Capnocytophaga: New genus of Gram-negative gliding bacteria. III. Physiological characterization

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268

186

We compared 46 strains of gram-negative, asaccharolytic, rod-shaped bacteria which were isolated from humans with gingivitis, periodontal pockets, and lesions in alveolar bone with 10 reference strains of Eikenella corrodens, Vibrio succinogenes, Bacteroides ureolyticus, and species of Campylo bacter. We divided these 56 strains into seven groups based on the guanine-plus-cytosine contents of their deoxyribonucleic acids, their deoxyribonucleic acid homologies, and cluster analyses of their phenotypic features. A total of 23 of the fresh isolates showed more than 90% similarity (Jaccard coefficient) with E. corrodens. Growth of the remaining 23 isolates was enhanced in broth cultures by formate and fumarate.These isolates were not members of B. ureolyticus, V. succinogenes, or previously described species of Campylo bacter; they constituted three distinct new species. We propose Bacteroides gracilis sp. nov. (type strain, ATCC 33236) as the name for seven isolates of slender, gram-negative, nonmotile, anaerobic, rod-shaped bacteria that corroded agar and had deoxyribonucleic acid guanine-plus-cytosine contents of 44 to 46 mol%. All of the remaining isolates were motile by means of a single polar flagellum. Ten anaerobic strains were similar to V. succinogenes in phenotypic characteristics and guanine-plus-cytosine contents. However, these strains were distinct from V. succinogenes on the basis of deoxyribonucleic acid homology results. We propose Wolinella as the name of a new genus to include anaerobic, asaccharolytic, rod-shaped bacteria with single polar flagella and deoxyribonucleic acid guanine-plus-cytosine contents of 42 to 49 mol%. Wolinella succinogenes (Wolin et al.) comb. nov. is designated the type species of the genus, and ATCC 29543 is the type strain of W. succinogenes. We propose Wolinella recta sp. nov. (type strain, ATCC 33238) as the name for nine of the strains that formed a related but distinct group. We propose Campylobacter concisus sp. .nov. (type strain, ATCC 33237) as the name for the six isolates of noncorroding, microaerophilic, gram-negative, rod-shaped bacteria that have predominantly curved cells and deoxyribonucleic acid guanine-plus-cytosine contents of 34 to 38 mol%. The description of the genus Campylobacter is amended to include species with deoxyribonucleic acid guanine-plus-cytosine contents of 30 to 38 mol%.We isolated gram-negative, asaccharolytic, rod-shaped bacteria from humans with lesions of advanced destructive periodontal disease; the majority of these bacteria pitted or corroded agar media. These organisms were nonmotile, or they exhibited either cell "twitching" or active darting motility when they were examined by dark-field microscopy. These isolates were numerically dominant in some active disease sites (31). They were difficult to separate into distinct groups on the basis of routine characterization studies, and some of them were different from isolates of known species. In this investigation we compared strains isolated from humans with advanced periodontal d...

Section: Materlals and Methodsmentioning

confidence: 99%

Section: Materlals and Methodsmentioning

confidence: 99%

Wolinella gen. nov., Wolinella succinogenes (Vibrio succinogenes Wolin et al.) comb. nov., and Description of Bacteroides gracilis sp. nov., Wolinella recta sp. nov., Campylobacter concisus sp. nov., and Eikenella corrodens from Humans with Periodontal Disease

Tanner

Badger

Lai

et al. 1981

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268

186

“…Cells of reference strains were grown in 2-to 3-liter broth cultures (mycoplasma broth [BBL] supplemented with 5 mg of hemin per liter) for extraction of DNA. The DNA was extracted by using a method modified (13) from the procedure of Marmur (9). G+C contents were determined by the thermal denaturation method (1).…”

mentioning

confidence: 99%

Bacteroides forsythus sp. nov., a Slow-Growing, Fusiform Bacteroides sp. from the Human Oral Cavity

Tanner¹,

Listgarten²,

Ebersole³

et al. 1986

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175

150

The characteristics of a group of slow-growing, fusiform, fastidious anaerobes isolated from advanced periodontal lesions in human oral cavities were examined. Our results indicated that 12 fusiform Bacteroides strains belong to a new species in the genus Bacteroides. The name Bacteroides forsythus is proposed for these isolates. The type strain is strain ATCC 43037.Anaerobic, gram-negative, fusiform organisms which did not resemble previously recognized speqies were isolated from deep periodontal pockets of humans that showed progressive tissue loss (3,15,17). The cells appeared to be long filaments or medium rods with tapered (fusiform) or rounded ends. Some cells had central swellings. Large spheroids were frequently associated with the cells, particularly the cells with filamentous morphology. These strictly anaerobic fusiform Bacteroides strains grew slowly on blood agar plates and grew poorly in all broth media tested and on most agar media tested. Biochemical tests gave inconsistent results and were usually negative. For this reason, the fusiform Bacteroides strains were characterized without using most of the conventional biochemical tests, Deoxyribonucleic acid (DNA) guanine-plus-cytosine (G +C) content, DNA-DNA homology, cell wall ultrastructure, serology, API enzyme substrate tests (Analytab Products, Plainview, N.Y.), and protein profiles of whole sonicated cells obtained by using polyacrylamide gel electrophoresis (PAGE) indicated that these organisms comprise a distinct species in the genus Bacteroides. The name Bacteroides forsythus is proposed for this new species.Strains and inocula. The strains which we used are listed in Table 1. These organisms were maintained on Trypticase soy agar plates supplemented with 5% sheep blood (TSBA; BBL Microbiology Systems, Cockeysville, Md.) at 35 to 37°C in an atmosphere containing 10% H2, 10% CO2, and 80% N2. The fusiform Bacteroides strains were maintained on TSBA plates; a fusiform Bacteroides strain was streaked on one-half of each plate, and a strain of Fusobacterium nucleaturn was streaked on the other half of the plate. The strains were stored in liquid nitrogen by using previously described methods (14). Inocula for tests were obtained by scraping cells from the surfaces of TSBA plates. At least 100 plates of fusiform Bacteroides strains were used to obtain sufficient cells for DNA extraction. Two to four plates were used to provide the inoculum for an enzyme-linked immunosorbent assay and for each of the tests in the API enzyme substrate test series.DNA-DNA hybridization. Cells of reference strains were grown in 2-to 3-liter broth cultures (mycoplasma broth [BBL] supplemented with 5 mg of hemin per liter) for extraction of DNA. The DNA was extracted by using a method modified (13) from the procedure of Marmur (9). G+C contents were determined by the thermal denaturation method (1). Levels of DNA-DNA homology were determined by using the initial renaturation rates (2).API enzyme substrate tests. API ZYM and API An-Ident enzymatic substrate tests were ...

“…Two-to t hree-liter broth cultures were grown to log phase in PPLO-FF medium and harvested by centrifugation. DNA was isolated by a modification (19) of the Marmur method (11). DNA solutions were dissolved in a phosphate buffer containing 9.47 g of Na2HP04 per liter and 9.2 g of NaH2P04 per liter.…”

mentioning

confidence: 99%

Wolinella curva sp. nov.: "Vibrio succinogenes" of Human Origin

Listgarten

Ebersole

1984

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Asaccharolytic, anaerobic vibrios which require formate and fumarate for growth in broth culture have been isolated from humans. Some of these strains resemble Wolinella succinogenes phenotypically, but show no deoxyribonucleic acid (DNA)-DNA homology with either W . succinogenes or Wolinella recta. In this investigation, six unidentified strains were compared with reference and type strains of W . recta and W . succinogenes by using cluster analysis of phenotypic characteristics, enzyme-linked immunosorbent assays, and cell wall ultrastructure. One unidentified strain, a sewage isolate, was W . succinogenes. The remaining unidentified strains showed no DNA-DNA homology with either W . succinogenes or W . recta. Wolinella curva sp. nov. is proposed for four isolates which were distinct, as determined by serological and DNA-DNA homology experiments, from the previously described species. W . curva strains could also be differentiated from W. recta by morphology and by the growth of W. curva in the presence of Janus green (0.1 g/liter), basic fuchsin (0.032 ghiter), sodium deoxycholate (1.0 g/liter), indulin scarlet (0.5 g/liter), oxgall (10 g/liter), safranine (0.5 g/liter), azure I1 (0.025 g/liter), penicillin (16 pg/ml), and polymixin B (4 pg/ml). W . curva and W . succinogenes are similar in cell ultrastructure and in other phenotypic features.Wolinella succinogenes (Vibrio succinogenes) was isolated from a bovine rumen by Wolin et al. (24). Members of this species use formate or hydrogen as an electron donor. Fumarate was found to be the most efficient electron acceptor of several organic and inorganic compounds tested (1, 24). Two other motile, metabolically similar species from humans have been described; these are Wolinella recta, with a guanine-plus-cytosine (G+C) content of 44 to 46 mol%, and Campylobacter concisus, with a C+G content of 34 to 38 mol% (20).Several investigators have isolated from human sources anaerobic, gram-negative, asaccharolytic, motile vibrioshaped rods with metabolism similar to that of W . succinogenes (17,20,21,23). Deoxyribonucleic acid (DNA)-DNA homology techniques have shown that these additional strains are distinct from previously described species (21). In this investigation we determined the taxonomic status of additional formate-and fumarate-requiring strains isolated from humans. MATERIALS AND METHODSCharacterization. The origins of the oral and reference strains which we used are shown in Table 1. The methods used to maintain and characterize these strains were similar to those described previously (20). Cell morphology and motility were determined by dark-field microscopy of young broth cultures. Biochemical tests were performed in broth cultures and on agar plates. The basal medium used for broth tests (PPLO-FF medium) contained (per liter) 21 g of Mycoplasma broth base (BBL Microbiology Systems, Cockeysville, Md.), 5 mg of hemin, 2 g of sodium formate, and 3 g of sodium fumarate and was adjusted to pH 7.4. A 30-ml portion of a log phase inoculum was added to 100 ml ...