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PubMed1995

The microbiota of osseointegrated implants in patients with a history of periodontal disease.

The purpose of this study was to determine the presence of suspected periodontal pathogens in the peri-implant microflora of osseointegrated implants exposed 3 and 6 months to the oral environment of patients previously treated for periodontal disease. Subgingival microbial samples were taken in the deepest residual pocket of each quadrant in 10 patients before placement of ITI implants and in 10 patients before abutment connection of Brånemark implants. The samples were cultured using continuous anaerobic techniques. 4 patients were positive for P. gingivalis, 13 for P. intermedia, 2 for A. actinomycetemcomitans, 16 for Fusobacterium, 7 for C. rectus and 12 showed microscopic evidence of spirochetes. After 3 and 6 months exposure of the implants to the oral environment, one sample was obtained from the peri-implant sulcus in each patient. P. gingivalis was found in 2 patients in the peri-implant samples after 3 and 6 months. 6 peri-implant samples were positive for P. intermedia after 3 months, 7 after 6 months. Fusobacterium was present in 13 peri-implant samples after 3, and in 12 samples after 6 months. 4 patients showed evidence of for C. rectus after 3 months, 2 after 6 months. Spirochetes were seen in 3 peri-implant samples after 3 and in 5 after 6 months. None of the implants were found to be colonized by A. actinomycetemcomitans, although the organism was detected on teeth in 1 individual. Similar distribution patterns were noted for ITI and Brånemark type implants.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1991

Multicenter evaluation of tetracycline fiber therapy. III. Microbiological response.

In a multicenter study of the effects of tetracycline (TC) fiber therapy, subgingival plaque samples were tested for 6 probable periodontal pathogens by DNA probe analysis. Levels of Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas (Bacteroides) gingivalis, Prevotella intermedia (Bacteroides intermedius), and Wolinella recta were quantitatively determined in samples taken at baseline, and immediately after TC fiber removal, control fiber removal, and scaling and root planing. At untreated sites, samples were taken at baseline and 10 d later. Specificity of the DNA probe method was evaluated by testing the hybridization to 83 reference cultures. Interaction of the F. nucleatum probe with Fusobacterium periodonticum, and of the W. recta probe with Wolinella curva were the only cross-hybridizations noted. Species were detected at an average sensitivity of 2.9 x 10(4) organisms per sample. Approximately 70% of sites were initially infected with P. gingivalis and F. nucleatum; 50% with P. intermedia and E. corrodens; infections with W. recta and A. actinomycetemcomitans were less common (36% and 11% respectively). The average numbers of organisms found in the plaque samples were highest for F. nucleatum, P. gingivalis, and P. intermedia (ca. 10(6)). E. corrodens, W. recta, and A. actinomycetemcomitans occurred at 10-fold lower levels. Bacterial numbers and proportions of species in subgingival sites from the five centers did not differ appreciably. Both TC fiber therapy and scaling decreased the number of sites infected with all the monitored species. The bacterial composition at untreated sites and at sites where control fibers were placed was not significantly altered. The percentage reduction of the number of sites with detectable infection varied with each species: from 86% with W. recta to approximately 40% with P. gingivalis. Significant reduction of pocket depth and bleeding occurred at TC fiber-treated sites infected with each of the species. Significant attachment level gain occurred only at sites initially infected with P. gingivalis and treated with TC fibers.

Actinobacillus↗

PubMed1996

Anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis detected by indirect immunofluorescence.

Aiming to assess the presence of selected anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis. 25 central and lateral upper incisors presenting with radiographic evidence of chronic apical periodontitis were studied. The pulp chamber was opened under aseptic conditions and samples of the root canal content were collected with sterile absorbent paper points, which were placed and dispersed in test tubes containing reduced transport medium RTT. Aliquots were dried on glass slides and stained by indirect immunofluorescence for detection of Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia. The results showed a positive indirect immunofluorescence reaction in 24 of the 25 samples. Fourteen were positive for the specie Actinomyces viscosus, 12 for Prevotella intermedia, 10 for Fusobacterium nucleatum and 4 for Porphyromonas gingivalis. A semiquantitative assay was easily implemented for assessment of degree of infection by the organisms in individual cases.

Actinomyces viscosus↗

PubMed2004

Relationship between volatile sulphide compounds concentration and oral bacteria species detection in the elderly.

AIM: To evaluate the relationship between oral bacteria species detection and volatile sulphide compounds (VSC) concentration in the elderly. DESIGN: Cross-sectional microbiological and clinical VSC examination of elderly. PARTICIPANTS: Sixty-seven elderly people aged 75, who were functionally independent and dentate. METHODS: VSC (H2S and CH3SH) concentrations in the mouth air of subjects were measured using portable gas chromatography. Oral bacteria samples were taken from dental plaque and identification of bacteria species was accomplished using standard methods. RESULTS: Fewer than 20% of subjects showed more than 10 ng/10 ml of H2S (severe odour level). The detection rate of P. melaninogenica was significantly higher in elderly people with more than 10 ng/10 ml (p=0.043) levels. Fusobacterium had a tendency to be found in those with more than 10 ng/10 ml, but a significant relation was not found. CONCLUSIONS: The results suggest that Fusobacterium and P. melaninogenica may be involved in the production of H2S in the oral cavity of elderly people.

Aged↗

PubMed1993

Erythromycin, clarithromycin, and azithromycin: use of frequency distribution curves, scattergrams, and regression analyses to compare in vitro activities and describe cross-resistance.

MICs of erythromycin, clarithromycin, and azithromycin for 852 recent clinical isolates were determined by broth microdilution methods. Frequency distribution curves, scattergrams, and regression analyses were used to compare in vitro activities and describe cross-resistance. Clarithromycin was the most active drug against Bacteroides spp. but the least active against Haemophilus influenzae. Azithromycin was most active against H. influenzae, Moraxella catarrhalis, Pasteurella multocida, and Fusobacterium spp. but the least active against Streptococcus spp. and Enterococcus spp. All three drugs had equivalent activities against Staphylococcus spp. and gram-positive anaerobes. None of the three drugs was particularly active against members of the family Enterobacteriaceae or nonfermentative gram-negative bacilli, although concentrations of 4 micrograms of azithromycin per ml inhibited some strains of the family Enterobacteriaceae (particularly Escherichia coli and Citrobacter diversus) and Acinetobacter baumannii. Although relative drug activities varied by organism, organisms relatively susceptible to one were relatively susceptible to all and organisms relatively resistant to one were relatively resistant to all; an exception was fusobacteria, which were usually susceptible only to azithromycin. Cross-susceptibility and cross-resistance were, therefore, the rule (except for Fusobacterium spp.), although the percentage of susceptible organisms could be varied considerably on the basis of the selection of breakpoints.

Azithromycin↗

PubMed1976

Anaerobic bacteria from the large intestine of mice.

Anaerobic bacteria from the colon of laboratory mice were enumerated and isolated using strict anaerobic techniques. Direct microscopic counts revealed 4.4 X 10(10) organisms in each gram (wet weight) of colon contents. Actual cultural counts averaged 3.2 X 10(10) organisms, which was 73% of the direct microscopic count. The tentatively identified genera were Bacteroides, Eubacterium, Fusobacterium, Lactobacillus, Peptostreptococcus, and Propionibacterium. Strains of Fusobacterium, Lactobacillus, Peptostreptococcus, and Propionibacterium were biochemically homogeneous. Strains of Bacteroides and Eubacterium, on the other hand, were biochemically heterogeneous and were subdivided into several distinct groups. The data indicate that many of the isolates are different from previously described species of the respective genera and may belong to new species.

Anaerobiosis↗

PubMed1970

Dehydrogenase patterns in the study of Bacteroidaceae.

Enzyme patterns were obtained by starch-gel electrophoresis of cell-free extracts from organisms in the family Bacteroidaceae. Esterases, phosphatases, and lactic and succinic dehydrogenases were detected but offered no possibility for classification purposes. Alpha-glycerophosphate dehydrogenase and 6-phosphogluconate dehydrogenase were not detected. Zymograms of malic, glutamic, and glucose-6-phosphate dehydrogenases separated Bacteroides species from Sphaerophorus and Fusobacterium species. Malic dehydrogenase and glucose-6-phosphate dehydrogenase were found in Bacteroides but not Sphaerophorus and Fusobacterium. These dehydrogenase zymograms placed three gram-negative, nonsporeforming anaerobic rod isolates with the Bacteroides species. A close correlation was found between the classification of Bacteroidaceae by zymogram analysis and a numerical taxonomy scheme previously published from this laboratory.

Bacteria↗

PubMed1970

Effect of bile and desoxycholate on gram-negative anaerobic bacteria.

The bile tests for characterizing gram-negative anaerobic bacilli were reevaluated in prereduced anaerobically sterilized peptone-yeast-glucose broth, in thioglycollate broth, and on blood agar plates. Blood agar plates were unsatisfactory. The combination of 20% bile with 0.1% desoxycholate inhibited Fusobacterium, Bacteroides melaninogenicus, and B. oralis and sometimes Sphaerophorus necrophorus, but not B. fragilis or other Sphaerophorus species studied. Ten per cent bile with 0.05% desoxycholate was less satisfactory. There was no significant difference between fresh and commercial powdered bile. Desoxycholate (0.1% in thioglycollate broth) inhibited B. fragilis, Fusobacterium, B. melaninogenicus, B. oralis, and S. necrophorus, but not S. varius or S. mortiferus/S. ridiculosus. The bile and desoxycholate tests are simple to perform and helpful for characterization and classification of gram-negative anaerobic bacilli.

Animals↗

PubMed1971

Rifampin-blood-agar as a selective medium for the isolation of certain anaerobic bacteria.

A selective medium which allows detection of relatively small numbers of Fusobacterium varium in fecal specimens is described. Blood-agar containing 50 mug of rifampin per ml inhibits the growth of many species of Bacteriodes and of F. fusi-forme/nucleatum but allows good growth of F. varium and most strains of F. mortiferum. Quantitative cultures of 11 fecal specimens were done on rifampin and other selective and nonselective media. F. varium was recovered in counts of 10(6) and 10(7) per gram from two specimens on rifampin only. A third specimen yielded 10(10)F. varium on several media, including rifampin. Some Eubacterium and Clostridium species also grew on rifampin, and these ordinarily were distinguished from the Fusobacterium by colony morphology. This medium is of value in fecal flora studies and should be useful with other kinds of specimens where mixtures of organisms are common.

Agar↗

PubMed1983

Lipid A in anaerobic bacteria.

The ability of lipid A preparations from strains of Bacteroides, Fusobacterium, and Veillonella to inhibit the lipid A-anti-lipid A reaction in an enzyme-linked immunosorbent assay was tested. Anti-lipid A serum was prepared with lipid A from Salmonella minnesota R595, and lipid A from Escherichia coli EH100 was used as control antigen. Preparations from three of four different species of Bacteroides were unable to inhibit the anti-lipid A activity, whereas lipid A preparations from Fusobacterium and Veillonella strains inhibited 50% of the activity at 1 to 141 micrograms. One of the Bacteroides strains, Bacteroides oralis, showed a very weak inhibiting activity at the highest concentration used. The results confirm that Bacteroides species have a unique lipopolysaccharide structure, in contrast to other anaerobic genera which have a lipopolysaccharide structure similar to that of the Enterobacteriaceae.

Anaerobiosis↗

PubMed1967

Antibiotic susceptibility patterns as aids in classification and characterization of gram-negative anaerobic bacilli.

Patterns of susceptibility of gram-negative anaerobic bacilli to antibiotics have been found to be distinctive and of significant help in classification and identification. Five major groups of gram-negative anaerobic bacilli have been defined on the basis of morphological and biochemical criteria. Antibiotic sensitivity patterns conform to these groupings and provide additional taxonomic criteria. The Bacteroides fragilis group is resistant to penicillin G, whereas the other groups are generally sensitive. B. fragilis strains are relatively sensitive to erythromycin, whereas the Sphaerophorus necrophorus group is resistant. B. melaninogenicus strains, B. oralis, and Fusobacterium are all more sensitive to kanamycin and neomycin than the other two groups. Kanamycin is more active against Fusobacterium strains than neomycin, but less active against all other groups. Colistin or polymyxin B is useful for distinguishing between the resistant B. fragilis and the sensitive S. necrophorus. Antibiotic susceptibility determinations may be more readily performed in clinical laboratories than certain biochemical tests recommended for differentiation of the gram-negative anaerobic bacilli and may serve as helpful adjuncts to morphological and biochemical observations in classifying and characterizing these organisms. The use of standardized procedures for antibiotic susceptibility tests is essential if comparable results are to be obtained in different laboratories.

Anti-Bacterial Agents↗

PubMed1979

Characterization of anaerobic gram-negative bacilli by using rapid slide tests for beta-lactamase production.

A total of 175 isolates of anaerobic gram-negative bacilli were tested for beta-lactamase production by using a slide test modification of the chromogenic cephalosporin (Nitrocefin, Glaxo, Middlesex, England) assay and the iodometric slide test. Included isolates were Bacteroides melaninogenicus (46), B. fragilis (78), other Bacteroides isolates (21), Fusobacterium (25), and other gram-negative bacilli (5). Both slide tests detected 25 B. melaninogenicus isolates that were beta-lactamase producers (minimal inhibitory concentration of penicillin was greater than 0.78 micrograms/ml). beta-Lactamase produced by the other gram-negative anaerobes could only be detected by the Nitrocefin assay. This assay was positive in 70 or 77 B. fragilis against which the minimal inhibitory concentration of penicillin was greater than 0.78 micrograms/ml. Ten of 11 other species of Bacteroides against which the minimal inhibitory concentration of penicillin was greater than 0.78 micrograms/ml were also Nitrocefin test positive. Minimal inhibitory concentrations of penicillin against all isolates of Fusobacterium and unidentifified gram-negative bacilli were less than or equal to 0.78 micrograms/ml and were Nitrocefin assay negative. beta-Lactamase-producing strains of B. melaninogenicus can be differentiated because both the slide iodometric and Nitrocefin assays will be positive, whereas beta-lactamase produced by other Bacteroides will only be detected by the Nitrocefin assay. Such penicillin-resistant isolates could be detected and reported to clinicians before final identification.

Bacteriological Techniques↗

PubMed1980

The effect of intensive antibacterial therapy on the sulcular environment in monkeys. Part I. Changes in the bacteriology of the gingival sulcus.

The changes induced in the bacteriology of the gingival sulcus were evaluated as part of a study considering the keratinizing potential of the sulcular epithelium when bacterial plaque was essentially eliminated. Two Rhesus monkeys were scaled and placed on a daily therapeutic regimen which included a prophylaxis, systemic tetracycline, and topical chlorhexidine. Over the 40 day experimental period and 74 days post-therapy, subgingival plaque samples were taken periodically. The plaque samples were cultured anaerobically and aerobically to determine the predominant bacterial flora. The total cultivable bacterial flora decreased from initial levels by greater than 99.9% with the antibacterial therapy. The flora shifted with therapy from one dominated by anaerobic organisms, including Bacteroides melaninogenicus (18%) and Fusobacterium species (13.9%), to a flora dominated by organisms growing aerobically. During treatment B. melaninogenicus and Fusobacterium species were not detected in any sample. After cessation of all therapy the anaerobes increased to dominance again, but B. melaninogenicus remained undetectable through 74 days post-therapy.

Actinomyces↗

PubMed1982

The bacteriology of acute necrotizing ulcerative gingivitis.

Plaque samples from 22 ulcerated sites in eight patients with ANUG were cultured using quantitative anaerobic procedures and were examined microscopically. The partial characterization of the predominant cultivable flora revealed a constant flora comprised of a limited number of bacterial types and a variable flora composed of a heterogeneous array of bacterial types. This constant flora would appear to be pathognomonic of acute necrotizing ulcerative gingivitis (ANUG) and included the various Treponema and Selenomonas sp., which comprised about 32 and 6%, respectively, of the microscopic count; B. melaninogenicus ssp. intermedius and Fusobacterium sp., which averaged 24 and 3%, respectively, of the viable count. One week of metronidazole treatment caused a prompt resolution of clinical symptoms, which coincided with a significant reduction in the plaque proportions of the Treponema sp., B. melaninogenicus ssp., intermedius and Fusobacterium sp. for at least 2 to 3 months following treatment. Thus, the same anaerobic species which were numerically associated with the ANUG lesion were also selectively reduced in the plaque flora following resolution of the infection. This supports a role for the above species in the ulcerative stage of the lesion but does not demonstrate that these specific anaerobes initiated the infection. although not confirmed by the data, it was proposed that these particular anaerobic species gained ascendency in the plaque as a result of being selected through the availability of host-derived nutrients in individuals who had undergone certain physiological and psychological stresses.

Bacteria↗

PubMed1984

Human neutrophil migration under agarose to bacteria associated with the development of gingivitis.

In clinically healthy gingiva and increasingly with the development of inflammation, neutrophils are found in the gingival tissues and sulcus. This study evaluated the relative ability of bacteria associated with gingival health and developing inflammation to stimulate this increase in neutrophil accumulation. Dialyzed bacterial sonic extracts (BE) in buffer and pooled human serum (PHS) from pure cultures of Streptococcus sanguis. Actinomyces viscosus, A naeslundii, Bacteroides intermedius, Fusobacterium sp and Veillonella sp were tested for stimulation of human neutrophil migration under agarose. In addition, fractions of S sanguis culture fluids (CFs) from Sephadex G-10 chromatography were evaluated. All BE solutions were incubated for 1 hour at 37 degrees C and heat-inactivated prior to testing. All BEs in buffer attracted neutrophils, with the greatest responses seen to S sanguis and B intermedius followed by A viscosus. Migration to all BEs in PHS was greater than in buffer, suggesting that all BEs are capable of generating serum chemoattractants. A viscosus BE activated serum attractants to the greatest degree. CFs of S sanguis, A viscosus, and to a lesser degree, Fusobacterium sp, also attracted neutrophils. Evidence from [3H]FMLP competitive ligand-binding assays indicated that S sanguis CFs contained low molecular weight (less than 700) chemoattractants, which were probably formylmethionyl oligopeptide-like materials. Of the bacteria associated with health, S sanguis and A viscosus appeared at least as able to generate chemoattractants during growth or with exposure to serum as bacteria associated with gingivitis. This observation suggests that these "healthy" bacteria, which are found in greater numbers with developing inflammation, may mediate increased neutrophil transmigration in early disease.

Actinomyces↗

PubMed1985

Effects of scaling and root planing on subgingival microbial proportions standardized in terms of their naturally occurring distribution.

This investigation included a cross-sectional component (284, 20-40-year-old subjects/568 sites) and a longitudinal component (19 subjects with moderate to advanced periodontitis/76 sites). Subgingival plaque samples and clinical data were obtained from all upper second bicuspids and first molars in the longitudinal subjects and from both upper first molars in the cross-sectional subjects. The purpose was to: determine/confirm changes in proportions of selected subgingival microorganisms following root planning combined with conventional oral hygiene procedures in the longitudinal subjects, evaluate those changes in terms of an estimate of the naturally occurring distribution of subgingival microorganisms based on data obtained from the cross-sectional subjects and relate shifts in the subgingival microorganisms to changes in clinical measurements. Changes following treatment at sites with pre-instrumentation probing depths greater than or equal to 4.5 mm included cocci, 18.3 to 46.9%; spirochetes, 20.9 to 3.1%; total motile organisms, 28.5 to 5.9%; Fusobacterium spp., 10.2 to 2.2%; and dark-pigmented Bacteroides spp., 14.0 to 9.7%. Translating proportions of microorganisms in samples from subjects in the longitudinal treatment study to equivalent percentile ranks within the large cross-sectional data base demonstrated that at probing depths greater than or equal to 4.5 mm cocci moved from below the 50th percentile in our estimate of the naturally occurring distribution of subgingival microorganisms to above, and spirochetes, total motile organisms and Fusobacterium spp. moved from above the 50th percentile to below. The microbiological findings were consistent with statistically significant improvements in clinical measurements. Statistically significant changes also occurred in microorganisms at sites with initial probing depths less than 4.5 mm.

Adult↗

PubMed1992

The effect of cyclosporin and lipopolysaccharide on fibroblasts: implications for cyclosporin-induced gingival overgrowth.

Drug-induced gingival overgrowth is an adverse side effect associated principally with 3 different types of drugs; specifically the antiepileptic phenytoin, the calcium channel antagonist nifedipine, and the immunosuppressant cyclosporin. The present study has analyzed the effect of cyclosporin and lipopolysaccharide on fibroblasts from 3 different sources: 1) normal healthy human gingiva (NHGF); 2) overgrown gingiva from 2 patients taking cyclosporin (CHGF); and 3) human fetal lung (WI-38). Fibroblasts isolated from cyclosporin-associated gingival overgrowth were significantly less responsive to cyclosporin in terms of DNA, total protein, and proteoglycan synthesis. This finding supports the in vivo response where few fibroblasts are seen but marked overgrowth of fibrous tissue occurs. Lipopolysaccharide derived from Fusobacterium nucleatum and Escherichia coli was capable of inhibiting DNA synthesis significantly in all 3 fibroblast types. Total protein synthesis by CHGF cells was inhibited differentially by Fusobacterium nucleatum LPS and addition of cyclosporin to this system resulted in reversal of the inhibition. A synergistic effect was noted when the proteoglycan output of NHGF cells was assessed in response to co-incubation with cyclosporin and Escherichia coli LPS. The study shows that bacterial LPS may be an important co-factor in the pathogenesis of cyclosporin-induced gingival overgrowth.

Cells, Cultured↗

PubMed1999

Effect of virginiamycin on ruminal fermentation in cattle during adaptation to a high concentrate diet and during an induced acidosis.

The objective of Exp. 1 was to compare the effects of virginiamycin (VM; 0, 175, or 250 mg x animal(-1) x d(-1)) and monensin/tylosin (MT; 250/ 90 mg x animal(-1) x d(-1)) on ruminal fermentation products and microbial populations in cattle during adaptation to an all-concentrate diet. Four ruminally cannulated, Holstein steers were used in a 4x4 Williams square design with 21-d periods. Steers were stepped up to an all-concentrate diet fed at 2.5% of BW once daily. Ruminal pH, protozoal counts, and NH3-N and VFA concentrations generally were unaffected by VM or MT. Mean counts of Lactobacillus and Streptococcus bovis were lower (P<.05) for VM-treated compared with control or MT-treated steers. Both VM and MT prevented the increase in Fusobacterium necrophorum counts associated with increasing intake of the high-concentrate diet observed in the control. The objective of Exp. 2 was to compare the effects of VM and MT on ruminal pH, L(+) lactate and VFA concentrations, and F. necrophorum numbers during carbohydrate overload. Six ruminally cannulated Holstein steers were assigned randomly to either the control, VM (175 mg/d), or MT (250 + 90 mg/d) treatments. Acidosis was induced with intraruminal administration of a slurry of ground corn and corn starch. The VM and MT premixes were added directly to the slurry before administration. Carbohydrate challenge induced acute ruminal acidosis (pH was 4.36 and L (+) lactate was 19.4 mM) in controls by 36 h. Compared with the controls, steers receiving VM or MT had higher (P<.05) ruminal pH, and the VM group had a lower (P<.05) L (+) lactate concentration. Fusobacterium necrophorum numbers initially increased in VM- and MT-administered steers. In the control steers, F. necrophorum was undetectable by 36 h. Virginiamycin seemed to control the growth of ruminal lactic acid-producing bacteria and, therefore, has the potential to moderate ruminal fermentation in situations that could lead to rapid production of lactic acid.

Acidosis↗