- Volume 28, Issue 1, 1978

The lipoquinones and associated respiratory functions of 34 cultures representing 17 bacterial species hitherto assigned to the genus Actinobacillus or Pasteurella were investigated. Strains representing recognized Actinobacillus or Pasteurella species regularly contained desmethylmenaquinones and were capable of anaerobic electron transport with fumarate as a terminal acceptor when grown in oxygen-limited cultures. On aeration, the majority of the strains produced ubiquinones in addition to desmethylmenaquinones, with inter- and intraspecific variations. The use of lipoquinone patterns and features indicating fumarate respiration, such as stimulation of oxygen-limited growth by fumarate, in the classification and identification of Actinobacillus, Pasteurella, and phenotypically similar organisms is discussed.

Respiratory quinones and the ability to use fumarate as a terminal electron acceptor in anaerobic respiration were investigated in 49 bacterial strains representing a variety of conventional Flavobacterium or Cytophaga species. The organisms examined were subdivided into two categories according to their quinones. (i) Ubiquinones are used by the neotype strain of Flavobacterium aquatile and by cultures representing F. acidificum, F. capsulatum, F. devorans, F. halmephilum, and some unnamed Flavobacterium species. (ii) Menaquinones are produced by both typical Cytophaga strains and many so-called Flavobacterium or “Flavobacterium/Cytophaga” cultures. Several members of category ii exhibited low to medium reduced nicotinamide adenine dinucleotide-fumarate reductase activities when grown in unaerated complex media supplemented with fumarate. In addition, with F. meningosepticum, “group IIb” organisms, and a strain of F. odoratum, the yields of oxygen-limited growth were markedly increased by fumarate, indicating an energetic use of fumarate respiration. On the basis of these findings, restriction of the genus Flavobacterium to “low-guanine-plus-cytosine” organisms containing ubiquinones and resembling F. aquatile is proposed. The incorporation of some former “flavobacteria” into a natural group of organisms containing menaquinones and placement in the vicinity of the C. hutchinsonii guanine-plus-cytosine ratio are discussed.

The aim of the present study was to solve the taxonomic problems of haemophili of porcine origin hitherto classified as Haemophilus parahaemolyticus Pittman 1953 or H. pleuropneumoniae Shope 1964. Forty-seven Haemophilus strains isolated mainly from pneumonic lesions in pigs and of different geographic origin were subjected to biochemical and serological examinations. All strains were V dependent and urease positive, and the majority were beta-hemolytic. For comparison, nine beta-hemolytic and V-dependent strains isolated from humans were examined. The porcine strains were divisible into two distinct groups on the basis of their biochemical characteristics, hemolytic activity, and deoxyribonucleic acid base composition. Both groups of porcine organisms were separated from the beta-hemolytic strains of human origin, previously named H. parahaemolyticus, by a significant number of biochemical and cultural characteristics and thus should not be classified with these. Following the rule of priority of proposed names, the correct name for the major group of porcine organisms is Haemophilus pleuropneumoniae (Matthews and Pattison 1961) Shope 1964. The species is described in detail and a neotype strain (Shope 4074) is proposed. The minor group of porcine strains possibly deserves specific recognition, but a recommendation with regard to this matter is deferred pending further studies.

A survey of 272 free-living rhizobia strains showed that a majority of the Rhizobium japonicum and unclassified Rhizobium isolates are capable of both growth and acetylene reduction on a Casamino Acids-minimal medium. Two other taxonomic categories of Rhizobium were discernible: R. meliloti and R. trifolii, which grow but do not reduce acetylene; and R. leguminosarum, which fails to grow on the minimal medium. Sixty-nine strains were acetylene reducers obtained from root nodules of the plant hosts: Albizzia, Cassia (partridge pea), Baptisia (wild indigo), Crotalaria, Ulex (thornbroom), Indigofera, Lespedeza, Desmodium (Florida beggarweed), Arachis (peanut), Phaseolus, Strophostyles, Stizolobium (velvet bean), Pueraria (kudzu), Vigna (cowpea), Erythrina, and Glycine (soybean). The extent of acetylene reduction varied depending on both the Rhizobium strain and O2 tension, suggesting that reduction activity differs according to the cellular organization of individual strains.

Two strains of bacteria regarded as belonging to a new species were isolated from bodies of water near Baton Rouge, La. The cells of these strains were gram-negative, curved rods, the degree of curvature varying among cells in a single culture. A pink pigment was produced on glucose-peptone-yeast extract agar. The strains were nonmotile and nonfermentative, and the guanine-plus-cytosine contents of their deoxyribonucleic acids varied from 49.3 to 49.6 mol%. The species cannot be assigned to any known genus, and therefore a new genus, Runella, is proposed, with R. slithyformis as the type species. The type strain of this species is strain 4 (= ATCC 29530). At present, it is difficult to place the genus Runella in a family.

Cultures of the redmouth (RM) bacterium, one of the etiological agents of redmouth disease in rainbow trout (Salmo gairdneri) and certain other fishes, were characterized by means of their biochemical reactions, by deoxyribonucleic acid (DNA) hybridization, and by determination of guanine-plus-cytosine (G+C) ratios in DNA. The DNA relatedness studies confirmed the fact that the RM bacteria are members of the family Enterobacteriaceae and that they comprise a single species that is not closely related to any other species of Enterobacteriaceae. They are about 30% related to species of both Serratia and Yersinia. A comparison of the biochemical reactions of RM bacteria and serratiae indicated that there are many differences between these organisms and that biochemically the RM bacteria are most closely related to yersiniae. The G+C ratios of RM bacteria were approximated to be between 47.5 and 48.5%. These values are similar to those of yersiniae but markedly different from those of serratiae. On the basis of their biochemical reactions and their G+C ratios, the RM bacteria are considered to be a new species of Yersinia, for which the name Yersinia ruckeri is proposed. Strain 2396-61 (= ATCC 29473) is designated the type strain of the species.

A Streptomyces strain, AUW-83, isolated as a contaminant in a marine fungal culture and previously reported to produce the chromomycin antibiotics, was taxonomically evaluated by direct comparison with strains selected from those species which were previously studied and reported by collaborators of the International Streptomyces Project and with additional strains. Strain AUW-83 was found to resemble strain 829 of the flavensomycin-producing species, Streptomyces cavourensis Giolitti 1958 (which name was not effectively published, according to Rule 25b of the Bacteriological Code, 1976 revision), more closely than strain 689 of the xanthicin-producing species, S. xanthochromogenus (sic) Arishima, Sakamoto, and Sato 1956. Two additional strains of the “griseus” series—Illinois 205-2 and Illinois 205-2M—reputed to produce flavensomycin were observed to be phenotypically very similar both to strain 829 and to strain AUW-83. These four strains appear to belong to one and the same species, for which we propose the name Streptomyces cavourensis. Because the name S. cavourensis was not effectively published by Giolitti, it is available for use (as a revived name [nom. rev.]) for the same or for a different species. To avoid confusion and chaos, we have elected to use the name S. cavourensis for the same organism to which Giolitti originally applied the name. However, under the circumstances, the name S. cavourensis is to be attributed to the present authors, namely Skarbek and Brady, not to Giolitti. The production of structurally unrelated antibiotics is regarded by us as a major indication for subspecific differentiation of strain AUW-83 from the flavensomycin-producing strains of S. cavourensis, and so we place strain AUW-83 in a separate subspecies, for which we propose the name Streptomyces cavourensis subsp. washingtonensis subsp. nov. Strain 829 of Giolitti (= ATCC 14889 = CBS 669.69 = NRRL 2740) is designated the type strain of S. cavourensis, and strain AUW-83 (= ATCC 27732 = NRRL B-8030) is designated the type strain of S. cavourensis subsp. washingtonensis. The valid publication of this subspecies name automatically creates the publication of the name of the type subspecies—S. cavourensis subsp. cavourensis Skarbek and Brady—the nomenclatural type of which is strain 829.

Fifteen strains of Actinomycetales alleged to produce antibiotics belonging to the aureolic acid group, some of which are useful as antitumor agents, were compared taxonomically with Streptomyces cavourensis subsp. washingtonensis strain AUW-83, a strain which produces chromomycin antibiotics. Of these strains, Streptomyces aburaviensis ATCC 23869, Streptomyces minutiscleroticus ATCC 17757, and S. cavourensis subsp. washingtonensis strain AUW-83 are type strains. None of the strains were regarded as similar enough to strain AUW-83 to be placed in S. cavourensis subsp. washingtonensis. Bristol’s Streptomyces sp. C-14795, a chromomycin-producing strain which has not been previously reported in the literature, and S. aburaviensis ATCC 23869 appear to be different strains of the same species; the same is true of Streptomyces griseus no. 7 and Streptomyces sp. KCC S-1081 and of Streptomyces argillaceus F.D. no. 6590 and Streptomyces sp. M-198. However, to establish the identities and taxonomic relationships of these 15 strains, further studies involving direct comparisons with type and/or neotype strains of closely related nomenspecies will be required.

A patient under immunosuppressive treatment of Hodgkin’s disease developed generalized skin granulomata and subcutaneous abscesses. Several aspirated pus samples yielded acid-fast rods with the following properties: Temperature optimum, about 30°C with no growth at 37°C; slow growth (2 to 4 weeks); nonchromogenic; hemoglobin or hemin requirement for growth; catalase negative; pyrazinamidase and nicotinamidase positive; and urease negative. The guanine-plus-cytosine content of the deoxyribonucleic acid was calculated from the melting temperature to be 66.0 mol%. It is concluded that these isolates belong to a new species, for which the name Mycobacterium haemophilum is proposed. The type strain of this species is strain 1 (= ATCC 29548). The new species is related to M. marinum and M. ulcerans.

Six Acholeplasma strains, C1, C112, C117, C581, C589, and PC536, isolated from the lungs, and in one case also from the liver, of aborted horse fetuses exhibited serological properties distinguishing them from the known Acholeplasma species. From the results obtained in growth inhibition, metabolic-inhibition, indirect immunofluorescence, and double-immunodiffusion tests, it can be concluded that these strains belong to two new species, for which the names Acholeplasma equifetale and Acholeplasma hippikon are proposed. A. equifetale contains strains C112, C117, C581, and C589, of which C112 (= ATCC 29724) is the type strain. A. hippikon contains strain C1 and PC536, of which C1 (= ATCC 29725) is the type strain.

Five new strains of yellow-pigmented, gram-negative, motile, hydrogen-oxidizing bacteria were isolated; each served as a host for simultaneously isolated bacteriophages. These isolates and two additional strains were compared with other gram-negative, hydrogen-oxidizing bacteria with respect to morphology; nutritional and biochemical properties; growth parameters; cytochrome content; pigment production; susceptibility to bacteriophages, bacteriostatic agents, and antibiotics; deoxyribonucleic acid base composition; and deoxyribonucleic acid-deoxyribonucleic acid homology. Six of the strains were characterized by a high degree of interstrain similarity and were found to be related to Pseudomonas flava. However, due to basic differences between these strains and P. flava, the former are regarded as comprising a new species for which, because of its moderate relationship to P. flava, the name Pseudomonas pseudoflava is proposed. The type strain of P. pseudoflava, GA3, has been deposited with the Deutsche Sammlung von Mikroorganismen in Göttingen under the number DSM 1034.

The suitability of the growth precipitation test for differentiation of 43 Acholeplasma strains belonging to seven different species was examined. Cross-growth precipitation tests indicated a high degree of specificity of this method, which makes possible the simple and reliable identification of Acholeplasma species.

A numerical taxonomic study was carried out to establish: (i) the relationship of freshly isolated, oxidase-negative, vibrio-like organisms to strains listed in certain culture collections as Vibrio metschnikovii and V. proteus; (ii) the relationship of all these oxidase-negative organisms to the genus Vibrio and related organisms. Eighty-three strains were tested for their abilities to grow on 49 compounds as the sole organic source of carbon, their fermentation of carbohydrates, and their production of extracellular enzymes; various physiological tests were also carried out. In all, 130 characters were determined for each strain. The collection of strains were of Vibrio and Aeromonas species, certain other possibly related organisms, and 40 oxidase-negative organisms from a wide range of habitats. It was concluded that all of the oxidase-negative, vibrio-like strains except one belong to the species Vibrio metschnikovii Gamaleia 1888 (synonyms V. proteus Buchner 1885; V. cholerae biotype proteus Shewan and Veron 1974). The lectotype is NCTC 8443.

A facultatively autotrophic, hydrogen-oxidizing spirillum was isolated from a small eutrophic lake in Switzerland. It is medium-sized (0.6 to 0.8 μm) and possesses bipolar tufts of flagella. The guanine-plus-cytosine content of its deoxyribonucleic acid is 60 to 62 mol%. Based upon its source, morphology, and biochemical features, the organism belongs to the genus Aquaspirillum Hylemon et al. 1973. Because this organism does not closely resemble any of the previously described medium-sized aquaspirilla, it is proposed here as belonging to a new species, Aquaspirillum autotrophicum. The specific epithet refers to the outstanding property of this organism to grow on hydrogen and carbon dioxide as energy and carbon sources, respectively. The type strain of A. autotrophicum, strain SA 32, was deposited with the Deutsche Sammlung von Mikroorganismen under the number DSM 732.

Ten nonfluorescent Pseudomonas strains isolated from water-soaked lesions on cotyledons of plants of five Citrullus lanatus (watermelon) plant introductions were characterized and compared phenotypically with 22 other pseudomonads. The strains were distinguished phenotypically from other known plant pathogenic pseudomonads. The watermelon bacterium was aerobic. Cells were rod-shaped, gram negative, and motile by means of a single polar flagellum. They were nonfluorescent and grew at 41°C but not at 4°C. Oxidase production and the 2-ketogluconate reaction were positive. The 10 strains utilized β-alanine, L-leucine, D-serine, n-propanal, ethanol, ethanolamine, citrate, and fructose for growth. No growth occurred with sucrose or glucose. Their deoxyribonucleic acid base composition was 66 ± 1 mol% guanine plus cytosine. The bacterium is phenotypically similar to P. pseudoalcaligenes but differs from it in being pathogenic to watermelon, Cucumis melo (cantaloupe), Cucumis satiwus (cucumber), and Cucurbita pepo (squash). The name P. pseudoalcaligenes subsp. citrulli is proposed for the new subspecies, of which strain C-42 (= ATCC 29625) is the type strain.

The role of opportunistic pathogens in infectious pathological conditions of humans is increasingly being appreciated. Some organisms that have been considered nonpathogenic occasionally cause severe hospital infections and outbreaks of acute enteric diseases.

Chloropseudomonas ethylica 2K cultured at Brookhaven National Laboratory since 1962 has always been a mixed culture of Prosthecochloris aestuarii and Desulfuromonas acetoxidans.

The characteristics of ATCC 11337, the type strain of Spirillum lunatum Williams and Rittenberg 1957, do not fit the original description of the species. Cultures of this same strain held by the National Collection of Marine Bacteria under the number 54 appear to consist of two kinds of organisms: (i) relatively small, short, vibrio-shaped rods having single flagella, capable of growing in the presence or absence of seawater and of catabolizing certain sugars, incapable of forming coccoid bodies, and having a deoxyribonucleic acid base composition of 63 to 64 mol% guanine plus cytosine (these organisms closely resemble those found in cultures of ATCC 11337), and (ii) relatively large organisms having a typical spirillum morphology with bipolar tufts of flagella, requiring seawater for growth, capable of forming coccoid bodies, incapable of catabolizing sugars, and having a deoxyribonucleic acid base composition of 45 mol% guanine plus cytosine. The larger organisms have been placed in a new subspecies of Oceanospirillum maris, O. maris subsp. williamsae, the type strain of which is ATCC 29547. The smaller organisms do not appear to belong to either the genus Oceanospirillum or the genus Aquaspirillum, and their classification is not yet apparent. Because the name Spirillum lunatum has been a source of confusion, it is requested that the Judicial Commission place this name on the list of rejected names as a nomen confusum.