- Volume 43, Issue 4, 1993

Haemophilus influenzae is a V factor-dependent species. A plasmid conferring V factor independence in Haemophilus parainfluenzae and Haemophilus ducreyi was transferred to plasmid-free H. influenzae Rd by DNA transformation. The growth characteristics of the transformants in a complex and a chemically defined medium were compared, and the ability of several exogenous pyridine nucleotides and precursors to support growth was examined. Although the transformants appeared to be V factor independent in a complex medium, in a chemically defined medium they exhibited both V factor-dependent and nicotinamide-dependent growth. Because of the inability of the plasmid-free H. influenzae Rd to utilize nicotinamide for growth, it was concluded that the genes conferring this function were plasmid linked. Our results indicate that the V factor requirement, as it is presently defined, is not suitable to serve as a definitive taxonomic criterion for species determination in the family Pasteurellaceae.

Thirteen strains of motile arthrospore-bearing actinomycetes were isolated from soils in Japan, India, and Nepal. These organisms underwent differentiation into aerial and substrate mycelia in their growth cycles and produced motile spores arranged in chains on their aerial mycelia. Sporangia were not observed. The cell walls contained L-lysine, serine, glycine, D-glutamic acid, and D-alanine (cell wall type VI). Mycolic acids were absent; the major menaquinones were MK-9(H8) and MK-10(H8); the phospholipid type was type PIII. The taxonomic characteristics of these isolates are different from those of the previously described motile actinomycetes. The name Catenuloplanes japonicus gen. nov., sp. nov., nom. rev. is proposed for these organisms. The type strain is strain N381-16, which has been deposited in the Institute for Fermentation, Osaka as strain IFO 14176 and in the American Type Culture Collection as strain ATCC 31637.

The phosphatase activities of 114 micrococcal strains belonging to seven different species and of an additional 150 unspeciated micrococcal strains were evaluated on solid media at various pHs containing or not containing phosphates. In the presence of phosphates, only nine strains (five unspeciated strains, one Micrococcus luteus strain, and three Micrococcus varians strains) yielded a positive reaction on plates at pH 8. In media (at pH 8) deprived of phosphates, in contrast, all but 15 strains demonstrated clear-cut phosphatase activity. Acid phosphatase could not be evaluated on solid media since none of the strains grew satisfactorily on plates at pH 5. The phosphatase activities of seven (one or two for each species, which included phosphatase-negative strains) of the strains whose colonies proved phosphatase negative at pH 8 and of 18 (two or three strains per species) of those with phosphatase-positive colonies were evaluated at pH 5 and 8.5 in toluene-treated cells which had been grown in liquid media at pH 7 containing or not containing phosphates. All strains demonstrated distinct phosphatase activity at both pHs when grown in media not containing phosphates. In contrast, when strains were grown in the presence of such substances, virtually no activity was observed at pH 8.5, and, generally, a much reduced activity was observed at pH 5. The phosphatase activity of micrococci of the various species (three to eight strains per species) was also compared with that of staphylococci of different species (5 to 10 strains per species) by the methyl green-phenolphthalein diphosphate method, the sensitivity of which can be varied by using different enzyme substrates. By using phenolphthalein diphosphate as an enzyme substrate, it was found that virtually all the different species of staphylococci yielded a positive reaction on plates not containing phosphates while almost all micrococci proved phosphatase negative with the methyl green-phenolphthalein diphosphate method. This indicates that the phosphatase activity of micrococci can easily be differentiated from that of staphylococci.

DNA-DNA hybridization studies (with the S1 nuclease method [J. L. Johnson, Methods Microbiol. 18:33-74, 1985]) were performed on members of the genus Fusobacterium. Fusobacterium varium ATCC 8501Tshowed 88 and 79% DNA homology with F. pseudonecrophorum JCM 3722Tand JCM 3723, respectively, while F. pseudonecrophorum JCM 3722Tshowed 81 and 82% DNA homology with F. varium ATCC 8501Tand F. pseudonecrophorum JCM 3723, respectively. These genetic data and their similar phenotypic characteristics suggest that F. varium(Eggerth and Gagnon 1933) Moore and Holdeman 1969 and F. pseudonecrophorum(ex Prévot 1940) Shinjo et al. 1990 belong to a single species. We propose, therefore, that strains JCM 3722 and JCM 3723 of F. pseudonecrophorum be transferred to the species F. varium.

DNA relatedness was evaluated among the type strains of Streptomyces lavendulae and the 11 species which were designated as subjective synonyms in volume 4 of Bergey’s Manual of Systematic Bacteriology. Three S. lavendulae subspecies, five putative strains of S. lavendulae, and the type strain of “S. majorciensis” were also included in the present study. The 21 strains studied segregated into 14 clusters (average linkage) when grouped at >70% DNA relatedness, including 10 single-member clusters. S. lavendulae, S. lavendulae subsp. avirens, S. lavendulae subsp. grasserius, and S. columbiensis were found to be synonymous at >79% DNA relatedness. The other clusters, on the basis of their relatively low DNA relatedness to S. lavendulae, appear to be valid species rather than subjective synonyms of S. lavendulae.

Phylogenetic relationships among strains of Zoogloea and related taxa were determined by 16S rDNA sequencing and genomic DNA hybridization techniques. The 16S rRNA gene was amplified by the polymerase chain reaction with a pair of eubacterial consensus primers and sequenced directly by using an automated fluorescent DNA sequencer. Sequence comparisons and distance matrix tree analysis revealed that Zoogloea ramigera IAM 12136 (= N. C. Dondero 106, type strain) and Zoogloea sp. ATCC 19324 formed a lineage with Rhodocyclus purpureus in the β subclass of Proteobacteria. Z. ramigera IAM 12670 (= P. R. Dugan 115) was shown to belong to another cluster with Alcaligenes eutrophus and Pseudomonas cepacia in the β subclass. In contrast, Z. ramigera IAM 12669 (= K. Crabtree I-16-M) proved to be a member of the ± subclass of the Proteobacteria, closely related to Agrobacterium tumefaciens. Genomic DNA hybridization studies also showed that there is genetic diversity among the strains currently designated Z. ramigera, but typical Zoogloea strains, characterized by their production of rhodoquinones, are highly related to each other and can be regarded as a single species. On the basis of the molecular data, together with the early phenotypic and chemotaxonomic information, we have emended the generic description of Zoogloea.

Partial 16S rRNA gene sequences (16S rDNA) of Gemella morbillorum and Gemella haemolysans were determined by sequencing polymerase chain reaction-amplified DNA. A phylogenetic analysis of 53 eubacterial 16S rRNA sequences grouped the gemellae on a distinct branch separate from the 18 members of the genus Streptococcus. DNA-DNA hybridization results indicate that the two gemellae are related at the genus level but are not a single species.

We propose the name Rickettsia massiliae sp. nov. (type strain, Mtul in the Collection of the World Health Organization Collaborative Center for Rickettsial Reference, Marseille, France) for a spotted fever group rickettsia determined to be distinct from previously recognized species by the serotyping method (L. Beati, J.-P. Finidori, B. Gilot, and D. Raoult, J. Clin. Microbiol. 30:1922-1930, 1992). This rickettsia has biological characteristics similar to those of the other spotted fever group rickettsiae. In addition, a sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein analysis, a polymerase chain reaction followed by a restriction fragment length polymorphism analysis of DNA fragments, and pulsed-field electrophoresis of the genome of R. massiliae revealed unique migration patterns distinct from those of all previously recognized spotted fever group rickettsiae. These additional characteristics (Beati et al., J. Clin. Microbiol. 30:1922-1930, 1992), together with the data usually considered sufficient for description of rickettsiae, are crucial to the proposal of this new species and should be helpful in species identification.

Strains of a suggested novel type of mycobacterium have been repeatedly isolated from patients with AIDS. We summarize the results of tests performed to determine enzymatic activities and metabolic properties, the results of fatty acid analyses, and the results of a comparative 16S rRNA sequence determination. We propose formally that this organism represents a new species, Mycobacterium genavense. The type strain is strain 2289, a culture of which has been deposited in the American Type Culture Collection as strain ATCC 51234.

The base compositions of DNAs from nine Microcystis strains, as determined by high-performance liquid chromatography, were 41 to 42 mol% G+C. Chromosomal DNAs derived from these strains were found to be extremely resistant to many restriction endonucleases, and a restriction analysis revealed the presence of a dam-like methylase or both dam- and dcm-like methylases in all of the strains examined. Genomic Southern hybridization in which a synthetic oligonucleotide probe (rpoD probe) was used showed that members of the genus Microcystis might have multiple rpoD homologs, and the hybridization signal patterns observed with the DNAs of Microcystis aeruginosa strains were different from each other.

The description of Corynebacterium striatum(Chester 1901) Eberson 1918ALin Bergey's Manual of Systematic Bacteriology has many inconsistencies with the identification scheme from the Centers for Disease Control and Prevention. We have studied the four C. striatum reference strains available from the American Type Culture Collection and the National Collection of Type Cultures and found that they fit the Centers for Disease Control and Prevention description of this species. However, it appears that the wrong strains were deposited for this species, because none of the reference strains fits the descriptions in the original publications. This is a substantial case for declaring it a nomen dubium, but the name could be rescued if a careful search reveals a strain that was used in making the original description. It is hoped that some readers may have the missing strains labeled with the early names Bacillus striatus, Bacillus flavidus, or Corynebacterium flavidum.

We clarified the taxonomic position of Pseudomonas perfectomarina ZoBell in the species Pseudomonas stutzeri. Both genotypic and phenotypic observations led us to conclude that this strain belongs to genomovar 2 of P. stutzeri, for which strain ATCC 17591 is the reference strain. The uncertainty about the motility of strain ZoBell was also clarified. We found that strain ZoBell is motile by means of one polarly inserted flagellum.

The 16S rRNA gene sequences of the type strains of Aeromonas enteropelogenes and Aeromonas ichthiosmia were determined by polymerase chain reaction direct sequencing in order to clarify their interrelationships with other aeromonad species. On the basis of 16S rRNA gene sequence analysis, A. enteropelogenes and A. ichthiosmia were found to be identical to Aeromonas trota and Aeromonas veronii, respectively.

Phylogenetic and phenotypic evidence demonstrates the taxonomic heterogeneity of the genus Thermobacteroides and indicates a close relationship between Thermobacteroides acetoethylicus and members of the genus Thermoanaerobacter. Since T. acetoethylicus is the type species of Thermobacteroides, its removal invalidates the genus. As a consequence, the remaining species Thermobacteroides proteolyticus is proposed as the type species of the new genus Coprothermobacter gen. nov., as Coprothermobacter proteolyticus comb. nov.