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Identification and antibiotic sensitivity of bacteria isolated from periapical lesions.

Periradicular tissues from 28 refractory endodontic cases requiring surgical intervention were submitted for histological diagnosis and microbiological culture. Bacteria isolated from these lesions were identified and then tested for their antibiotic sensitivity to a panel of common antibiotics. The periapical tissue specimens of 22 out of 28 lesions (79%) contained microorganisms. Of the 22 cases showing positive growth cultures, 15 were polymicrobial and 7 were single species isolates. Fifty-three different species were recovered: 29 anaerobes, 19 facultative anaerobes, and 5 aerobes. Microbes were observed under light microscopy in only one case. The most common organisms isolated were Propionibacterium acnes, Staphylococcus epidermidis, Streptococcus intermedius, Wolinella recta, Fusobacterium species, and Clostridium species. Antibiotic susceptibility results showed no clear cut evidence of significant antibiotic resistance among the species tested. The results of this study seem to corroborate earlier studies regarding the microbial population of periapical lesions refractory to nonsurgical endodontics.

Anti-Bacterial Agents↗

Anaerobic bacteria in dentoalveolar infections.

The bacteriology of 57 dentoalveolar infections was studied using optimal techniques to collect, transport and process specimens. There was an average of 4 bacterial species per specimen, and only 1/3 of the specimens held aerobes. Among the aerobic bacteria, streptococci dominated and among the anaerobes the Gram-negative rods, Bacteroides ruminicola and Fusobacterium nucleatum, were most frequently isolated followed by Gram-positive cocci, in particular Streptococcus intermedius. All aerobic isolates were resistant to penicillins but sensitive to clindamycin and tinidazole. The other anaerobic isolates were sensitive to penicillins but showed varying susceptibility to erythromycin and doxycycline. Tinidazole was effective against all anaerobic Gram-negative rods. The presence of volatile fatty acids in pus from dentoalveolar infections was found to be of presumptive value for the diagnosis of anaerobic infections. Direct gas-liquid chromatographic analysis of pus is recommended as a routine procedure for preliminary diagnosis of anaerobic dentoalveolar infections.

Adult↗

The micro-flora of the small bowel in health and disease.

The micro-flora of the proximal jejunum in healthy volunteers was compared with the micro-flora in patients with gastrointestinal symptoms suggestive of spontaneous bacterial overgrowth in the small intestine. Biopsies were taken distally to the ligament of Treitz with a Watson capsule. The samples were diluted and inoculated on selective and non-selective agar plates that were incubated aerobically and anaerobically. No major differences were found in the small jejunum micro-flora in healthy persons or in a heterogenous group of patients with gastrointestinal disorders. Oropharyngeal micro-organisms dominated the micro-flora in all subjects and colonic micro-organisms were found in low numbers in a few subjects from both groups. Streptococcus intermedius and Haemophilus parahaemolyticus were only found in the micro-flora of healthy subjects while Lactobacillus spp. was more frequently found in the samples from patients. Eight of 20 healthy subjects and five of 18 patients met the criterion of small intestinal overgrowth. Emerging evidence suggests that other factors are involved in the pathogenesis of the irritable bowel syndrome complex. There is a need for better understanding of the complicated interactions between the host and the endogenous micro-flora.

Journal Article↗

Two cases of lateral sinus thrombosis presenting with extracranial head and neck abscesses.

Two cases of lateral sinus thrombosis in fit adults with no previous otological history are presented. One case occurred in association with Bezold's abscess and followed mastoiditis which was masked by previous antibiotic treatment. The other case occurred in association with an occipitoparietal scalp abscess and there was no obvious preceding middle ear infection. The causative bacteria were Streptococcus milleri and a variant Streptococcus intermedius. Lateral sinus thrombosis is discussed.

Abscess↗

Humoral immunity host factors in subjects with failing or successful titanium dental implants.

BACKGROUND: Treatment with titanium dental implants is in general successful. However, an unknown number of implants do not integrate and are removed either by exfoliation or at the time of second stage surgery. It would be of importance to identify subjects at risk and predict early implant failure. METHODS: In a retrospective study serum IgG antibody titers and avidity in sera from 40 subjects who had experienced titanium dental implant treatments with non-osseo-integration as the outcome (NOTI) and in sera from 40 age and gender matched control subjects who had received successful titanium dental implants (SOTI) were studied. Serum IgG titers to whole cell Actinomyces viscosus, Bacteroides forsythus, Porphyromonas gingivalis, Staphylococcus aureus, and Streptococcus intermedius sonicated antigen preparations were studied by ELISA. RESULTS: Serum IgG antibody titers to S. aureus were significantly higher in subjects with SOTI than in NOTI (p<0.001) suggesting that higher titers indicate protection against implant failure as a result of S. aureus infection. Statistically significant higher serum IgG antibody avidity to P. gingivalis and B. forsythus were found in subjects with SOTI than in subjects with NOTI (p<0.01 and p<0.001, respectively). Statistical analysis failed to demonstrate antibody titer or avidity differences to the other pathogens studied. The likelihood that SOTI was associated with a high OD reading for S. aureus was 13.1:1 (p<0.001). Whether subjects were edentulous or not, or if they had lost teeth because of periodontitis or caries did not seem to matter. CONCLUSION: Serum IgG antibodies relative to B. forsythus, P. gingivalis and S. aureus may be associated with the outcome of implant procedures and explain why early implant failures occur.

Adult↗

Human CD59 is a receptor for the cholesterol-dependent cytolysin intermedilysin.

Cholesterol is believed to serve as the common receptor for the cholesterol-dependent cytolysins (CDCs). One member of this toxin family, Streptococcus intermedius intermedilysin (ILY), exhibits a narrow spectrum of cellular specificity that is seemingly inconsistent with this premise. We show here that ILY, via its domain 4 structure, binds to the glycosyl-phosphatidylinositol-linked membrane protein human CD59 (huCD59). CD59 is an inhibitor of the membrane attack complex of human complement. ILY specifically binds to huCD59 via residues that are the binding site for the C8alpha and C9 complement proteins. These studies provide a new model for the mechanism of cellular recognition by a CDC.

Animals↗

Infection-stimulated infraosseus inflammation and bone destruction is increased in P-/E-selectin knockout mice.

Infections of the dental pulp commonly result in infraosseus inflammation and bone destruction. However, the role of phagocytic leucocytes in the pathogenesis of pulpal infections has been uncertain. In this work we used P/E-/- selectin-deficient mice, which lack rolling adhesion of leucocytes to endothelium and mimic the human syndrome, leucocyte adhesion deficiency II (LAD-II), to test the hypothesis that phagocytic leucocytes protect against pulpal infection and subsequent periapical infraosseus bone resorption. P/E-/- mice and P/E+/+ wild-type controls were subjected to surgical pulp exposure, and both groups were infected with a mixture of pulpal pathogens including Prevotella intermedia, Fusobacterium nucleatum, Peptostreptococcus micros and Streptococcus intermedius. Animals were killed after 20 days, and the extent of infraosseus bone destruction was quantified by histomorphometry. In two separate experiments, P/E-/- mice had significantly greater bone resorption than P/E+/+ controls. The increased bone destruction correlated with a twofold decrease in polymorphonuclear (PMN) infiltration into periapical inflammatory tissues of P/E-/- mice. P/E-/- mice had higher tissue levels of the bone resorptive cytokine, interleukin (IL)-1alpha. Tissue levels of IL-2, IL-4, IL-10, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) were all higher in P/E-/- mice, but the increases were not statistically significant. Only IL-12 was higher in P/E+/+ mice, possibly reflecting a greater number of infiltrating monocytes in wild-type mice. These findings demonstrate that phagocytic leucocytes are protective in this model, and suggest that elevated expression of inflammatory cytokines is responsible for the observed bone destruction.

Animals↗

Dental implant infections.

Dental implants provide a restorative tool to support crowns, bridge abutments, and removable dentures. Osseointegrated implants are titanium posts that are surgically implanted in alveolar bone. A tight immobile bond (osseointegration) forms between bone and titanium, and prosthetic and restorative fixtures are attached to the implants. Titanium implants differ from natural teeth, which may make them more susceptible to mechanical stress. A small proportion of implants are not successful and may fail due to infection. The microbiota of implants is similar to that of teeth in similar clinical states. Implants that fail because of mechanical stress are colonized by species associated with healthy teeth. Infected implants are colonized by subgingival species, including Porphyromonas gingivalis, Bacteroides forsythus, Fusobacterium nucleatum, Campylobacter gracilis, Streptococcus intermedius, and Peptostreptococcus micros. Different patients may be colonized by different microbial complexes, indicating that optimal treatment should be directed to the specific infection.

Bacterial Infections↗

Agar medium for gas-liquid chromatography of anaerobes.

This study evaluates a method of performing gas-liquid chromatography (GLC) by direct extraction of fatty acids from agar for identification of clinically significant anaerobic bacteria. The potential use of agar cultures for GLC was studied by comparing chromatograms of 117 clinically isolated anaerobes grown in peptone yeast glucose broth and chopped meat carbohydrate broth, and on enriched brucella blood agar. For 98 of 117 anaerobes, fatty acid patterns from agar cultures were similar to those in broth. Significant differences were only found with Streptococcus intermedius, Clostridium perfringens, Clostridium tertium, and Actinomyces species, which produced less of certain fatty acids on agar than in broth. Results of this study indicate that GLC of short chain fatty acids produced on agar medium by anaerobes, combined with simple tests such as Gram's stain and colonial morphology, may allow fir direct presumptive genus identification from an initial pure agar culture.

Agar↗

The Streptococcus milleri group as a cause of pulmonary infections.

Streptococci that colonize the mouth and upper respiratory tract tend to be considered harmless commensals. In 45 cases of acute pneumonia and/or pulmonary abscess and 25 cases of thoracic empyema, the predominant species recovered were anaerobic bacteria and the Streptococcus milleri group, which encompasses the oral species Streptococcus anginosus, Streptococcus constellatus, and Streptococcus intermedius. The isolation of most S. milleri organisms along with oral anaerobes indicated synergy between these groups. Studies in a mouse model of pneumonia demonstrated this synergy; mortality was higher, histopathologic abnormalities were more marked (reflecting acute pneumonia followed by pulmonary abscess or empyema), and viable bacteria were more numerous in the lungs of mice with mixed infections caused by the S. milleri group and anaerobes than in the lungs of those with monomicrobial infection. In vitro studies elucidated a possible mechanism of this synergistic effect: anaerobes may enhance the growth of the S. milleri group and/or inhibit the bactericidal activity of the host. We conclude that the S. milleri group is more important in pulmonary infections than has previously been recognized.

Animals↗

Risk factors for nosocomial pneumonia in critically ill trauma patients.

OBJECTIVE: To determine risk factors for nosocomial pneumonia in critically ill trauma patients. DESIGN: Prospective cohort study. SETTING: The trauma intensive care unit (ICU) of a 1500-bed tertiary-care hospital. PATIENTS: All critically ill trauma patients (n = 103) admitted consecutively between November 1995 and October 1996. INTERVENTIONS: A comparison of data recorded at the time of ICU admission and during the clinical evolution in patients with (n = 23) and without (n = 80) nosocomial pneumonia was made. Data referred mainly to possible risk factors were recorded; they also included factors related to pneumonia etiology and evolutive factors. Predictors of nosocomial pneumonia were assessed by logistic regression analysis. MEASUREMENTS AND MAIN RESULTS: The presence of significant growth on quantitative cultures of the protected specimen brush (> or = 103 colony forming units/mL) was required to accept pneumonia as microbiologically proven, as well as the concurrence of a cohort of clinical and radiologic signs. Twenty-three (22.3%) patients developed nosocomial pneumonia. The mean age of these patients was 41.7 yrs; 18 of them (78.3%) were men. The microorganisms isolated in significant concentrations were Acinetobacter baumanii (ten cases), Staphylococcus aureus (11 cases), Pseudomonas aeruginosa (five cases), Haemophilus influenzae (two cases), and Klebsiella pneumoniae, Citrobacter freundii, Serratia marcescens, Enterococcus spp., Enterobacter spp., coagulase-negative Staphylococcus, and Streptococcus intermedius (one case each one). Risk factors for pneumonia by univariate analysis included nasogastric tube; continuous enteral feeding; prolonged mechanical ventilation (>1 day); use of H2-receptor antagonist, sucralfate, muscle relaxants, corticosteroids, barbiturates, and inotropic agents; positive end-expiratory pressure; intense sedation; re-intubation; tracheotomy; urgent brain computed tomography (CT) scan; craniotomy; iatrogenic event; and hyperventilation. The mortality rate was 43.5% (10 of 23) in the nosocomial pneumonia group and 18.8% in patients without nosocomial pneumonia (p =.02). Also, the mean stay in the ICU, the therapeutic charge (measured with total and mean punctuation of the Therapeutic Intervention Scoring System) and the complications, infectious and noninfectious, of the clinical evolution were significantly more frequent in patients with nosocomial pneumonia than in those without pneumonia (p <.05). In the multivariate analysis, continuous enteral feeding, craniotomy, prolonged mechanical ventilation (>24 hrs), use of positive end-expiratory pressure, and corticotherapy were independent predictors of nosocomial pneumonia. CONCLUSIONS: It seems that factors related to the patient's clinical course, rather than variables registered on the first days of ICU admission, are those that would exert an influence on the development of nosocomial pneumonia in critically ill trauma patients. In this way, from our point of view, in our study the main risk factors are the use of prolonged mechanical ventilation (>4 hrs) and positive end-expiratory pressure. At the same time, we can conclude that the reduction of this infection incidence could decrease the mean stay in the ICU, the therapeutic charge, and the prognosis in terms of mortality and morbidity.

Adult↗

Pott's puffy tumor and epidural abscess arising from dental sepsis: a case report.

OBJECTIVE: To present an unusual case of two uncommon cranial complications of frontal sinusitis: Pott's puffy tumor and epidural abscess arising from frontal sinusitis of dental origin, and also two systemic complications of sinusitis: septicemia and empyema, all occurring in an immunocompetent patient. STUDY DESIGN: A 21-year-old man presented with a scalp swelling and epidural abscess. Magnetic resonance imaging and computed tomographic scans revealed unilateral opacification of the frontal sinus and an epidural abscess with a direct connection to the scalp abscess. Further history revealed that his symptoms occurred coincidentally with a tooth extraction 2 months before, and he was hospitalized soon after the tooth extraction for sepsis and a lung abscess. METHODS: A combined neurosurgical and otolaryngologic approach was required to treat the sinusitis and the associated epidural and scalp abscess. RESULTS: Cultures returned as Streptococcus intermedius from all three sites. The patient was free of disease at the 3-month follow-up. CONCLUSIONS: Odontogenic maxillary sinusitis is well documented; however, there is little reported of frontal sinusitis arising from dental disease. The prevalence of sinusitis of dental origin will be reviewed, including the microbiology of this particularly virulent organism that persisted despite earlier treatment with ampicillin. Also, the current thoughts on management of these cases will be discussed with particular reference to local therapy for sinusitis in addition to systemic treatment with antibiotics.

Abscess↗

Haemophilus segnis polymicrobial and monomicrobial bacteraemia identified by 16S ribosomal RNA gene sequencing.

This paper reports a case of Haemophilus segnis polymicrobial bacteraemia and a case of H. segnis monomicrobial bacteraemia identified by 16S ribosomal RNA gene sequencing. In the first case, a gram-negative aerobic coccobacillus was isolated with Streptococcus intermedius and S. sanguis from the blood culture of a 32-year-old intravenous drug addict with left thoracic empyema. In the second case, a gram-negative aerobic coccobacillus was isolated from the blood culture of an 82-year-old woman with Clostridium difficile colitis and septicaemic shock. Both gram-negative coccobacilli grew on chocolate agar as colonies of 1 mm in diameter after incubation for 24 h at 37 degress C in air with CO2 5%, but only to pinpoint sizes on blood agar under the same incubation conditions. Both strains were factor V-dependent, but not factor X-dependent. For the first isolate, the Vitek system (NHI) showed that it was 56% likely to be Actinobacillus actinomycetemcomitans and 40% Neisseria subflava; whereas the API system (NH) showed that it was 58% likely to be H. aphrophilus/paraphrophilus and 42% H. parainfluenzae. For the second isolate, the Vitek system (NHI) showed that it was 95% likely to be H. influenzae VIII; whereas the API system (NH) showed that it was 58% likely to be H. aphrophilus/paraphrophilus and 42% H. parainfluenzae. 16S rRNA gene sequencing showed that there were four base differences between isolate 1 and H. segnis and two base differences between isolate 2 and H. segnis, indicating that both isolates most closely resembled a strain of H. segnis. Only two cases of H. segnis bacteraemia were found in the English scientific literature, one in a case of infective endocarditis and the other in a case of pancreatic abscess. Including the present two cases, the overall mortality of H. segnis bacteraemia was 50%.

Adult↗

Peripheral lymphoid hyperplasia and central lymphoid depletion in mice treated with a bacterial B-cell mitogen (F3'EP-Si/p90).

In order to further understand the mechanism mediating the mitogenic and immunosuppressor effects of p90, a protein produced by Streptococcus intermedius, flow cytometric studies were performed on peripheral and central lymphoid organs of mice treated with this protein. p90 induced a strong blastogenic B-cell response in the spleen and lymph nodes, followed by a slight but significant polyclonal T-cell activation. B-cell repertoire analysis indicated that polyclonal B-cell responses affected similarly both CD5+ and conventional (CD5-) B cells in the spleen. Repertoire analysis of T cells failed to reveal any preferential stimulation of the V beta T-cell receptor (V beta-TcR) families studied. Peripheral lymphoid hyperplasia was observed concomitantly with central lymphoid depletion. In the bone marrow, pre-B and B cells were profoundly depleted, with a more pronounced effect on small pre-B cells. In the thymus, double-positive (CD4+CD8+) thymocytes were preferentially eliminated, with a relative enrichment of single positive (either CD4+ or CD8+) and double-negative (CD4-CD8-) thymocytes.

Animals↗

Effect of khat chewing on 14 selected periodontal bacteria in sub- and supragingival plaque of a young male population.

BACKGROUND/AIMS: The habit of chewing khat (Catha edulis) for its amphetamine-like effects is highly prevalent in Yemen and east Africa, and has expanded to Western countries. The purpose of this study was to estimate and compare the prevalence and levels of 14 periodontal bacteria in gingival plaque of khat chewers and khat nonchewers, as well as of khat chewing sides and khat nonchewing sides. METHODS: A total of 408 sub- and supragingival plaque samples were collected from 51 young males (29 khat chewers and 22 khat nonchewers; age range 19-28 years) and analyzed using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Clinical parameters were recorded for all teeth at six sites per tooth. RESULTS: Streptococcus intermedius and Veillonella parvula were significantly more prevalent in the subgingival plaque of chewers, which also showed significantly higher levels of V. parvula and Eikenella corrodens. Similar results were found for the subgingival plaque of the chewing sides compared to the nonchewing sides. However, there was a significantly higher prevalence and higher levels of Tannerella forsythia in the subgingival plaque of the nonchewing sides. No significant differences were observed for the supragingival plaque between the two study groups. There was a significantly lower prevalence of Capnocytophaga gingivalis and Fusobacterium nucleatum in the khat chewing sides, and higher levels of V. parvula and Actinomyces israelii. CONCLUSION: The data suggest that khat chewing induces a microbial profile that is not incompatible with gingival health.

Adult↗

Association of Eubacterium nodatum and Treponema denticola with human periodontitis lesions.

BACKGROUND: The purpose of the present investigation was to compare the levels, proportions and percentage of sites colonized by 40 bacterial species in subgingival plaque samples from periodontally healthy subjects and patients with chronic periodontitis to seek possible pathogens other than the consensus pathogens Porphyromonas gingivalis and Tannerella forsythia. METHOD: Subgingival plaque samples were taken from the mesial aspect of each tooth in 635 subjects with chronic periodontitis and 189 periodontally healthy subjects. The samples were individually analyzed for their content of 40 bacterial species using checkerboard DNA-DNA hybridization (total samples = 21,832). Mean counts, % DNA probe counts and percentage of sites colonized at >10(5) were determined for each species in each subject and then averaged in each clinical group. Significance of difference between groups was determined using the Mann-Whitney test. Association between combinations of species and periodontal status was examined by stepwise logistic regression analysis. Analyses were repeated using a subset of subjects from both clinical groups who had proportions of P. gingivalis plus T. forsythia less than the median (4.42%) found in periodontally healthy subjects. All analyses were adjusted for multiple comparisons. RESULTS: For the 824 subjects the consensus pathogens P. gingivalis and T. forsythia as well as Eubacterium nodatum and Treponema denticola had significantly higher mean counts, proportions and percentage of sites colonized in samples from subjects with periodontitis than from periodontally healthy subjects. There were significantly more Capnocytophaga gingivalis, Streptococcus gordonii and Veillonella parvula in periodontally healthy subjects. E. nodatum, T. denticola, Streptococcus oralis, Streptococcus intermedius, Fusobacterium nucleatum ssp. vincentii all had higher counts and proportions in diseased than healthy subjects who had low proportions of P. gingivalis and T. forsythia. Logistic regression analysis indicated that the same species groups were associated with disease status after adjusting for the proportions of the other species. CONCLUSIONS: This investigation confirmed the strong association of P. gingivalis and T. forsythia with chronic periodontitis and emphasized a strong association of E. nodatum and T. denticola with periodontitis whether in the presence or absence of high levels of the consensus pathogens. Other species, including S. oralis, Eikenella corrodens, S. intermedius and F. nucleatum ssp. vincentii, were associated with disease when P. gingivalis and T. forsythia were present in low proportions.

Adult↗

Frequency distributions of periodontal attachment loss. Clinical and microbiological features.

The present investigation attempted to determine if the pattern of past periodontal destruction could be concisely summarized, and related to other clinical and microbiological parameters. 61 subjects between the ages of 12 and 61 years with destructive periodontal disease were evaluated at 6 sites per tooth for redness, plaque, suppuration, bleeding on probing, pocket depth, and attachment level. The frequency distribution of baseline attachment level measurements was computed for each individual. A curve fitting algorithm was used to fit the frequency distribution to 1-, 2-, and 3-term normal distributions. The parameters of the fit could be used to summarize concisely all of the frequency distributions. 3 major patterns of attachment loss could be distinguished. Pattern I required a two-term distribution with localized destruction at less than 34% of sites and was further divided into 3 groups, depending on average attachment loss at diseased sites. The means of the second peak for the subgroups were 2.7, 5.3, and 8.6 mm, respectively. Pattern II exhibited more widespread disease (greater than 33% of sites affected) with multiple peaks in the frequency distribution requiring a 3-term distribution for satisfactory fit. However, a significant proportion of sites was not affected. Pattern III exhibited a single-peaked normal distribution in which virtually all sites were affected. Mean attachment levels of the peaks in this group ranged from 2.7 to 8.4 mm. 23 of the 61 subjects showed significant attachment loss at 1 or more sites during the course of bi-monthly monitoring, as determined by the tolerance method of analysis. Subgingival plaque samples were taken from these active sites and matched with control sites prior to therapy. The proportions of Fusobacterium nucleatum, Streptococcus intermedius, and Eikenella corrodens were significantly elevated in active and control sites of subjects in groups II and III combined (the widespread disease groups), and proportions of Actinobacillus actinomycetemcomitans and Propionibacterium acnes were elevated in active and control sites of the more localized disease group I subjects. Group I subjects showed a 13- to 15-fold decrease in hazard rates of periodontal sites after Widman flap surgery and systemic tetracycline, whereas groups II and III subjects showed 2-to 6-fold decreases.

Adolescent↗

Multiple periodontal abscesses after systemic antibiotic therapy.

Multiple periodontal abscesses were reported in medically compromised patients. We examined patients with a non-contributory medical history referred for the treatment of numerous periodontal abscesses. All patients had taken oral broad spectrum antibiotics 1 to 3 weeks prior to the outburst of the abscesses (8 patients: penicillin, 2 patients: tetracycline). The patients suffered from advanced periodontal disease, 82% of the examined sites showed probing depths greater than 3 mm, 56% attachment loss greater than 3 mm. Subgingival plaque samples were analysed from 2 different abscess sites. Bacteroides gingivalis (19/20), Fusobacterium nucleatum (13/20) and Streptococcus intermedius (13/20) were the most prevalent anaerobic microbiota. Strains resistant to the prescribed antibiotic were found in 55% (11/20) of the subgingival plaque samples. It was concluded that in patients with advanced periodontal disease, systemic antibiotic therapy without subgingival debridement may change the composition of the subgingival microbiota, thus favouring the outburst of multiple periodontal abscesses.

Adult↗