- Volume 37, Issue 4, 1987

Biochemical and chemical studies were performed on some atypical lactobacilli from chicken meat in an attempt to clarify their taxonomy. The present study showed that the majority of the poultry strains could be allocated to the species Brochothrix thermosphacta, Lactobacillus divergens, and Lactobacillus piscicola. The majority of the remaining poultry strains fell into two distinct groups which were worthy of separate species status. Lactobacillus carnis was found to be a member of the species L. piscicola. On the basis of biochemical, physiological, and chemical criteria, we suggest that L. divergens, L. piscicola, and the two unidentified poultry taxa be classified in a new genus, Carnobacterium, as Carnobacterium divergens comb. nov., Carnobacterium piscicola comb. nov., Carnobacterium gallinarum sp. nov., and Carnobacterium mobile sp. nov. The type strains of C. gallinarum and C. mobile are NCFB 2766T and NCFB 2765T, respectively.

Armadillos are used for propagation of Mycobacterium leprae and are hosts of mycobacteria of the ADM (armadillo-derived mycobacteria) group. The deoxyribonucleic acids (DNAs) of isolates of the five phenetic subgroups of the ADM were analyzed and compared with the genomes of related bacteria. The guanine-pluscytosine (G+C) contents of the DNAs were 62.2 to 67.1 mol% for different ADM strains, 56 mol% for M. leprae and LDC (leprosy-derived corynebacteria), and 62 to 71 mol% for reference mycobacteria. Restriction analysis showed neither adenine methylation in the GATC site (a specific trait of LDC strains) nor cytosine methylation in the CCGG and GGCC sites. Hybridization higher than 80% was obtained with DNA from isolates within the same ADM subgroups, whereas 17 to 50% hybridizations were observed with organisms from different ADM subgroups. Genomes of ADM strains and reference mycobacteria were 15 to 40% homologous, except for subgroups IV and V, whose DNAs were 54 to 62% homologous with Mycobacterium lepraemurium DNA. Little or no homology between M. leprae and ADM genomes was found. We concluded that single ADM subgroups can be considered as distinct species within the genus Mycobacterium; they are genetically unrelated to the other leprosy-associated bacteria.

A nonobligate, bacterial predator of bacteria in soil is described. It was previously designated as strain N-1. It is a gram-negative, aerobic, mesophilic, short rod that multiplies by binary fission. It is motile and has peritrichous flagella. It has simple nutritive requirements when not acting as a predator. Glucose is not utilized as a carbon source. Instead, it uses fructose or any of several amino acids. As a predator, it attacks both gram-positive and gram-negative bacteria, including certain other nonobligate bacterial predators of bacteria in soil. It is highly resistant to copper, and its growth initiation is strongly stimulated by copper. Copper has no effect on the cellular multiplication that occurs after growth has initiated. A new genus, Cupriavidus, is proposed for this bacterium. The type species is Cupriavidus necator sp. nov., and strain N-1 (ATCC 43291) is the type strain of this species.

Yersinia rohdei sp. nov vaisolated from feces of dogs and humans and from surface water. The organisms gave the key biochemical reactions typical of Yersinia species, including greater metabolic activity at 25 to 28°C than at 36°C. Y. rohdei can be distinguished from other yersiniae by its positive reactions in tests for citrate and sucrose and negative reactions in tests for indole, acetoin (Voges-Proskauer), and rhamnose. Two biogroups were found among seven strains investigated, one of which fermented raffinose and melibiose. Deoxyribonucleic acid hybridization tests (hydroxyapatite method) showed average levels of relatedness of 93% at a 60°C incubation temperature and 97% at 75°C. Labeled Y. rohdei deoxyribonucleic acid showed 30% to 63% relatedness to all Yersinia species except Yersinia ruckeri. The guanine-plus-cytosine contents of five tested strains were 48.7 to 49.4 mol%. All strains possessed O antigens not typable with antisera of the extended typing scheme of Wauters, but most of them had H antigens typical of Yersinia enterocolitica biotype 1. Five tested strains belonged to lysotype Xz. Y. rohdei strains were susceptible to most antimicrobial agents; resistance or intermediate susceptibility was observed for ampicillin, carbenicillin, cephalothin, penicillin, and sulfonamide. Calcium dependency, autoagglutination, and Congo red pigmentation tests, which are indicative of the presence of a virulence plasmid to Y. enterocolitica, were negative in Y. rohdei. The clinical significance for humans and animals is uncertain and requires further study. The type strain of Y. rohdei sp. nov. is strain H 271-36/78 (= CDC 3022-85 = ATCC 43380).

Seven ureaplasma strains isolated from the oropharynx of chickens (Gallus gallus var. domesticus) were characterized and compared with the two previously described species of this genus, Ureaplasma urealyticum from humans and Ureaplasma diversum from cattle. The avian isolates were found to have similar biological, biochemical, serological, and genomic properties and were readily distinguished from the two previously described species. These avian ureaplasma strains have been reported to hydrolyze urea, but not to catabolize arginine or glucose; they lack a cell wall, pass through a 450-nm membrane filter, require cholesterol for growth, and form minute colonies (15 to 60 μm) on agar medium. The seven avian strains constitute a homogeneous group based on their antigenic properties as determined by the metabolism and growth inhibition tests and by protein composition analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. On the basis of these findings, we propose that ureaplasmas with these characteristics belong to a new species, Ureaplasma gallorale, with strain D6-1 (= ATCC 43346 = NCTC 11707) as the type strain.

The species Lactobacillus pentosus was omitted from the Approved Lists of Bacterial Names. Nucleic acid and biochemical studies demonstrated that Strains presently designated L. pentosus represent a distinct taxon worthy of separate specific status. It is therefore formally proposed that the species L. pentosus be revived. The type strain of L. pentosus is NCDO 363.

A new species of Actinomadura, for which we propose the name Actinomadura atramentaria, was isolated from a soil sample collected in Japan. Whole-cell hydrolysates contain meso-diaminopimelic acid and a trace of madurose. MK-9(H6) is the major menaquinone, and tuberculostearic acid is the major fatty acid. No nitrogenous phospholipids or mycolic acids are present. A. atramentaria is characterized by its white aerial masses of thickly tufted short chains of two to five spores (mostly two to three spores) with smooth surfaces and by the formation of inky brown diffusible pigments. The type strain of A. atramentaria is strain SF2197 (= JCM 6250).

Several strains of halotolerant bacteria from North America were compared for 125 characteristics. Although these organisms had several characteristics in common, they are a relatively heterogeneous group. The 12 strains could be divided into at least three and perhaps as many as five taxa. Halomonas elongata ATCC 33173T (T = type strain) and “Pseudomonas halodurans” ATCC 29686T had 79% phenotypic similarity, indicating that these organisms are probably separate species of the same genus. We conclude that this relationship is sufficient to warrant assignment of “Pseudomonas halodurans” to the genus Halomonas as Halomons halodurans sp. nov., nom rev., comb nov.

Crossed immunoelectrophoresis (XIE) reference systems were established for Fluoribacter (Legionella) (containing Fluoribacter bozemanae, Fluoribacter dumoffii, and Fluoribacter gormanii) and for Tatlockia (Legionella) micdadei. The Fluoribacter and Tatlockia XIE reference systems contained 54 and 72 anodemigrating antigens, respectively. These two systems, together with the previously described polyvalent Legionella pneumophila (serogroups 1 through 6) XIE reference system, were used to study the crossreactivities of antigens from organisms comprising the three proposed genera in the family Legionellaceae. Antigenic homology was expressed as the matching coefficient (MC), the ratio of the number of cross-reactive antigens to the total number of antigens. The MCs for individual L. pneumophila serogroups when the polyvalent L. pneumophila antibody was used were 0.98 ± 0.05, which was significantly higher than the MCs determined by using Fluoribacter or Tatlockia antibodies (0.50 ± 0.13) (P ≪ 0.001). The MCs for the three species of Fluoribacter when polyvalent Fluoribacter antibody was used were 0.93 ± 0.10, which was also significantly higher than the MCs when heterologous antibodies were used (0.40 ± 0.04) (P ≪ 0.001). The MCs for T. micdadei with the two heterologous antibody preparations were similar to each other (0.32 and 0.46) and to all other heterologous MCs among members of the Legionellaceae. The MCs for organisms representing three other families of bacteria were 0.16 ± 0.04 in all three XIE reference systems and were significantly lower than the MCs among members of the Legionellaceae (P ≪ 0.001). When a priori criteria for MC interpretation established in previous serotaxonomic studies of other bacterial species by XIE were used, our results from studies on the antigenic relationships among Legionella, Fluoribacter, and Tatlockia supported the proposal that there are multiple genera in the family Legionellaceae.

Fourteen spiroplasma strains, primarily of insect origin, were analyzed according to criteria previously proposed for description of new serogroups of the genus Spiroplasma. When tested by reciprocal metabolism inhibition, growth inhibition, and deformation serological procedures, 12 of the strains were serologically unrelated to each other and to representative strains previously assigned to groups I to XI and subgroups I-1 to I-8. Examination by dark-field and electron microscopy indicated that each of the 12 strains possessed morphological features typical of spiroplasmas (helicity, motility, lack of a cell wall, and absence of periplasmic fibrils). All strains were resistant to 500 U of penicillin per ml and catabolized glucose but were unable to hydrolyze urea. Ability to hydrolyze arginine varied among strains. The guanine-plus-cytosine contents of the deoxyribonucleic acid of the 12 strains varied from 24 to 29 mol%. Two other strains (MQ-6 and Ar-1357) shared only a partial serological relationship to strain CC-1 (group XVI), suggesting that this group may consist of an assemblage of heterogeneous serovars. On the basis of the unique serological distinctions and other properties reported herein, we propose that the 12 representative strains be assigned new consecutive group designations XII to XXIII.

Spiroplasma strain AES-1, which was isolated from the salt marsh mosquito Aedes sollicitans, is the monotypic representative of group X. This strain differed in serological reactivity, growth pattern, and morphology from previously described spiroplasmas. Most cells of strain AES-1 were nonhelical; helical cells were about 1 to 2 μm long and rarely had more than one or two turns. Optimal growth occurred at 31°C; no growth was observed at 37°C. Cholesterol and amino acids were required for growth. Strain AES-1 utilized glucose, fructose, mannose, mannitol, and trehalose in the chemically defined CC-494 medium, failed to hydrolyze arginine and urea but reduced tellurite either aerobically or anaerobically, and was susceptible to digitonin. Polyacrylamide gel electrophoretic patterns of the cell proteins of strain AES-1 were distinctive and were characterized by a major low-molecular-weight protein band. The guanine-plus-cytosine content was 26.6 mol% as determined by thermal denaturation. Serological studies, using growth inhibition, deformation, and metabolism inhibition tests, showed that strain AES-1 was distinct from all recognized spiroplasma groups and subgroups, including the six named Spiroplasma species; avidin-biotin-peroxidase enzyme-linked immunosorbent assays showed that strain AES-1 was unrelated to S. citri, S. melliferum, S. kunkelii, S. floricola, and S. apis. Strain AES-1 is the first to be characterized biochemically in a chemically defined medium. We propose that strain AES-1 (ATCC 35112) be designated as the monotypic representative of a new species, Spiroplasma culicicola sp. nov.

Estimates of deoxyribonucleic acid (DNA) relatedness and phenotypic characterization of 98 Bacillus polymyxa strains revealed that 70 (groups 1A and 1B) were closely related to the B. polymyxa type strain and 28 (group 2) were not. Group 1A strains (29) showed 80 to 100% DNA relatedness to the type strain, while the remaining 41 strains (group 1B) gave reduced relatedness values of 57 to 68%. Groups 1A and 1B were phenotypically identical. Low DNA homology values indicated that group 2 strains were not closely related to group 1A or 1B strains or to the type strains of Bacillus species that have guanine-plus-cytosine contents of 44 to 53 mol% or form gas from carbohydrates or both, namely, B. alvei, B. badius, B. brevis, B. coagulans, B. licheniformis, and B. macerans. Group 2 strains were phenotypically distinct from B. polymyxa and the seven recognized species.

A new saccharolytic species of Clostridium was isolated from a slurry of rat cecal contents and is described. The single strain isolated often forms cells with two spores. On the basis of several major biochemical characteristics it differs from the only other clostridium species known to form cells with two spores, C. oceanicum. The name Clostridium disporicum is proposed. Type strain DS1 is deposited in the National Collection of Industrial Bacteria as strain NCIB 12424.

The marine methanol-utilizing bacteria in the genus Methylophaga are obligately methylotrophic, but some strains utilize d-fructose in addition to methanol. The deoxyribonucleic acid base composition of these organisms ranges from 43.0 to 49.0 mol% guanine plus cytosine. Their cellular fatty acids consist predominantly of large amounts of straight-chain saturated C16:0 acid and unsaturated C16:1 acid. Methylophaga strains are distinguished from Methylobacillus strains by their tolerance to sodium chloride and their deoxyribonucleic acid base composition and from Methylomonas and Methylococcus strains by their utilization of methane, their tolerance to sodium chloride, and their deoxyribonucleic acid base composition.

Deoxyribonucleic acid hybridization (hydroxyapatite method, 55 and 70°C) was used to characterize 38 serovars from 22 named serogroups of Leptospira interrogans and Leptospira biflexa, 6 serovars from 4 new unnamed serogroups of Leptospira interrogans, and single serovars of the proposed species Leptospira parva and Leptonema illini. Deoxyribonucleic acid relatedness confirmed the validity of Leptospira parva and Leptonema illini. The well-accepted species Leptospira interrogans and Leptospira biflexa, as currently defined, were extremely heterogeneous. Relatedness results revealed at least five new species among the parasitic serovars formerly included in Leptospira interrogans and two new species among the saprophytic serovars formerly included in Leptospira biflexa. Serogrouping did not equate with species identification, as serovars from several different subserogroups belonged to different species. The new species named in this paper are Leptospira, noguchii, Leptospira weilii, Leptospira santarosai, Leptospira borgpetersenii, Leptospira meyeri, Leptospira wolbachii, and Leptospira inadai.

Eleven strains of gram-negative, denitrifying, prodigionine-producing marine bacteria with sheathed flagella were recovered from the fjord system off the Norwegian west coast. These isolates form a physiologically and morphologically very homogeneous group. Apart from their ability to denitrify and having sheathed flagella, the strains correspond closely to the current description of the genus Alteromonas. Their strictly respiratory metabolism precludes their inclusion in the genus Vibrio, and the low guanine-plus-cytosine contents of their deoxyribonucleic acids preclude their inclusion in the genus Pseudomonas or the genus Marinomonas. Despite their ability to denitrify, we suggest that these organisms belong to the genus Alteromonas and constitute a new species, Alteromonas denitrificans. A detailed description of the type strain of Alteromonas denitrificans, Nygaard 1977, is given. The type strain has been deposited with the American Type Culture Collection, Rockville, Md., as strain ATCC 43337.

Thirty-four strains of Xanthomonas ampelina, the causal agent of bacterial necrosis of grape vines, were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of their cellular proteins and by numerical analysis of 106 enzymatic features (API systems). These organisms formed a very homogeneous taxon. Generic and suprageneric relationships were determined by hybridizations between 23S 14C-labeled ribosomal ribonucleic acid from Xanthomonas ampelina NCPPB 2217T (T = type strain) and deoxyribonucleic acids from eight Xanthomonas ampelina strains, Xanthomonas campestris NCPPB 528T, and the type strains of 16 possibly related species. Xanthomonas ampelina was found to be a totally separate subbranch in ribosomal ribonucleic acid superfamily III, without any relatives at the generic level. It is not related to the genus Xanthomonas. Genetically its closest relatives are, among others, Pseudomonas acidovorans, Alcaligenes paradoxus, and Comamonas terrigena. We propose the transfer of Xanthomonas ampelina to a new genus, Xylophilus. The only species and thus the type species is Xylophilus ampelinus. The type strain is NCPPB 2217.

In a preliminary investigation the isolation of a caldoactive filamentous microorganism from a New Zealand hot spring was reported. This organism is described here as a new species belonging to the genus Thermus, namely, Thermus filiformis, based on ultrastructural, phenotypic, and anomalous Gram type characteristics. The cell wall of T. filiformis resembles that of Thermus aquaticus apart from the presence of an extra layer. The Thermus species tested, including T. filiformis, are negative for the aminopeptidase test, which is unusual for a gram-negative genus. T. filiformis is nonproteolytic, unlike most other Thermus strains, and also differs radically from other strains in morphology when it is observed by using phase-contrast microscopy. The single strain of the species has been deposited with the American Type Culture Collection as strain ATCC 43280T (T = type strain).

Relatedness among strains of Lactobacillus buchneri, Lactobacillus brevis, Lactobacillus hilgardii, Lactobacillus confusus, Lactobacillus fermentum, Lactobacillus cellobiosus, Lactobacillus reuteri, Lactobacillus collinoides, Lactobacillus fructivorans, Lactobacillus kandleri, and Lactobacillus viridescens was determined by computerized comparison of total soluble cell protein patterns. Two related subgroups of L. buchneri were obtained: the first comprised a tight cluster of strains ATCC 11579 and ATCC 9460 with neotype strain ATCC 4005, and the second contained strains ATCC 12936, ATCC 12935, and ATCC 11307. L. brevis ATCC 13648, ATCC 4006, ATCC 367, and ATCC 8287 appear to be related to the neotype strain of L. brevis (ATCC 14869), L. brevis ATCC 11577 to L. hilgardii, and L. confusus ATCC 27646 to L. brevis ATCC 367 and ATCC 8287. L. brevis ATCC 14434 and L. cellobiosus ATCC 11740 were originally misclassified. The protein patterns of these strains did not resemble those of any of the other named lactobacilli which were examined. Our findings support the reclassification of L. brevis ATCC 8007 as Lactobacillus kefir, L. cellobiosus ATCC 11739 as L. fermentum, and L. fermentum ATCC 23272 as L. reuteri. L. collinoides, L. fructivorans, L. hilgardii, L. kandleri, L. reuteri, and L. viridescens are distinct, separate species.