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PubMed1992

Sensitization of oral bacteria to killing by low-power laser radiation.

Twenty-seven compounds were screened for their ability to sensitize Streptococcus sanguis to killing by light from a 7.3-mW Helium/Neon (HeNe) laser. Bacteria were mixed with various concentrations of the test compounds, spread over the surfaces of agar plates, and then exposed to light from the HeNe laser for various time periods. The plates were then incubated and examined for zones of inhibition. Those compounds found to be effective photosensitizers were then tested against Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Fusobacterium nucleatum. Toluidine blue O, azure B chloride, and methylene blue at concentrations of 0.005% (wt/vol) were effective photosensitizers of all four species, enabling killing of bacteria following exposure to laser light for only 30 s.

Aggregatibacter actinomycetemcomitans↗

PubMed1980

Aspects of anaerobic infections in animals.

The recent literature on clostridial infections of animals and infections due to gram-negative anaerobic rods is reviewed. It is stressed that quite recently the literature is concerned with animal models for the evaluation and study of the chemotherapy, immunity mechanisms and pathogenesis of Fusobacterium and Bacteroides infections in animals came into being.

Anaerobiosis↗

PubMed1986

Emergence of beta-lactamase producing anaerobic bacteria in the tonsils during penicillin treatment.

The emergence of beta-lactamase producing bacteria in the microflora in the oropharyngeal cavity was studied in ten healthy volunteers treated with 1 g phenoxymethylpenicillin b.i.d. for ten days. Beta-lactamase activity in saliva was also investigated. A significant increase in the number of beta-lactamase producing strains of Bacteroides species and Fusobacterium nucleatum was observed. One beta-lactamase producing Staphylococcus aureus strain was recovered in one of the volunteers before the penicillin administration started and three Staphylococcus aureus strains produced beta-lactamase after ten days of antibiotic treatment. Beta-lactamase-production in Haemophilus influenzae, Haemophilus parainfluenzae or Branhamella catarrhalis was not observed before, during or after the antibiotic treatment. Beta-lactamase activity was noted in the broth cultures from one volunteer colonized with a beta-lactamase producing Escherichia coli strain. Beta-lactamase activity in saliva was observed in all volunteers, the activity increasing significantly in parallel to the increase of beta-lactamase producing bacterial strains. Beta-lactamase activity in saliva was completely inhibited in vitro by clavulanic acid and p-chloromercurbenzoate and about 70-80 per cent of the activity was inhibited by cefoxitin. The increase of beta-lactamase producing bacteria in the oropharynx as a consequence of penicillin treatment raises doubt as to whether penicillin is the drug of choice in the treatment of tonsillitis caused by group A streptococci when previous treatment has failed.

Adult↗

PubMed2003

Effect of Porphyromonas gingivalis vesicles on coaggregation of Staphylococcus aureus to oral microorganisms.

Vesicles from the outer membrane of Porphyromonas gingivalis have the ability to aggregate a wide range of Streptococcus spp., Fusobacterium nucleatum, Actinomyces naeslundii, and Actinomyces viscosus. We found that in the presence of P. gingivalis vesicles, Staphylococcus aureus coaggregated with Streptococcus spp., and the mycelium-type Candida albicans, but not the yeast type. Autoaggregation of S. aureus in the presence of P. gingivalis vesicles is inhibited by L-arginine, L-lysine, and L-cysteine. Both the methicillin-sensitive (MSSA) and -resistant (MRSA) strains of S. aureus were able to coaggregate with Streptococcus spp., A. naeslundii, and A. viscosus when they were treated with P. gingivalis vesicles. P. gingivalis vesicle-treated mycelium-type C. albicans coaggregated with S. aureus, but the yeast-type did not. These results indicate that strains of S. aureus, including MRSA, could adhere to oral biofilms in dental plaque on the tooth surface or in the gingival crevice when P. gingivalis is present.

Actinomyces↗

PubMed2001

Antimicrobial activity of four root canal sealers against endodontic pathogens.

The antibacterial effects of various types of widely used endodontic sealers have not been compared systematically on facultative or obligate anaerobic endodontic pathogens. The aim of this study was to evaluate the antimicrobial properties of four commonly used endodontic sealers: two epoxy-resin-based sealers (AH26, AH plus), one zinc-oxide eugenol-based sealer (N2), and one calcium hydroxide-based sealer (Sealapex). The testing microbes were four facultative anaerobic species (Streptococcus mutans, Streptococcus sanguis, Escherichia coli, and Staphylococcus aureus) and four obligate anaerobic species (Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum, and Prevotella intermedia). The freshly mixed sealers were placed into the prepared wells of agar plates inoculated with the test microorganisms. After varying periods of incubation (2 days for facultative anaerobic species and 7 days for obligate anaerobic species), the zones of growth inhibition were observed and measured. All the sealers were distinctly different from each other in their antimicrobial activity. The sealers showed different inhibitory effects depending on the types and bacterial strains. N2 containing formaldehyde and eugenol proved to be the most effective against the microorganisms. The extreme antimicrobial potency of this root canal sealer must be weighted against its pronounced tissue toxic effect.

Analysis of Variance↗

PubMed2003

Intracellular production and extracellular release of oxygen radicals by PMNs and oxidative stress on PMNs during phagocytosis of periodontopathic bacteria.

In this study we investigated intracellular and extracellular oxygen radical production by polymorphonuclear leukocytes (PMNs) during the phagocytosis of periodontopathic bacteria. In in vitro assays, bacteria of the species Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Fusobacterium nucleatum were phagocytosed at 37 degrees C for 4 h by purified peripheral human PMNs from healthy subjects (n = 6). Superoxide production during phagocytosis was determined by flow cytometry and with a fluorescence/luminescence microplate reader. After phagocytosis, oxidative stress was determined by flow cytometry. Both the intracellular and extracellular oxygen radical production by PMNs phagocytosing F. nucleatum was significantly greater than that of PMNs phagocytosing P. gingivalis and A. actinomycetemcomitans ( P < 0.01 by the Mann-Whitney test). Moreover, after 4 h of incubation, the oxidative stress of PMNs phagocytosing F. nucleatum was significantly greater than that of PMNs phagocytosing P. gingivalis and A. actinomycetemcomitans. We conclude that a high level of superoxide production by PMNs may damage not only periodontopathic bacteria but also PMNs themselves, and may be correlated with the destruction of periodontal tissue.

Aggregatibacter actinomycetemcomitans↗

PubMed2005

Chronic sinusitis in children and adults: role of bacteria and antimicrobial management.

The nasopharynx serves as the reservoir for anaerobic bacteria as well as pathogenic bacteria that can cause respiratory infections including sinusitis. Some of these organisms possess the ability to interfere with the growth of potential pathogens and may play a role in preventing infections. Anaerobic bacteria emerge as pathogens as the infection becomes chronic. This may result from the selective pressure of antimicrobial agents that enable resistant anaerobic organisms to survive, and from the development of conditions appropriate for anaerobic growth, which include the reduction in oxygen tension and an increase in acidity within the sinus. Anaerobes were identified in chronic sinusitis in adults and children whenever techniques for their cultivation were employed. The predominant isolates were pigmented Prevotella, Fusobacterium, and Peptostreptococcus spp. The choice of antimicrobial agent in chronic sinusitis should provide coverage for the usual pathogens in acute sinusitis as well as beta-lactamase-producing aerobic and anaerobic organisms.

Adult↗

PubMed1992

Infection and labor. VII. Microbial invasion of the amniotic cavity in spontaneous rupture of membranes at term.

OBJECTIVE: The purpose of this study was to determine the frequency, microbiologic characteristics, and clinical significance of microbial invasion of the amniotic cavity in women with premature rupture of membranes at term. STUDY DESIGN: Amniocentesis was performed in 32 women with term premature rupture of membranes and amniotic fluid cultured for aerobic and anaerobic bacteria and Mycoplasmas. RESULTS: The prevalence of positive amniotic fluid cultures was 34.3% (11/32). The most common isolates were Ureaplasma urealyticum, Peptostreptococcus sp., Lactobacillus sp., Bacteroides fragilis, and Fusobacterium sp. Clinical chorioamnionitis occurred only in one patient with a positive amniotic fluid culture. Her neonate had ophthalmitis. Three patients (9.4%) had endometritis. Among women who were delivered vaginally, those with a positive amniotic fluid culture had a significantly higher rate of endometritis than those with a negative culture (33% [3/9] vs 0% [0/20], respectively, p = 0.023). CONCLUSIONS: These data indicate that microbial invasion of the amniotic cavity occurs in approximately one third of patients with preterm premature rupture of membranes. Microbial invasion of the amniotic cavity is a risk factor for endometritis in women with term premature rupture of membranes.

Amnion↗

PubMed1987

Bacterial adherence on replicas of sodium dodecyl sulfate-polyacrylamide gels.

A method for determining which molecules in a complex mixture of proteins can function as bacterial receptors was devised. Salivary proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose. Bacteria that were metabolically labeled with 3H or externally labeled with 125I were incubated on the nitrocellulose replicas. After 18 h at 4 degrees C, the unbound cells were removed by repeated washing of the replicas, and the bands to which the radiolabeled bacteria bound were visualized by autoradiography. By this technique, Fusobacterium nucleatum, which adheres via carbohydrate residues on receptor molecules, and Staphylococcus aureus, which recognizes the peptide portion of fibronectin, were shown to bind specifically to their respective receptors. These results suggest that this method can be useful for profiling bacterial binding to either the carbohydrate or the protein portions of molecules present in complex mixtures, such as those composing biological fluids or tissue substrates. Structural specificities, such as recognition sequences formed by certain oligosaccharides, could be further investigated by adding the appropriate simple sugars, as well as oligosaccharide inhibitors, to the incubation medium. The latter approach is particularly important since most glycoproteins carry multiple N- and O-linked carbohydrate substituents that could serve as bacterial receptors.

Bacterial Adhesion↗

PubMed1984

The effect of storage in liquid nitrogen on the recovery of human dental plaque bacteria.

Centrifuged deposits of various individual bacteria from type-culture collections and dental plaque samples were immersed in liquid nitrogen for periods of up to 2 years. All the organisms were recovered after storage in liquid nitrogen but Fusobacterium nucleatum (ATCC 10953) and Campylobacter fetus (NCTC 10354) consistently showed a reduction in the number of colony-forming units after freezing. The main effect on bacterial proportions in plaque samples was a reduction in aerobic Gram-negative cocci. Thus, storage in liquid nitrogen may be useful for comparative studies of dental plaque and as a means of storing individual organisms.

Adolescent↗

PubMed1992

Sensitization of oral bacteria in biofilms to killing by light from a low-power laser.

Biofilms of Streptococcus sanguis, Porphyromonas gingivalis, Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans were prepared on the surfaces of agar plates and a number of compounds were screened for their ability to sensitize bacteria in these biofilms to killing by light from a 7.3 mW Helium/Neon (He/Ne) laser. Toluidine blue O and methylene blue enabled detectable killing of all four target organisms after exposure to He/Ne light for 30 s. Aluminium disulphonated phthalocyanine, haematoporphyrin HCl and haematoporphyrin ester were effective photosensitizers of only some of the target organisms. These findings suggest that lethal photosensitization may be an effective means of eliminating periodontopathogenic bacteria from dental plaque.

Aggregatibacter actinomycetemcomitans↗

PubMed1992

Correlation of haemagglutination activity with trypsin-like protease activity of Porphyromonas gingivalis.

Porphyromonas gingivalis is a Gram-negative anaerobic bacterium associated with various forms of periodontal disease. Several characteristics of P. gingivalis are thought to contribute to its pathogenicity; these include haemagglutination and trypsin-like protease activity. Previous studies suggest an association between haemagglutination and trypsin-like protease activity of P. gingivalis. To investigate this, two complementary quantitative experimental approaches were taken. Five independent mutants of P. gingivalis deficient in trypsin-like protease activity were shown to exhibit reduced haemagglutination activity. In addition, enhancers (cysteine and dithiothreitol) and inhibitors (N-ethylmaleimide, N-p-tosyl-L-lysine-chloromethyl ketone, and phenylmethylsulphonyl fluoride) of trypsin-like protease activity were shown, respectively, to significantly enhance and inhibit haemagglutination activity of washed, wild-type P. gingivalis cells (p less than 0.05, paired t-test). Statistical analysis indicated a strong correlation between haemagglutination and trypsin-like protease activity (r = 0.85, p less than 0.001, Spearman rank correlation). The effect of the protease enhancers and inhibitors on haemagglutination activity was specific for P. gingivalis, as they did not significantly change the haemagglutination activity of Fusobacterium nucleatum. These results suggest that the proteolytic site of the trypsin-like protease participates in haemagglutination activity of P. gingivalis.

Animals↗

PubMed1977

Radioimmunoassay procedure for quantitating bacterial antibody in human sera.

A RIA system was developed to detect antibodies in human sera against bacteria. Sonicates of Escherichia coli and Fusobacterium polymorphum were used as antigens to sensitize plastic-coated beads; antibodies to these antigens were detected with 125I-labeled antihuman globulin. Serum antibody titers against E. coli were determined by the serial dilution method; from the results the standard curve principle was applied in determining the relative amounts of antibodies against E. coli in serum samples tested at a single dilution. The coefficient of variation of the RIA procedure was less than 10%. Serum titers obtained by the RIA and indirect immunofluorescence test were compared; RIA was more sensitive, quantitative and objective. Absorption studies, using E. coli and F. polymorphum absorbentes against E. coli and F. polymorphum. This RIA procedure offers a combination of desirable advantages; it is sensitive, specific, objective, quantitative, and easy to perform.

Antibodies, Bacterial↗

PubMed1979

Ultrastructural localisation of lipopolysaccharide-binding sites with peroxidase-conjugated lipopolysaccharides.

The localisation of lipopolysaccharide-binding sites on erythrocytes with peroxidase-coupled LPS is described. LPS was isolated from Fusobacterium nucleatum (Fus MC-8) by phenol-water extraction. The LPS was coupled to horseradish peroxidase by the two-step method of Avrameas and Ternynck (1971). The biological and serological activities of the conjugated LPS were compared with those of the native material. Peroxidase could be coupled to LPS without significant loss of endotoxic or serological activity. The LPS-peroxidase conjugate could be demonstrated on erythrocytes by light and electron microscopy.

Animals↗

PubMed1989

Quantitative bacteriology of a case of acute parotitis.

It is generally assumed that the microorganisms responsible for acute suppurative parotitis are facultative anaerobes, primarily Staphylococcus aureus and Streptococcus viridans. A quantitative microbiologic investigation of an aspirate of pus from the case we report revealed a microbial flora (5.1 X 10(6) CFU/ml) consisting entirely of strict anaerobes (Fusobacterium nucleatum and Peptostreptococcus anaerobius). The strain of F. nucleatum was sensitive to metronidazole but resistant to penicillin, amoxicillin, and erythromycin (minimum inhibitory concentration greater than 16 micrograms/ml). P. anaerobius was sensitive to the antibiotics tested. It is concluded that a high concentration of viable anaerobic bacteria may be present in acute suppurative parotitis and thus pus obtained from cases of sialadenitis should be cultured with microbiologic methods capable of isolating strict anaerobes. Additionally, antibiotic sensitivity should be determined routinely.

Acute Disease↗

PubMed1992

Effect of extraction of partly erupted third molars on subgingival microorganisms.

This study was made to investigate the effect of extraction of third molars on subgingival microbes in 39 generally and gingivally healthy men with an average age of 20.2 years (SD 0.9). Microbial samples were taken from the pericoronal space of symptom-free partly erupted lower third molars and from the adjacent gingival pockets of the second molars. The samples were cultivated anaerobically. All partly erupted third molars were extracted from 20 subjects. A control group of 19 subjects was left untreated. Microbe sampling was repeated 2 and 5 months postoperatively with highly significant results. It was shown that at baseline the number of black-pigmented gram-negative bacteria and Fusobacterium species was more frequent in third molar than in second molar sites. The total bacterial count decreased significantly at the second molar sites after extraction of the third molars when compared with the control group. Before the extractions, black-pigmented gram-negative bacteria were detected in 45% of the test subjects and Actinobacillus actinomycetemcomitans in 20%. The respective postoperative figures were 30% for black-pigmented gram-negative bacteria and 10% for Actinobacillus actinomycetemcomitans. Capnocytophaga species were not affected by the extractions. The findings suggest that erupting third molars may harbor harmful bacteria that can be reduced by eradicating the foci.

Adult↗

PubMed1992

Complement activation by lipopolysaccharides purified from gram-negative bacteria isolated from infected root canals.

The purpose of the present study was to investigate the complement activation by lipopolysaccharides purified from Porphyromonas (Bacteroides) endodontalis, Veillonella parvula, and Fusobacterium nucleatum isolated from infected root canals. The rate of consumption of the C3T component of the complement increased remarkably when lipopolysaccharides of more than 10 micrograms were added.

Bacteroides↗

PubMed1978

Azo reduction of trypan blue to a known carcinogen by a cell-free extract of a human intestinal anaerobe.

The azo reductase activity of a cell-free extract of Fusobacterium sp. 2 is characterized using trypan blue as a substrate. Either chemical reduction of this dye with sodium hydrosulfite or reduction by the cell-free extract produces a mutagenic product, o-tolidine. The o-tolidine is mutagenic in the Ames Salmonella/mammalian-microsome mutagenicity test when activated by a rat liver S9 preparation.

Benzidines↗