup homology and about 80 to 90% intragroup homology. The B. ovatus and B. thetaiotaomicron homology groups had similar levels of heterogeneity, but they were more complex in that they appeared to have greater numbers of subgroups. The rest of the homology groups were quite homogeneous, usually having 80 to 90% intragroup homology.
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Taxonomy of the Bacteroides
More LessTwo hundred ninety strains of Bacteroides, each belonging to B. fragilis, B. thetaiotaomicron, B. ovatus, B. uniformis, B. vulgatus, B. distasonis, or B. eggerthii, or to one of three unnamed taxa designated “B. fragilis subsp. a,” “3452-A,” and “T4-1,” were used to compare the patterns of 34 phenotypic traits with deoxyribonucleic acid (DNA) homology groupings. The responses for all but 13 of the traits were very similar for all of the DNA homology groups. The responses for the 13 traits (the production of indole, catalase, and hydrogen, and acid production from arabinose, cellobiose, melibiose, melezitose, raffinose, rhamnose, ribose, salicin, sucrose, and trehalose) varied between the various homology groups. Similarity coefficients, for which all of the strains were compared with the DNA reference strains, were calculated. Similarity coefficient averages of strains within a given DNA homology group with the DNA reference strains of that group ranged form 83 to 94%. Organisms having about 60 to 65% DNA homology to each other were in most cases also phenotypically very similar. In a couple of instances, however, homology groups with strains having 72 to 95% intrastrain homology were quite variable with respect to certain phenotypic traits. There was no general pattern between levels of intergroup homologies and phenotypic properties. For example, B. eggerthii and the “subsp. a” homology groups, having about 50% intergroup homology, had average intergroup phenotypic similarity values of 65 and 69%, whereas B. ovatus and B. thetaiotaomicron homology groups, having only about 35% intergroup DNA homology, had intergroup similarity values that ranged from 86 to 93%. Similarity coefficients between homology groups having low levels of DNA homology ranged from 38 to 78%. Response probability values for the phenotypic traits were estimated for strains belonging to each of the DNA homology groups. The probability values obtained with 20 to 40 strains were not altered significantly by including more strains. A test identification matrix was constructed using the probability values of the 13 phenotypic traits listed above. On the basis of these traits, 93% of the strains could be assigned to the correct homology group if groups having 60% or greater intragroup homology were considered to be phenotypically the same.
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Deoxyribonucleic Acid Relatedness of Proteus and Providencia Species
Deoxyribonucleic acid reassociation was used to determine relatedness among protei and providenciae and between these organisms and other members of the family Enterobacteriaceae. The results indicate the following. (i)Proteus mirabilis, Prot. morganii, and Providencia stuartii are homogeneous species. (ii) Prot. vulgaris, Prot. rettgeri, and Prov. alcalifaciens each contain more than one deoxyribonucleic acid relatedness group. (iii) A group of urea-positive strains previously called Prot. rettgeri biogroup 5 are, in fact, members of Prov. stuartii. (iv)Prot. myxofaciens is a valid species. (v) Protei are only distantly related to all other Enterobacteriaceae. Taxonomic revisions consistent with these observations are discussed. These include a proposal to place the currently recognized species of Proteus into one of three genera: The genus Proteus, containing Prot. mirabilis Hauser, Prot. vulgaris Hauser, and Prot. myxofaciens Cosenza and Podgwaite; a separate genus Morganella Fulton, for Prot. morganii (Winslow et al.) Yale; and the genus Providencia containing Prov. alcalifaciens (De Salles Gomes) Ewing, Prov. stuartii (Buttiaux et al.) Ewing, and Prov. rettgeri comb, nov.
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Rhodocyclus purpureus gen. nov. and sp. nov., a Ring-Shaped, Vitamin B12-Requiring Member of the Family Rhodospirillaceae
More LessThe dominant phototrophic bacterium in a water sample from a purplish red waste-water lagoon was isolated in pure culture. The individual cells of this organism were half-ring-shaped and ring-shaped before cell division; the half-rings were 0.6 to 0.7 μm wide and 2.5 to 3.0 μm long. Open or compact coils of variable length were also formed. Acetate, pyruvate, and cyclohexane carboxylate were the carbon sources best utilized by this organism; vitamin B12, p-aminobenzoic acid, and biotin were required as growth factors. The photosynthetic pigments produced by the organism are bacteriochlorophyll αp and carotenoids of the rhodopinal series. The new bacterium was facultatively aerobic and was unable to photooxidize sulfide or thiosulfate to sulfur or sulfate. It belongs to the family Rhodospirillaceae and is described herein as a member of a new genus, Rhodocyclus. The name proposed for this new species is Rhodocyclus purpureus. The type strain of R. purpureus is “Ames” 6770 (= DSM 168).
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Mycoplasma sualvi, a New Species from the Intestinal and Urogenital Tracts of Pigs
More LessEight similar Mycoplasma isolates were obtained from rectal, intestinal, and vaginal specimens from pigs. Two indistinguishable clones of one isolate (May-field) were examined in detail. Although closely resembling the cattle organism Mycoplasma alvi in cultural and biochemical properties, the Mayfield clones differed from this species in morphological appearance and deoxyribonucleic acid base composition. The clones were serologically distinct from M. alvi and 58 currently recognized Mycoplasma species and subspecies, including all of those having similar biochemical properties or the same host. The Mayfield strain clone B (NCTC 10170) is designated the type strain of a new species, M. sualvi, which includes five of the seven other Mycoplasma isolates from pigs. The remaining two isolates appear to be serologically dissimilar from the Mayfield strain.
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Cytophaga aquatilis sp. nov., a Facultative Anaerobe Isolated from the Gills of Freshwater Fish
More LessA facultatively anaerobic, gram-negative, gliding bacterium was isolated from the gills of freshwater fish. Its deoxyribonucleic acid base composition (33.7 mol% guanine plus cytosine), lack of microcysts or fruiting bodies, cell size (0.5 by 8.0 μm), and hydrolysis of carboxymethylcellulose and chitin place it in the genus Cytophaga. This aquatic cytophaga is differentiated from other cytophagas by its fermentation of carbohydrates, proteolytic capabilities, and a number of additional physiological and biochemical tests. The organism was compared to other similar isolates reported from fish, and it appears to belong to a new species, for which the name Cytophaga aquatilis is proposed. The type strain of C. aquatilis, N, has been deposited with the American Type Culture Collection under the accession number 29551.
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