- Volume 30, Issue 2, 1980

University of Alberta Soil Microbiology strain 9D is a gram-negative, nonflagellated, gliding, white, facultatively anaerobic, non-polysaccharolytic, long, thin rod or filament. The guanine plus cytosine content of the deoxyribonucleic acid of this strain is 37.0 mol©. This strain belongs to the genus Flexibacter, but it differs significantly from strains of previously described species. Therefore, it is regarded as belonging to a new species, for which the name Flexibacter canadensis is proposed. A culture of the type strain, 9D, has been deposited in the American Type Culture Collection under the number 29591.

A convenient method has been devised to chromatograph simultaneously numerous samples of reassociated deoxyribonucleic acid under thermoregulated conditions. Small amounts of hydroxyapatite (0.2-ml bed) were used, and the effluent volumes for separating single-stranded deoxyribonucleic acid from deoxyribonucleic acid duplexes were 1.5 ml each. The method was comparable in reliability and reproducibility to other well-established techniques, but it combined improvements in ease of operation, in rapidity, and in simplicity, which are desirable in large taxonomic studies.

Nine strains of Pseudomonas maltophilia which do not require methionine are described. Methionine-requiring and -nonrequiring strains of P. maltophilia belong to different biovars, which are designated I and II, respectively; they differ from each other with respect to phenotypic characteristics, cellular fatty acid composition, deoxyribonucleic acid base composition, and deoxyribonucleic acid-deoxyribonucleic acid hybridization. The relationship of P. maltophilia to Xan-thomonas species is also discussed.

Eight strains each of Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, and Bacillus amyloliquefaciens were analyzed by using pyrolysis gas-liquid chromatography. Statistical analysis with canonical variates gave four well-separated groups, which represented the four species. Further analysis of the same strains by deoxyribonucleic acid-deoxyribonucleic acid hybridization and API identification systems confirmed the discrimination obtained with pyrolysis gasliquid chromatography. However, analysis by biochemical tests performed in the classical way gave only three groups since it was not possible to achieve separation of the strains representing B. subtilis from those of B. amyloliquefaciens when these tests were used.

When polyacrylamide gel electrophoretic profiles of soluble proteins of seven Arthrobacter strains were compared with those of seven coryneform and coryneform-like species, the arthrobacters were delineated from the latter species. Comparisons were made from a composite gel by calculating similarity coefficients. When Arthrobacter globiformis was used as a reference, similarity coefficients for the other Arthrobacter species ranged from 20.7 to 34.5, but the similarity coefficients for the other genera tested were only 7.9 to 18.5. In addition to a separation of arthrobacters from other coryneforms by this method, Arthrobacter crystallopoites and Arthrobacter pyridinolis had a similarity coefficient of approximately 95, which set these two apart from the other Arthrobacter species studied and furthermore suggested that they represent one species. Gel electrophoresis of soluble proteins may provide a good alternative to the use of rod to coccoid transition for identifying arthrobacters since it is faster and simpler to perform than chemical analyses of cell wall components.

A total of 124 strains representing Corynebacterium diphtheriae (mitis, gravis, intermedius, and ulcerans) and 12 strains of the closely related Corynebacterium pseudotuberculosis (Corynebacterium ovis) were found to be pyrazinamidase negative (Pyz-). By contrast, a large number of strains representing various corynebacterial species, nocardias, bacterionemas, actinomyces, and rothias were pyrazinamidase positive (Pyz+). The usefulness and possible implications of this finding are discussed, and a rapid test for pyrazinamidase activity is described.

Flavobacterium heparinum was inadequately characterized when it was originally described by Payza and Korn in 1956. As a result it was listed as a species incertae sedis in the 8th edition of Bergey’s Manual. The authors isolated a single strain of this organism, and presumably it was this strain which was deposited in the American Type Culture Collection (ATCC) under the number 13125 (= National Collection of Industrial Bacteria [NCIB] 9290). Because the original description was based on a single strain, ATCC 13125 is here regarded as the type strain of F. heparinum by monotypy. The results of an extensive study of the morphological, physiological, biochemical, and antimicrobial (lytic) properties of this strain suggest that it is more properly classified in the genus Cytophaga as C. heparina (Payza and Korn) comb. nov.

The genetic relatedness of 62 strains of Bacillus sphaericus, 4 strains of B. pasteurii, 6 strains of B. brevis, and 1 strain each of B. globisporus and B. aminovorans was examined by deoxyribonucleic acid (DNA) homology. Five homology groups were identified, and they contained all but 12 of the B. sphaericus strains. Intergroup homologies ranged from about 20 to 40© except for groups III and IV, which had 50 to 60© intergroup homology. Although a few strains within some groups had 63 to 69© homology to the reference strain, most strains were in the 70 to 90© range. Homology group I contained the type of strain of B. sphaericus, ATCC 14577, and three other strains. Homology group II was subdivided into groups IIA and IIB, the former containing all of the strains found to be pathogenic for mosquito larvae. Although none of the physiological characteristics or combination of characteristics examined allowed the complete differentiation of strains into the appropriate homology groups, the strains of group I could be distinguished from those of group IIA-IIB. The strains of group IIA could be distinguished from all others by mosquito pathogenicity. The B. pasteurii strains formed a distinct homology group, although the guanine plus cystosine contents of their DNAs fell within the range of 34 to 37 mol© found for the 45 strains of B. sphaericus examined for this property. The B. brevis strains possessed a higher guanine plus cystosine content, and their DNAs were unrelated to the DNAs of the B. sphaericus reference strains tested.

The morphological, biochemical, and physiological characteristics of nine bacterial strains isolated from a solar salt facility located on Bonaire, Netherlands Antilles are described. The bacteria were gram-negative rods which produce white, opaque colonies on solid media. During the log phase of growth, the cultures consisted of single and paired cells with polar flagella predominating. Older cultures characteristically produced highly elongated, flexible rods. All of these strains reduced NO3 to NO2, grew anaerobically in the presence of NO3, and fermented glucose but oxidized sucrose, glycerol, mannose, and cellobiose. All strains were ornithine and lysine decarboxylase positive, catalase positive, and cytochrome oxidase negative. Eight of the nine strains grew in a complex Casamino Acids liquid medium containing from 0 to 32© (wt/vol) solar salt at temperatures from 23 to 37°C; the ninth strain was restricted in its growth to 0 to 20© solar salt. The guanine plus cytosine content of the deoxyribonucleic acid was 61 ± 1 mol©. This combination of morphology, salt tolerance, and guanine plus cytosine content supports the establishment of a new genus, Halomonas, in Family II (Vibrionaceae) of part 8, Gram-Negative Facultatively Anaerobic Bacteria, of Bergey’s Manual (8th edition). The type species of this genus is H. elongata, the type strain of which is isolate 1H9 (= ATCC 33173). Strain 1H15 is regarded as belonging to a biovar of H. elongata on the basis of its production of lophotrichous cells and its inability to grow at 37°c in the presence of 32© solar salt.

Isolates of the kidney disease bacterium, the causative agent of bacterial kidney disease in salmonid fishes, were characterized by the guanine plus cytosine content of the deoxyribonucleic acid, by cell wall sugar composition, and by amino acid composition of the peptidoglycan cell wall layer. The guanine plus cytosine contents of the deoxyribonucleic acids of the isolates were 53 ± 0.46 mol©. Glucose was the principal cell wall sugar detected in each kidney disease bacterium isolate. The major amino acids detected were alanine, glutamic acid, lysine, and glycine. It is proposed that these organisms form a single species belonging to a new genus, for which the name Renibacterium is proposed. The type species is Renibacterium, salmoninarum sp. nov., of which the type strain is Lea-1-74 (= ATCC 33209). This organism is most closely related to the Coryneform Group of Bacteria.

In the course of a taxonomic study of methanol-assimilating yeasts, a number of isolates were obtained from various natural habitats in Japan, and from these three new species, Pichia cellobiosa, Candida cariosilignicola, and Candida succiphila, have been described. P. cellobiosa differs from the related species Pichia castillae with respect to fermentation and assimilation of sugars, biotin requirement for growth, and lower maximum temperature for growth. C. cariosilignicola differs from the related species Torulopsis apicola and Torulopsis bombicola with respect to assimilation of sugars and proton magnetic resonance spectra of cell wall mannans. C. succiphila is similar to C. cariosilignicola but differs from it with respect to fermentation and assimilation of sugars, lower maximum temperature for growth, higher deoxyribonucleic acid base composition, and proton magnetic resonance spectra of cell wall mannans. The type strains of these new species are KL 1 (=IAM 12481), KL 24 (=IAM 12484), and KL 30 (=IAM 12489), respectively. Cultures of the type strains have been deposited in the Institute of Applied Microbiology Culture Collection Center, University of Tokyo, Tokyo, Japan, and the Institute for Fermentation, Osaka, Japan.