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Occurrence of invading bacteria in radicular dentin of periodontally diseased teeth: microbiological findings.

Bacterial invasion in roots of periodontally diseased teeth, which has been recently documented using cultural and microscopic techniques, may be important in the pathogenesis of periodontal disease. The purpose of this investigation was to determine the occurrence and the species of invading bacteria in radicular dentin of periodontally diseased teeth. Samples were taken from the middle layer of radicular dentin of 26 periodontally diseased teeth. 14 healthy teeth were used as controls. Dentin samples were cultured anaerobically. The chosen methodology allowed the determination of the numbers of bacteria present in both deeper and outer part of dentinal tubules, and the bacterial concentration in dentin samples, expressed as colony forming units per mg of tissue (CFU/mg). Invading bacteria was detected in 14 (53.8%) samples from periodontally diseased teeth. The bacterial concentration ranged from 831.84 to 11971.3 CFU/mg (mean+/-standard deviation: 3043.15+/-2763.13). Micro-organisms identified included putative periodontal pathogens such as Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium nucleatum, Bacteroides forsythus, Peptostreptococcus micros and Streptococcus intermedius. These findings suggest that radicular dentin could act as bacterial reservoir from which periodontal pathogens can recolonize treated periodontal pockets, contributing to the failure of therapy and recurrence of disease.

Adult↗

Protein degradation by Prevotella intermedia and Actinomyces meyeri supports the growth of non-protein-cleaving oral bacteria in serum.

The proteolytic activities of oral bacteria are thought to play an important role in the aetiology of dental abscesses. Bacteria-derived proteases may contribute to tissue destruction, and are likely to impair host defence by degrading immunoglobulins and complement. Degraded periodontal tissue and tissue fluid are likely to constitute essential sources of nutrients in the abscess. Tissue fluid, which is derived from serum, is rich in protein and poor in carbohydrate, suggesting that breakdown of protein and fermentation of amino acids is a crucial step to generate energy for growth of the microflora. The number of oral bacterial species that perform hydrolytic cleavage of protein into polypeptides, the first step in protein degradation, is relatively small compared to the large majority of peptidase-producing species. In this study, we therefore investigated the growth-promoting effect of proteinase-producing species like Prevotella intermedia and Actinomyces meyeri on the growth of some non-proteinase producing bacteria in mixed cultures. We used serum as a substitute for the supposed natural substrate of the abscess microflora. The breakdown of serum proteins was investigated using capillary electrophoresis. Poor growth was found in mono- and mixed cultures of non-proteinase producing species Eubacterium lentum, Fusobacterium nucleatum. Peptostreptococcus micros, and Streptococcus intermedius. The presence of P. intermedia in mixed cultures strongly enhanced growth of these 4 species, according to the hypothesis that the growth of the mixed cultures was peptide-limited. The enhanced growth of P. intermedia in pronase-digested serum indicated peptide-limited growth of this organism in serum, despite its production of proteinase. We found that growth of monocultures of Actinomyces meyeri was poor. In contrast, A. meyeri grew well in mixed cultures and its presence stimulated growth of F. nucleatum and P. micros, suggesting a synergistic relationship. The growth of mono- and mixed cultures was investigated using one representative strain of each species. Thus, there is a small risk of having selected unique strains. Proteinase inhibitors reduced the growth of Porphyromonas gingivalis, Prevotella nigrescens, and P. intermedia in trypticase peptone-yeast extract medium with, and without, IgG. Our study indicated that proteinase-producing organisms play a key role in mixed cultures of oral bacteria in human serum by providing polypeptides for growth. This may explain their association with dental abscesses.

Actinomyces↗

Periodontal infections and pre-term low birth weight: a case-control study.

OBJECTIVE: Pre-term delivery of low-birth-weight infants [pre-term low birth weight (PLBW)] remains a significant public health issue and a major cause of neonatal death and long-term health problems. There is a growing consensus that infections remote from fetal-placental unit may influence PLBW infants. Recent studies have suggested that maternal periodontal disease may be an independent risk factor for PLBW. The purpose of the present study was to evaluate the possible link between periodontal infections and PLBW by means of clinical and microbiological data in post-partum women with low socioeconomic level. METHODS: Clinical periodontal recordings comprising dental plaque, bleeding on probing, probing pocket depth and gingival recession were performed (six sites/tooth) in a total number of 181 women (53 cases and 128 controls) within 3 days post-partum. Subgingival plaque samples from mesio-or disto-buccal aspect of randomly selected one first molar and one incisor tooth have been obtained by paperpoints and were analysed by checkerboard DNA-DNA hybridization with respect to 12 bacterial species. In all analyses, the individual subject was the computational unit. Thus, mean values for all clinical parameters were calculated and bacterial scores from each individual sample were averaged. Statistical methods included Student's t-test, Fisher's exact test/chi(2) test, and multiple logistic regression analysis. RESULTS: The cases have gained significantly less weight during the pregnancy than did the controls (p<0.05). There were no statistically significant differences between the cases and controls with regard to the dental and periodontal parameters and the values of clinical periodontal recordings were found to be very similar (p>0.05). Mean and median scores (bacterial loads) of Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Actinobacillus actinomycetemcomitans, and Streptococcus intermedius in the subgingival plaque sampling sites were significantly higher in the controls than in the cases (p<0.05). The occurrence rates of P. intermedia, Fusobacterium nucleatum, Peptostreptococcus micros, Campylobacter rectus, Eikenella corrodens, Selenomonas noxia and S. intermedius were higher in the cases compared with the controls, but the differences were not statistically significant (p>0.05). According to the model created by the multiple logistic regression analysis, P. micros and C. rectus were found to significantly increase the risk of PLBW (p<0.01 and p<0.05 respectively), while P. nigrescens and A. actinomycetemcomitans decreased this risk (p<0.01). CONCLUSION: The present findings indicated that when subgingival bacteria were evaluated together, P. micros and C. rectus may have a role in increasing the risk for PLBW, although no single bacteria exhibited any relation with the risk of PLBW. Further studies are required to better clarify the possible relationship between periodontal diseases and PLBW.

Adolescent↗

Tobacco smoking and periodontal microflora in a Saudi Arabian population.

OBJECTIVES: To explore the subgingival periodontal microflora in a Saudi Arabian population with a special focus on its relationship with various smoking habits. MATERIAL AND METHODS: A total of 198 individuals in the age range 17-60 years were included in the study. 29% were water-pipe smokers, 18% cigarette smokers, 13% smokers of both water pipe and cigarettes (mixed smokers) and 40% non-smokers. For each individual, a subgingival plaque sample from the deepest site in each quadrant was obtained. The checkerboard DNA-DNA hybridization technology was used to determine the presence of Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Tannerella forsythensis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Treponoma denticola, Peptostreptococcus micros, Campylobacter rectus, Eikenella corrodens, Selenomonas noxia and Streptococcus intermedius. Two cut-off levels for detection were used, score 1(10(5) bacteria) and score 3 (10(6) bacteria). RESULTS: The prevalence of individuals positive for the different microorganisms at score 1 cut-off varied from 7% to 95%. At score 3 cut-off the prevalence varied from 0% to 30%. The depth of sample site was a key factor for detection. When the depth of sample site was taken into account, no statistically significant differences were observed between cigarette smokers, water-pipe smokers, and non-smokers with regard to occurrence of the microorganisms studied. CONCLUSIONS: No major differences were observed between cigarette smokers, water-pipe smokers, and non-smokers regarding the occurrence of the periodontal microorganisms studied suggesting that this portion of the subgingival periodontal microflora is independent of tobacco smoking.

Adolescent↗

[Clinical evaluation of lung abscess diagnosed by transtracheal aspiration].

We have diagnosed lung abscess according to findings of infiltration with cavity formation on chest X-ray and/or CT-scan and pathogens isolated from transtracheal aspirates. We evaluated the clinical features of 20 patients with lung abscess (18 males and 2 females, mean age; 54.3 years). Diabetus mellitus and periodontal diseases were prominent underlying diseases in patients with lung abscess. Cough was complained in 13 patients, chest or back pain in 9, purulent sputum in 8 and hemosputum in 5 when the patients admitted to our hospital. A temperature higher than 38 degrees C was present in 12 patients but temperature les than 37 degrees C in 2. Multiple microorganisms were cultured from TTA in 15 patients. A mean of 2.7 bacterial species per patient was isolated, aerobes alone being isolated in 2 patients, anaerobes alone in 3, and mixed aerobic and anaerobic isolates in 10. Seventeen strains of aerobes and 35 of anaerobes were isolated. Major pathogens were Streptococcus pneumoniae, Streptococcus intermedius and other in aerobes, and Peptostreptococcus micros, Fusobacterium necrophorum, Prevotella melaninogenica and others in anaerobes. Abnormality of chest X-ray was located on the right upper lobe in 6 patients, the right lower lobe in 6, the left upper lobe in 6, the left lower lobe in 4 and the right middle lobe in 1. All patients were cured only by treatment of antimicrobial agents, but cavity formation on chest X-ray remained in 4 patients after the treatment.

Adult↗

Experimental otitis media in gerbils and chinchillas with Streptococcus pneumoniae, Haemophilus influenzae, and other aerobic and anaerobic bacteria.

To ascertain the usefulness of Mongolian gerbils as an inbred model for otitis media, 52 Mongolian gerbils (Meriones unguiculatus, strain MONT/Tum) were compared with 26 chinchillas (Chinchilla laniger) for susceptibility to Streptococcus pneumoniae type 3. Haemophilus influenzae type b, and a polymicrobic culture including anaerobes (Streptococcus intermedius, Propionibacterium acnes, Staphylococcus epidermidis, and Corynebacterium sp.). Organisms were inoculated percutaneously into the superior chamber of the middle ear bulla. The gerbils and chinchillas shared similar susceptibilities and responses to the inoculated organisms as determined by X-ray, otoscopic, histopathological, and microbiological determinations at 5 to 7 days. Koch's postulate studies proved the role of S. pneumoniae and H. influenzae in the pathology found in both animal models. The animals were also susceptible to the polymicrobic culture, although the relative virulence of the individual members of this mixture was low, suggesting that these species potentiated as a polymicrobic mixture. The Corynebacterium sp. appeared to elicit the greatest histopathological response in chronic (8-week) studies in gerbils. The gerbils were found to be useful as an alternative animal model for the study of otitis media of bacterial etiology.

Animals↗

Natural history of aortic valve endocarditis in rats.

Sterile aortic vegetations were produced in rats by introducing a polyethylene catheter through the right carotid artery. The catheter was either left in place throughout the experiments or removed before bacterial challenge. Bacterial endocarditis was uniformly produced by intravenous injection of 10(7) colony-forming units of Staphylococcus aureus or Streptococcus intermedius, whether the catheter was left in place or removed. However, in rats with the catheter left in place, bacterial multiplication within the vegetations with both strains was accelerated, and mortality from Staphylococcus aureus infection was increased. Using 10(7) colony-forming units of serum-resistant Escherichia coli as a test microorganism, we found a marked difference in the production of endocarditis depending upon whether the catheter was left in place or removed before injection; only those animals infected with the catheter in place developed infection. From these experiments in rats, it was evident that the presence of a foreign body has a considerable influence on the ability of bacteria to grow within an intravascular vegetation. In addition, a striking difference in the virulence of the three strains studied was established; Staphylococcus aureus was the most, and E. coli the least, pathogenic.

Animals↗

B-Cell deficiency predisposes mice to disseminating anaerobic infections: protection by passive antibody transfer.

We have previously demonstrated that a high proportion of RAG-2 SCID knockout mice, which lack T and B cells, develop orofacial abscesses and disseminated infections following pulpal infection, whereas immunocompetent control mice do not. In the present study, we sought to identify the components of the adaptive immune response which contribute to protection against disseminating anaerobic infections and sepsis. For this purpose, various genetically engineered immunodeficient mice were employed, including RAG-2 SCID, Igh-6 (B-cell deficient), Tcrb Tcrd (T-cell deficient) and Hc(0) (C5 deficient). For abscess induction, the mandibular first molars were subjected to pulp exposure on day 0. Teeth were infected with a mixture of four anaerobic pathogens, including Prevotella intermedia, Streptococcus intermedius, Fusobacterium nucleatum, and Peptostreptococcus micros, and teeth were sealed to prevent communication with the oral cavity. The findings demonstrate that both RAG-2 SCID and B-cell-deficient mice, but not T-cell- or C5-deficient mice, have increased susceptibility to the development of disseminating anaerobic infections. Abscess-susceptible RAG-2 SCID and B-cell-deficient mice also showed a significant loss of body weight, splenomegaly, and absent antibacterial antibody production. Furthermore, dissemination was significantly reduced, from 74 to 25%, in susceptible RAG-2 mice by passively transferred antibody, predominantly immunoglobulin G2b (IgG2b) and IgM, against the infecting bacterial innoculum. Fractionated IgG-enriched preparations were more efficient in transferring protection than IgM preparations. We conclude that an antibody-mediated mechanism(s), most likely bacterial opsonization, is of importance in localizing anaerobic root canal infections and in preventing their systemic spread.

Abscess↗

Interleukin-6 deficiency increases inflammatory bone destruction.

Periapical bone destruction occurs as a consequence of pulpal infection. In previous studies, we showed that interleukin-1 (IL-1) is the primary stimulator of bone destruction in this model. IL-6 is a pleiotropic cytokine that is induced in these infections and has both pro- and anti-inflammatory activities. In the present study, we determined the role of IL-6 in regulating IL-1 expression and bone resorption. The first molars of IL-6 knockouts (IL-6(-/-)) and wild-type mice were subjected to surgical pulp exposure and infection with a mixture of four common pulpal pathogens, including Prevotella intermedia, Fusobacterium nucleatum, Peptostreptococcus micros, and Streptococcus intermedius. Mice were killed after 21 days, and bone destruction and cytokine expression were determined. Surprisingly, bone destruction was significantly increased in IL-6(-/-) mice versus that in wild-type mice (by 30%; P < 0.001). In a second experiment, the effects of chronic (IL-6(-/-)) IL-6 deficiency and short-term IL-6 deficiency induced by in vivo antibody neutralization were determined. Both IL-6(-/-) (30%; P < 0.001) and anti-IL-6 antibody-treated mice (40%; P < 0.05) exhibited increased periapical bone resorption, compared to wild-type controls. The increased bone resorption in IL-6-deficient animals correlated with increases in osteoclast numbers, as well as with elevated expression of bone-resorptive cytokines IL-1alpha and IL-1beta, in periapical lesions and with decreased expression of the anti-inflammatory cytokine IL-10. These data demonstrate that endogenous IL-6 expression has significant anti-inflammatory effects in modulating infection-stimulated bone destruction in vivo.

Animals↗

Antigenic relationships and rapid identification of Peptostreptococcus species.

Antisera against whole cells of each Peptostreptococcus species (P. anaerobius, P. micros, P. parvulus, and P. productus) were produced in rabbits. When these antisera were reacted against sonically disrupted cells and culture supernatant fluids in Ouchterlony tests, lines of identity were obtained among the antigens from all the species and uninoculated culture medium. When the antisera were subsequently absorbed with the dehydrated culture medium used to grow the peptostreptococci, all cross-reactions in heterologous antigen-antibody combinations were eliminated, leaving only species-specific precipitin arcs. These absorbed antisera, specific for each Peptostreptococcus species by Ouchterlony tests, were used for rapid identification studies. Staphylococcus aureus-bearing protein A was sensitized with each absorbed antiserum. These reagents produced specific coagglutination reactions with suspensions of each Peptostreptococcus reference strain and with 16 clinical isolates. No cross-reactions occurred with the Streptococcus intermedius, Peptococcus magnus, or Peptococcus asaccharolyticus strains tested.

Agglutination Tests↗

Comparison of three methods for anaerobe identification.

In this study we evaluated the ability of three commercial methods, API 20A (Analytab Products, Plainview, N.Y.), Minitek (BBL Microbiology Systems, Cockeysville, Md.), and Anaerobe-Tek (Flow Laboratories, Inc., McLean, Va.), to accurately identify 165 recent clinical and 38 stock isolates of anaerobic bacteria without supplemental tests or gas-liquid chromatography. Strains included 89 Bacteroides spp., 12 fusobacteria, 10 gram-positive, nonsporing rods, 43 Clostridium spp., 15 Streptococcus intermedius, 18 peptococci, 6 peptostreptococci, 3 Staphylococcus saccharolyticus, and 7 Veillonella spp. The methods used were those of manufacturers, without supplemental tests. API 20A correctly identified 70.0% of strains to species and 6.4% to genus only, with 17.2% as part of a spectrum of identifications and 6.4% incorrect. Minitek, according to the current code book, yielded 69.5% correct identifications to species, 16.8% spectrum identifications, and 13.8% incorrect. Anaerobe-Tek correctly identified 64.0% of strains to species, 21.2% spectrum identifications, and 14.8% incorrect. Thirteen strains were misidentified by API 20A, 28 were misidentified by Minitek, and 30 were misidentified by Anaerobe-Tek. For laboratories without gas-liquid chromatography support and where identification of clinically significant Bacteroides fragilis and Clostridium perfringens is desired, any of the three systems would provide accurate information. For more extensive anaerobe identification, including the less frequently isolated, more unusual organisms, API 20A and Minitek are preferred at this time. All systems have identification schemes associated with a percentage of misidentifications, the most recently introduced Anaerobe-Tek system being associated with the highest error rate.

Bacteria, Anaerobic↗

Culture-independent identification of pathogenic bacteria and polymicrobial infections in the genitourinary tract of renal transplant recipients.

Renal transplant recipients are predisposed to urinary tract infections caused by both common uropathogens and opportunistic bacteria resulting frequently in significant polymicrobial infections. In this study, a culture-independent 16S rRNA-based approach was established to identify unusual, fastidious, or anaerobic bacteria and to investigate bacterial diversity in urinary tract specimens. Similarly sized amplicons encompassing the V6 to V8 region of the 16S rRNA were analyzed with denaturing high-performance liquid chromatography (DHPLC) (WAVE System). Artificial mixtures of single amplicons from commonly encountered uropathogenic bacteria produced distinct peak profiles whose identities were confirmed by sequencing individually collected peak products. We evaluated the application of the method on 109 urinary tract specimens from renal transplant recipients; 100% correlation was found for culture-positive specimens, and DHPLC generated peak profiles. However, for culture-negative specimens, DHPLC facilitated the detection of novel peak profiles. DNA sequencing of these individual peaks was used to identify the bacteria involved. Thus, in PCR-positive but culture-negative samples the method allowed detection of previously known uropathogens such as Corynebacterium urealyticum and Gardnerella vaginalis, but also unusual agents including Anaerococcus lactolyticus, Bacteroides vulgatus, Dialister invisus, Fusobacterium nucleatum, Lactobacillus iners, Leptotrichia amnionii, Prevotella buccalis, Prevotella ruminicola, Rahnella aquatilis, and Streptococcus intermedius were detected as single pathogens or as constituents of polymicrobial infections. The method described is reproducible and rapidly and enables both DHPLC-based profiling and sequence-based investigation of microbial communities and polymicrobial infections. A detailed understanding of infections found in recipients of renal transplants will guide antibiotic therapy regimens and provide new perspectives for decreasing the risk of graft rejection.

Bacteria↗

Survival of anaerobic bacteria in common laboratory diluents.

The survival of six species of anaerobic bacteria was studied in simple or commercially available diluents. Bacteroides fragilis and Fusobacterium nucleatum showed excellent survival in all diluents including distilled water. Fusobacterium mortiferum survived well in all diluents except water and water supplemented with 0.1% gelatain. Clostridium perfringens survived best in phosphate-buffered saline with gelatin. Peptococcus asaccharolyticus required gelatin added to the basic diluent, and Streptococcus intermedius showed excellent survival only in minimal essential medium with gelatin. These diluents could provide effective and economical alternatives to more complex and costly diluents often used in work with anaerobic bacteria.

Anaerobiosis↗

The Splendore-Hoeppli phenomenon in hepatic botryomycosis.

A 68 year old diabetic man developed septicaemia and multiple liver abscesses due to Streptococcus intermedius. Liver biopsy revealed streptococcal botryomycosis which showed the Splendore-Hoeppli phenomenon. Diabetes mellitus is an example of the immunosuppressed states that have been associated with botryomycosis.

Aged↗

[Psoas abscesses. An up-dated perspective].

OBJECTIVE: To analyze the characteristics of abscesses of the psoas muscle and assess the differences between pyogenic and tuberculous abscesses. METHODS: Retrospective descriptive study of all patients with psoas abscess in our hospital over the period 1994 to 2004. RESULTS: Fourteen patients were studied (10 males), with a mean age of 42 years. Half of them had had an underlying disease. The most frequent clinical manifestations were abdominal pain (64%), fever (57%), and back pain (43%). All the abscesses were secondary. In 7 patients the origin was gastrointestinal, in 6 osteoarticular, and 1 was related with infection of an aortobifemoral bypass. Computed tomography was the diagnostic imaging method in all patients. Culture of drainage specimens was positive in 92% of patients undergoing this procedure. Causal microorganisms included Mycobacterium tuberculosis (5), Streptococcus intermedius (4), Staphylococcus aureus (3), Escherichia coli (3) and Bacteroides fragilis (2). Tuberculous abscesses originated in spondylitis and the clinical presentation was longer prior to diagnosis. Drainage was performed in 12 patients (8 percutaneous and 4 surgical). Mean duration of antimicrobial therapy was 4 weeks. The infection resolved in all patients. CONCLUSIONS: Psoas abscess commonly had a gastrointestinal and osteoarticular origin. We underscore the high percentage of tuberculous etiology, which had a more insidious clinical and analytical presentation and was usually secondary to spondylitis. Prolonged antimicrobial treatment associated with drainage was effective therapy.

Adolescent↗

Recovery of anaerobic bacteria from a post-traumatic nasal septal abscess. A report of two cases.

This report presents 2 children (boys, 11 and 9 years old) who had post-traumatic nasal septal abscesses. Cultures for aerobic and anaerobic bacteria revealed Peptostreptococcuus magnus and a beta-lactamase-producing Prevotella intermedia in the first patient, and Peptostreptococcus anaerobius, Streptococcus intermedius, and Prevotella melaninogenica in the second patient. Anaerobic bacteria may play a role in nasal septal abscesses.

Abscess↗

Septic pulmonary embolism due to periodontal disease.

Three weeks following a toothache, a 56-year-old man developed cough, sputum, fever, and pleuritic chest pain. He had mild periodontal disease and his chest radiographs and chest computed tomographic (CT) scans showed multiple pulmonary nodules. The CT scan strongly suggested septic pulmonary embolism. Aspirated pus from one of the nodules yielded pure growth of Streptococcus intermedius. Lesions resolved with antimicrobial therapy. The usual predisposing factors for septic pulmonary embolism were absent, and, the isolation of S intermedius from the pus, the antecedent toothache, and periodontal disease all suggested the gingiva as the source. We hypothesize that periodontal infection led to bacteremia, seeding of the lungs, and multiple anaerobic pulmonary abscesses, akin to reported instances of infective endocarditis from dental foci without any prior dental procedures. To our knowledge, this presentation of septic pulmonary embolism is unprecedented.

Humans↗

Recolonization of the subgingival microflora after scaling and root planing in human periodontitis.

The purpose of this study was to evaluate the recolonization patterns of the subgingival microflora of adult periodontitis patients after a single session of scaling and root planing. In each of eight patients, three clinically diseased sites were investigated microbiologically by darkfield microscopy and cultural analysis. After initial clinical and microbiological parameters were determined, each subject received a single session of scaling and root planing but no oral hygiene instructions. Clinical indices were measured and microbial parameters were reassessed 7, 21, and 60 days after treatment in a manner such that each of the test sites was sampled only once after treatment. Recolonization was evaluated by matching any single site with its own preoperative site. A significant improvement in probing depth was noted for up to 60 days after treatment, while the gingival index did not change markedly during the course of the study. The microbial composition of treated sites 7 days after scaling and root planing, as determined by both cultural and darkfield data, was similar to that of periodontally healthy sites. Differences between cultural and darkfield data became apparent at the 21 day sampling point. The darkfield data showed that the sites consisted of cocci with few spirochetes. Cultural data demonstrated that the majority of the cocci were anaerobic, namely Streptococcus intermedius, Veillonella parvula, and Peptostreptococcus micros. At 60 days, there was no significant variation in any of the parameters from pretreatment levels. The most prevalent anaerobic rods prior to and 60 days after therapy were Fusobacterium nucleatum, Bacteroides gingivalis, and B. intermedius.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinobacillus↗