- Volume 35, Issue 1, 1985

A deoxyribonucleic acid-deoxyribonucleic acid hybridization study (spectrophotometric method) showed that the species Haemophilus avium Hinz and Kunjara 1977 is composed of three deoxyribonucleic acid homology groups; these three groups are genetically closer to Pasteurella multocida, the type species of the genus Pasteurella, than to Haemophilus influenzae, the type species of the genus Haemophilus. We propose to transfer the species Haemophilus avium to the genus Pasteurella as Pasteurella avium comb. nov. (type strain, ATCC 29546) and to change the circumscription of this taxon to include only strains that are unable to ferment l-arabinose, maltose, and mannitol and unable to hydrolyze o-nitrophenyl-β-d-galactopyranoside. The name Pasteurella volantium sp. nov. (type strain, NCTC 3438) is proposed for strains (formerly identified as H. avium) that are unable to ferment L-arabinose and are able to ferment maltose and mannitol and to hydrolyze o-nitrophenyl-β-d-galactopyranoside. A third unnamed species consists of former H. avium strains that are able to ferment l-arabinose.

An anaerobic bacterium which produced acetate from H2 and CO2 was isolated. The rod-shaped cells were not lysed by KOH, did not hydrolyze l-alanine-4-nitroanilide, and stained gram negative. However, the cell wall did not resemble a gram-negative wall in structure; it was comprised of two layers. The cells were motile by means of three or four peritrichous flagella. Yeast extract was required for both chemoorganotrophic and chemolithotrophic growth; yeast extract, glucose, maltose, or H2-CO2 could serve as a substrate for growth. Strain NOT-3T (T = type strain) grew best at 37°C and pH 7.6 to 7.8. The deoxyribonucleic acid base composition was 36.8 mol% guanine plus cytosine. Strain NOT-3 (= ATCC 35199) is named Acetoanaerobium noterae gen. nov., sp. nov. and is the type strain of this new species.

Seven strains of Flavobacterium thalpophilum which were isolated from clinical sources were compared with the type strains of Sphingobacterium mizutae and seven species of Flavobacterium. These 15 strains were examined for 11 biochemical characteristics; minor phenotypic variations were observed for the 7 strains of F. thalpophilum. All 15 strains were characterized by four major cellular fatty acids (13-methyltetradecanoate, 2-hydroxy-13-methyltetradecanoate, 3-hydroxy-15-methylhexadecanoate, and a monounsaturated 16-carbon straight-chain acid). Sphingophospholipid long-chain bases were detected in all strains of F. thalpophilum but were not detected in Flavobacterium balustinum, Flavobacterium breve, Flavobacterium indologenes, Flavobacterium meningosepticum, or Flavobacterium odoratum. F. thalpophilum, Flavobacterium multivorum, Flavobacterium spiritivorum, and S. mizuate contained major amounts of menaquinone 7 but no menaquinone 6, whereas F. balustinum, F. breve, F. indologenes, F. meningosepticum, and F. odoratum contained major amounts of menaquinone 6 but no menaquinone 7. The phenotypic and chemical characteristics of F. thalpophilum indicate a close taxonomic relationship with F. multivorum, F. spiritivorum, and S. mizutae.

Deoxyribonucleic acid relatedness studies (S1 nuclease method) showed that 25 strains of Haemophilus ducreyi from diverse geographic origins form a homogeneous deoxyribonucleic acid hybridization group. A total of 17 type or reference strains of established or proposed Haemophilus species were 0 to 6% related to H. ducreyi at the optimal temperature. These findings do not support the assignment of H. ducreyi to the genus Haemophilus.

Bacteria which utilize the xenobiotic compounds chloridazon, antipyrin, and pyramidon as sole carbon sources were isolated from various soil samples. The 22 strains isolated are similar with respect to morphological, physiological, biochemical, serological, and genetic properties. These bacteria are aerobic gram-negative rods or coccal rods (0.7 to 1.0 by 1.0 to 2.0 μm) that occur singly, in pairs, or in short chains and are nonmotile and nonsporeforming. Physiological and biochemical characteristics and susceptibility to antibiotics were determined. The strains need vitamin B12 as a growth factor; they are catalase positive and weakly oxidase positive and show slight H2S production. All of the other tests which we performed were negative. The nutritional spectrum is extraordinarily limited, with optimal growth on chloridazon, antipyrin, pyramidon, and l-phenylalanine. Most sugars, alcohols, amino and carboxylic acids, and ordinary complex media are not utilized. The bacteria are osmotically sensitive. They are a serologically uniform group of organisms, which are harmless to rats and rabbits. Their guanine-plus-cytosine contents range between 65 and 68.5 mol%. The chloridazon-degrading bacteria are characterized as a new genus, Phenylobacterium, with a single species, Phenylobacterium immobile. The type strain Phenylobacterium immobile strain E (= DSM 1986), is not closely related to any other gram-negative bacterium, as shown by a 16S ribosomal ribonucleic acid partial sequence analysis. This organism is a member of group I of the purple nonsulfur bacteria, but is phylogenetically isolated in this group. Phenylobacterium immobile is remotely related to Pseudomonas diminuta, Rhizobium leguminosarum, rhodopseudomonads, and Aquaspirillum itersonii. Like other members of this group, Phenylobacterium immobile contains 2,3-diamino-2,3-dideoxy-d-glucose in its lipopolysaccharide. The murein equals a normal murein from a gram-negative bacterium. All citric acid cycle enzymes are detectable in Phenylobacterium immobile.

The name Blastobacter natatorius is proposed for a new species of the genus Blastobacter. The type strain is strain UQM 2507, which was isolated from a freshwater swimming pool. B. natatorius is a yellow, gram-negative, oxidase-positive organism with rod-shaped or wedge-shaped cells with straight or slightly curved axes. Each cell has a simple mucilaginous holdfast at its nonreproductive pole by which it attaches to other cells or to solid surfaces to form rosettes. Reproduction occurs by asymmetric division (budding) of the mother cell to produce a spherical or ovoid daughter cell (bud). The free bud cells are motile, and each has a single polar flagellum. This organism is strictly aerobic and chemoorganotrophic. The guanine-plus-cytosine content of the deoxyribonucleic acid of the type strain is 65 mol%. The description of the genus Blastobacter is emended to include motile organisms and physiological information lacking because a culture of the type species, Blastobacter henricii, has never been isolated.

The results of deoxyribonucleic acid-deoxyribonucleic acid hybridization studies demonstrated a high degree of relatedness among “Haemophilus somnus,” “Haemophilus agni” “Histophilus ovis,” strains isolated from the preputial cavities of rams, and strains isolated from clinical cases of epididymitis in rams. The degree of relatedness between “Haemophilus somnus” and Actinobacillus seminis or Haemophilus influenzile was very small. From the results of our hybridization studies and previous studies on morphological characteristics, serology, and biochemical activity, we conclude that “Haemophilus somnus,” “Haemophilus agni,” and “Histophilus ovis” should be considered a single species. Inclusion in the genus Haemophilus does not appear to be appropriate. The data also suggest that bovine and ovine isolates may constitute two separate groups within the species. More extensive hybridization studies will be required for establishment of the definitive taxonomic position of this group of organisms.

Ten new Legionella species were characterized on the basis of biochemical reactions, antigens, cellular fatty acids, isoprenoid quinones, and deoxyribonucleic acid relatedness. Nine of the new species were isolated from the environment, and one, Legionella hackeliae, was isolated from a bronchial biopsy specimen obtained from a patient with pneumonia. The species all exhibited the following biochemical reactions typical of the legionellae: growth on buffered cysteine-yeast extract agar, but not on blood agar; growth requirement for cysteine; gram negative; nitrate negative; urease negative; nonfermentative; catalase positive; production of a brown pigment on tyrosine-containing yeast extract agar; liquefaction of gelatin; and motility. Legionella spiritensis was weakly positive for hydrolysis of hippurate; the other species were hippurate negative. Legionella cherrii, Legionella steigerwaltii, and Legionella parisiensis exhibited bluish white autofluorescence. Legionella rubrilucens and Legionella erythra exhibited red autofluorescence. The other species, L. spiritensis, L. hackeliae, Legionella maceachernii, Legionella jamestowniensis, and Legionella santicrucis did not autofluoresce bluish white or red. All species had cellular fatty acid contents qualitatively similar to those of previously described legionellae and had major amounts of ubiquinones with more than 10 isoprene units in the side chains. Each new species was serologically distinct from previously described Legionella species. As determined by the hydroxyapatite method at 60°C, two strains of L. maceachernii were 100% related, and four strains of L. cherrii were 94 to 99% related. The other new species were represented by single strains. The levels of relatedness of the new species to each other and to previously described legionellae ranged from 1 to 67%. L. maceachernii, L. jamestowniensis, and L. hackeliae were less than 25% related to other species. L. rubrilucens and L. erythra, and two red-autofluorescing species, were about 60% interrelated. L. spiritensis (a non-autofluorescing species) was 34% related to L. rubrilucens. L. santicrucis was 64% related to Legionella sainthelensi. The three bluish white-autofluorescing species, L. parisiensis, L. cherrii, and L. steigerwaltii, were most closely related to other bluish white-autofluorescing species, especially Legionella bozemanii, Legionella dumoffii, Legionella gormanii, and “Legionella anisa” (35 to 67%).

Deoxyribonucleic acid relatedness studies (SI nuclease method) showed that 60 strains proposed as Budvicia aquatica form a homogeneous deoxyribonucleic acid hybridization group. Three strains labeled Budvicia-like were 9 to 22% related to B. aquatica. A total of 74 strains representing known species and genera in the Enterobacteriaceae were 0 to 8% related to B. aquatica. These findings support designation of Budvicia aquatica as a new genus and new species. This new species in the family Enterobacteriaceae is composed of strains which produce H2S; hydrolyze urea and o-nitrophenyl-β-d-galactopyranoside; do not produce acid from trehalose, d-mannose, glycerol, sucrose, maltose, and d-melibiose; do not decarboxylate lysine, ornithine, or arginine; do not produce phenylalanine deaminase; and have complex growth factor requirements. The guanine-plus-cytosine content of the deoxyribonucleic acid is 46 mol%. The type strain is strain 20186HG01 (= ATCC 25567).

We describe the comparative ultrastructures of bile-sensitive Bacteroides species which were isolated from oral cavities and ferment xylose and arabinose. Reference strains Bacteroides buccae ATCC 33574T(T = type strain), Bacteroides capillus ATCC 33690Tand ATCC 33691, and Bacteroides pentosaceus NP333Tand WPH61 and Bacteroides sp. strains ES42 and ES57 all had an extra surface layer (S-layer) outside the outer membrane. No S-layer was detected in Bacteroides oris ATCC 33573Tand ATCC 27518, Bacteroides oralis, Bacteroides veroralis, or Bacteroides sp. strains ES2759 and ES2834. The deoxyribonucleic acid guanine-plus-cytosine contents and both the malate dehydrogenase and glutamate dehydrogenase mobilities of the strains with an S-layer were identical. We suggest that the oral pentose-fermenting Bacteroides isolates with an S-layer may belong to the same species.

Deoxyribonucleic acid base composition, deoxyribonucleic acid-deoxyribonucleic acid hybridization, lipid, and biochemical studies were performed with Enterococcus faecium NCDO 1258 (S= Snell strain R) and other atypical Enterococcus faecium strains from pigs and chickens in an attempt to clarify their taxonomy. Our results indicate that these strains constitute a new species, for which the name Enterococcus hirae sp. nov. is proposed. The type strain of Enterococcus hirae is strain NCDO 1258.

We determined the sensitivities of 33 strains and variants of cyanobacteria to infection by the cyanophage LPP-1 archaetype, five LPP-1 serotypes, six LPP-2 serotypes, and 8 new LPP isolates. The LPP-1 archaetype and LPP-1 serotypes have different host ranges on strains of LPP group B.

The taxonomy of the genus Gluconobacter has undergone many changes during the past 40 years. Based on phenotypic properties, this genus has changed from numerous species to one species containing five subspecies and then to a single species, Gluconobacter oxydans, with no subspecies. The present study was designed to test the validity of this latter view. Nucleotide sequence similarites were determined for 54 strains of Gluconobacter by using an S1 nuclease procedure. Three distinct deoxyribonucleic acid homology groups were obtained. The average level of relatedness among these groups was 16%. Homology group 1 contained 32 strains and included the type strain of G. oxydans and the type strains of all previously recognized subspecies. Homology group II contained 12 strains that had an intragroup homology level of 44 to 87% (average, 65%) with reference strain IFO 3264. Homology group III contained six strains with an average intragroup homology level of 86% with reference strain IFO 3276a. Reference strains IFO 3264 and IFO 3276 were previously recognized as G. oxydans. The remaining four strains of Gluconobacter had from 0 to 23% homology with reference strains used to delineate the three homology groups. Although these data show that the genus Gluconobacter is composed of at least three species, they also support elimination of the previously designated subspecies. Three isolates implicated in pink disease of pineapples were shown either not to be gluconobacters or to be mixed with gluconobacters. The occurrence of colony variants within many of the Gluconobacter strains is described, and the significance of this observation is discussed.

We studied 170 strains of Actinomadura, Nocardiopsis, and related taxa. The overall similarities of the test strains were determined by comparing 120 unit characters, using the simple matching and pattern coefficients. Clustering was achieved by using the unweighted pair group method with averages. Cluster composition was not markedly affected by the coefficient used or by test error (estimated at 4.9%). The numerical data, together with results from previous chemical and genetic studies, showed that the genus Actinomadura is markedly heterogeneous. Strains received as Actinomadura fell into two large aggregate groups. One, Actinomadura sensu stricto, contained Actinomadura citrea, Actinomadura coerulea, Actinomadura coeruleoviolacea, Actinomadura cremea, Actinomadura livida, Actinomadura madurae, Actinomadura malachitica, Actinomadura pelletieri, and Actinomadura verrucosospora. Actinomadura kijaniata was not studied, but other genetic data suggest that it should also be included in this group. The other aggregate group encompassed Actinomadura ferruginea, Actinomadura pusilla, Actinomadura roseola, Actinomadura roseoviolacea, Actinomadura rubra, and Actinomadura salmonea. The representatives of Actinomadura helvata, Actinomadura libanotica, Actinomadura luteofluorescens, Actinomadura spadix, and Actinomadura spiralis studied formed distinct clusters separate from both of the aggregate groups. Nocardiopsis strains formed a cluster clearly distinguished from the clusters containing the Actinomadura strains.

A new subspecies, Staphylococcus aureus subsp. anaerobius, is described on the basis of a study of 84 strains isolated from young sheep affected by the so-called “abscess disease.’’ The strains of this new subspecies grow well under anaerobic conditions, but not at all or only very weakly under aerobic conditions. They are catalase and benzidine negative and form small unpigmented colonies. Anaerobically they produce l-lactic acid from glucose. The chemical composition of the cell wall, the results of deoxyribonucleic acid-deoxyribonucleic acid hybridization experiments, and the immunological relationships among l-lactate dehydrogenases demonstrated that these strains are closely related to Staphylococcus aureus. The type strain is strain MVF-7(= ATCC 35844).

We prepared antisera against purified inducible β-galactosidases from Lactobacillus murinus ATCC 35020, Lactobacillus bulgaricus ATCC 11842T (T = type strain), and Lactobacillus helveticus ATCC 15009T and used these antisera to measure the levels of immunological relatedness between the reference enzymes and β-galactosidases from various lactobacilli. Crude extracts from 11 species of lactobacilli, 7 wild-type strains isolated from Tafí cheese (Tucumán, Argentina), and 4 biotypes isolated from the guts of rats were tested against β-galactosidase antisera from L. murinus, L. bulgaricus, and L. helveticus. Pairwise comparisons between cross-reacting crude extracts were done by using double-diffusion assays. These comparisons permitted the recognition of antigenic specificity groups based on the β-galactosidases of various nomenspecies of lactobacilli; some of the species gave bands of nonidentity. The results of microcomplement fixation experiments in which we used two of the three antisera supported the results obtained with the double-diffusion assays and provided quantitative estimations of the antigenic relationships within the various β-galactosidases. From the combined results obtained with the antisera, a composite phylogenetic map of the antigenic relationships among the β-galactosidases from lactobacilli was prepared. This map agreed with the maps for the following two other constitutive enzymes: fructose diphosphate aldolase (J. London and K. Kline, Bacteriol. Rev. 37:453–478, 1973) and d-(–)-lactic dehydrogenase (F. Gasser and C. Gasser J. Bacteriol. 106:113–125, 1971).

Violet-pigmented bacteria isolated from the estuarine waters of Waccasassa Bay, Fla., were found to have biochemical and physiological characteristics very similar to those of Alteromonas luteoviolacea Gauthier 1982. All isolates tested had sheathed flagella, which seem to exclude them from the genus Alteromonas Baumann et al. 1972 as presently described. We suggest altering the genus description to include strains with this characteristic.

The type strain of Bacteroides pentosaceus Shah and Collins and the type strain of Bacteroides buccae Holdeman et al. have more than 80% deoxyribonucleic acid homology with the type strain of Bacteroides capillus Kornman and Holt, and therefore the three names are subjective synonyms. Since B. buccae Holdeman et al. was the first validly published name applied to this taxon, B. capillus and B. pentosaceus are later synonyms of B. buccae.

We describe a new species of purple nonsulfur bacteria, which has the ability to grow under photoautotrophic growth conditions with sulfide as an electron donor and shows the characteristic properties of Rhodobacter species (i.e., ovoid to rod-shaped cells, vesicular internal photosynthetic membranes, bacteriochlorophyll a and carotenoids of the spheroidene series as photosynthetic pigments). In its physiological properties this new species is particularly similar to the recently described species Rhodobacter adriaticus, but it shows enough differences compared with R. adriaticus and the other Rhodobacter species to be recognized as a separate species. In honor of Hans Veldkamp, a Dutch microbiologist, the name Rhodobacter veldkampii sp. nov. is proposed.