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Oral Microbiology and Immunology Dec 1999Subgingival dental plaque consists mainly of microorganisms that derive their energy from amino acid fermentation. Their nutrient requirements are met by the subgingival...
Subgingival dental plaque consists mainly of microorganisms that derive their energy from amino acid fermentation. Their nutrient requirements are met by the subgingival proteolytic system, which includes proteases from microorganism and inflammatory cells, and substrate proteins from sulcus exudate, including albumin. To determine the selective effect of individual proteins on microbiota, we used albumin as the main substrate for growth. Eight subgingval plaque samples from untreated periodontal pockets of patients with adult periodontitis were inoculated in peptone yeast medium with bovine albumin (9 g/l). After three subculture steps, cell yields of the enrichment cultures at the medium with 0, 1.25, 2.5, 5, 10, and 20 g/l albumin were determined. Proteolytic activity (U/absorbance at 550 nm) of the enrichment cultures and different isolates derived from the cultures was estimated by the degradation of resorufin-labeled casein. It was observed that the yield of the mixed culture was albumin limited, and the proteolytic activities of the cultures in albumin broth were higher than in control (peptone broth). Among the isolates from the enrichment cultures, Peptostreptococcus micros, Prevotella melaninogenica, Prevotella buccae and Prevotella bivia demonstrated proteolysis. The frequent occurrence of Streptococcus gordonii and Streptococcus anginosus in the albumin cultures is explained by their ability to utilize arginine as an energy source for growth. Albumin in the medium was partly degraded by pure cultures but completely consumed in enrichment cultures, indicating synergy of bacterial proteinases. It is concluded that the subgingival microbiota possesses proteolytic activity and may use albumin as a substrate for their growth. Enrichment cultures on albumin may serve as a relatively simple in vitro model to evaluate the effects of proteinase inhibitors.
Topics: Bacterial Proteins; Caseins; Culture Media, Conditioned; Dental Plaque; Humans; Peptide Hydrolases; Peptostreptococcus; Periodontal Pocket; Prevotella; Protease Inhibitors; Serum Albumin, Bovine
PubMed: 10895689
DOI: 10.1034/j.1399-302x.1999.140603.x -
Antimicrobial Agents and Chemotherapy Sep 1999Gemifloxacin mesylate (SB 265805), a new fluoronaphthyridone, was tested against 359 recent clinical anaerobic isolates by the National Committee for Clinical Laboratory... (Comparative Study)
Comparative Study
Gemifloxacin mesylate (SB 265805), a new fluoronaphthyridone, was tested against 359 recent clinical anaerobic isolates by the National Committee for Clinical Laboratory Standards reference agar dilution method with supplemented brucella blood agar and an inoculum of 10(5) CFU/spot. Comparative antimicrobials tested included trovafloxacin, levofloxacin, grepafloxacin, sparfloxacin, sitafloxacin (DU-6859a), penicillin G, amoxicillin clavulanate, imipenem, cefoxitin, clindamycin, and metronidazole. The MIC(50) and MIC(90) (MICs at which 50 and 90% of the isolates were inhibited) of gemifloxacin against various organisms (with the number of strains tested in parentheses) were as follows (in micrograms per milliliter): for Bacteroides fragilis (28), 0.5 and 2; for Bacteroides thetaiotaomicron (24), 1 and 16; for Bacteroides caccae (12), 1 and 16; for Bacteroides distasonis (12), 8 and >16; for Bacteroides ovatus (12), 4 and >16; for Bacteroides stercoris (12), 0.5 and 0.5; for Bacteroides uniformis (12), 1 and 4; for Bacteroides vulgatus (11), 4 and 4; for Clostridium clostridioforme (15), 0.5 and 0.5; for Clostridium difficile (15), 1 and >16; for Clostridium innocuum (13), 0.125 and 2; for Clostridium perfringens (13), 0.06 and 0.06; for Clostridium ramosum (14), 0.25 and 8; for Fusobacterium nucleatum (12), 0.125 and 0.25; for Fusobacterium necrophorum (11), 0.25 and 0.5; for Fusobacterium varium (13), 0.5 and 1; for Fusobacterium spp. (12), 1 and 2; for Peptostreptococcus anaerobius (13), 0.06 and 0.06; for Peptostreptococcus asaccharolyticus (13), 0.125 and 0.125; for Peptostreptococcus magnus (14), 0.03 and 0.03; for Peptostreptococcus micros (12), 0.06 and 0.06; for Peptostreptococcus prevotii (14), 0.06 and 0.25; for Porphyromonas asaccharolytica (11), 0.125 and 0.125; for Prevotella bivia (10), 8 and 16; for Prevotella buccae (10), 2 and 2; for Prevotella intermedia (10), 0.5 and 0.5; and for Prevotella melaninogenica (11), 1 and 1. Gemifloxacin mesylate (SB 265805) was 1 to 4 dilutions more active than trovafloxacin against fusobacteria and peptostreptococci, and the two drugs were equivalent against clostridia and P. asaccharolytica. Gemifloxacin was equivalent to sitafloxacin (DU 6859a) against peptostreptococci, C. perfringens, and C. ramosum, and sitafloxacin was 2 to 3 dilutions more active against fusobacteria. Sparfloxacin, grepafloxacin, and levofloxacin were generally less active than gemifloxacin against all anaerobes.
Topics: Anti-Bacterial Agents; Anti-Infective Agents; Bacteria, Anaerobic; Fluoroquinolones; Gemifloxacin; Microbial Sensitivity Tests; Naphthyridines; Therapeutic Equivalency
PubMed: 10471570
DOI: 10.1128/AAC.43.9.2231 -
European Journal of Vascular and... Feb 1998
Topics: Aged; Aneurysm, False; Bacteremia; Bacteroidaceae Infections; Fatal Outcome; Humans; Male; Prevotella; Renal Artery Obstruction
PubMed: 9551059
DOI: 10.1016/s1078-5884(98)80141-x -
Oral Microbiology and Immunology Oct 1997This study aimed to evaluate the microbiota of necrotic pulp in teeth without carious lesions where the crown and root were intact and to test the sensitivity of this...
This study aimed to evaluate the microbiota of necrotic pulp in teeth without carious lesions where the crown and root were intact and to test the sensitivity of this microbiota to antibiotics in order to improve treatment. The necrotic pulp was sampled from 26 single-rooted teeth in intact pulp chambers. A total of 84 strains were isolated. The number of species isolated per tooth varied from 2 to 8, with a strong component (81%) of anaerobic bacteria. The most commonly represented species were Bacteroides gracilis, Propionibacterium acnes, Fusobacterium nucleatum, Prevotella buccae and Eubacterium lentum. The sensitivity of these organisms to amoxicillin, amoxicillin combined with clavulanate and tetracycline was evaluated by Etest on 38 isolates. For all strains tested, the minimum inhibitory concentration values obtained were low and substantially below effective serum concentrations for these antibiotics. These data enable us to devise suitable treatments for acute development of apical lesions and to prevent dissemination of this source of infection to the rest of the host.
Topics: Amoxicillin; Amoxicillin-Potassium Clavulanate Combination; Anti-Bacterial Agents; Bacteria; Bacteria, Anaerobic; Dental Pulp Cavity; Dental Pulp Necrosis; Drug Resistance, Microbial; Humans; Microbial Sensitivity Tests; Penicillins; Tetracycline
PubMed: 9467386
DOI: 10.1111/j.1399-302x.1997.tb00397.x -
Clinical Infectious Diseases : An... Sep 1997We characterized 52 anaerobic, gram-negative, nonpigmented, saccharolytic rods that were isolated from healthy and diseased subgingival sites of 16 family-owned dogs...
We characterized 52 anaerobic, gram-negative, nonpigmented, saccharolytic rods that were isolated from healthy and diseased subgingival sites of 16 family-owned dogs with spontaneous, clinically diagnosed periodontitis. Phenotypic features were determined with use of standard biochemical methods, by enzymatic profiling with the API ZYM system, and by cellular fatty acid profiling. Genotypic characterization was performed by DNA-DNA hybridization. Four phenotypic groups, defined as the Bacteroides fragilis group, the Prevotella buccae-like rods, the Prevotella heparinolytica/Prevotella zoogleoformans-like rods, and the slimy bile-tolerant rods, designated the Bacteroides pyogenes/Bacteroides tectum group, were detected. P. buccae and the B. pyogenes/B. tectum group organisms were isolated significantly more often (P values, < .05 and < .005) from the diseased than the healthy subgingival sites. The phenotypically similar group of bile-tolerant organisms, including B. pyogenes and B. tectum, most likely constitutes a major component of the anaerobic, gram-negative, saccharolytic microflora in periodontal lesions in dogs, a flora different from that in humans.
Topics: Animals; DNA, Bacterial; Dogs; Fatty Acids; Gingiva; Gram-Negative Anaerobic Bacteria
PubMed: 9310643
DOI: 10.1086/516209 -
Journal of Clinical Periodontology Aug 1997In 23 untreated adult periodontitis patients, the occurrence of beta-lactamase producing periodontal bacteria was determined. In addition to non-selective isolation... (Comparative Study)
Comparative Study
In 23 untreated adult periodontitis patients, the occurrence of beta-lactamase producing periodontal bacteria was determined. In addition to non-selective isolation media, selective isolation and growth of beta-lactamase positive subgingival bacterial species was carried out on blood agar plates supplemented with amoxicillin and plates with amoxicillin+clavulanic acid. Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Peptostreptococcus micros, Fusobacterium nucleatum, Bacteroides forsythus and Campylobacter rectus isolates from the non-selective medium were tested for beta-lactamase activity by a nitrocefin disk method (DrySlide) and by a laboratory chromogenic nitrocefin-based test. Isolates from the amoxicillin plates that were absent on the amoxicillin/clavulanic acid plates were identified and tested for beta-lactamase production. Based on the non-selective plates, six of 23 P. intermedia isolates, 2 of 19 B. forsythus isolates and 3 of 23 F. nucleatum isolates were beta-lactamase positive. The beta-lactamase positive species Prevotella loescheii, Prevotella buccae, Prevotella buccalis and Actinomyces spp were recovered from the selective amoxicillin plates. beta-Lactamase positive subgingival species were recovered from 17 of 23 patients (74%) but usually comprised low proportions of the subgingival microbiota (range < 0.01-15%). Comparison of the DrySlide test and the nitrocefin-based laboratory test revealed full agreement of test results. beta-Lactamase activity in whole subgingival plaque was detected in 12 patient samples (52%). It was concluded that beta-lactamase activity in subgingival bacteria in adult periodontitis is a common feature. However, since the majority of the samples showed only low-level enzymatic activity, the clinical relevance of this observation with regard to therapy with unprotected enzyme-susceptible beta-lactams is uncertain, though failure on the other hand, is difficult to rule out when a mechanism of resistance is present. The majority of beta-lactamase positive strains was found among species of the Prevotella genus.
Topics: Actinomyces; Adult; Aggregatibacter actinomycetemcomitans; Amoxicillin; Amoxicillin-Potassium Clavulanate Combination; Anti-Bacterial Agents; Bacteria; Bacteroides; Campylobacter; Cephalosporins; Chromogenic Compounds; Clavulanic Acids; Culture Media; Dental Plaque; Fusobacterium nucleatum; Gingiva; Humans; Indicators and Reagents; Penicillins; Peptostreptococcus; Periodontitis; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; beta-Lactamases
PubMed: 9266340
DOI: 10.1111/j.1600-051x.1997.tb00226.x -
Clinical and Diagnostic Laboratory... Jul 1997Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1...
Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region into intact Fab and Fc fragments. To investigate whether these enzymes are subject to inhibition in vivo in humans, we tested 34 sera from periodontally diseased and healthy individuals in an enzyme-linked immunosorbent assay for the presence and titers of inhibition of seven Prevotella and Capnocytophaga proteases. All or nearly all of the sera inhibited the IgA1 protease activity of Prevotella buccae, Prevotella oris, and Prevotella loescheii. A minor proportion of the sera inhibited Prevotella buccalis, Prevotella denticola, and Prevotella melaninogenica IgA1 proteases, while no sera inhibited Capnocytophaga ochracea IgA1 protease. All inhibition titers were low, ranging from 5 to 55, with titer being defined as the reciprocal of the dilution of serum causing 50% inhibition of one defined unit of protease activity. No correlation between periodontal disease status and the presence, absence, or titer of inhibition was observed. The nature of the low titers of inhibition in all sera of the IgA1 proteases of P. buccae, P. oris, and P. loescheii was further examined. In size exclusion chromatography, inhibitory activity corresponded to the peak volume of IgA. Additional inhibition of the P. oris IgA1 protease was found in fractions containing both IgA and IgG. Purification of the IgG fractions of five sera by passage of the sera on a protein G column resulted in recovery of inhibitory IgG antibodies against all three IgA1 proteases, with the highest titer being for the P. oris enzyme. These finding indicate that inhibitory activity is associated with enzyme-neutralizing antibodies.
Topics: Adolescent; Adult; Aged; Antibodies, Bacterial; Binding, Competitive; Capnocytophaga; Humans; Immunoglobulin A; Immunoglobulin G; Middle Aged; Periodontal Diseases; Prevotella; Serine Endopeptidases; Serine Proteinase Inhibitors
PubMed: 9220164
DOI: 10.1128/cdli.4.4.458-464.1997 -
Anaerobe 1997A case of a patient with a 20-year history of chronic otitis media complicated by cholesteatoma and brain abscess is described. A CT scan with contrast material showed...
A case of a patient with a 20-year history of chronic otitis media complicated by cholesteatoma and brain abscess is described. A CT scan with contrast material showed three abscess cavities in the right cerebellar hemisphere. A culture from a specimen of the cholesteatoma yielded a significant amount of growth of Bilophila wadsworthia, Bacteroides fragilis and Prevotella oris and a moderate growth of alpha-streptococci and Staphylococcus simulans. From the pus of the brain abscess we also isolated numerous Bilophila wadsworthia, Bacteroides fragilis and Prevotella oris and some Prevotella buccae and Peptostreptococcus anaerobius. No aerobes were present. The patient underwent a craniotomy and the biggest abscess was removed together with the capsule. The antimicrobial therapy included penicillin plus metronidazole and later augmentin. The result of the treatment was a complete cure and total recovery of the patient. This is the first documentation of isolation of B. wadsworthia in chronic otitis media and in brain abscess.
PubMed: 16887572
DOI: 10.1006/anae.1997.0084 -
The International Journal of Oral &... 1997The objective of this study was to examine inflammatory tissue in deep peri-implant bone pockets (> 5 mm) for anaerobic bacteria colonization. The peri-implant...
The objective of this study was to examine inflammatory tissue in deep peri-implant bone pockets (> 5 mm) for anaerobic bacteria colonization. The peri-implant inflammatory tissue of bone defects from 12 edentulous patients with 18 unsuccessful implants (IMZ type) was removed after surgical opening of the defects. After grinding the tissue with glass beads in nutrient solution, an aliquot of the suspension was plated and incubated on appropriate culture media. The quantitative and qualitative distribution of bacteria as a function of the tissue dry weight was determined (cell count/mg dry weight). The mean total cell count was 67 x 10(3) cells/mg dry weight. The following bacteria dominated: species of the family Bacteroidaceae (Prevotella intermedia, Prevotella buccae, Prevotella oralis, Prevotella melaninogenica, Prevotella denticola); Actinobacillus actinomycetemcomitans; Fusobacterium nucleatum; Capnocytophaga spp; and Eikenella corrodens. Bacteroidaceae and Actinobacillus actinomycetemcomitans were found particularly frequently. The increased colonization of these bacteria in deep peri-implant bone pockets is consistent with the currently held view of advanced periodontal lesions, whereby certain pathogens grow at a disproportionate rate in comparison with the total bacteria count under specific circumstances.
Topics: Aged; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Bacteria, Anaerobic; Bacterial Infections; Bacteroidaceae; Colony Count, Microbial; Dental Implantation, Endosseous; Dental Implants; Eikenella corrodens; Female; Fusobacterium nucleatum; Humans; Male; Periodontal Pocket; Prosthesis-Related Infections
PubMed: 9048462
DOI: No ID Found -
International Endodontic Journal Mar 1996Significant associations have been reported between (a) specific bacterial species isolated from root canals and (b) between individual bacterial species and endodontic...
Significant associations have been reported between (a) specific bacterial species isolated from root canals and (b) between individual bacterial species and endodontic symptoms and signs. The prime objective of this study was to determine whether particular combinations of specific bacteria are associated with individual endodontic symptoms and signs. Seventy root canals were investigated microbiologically taking care to maintain the viability of obligate anaerobes, which accounted for 64% of the total species isolated, including Peptostreptococcus micros, Prevotella melaninogenica, Prevotella oralis, Eubacterium aerofaciens, Eubacterium lentum, Fusobacterium nucleatum, Prevotella buccae and Prevotella intermedia. Significant associations were found between individual clinical features and the following pairs of species: (a) pain (37 cases) and Peptostreptococcus spp./Prevotella spp., Peptostreptococcus spp./Prevotella melaninogenica, Pstr. micros/Prev. melaninogenica (all P < 0.01); (b) swelling (23 cases) and Pstr. micros/Prevotella spp. (P < 0.01); (c) 'Wet' canal (57 cases) and Prevotella spp./Eubacterium spp. (P < 0.01), Peptostreptococcus spp./Eubacterium spp. (P < 0.05). Thus data from this investigation suggests that statistically significant associations exist between individual endodontic symptoms and signs and particular combinations of specific bacteria.
Topics: Adolescent; Adult; Aged; Bacteria, Anaerobic; Chi-Square Distribution; Child; Dental Pulp Cavity; Dental Pulp Diseases; Ecosystem; Eubacterium; Female; Fusobacterium; Humans; Male; Middle Aged; Peptostreptococcus; Prevotella; Streptococcus; Superinfection; Suppuration
PubMed: 9206427
DOI: 10.1111/j.1365-2591.1996.tb01164.x