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PubMed1995

Microbiology of chronic suppurative otitis media in children in Surabaya, Indonesia.

The aerobic and anaerobic microbiology of 38 children from Surabaya, Indonesia, who suffered from chronic suppurative otitis media (CSOM) was studied using strict microbiological methodology. A total of 106 isolates (49 anaerobic and 57 aerobic) were recovered. Aerobic organisms alone were isolated from 11 (29%), anaerobic bacteria only in 4 (11%) and mixed aerobic and anaerobic flora were present in 23 (60%). The predominant organisms were Peptostreptococcus sp., Prevotella sp., Bacteroides sp., Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Fusobacterium sp. Thirty-nine beta-lactamase-producing bacteria were recovered from 22 (58%) patients. These findings demonstrate the role of penicillin resistant aerobic and anaerobic bacteria in the polymicrobial etiology of CSOM in children from Indonesia. Judicious use of antimicrobial therapy may prevent the development of antimicrobial resistance.

Adolescent↗
PubMed1991

Identification, and susceptibility to seven antimicrobial agents, of 61 gram-negative anaerobic rods from periodontal pockets.

Sixty-one cultures of Gram-negative anaerobic rods were isolated from deep periodontal pockets of patients with rapidly progressive periodontitis. Isolates were speciated as Bacteroides gingivalis (18 isolates), Bacteroides intermedius (8), Bacteroides oris (1), Bacteroides gracilis (17) and Fusobacterium nucleatum (17). Their susceptibilities, to seven antimicrobial agents, were determined in vitro using a plate dilution technique. Amoxycillin and amoxycillin with clavulanic acid were active against all isolates (MIC less than 1 mg/l) and proved the most effective agents tested. F. nucleatum and B. gracilis showed resistance to erythromycin; F. nucleatum had MIC values ranging from 0.03 mg/l up to 128 mg/l when tested with this, least effective agent. Metronidazole was effective against all isolates except for a few strains of B. gracilis (MIC less than 4 mg/l). Tetracycline hydrochloride and minocycline were active against all isolates except for a few strains of B. gracilis (MIC less than 2 mg/l with both minocycline and tetracycline hydrochloride). Penicillin proved less effective than amoxycillin with regard to inhibition of B. gracilis.

Amoxicillin↗
PubMed1991

Amoxycillin with clavulanic acid and tetracycline in periodontal therapy.

The effects of tetracycline and amoxycillin with clavulanic acid on the clinical parameters and subgingival flora of eight patients with rapidly progressive periodontitis was assessed. Subjects received either tetracycline 250 mg four times daily or amoxycillin 250 mg with clavulanic acid 125 mg three times daily for a period of 2 weeks together with subgingival scaling and root planning. Both treatment regimens produced significant reductions in bleeding on probing and probing pocket depths which were still present 16 weeks after the antibiotic therapy. A significant reduction in the mean percentage of black-pigmented Bacteroides spp., Fusobacterium nucleatum and anaerobic corroding bacilli was also obtained. Both treatment regimens were equally effective in reducing the clinical parameter and altering the subgingival flora. The MIC values for Bacteroides gingivalis (Porphyromonas gingivalis). Bacteroides intermedius (Prevotella intermedia) and F. nucleatum to amoxycillin with clavulanic acid remained constant throughout the period of investigation. The MIC values of these organisms to tetracycline increased.

Amoxicillin↗
PubMed1992

Synergistic degradation of bovine serum albumin by mutans streptococci and other dental plaque bacteria.

Mutans streptococci (Streptococcus mutans and Streptococcus sobrinus) exhibited low levels of proteolytic activity against the model protein substrate, FITC-labelled bovine serum albumin, when incubated alone. Inclusion of other members of the dental plaque microflora in the assay usually resulted in marked increases in the degree of proteolysis and a high level of synergy. Interactions between mutans streptococci and either Streptococcus oralis or Fusobacterium nucleatum gave rise to the greatest degree of synergistic proteolytic degradation.

Dental Plaque↗
PubMed1983

Jaw disease in macropod marsupials: bacterial flora isolated from lesions and from the mouths of affected animals.

Bacterial infections of the jaws are a common cause of death in macropods. Lesions and oral cavities from 50 affected animals yielded wide ranges of aerobic and anaerobic organisms. The most frequent isolate from lesions (81%) was Fusobacterium necrophorum, generally combined with other bacteria, but in 5 lesions, in pure culture. It was also isolated from 61% of mouths and this was the chief difference between the oral flora of affected and normal macropods. Other groups of organisms isolated from over 50% of lesions were: Gram-negative aerobic and anaerobic rods, streptococci, anaerobic Gram-positive cocci. Actinomycetes were isolated from 29% of lesions and from one lesion in pure culture. Differences in the flora were detected between lesions in bone and soft tissue and between closed and open lesions. Antibiotics were given to 22 animals, but without significant differences in frequencies of isolation of organisms between treated and untreated groups, and with no permanent elimination of infection. It was concluded that, while different organisms might be present in the complex of "jaw disease", the pathogenic agent in the majority of cases was F. necrophorum. Actinomycetes were capable of producing lesions in bone, but their role in "jaw disease" remains undefined.

Actinomycetales↗
PubMed1992

Comparative bacteriological studies on summer mastitis in grazing cattle and pyogenes mastitis in stabled cattle in Denmark.

A total of 143 secretions from clinical cases of summer mastitis (SM) in grazing cattle and from 89 cases of pyogenes mastitis (PM) in stabled cattle were examined bacteriologically. The typical bacteriological finding was a mixed flora in which the predominant organisms were Actinomyces pyogenes (SM-70%, PM-85%), Peptostreptococcus indolicus (54%, 54%), a microaerophilic coccus (Stuart-Schwan coccus) (26%, 25%), Fusobacterium necrophorum biovar B (22%, 12%), Bacteroides melaninogenicus (20%, 9%) and Streptococcus dysgalactiae (21%, 5%). All except six cases occurred in non-lactating animals or within three weeks after parturition. The majority of animals (about 90%) had only one quarter affected and no differences in quarter distribution were observed between the two groups.

Actinomyces↗
PubMed1983

A comparison of four methods for detecting beta-lactamase in anaerobic bacteria.

Four methods for detecting beta-lactamase activity in anaerobic bacteria were evaluated and compared to a nitrocefin saturated filter paper test. The methods studied were Cefinase, PADAC, Beta Lactam, and the slide iodometric technique. Fifty anaerobes were tested including species of Bacteroides, Fusobacterium, and Clostridium. The percentages of agreement with the nitrocefin test are: Cefinase 100%, PADAC 96%, slide iodometric 78%, and Beta Lactam 72%. Positive nitrocefin tests have been shown to correlate with a penicillin MIC greater than 0.78 micrograms/ml.

Bacteria, Anaerobic↗
PubMed1986

Comparative activity of SCH 34343 against anaerobic bacteria.

The in vitro activity of SCH 34343 was compared with that of imipenem, clindamycin, cefoxitin, and metronidazole against 78 strains of anaerobic bacteria in the presence and absence of blood. Wilkins-Chalgren agar was used. SCH 34343 and imipenem were the most active agents, inhibiting all strains at less than or equal to 8 micrograms/ml. The addition of blood had little effect on the activity of any of the agents. Seven strains were unable to grow on Wilkins-Chalgren agar. Even with the addition of blood, one strain each of Bacteroides asaccharolyticus and Bacteroides bivius and two strains of Fusobacterium were unable to grow. Sixty-eight of the strains were tested against SCH 34343 and imipenem on Brucella lysed blood agar. Minimal inhibitory concentrations tended to be somewhat higher on Brucella lysed blood agar than on Wilkins-Chalgren agar, and all strains were able to grow on Brucella lysed blood agar.

Anti-Bacterial Agents↗
PubMed1986

Activity of cefotetan against anaerobic bacteria.

The in vitro activity of cefotetan, a new cephamycin, was compared with that of cefoxitin, cefotaxime, moxalactam, and piperacillin against 368 strains of anaerobic bacteria. Cefotetan was similar to cefoxitin, moxalactam, and piperacillin in its activity against B. fragilis, B. vulgatus, Bacteroides species, anaerobic gram-positive cocci, Fusobacterium and Clostridium species. High resistance rates for cefotetan, however, were demonstrated among B. thetaiotaomicron-ovatus group and B. distasonis. Increasing inoculum concentrations had little effect on the activity of cefotetan.

Anti-Bacterial Agents↗
PubMed2005

The effects of intra-amniotic injection of periodontopathic lipopolysaccharides in sheep.

OBJECTIVE: Periodontal disease may cause several complications of pregnancy, including fetal death. The purpose of this study was to investigate in sheep the effects of the intra-amniotic injection of lipopolysaccharide from 3 periodontopathic organisms and to compare these effects with those resulting from similar injection of Escherichia coli lipopolysaccharide. The outcomes that were studied included the rates of fetal death and the features of inflammation and lung maturation in survivors. STUDY DESIGN: At 118 days of pregnancy, ewes that were bearing single fetuses were allocated at random to receive intra-amniotic injections of saline solution (n = 13 fetuses), or lipopolysaccharide from Porphyromonas gingivalis (in doses from 0.1 to 10 mg [n = 22 fetuses]), Actinobacillus actinomycetemcomitans (10 mg [n = 6 fetuses]; 1 mg [n = 6 fetuses]), Fusobacterium nucleatum (10 mg [n = 6 fetuses]) or Escherichia coli (10 mg [n = 14 fetuses]; 1 mg [n = 7 fetuses]). Surviving fetuses were delivered abdominally at 125 days of gestation (term, 150 days). RESULTS: When compared with Escherichia coli lipopolysaccharide at similar dosages, periodontopathic lipopolysaccharides had high rates of fetal lethality. Only 6 of 22 fetuses that were exposed to intra-amniotic Porphyromonas gingivalis lipopolysaccharide survived doses of 0.1 to 10 mg, and only 3 of 6 fetuses survived 10-mg Actinobacillus actinomycetemcomitans lipopolysaccharide. Escherichia coli lipopolysaccharide did not cause fetal loss when given at doses of 10 mg (n = 14 fetuses) or 1 mg (n = 7 fetuses). Fetuses that survived exposure to these lipopolysaccharides showed features of inflammation in amniotic fluid and cord blood at birth and enhanced lung maturation. CONCLUSION: Lipopolysaccharides from these 3 periodontopathic organisms have much higher rates of fetal lethality than Escherichia coli lipopolysaccharide but can cause similar intrauterine inflammatory responses and improvements in lung volumes in survivors. Sources of inflammation that are distant from the uterus may underlie a proportion of unexplained stillbirth and other complications of pregnancy.

Acid-Base Equilibrium↗
PubMed2005

Prevalence of aerobic and anaerobic uterine bacteria during peripartum period in normal and dystocia-affected buffaloes.

Parturition complications predispose establishment of uterine infections, which in turn affect subsequent fertility. The aim of present study was to characterize and compare the type of bacterial flora prevalent within the uterine lumen of dystocia-affected buffaloes and compare them with the normally calving buffaloes. The study was conducted on 40 buffaloes; of which 10 calved normally (Group I) and 30 were treated for dystocia (Group II). Bacteriological examination was performed using uterine swabs, which were collected before delivery, immediately after delivery and day's 24-60 postpartum. A total of 30 uterine swabs from Group I and 79 swabs from Group II were collected, of which 19 (63.3%) and 71 (89.9%) yielded significant bacterial growth, respectively. A total of 205 isolates belonging to 10 different genera of bacteria were identified, 8 facultative anaerobes and 2 obligate anaerobes. In Group II, 91.6% of the bacteria positive swabs (n = 71) yielded mixed cultures, whereas the remainder being pure cultures. In contrast, 89.5% of the bacteria positive swabs of Group I (n = 19) yielded pure cultures. Mixed infections comprised mostly Arcanobacter (Actinomyces) pyogenes together with obligate anaerobes, Fusobacterium spp. and Bacteroides spp. In Group II, the frequency of incidental and coliform group bacteria was highest at the time of parturition, i.e., before and immediately after delivery, and decreased to nil during the 24-60-day postpartum period. However, in Group I, the incidental and coliform group of bacteria present at the time of parturition apparently persisted beyond the period when uterine involution is complete. The frequency of obligate anaerobes and A. pyogenes at the time of parturition was nil in the Group I while they predominated in dystocia-affected buffaloes (Group II). During the postpartum period of 24-60 days, the frequency of both obligate anaerobes and A. pyogenes increased significantly in Group II, whereas in Group I, only occasional isolates were obtained. To conclude, at the time of calving the prevalence of obligate anaerobes and A. pyogenes occurring in combination was highest in dystocia-affected buffaloes, and further increased in the postpartum period suggesting that these infections act synergistically.

Actinomyces↗
PubMed2005

Multi-target antimicrobial actions of zinc against oral anaerobes.

OBJECTIVE: Zinc is used in oral care products as an antiplaque/antigingivitis agent. Our objective was to assess the antimicrobial actions of zinc against oral anaerobes associated with gingivitis, specifically Fusobacterium nucleatum and Prevotella intermedia, with focus on catabolism and oxidative metabolism. METHODS: The oral anaerobes were grown in complex medium in an anaerobic chamber, harvested by centrifugation and used directly for experiments with suspensions. Biofilm growth involved super-infection by F. nucleatum of an initial biofilm formed by Streptococcus sanguis. RESULTS: Zn(2+) inhibited catabolism of glutamate, glutamyl-glutamate, glucose and fructose by F. nucleatum cells in suspensions with ID(50) values, respectively, of 0.05, 0.005, 0.01 and 0.01 mM. The ID(50) value for inhibition of glutamate catabolism by biofilms was 0.10 mM. Inhibition of glutamate catabolism could be related to inhibition of substrate uptake and of 2-oxoglutarate reductase. Zn(2+) also inhibited catabolism of aspartate or aspartyl-aspartate by P. intermedia with ID(50) values of 0.07 and about 0.03 mM, respectively. Respiration of intact cells of F. nucleatum and NADH oxidase in cell extracts were sensitive to zinc with ID(50) values, respectively, of about 1.0 and 1.4 mM. Zinc also inhibited production of hydrogen peroxide by F. nucleatum (ID(50) = ca. 0.04 mM.) but at high concentrations acted to potentiate and enhance peroxide killing of the anaerobe. CONCLUSION: Zn(2+) is a potent inhibitor of catabolism by F. nucleatum and P. intermedia, including catabolism of peptides, which can be degraded to yield inflammatory metabolic end products. Zn(2+) also inhibits O(2) metabolism of F. nucleatum by about 50% and hydrogen peroxide production nearly completely but also enhances killing by peroxide added to cells.

Anaerobiosis↗
PubMed2005

Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR.

Periodontitis is a multi-factorial chronic inflammatory and destructive disease of the tooth-supporting tissues. Quantitative anaerobic culture techniques have been used for microbial diagnosis of the different forms of the disease. The aim of this study was to compare real-time PCR with quantitative anaerobic culture for detection and quantification of 5 prominent periodontal pathogens. Real-time PCR assays with the 16s rRNA genes of Actinobacillus actinomycetemcomitans, Prevotella intermedia, Tannerella forsythensis, Peptostreptococcus micros and Fusobacterium spp. were developed. The PCR was validated on pure cultures of various bacterial strains. Subsequently, subgingival plaque samples from 259 adult patients with periodontitis were analyzed with quantitative anaerobic culture and real-time PCR. A standard curve for DNA quantification was created for each primer-probe set based on colony-forming units equivalents. All bacterial species were correctly identified. The lower limits of detection by PCR varied between 1-50 colony-forming units equivalents depending on the species. No cross-reactivities with heterologous DNA of other bacterial species were observed. Real-time PCR results showed a high degree of agreement with anaerobic culture results. Real-time PCR is a reliable alternative for diagnostic quantitative anaerobic culture of subgingival plaque samples.

Adult↗
PubMed2004

Dehydration of (R)-2-hydroxyacyl-CoA to enoyl-CoA in the fermentation of alpha-amino acids by anaerobic bacteria.

Several clostridia and fusobacteria ferment alpha-amino acids via (R)-2-hydroxyacyl-CoA, which is dehydrated to enoyl-CoA by syn-elimination. This reaction is of great mechanistic interest, since the beta-hydrogen, to be eliminated as proton, is not activated (pK 40-50). A mechanism has been proposed, in which one high-energy electron acts as cofactor and transiently reduces the electrophilic thiol ester carbonyl to a nucleophilic ketyl radical anion. The 2-hydroxyacyl-CoA dehydratases are two-component systems composed of an extremely oxygen-sensitive component A, an activator, and component D, the actual dehydratase. Component A, a homodimer with one [4Fe-4S]cluster, transfers an electron to component D, a heterodimer with 1-2 [4Fe-4S]clusters and FMN, concomitant with hydrolysis of two ATP. From component D the electron is further transferred to the substrate, where it facilitates elimination of the hydroxyl group. In the resulting enoxyradical the beta-hydrogen is activated (pK14). After elimination the electron is handed-over to the next incoming substrate without further hydrolysis of ATP. The helix-cluster-helix architecture of component A forms an angle of 105 degrees, which probably opens to 180 degrees upon binding of ATP resembling an archer shooting arrows. Therefore we designated component A as 'Archerase'. Here, we describe 2-hydroxyglutaryl-CoA dehydratase from Acidaminococcus fermentans, Clostridium symbiosum and Fusobacterium nucleatum, 2-phenyllactate dehydratase from Clostridium sporogenes, 2-hydroxyisocaproyl-CoA dehydratase from Clostridium difficile, and lactyl-CoA dehydratase from Clostridium propionicum. A relative of the 2-hydroxyacyl-CoA dehydratases is benzoyl-CoA reductase from Thauera aromatica. Analogous but unrelated archerases are the iron proteins of nitrogenase and bacterial protochlorophyllide reductase. In anaerobic organisms, which do not oxidize 2-oxo acids, a second energy-driven electron transfer from NADH to ferredoxin, the electron donor of component A, has been established. The transfer is catalysed by a membrane-bound NADH-ferredoxin oxidoreductase driven by an electrochemical Na(+)-gradient. This enzyme is related to the Rnf proteins involved in Rhodobacter capsulatus nitrogen fixation.

Acidaminococcus↗
PubMed2005

DNA-DNA hybridization demonstrates multiple bacteria in osteoradionecrosis.

Bone necrosis secondary to radiation was previously attributed to trauma of devitalized bone and microbiological sepsis. However, conventional microbiological technique has failed to demonstrate microorganisms throughout osteoradionecrotic bone, claimed to be hypoxic, hypovascular and hypocellular. The aim of the present study was to examine such bone for bacteria using DNA-DNA hybridization. Compared to standard culture methods this technique enables the investigation of a vast number of bacteria in a fairly short time. Twelve deep medullary specimens from resected radionecrotic mandibles were studied. A multitude of bacterial species were detected, most of them anaerobic. Porphyromonas gingivalis was the most predominant organism, followed by Fusobacterium nucleatum subspecies polymorphum. All samples contained Actinomyces, Prevotella and F. nucleatum. The results of this study indicate that bacteria, particularly anaerobes, may play a more fundamental role in the pathophysiology of osteoradionecrosis than being merely surface contaminants.

Actinomyces↗
PubMed2005

Antibacterial action of photoactivated disinfection {PAD} used on endodontic bacteria in planktonic suspension and in artificial and human root canals.

OBJECTIVES: To measure antibacterial action of photoactivated disinfection (PAD) on endodontic bacteria in planktonic suspension and root canals. METHODS: Four bacteria, Fusobacterium nucleatum,Peptostreptococcus micros, Prevotella intermedia and Streptococcus intermedius, were tested in suspension. After mixing equal volumes of Tolonium chloride and bacterial suspension for 60s, each 200 microL of concentration (>10(6)cfu mL(-1)) was irradiated with light at 633+/-2 nm. Each energy dose/Tolonium chloride concentration combination was tested eight times, with controls. Prepared root canals in Training Blocs and extracted human teeth were inoculated with S. intermedius followed by 10 mg L(-1) Tolonium chloride or saline. Bacteria in canals were sampled before and after light irradiation. Student t-test assessed significance of changes in viable bacteria produced by treatment of either light or Tolonium chloride alone and light/Tolonium chloride combinations. RESULTS: In suspension, reductions in bacteria were highly significant (P<0.01) for light/Tolonium chloride combinations compared to light or Tolonium chloride alone. Maximum mean log reductions of 1.14 (P. intermedia), 2.48 (P. micros), 2.81 (F. nucleatum) and 6.73 (S. intermedius) were at 4.8 J/20 mg L(-1). Antibacterial action was increased by energy dose increase (not always significantly), but not by Tolonium chloride concentration. In control canals mean log reductions of 0.42 (Blocs) and 0.38 (teeth) from initial levels were not significant. PAD mean log reductions of 2.40 (Blocs) and 2.01 (teeth) were highly significant. Changes for PAD/energy dose combinations were not significant. CONCLUSION: PAD killed endodontic bacteria at statistically significant levels compared to controls. Kills varied with bacterial species.

Anti-Bacterial Agents↗
PubMed2006

Autoaggregation and coaggregation of bacteria associated with acute endodontic infections.

Biofilms and microbial aggregates are a common mechanism for the survival of bacteria in nature. Microbial aggregates have been associated with intraradicular and extraradicular endodontic disease. One objective of this study was to assess bacteria isolated from acute endodontic infections for autoaggregation and coaggregation. Another objective was to use both a conventional visual assay and a novel fluorescent dye-staining technique to study bacterial aggregation. Sixty-two strains of bacteria were isolated from 10 clinical samples of endodontic abscesses or cellulitis. Autoaggregation was detected in 35/62 (56.45%) of the bacteria using the visual assay. Coaggregation of bacteria from each of the samples was demonstrated for 29/183 (15.85%) bacterial pairs using the visual assay and 148/183 (80.87%) using the dye-staining assay. Coaggregation was observed for each of the 15 genera assayed, especially Prevotella, Streptococcus, and Fusobacterium. The dye-staining assay using a confocal microscope was a highly sensitive method to detect aggregation of bacteria.

Acute Disease↗
PubMed2006

Antimicrobial efficacy of medicated root canal filling materials.

Bacteria remaining within the root canal system are a significant factor in endodontic failures. The aim of this in vitro study was to evaluate the antimicrobial efficacy of commercially available gutta-percha formulations against common endodontic pathogens. Standard, tetracycline, or iodoform (also called medicated gutta-percha; MGP) containing gutta-percha were tested against Actinomyces israelii, A. naeslundii, Enterococcus faecalis, and Fusobacterium nucleatum using the agar diffusion method. Resilon points and tetracycline disks/E-tests were used as negative and positive controls, respectively. Resilon did not display antimicrobial properties. Standard gutta-percha and MGP inhibited F. nucleatum and A. naeslundii, with MGP also inhibiting A. israelii. Tetracycline containing gutta-percha was found to inhibit growth from all four bacterial species tested. The results of this study suggest that the use of tetracycline containing gutta-percha as an obturation material may be useful as a complement to the cleaning and disinfection phase of root canal procedures.

Actinomyces↗