A group of Eubacterium-like strains (designated group S14), isolated from the feces of healthy people, was characterized by biochemical tests, fatty acid analysis, cell wall murein analysis, and 16S rDNA analysis. Our results indicate that group S14 is phylogenetically a member of the Clostridium subphylum of the gram-positive bacteria. Despite a phenotypic resemblance to the genus Eubacterium, group S14 was shown to be phylogenetically distantly related to the type species of the genus, Eubacterium limosum. Group S14 showed a specific phylogenetic association with Erysipelothrix rhusiopathiae. Group S14 resembled Erysipelothrix in possessing the uncommon type B cell wall murein. Structural analyses, however, revealed the presence of a previously unknown B1 delta (L-Ala)-D-Glu-Gly-L-Lys murein type. Based on a 16S rRNA sequence divergence of greater than 10% with E. rhusiopathiae and the presence of a unique murein type, a new genus, Holdemania, is proposed for group S14, with one species, Holdemania filiformis. Type strain of H. filiformis is ATCC 51649.

The fecal floras of polyp patients, Japanese-Hawaiians, North American Caucasians, rural native Japanese, and rural native Africans were compared. The polyp patients and Japanese-Hawaiians were considered to be groups at high risk of colon cancer, and the rural native Japanese and rural native Africans were considered to be groups at low risk. The North American Caucasians were found to have a flora composition intermediate between these two groups. Fifteen bacterial taxa from the human fecal flora were significantly associated with high risk of colon cancer, and five were significantly associated with low risk of colon cancer. Total concentrations of Bacteroides species and, surprisingly, Bifidobacterium species were generally positively associated with increased risk of colon cancer. Some Lactobacillus species and Eubacterium aerofaciens, which also produces major amounts of lactic acid, showed closest associations with low risk of colon cancer.

Clostridium putrificum (Trevisan 1889) Reddish and Rettger 1922; Clostridium botulinum (van Ermengem 1896) Bergey, Harrison, Breed, Hammer, and Huntoon 1923; and Clostridium sporogenes (Mechnikoff 1908) Bergey, Harrison, Breed, Hammer, and Huntoon 1923 are genetically related at the species level. We propose rejection of the name C. putrificum (which has priority) and conservation of the name C. botulinum on the basis of Rules 23a and 56a of the International Code of Nomenclature of Bacteria and conservation of the name C. sporogenes for nontoxigenic strains according to Rules 23a and 56b.

Cellular fatty acid profiles were determined for species in 33 genera of anaerobic gram negative bacilli and were confirmed to be a useful taxonomic tool. Most of the genera could be differentiated by visual inspection of their profiles. The three genus pairs that were most difficult to distinguish visually (Bacteroides and Prevotella, Pectinatus and Megamonas, and Serpulina and Bilophila) and the species of these genera were differentiated by the MIDI (Microbial ID, Inc.) identification system. Similarities in cellular fatty acid profiles may be correlated with similarities in other phenotypic characteristics, but more often there is no other obvious phenotypic relationship. Although medium components may not change the constituents detected or the ratios among the constituents detected for some species, identical medium changes may result in vast differences in the profiles obtained with other species. Thus, if a worker wishes to compare profiles of various taxa, it is essential that the same cultural and analytical conditions be used.

Prevotella tannerae sp. nov. and Prevotella enoeca sp. nov. from the human gingival crevice are described. These organisms are obligately anaerobic, non-spore-forming, nonmotile, gram-negative, rod-shaped bacteria that ferment carbohydrates and produce succinic and acetic acids. Bile inhibits growth. Some strains (38%) of P. tannerae produce colonies with a tanto black pigment when they are grown on rabbit blood agar. The type strains are P. tannerae ATCC 51259 and P. enoeca ATCC 51261. In addition, the description of Prevotella zoogleoformans is emended to exclude strains now recognized as members of Prevotella heparinolytica.

The following four new species of anaerobic gram-negative bacilli isolated from the human gingival crevice are described: Oribaculum catoniae, with ATCC 51270 as the type strain; Catonella morbi, with ATCC 51271 as the type strain; Hallella seregens, with ATCC 51272 as the type strain; and Johnsonella ignava, with ATCC 51276 as the type strain. C. morbi is associated with periodontitis. H. seregens and J. ignava are associated with gingivitis and periodontitis. O. catoniae has been isolated from healthy and diseased gingiva. Dialister pneumosintes (Olitsky and Gates 1921) gen. nov., comb. nov., nom. rev., associated with gingivitis, is proposed to accommodate organisms formerly classified as Bacteroides pneumosintes.

The subgingival microflora of 39 HIV+ subjects with gingivitis or adult periodontitis was cultured quantitatively anaerobically for bacteria, spirochetes, and mycoplasma and aerobically for yeasts. Isolates were characterized by conventional biochemical tests, polyacrylamide gel electrophoresis of soluble proteins, cellular fatty acid profiles, immunofluorescence, and immunodiffusion. In general, the same types of bacteria were isolated from the subgingival crevice of HIV+ subjects as we previously had isolated from the subgingival crevice of non-HIV subjects. A statistically significant difference was found between the composition of the flora of HIV+ subjects with adult periodontitis (AP) and concurrent studies of a non-HIV+ AP population. Mycoplasma salivarium was significantly elevated in the HIV+ subjects examined. Yeasts were isolated from only 10% of the samples and from 13% of the HIV-positive subjects at 0.05 to 0.0002% of the total cultivable count when present.

The classical twin model was utilized in this study in an attempt to determine the importance of host genetics to the composition of the subgingival flora. Simultaneously, the effect of puberty on the flora composition was assessed. The compositions of the floras were significantly different at ages 11 and 14 in the same people, indicating that transition to an adult flora composition may be initiated during puberty. However, the numbers of subjects who had prepubertal and postpubertal testosterone levels in this study were too small to demonstrate significant differences based solely on testosterone level (P = 0.053 and 0.11 for tests of unrelated members, i.e., all twins "a," the first twin of each pair, and all twins "b," the second twin of each pair). Sixteen unrelated 11-year-old subjects had prepubertal levels of less than 30 ng of testosterone per dl of serum, and only six of these unrelated subjects had levels above 300 ng/dl by age 14. Of their twin siblings, who formed the second group of unrelated individuals, 15 had prepubertal levels and only 5 reached postpubertal levels. Unpaired t tests indicated that Veillonella atypica, Prevotella denticola, and Prevotella melaninogenica were among the species that contributed most to changes in flora composition during puberty. The compositions of subgingival floras of 11-year-old monozygous and dizygous male twins were significantly more similar than those of unrelated subjects in the study (P = 0.004 and 0.009, respectively). At 12.5 years of age, the floras of monozygous twins remained more similar than those of unrelated subjects (P = 0.001), but the dizygous-twin floras were not significantly more similar than those of unrelated people. This difference corresponded with moderate and varied testosterone levels within dizygous-twin pairs at age 12.5. By age 14 both monozygous and dizygous twins again had floras with compositions more similar than those of unrelated people (P = 0.008 and 0.002, respectively). Estimates of the genetic contributions to the increased similarity of the floras of twins as compared with floras of unrelated people indicated that the concentrations of several species in the flora may be influenced by host genetic factors. The prevalence of certain other species appeared to be controlled primarily by environment.

Previous studies have demonstrated that demographic characteristics of subject populations influence both the incidence of periodontal diseases and various aspects of host responses to periodontal bacteria. In this study we analyzed the components of the subgingival microflora from individuals with adult periodontitis, early onset periodontitis, gingivitis, and periodontal health as a function of gender and race (black and white). Clinical categories were analyzed individually so that there were no differences in the clinical characteristics of the sampled sites. No significant differences were noted in the subgingival microflora between males and females. When either the first two bacterial samples from each subject or all bacterial samples taken from each subject were included in the analysis, it was found that Porphyromonas gingivalis was more significantly associated with black subjects in the adult periodontitis group. When all samples were considered in the analysis, it was found that Peptostreptococcus anaerobius was associated with black subjects in the adult periodontitis group, while Fusobacterium nucleatum was associated with white subjects in both the adult periodontitis and early onset periodontitis groups. Thus a limited number of important bacterial components of the subgingival microflora are influenced by the race and diagnosis of the subject group.

During studies of human periodontal disease, a number of bacterial strains were encountered that, on the basis of results of standard biochemical tests, appeared to be Prevotella buccalis, Prevotella denticola, Prevotella melaninogenica, or Prevotella loescheii. However, use of the standard biochemical tests, cellular fatty acid analyses, and the polyacrylamide gel electrophoresis patterns of soluble proteins resulted in conflicting identifications of these strains. The results of tests for cellobiose fermentation, inulin fermentation, and pigment production were responsible for most of the discordant results. Cellular fatty acid analyses in which the Microbial Identification System was used did not differentiate these strains from validly described species, even though separate library entries were created for them. DNA reassociation determinations in which the S1 nuclease procedure was used showed that cellobiose fermentation and pigment production are variable among strains of P. melaninogenica and P. denticola and that fermentation of xylan is not a reliable characteristic for differentiating P. buccalis from Prevotella veroralis. In contrast to previous indications, most strains of P. veroralis do not ferment xylan. These species can be differentiated by DNA-DNA reassociation and by cellular fatty acid analysis, using the Microbial Identification System, but differentiation by currently described phenotypic characteristics is not reliable. Similarly, P. loescheii and the genetically distinct (but closely related) D1C-20 group cannot be distinguished reliably from each other or from P. veroralis, P. denticola, and P. melaninogenica on the basis of currently described phenotypic tests other than cellular fatty acid composition or, for some species, electrophoretic patterns of soluble whole-cell proteins.

Nonisotopic, whole-genomic DNA probes, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), biochemical tests in microtiter trays and cellular fatty acid (CFA) analysis were compared for the identification of 5 oral Selenomonas species. DNA probes were prepared by biotin-labeling DNA extracted from the type strains of Selenomonas noxia, Selenomonas flueggei, Selenomonas artemidis, Selenomonas infelix and Selenomonas sputigena. The probes were hybridized with DNA from 21 reference strains, 18 fresh isolates of Selenomonas species, and 21 strains of other oral gram-negative species. Target DNAs were obtained by in situ extraction of colonies blotted onto filter paper. Streptavidin-linked alkaline phosphatase was used to detect homologous reactions of probe and target DNA. Each Selenomonas species DNA probe reacted with reference strains of only that species. All Selenomonas strains that reacted with the DNA probe for a particular species gave similar biochemical test results, SDS-PAGE protein profiles, and CFA profiles to those of the type strain of the corresponding species. All the methods tested were useful for identifying the species, and all yielded similar identifications of the fresh isolates. The DNA probes, however, had the potential for identifying Selenomonas species directly from primary isolation plates or plaque samples.

Onthebasis of686analyses of285strains ofClostridium botulinum, Clostridium argentinense (formerly C. botulinum typeG),andphenotypically related organisms, 14cellular fatty acid(CFA)groupsoftoxic organisms and6CFAgroups ofnontoxic organisms weredelineated. TheCFAgroups oftoxic strains included twooftypeA,threeofproteolytic strains oftypeB,twoofproteolytic strains oftypeF,oneeachof nonproteolytic strains oftypes B,E,andF,andoneeachoftypes Ca,CoI, andD andC.argentinense. The groups ofphenotypically similar nontoxic strains included Clostridium sporogenes, Clostridium putrificum, nontoxic strains withphenotypic characteristics similar tothose ofnonproteolytic strains ofC.botulinum types B,E,andF(BEF-like), twogroups ofnontoxigenic organisms withphenotypic characteristics similar tothose ofC.botulinum types C andD andClostridium novyi (CDN-like), andClostridium subterminale, whichhas phenotypic characteristics similar tothose ofC.argentinense. Within thetoxin types, 89to100%ofthestrains werecorrectly identified byCFAanalysis, and74to100%oftheanalyses werecorrect. Of36strains ofC. sporogenes, 30(83%)werecorrectly identified; 17%ofthestrains ofC.sporogenes wereincorrectly identified asC.botulinum typeAorB.Allanalyses ofC.putrificum andC.subterminale werecorrectly identified. There wasnosignificant level ofsimilarity between strains ofC.botulinum andphenotypically similar organisms and 85other species ofclostridia or407other taxaofgram-positive andgram-negative bacteria. Additionally, the onestrain eachofClostridium baratii andClostridium butyricum previously reported toproduce C.botulinum toxin could bedifferentiated fromC.botulinum types aswell asfromstrains ofC.baratii andC.butyricum that didnotproduce aneurotoxin.

20 adult periodontitis (AP) subjects were examined every 2 to 4 months and microbiological samples were collected and cultured when 2 mm or more loss of attachment (active sites) was detected by 2 examiners. Similar sites in which no progressive destruction was observed (control sites) also were sampled in the same subjects. By lambda-analysis, there was no statistically significant difference in floras of active (42 sites from 12 subjects) and control (36 sites from 12 subjects) sites or between the floras of the active and control sites and of 63 samples from 22 AP subjects that were examined previously in a cross-sectional study. By paired t test, no microbial species had a significantly greater association with active than with control sites. The only species that were detected in one or more samples from all subjects with active sites were Wolinella recta, Fusobacterium nucleatum, and Peptostreptococcus micros. Porphyromonas gingivalis and 9 other taxa were isolated from one-half or more of the persons with active sites. The composition of microbiological floras of all periodontitis samples was statistically significantly different from that of subjects with healthy gingiva. The composition of microfloras of sites in subjects with naturally-occurring gingivitis was intermediate between that of subjects with healthy gingiva and that of active and control sites in AP subjects.

Lactobacillus uli sp. nov. and Lactobacillus rimae sp. nov. are described. These organisms are short, gram-positive, strictly anaerobic, rod-shaped bacteria that have DNA G+C contents of 53 and 45 mol%, respectively, produce major amounts of lactic acid, and have been isolated from human gingival crevices and periodontal pockets. The major cellular fatty acid derivatives for both species are C18:1 cis-9 fatty acid methyl ester and C18:1 cis-9 dimethylacetyl. The type strain of L. uli is strain VPI D76D-27C (= ATCC 49627), and the type strain of L. rimae is strain D140H-11A (= ATCC 49626). Emended descriptions of Lactobacillus minutus (based on selected strains) and Streptococcus parvulus (based on many additional strains) also are given.

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DNAs of type strains and representative members of Actinomyces groups from the human periodontal flora and from other habitats were compared by using the S1 nuclease procedure to determine their genetic relatedness. One rather common group from the human periodontal flora, previously called "Actinomyces D08," is phenotypically distinct from, and genetically unrelated to, previously described species. We propose the name of Actinomyces georgiae for this organism; the type strain is strain ATCC 49285. Another common group from the human periodontal flora is Actinomyces israelii serotype II, which was found genetically distinct from the type strain of A. israelii (serotype I) and from other previously described species of Actinomyces. We propose the name Actinomyces gerencseriae for this organism; the type strain is strain ATCC 23860. A. naeslundii serotype I strains were distinct from the other strains studied. A separate genospecies which included strains of A. naeslundii serotypes II and III and A. viscosus serotype II was delineated. Strains of Actinomyces serotype WVA 963 constitute an additional distinct genospecies. Because there are no reliable phenotypic tests, other than serological analyses, to differentiate Actinomyces serotype WVA 963 and the two genospecies of A. naeslundii, no taxonomic changes are proposed for these three genospecies.

Journal of Periodontal ResearchVolume 24, Issue 3 p. 222-223 Good's L-statistic–Rebuttal W. E. C. Moore, Corresponding Author W. E. C. Moore Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorL. V. H. Moore, Corresponding Author L. V. H. Moore Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorR. M. Smibert, Corresponding Author R. M. Smibert Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorI. J. Good, Corresponding Author I. J. Good Department of Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorJ. A. Burmeister, Corresponding Author J. A. Burmeister Department of Periodontics, Virginia Commonwealth University, Richmond, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorK. G. Palcanis, Corresponding Author K. G. Palcanis College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Addresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorR. R. Ranney, Corresponding Author R. R. Ranney College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Addresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this author W. E. C. Moore, Corresponding Author W. E. C. Moore Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorL. V. H. Moore, Corresponding Author L. V. H. Moore Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorR. M. Smibert, Corresponding Author R. M. Smibert Department of Anaerobic Microbiology, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorI. J. Good, Corresponding Author I. J. Good Department of Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorJ. A. Burmeister, Corresponding Author J. A. Burmeister Department of Periodontics, Virginia Commonwealth University, Richmond, VAAddresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorK. G. Palcanis, Corresponding Author K. G. Palcanis College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Addresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this authorR. R. Ranney, Corresponding Author R. R. Ranney College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Addresses: Departments of Anaerobic Microbiology and Statistics, Virginia Polytechnic Institute and State University, Blacksburg, VA, Department of Periodontics, Virginia Commonwealth University, Richmond, VA, College of Dentistry, University of Alabama at Birmingham, Birmingham, AL, U.S.A.Search for more papers by this author First published: May 1989 https://doi.org/10.1111/j.1600-0765.1989.tb02010.xAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL References Morris EC, Kowalski CJ. On the use of Good's L-statistic in the anlaysis of bacteriologic samples. J Periodont Res 1988; 23: 345. Volume24, Issue3May 1989Pages 222-223 ReferencesRelatedInformation