- Volume 37, Issue 3, 1987

Five strains of a rapidly growing, orange, scotochromogenic Mycobacterium species were isolated from a marine sponge. Although they displayed similarities to M. aurum and M. parafortuitum, they proved to belong to a proposed new species, Mycobacterium poriferae sp. nov. (ATCC 35087). We distinguished the type strain of M. poriferae from that of M. aurum by its pattern of carbon sources used, acid production from carbon sources, and amidases, by its ability to tolerate 5% sodium chloride, its failure to use benzamide as sole nitrogen source, and its use of l-serine as sole carbon and nitrogen source. We distinguished the type strain of M. poriferae from that of M. parafortuitum by its pattern of carbon sources used, acid production from carbon sources, and amidases, by its ability to tolerate 5% sodium chloride, its failure to grow at 42°C, its strong pigmentation, its failure to reduce nitrate, its failure to tolerate 0.0250% hydroxylamine hydrochloride, its ability to use l-serine as dual carbon and nitrogen source, and its failure to use acetamide as dual carbon and nitrogen source. The pattern of mycolic acid derivatives produced by acid methanolysis of whole organisms was that of the group typified by M. avium and which contains α-mycolates, ketomycolates, and wax ester derivatives.

Mycoplasma strain 163K was isolated from the gills of a tench (Tinca tinca L.) with red disease. The cells are elongated, ovoid or flask shaped, consisting of a thicker body and a more slender part ending in a hemispherical terminal structure, that is apparently stabilized by a cytoskeleton. They are able to attach to inert surfaces and living cells. The most exciting property of the organisms is their fast gliding motion, which occurs on glass and plastic surfaces and which is not interrupted by resting periods. Growth occurs between 17 and 30°C, with an optimum at 25°C, in modified Hayflick medium with horse or bovine serum and in medium with swine serum. Acid is produced from several carbohydrates, but arginine and urea are not catabolized. The organism reduces tetrazolium chloride, produces film and spots and hydrogen peroxide, and is able to hemolyze and to adsorb to erythrocytes of several animal species. Indirect immunofluorescence test, growth inhibition, and metabolism inhibition tests indicate that the mycoplasma represents a new species for which the name Mycoplasma mobile is proposed. Strain 163K (ATCC 43663) is the type strain.

The taxonomic relationships of Moraxella urethralis, the Centers for Disease Control (CDC) group IVe, Taylorella equigenitalis, and other gram-negative bacteria were studied by deoxyribonucleic acid (DNA)-DNA polyacrylamide gel electrophoresis of cellular proteins, and serological, biochemical, and auxanographic analyses. A high relationship was detected between M. urethralis and the CDC group IVe strains. However, no relationship of M. urethralis and CDC group IVe with genuine Moraxella species was observed. We describe a new genus, Oligella, containing two species: Oligella urethralis (to accommodate Moraxella urethralis) and Oligella ureolytica (to accommodate CDC group IVe strains). The type species is Oligella urethralis, with type strain ATCC 17960T. The type strain of Oligella ureolytica is CDC C379 (ATCC 43534, CCUG 1465, LMG 6519). Oligella is a member of rRNA superfamily III, containing, e.g., the Pseudomonas acidovorans and Pseudomonas solanacearum complexes and Chromobacterium, Janthinobacterium, and Neisseria species. The closest relatives of Oligella species are Taylorella equigenitalis and the Alcaligenaceae family.

Streptoalloteichus hindustanus was originally described in 1978 as a genus and species of the family Actinoplanaceae. However, the taxonomic position of this organism should be transferred to a taxon that includes the genera Nocardiopsis, Actinosynnema, and Saccharothrix, based on the lowering of the taxonomic significance of sporangiumlike vesicles bearing flagellate spores and on a cell chemistry profile comprising the following: cell wall type III; a whole-cell sugar composition that is a variant of pattern C (presence of galactose, mannose, and rhamnose); menaquinones MK9(H6) and MK10(H6) as the major menaquinones; and phospholipid type P-II (presence of phosphatidylethanolamine). Chemotaxonomically, this organism is related to Saccharothrix australiensis, while it is differentiated by its Streptomyces-type morphology; i.e., spore chain formation at the tip of aerial hyphae, formation of two distinct types of spore chains, lack of mycelial fragmentation, and the capricious formation of sporangiumlike vesicles bearing flagellate spores. The type strain is C677-91T (ATCC 31217).

Cloned deoxyribonucleic acid (DNA) fragments, derived from a cosmid library of a Florida isolate of Xanthomonas campestris pv. citri 3041 were used to detect restriction fragment-length polymorphism among 93 strains of X. campestris, which make up 26 pathovars. DNA clones were radiolabeled and hybridized against Southern blot transfers of genomic DNAs of different strains of X. campestris digested with restriction endonucleases. Autoradiographs for DNA clones probed against genomic DNA revealed hybridization profiles which appeared to be highly conserved and unique for each pathovar tested. As a species, the population structure of X. campestris appeared basically clonal. Variability appeared to depend on the DNA probe used; some probes represented DNA loci which were highly variable, and some represented DNA loci which were highly conserved at the species level. By using more than one DNA probe, or by digesting the genomic DNAs with different restriction endonucleases, we were able to differentiate all of the strains of X. campestris described as belonging to a given pathovar. Differences among the pathovars were also confirmed by pathogenicity experiments on plants. Some phytopathogenic strains isolated from plants not previously described as susceptible to X. campestris could be grouped as being related to previously described pathovars.

Streptococci that produced acetoin and alkaline phosphatase, hydrolyzed arginine, and fermented trehalose but did not produce pyrrolidonylarylamidase or β-glucuronidase, split hippurate, or ferment ribose or glycogen were collected and compared. These streptococci were considered members of the unapproved species “Streptococcus milleri,” but most of them would fit the description of one of three approved species: Streptococcus anginosus, Streptococcus constellatus, or Streptococcus intermedius. Most were recent clinical isolates. Some hydrolyzed esculin and fermented lactose, while others did not. Some fermented mannitol and raffinose. Many were beta-hemolytic, and several reacted to antiserum of Lancefield group A, C, F, or G. From a total of 111 strains, 40 were selected for comparison of their deoxyribonucleic acid (DNA) base sequence similarities by DNA-DNA hybridization on membrane filters. All biotypes, hemolytic types, and serotypes were included, as well as the type strains of S. anginosus, S. constellatus, and S. intermedius, and Lancefield group F Streptococcus sp., plus two strains derived from Guthof’s “S. milleri” isolates. The results showed considerable genetic similarity within the group. DNA from most strains hybridized at a level of 70% or more of the homologous control, even under very stringent conditions. There was somewhat less homology between DNAs of some of the least reactive strains (lactose, mannitol, and esculin negative) and the most reactive strains (lactose, mannitol, and esculin positive). The Lancefield F strain and the type strains of S. anginosus and S. constellatus were genetically similar. Traits such as hemolysis and lactose fermentation could not be ascribed to plasmids. The results support the unification of these streptococci into a single species.

We used crossed immunoelectrophoresis, crossed-line immunoelectrophoresis, tandem-crossed immunoelectrophoresis, and crossed immunoelectrophoresis with an intermediate gel (containing antibodies) in studies of bacterial taxonomy. By these quantitative immunoelectrophoretic methods, as many as 90 antigens from a bacterial species can easily and reproducibly be analyzed for immunological cross-reactivity with antigens from other bacterial species without prior purification of antigens. The immunological cross-reactivity between various bacterial species is expressed as the matching coefficient (MC), i.e., the ratio between the number of cross-reactive antigens with another bacterial species and the total number of antigens of the reference bacterial species. The MC has been found to be significantly correlated with phenotypic similarities (biochemical characteristics) and nucleic acid hybridization results obtained with the taxa investigated so far. In general, closely related strains (serotypes within a species) have a high MC (≥0.90), while different species of the same genus have a lower MC (0.90 to 0.70 or lower in some cases); genera within a family have an intermediate MC (0.70 to 0.15), and distantly related taxa have a very low MC (≤0.15).

Transfer of Streptosporangium indianense Gupta 1965 to the genus Streptomyces Waksman and Henrici as Streptomyces indiaensis (Gupta 1965) comb. nov. is proposed on the basis of morphological and chemical properties. Spore chains such as those of typical streptomycetes were formed in the early stage of culture of S. indianense. True sporangia were not observed. Whole-cell hydrolysates contained ll-diaminopimelic acid. Compositions of cellular fatty acids and isoprenoid quinones were similar to those of streptomycetes. The type strain of Streptomyces indiaensis is JCM 3053T (= KCC A-0053, ATCC 33330, CBS 560.75, IFO 13964, and NCIB 9794).

Pseudomonas luteola Kodama et al. 1985 is a senior subjective synonym of Chryseomonas polytricha Holmes et al. 1986 (formerly group Ve-1). The type strain of P. luteola (JCM 3352) was 78% related to the type strain of C. polytricha (NCTC 11843). We propose the new combination Chryseomonas luteola for this organism. For Pseudomonas oryzihabitans Kodama et al. 1985 (formerly group Ve-2) we propose the new genus and new combination Flavimonas oryzihabitans. We were in the process of proposing a scientific name for group Ve-2 on the basis of an examination of 19 strains when the description of P. oryzihabitans was published. We have included the type strain of P. oryzihabitans for comparison and here present our results for all 20 strains. Each of the 20 strains was examined for 129 characteristics, including reactions in 58 enzyme tests (API ZYM system). These bacteria are rod shaped, aerobic, gram negative, yellow pigmented, motile by a single polar flagellum, and saccharolytic, but they do not produce cytochrome oxidase. The mean guanine-plus-cytosine content of the deoxyribonucleic acid of three representative strains was 63.6 ± 1.6 mol%. The deoxyribonucleic acid relatedness of 11 strains of group Ve-2 to the strain we would have proposed as the type strain, CL162/81 (= NCTC 11850), averaged 93% (hydroxyapatite method, 70°C). The type strain of P. oryzihabitans (JCM 2952) was 93% related to CL162/81. The relatedness of CL162/81 to the respective type strains of the five major ribosomal ribonucleic acid hybridization groups of the genus Pseudomonas and to the type strain of C. polytricha (formerly group Ve-1) was 5% or less. All strains, except the type strain, of F. oryzihabitans were isolated from human clinical specimens.

A total of 54 moderately halophilic vibrios, which were isolated from several salterns located in different areas of Spain, were examined by using a wide range of morphological, physiological, biochemical, and nutritional tests. The resulting data, together with data for four reference Vibrio costicola strains including the type strain V. costicola NCMB 701, and other marine species that were similarly examined, were compared by using several numerical taxonomic methods. There was a strong similarity between the 54 isolates and four reference strains of V. costicola that were isolated from cured meats. On the basis of these and other molecular data, including guanine-plus-cytosine content of the deoxyribonucleic acid and the plasmid content, we propose an amended description of this species.

Tests useful for the identification of Enterococcus strains were applied to a collection of isolates from animal intestines and to reference strains, all of which were capable of growth on 40% bile and in 6.5% NaCI. Most strains could be identified as known species, and their characteristics corresponded, with a few exceptions of minor importance, with those described for Enterococcus hirae, Enterococcus durans, Enterococcus mundtii, Enterococcus gallinarum, Enterococcus avium, and Enterococcus casseliflavus. However, some diagnostically important carbohydrate reactions of Enterococcus faecalis and Enterococcus faecium strains differed from those given in the species descriptions and in recent reports. Production of acid from d-raffinose andd-xylose by E. faecium varied with the host species from which the strains were isolated. E. durans and E. gallinarum were isolated only from poultry, whereas E. avium was found only in mammals.

A spiroplasma strain Ar-1343T (T = type strain), one of a large assemblage isolated from a mixed pool of Aedes stricticus and Aedes vexans mosquitoes in the French northern Alps, proved to be serologically unrelated to all currently recognized spiroplasma serogroups (I through XI). Strain Ar-1343T was examined with the criteria proposed by the International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Mollicutes for description of new mollicute species. These spiroplasmas were shown to belong to the class Mollicutes because of their membrane ultrastructure, their colony morphology, and their filtration patterns and to the family Spiroplasmataceae because of their helical morphology and motility. Growth in SP-4, M1A, or M1D media occurred at 20 to 32°C. Strain Ar-1343T was unable to grow in the absence of cholesterol in the medium. Glucose was fermented, and arginine was catabolized. The base composition (guanine plus cytosine) of the deoxyribonucleic acid of strain Ar-1343T was found to be 30 ± 1 mol%. On the basis of these findings, we propose that spiroplasma strain Ar-1343T, with the characteristics described here, should be recognized as a new species, Spiroplasma sabaudiense. Strain Ar-1343T has been deposited in the American Type Culture Collection, Rockville, Md. (ATCC 43303).

The 16S ribosomal ribonucleic acid catalog of Listeria denitrificans CIP 55134T was determined. The results confirmed previous evidence that this organism is not a member of the genus Listeria. Whereas L. monocytogenes is a member of the Bacillus-Clostridium subdivision of gram-positive bacteria, L. denitrificans is phylogenetically related to members of one of the several subbranches of the actinomycetes subdivision, defined by the genera Actinomycetes, Arthrobacter, Micrococcus, Cellulomonas, Stomatococcus, and Promicromonospora and allied taxa. The low similarity coefficients found between L. denitrificans and these genera, together with previously described chemotaxonomic properties, indicate that this species should be transferred to a new genus, Jonesia gen. nov., as Jonesia denitrificans comb. nov.

Five new species of motile and nonsporeforming anaerobic gram-negative rods that have deoxyribonucleic acids with guanine-plus-cytosine contents of 56 to 58 mol%, produce major amounts of propionic and acetic acids, and were isolated from the human gingival crevice are described: Selenomonas noxia with ATCC 43542 as the type strain, Selenomonas flueggei with ATCC 43531 as the type strain, Selenomonas infelix with ATCC 43532 as the type strain, Selenomonas dianae with ATCC 43527 as the type strain, and Selenomonas artemidis with ATCC 43528 as the type strain.

The validation of gas-vacuolated strains of methanosarcinae at the species level under the name of Methanosarcina vacuolata sp. nov. is presented, based on chemotaxonomic as well as morphological characteristics. M. vacuolata is shown to be related but distinct from the type species M. barkeri and the other species of freshwater methanosarcinae, M. mazei and M. thermophila.

The taxonomy of two groups (designated as 3 and 4) of alginate-degrading Bacillus strains formerly classified as Bacillus circulans was studied. Three new group 3 and two new group 4 strains were isolated from soil and studied. High intragroup deoxyribonucleic acid (DNA) relatedness values of 87 to 96% and low intergroup vlues of 27 to 36% indicated that these groups were genetically distinct taxa. Furthermore, low DNA complementarity values of 22 to 37% established that these two groups with guanine-plus-cytosine contents of 47 to 49 mol% were not closely related genetically to the type strains of recognized species with similar base contents. Low relatedness values were also measured between strains from the two groups and Bacillus pabuli, another alginate-degrading species. In general, negative reactions for growth at 50°C, for anaerobic growth, for casein hydrolysis, for nitrate reduction, and for acetylmethylcarbinol production differentiated the two groups from the named species. Growth in 0.001% lysozyme and utilization of citrate, fumarate, and succinate separated group 3 from group 4, which neither grew in lysozyme nor utilized the three organic acids. The data from the present and previous studies suggest that group 3 and 4 organisms are strains of two new species, for which are proposed the names Bacillus alginolyticus and Bacillus chondroitinus, respectively.

Arthrobacter variabilis (Müller) differs so much from the type species of the genus Arthrobacter, Arthrobacter globiformis, that it should not be retained within this genus. On the basis of biochemical and chemical criteria, it is proposed that Arthrobacter variabilis be reclassified in the genus Corynebacterium, as Corynebacterium variabilis comb. nov. The type strain is ATCC 15753 (NCIB 9455).

An unnamed and unclassified gas-vacuolated, facultatively anaerobic rod was characterized phenotypically and genotypically by deoxyribonucleic acid (DNA)-DNA hybridization. It most closely resembles members of the genus Aeromonas in the family Vibrionaceae. However, it differs phenotypically from the genus Aeromonas, and a low level of relatedness to its members was observed (11 to 22%). For these reasons, a new genus and species, Enhydrobacter aerosaccus, is proposed for this organism within the family Vibrionaceae.