- Volume 38, Issue 4, 1988

A total of 39 strains assigned to the strictly anaerobic species Butyrivibrio fibrisolvens were examined for deoxyribonucleic acid (DNA) relatedness. The guanine-plus-cytosine (G+C) base contents determined from the buoyant densities of chromosomal DNAs of 27 strains were 39.0 to 42.0 mol%. Another nine strains had G+C contents of 42.0 to 45.0 mol%, and three other strains had higher G+C contents (46.4 to 49.2 mol%). Genetic relationships among the strains were determined by DNA hybridizations, using both spectrophotometric and membrane filter techniques. The relationships derived by both methods are compared. The G+C content and hybridization results indicate that the strains comprise a genetically heterogeneous group, representing a number of distinct species. Strain D1, the type strain of B. fibrisolvens, was not closely related to any of the other 38 strains. Five groups of related bacteria, encompassing 20 Butyrivibrio strains, were identified as forming five separate species. The largest group identified by DNA relatedness as belonging to the same species contained only six strains. The other 19 strains were not closely related to any other strain.

Seventeen strains corresponding to Flavobacterium spiritivorum, seven strains corresponding to or received as Sphingobacterium mizutae, and the type strains of Flavobacterium multivorum and Flavobacterium thalpophilum were examined for deoxyribonucleic acid-deoxyribonucleic acid relatedness. Duplicate cultures of the type strains and of some of the S. mizutae field strains, from various sources, were included to make a total of 43 cultures examined. Of the 17 F. spiritivorum strains, 15 could be included in that species, but 2 constituted a closely related, but separate, species. For this new species we propose the name Flavobacterium yabuuchiae, with strain F8081 (= NCTC 12113) as the type strain. Only three strains could be included in S. mizutae; of the four remaining strains, one represented a separate species 32% related to S. mizutae, one corresponded to Flavobacterium meningosepticum, and the classification of the other two was unresolved. The type strains of F. multivorum and F. thalpophilum were distinct from the other cultures studied. The new combination Flavobacterium mizutaii is also proposed.

The phylogenetic position of Arachnia propionica was determined by using reverse transcriptase sequencing of long regions of 16S ribosomal ribonucleic acid. Arachnia propionica formed a distinct group with Propionibacterium freudenreichii and Propionibacterium acnes but was unrelated to members of the genus Actinomyces. The retention of Arachnia as a separate genus is unjustified. On the basis of the present analysis and previous chemotaxonomic evidence, we propose that Arachnia propionica be reclassified in the genus Propionibacterium as Propionibacterium propionicus (Buchanan and Pine) comb. nov.

The fatty acid compositions and multiple antibiotic resistance patterns of 32 strains of Bradyrhizobium japonicum correlated with two major deoxyribonucleic acid homology groups. In group I, the fatty acid composition was 1.3% 16:1 cis9 acid, 3.6% 16:1C acid, 8.8% 16:0 acid, 1.2% 19:0 cyclopropane acid, and 81.2% 18:1 acid. Group II contained 0.5% 16:1C acid, 11.1% 16:0 acid, 0.8% 17:0 cyclopropane acid, 24.7% 19:0 cyclopropane acid, and 62.3% 18:1 acid. Group I strains were susceptible to rifampin (500 μg/ml), tetracycline (100 μg/ml), streptomycin (100 μ/ml), chloramphenicol (500 μg/ml), erythromycin (250 μg/ml), carbenicillin (500 μg/ml), and nalidixic acid (50 μg/ml), whereas group II strains were resistant to these antibiotics. Both groups were resistant to trimethoprim (50 μg/ml) and vancomycin (100 μg/ml).

A survey for bacterial diseases of millet and sorghum was conducted in northern Nigeria during the 1984 growing season. Bacterial diseases were prevalent throughout the area surveyed. Yellow, mucoid bacterial colonies were consistently isolated from pearl millet leaves exhibiting bacterial streak symptoms. The causative organism was characterized as a pathovar of Xanthomonas campestris. The strains isolated formed a homogeneous group of aerobic, motile, gram-negative, rod-shaped organisms which were distinctly different pathologically, serologically, and by membrane protein patterns from other pathovars of X. campestris. The cells measured 0.45 by 2.25 μm, and each cell had one polar flagellum. Optimal growth occurred between 26 and 30°C, and the maximum NaCl tolerance was 3%. Pearl millet (Pennisetum americanum) and Proso millet (Panicum miliaceum) are the only known hosts. For this pathovar, we propose the name Xanthomonas campestris pv. pennamericanum pv. nov. (the name is derived from Pennisetum americanum, the Latin binomial for pearl millet). The holopathotype strain is strain B6-P, which has been deposited in the American Type Culture Collection, Rockville, Md.

The name Campylobacter pylori subsp. mustelae subsp. nov. is proposed for a Campylobacter commonly isolated from normal or inflamed gastric mucosa of ferrets. C. pylori subsp. mustelae, like C. pylori, has multiple sheathed flagella, rapidly hydrolyzes urea, is H2S negative on triple sugar iron agar, and has a variable reaction on lead acetate strips. It does not grow in the presence of 3% NaCl, and growth is variable in 0.04% triphenyltetrazolium chloride and 1% glycine. Unlike C. pylori, this organism reduces nitrate, is susceptible to nalidixic acid, and is resistant to cephalothin. Three strains of C. pylori subsp. mustelae were highly related (≥86%) as determined by deoxyribonucleic acid (DNA)-DNA hybridization (hydroxyapatite method, 50 and 65°C). C. pylori subsp. mustelae was ≥85% related to C. pylori, whereas the level of relatedness with another seven Campylobacter isolates was ≤2% at 65°C. The type strain of C. pylori subsp. mustelae is strain R85-13-6 (= ATCC 43772), and its DNA has a guanine-plus-cytosine content of 38 mol%.

The description for three strains of Erwinia cancerogena was expanded to include 108 phenotypic characters. These strains were compared with seven strains of Erwinia carotovora subsp. carotovora and the type strains of Enterobacter (Erwinia) nimipressuralis, Enterobacter cloacae, Enterobacter amnigenus, and Enterobacter intermedium. Our results support the proposal to transfer Erwinia cancerogena to the genus Enterobacter as Enterobacter cancerogenus comb. nov.

We determined deoxyribonucleic acid (DNA) relatedness among toxigenic strains of Clostridium botulinum toxin type G and nontoxigenic strains labeled Clostridium subterminale and Clostridium hastiforme by using the hydroxyapatite method at 50 and 65°C. Seven DNA hybridization groups were detected. All of the strains were anaerobic, gram-positive, rod shaped, asaccharolytic, and proteolytic; produced acetic, butyric, isobutyric, and isovaleric acids; and were separable by additional phenotypic characteristics into the seven hybridization groups. One hybridization group was composed of all nine C. botulinum type G strains, two strains received as C. subterminale, and one strain received as C. hastiforme. These strains showed an average level of intragroup relatedness of 94% and were less than 25% related to the type strains of C. botulinum (BL 4847), C. subterminale (BL 4856), and C. hastiforme (BL 4858). The guanine-plus-cytosine contents of four strains in this group were 28 to 30 mol%. A new species, Clostridium argentinense, is proposed for the genotypically homogeneous group that is unrelated to the strains of C. botulinum and other clostridial species which we studied. The type strain of C. argentinense sp. nov. is strain ATCC 27322. Two of the six other DNA hybridization groups are represented by the type strains of C. subterminale and C. hastiforme. The four remaining DNA hybridization groups represent undescribed species that contain one or two strains each.

A new gram-positive anaerobic bacterium is described on the basis of 89 strains isolated from the intestinal tracts of red solider bugs (Pyrrhocoris apterus L.). The fermentation products of glucose are acetic acid, lactic acid, ethanol, CO2, and H2. The guanine-plus-cytosine content of the deoxyribonucleic acid is 60 to 61 mol%. The biochemical and physiological features of this organism distinquish it from members of previously described genera. We propose a new genus, Coriobacterium, and a new species, Coriobacterium glomerans. The type strain is C. glomerans PW2 (= DSM 20642).

Reverse transcriptase sequencing of long regions of 16S ribosomal ribonucleic acids of Corynebacterium paurometabolum and Rhodococcus aurantiacus was performed to determine the relationship of these organisms to other mycolic acid-containing actinomycetes. These two species were highly related to each other (more than 99% sequence homology in a large stretch of 1,184 nucleotides) but were distinct from the genera Corynebacterium, Mycobacterium, Nocardia, and Rhodococcus. On the basis of our findings and previous findings we propose that C. paurometabolum and R. aurantiacus be reduced to a single species and reclassified in a new genus, Tsukamurella. The type species is Tsukamurella paurometabolum comb. nov.

A total of 33 strains of fast-growing soybean rhizobia isolated from soil and soybean nodules collected in China and 25 strains belonging to the genera Rhizobium, Bradyrhizobium, and Agrobacterium were compared by numerical taxonomic techniques, using 240 different characters. Our results indicated that all of the strains of fast-growing soybean rhizobia which we examined are closely related (guanine-plus-cytosine content, 59.9 to 63.8 mol%) and are separated from Rhizobium and Bradyrhizobium at the generic level. Based on numerical taxonomy, deoxyribonucleic acid (DNA) base ratio determinations, DNA-DNA hybridization data, serological analysis data, the composition of extracellular gum, bacteriophage typing data, and soluble protein patterns, we propose that the fast-growing soybean rhizobia represent members of a new genus rather than a species of Rhizobium (Rhizobium fredii); we propose Sinorhizobium gen. nov. as an appropriate generic name. The type species of the new genus is Sinorhizobium fredii comb. nov. (basonym, Rhizobium fredii Scholla and Elkan 1984), and the type strain is strain ATCC 35423 (= USDA 205). For the other species we propose the name Sinorhizobium xinjiangensis sp. nov.; the type strain of this species is strain CCBAU 110, which has been deposited in the Beijing Agricultural University Culture Collection, Beijing, People’s Republic of China.

Representatives of Saccharomonospora viridis (type strain DSM 43017), Saccharomonospora caesia (type strain DSM 43044), and Saccharomonospora internatus (type strain DSM 43671), together with 52 Saccharomonospora isolates from compost, manure, hay, and soil, were characterized by determining morphological, biochemical, and physiological properties, phage sensitivity, antibiotic activity, enzyme and protein patterns (polyacrylamide gel electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis), and restriction patterns of chromosomal deoxyribonucleic acids. The type strain of S. internatus proved to be identical to S. viridis, and thus S. internatus should be regarded as a synonym of S. viridis. A total of 16 strains differed significantly from S. viridis and S. caesia and are proposed as members of a new species, Saccharomonospora glauca (type strain, DSM 43769). Since the three species differ in phage sensitivity, production of antibiotic substances, and enzyme, protein, and deoxyribonucleic acid restriction patterns, these properties can be used for reliable species identification.

In this study we examined the taxonomic relationships of strains variously labeled Achromobacter species biotypes 1 and 2, Achromobacter group A, and Centers for Disease Control (CDC) groups Vd-1 and Vd-2. Previous studies of ribosomal ribonucleic acid cistron similarities placed these organisms on the Brucella ribosomal ribonucleic acid branch of ribosomal ribonucleic acid superfamily IV; their closest neighbors were Brucella, Phyllobacterium, and the Agrobacterium-Rhizobium complex. We performed a numerical taxonomic analysis of 284 phenotypic features (69 conventional tests, 147 API assimilation tests, 68 API ZYM tests) carried out on 95 strains. These organisms comprised 56 strains thought to correspond to CDC group Vd (including 3 strains originally labeled “Pseudomonas arsenoxydans”) and 39 strains (included for reference purposes) representing the genera Achromobacter, Agrobacterium, Alcaligenes, Brucella, Mycoplana, Phyllobacterium, and Rhizobium. A phenotypic analysis showed that group Vd bacteria are most similar to Phyllobacterium. However, strains of Group Vd were shown to be distinct by deoxyribonucleic acid (DNA)-DNA hybridization and by several phenotypic tests from Phyllobacterium and other related taxa. The CDC group Vd strains formed essentially a single taxon in the numerical taxonomic analysis of phenotypic characters and as determined by DNA-DNA hybridization. This taxon could be subdivided into three biotypes (biotypes A, C, and D), but none of these corresponded to the two biotypes originally described among the group Vd strains. For CDC group Vd we propose a new genus and new species, Ochrobactrum anthropi: The type strain is strain CIP 82.115 (= CIP 14970 = NCTC 12168 = LMG 3331). O. anthropi strains are rod shaped, aerobic, gram negative, nonpigmented, and motile by means of peritrichous flagella, produce acid from several carbohydrates, and reduce both nitrate and nitrite. The guanine-plus-cytosine contents of the DNAs of 29 strains ranged from 56 to 59 mol%. Almost all 56 group Vd strains were originally isolated from various human clinical specimens, commonly from blood cultures.

We studied the purine enzyme activities in dialyzed cytoplasmic extracts from the following eight species, representing four genera, of Mollicutes: Mycoplasma pneumoniae FHT (T = type strain) and M129, Mycoplasma bovigenitalium PG-11T, Mycoplasma hominis PG-21T and 1620, Mycoplasma genitalium G-37T, Mycoplasma hyopneumoniae JT, Ureaplasma urealyticum T960T, Spiroplasma citri Maroc-R8A2T, and Anaeroplasma intermedium 5LA. In an investigation of purine nucleoside kinase activity we also included M. hominis 13408, 10144, 13428, 1612, 1184, and Botte. All of these Mollicutes species except U. urealyticum had purine phosphoribosyltransferase activity for adenine, hypoxanthine, and guanine; U. urealyticum had only adenine phosphoribosyltransferase activity. All of the organisms had nucleoside phosphorylase activity which used either ribose 1-phosphate or deoxyribose 1-phosphate and adenine, hypoxanthine, or guanine for the synthesis of nucleosides and adenosine, deoxyadenosine, guanosine, deoxyguanosine, inosine, or deoxyinosine in the reverse direction. All had 5′-nucleotidase activity for adenosine monophosphate, deoxyadenosine monophosphate, inosine monophosphate, or guanosine monophosphate. Only M. hominis 1620, 13408, 10144, and 13428, A. intermedium, and S. citri had pyrophosphate-dependent nucleoside kinase activity. Only S. citri had nucleoside kinase activity with adenosine triphosphate and deoxyguanosine. We studied pyrimidine enzyme activities in all of the Mollicutes species except M. hominis and M. bovigenitalium. All of the Mollicutes species assayed had thymidine, thymidylate, and deoxycytidine kinase and thymidine and uridine phosphorylase activities. All of the Mycoplasma spp. had deoxycytotidine monophosphate and cytidine-deoxycytidine deaminase activities. All of the Mycoplasma spp. and U. urealyticum lacked deoxyuridine triphosphatase activity. U. urealyticum lacked deoxycytidine monophosphate deaminase activity, but otherwise it resembled all of the Mycoplasma spp. A. intermedium and S. citri differed from each other and from Mycoplasma spp. and U. urealyticum in the patterns of pyrimidine enzyme activities. Pyrophosphate-dependent nucleoside kinase activity was the most variably detected activity. None of the Mycoplasma spp. except four of eight strains of M. hominis had this kinase activity. Likewise, U. urealyticum did not have the pyrophosphate-dependent nucleoside kinase activity; however, A. intermedium and S. citri did have this enzyme activity. The absence of deoxyuridine triphosphatase activity in all Mycoplasma spp. may be related to their proposed rapid evolution and the relative lack of conserved sequences in their 5S ribosomal ribonucleic acids.

Yersinia enterocolitica biogroups 3A and 3B are biochemically, serologically, and ecologically different from biogroup 3 and other Y. enterocolitica biogroups. Both biogroup 3A and biogroup 3B can be characterized by their negative Voges-Proskauer reactions and positive reactions in tests for pyrazinamidase, acid production from mucate, proline peptidase, and acid production from d-xylose. Biogroup 3A ferments l-sorbose but not l-fucose; biogroup 3B has the opposite fermentation pattern. Deoxyribonucleic acid relatedness studies (hydroxyapatite method) indicated that biogroups 3A and 3B are two new species that are about 55% interrelated and 25 to 46% related to other Yersinia species (except Yersinia ruckeri [20 to 22%]). The names Yersinia mollaretii sp. nov. and Yersinia bercovieri sp. nov. are proposed for biogroups 3A and 3B, respectively.

A comparison of 16S ribosomal ribonucleic acid sequences showed that strains classified as Bacteroides succinogenes are not closely related to other species of Bacteroides, including the type species, Bacteroides fragilis. Flavobacterium heparinum, for example, is more closely related to B. fragilis than B. succinogenes is. Therefore, we propose that B. succinogenes strains be renamed as members of a new genus, Fibrobacter. An analysis of 16S ribosomal ribonucleic acid sequence data revealed that there are two species within this new genus. Isolates from rumina are placed in Fibrobacter succinogenes; the type strain is strain S85 (= ATCC 19169). Isolates from ceca of nonruminant animals are placed in Fibrobacter intestinalis; the type strain is strain NR9 (= ATCC 43854). Members of Fibrobacter succinogenes can be differentiated from Fibrobacter intestinalis by their requirement for biotin; the site of isolation may not be diagnostic. Fibrobacter succinogenes consists of two subspecies; the cells of Fibrobacter succinogenes subsp. succinogenes strains are broad rods that are often pleomorphic and coccoid (type strain, S85), whereas cells of Fibrobacter succinogenes subsp. elongata are slender rods (type strain, HM2 [= ATCC 43856]).

A new coagulase-positive species of the genus Staphylococcus, Staphylococcus delphini, is described on the basis of a study of two strains isolated from purulent skin lesions of dolphins. The new species is established and differentiated from the other coagulase-positive Staphylococcus species primarily on the basis of its deoxyribonucleic acid-deoxyribonucleic acid hybridization relationships, its cell wall composition, its bacteriolytic activity pattern, its penicillin-binding protein profile, its biochemical reactions, and the relatively high guanine-plus-cytosine content of its deoxyribonucleic acid. The type strain is strain Heidy (= DSM 20771).

Strains from the intestines of pigs which were originally identified as Streptococcus salivarius were found to be distinct from this species and from other streptococci associated with the intestines of animals. The name Streptococcus hyointestinalis is proposed for the new species. The type strain is strain S93 (= DSM 20770).

The results of deoxyribonucleic acid-deoxyribonucleic acid homology studies and an investigation of physiological properties confirmed the relationship of Streptococcus morbillorum and Gemella haemolysans at the genus level. This is in accord with previously reported data from 16S ribonucleic acid cataloging. The results of this study, together with previously described chemotaxonomic and biochemical properties, indicate that S. morbillorum should be transferred to the genus Gemella as Gemella morbillorum comb. nov.